FN Thomson Reuters Web of Science™ VR 1.0 PT J AU Gispert, S Dutra, A Lieberman, A Friedlich, D Nussbaum, RL AF Gispert, S Dutra, A Lieberman, A Friedlich, D Nussbaum, RL TI Cloning and genomic organization of the mouse gene Slc23a1 encoding a vitamin C transporter SO DNA RESEARCH LA English DT Article DE gene structure; vitamin C; transporter; oxidative stress ID ASCORBIC-ACID; BIOCHEMICAL FUNCTIONS; DEHYDROASCORBIC ACID; FAMILY; HYBRIDIZATION; SEQUENCES; INSIGHTS; CELLS AB Vitamin C is known to exist in particularly high concentrations in brain tissue, and its free radical scavenging function is thought to represent a major antioxidative defense system. We have cloned, sequenced and analyzed the genomic structure of a mouse sodium-dependent vitamin C transporter gene, Slc23a1 (also known as Svct2). The mouse Slc23a1 cDNA is 6.4 kb long and was cloned directly from a mouse brain RNA preparation. Hybridization screening of a mouse genomic BAC library identified BAC 53L21 which contains at least the entire coding sequence of the mouse Slc23a1 gene. Determination of the exon-intron structure of the gene revealed 17 exons ranging from 58 bp to 4407 bp extending over 50 kb of the mouse genome, with the translation start codon located in exon 3. Its 1944 nucleotide open reading frame encodes a polypeptide of 647 aa, which is highly similar to rat and human orthologs. The mouse gene was assigned to chromosome 2qG2 by fluorescence in situ hybridization analysis. Expression of this gene was demonstrated in a wide range of tissues, with especially high levels in brain. Neurodegenerative diseases with an established role for oxidative stress in the cytoplasm may therefore be conditions of SLC23A1 dysfunction. C1 NHGRI, Genet Dis Res Branch, NIH, Bethesda, MD 20892 USA. Natl Inst Neurol Dis & Stroke, Neurogenet Branch, Bethesda, MD USA. NIH, Howard Hughes Med Inst, Bethesda, MD USA. RP Nussbaum, RL (reprint author), NHGRI, Genet Dis Res Branch, NIH, Bldg 49,Room 4A72,49 Convent Dr, Bethesda, MD 20892 USA. NR 22 TC 9 Z9 11 U1 0 U2 8 PU UNIVERSAL ACADEMY PRESS INC PI TOKYO PA ORDER DEPT., C P O BOX 235, TOKYO, 100-8691, JAPAN SN 1340-2838 J9 DNA RES JI DNA Res. PD DEC 31 PY 2000 VL 7 IS 6 BP 339 EP 345 DI 10.1093/dnares/7.6.339 PG 7 WC Genetics & Heredity SC Genetics & Heredity GA 403EM UT WOS:000167029700004 PM 11214969 ER PT J AU Spirina, O Bykhovskaya, Y Kajava, AV O'Brien, TW Nierlich, DP Mougey, EB Sylvester, JE Graack, HR Wittmann-Liebold, B Fischel-Ghodsian, N AF Spirina, O Bykhovskaya, Y Kajava, AV O'Brien, TW Nierlich, DP Mougey, EB Sylvester, JE Graack, HR Wittmann-Liebold, B Fischel-Ghodsian, N TI Heart-specific splice-variant of a human mitochondrial ribosomal protein (mRNA processing; tissue specific splicing) SO GENE LA English DT Article DE heart-specific splice-variant; human mitochondrial ribosomal protein; mRNA processing; tissue specific splicing ID DATABASE; RNA AB It has been proposed that splice-variants of proteins involved in mitochondrial RNA processing and translation may be involved in the tissue specificity of mitochondrial DNA disease mutations (Fischel-Ghodsian, 1998. Mel. Genet. Metab. 65, 97-104). To identify and characterize the structural components of mitochondrial RNA processing and translation, the Mammalian Mitochondrial Ribosomal Consortium has been formed. The 338 amino acid (aa) residues long MRP-LS was identified (O'Brien et al., 1999. J. Biol. Chem. 274, 36043-36051), and its transcript was screened for tissue specific splice-variants. Screening of the EST databases revealed a single putative splice-variant, due to the insertion of an exon consisting of 89 nucleotides prior to the last exon. Screening of multiple cDNA libraries revealed this inserted exon to be present only in heart tissue, in addition to the predominant MRP-LS transcript. Sequencing of this region confirmed the EST sequence, and showed in the splice-variant a termination triplet at the beginning of the last exon. Thus the inserted exon replaces the coding sequence of the regular last exon, and creates a new 353 aa long protein (MRP-L5V1). Sequence analysis and 3D modeling reveal similarity between MRP-LS and threonyl-t-RNA synthetases, and a likely RNA binding site within MRP-LS, with the C-terminus in proximity to the RNA binding site, Sequence analysis of MRP-L5V1 also suggests a likely transmembrane domain at the C-terminus. Thus it is possible that the MRP-L5V1 C-terminus could interfere with RNA binding and may have gained a transmembrane domain. Further studies will be required to elucidate the functional significance of MRP-L5V1. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Cedars Sinai Med Ctr, Burns & Allen Res Inst, Med Genet Birth Defects Ctr,Ahmanson Dept Pediat, Steven Spielberg Pediat Res Ctr, Los Angeles, CA 90048 USA. Univ Calif Los Angeles, Sch Med, Los Angeles, CA USA. NIH, CIT, Ctr Mol Modeling, Bethesda, MD USA. Univ Florida, Gainesville, FL USA. Univ Calif Los Angeles, Los Angeles, CA USA. Nemours Childrens Clin, Jacksonville, FL USA. Max Delbruck Ctr Mol Med, Berlin, Germany. RP Fischel-Ghodsian, N (reprint author), Cedars Sinai Med Ctr, Dept Pediat, WT 1165,8700 Beverly Blvd, Los Angeles, CA 90048 USA. RI Kajava, Andrey/E-1107-2014 OI Kajava, Andrey/0000-0002-2342-6886 FU NIDCD NIH HHS [R01DC04092] NR 22 TC 15 Z9 16 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-1119 J9 GENE JI Gene PD DEC 31 PY 2000 VL 261 IS 2 BP 229 EP 234 DI 10.1016/S0378-1119(00)00504-7 PG 6 WC Genetics & Heredity SC Genetics & Heredity GA 401YK UT WOS:000166956600005 PM 11167009 ER PT J AU Ikonomi, P Noguchi, CT Miller, W Kassahun, H Hardison, R Schechter, AN AF Ikonomi, P Noguchi, CT Miller, W Kassahun, H Hardison, R Schechter, AN TI Levels of GATA-1/GATA-2 transcription factors modulate expression of embryonic and fetal hemoglobins SO GENE LA English DT Article DE transcriptional factors; globin expression; hematopoietic erythroid cells; GATA-binding sites ID EPSILON-GLOBIN GENE; FACTOR GATA-1; HEMATOPOIETIC-CELLS; REGULATORY ELEMENTS; CHROMATIN STRUCTURE; FINGER DOMAINS; DIFFERENTIATION; PROTEIN; ENHANCER; PROMOTER AB GATA transcription factors bind the consensus sequence WGATAR, present in the flanking regions of most erythroid specific genes. GATA-1 and GATA-2, coexpressed in erythroid cells, are important for expression of erythroid genes. To elucidate the role of specific GATA transcription factors on globin gene expression, we examined the human alpha- and beta -globin gene clusters for all GATA sites. Conserved GATA sites were found in each of the hypersensitive sites in both beta -and alpha clusters and in proximal regulatory regions of the zeta-, epsilon- and gamma -globin but not the alpha, delta or beta -globin genes. We then tested the effect of increasing levels of GATA-1 and GATA-2 on the expression of endogenous globin genes in human erythroid cells. Increasing GATA-1 levels in K562 cells decreased the levels of epsilon -globin mRNA but had no effect on the levels of expression of gamma, zeta or alpha -globin genes. Increasing GATA-2 levels increased epsilon -globin and gamma -globin transcripts. Increasing levels of GATA-1 also caused a decrease in the expression of endogenous GATA-2, while increased levels of GATA-2 had no effect on GATA-1 mRNA. Our results indicate a differential role of GATA-1 and -2 transcription factors on globin transcripts and suggest a correlation between the conservation of GATA sites in the regulatory regions and the ability of endogenous globin genes to respond to GATA transcription factors. They also suggest that quantitative changes in the levels of GATA-1 or GATA-2 can result in alterations of globin target gene expression and may participate in the ontogenic control of the globin genes. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIDDKD, Biol Chem Lab, NIH, Bethesda, MD 20892 USA. Penn State Univ, Dept Comp Sci & Engn, University Pk, PA 16802 USA. Penn State Univ, Ctr Gene Regulat, Dept Biochem & Mol Biol, University Pk, PA 16802 USA. RP Schechter, AN (reprint author), NIDDKD, Biol Chem Lab, NIH, Bldg 10,Room 9N-307,10 Ctr Dr,MSC 1822, Bethesda, MD 20892 USA. RI Hardison, Ross/G-1142-2010; OI Hardison, Ross/0000-0003-4084-7516; Schechter, Alan N/0000-0002-5235-9408 NR 33 TC 42 Z9 45 U1 2 U2 7 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-1119 J9 GENE JI Gene PD DEC 31 PY 2000 VL 261 IS 2 BP 277 EP 287 DI 10.1016/S0378-1119(00)00510-2 PG 11 WC Genetics & Heredity SC Genetics & Heredity GA 401YK UT WOS:000166956600011 PM 11167015 ER PT J AU Kino, T Kopp, JB Chrousos, GP AF Kino, T Kopp, JB Chrousos, GP TI Glucocorticoids suppress human immunodeficiency virus type-1 long terminal repeat activity in a cell type-specific, glucocorticoid receptor-mediated fashion: direct protective effects at variance with clinical phenomenology SO JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY LA English DT Article DE glucocorticoid receptor; glucocorticoids; human immunodeficiency virus; protective effects; type-1 long terminal repeat activity ID STEROID-HORMONE RECEPTORS; SP1 TRANSCRIPTION FACTOR; HIV GENE-EXPRESSION; NF-KAPPA-B; CROSS-TALK; IN-VITRO; ADRENAL-STEROIDS; INFECTED MEN; TAT PROTEIN; PROMOTER AB Glucocorticoid administration and/or excess secretion have been associated with increased Human Immunodeficiency Virus Type-1 (HIV-1) replication and AIDS progression. The HIV-1 long terminal repeat (LTR) promoter contains glucocorticoid-responsive element (GRE)-like sequences that could mediate a positive effect of glucocorticoids on HIV-1. In addition, we recently demonstrated that the HIV-1 accessory protein Vpr is a potent coactivator of the glucocorticoid receptor. which, like the host coactivator p300, potentiates the effect of glucocorticoids on GRE-containing. glucocorticoid-responsive genes. Such an effect may increase the sensitivity of several host target tissues to glucocorticoids by several fold, and may, thus, contribute to a positive effect of glucocorticoids on the HIV-1-LTR in infected host cells. In this study, we determined the direct effect of glucocorticoids on HIV-1-LTR by examining the ability of dexamethasone to modulate the activity of this promoter coupled to the luciferase reporter gene in human cell lines. Dexamethasone markedly inhibited Tat-stimulated, p300- or Vpr-enhanced luciferase activities in a cell-type specific, dose-dependent. and glucocorticoid receptor-mediated fashion. This effect of dexamethasone was not potentiated by Vpr, was antagonized by the glucocorticoid receptor antagonist RU 486 and required the DNA-binding domain of the receptor. These data suggest that the inhibitory effect of glucocorticoids on the HIV-1-LTR may be exerted via non-GRE-dependent inhibition of the strongly positive host transcription factor NF-kappaB, which interacts with the DNA- and ligand-binding domains of the receptor. Alternatively, it is also possible that dexamethasone-activated glucocorticoid receptor competes with other transcription factors for their binding sites on the promoter region or squelches transcription factors shared by HIV-1-LTR and glucocorticoid-responsive promoters. We conclude that glucocorticoids suppress, rather than stimulate, the HIV-1 promoter, thus acting, protectively for the host. Their apparent negative clinical association with AIDS is most likely due to immunosuppression of the host. Published by Elsevier Science Ltd. C1 NICHHD, Pediat & Reprod Endocrinol Branch, NIH, Bethesda, MD 20892 USA. NIDDKD, Kidney Dis Sect, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. RP Kino, T (reprint author), NICHHD, Pediat & Reprod Endocrinol Branch, NIH, Bldg 10,Room 9D42,10 Ctr Dr,MSC 1583, Bethesda, MD 20892 USA. OI Kopp, Jeffrey/0000-0001-9052-186X NR 59 TC 24 Z9 24 U1 0 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0960-0760 J9 J STEROID BIOCHEM JI J. Steroid Biochem. Mol. Biol. PD DEC 31 PY 2000 VL 75 IS 4-5 BP 283 EP 290 DI 10.1016/S0960-0760(00)00187-4 PG 8 WC Biochemistry & Molecular Biology; Endocrinology & Metabolism SC Biochemistry & Molecular Biology; Endocrinology & Metabolism GA 412RD UT WOS:000167567400012 PM 11282284 ER PT J AU Zhao, YG Hermesz, E Yarolin, MC Westphal, H AF Zhao, YG Hermesz, E Yarolin, MC Westphal, H TI Genomic structure, chromosomal localization and expression of the human LIM-homeobox gene LHX5 SO GENE LA English DT Article DE central nervous system; chromosome 12; gene expression; homeobox ID LIM/HOMEOBOX GENE; HOMEODOMAIN PROTEINS; SEQUENCE; DIFFERENTIATION; INITIATOR; SUGGESTS; CLONING; LINKAGE; SITES AB The LIM-homeobox gene Lhx5 plays an essential role in the regulation of neuronal differentiation and migration during development of the central nervous system. Mice lacking Lhx5 function show severely disorganized brain morphology and are impaired in cognition and motor coordination. In this study, we characterized the cDNA and genomic organization of the human LHX5 gene and analyzed its expression and chromosomal location. The human gene was found to contain five exons encoding a protein composed of 402 amino acids that is 98.8% identical to mouse Lhx5. By reverse transcriptase polymerase chain reaction, LHX5 transcripts were detected in fetal brain and in various regions of the adult central nervous system including the spinal cord, the thalamus, and the cerebellum. Fluorescence in situ hybridization mapped the LHX5 gene to chromosome 12, position 12q24.31-24.32. These results provide a framework for future analysis of possible association of human hereditary disorders with mutations in LHX5. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NICHHD, Lab Mammalian Genes & Dev, NIH, Bethesda, MD 20892 USA. Univ Szeged, Fac Sci, Dept Biochem, H-6701 Szeged, Hungary. RP Westphal, H (reprint author), NICHHD, Lab Mammalian Genes & Dev, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. EM hw@helix.nih.gov NR 27 TC 8 Z9 9 U1 0 U2 3 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-1119 EI 1879-0038 J9 GENE JI Gene PD DEC 30 PY 2000 VL 260 IS 1-2 BP 95 EP 101 DI 10.1016/S0378-1119(00)00466-2 PG 7 WC Genetics & Heredity SC Genetics & Heredity GA 387UX UT WOS:000166141200010 PM 11137295 ER PT J AU Andreola, F Giandomenico, V Spero, R De Luca, LM AF Andreola, F Giandomenico, V Spero, R De Luca, LM TI Expression of a smaller lecithin : retinol acyl transferase transcript and reduced retinol esterification in MCF-7 cells SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE LRAT; RAR; breast cancer; retinyl esters ID ACID; CARCINOMA; METABOLISM; LINES AB Retinyl ester concentration is regulated by retinoic acid (RA) through an autoregulatory loop, which acts on lecithin:retinol acyltransferase (LRAT). me tested whether retinol esterification activity is downregulated in human mammary carcinoma cells and whether LRAT expression is RAR-regulated. Normal human mammary epithelial (HMEC) cells expressed a retinoid-upregulated 5-kb LRAT transcript and synthesized retinyl esters from H-3-retinol. Human carcinoma MCF-7 cells failed to express the 5-kb LRAT transcript and to synthesize retinyl esters. Instead, they expressed a 2.7-kb LRAT transcript. Both transcripts were upregulated by RA. Stable expression of the dominant-negative RAR alpha 403 blunted the up-regulation of LRAT mRNA by RA, We conclude that retinol esterification is decreased in MCF-7 vs normal mammary cells; that these cancer cells express a shorter (2.7 kb) LRAT transcript, and that retinoid receptors are involved in the regulation of LRAT-mediated retinyl ester synthesis by RA. (C) 2000 Academic Press. C1 NCI, Cellular Carcinogenesis & Tumor Promot Lab, Bethesda, MD 20892 USA. RP De Luca, LM (reprint author), NIH Bldg 37,Room 3A-17,37 Convent Dr, Bethesda, MD 20892 USA. NR 15 TC 17 Z9 17 U1 0 U2 2 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD DEC 29 PY 2000 VL 279 IS 3 BP 920 EP 924 DI 10.1006/bbrc.2000.3995 PG 5 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 391ZJ UT WOS:000166385900029 PM 11162450 ER PT J AU Lopaczynski, W Terry, C Nissley, P AF Lopaczynski, W Terry, C Nissley, P TI Autophosphorylation of the insulin-like growth factor I receptor cytoplasmic domain SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE 6-His tag; insulin receptor; IGF-I receptor dimer ID BACULOVIRUS EXPRESSION SYSTEM; PROTEIN-TYROSINE KINASE; SERINE KINASE; CIS-AUTOPHOSPHORYLATION; HYBRID RECEPTORS; PURIFICATION; VIRUS AB The cytoplasmic domain of the beta subunit of the insulin-like growth factor I receptor (amino acids 936-1337) was overexpressed in Sf9 insect cells using a baculovirus expression system, and the g-His tagged receptor was purified by metal-affinity chromatography. Autophosphorylation of the receptor was concentration dependent, consistent with a trans phosphorylation mechanism, Phosphoamino acid analysis of the autophosphorylated receptor showed predominantly phosphotyrosine, but phosphoserine and phosphothreonine were also present. However, when the receptor was further purified by gel filtration on Sephadex G-100 and then autophosphorylated, phosphoaminoacid analysis showed only phosphotyrosine, We conclude that the IGF-I receptor tyrosine kinase is not a dual-specificity kinase and that autophosphorylation of the beta subunit is by a trans mechanism. C1 NCI, Metab Branch, NIH, Bethesda, MD 20892 USA. RP Lopaczynski, W (reprint author), NCI, Metab Branch, NIH, Bethesda, MD 20892 USA. NR 25 TC 19 Z9 20 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD DEC 29 PY 2000 VL 279 IS 3 BP 955 EP 960 DI 10.1006/bbrc.2000.4046 PG 6 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 391ZJ UT WOS:000166385900035 PM 11162456 ER PT J AU Hong, F Nguyen, VA Shen, XN Kunos, G Gao, B AF Hong, F Nguyen, VA Shen, XN Kunos, G Gao, B TI Rapid activation of protein kinase B/Akt has a key role in antiapoptotic signaling during liver regeneration SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE PKB/Akt; PI3 kinase; Bad; liver regeneration; apoptosis; growth factors ID GROWTH-FACTOR-BETA; TUMOR-NECROSIS-FACTOR; NF-KAPPA-B; RAT-LIVER; TRANSFORMING GROWTH-FACTOR-BETA-1; MESSENGER-RNA; BCL-X; APOPTOSIS; HEPATOCYTES; TRANSCRIPTION AB Liver regeneration is controlled by multiple signaling pathways induced by a variety of growth factors, hormones, and cytokines. Here we report that protein kinase B (PKB)/Akt, part of a key cell survival signaling pathway, is markedly activated after partial hepatectomy (PHX). The antiapoptotic protein Bad, a downstream target of PKB/Akt, is also phosphorylated, This cascade can be activated by various factors in primary hepatocytes, with the strongest activation by insulin and the alpha (1)-adrenergic agonist phenylephrine (PE), followed by IL-6, epidermal growth factor (EGF), and hepatocyte growth factor (HGF). Pretreatment of cells with the specific PI3 kinase inhibitor LY294002 abolished insulin- or PE-activation of PKB/ Akt, suggesting that activation of PKB/Akt is mediated by a PI3 kinase-dependent mechanism. In vivo administration of PE, insulin, IL-6, HGF, or EGF to mice markedly stimulated PKB/Akt in the liver, with the strongest stimulation induced by insulin and PE. Moreover, HGF and insulin were able to attenuate transforming growth factor beta -induced apoptosis in hepatic cells, and these effects were antagonized by LY294002. Taken together, these Findings suggest that rapid activation of PKB/Akt is a key antiapoptotic signaling pathway involved in liver regeneration. (C) 2000 Academic Press. C1 NIAAA, Sect Liver Biol, NIH, Bethesda, MD 20892 USA. NIAAA, Lab Physiol Studies, NIH, Bethesda, MD 20892 USA. Virginia Commonwealth Univ, Med Coll Virginia, Dept Pharmacol & Toxicol, Richmond, VA 23298 USA. RP Gao, B (reprint author), NIAAA, Sect Liver Biol, NIH, Bethesda, MD 20892 USA. NR 41 TC 54 Z9 56 U1 1 U2 7 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD DEC 29 PY 2000 VL 279 IS 3 BP 974 EP 979 DI 10.1006/bbrc.2000.4044 PG 6 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 391ZJ UT WOS:000166385900039 PM 11162460 ER PT J AU Romanin, C Gamsjaeger, R Kahr, H Schaufler, D Carlson, O Abernethy, DR Soldatov, NM AF Romanin, C Gamsjaeger, R Kahr, H Schaufler, D Carlson, O Abernethy, DR Soldatov, NM TI Ca2+ sensors of L-type Ca2+ channel SO FEBS LETTERS LA English DT Article DE heart; calcium channel; calmodulin; Ca2+ sensor; inactivation ID CA2+-DEPENDENT INACTIVATION; ALPHA(1C) SUBUNIT; CALCIUM CHANNELS; CALMODULIN; INHIBITION; DEPENDS; MOTIF AB Ca2+-induced inactivation of L-type Ca2+ is differentially mediated by two C-terminal motifs of the ale subunit, L (1572-1587) and K (1599-1651) implicated for calmodulin binding. We found that motif L is composed of a highly selective Ca2+ sensor and an adjacent Ca2+-independent tethering site for calmodulin, The Ca sensor contributes to higher Ca2+ sensitivity of the motif L complex with calmodulin. Since only combined mutation of both sites removes Ca2+-dependent current decay, the two-site modulation by Ca2+ and calmodulin may underlie Ca2+-induced inactivation of the channel. (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V, All rights reserved.Ca2+-induced inactivation of L-type Ca2+ is differentially mediated by two C-terminal motifs of the ale subunit, L (1572-1587) and K (1599-1651) implicated for calmodulin binding. We found that motif L is composed of a highly selective Ca2+ sensor and an adjacent Ca2+-independent tethering site for calmodulin, The Ca sensor contributes to higher Ca2+ sensitivity of the motif L complex with calmodulin. Since only combined mutation of both sites removes Ca2+-dependent current decay, the two-site modulation by Ca2+ and calmodulin may underlie Ca2+-induced inactivation of the channel. (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved. C1 NIA, NIH, Baltimore, MD 21224 USA. Univ Linz, Inst Biophys, A-4040 Linz, Austria. RP Soldatov, NM (reprint author), NIA, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Romanin, Christoph/D-5399-2009; OI Romanin, Christoph/0000-0003-3756-4136; Gamsjaeger, Roland/0000-0003-1095-2569 NR 21 TC 64 Z9 64 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0014-5793 J9 FEBS LETT JI FEBS Lett. PD DEC 29 PY 2000 VL 487 IS 2 BP 301 EP 306 DI 10.1016/S0014-5793(00)02361-9 PG 6 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 392AL UT WOS:000166388400032 PM 11150529 ER PT J AU Bandyopadhyay, G Sajan, MP Kanoh, Y Standaert, ML Burke, TR Quon, MJ Reed, BC Dikic, I Noel, LE Newgard, CB Farese, R AF Bandyopadhyay, G Sajan, MP Kanoh, Y Standaert, ML Burke, TR Quon, MJ Reed, BC Dikic, I Noel, LE Newgard, CB Farese, R TI Glucose activates mitogen-activated protein kinase (extracellular signal-regulated kinase) through proline-rich tyrosine kinase-2 and the glut1 glucose transporter SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID INSULIN-STIMULATED TRANSLOCATION; RAT ADIPOSE CELL; C-ZETA; MAP KINASE; INCREASES DIACYLGLYCEROL; ADIPOCYTES; RECEPTOR; BETA; SECRETION; ALPHA AB Glucose serves as both a nutrient and regulator of physiological and pathological processes. Presently, we found that glucose and certain sugars rapidly activated extracellular signal-regulated kinase (ERK) by a mechanism that was: (a) independent of glucose uptake/metabolism and protein kinase C but nevertheless cytochalasin B-inhibitable; (b) dependent upon proline-rich tyrosine kinase-2 (PYK2), GRBB, SOS, RAS, RAF, and MEK1; and (c) amplified by overexpression of the Glut1, but not Glut2, Gluts, or Glut4, glucose transporter. This amplifying effect was independent of glucose uptake but dependent on residues 463-468, IASGFR, in the Glut1 C terminus. Accordingly, glucose effects on ERK were amplified by expression of Glut 4/Glut1 or Glut2/Glut1 chimeras containing IASGFR but not by Glut1/Glut4 or Glut1/Grlut4 chimeras lacking these residues. Also, deletion of Glut1 residues 469-492 was without effect, but mutations involving serine 465 or arginine 468 yielded dominant-negative forms that inhibited glucose-dependent ERK activation. Glucose stimulated the phosphorylation of tyrosine residues 402 and 881 in PYK2 and binding of PYK2 to Myc-Glut1. Our findings suggest that: (a) glucose activates the GRB2/SOS/RAS/RAF/ MEK1/ERK pathway by a mechanism that requires PYK2 and residues 463-468, LASGFR, in the Glut1 C terminus and (b) Glutl serves as a sensor, transducer, and amplifier for glucose signaling to PYK2 and ERK. C1 James A Haley Vet Hosp, Res Serv VAR 151, Tampa, FL 33612 USA. Univ S Florida, Coll Med, Dept Internal Med, Tampa, FL 33612 USA. NHLBI, Hypertens Endocrine Branch, NIH, Bethesda, MD 20892 USA. NCI, NIH, Bethesda, MD 20892 USA. Louisiana State Univ, Hlth Sci Ctr, Dept Biochem & Mol Biol, Shreveport, LA 71130 USA. Ludwig Inst Canc Res, S-75124 Uppsala, Sweden. Univ Texas, SW Med Ctr, Dept Biochem, Dallas, TX 75235 USA. Univ Texas, SW Med Ctr, Dept Internal Med, Dallas, TX 75235 USA. Univ Texas, SW Med Ctr, Gifford Labs Diabet Res, Dallas, TX 75235 USA. RP Farese, R (reprint author), James A Haley Vet Hosp, Res Serv VAR 151, 13000 Bruce B Downs Blvd, Tampa, FL 33612 USA. RI Quon, Michael/B-1970-2008; Burke, Terrence/N-2601-2014; Dikic, Ivan/O-4650-2015; OI Dikic, Ivan/0000-0001-8156-9511; Quon, Michael/0000-0002-9601-9915; Quon , Michael /0000-0002-5289-3707 FU NIDDK NIH HHS [2R01DK38079-09A1] NR 40 TC 50 Z9 51 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 29 PY 2000 VL 275 IS 52 BP 40817 EP 40826 DI 10.1074/jbc.M007920200 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 387GJ UT WOS:000166114600023 PM 11007796 ER PT J AU Piatigorsky, J Kozmik, Z Horwitz, J Ding, LL Carosa, E Robison, WG Steinbach, PJ Tamm, ER AF Piatigorsky, J Kozmik, Z Horwitz, J Ding, LL Carosa, E Robison, WG Steinbach, PJ Tamm, ER TI Omega-crystallin of the scallop lens - A dimeric aldehyde dehydrogenase class 1/2 enzyme-crystallin SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ALPHA-B-CRYSTALLIN; SITE-DIRECTED MUTAGENESIS; HORMONE-BINDING PROTEIN; MAJOR SOLUBLE-PROTEIN; HEAT-SHOCK PROTEIN; EYE LENS; CORNEAL EPITHELIUM; STRUCTURAL PROTEIN; BOVINE CORNEA; MOUSE CORNEA AB While many of the diverse crystallins of the transparent lens of vertebrates are related or identical to metabolic enzymes, much less is known about the lens crystallins of invertebrates. Here we investigate the complex eye of scallops. Electron microscopic inspection revealed that the anterior, single layered corneal epithelium overlying the cellular lens contains a regular array of microvilli that we propose might contribute to its optical properties. The sole crystallin of the scallop eye lens was found to be homologous to Omega -crystallin, a minor crystallin in cephalopods related to aldehyde dehydrogenase (ALDH) class 1/2. Scallop Omega -crystallin (officially designated ALDH1A9) is 55-56% identical to its cephalopod homologues, while it is 67 and 64% identical to human ALDH 2 and 1, respectively, and 61% identical to retinaldehyde dehydrogenase/eta -crystallin of elephant shrews.:like other enzyme-crystallins, scallop Omega -crystallin appears to be present in low amounts in nonocular tissues. Within the scallop eye, immunofluorescence tests indicated that Omega -crystallin expression is confined to the lens and cornea, Although it has conserved the critical residues required for activity in other ALDHs and appears by homology modeling to have a structure very similar to human ALDH2, scallop Omega -crystallin was enzymatically inactive with diverse substrates and did not bind NAD or NADP, In contrast to mammalian ALDH1 and -2 and other cephalopod Omega -crystallins, which are tetrameric proteins, scallop Omega -crystallin is a dimeric protein. Thus, ALDH is the most diverse lens enzyme-crystallin identified so far, having been used as-a lens crystallin in at least two classes of molluscs as well as elephant shrews. C1 NEI, Mol & Dev Biol Lab, NIH, Bethesda, MD 20892 USA. NEI, Lab Mechanisms Ocular Dis, NIH, Bethesda, MD 20892 USA. NIH, Ctr Mol Modeling, Ctr Informat Technol, Bethesda, MD 20892 USA. Acad Sci Czech Republ, Inst Mol Genet, Lab Transcript Regulat, Prague 6, Czech Republic. Univ Calif Los Angeles, Sch Med, Jules Stein Eye Inst, Los Angeles, CA 90095 USA. RP Piatigorsky, J (reprint author), NEI, Mol & Dev Biol Lab, NIH, Bldg 6,Rm 201, Bethesda, MD 20892 USA. RI Kozmik, Zbynek/G-3581-2014; Kozmik, Zbynek/I-8807-2014 NR 79 TC 32 Z9 32 U1 0 U2 6 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 29 PY 2000 VL 275 IS 52 BP 41064 EP 41073 DI 10.1074/jbc.M005625200 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 387GJ UT WOS:000166114600055 PM 10961997 ER PT J AU Prufer, K Racz, A Lin, GC Barsony, J AF Prufer, K Racz, A Lin, GC Barsony, J TI Dimerization with retinoid X receptors promotes nuclear localization and subnuclear targeting of vitamin D receptors SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID FLUORESCENT PROTEIN CHIMERA; LIVING CELLS; PROGESTERONE-RECEPTOR; 1,25-DIHYDROXYVITAMIN D-3; ACID RECEPTORS; DNA-BINDING; IN-VIVO; EXPRESSION; VISUALIZATION; TRANSCRIPTION AB The vitamin D receptor (VDR) acts as heterodimer with the retinoid X receptor alpha (RXR) to control transcriptional activity of target genes. To explore the influence of heterodimerization on the subcellular distribution of these receptors in living cells, we developed a series of fluorescent-protein chimeras. The steady-state distribution of the yellow fluorescent protein-RXR was more nuclear than the unliganded green fluorescent protein (GFP)-VDR. Coexpression of RXR-blue fluorescent protein (BFP) promoted nuclear accumulation of GFP-VDR by influencing both nuclear import and retention. Fluorescence resonance energy transfer microscopy (FRET) demonstrated that the unliganded GFP-VDR and RXR-BFP form heterodimers. The increase in nuclear heterodimer content correlated with an increase in basal transcriptional activity. FRET also revealed that calcitriol induces formation of multiple nuclear foci of heterodimers. Mutational analysis showed a correlation; between hormone-dependent nuclear VDR foci formation and DNA binding. RXR-BFP also promoted hormone-dependent nuclear accumulation and intranuclear foci formation of a nuclear localization signal mutant receptor (nlsGFP-VDR) and rescued its transcriptional activity. Heterodimerization mutant RXR failed to alter GFP-VDR and nlsGFP-VDR distribution or activity. These experiments suggest that RXR has a profound effect on VDR distribution. This effect of RXR to promote nuclear accumulation and intranuclear targeting contributes to the regulation of VDR activity and probably the activity of other heterodimerization partners. C1 NIDDK, Lab Cell Biochem & Biol, NIH, Bethesda, MD 20892 USA. RP Barsony, J (reprint author), NIDDK, Lab Cell Biochem & Biol, NIH, Bldg 8,Rm 422,8 Ctr Dr, Bethesda, MD 20892 USA. NR 48 TC 93 Z9 95 U1 2 U2 9 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 29 PY 2000 VL 275 IS 52 BP 41114 EP 41123 DI 10.1074/jbc.M003791200 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 387GJ UT WOS:000166114600062 PM 11001945 ER PT J AU Pece, S Gutkind, JS AF Pece, S Gutkind, JS TI Signaling from E-cadherins to the MAPK pathway by the recruitment and activation of epidermal growth factor receptors upon cell-cell contact formation SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID CATENIN COMPLEX; PROTEIN INTERACTIONS; ADHESION; JUNCTIONS; INTEGRINS; SURVIVAL; LINE AB E-cadherins are well characterized cell surface molecules expressed in epithelial cells, which play a major role in cell adhesion through the establishment of calcium-dependent hemophilic interactions at sites of cell-cell contacts. They are also integral components of morphogenetic programs controlling the maintenance of the structural and functional integrity of epithelia, Accumulated evidence indicates that the E-cadherin-mediated cell adhesion system is highly regulated from inside the cells by a number of intracellular signaling pathways. Recently available information suggests that E-cadherins may also play a role in the transduction of signals from the outside of the cell to the cytoplasm. However, the nature of the biochemical routes regulated by E-cadherins is still largely unknown. In this study, we set out to explore the possibility that E-cadherins may regulate the activity of MAPK, a key signaling pathway involved in cell fate decisions, upon the formation of cell-cell contacts among neighboring cells. By using an immortalized non-tumorigenic keratinocyte cell line, HaCat, as a model system, we provide evidence that the assembly of calcium-dependent adherens junctions leads to a rapid and remarkable increase in the state of activation of MAPK and that this event is mediated by E-cadherins. Furthermore, we found that E-cadherins stimulate the MAPK pathway through the ligand-independent activation of epidermal growth factor receptors and the consequent activation of a biochemical route leading to the stimulation of MAPKs. These findings suggest that E-cadherins can initiate outside-in signal transducing pathways through the engagement of tyrosine kinase receptors for epidermal growth factor, thus providing a novel molecular mechanism whereby these cell adhesion molecules may ultimately control the fate of normal and transformed epithelial cells. C1 NIDCR, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD 20892 USA. RP Gutkind, JS (reprint author), NIDCR, Oral & Pharyngeal Canc Branch, NIH, Bldg 30,Rm 211,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Gutkind, J. Silvio/A-1053-2009; Pece, Salvatore/B-9609-2013 OI Pece, Salvatore/0000-0003-1764-3929 NR 26 TC 230 Z9 234 U1 1 U2 7 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 29 PY 2000 VL 275 IS 52 BP 41227 EP 41233 DI 10.1074/jbc.M006578200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 387GJ UT WOS:000166114600076 PM 10969083 ER PT J AU Michel, P Rassat, A Daly, JW Spande, TF AF Michel, P Rassat, A Daly, JW Spande, TF TI A stereospecific synthesis of (+/-)-5,8-disubstituted indolizidines and (+/-)-1,4-disubstituted quinolizidines found in poison frog skins SO JOURNAL OF ORGANIC CHEMISTRY LA English DT Article ID N-ACYLDIHYDROPYRIDONES; ALKALOIDS; MANTELLA AB An efficient, high-yield stereospecific route to three (+/-)-5,8-disubstituted indolizidines, (209B (I), 209I (II), 223J (III)) and two (+/-)-1,4-disubstituted quinolizidines (207I (IV), 233A (V)), racemates of alkaloids found in the skins of neotropical and Madagascan poison frogs is reported. The structures of the natural alkaloids were thereby established by chiral GC comparison with the exception of indolizidine 209B (I) for which a natural 209B could no longer be detected. C1 Ecole Normale Super, CNRS, UMR 8640, Dept Chim, F-75231 Paris 05, France. NIDDKD, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. RP Rassat, A (reprint author), Ecole Normale Super, CNRS, UMR 8640, Dept Chim, 24 Rue Lhomond, F-75231 Paris 05, France. NR 10 TC 39 Z9 39 U1 0 U2 2 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0022-3263 J9 J ORG CHEM JI J. Org. Chem. PD DEC 29 PY 2000 VL 65 IS 26 BP 8908 EP 8918 DI 10.1021/jo000666b PG 11 WC Chemistry, Organic SC Chemistry GA 386XF UT WOS:000166089600010 PM 11149832 ER PT J AU Froimowitz, M Wu, KM Moussa, A Haidar, RM Jurayj, J George, C Gardner, EL AF Froimowitz, M Wu, KM Moussa, A Haidar, RM Jurayj, J George, C Gardner, EL TI Slow-onset, long-duration 3-(3 ',4 '-dichlorophenyl)-1-indanamine monoamine reuptake blockers as potential medications to treat cocaine abuse SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID DOPAMINE UPTAKE INHIBITORS; CONFORMATIONAL-ANALYSIS; ANTIDEPRESSANT ACTIVITY; DIFFERENTIAL BINDING; RECOGNITION SITES; SQUIRREL-MONKEYS; RHESUS-MONKEYS; ANALOGS; METHYLPHENIDATE; TRANSPORTER AB A series of 3-(3',4'-dichlorophenyl)-1-indanamine monoamine reuptake blockers have been synthesized in an effort to develop a compound that could be used as a maintenance therapy to treat cocaine abuse. Since the effects of cocaine on dopamine (DA) and serotonin (5HT) transporters are important components of its pharmacological activity; the focus was;on nonselective inhibitors of monoamine transport. To reduce or eliminate the abuse potential of a DA reuptake blocker, the compounds were designed to,be slow-onset, long-duration prodrugs whose N-demethylated metabolites would have increased-activity over the parent compound with the ideal being a parent compound that has little or no activity.;To achieve this, pairs of compounds with different groups on the amine nitrogen and with and without an additional N-methyl group were synthesized. All of the synthesized compounds were screened for binding and reuptake at the cloned human DA, 5HT, and norepinephrine (NE) transporters. As previously found, trans isomers are nonselective blockers of DA, 5HT, and NE reuptake, cis isomers with small N-alkyl groups are selective blockers of 5HT reuptake, and tertiary amines of the trans compounds are less potent than the corresponding N-demethylated secondary amines as blockers of,DA reuptake. Larger N-alkyl groups in both the trans and cis series were found to reduce activity for the 5HT and NE transporters-with less effect at DA transporters. Selected trans compounds were also screened for locomotor activity in mice and generalization to a cocaine-like profile in rats. With intraperitoneal administration, all of the trans isomers showed a slow onset of at least 20 min and an extremely long duration of action in the locomotor assays. Several of the trans compounds also fully generalized to a cocaine-like pharmacological profile. An initial lead compound, the N,N-dimethyl analogue trans-1b was resolved into chirally pure enantiomers. Surprisingly, both enantiomers were found to have significant affinity for the DA transporter and to cause locomotor activation This is in contrast to the N-methyl compound in which only the (+)-enantiomer had significant activity. The absolute configuration of the more active enantiomer was determined by X-ray crystallography to be 3R,1S. C1 Pharm Eco Labs, Devens, MA 01432 USA. USN, Res Lab, Struct Matter Lab, Washington, DC 20375 USA. NIDA, Intramural Res Program, Baltimore, MD 21224 USA. RP Froimowitz, M (reprint author), Pharm Eco Labs, 25 Patton Rd, Devens, MA 01432 USA. FU NIDA NIH HHS [N01DA-4-8313, DA 09045, N01DA-3-8303] NR 58 TC 53 Z9 54 U1 0 U2 1 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0022-2623 J9 J MED CHEM JI J. Med. Chem. PD DEC 28 PY 2000 VL 43 IS 26 BP 4981 EP 4992 DI 10.1021/jm000201d PG 12 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 386DW UT WOS:000166045700008 PM 11150168 ER PT J AU May, EL Jacobson, AE Mattson, MV Traynor, JR Woods, JH Harris, LS Bowman, ER Aceto, MD AF May, EL Jacobson, AE Mattson, MV Traynor, JR Woods, JH Harris, LS Bowman, ER Aceto, MD TI Synthesis and in vitro and in vivo activity of (-)-(1R,5R,9R)- and (+)-(1S,5S,9S)-N-alkenyl-, -N-alkynyl-, and -N-cyanoalkyl-5,9-dimethyl-2 '-hydroxy-6,7-benzomorphan homologues SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID KAPPA-OPIOID-RECEPTOR; RHESUS-MONKEYS; LIGAND-BINDING; IN-VITRO; SIGMA; ALLYLNORMETAZOCINE; NORMETAZOCINES; CLASSIFICATION; PHENCYCLIDINE; DEPENDENCE AB Two of the synthesized (-)-(1R,5R,9R)-N-homologues (N-but-3-enyl- and N-but-3-ynyl-5,9-dimethyl-2'-hydroxy-6,7-benzomorphan (9, 13)) were found to be about 20 times more potent than morphine in the mouse tail-flick assay (ED50 = 0.05 mg/kg), and(-)-(1R,5R,9R) -N-but-2-ynyl-5,9-dimethyl-2'-hydroxy-6,7-benzomorphan ((-)-(LR,5R,SR)-N-but-2-ynylnormetazocine, 12) was about as potent as the opioid antagonist N-allylnormetazocine (AD(50) in the tail-flick vs morphine assay 0.3 mg/kg). All of the homologues examined had higher affinity for the kappa -opioid receptor than the mu -receptor except (-)-N-but-2-ynyl-normetazocine (12), which had a kappa/mu ratio = 7.8 and a delta/mu ratio = 118. The (-)-N-2-cyanoethyl . (3), -allyl (8), and -but-3-ynyl (13) analogues had good affinity (<10 nM) for -opioid receptors. Two homologues in the (+)-(1S,5S,9S)-normetazocine series, N-pent-4-enyl (24) and N-hex-5-enyl (25), were high-affinity and selective sigma (1)-ligands (K-i = 2 nM, sigma (2)/sigma (1) = 1250, and 1 nM, sigma (2)/sigma (1) 750, respectively); in contrast, N-allylnormetazocine (22) had relatively poor affinity at or, and its sigma (1)/sigma (2) ratio was <100. C1 Virginia Commonwealth Univ, Dept Pharmacol & Toxicol, Richmond, VA 23298 USA. NIDDK, Med Chem Lab, NIH, Bethesda, MD 20892 USA. Univ Michigan, Sch Med, Dept Pharmacol, Ann Arbor, MI 48109 USA. RP May, EL (reprint author), Virginia Commonwealth Univ, Dept Pharmacol & Toxicol, Med Coll Virginia Campus, Richmond, VA 23298 USA. FU NIDA NIH HHS [DA 00254, DA 5-8059] NR 33 TC 12 Z9 16 U1 1 U2 1 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0022-2623 J9 J MED CHEM JI J. Med. Chem. PD DEC 28 PY 2000 VL 43 IS 26 BP 5030 EP 5036 DI 10.1021/jm000317+ PG 7 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 386DW UT WOS:000166045700014 PM 11150174 ER PT J AU Camandola, S Poli, G Mattson, MP AF Camandola, S Poli, G Mattson, MP TI The lipid peroxidation product 4-hydroxy-2,3-nonenal inhibits constitutive and inducible activity of nuclear factor kappa B in neurons SO MOLECULAR BRAIN RESEARCH LA English DT Article DE Alzheimer's disease; AP-1; apoptosis; cerebral cortex; nuclear factor kappa B; stroke ID AMYLOID BETA-PEPTIDE; AMYOTROPHIC-LATERAL-SCLEROSIS; PROTECTS HIPPOCAMPAL-NEURONS; DNA-BINDING ACTIVITY; TRANSCRIPTION FACTOR; ALZHEIMERS-DISEASE; OXIDATIVE STRESS; ALDEHYDIC PRODUCT; GENE-EXPRESSION; ION HOMEOSTASIS AB Peroxidation of membrane lipids occurs in many different neurodegenerative conditions including stroke, and Alzheimer's and Parkinson's diseases. Recent findings suggest that lipid peroxidation can promote neuronal death by a mechanism involving production of the toxic aldehyde 4-hydroxy-2,3-nonenal (HNE), which may act by covalently modifying proteins and impairing their function. The transcription factor NF-kappaB can prevent neuronal death in experimental models of neurodegenerative disorders by inducing the expression of anti-apoptotic proteins including Bcl-2 and manganese superoxide dismutase. We now report that HNE selectively suppresses basal and inducible NF-kappaB DNA binding activity in cultured rat cortical neurons. Immunoprecipitation-immunoblot analyses using antibodies against HNE-conjugated proteins and p50 and p65 NF-kappaB subunits indicate that HNE does not directly modify NF-kappaB proteins. Moreover, HNE did not affect NF-kappaB DNA-binding activity when added directly to cytosolic extracts, suggesting that HNE inhibits an upstream component of the NF-kappaB signaling pathway. Inhibition of the survival-promoting NF-kappaB signaling pathway by HNE may contribute to neuronal death under conditions in which membrane lipid peroxidation occurs. (C) 2000 Elsevier Science BN. All rights reserved. C1 NIA, Gerontol Res Ctr 4F02, Neurosci Lab, Baltimore, MD 21224 USA. Univ Turin, SL Gonzaga Hosp, Dept Clin & Biol Sci, Orbassano, Italy. Johns Hopkins Univ, Sch Med, Dept Neurosci, Baltimore, MD 21205 USA. RP Mattson, MP (reprint author), NIA, Gerontol Res Ctr 4F02, Neurosci Lab, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012 NR 52 TC 36 Z9 37 U1 0 U2 3 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0169-328X J9 MOL BRAIN RES JI Mol. Brain Res. PD DEC 28 PY 2000 VL 85 IS 1-2 BP 53 EP 60 DI 10.1016/S0169-328X(00)00234-5 PG 8 WC Neurosciences SC Neurosciences & Neurology GA 392VE UT WOS:000166432600006 ER PT J AU Balijepalli, S Boyd, MR Ravindranath, V AF Balijepalli, S Boyd, MR Ravindranath, V TI Human brain thioltransferase: constitutive expression and localization by fluorescence in situ hybridization SO MOLECULAR BRAIN RESEARCH LA English DT Article DE thiol disulfide oxidoreductase; thioltransferase; oxidative stress; brain; glutathione; human ID RED-BLOOD-CELLS; OXIDATIVE STRESS; IMMUNOLOGICAL CHARACTERIZATION; GLUTAREDOXIN; PURIFICATION; GLUTATHIONE; MITOCHONDRIA; HALOPERIDOL; PROTEINS; CLONING AB Thioltransferase (glutaredoxin) is a member of the family of thiol-disulfide oxido-reductases that maintain the sulfhydryl homeostasis in cells by catalyzing thiol-disulfide interchange reactions. One of the major consequences of oxidative stress in brain is formation of protein-glutathione mixed disulfide (through oxidation of protein thiols) which can be reversed by thioltransferase during recovery of brain from oxidative stress. Here we have visualized the location of thioltransferase in brain regions from seven human tissues obtained at autopsy. Constitutively expressed thioltransferase activity was detectable in all human brains examined although inter-individual variations were seen. The enzyme activity was significantly higher in hippocampus and cerebellum as compared to other regions. Constitutive expression of thioltransferase mRNA was detectable by Northern blot analysis. Localization of thioltransferase mRNA by fluorescence in situ hybridization revealed its presence predominantly in neurons in the cerebral cortex, Purkinje and granule cell layers of the cerebellum granule cell layer of the dentate gyrus and in the pyramidal neurons of CAI, CA2 and CA3 subfields of hippocampus. These discrete neuronal concentrations of thioltransferase would be consistent with an essential role in modulating recovery of protein thiols from mixed disulfides formed during oxidative stress. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Natl Inst Mental Hlth & Neurosci, Dept Neurochem, Bangalore 560029, Karnataka, India. Natl Brain Res Ctr, New Delhi 110067, India. NCI, Frederick Canc Res & Dev Ctr, Lab Drug Discovery Res & Dev, Dev Therapeut Program, Frederick, MD 21702 USA. RP Ravindranath, V (reprint author), Natl Inst Mental Hlth & Neurosci, Dept Neurochem, Hosur Rd, Bangalore 560029, Karnataka, India. NR 28 TC 9 Z9 10 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0169-328X J9 MOL BRAIN RES JI Mol. Brain Res. PD DEC 28 PY 2000 VL 85 IS 1-2 BP 123 EP 132 DI 10.1016/S0169-328X(00)00206-0 PG 10 WC Neurosciences SC Neurosciences & Neurology GA 392VE UT WOS:000166432600014 ER PT J AU Altose, MD Redline, S Deitz, CD Quinlan, KJ Eichenhorn, MS Conway, WA Jentons, RL Braden, K Ketchum, M Wise, RA Permutt, S Rand, CS Daniel, M Santopietro, V Weeks, KA Scanlon, PD Patel, AM Utz, JP Williams, DE Caron, GM Micras, KS Buist, AS Johnson, LR Bortz, VJ Persons, SL Schueler, HA Bailey, WC Brooks, CM Gerald, LB Montiel, L Tashkin, DP Coulson, AH Kleerup, EC Li, VC Nides, MA Zuniga, IP Lee, YE Anthonisen, NR Manfreda, J Rempel-Rossum, SC Stoyko, JM Connett, JE Kjelsberg, MO Bollenbeck, MT Kurnow, KJ Madhok, TC Skeans, MA Voelker, HT Rogers, RM Owens, GR Vitale, FM Pusateri, ME Kanner, RE Villegas, GM Esplin, C Stayner, RS Bascom, R Landis, JR Maurer, JR Phillips, Y Rennard, SI Stoller, JK Tager, I Thomas, A Hyatt, RE Lambrew, CT Mason, BA Hurd, SS Weinmann, G Wu, MC AF Altose, MD Redline, S Deitz, CD Quinlan, KJ Eichenhorn, MS Conway, WA Jentons, RL Braden, K Ketchum, M Wise, RA Permutt, S Rand, CS Daniel, M Santopietro, V Weeks, KA Scanlon, PD Patel, AM Utz, JP Williams, DE Caron, GM Micras, KS Buist, AS Johnson, LR Bortz, VJ Persons, SL Schueler, HA Bailey, WC Brooks, CM Gerald, LB Montiel, L Tashkin, DP Coulson, AH Kleerup, EC Li, VC Nides, MA Zuniga, IP Lee, YE Anthonisen, NR Manfreda, J Rempel-Rossum, SC Stoyko, JM Connett, JE Kjelsberg, MO Bollenbeck, MT Kurnow, KJ Madhok, TC Skeans, MA Voelker, HT Rogers, RM Owens, GR Vitale, FM Pusateri, ME Kanner, RE Villegas, GM Esplin, C Stayner, RS Bascom, R Landis, JR Maurer, JR Phillips, Y Rennard, SI Stoller, JK Tager, I Thomas, A Hyatt, RE Lambrew, CT Mason, BA Hurd, SS Weinmann, G Wu, MC CA Lung Hlth Study Res Grp TI Effect of inhaled triamcinolone on the decline in pulmonary function in chronic obstructive pulmonary disease. SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Article ID LONG-TERM TREATMENT; RANDOMIZED CONTROLLED TRIAL; LUNG HEALTH; BRONCHODILATOR THERAPY; CORTICOSTEROID USE; COPD; ASTHMA; BUDESONIDE; HYPERRESPONSIVENESS; SMOKING AB Background: Chronic obstructive pulmonary disease (COPD) results from a progressive decline in lung function, which is thought to be the consequence of airway inflammation. We hypothesized that antiinflammatory therapy with inhaled corticosteroids would slow this decline. Methods: We enrolled 1116 persons with COPD whose forced expiratory volume in one second (FEV1) was 30 to 90 percent of the predicted value in a 10-center, placebo-controlled, randomized trial of inhaled triamcinolone acetonide administered at a dose of 600 microg twice daily. The primary outcome measure was the rate of decline in FEV1) after the administration of a bronchodilator. The secondary outcome measures included respiratory symptoms, use of health care services, and airway reactivity. In a substudy of 412 participants, we measured bone density in the lumbar spine and femur at base line and one and three years after the beginning of treatment. Results: The mean duration of follow-up was 40 months. The rate of decline in the FEV1) after bronchodilator use was similar in the 559 participants in the triamcinolone group and the 557 participants in the placebo group (mean [+/-SE], 44.2+/-2.9 vs. 47.0+/-3.0 ml per year, P=0.50). Members of the triamcinolone group had fewer respiratory symptoms during the course of the study (21.1 per 100 person-years vs. 28.2 per 100 person-years, P=0.005) and had fewer visits to a physician because of a respiratory illness (1.2 per 100 person-years vs. 2.1 per 100 person-years, P=0.03). Those taking triamcinolone also had lower airway reactivity in response to methacholine challenge at 9 months and 33 months (P=0.02 for both comparisons). After three years, the bone density of the lumbar spine (P=0.007) and the femur (P<0.001) was significantly lower in the triamcinolone group. Conclusions: Inhaled triamcinolone does not slow the rate of decline in lung function in people with COPD, but it improves airway reactivity and respiratory symptoms and decreases the use of health care services for respiratory problems. These benefits should be weighed against the potential long-term adverse effects of triamcinolone on bone mineral density. (N Engl J Med 2000;343:1902-9.) (C) 2000, Massachusetts Medical Society. C1 Lung Hlth Study Coordinating Ctr, Minneapolis, MN 55414 USA. Mayo Clin & Mayo Fdn, Rochester, MN 55905 USA. Oregon Hlth Sci Univ, Portland, OR 97201 USA. Univ Pittsburgh, Pittsburgh, PA 15260 USA. NHLBI, Div Epidemiol & Clin Applicat, Bethesda, MD 20205 USA. RP Connett, JE (reprint author), Lung Hlth Study Coordinating Ctr, 2221 Univ Ave SE,Suite 200, Minneapolis, MN 55414 USA. RI Landis, J. Richard/A-9330-2010; OI Wise, Robert/0000-0002-8353-2349 NR 39 TC 474 Z9 479 U1 1 U2 9 PU MASSACHUSETTS MEDICAL SOC PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD DEC 28 PY 2000 VL 343 IS 26 BP 1902 EP 1909 PG 8 WC Medicine, General & Internal SC General & Internal Medicine GA 386VR UT WOS:000166082700001 ER PT J AU Houston, AL Chen, HC Schatz, BR Hubbard, SM Sewell, RR Ng, TD AF Houston, AL Chen, HC Schatz, BR Hubbard, SM Sewell, RR Ng, TD TI Exploring the use of concept spaces to improve medical information retrieval SO DECISION SUPPORT SYSTEMS LA English DT Article DE Information retrieval; medical informatics; medical information retrieval; concept space; MeSH terms; UMLS metathesaurus ID QUERY EXPANSION; SYSTEM; DATABASES; MODEL AB This research investigated the application of techniques successfully used in previous information retrieval research, to the more challenging area of medical informatics. It was performed on a biomedical document collection testbed, CANCERLIT, provided by the National Cancer Institute (NCI), which contains information on all types of cancer therapy. The quality or usefulness of terms suggested by three different thesauri, one based on MeSH terms, one based solely on terms from the document collection, and one based on the Unified Medical Language System (UMLS) Metathesaurus, was explored with the ultimate goal of improving CANCERLIT information search and retrieval. Researchers affiliated with the University of Arizona Cancer Center evaluated lists of related terms suggested by different thesauri for 12 different directed searches in the CANCERLIT testbed. The preliminary results indicated that among the thesauri, there were no statistically significant differences in either term recall or precision. Surprisingly, there was almost no overlap of relevant terms suggested by the different thesauri for a given search. This suggests that recall could be significantly improved by using a combined thesaurus approach. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Louisiana State Univ, Coll Business Adm, ISDS Dept, Baton Rouge, LA 70803 USA. Univ Arizona, Dept Management Informat Syst, Tucson, AZ 85721 USA. Univ Illinois, Community Architecture Network Informat Syst Lab, Urbana, IL 61820 USA. NCI, Int Canc Informat Ctr, Bethesda, MD 20852 USA. Univ Arizona, Sch Lib Sci, Tucson, AZ 85721 USA. Carnegie Mellon Univ, Sch Comp Sci, Pittsburgh, PA 15213 USA. RP Houston, AL (reprint author), Louisiana State Univ, Coll Business Adm, ISDS Dept, 3178B4 CEBA, Baton Rouge, LA 70803 USA. NR 21 TC 16 Z9 16 U1 0 U2 4 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0167-9236 J9 DECIS SUPPORT SYST JI Decis. Support Syst. PD DEC 27 PY 2000 VL 30 IS 2 BP 171 EP 186 DI 10.1016/S0167-9236(00)00097-X PG 16 WC Computer Science, Artificial Intelligence; Computer Science, Information Systems; Operations Research & Management Science SC Computer Science; Operations Research & Management Science GA 377ZN UT WOS:000165556500006 ER PT J AU Arinami, T Ishiguro, H Onaivi, ES AF Arinami, T Ishiguro, H Onaivi, ES TI Polymorphisms in genes involved in neurotransmission in relation to smoking SO EUROPEAN JOURNAL OF PHARMACOLOGY LA English DT Review DE smoking; genetic polymorphism; neurotransmitter; association; linkage ID CATECHOL-O-METHYLTRANSFERASE; SEROTONIN TRANSPORTER GENE; MONOAMINE-OXIDASE-A; TRYPTOPHAN-HYDROXYLASE GENE; DOPAMINE-RECEPTOR GENE; AMINO-ACID SUBSTITUTION; OBSESSIVE-COMPULSIVE DISORDER; CYTOCHROME-P450 2A6 CYP2A6; HUMAN PERSONALITY-TRAIT; LOW-ACTIVITY ALLELE AB Smoking behavior is influenced by both genetic and environmental factors. The genetic contribution to smoking behavior is at least as great as its contribution to alcoholism. Much progress has been achieved in genomic research related to cigarette-smoking within recent years. Linkage studies indicate that there are several loci linked to smoking, and candidate genes that are related to neurotransmission have been examined. Possible associated genes include cytochrome P450 subfamily polypeptide 6 (CYP2A6), dopamine D-1, D-2. and D-4 receptors, dopamine transporter, and serotonin transporter genes. There are other important candidate genes but studies evaluating the link with smoking have not been reported. These include genes encoding the dopamine D-3 and D-5 receptors, serotonin receptors, tyrosine hydroxylase, trytophan 2,3-dioxygenase, opioid receptors, and cannabinoid receptors. Since smoking-related factors are extremely complex, studies of diverse populations and of many aspects of smoking behavior including initiation, maintenance, cessation, relapse, and influence of environmental factors are needed to identify smoking-associated genes. We now review genetic polymorphisms reported to be involved in neurotransmission in relation to smoking. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Univ Tsukuba, Inst Basic Med Sci, Dept Med Genet, Tsukuba, Ibaraki 3058575, Japan. NIDA, Intramural Res Program, NIH, Baltimore, MD 21224 USA. RP Arinami, T (reprint author), Univ Tsukuba, Inst Basic Med Sci, Dept Med Genet, Tsukuba, Ibaraki 3058575, Japan. NR 158 TC 51 Z9 54 U1 6 U2 7 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0014-2999 J9 EUR J PHARMACOL JI Eur. J. Pharmacol. PD DEC 27 PY 2000 VL 410 IS 2-3 BP 215 EP 226 DI 10.1016/S0014-2999(00)00816-5 PG 12 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 391CW UT WOS:000166337400010 PM 11134671 ER PT J AU Snider, LA Swedo, SE AF Snider, LA Swedo, SE TI Pediatric obsessive-compulsive disorder SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Editorial Material ID CHILDHOOD; ADOLESCENTS; CHILDREN; SYMPTOMS; OCD C1 NIMH, Pediat & Dev Neuropsychiat Branch, NIH, Bethesda, MD 20892 USA. RP Snider, LA (reprint author), NIMH, Pediat & Dev Neuropsychiat Branch, NIH, 10 Ctr dr,MSC 1255,Bldg 10,Room 4N208, Bethesda, MD 20892 USA. NR 23 TC 17 Z9 17 U1 0 U2 3 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD DEC 27 PY 2000 VL 284 IS 24 BP 3104 EP 3106 DI 10.1001/jama.284.24.3104 PG 3 WC Medicine, General & Internal SC General & Internal Medicine GA 385FZ UT WOS:000165994100002 PM 11135753 ER PT J AU Troisi, RJ Cowie, CC Harris, MI AF Troisi, RJ Cowie, CC Harris, MI TI Diurnal variation in pasting plasma glucose - Implications for diagnosis of diabetes in patients examined in the afternoon SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID GROWTH-HORMONE SECRETION; DAWN PHENOMENON; INSULIN; MELLITUS; PATTERNS; ROLES; SLEEP AB Context Current diagnostic criteria for diabetes are based on plasma glucose levels in blood samples obtained in the morning after an overnight fast, with a value of 7.0 mmol/L (126 mg/dL) or more indicating diabetes. However, many patients are seen by their physicians in the afternoon. Because plasma glucose levels are higher in the morning, it is unclear whether these diagnostic criteria can be applied to patients who are tested for diabetes in the afternoon. Objectives To document diurnal variation in fasting plasma glucose levels in adults not known to have diabetes, and to examine the applicability to afternoon-examined patients of the current diagnostic criteria for diabetes. Design, Setting, and Participants Analysis of data from the US population-based Third National Health and Nutrition Examination Survey (1988-1994) on participants aged 20 years or older who had no previously diagnosed diabetes, who were randomly assigned to morning (n=6483) or afternoon (n=6399) examinations, and who fasted prior to blood sampling. Main Outcome Measures Fasting plasma glucose levels in morning- vs afternoon-examined participants; diabetes diagnostic value for afternoon-examined participants. Results The morning and afternoon groups did not differ in age, body mass index, waist-to-hip ratio, physical activity index, glycosylated hemoglobin level, and other factors. Mean (SD) fasting plasma glucose levels were higher in the morning group (5.41 [0.01] mmol/L [97.4 {0.3} mg/dL]) than in the afternoon group (5.12 [0.02] mmol/L [92.4 {0.4} mg/dL]; P<.001). Consequently, prevalence of afternoon-examined participants with fasting plasma glucose levels of 7.0 mmol/L (126 mg/dL) or greater was half that of participants examined in the morning. The diagnostic fasting plasma glucose value for afternoon-examined participants that resulted in the same prevalence of diabetes found in moming-examined participants was 6.33 mmol/L (114 mg/dL) or greater. Conclusions Our results indicate that if current diabetes diagnostic criteria are applied to patients seen in the afternoon, approximately half of all cases of undiagnosed diabetes in these patients will be missed. C1 NIDDKD, Bethesda, MD 20892 USA. Social & Sci Syst Inc, Bethesda, MD USA. RP Harris, MI (reprint author), NIDDKD, Room 695,6707 Democracy Blvd,MSC-5460, Bethesda, MD 20892 USA. NR 13 TC 63 Z9 64 U1 0 U2 0 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD DEC 27 PY 2000 VL 284 IS 24 BP 3157 EP 3159 DI 10.1001/jama.284.24.3157 PG 3 WC Medicine, General & Internal SC General & Internal Medicine GA 385FZ UT WOS:000165994100034 PM 11135780 ER PT J AU Lakshman, MK Kole, PL Chaturvedi, S Saugier, JH Yeh, HJC Glusker, JP Carrell, HL Katz, AK Afshar, CE Dashwood, WM Kenniston, G Baird, WM AF Lakshman, MK Kole, PL Chaturvedi, S Saugier, JH Yeh, HJC Glusker, JP Carrell, HL Katz, AK Afshar, CE Dashwood, WM Kenniston, G Baird, WM TI Methyl group-induced helicity in 1,4-dimethylbenzo[c]phenanthrene and its metabolites: Synthesis, physical, and biological properties SO JOURNAL OF THE AMERICAN CHEMICAL SOCIETY LA English DT Article ID REGION DIOL EPOXIDES; DNA-ADDUCTS; 11,12-DIHYDRODIOL 13,14-EPOXIDE; BENZOPHENANTHRENE; PHOTOCYCLIZATION; 1,2-EPOXIDES; DERIVATIVES; ADENINE AB 1,4-Dimethylbenzo[c]phenanthrene (1,4-DMBcPh) is the dimethylated analogue of the benzo[c]phenanthrene (BcPh), where one of its two methyl groups resides in the highly congested fjord-region. A comparative X-ray crystallographic analysis, described herein, shows that BcPh is distorted out-of-plane so that there is an angle of 27 degrees between the outermost rings. The additional methyl groups in I,4-DMBcPh increase this nonplanarity to an angle of 37 degrees. This methyl group-induced disruption of planarity results in P and M enantiomers of 1,4-DMBcPh, and this helicity is observed in a pronounced manner in its putative metabolites, the dihydrodiol and diol epoxide. Synthetically, photochemical cyclization offers convenient access to 1,4-DMBcPh as well as its metabolites. For this, Wittig reaction of 2,5-dimethylbenzyltriphenylphosphoninm chloride and 2-naphthaldehyde provided a cis/trans mixture of alkenes which, when subjected to photolysis, provided],4-DMBcPh. Despite the high steric congestion in the fjord-region, this reaction proceeds with respectable yields. Correspondingly, a Wittig reaction of the same phosphonium chloride derivative with 6-methoxy-2-naphthaldehyde afforded an alkene mixture that, upon photochemical ring closure, provided 10-methoxy-1,4-dimethylbenzo[c]phenanthrene. The methoxy group was cleaved and the resulting phenol oxidized to the o-quinone. Reduction of the o-quinone with NaBH4 under an oxygen atmosphere provided the dihydrodiol, (+/-)-trans-9, 10-dihydroxy-9, 10-dihydro-1,4-dimethylbenzo[c]phenanthrene. This dihydrodiol was found to be an approximately 3:1 mixture of diastereomers, which was produced as follows. Reduction of the quinone proceeds in a stereoselective manner producing both the (R,R)- and (S,S)-trans-diols. However, this factor, when coupled with the P, M atropisomerism of the hydrocarbon, results in two diastereomeric pairs of enantiomeric dihydrodiols. Due to steric constraints imposed by the fjord-region methyl group, the P --> M interconversion is slow, making the proton resonances of the diastereomeric dihydrodiols distinctly observable by NMR. Assignment of major and minor dihydrodiol isomers has been achieved by NOESY experiments. Finally, epoxidation provides a mixture of diol epoxides that reflects the dihydrodiol ratio. The metabolic activation of these compounds to reactive intermediates was studied through analysis of their binding to DNA. DNA binding data using human mammary carcinoma MCF-7 cells reveal that the level of DNA binding of BcPh is not statistically different from that of 1,4-DMBcPh. However, there is an Ii-fold increased activation of BcPh dihydrodiol as compared to the 1,4-DMBcPh dihydrodiol. In contrast to the planar benzo[a]pyrene, BcPh is only poorly adducted to DNA in culture cells. Thus, it appears that increasing the nonplanarity in this type of PAH lowers its ability to be metabolically activated to form DNA-damaging adducts, although in the case of 1,4-DMBcPh, the presence of the two methyl groups in one of the angular rings may also contribute to the decrease. C1 Univ N Dakota, Dept Chem, Grand Forks, ND 58202 USA. Chemsyn Sci Labs, Lenexa, KS 66215 USA. PE Biosyst, Foster City, CA 94404 USA. NIDDK, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. Fox Chase Canc Ctr, Inst Canc Res, Philadelphia, PA 19111 USA. Oregon State Univ, Environm Hlth Sci Ctr, Corvallis, OR 97331 USA. RP Lakshman, MK (reprint author), CUNY City Coll, Dept Chem, 138th St & Convent Ave, New York, NY 10031 USA. NR 46 TC 35 Z9 36 U1 0 U2 2 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0002-7863 J9 J AM CHEM SOC JI J. Am. Chem. Soc. PD DEC 27 PY 2000 VL 122 IS 51 BP 12629 EP 12636 DI 10.1021/ja002072w PG 8 WC Chemistry, Multidisciplinary SC Chemistry GA 386DN UT WOS:000166045000003 ER PT J AU Senderowicz, AM AF Senderowicz, AM TI Small molecule modulators of cyclin-dependent kinases for cancer therapy SO ONCOGENE LA English DT Article DE cell cycle; flavopiridol; UCN-01; cyclin-dependent kinases; cyclin D1; protein kinases; apoptosis ID BREAST-CARCINOMA CELLS; FLAVOPIRIDOL INDUCES APOPTOSIS; CHRONIC LYMPHOCYTIC-LEUKEMIA; HUMAN EPIDERMOID CARCINOMA; PROTEIN-KINASE; SELECTIVE INHIBITOR; 7-HYDROXYSTAUROSPORINE UCN-01; ANTITUMOR-ACTIVITY; IN-VITRO; RADIOSENSITIZING AGENT AB The majority of human malignancies have aberrancies in the Retinoblastoma (Rb) pathway, Loss in Rb function results from the phosphorylation and inactivation of Rb by the cyclin-dependent kinases (cdks), main regulators of cell cycle progression, Thus, modulators of cdks mag have a role in the treatment of human malignancies. Flavopiridol, the first cdk modulator tested in clinical trials, demonstrates interesting preclinical features: cell cycle block, induction of apoptosis, promotion of differentiation, inhibition of angiogenic processes and modulation of transcriptional events. Initial clinical trials with infusional flavopiridol demonstrated activity in some patients with lymphomas and renal, colon gastric carcinomas. Main side effects were diarrhea and hypotension. Phase 2 trials with infusional flavopiridol, other schedules and combination with standard chemotherapies are ongoing, The second cdk modulator tested in clinical trials, UCN-01, is a PKC inhibitor that can also modulate cdk activity. Similar to flavopiridol, UCN-01 blocks cell cycle progression and promotes apoptosis. Moreover, UCN-01 may abrogate checkpoints induced by genotoxic stress due to inhibition of chk1 kinase. The first clinical trial of UCN-01 demonstrated very prolonged half-life (similar to 600 h), due to high binding affinity of UCN-01 to the human alpha-1-acid glycoprotein. Main side effects mere headaches, vomiting, hypoxemia and hyperglycemia. Clinical activity was observed in some patients with melanoma and lymphoma. Trials of shorter infusions of UCN-01 or in combination with standard chemotherapeutic agents are ongoing. Although several important basic and clinical questions remain unanswered, development of cdk modulators is a reasonable strategy for cancer therapy. C1 NIDCR, Mol Therapeut Unit, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD 20892 USA. RP Senderowicz, AM (reprint author), NIDCR, Mol Therapeut Unit, Oral & Pharyngeal Canc Branch, NIH, Bldg 30,Room 211,30 Convent Dr, Bethesda, MD 20892 USA. NR 77 TC 96 Z9 97 U1 1 U2 11 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD DEC 27 PY 2000 VL 19 IS 56 BP 6600 EP 6606 DI 10.1038/sj.onc.1204085 PG 7 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 415CY UT WOS:000167704000006 PM 11426645 ER PT J AU Kuznetsov, SA Mankani, MH Robey, PG AF Kuznetsov, SA Mankani, MH Robey, PG TI Effect of serum on human bone marrow stromal cells: Ex vivo expansion and in vivo bone formation SO TRANSPLANTATION LA English DT Article ID MESENCHYMAL STEM-CELLS; MICRO-ENVIRONMENT TRANSFER; SKELETAL PROGENITOR CELLS; FORMATION IN-VITRO; OSTEOGENIC PRECURSORS; FIBROBLASTS; TRANSPLANTATION; REQUIREMENTS; CULTURES; CAPACITY AB Background. Bone marrow stromal cell (BMSC) transplantation may offer an efficacious method for the repair of bone defects. This approach has been developed using BMSCs expanded ex vivo in medium with fetal bovine serum (FBS), For clinical applications, however, contact of BMSCs with FBS should be minimized. We studied the effect of FBS substitutes on both human BMSC proliferation in vitro and subsequent bone formation in vivo. Methods. BMSC proliferation was measured by colony forming efficiency (CFE) and by cell numbers at consecutive passages. Bone formation was studied in 6- to 8-week-old transplants of human BMSCs in immunocompromised mice. Results. Medium with FBS was more effective in stimulating BMSC proliferation than medium with either human serum (HS) or rabbit serum (RS), Compared to bone formed by BMSCs cultured continuously with FBS, bone formed by cells cultured with HS, or with FBS switched to HS, was considerably less extensive, while bone formed by cells cultured with FBS witched to serum-free medium (SFM) was considerably more extensive. The increase in bone formation was due to neither the SFM components nor to the proliferation status of BMSCs prior to transplantation. Conclusions. Our data demonstrate that for ex vivo expansion of human BMSCs, medium with FBS remains most effective. However, incubation of human BMSCs in SFM prior to in vivo transplantation significantly stimulates subsequent bone formation. This finding increases the practicality of using culture-expanded BMSCs for autologous human transplantation and suggests the presence of osteogenic inhibitors in serum. C1 Natl Inst Dent & Craniofacial Res, Craniofacial & Skeletal Dis Branch, NIH, Bethesda, MD 20892 USA. RP Kuznetsov, SA (reprint author), Natl Inst Dent & Craniofacial Res, Craniofacial & Skeletal Dis Branch, NIH, Bldg 30,Room 228,30 Convent Dr MSC 4320, Bethesda, MD 20892 USA. RI Robey, Pamela/H-1429-2011 OI Robey, Pamela/0000-0002-5316-5576 NR 39 TC 111 Z9 122 U1 1 U2 6 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0041-1337 J9 TRANSPLANTATION JI Transplantation PD DEC 27 PY 2000 VL 70 IS 12 BP 1780 EP 1787 DI 10.1097/00007890-200012270-00018 PG 8 WC Immunology; Surgery; Transplantation SC Immunology; Surgery; Transplantation GA 388WN UT WOS:000166206200018 PM 11152111 ER PT J AU Sergeev, YV Wingfield, PT Hejtmancik, JF AF Sergeev, YV Wingfield, PT Hejtmancik, JF TI Monomer-dimer equilibrium of normal and modified beta A3-crystallins: Experimental determination and molecular modeling SO BIOCHEMISTRY LA English DT Article ID C-TERMINAL EXTENSIONS; LENS BETA-CRYSTALLIN; X-RAY-ANALYSIS; EYE-LENS; GAMMA-CRYSTALLIN; PROTEIN ASSOCIATION; CATARACT; GENE; EVOLUTION; MUTATION AB beta- and gamma -Crystallins are major protein constituents of the mammalian lens, where their stability and association into higher order complexes are critical for clarity and refraction. Two regions of the beta gamma -crystallins have been suggested to modulate protein association, namely, the flexible N-terminal extensions and the intramolecular domain interfaces. The oligomeric state of wild-type recombinant murine beta A3-crystallin (r beta A3) was compared to that of modified beta A3-crystallins with either an N-terminal deletion of residues 1 to 29 (r beta A3tr) or with residues 114 to 123 of the interdomain linker replaced with the analogous linker from murine gammaB-crystallin (r beta A3cp). All three proteins exhibited reversible monomer-dimer formation. The modifications to the N-terminus and domain linker resulted in tighter dimer formation as compared to wild-type protein as indicated by disassociation constants determined by sedimentation equilibrium: 6.62 x 10(-6) M (r beta A3), 0.86 x 10(-6) M (r beta A3cp), and 1.83 x 10(-7) M (r beta A3tr). Homology modeling of beta A3-crystallins and solvation energy calculations also predicted tighter binding of the modified crystallins consistent with the centrifugation results. The findings suggest that under physiological conditions beta A3 crystallin exists in a dynamic equilibrium between monomeric and dimeric protein and that modification, especially to the N-terminal extension, can promote self-association. C1 NEI, OGVFB, NIH, Bethesda, MD 20892 USA. NIAMSD, NIH, Bethesda, MD 20892 USA. RP NEI, OGVFB, NIH, 10-10B10, Bethesda, MD 20892 USA. EM sergeev@helix.nih.gov NR 50 TC 27 Z9 29 U1 0 U2 4 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD DEC 26 PY 2000 VL 39 IS 51 BP 15799 EP 15806 DI 10.1021/bi001882h PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 386PD UT WOS:000166070000017 PM 11123905 ER PT J AU Pendrak, ML Krutzsch, HC Roberts, DD AF Pendrak, ML Krutzsch, HC Roberts, DD TI Structural requirements for hemoglobin to induce fibronectin receptor expression in Candida albicans SO BIOCHEMISTRY LA English DT Article ID LIGAND-BINDING SYSTEMS; HAEMOPHILUS-INFLUENZAE; IRON ACQUISITION; HEMOLYTIC FACTOR; HEME-BINDING; PROTEIN; VIRULENCE; PATHOGENESIS; HAPTOGLOBIN; INFECTIONS AB Hemoglobin (Hb) is a host factor that induces expression of a promiscuous receptor on Candida albicans for fibronectin (FN) and several other extracellular matrix proteins. FN receptor expression was induced by ferric (Hb(+)Met and Hb(+)CN), ferrous (HbCO and HbO(2)), and cobalt-protoporphyrin derivatives of Hb, whereas globin was inactive. The Hb derivatives all exhibited saturable, dose-dependent kinetics of FN receptor induction, suggesting that Hb may be acting as a receptor ligand. Soluble Hb bound saturably to a low-affinity binding site [K-d = (1.1 +/- 0.2) X 10(-6) M] on C. albicans blastospores. However, uptake of (55)FeHb revealed that heme or iron transport into the cell is not required for induction, since internalization of Fe-55 from Hb did not occur until after induction of FN binding. The serum Hb-binding protein, haptoglobin, specifically abrogated this response, indicating that protein structure rather than the heme ligand or iron is necessary for induction of this signaling pathway. C. albicans also adhered to immobilized Hb, which was sufficient to induce FN receptor expression, and to Hb polymers that formed in defined Hb liquid media in the presence of cells. Formation of Hb polymers in solution required metabolic energy, since the aggregation process was halted with azide addition. Collectively, these data demonstrate that C. albicans recognizes polymerized Hb through multivalent low-affinity interactions, and this may be a host environmental cue that triggers extracellular matrix receptor expression at a septic site. C1 NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. RP Roberts, DD (reprint author), NCI, Pathol Lab, NIH, Bldg 10,Room 2A33,10 Ctr Dr,MSC 1500, Bethesda, MD 20892 USA. RI Roberts, David/A-9699-2008 OI Roberts, David/0000-0002-2481-2981 NR 57 TC 21 Z9 21 U1 0 U2 0 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD DEC 26 PY 2000 VL 39 IS 51 BP 16110 EP 16118 DI 10.1021/bi0012585 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 386PD UT WOS:000166070000051 PM 11123939 ER PT J AU Yang, SA Klee, CB AF Yang, SA Klee, CB TI Low affinity Ca2+-binding sites of calcineurin B mediate conformational changes in calcineurin A SO BIOCHEMISTRY LA English DT Article ID STIMULATED PROTEIN PHOSPHATASE; AUTOINHIBITORY DOMAIN; LIMITED PROTEOLYSIS; CALMODULIN; COMPLEX; ACTIVATION; SUBUNIT; LIBRARIES; TARGET; PHAGE AB Limited proteolysis of calcineurin in the presence of Ca2+ suggested that its calmodulin-binding domain, readily degraded by proteases, was unfolded while calcineurin B was compactly folded [Hubbard, M. J., and Klee, C. B. (1989) Biochemistry 28, 1868-1874]. Moreover, in the crystal structure of calcineurin, with the four Ca2+ sites of calcineurin B occupied, the calmodulin-binding domain is not visible in the electron density map [Kissinger, C. R., et al. (1995) Nature 378, 641-644]. Limited proteolysis of calcineurin in the presence of EGTA, shows that, when the low affinity sites of calcineurin B are not occupied, the calmodulin-binding domain is completely protected against proteolytic attack. Slow cleavages are, however, detected in the linker region between the calmodulin-binding and the autoinhibitory domains of calcineurin A. Upon prolonged exposure to the protease, selective cleavages in carboxyl-terminal end of the first helix and the central helix linker of calcineurin B and the calcineurin B-binding helix of calcineurin A are also detected. Thus, Ca2+ binding to the low-affinity sites of calcineurin B affects the conformation of calcineurin B and induces a conformational change of the regulatory domain of calcineurin A, resulting in the exposure of the calmodulin-binding domain. This conformational change is needed for the partial activation of the enzyme in the absence of calmodulin and its full activation by calmodulin. A synthetic peptide corresponding to the calmodulin-binding domain is shown to interact with a peptide corresponding to the calcineurin B-binding domain, and this interaction is prevented by calcineurin B in the presence but not the absence of Ca2+. These observations provide a mechanism to explain the dependence on Ca2+ binding to calcineurin B for calmodulin activation and for the 10-20-fold increase in affinity of calcineurin for Ca2+ upon removal of the regulatory domain by limited proteolysis [Stemmer, P. M., and Klee, C. B. (1994) Biochemistry 33, 6859-6866]. C1 NCI, Biochem Lab, NIH, Bethesda, MD 20892 USA. RP Klee, CB (reprint author), NCI, Biochem Lab, NIH, 37 Convent Dr,Bldg 37,Room 4E28, Bethesda, MD 20892 USA. NR 36 TC 55 Z9 56 U1 0 U2 5 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD DEC 26 PY 2000 VL 39 IS 51 BP 16147 EP 16154 DI 10.1021/bi001321q PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 386PD UT WOS:000166070000055 PM 11123943 ER PT J AU Bara-Jimenez, W Shelton, P Hallett, M AF Bara-Jimenez, W Shelton, P Hallett, M TI Spatial discrimination is abnormal in focal hand dystonia SO NEUROLOGY LA English DT Article ID PRIMARY SOMATOSENSORY CORTEX; REPETITIVE STRAIN INJURY; TEMPORAL DISCRIMINATION; WRITERS CRAMP; REPRESENTATION; RESOLUTION; INHIBITION; HUMANS; MODEL AB Background: In patients with focal hand dystonia, abnormal digit representations in the primary somatosensory cortex (S1) could be the result of enlarged and overlapping receptor fields, as suggested by an animal model of dystonia. A possible clinical correlate of this S1 abnormality is a disturbed spatial discrimination capability. Objective: To test the hypothesis that somatosensory spatial discrimination is abnormal in focal hand dystonia. Methods: Seventeen patients with focal hand dystonia underwent a quantitative evaluation of somatosensory spatial frequency (gap detection, JVP domes, applied to the distal phalanx of the index finger) and single-touch localization (Von Frey monofilaments, applied to the middle phalanx of the index finger). Results: Compared with control subjects, patients had a decreased performance in both the gap detection (p = 0.004) and the localization (p = 0.013) tasks. The extent of spatial frequency abnormality correlated with age in both groups. Conclusions: These findings, together with a previously shown temporal discrimination deficit, support a role for sensory dysfunction in the pathophysiology of dystonia. C1 NINDS, Human Motor Control Sect, Med Neurol Branch, NIH, Bethesda, MD 20892 USA. RP Hallett, M (reprint author), NINDS, Human Motor Control Sect, Med Neurol Branch, NIH, Bldg 10,Rm 5N226,10 Ctr Dr,MSC 1428, Bethesda, MD 20892 USA. NR 26 TC 110 Z9 113 U1 3 U2 5 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0028-3878 J9 NEUROLOGY JI Neurology PD DEC 26 PY 2000 VL 55 IS 12 BP 1869 EP 1873 PG 5 WC Clinical Neurology SC Neurosciences & Neurology GA 385TB UT WOS:000166018800020 PM 11134387 ER PT J AU Tezak, Z Nagaraju, K Plotz, P Hoffman, EP AF Tezak, Z Nagaraju, K Plotz, P Hoffman, EP TI Adeno-associated virus in normal and myositis human skeletal muscle SO NEUROLOGY LA English DT Article ID ADENOASSOCIATED VIRUS; TYPE-2 AB The normal tissue tropism of adeno-associated virus (AAV) is poorly defined, although the majority of humans test seropositive for this virus. Eighty-five muscle biopsy specimens were tested for AAV genomes; AAV DNA was identified in 17% of normal and 10% of Duchenne muscular dystrophy muscle biopsy specimens, but in only 3% of peripheral blood samples. AAV genomes were absent from all 37 muscle biopsy specimens from patients with myositis tested. Muscle is a major target organ for AAV, and infection is associated with autoimmune disease of muscle. C1 George Washington Univ, Childrens Natl Med Ctr, Med Genet Res Ctr, Washington, DC 20010 USA. NIAMSD, Arthrit & Rheumatism Branch, NIH, Bethesda, MD 20892 USA. Univ Pittsburgh, Sch Med, Dept Mol Genet & Biochem, Pittsburgh, PA 15261 USA. RP Hoffman, EP (reprint author), George Washington Univ, Childrens Natl Med Ctr, Med Genet Res Ctr, 111 Michigan Ave NW, Washington, DC 20010 USA. FU NINDS NIH HHS [NS29525-09] NR 10 TC 6 Z9 7 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0028-3878 J9 NEUROLOGY JI Neurology PD DEC 26 PY 2000 VL 55 IS 12 BP 1913 EP 1917 PG 5 WC Clinical Neurology SC Neurosciences & Neurology GA 385TB UT WOS:000166018800030 PM 11134397 ER PT J AU Munoz, A Petering, DH Shaw, CF AF Munoz, A Petering, DH Shaw, CF TI Structure-reactivity relationships among metallothionein three-metal domains: Role of non-cysteine amino acid residues in lobster metallothionein and human metallothionein-3 SO INORGANIC CHEMISTRY LA English DT Article ID MAGNETIC-RESONANCE SPECTROSCOPY; RABBIT LIVER METALLOTHIONEIN; DISULFIDE INTERCHANGE REACTIONS; GROWTH-INHIBITORY FACTOR; 3-DIMENSIONAL STRUCTURE; BETA-DOMAINS; ALPHA-DOMAIN; CLUSTERS; KINETICS; DTNB AB Metallothionlin (MT) domains of different origins,exhibiting distinct, highly conserved cysteine positions, show differences in metal-cysteine coordination and reactivity. Lobster MT, which includes two Cd3S9 beta domains, was chosen as a basic model to study the structure-function relationship among the clusters. The possible influence of the position of the cysteine residues and (2) the steric and electrostatic effects of neighboring amino acids on the folding and stability of MT clusters have been examined with the native lobster beta (c) and beta (N) domains, each having nine cysteines and binding three M2+ ions, and a modified domain beta (C-N), in which the cysteines of the C-terminal domain are relocated so they are spaced as in the N-terminal domain. Each has been synthesized and characterized by UV, CD,Cd-113 NMR, and H-1 NMR spectroscopies. The synthetic native domains (Cd(3)beta (c) and Cd(3)beta (N)) displayed spectroscopic properties, metal-binding affinities, and kinetic reactivity similar to those of the hole protein. In contrast, the modified Cd(3)beta (C-N) domain was unusually reactive and, in the presence of Chelax, a metal-ion chelating resin, was converted to a Cd-5(beta (C-N))(2) dimer. These differences in structure and reactivity demonstrate that the requirements for formation of a stable type-B, Cd3S9, beta cluster are more stringent than simply the sequential positions of the cysteines along the peptide chain and include specific interactions with neighboring amino acids. Molecular mechanics calculations suggest that changes of even a single amino acid in lobster Cd(3)beta (N) toward lobster Cd(3)beta (C-->N) dr in mammalian MT1 or MT2 toward Cd(3)beta -MT3 (GIF) can destabilize their structures. C1 Univ Wisconsin, Dept Chem, Milwaukee, WI 53201 USA. Univ Wisconsin, UWM, NIEHS Marine, Milwaukee, WI 53201 USA. Univ Wisconsin, Freshwater Biomed Sci Ctr, Milwaukee, WI 53201 USA. Eastern Kentucky Univ, Dept Chem, Richmond, KY 40475 USA. RP Petering, DH (reprint author), Univ Wisconsin, Dept Chem, POB 413, Milwaukee, WI 53201 USA. FU NIDDK NIH HHS [DK 51308]; NIEHS NIH HHS [ES 04026, ES 04184] NR 55 TC 9 Z9 11 U1 0 U2 4 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0020-1669 J9 INORG CHEM JI Inorg. Chem. PD DEC 25 PY 2000 VL 39 IS 26 BP 6114 EP 6123 DI 10.1021/ic000485s PG 10 WC Chemistry, Inorganic & Nuclear SC Chemistry GA 385TF UT WOS:000166019200032 PM 11188527 ER PT J AU Smith, JJ Evans, EK Murakami, M Moyer, MB Moseley, MA Woude, GV Kornbluth, S AF Smith, JJ Evans, EK Murakami, M Moyer, MB Moseley, MA Woude, GV Kornbluth, S TI Wee1-regulated apoptosis mediated by the Crk adaptor protein in Xenopus egg extracts SO JOURNAL OF CELL BIOLOGY LA English DT Article DE apoptosis; caspase; Crk; Wee1; Xenopus ID CELL-FREE APOPTOSIS; CYTOCHROME-C; DEPENDENT ACTIVATION; GRANULOSA-CELLS; WEE1 KINASE; GRANZYME-B; V-CRK; REQUIREMENT; BCL-2; MITOCHONDRIA AB Many of the biochemical reactions of apoptotic cell death, including mitochondrial cytochrome c release and caspase activation, can be reconstituted in cell-free extracts derived from Xenopus eggs. In addition, because caspase activation does not occur until the egg extract has been incubated for several hours on the bench, upstream signaling processes occurring before full apoptosis are rendered accessible to biochemical manipulation. We reported previously that the adaptor protein Crk is required for apoptotic signaling in egg extracts (Evans, E.K., W. Lu, S.L. Strum, B.J. Mayer, and S. Kornbluth. 1997. EMBO (Eur. Mel. Biol. Organ.) J. 16:230-241). Moreover, we demonstrated that removal of Crk Src homology (SH)2 or SH3 interactors from the extracts prevented apoptosis. We now report the finding that the relevant Crk SH2-interacting protein, important for apoptotic signaling in the extract, is the well-known cell cycle regulator, Wee1. We have demonstrated a specific interaction between tyrosine-phosphorylated Wee1 and the Crk SH2 domain and have shown that recombinant Wee1 can restore apoptosis to an extract depleted of SH2 interactors. Moreover, exogenous Wee1 accelerated apoptosis in egg extracts, and this acceleration was largely dependent on the presence of endogenous Crk protein. As other Cdk inhibitors, such as roscovitine and Myt1, did not act like Wee1 to accelerate apoptosis, we propose that Wee1-Crk complexes signal in a novel apoptotic pathway, which may be unrelated to Wee1's role as a cell cycle regulator. C1 Duke Univ, Med Ctr, Dept Pharmacol & Canc Biol, Durham, NC 27710 USA. NCI, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. Glaxo Wellcome Inc, Dept Struct Chem, Res Triangle Pk, NC 27709 USA. Van Andel Res Inst, Grand Rapids, MI 49503 USA. RP Kornbluth, S (reprint author), Duke Univ, Med Ctr, Dept Pharmacol & Canc Biol, C370 LSRC,Res Dr,Box 3813, Durham, NC 27710 USA. FU NIGMS NIH HHS [R01 GM56518, R01 GM056518] NR 59 TC 26 Z9 28 U1 1 U2 2 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD DEC 25 PY 2000 VL 151 IS 7 BP 1391 EP 1400 DI 10.1083/jcb.151.7.1391 PG 10 WC Cell Biology SC Cell Biology GA 389KM UT WOS:000166237200004 PM 11134069 ER PT J AU Danen, EHJ Sonneveld, P Sonnenberg, A Yamada, KM AF Danen, EHJ Sonneveld, P Sonnenberg, A Yamada, KM TI Dual stimulation of Ras/Mitogen-activated protein kinase and RhoA by cell adhesion to fibronectin supports growth factor-stimulated cell cycle progression SO JOURNAL OF CELL BIOLOGY LA English DT Article DE fibronectin; cell adhesion; G1 cell cycle; integrin; Rho ID ANCHORAGE-INDEPENDENT GROWTH; MAP KINASE; RAS TRANSFORMATION; BINDING PROTEIN; S-PHASE; FOCAL ADHESIONS; CDC42 GTPASES; GENE-PRODUCT; EXPRESSION; INTEGRIN AB In cellular transformation, activated forms of the small GTPases Ras and RhoA can cooperate to drive cells through the G1 phase of the cell cycle, Here, we show that a similar but substrate-regulated mechanism is involved in the anchorage-dependent proliferation of untransformed NIH-3T3 cells. Among several extracellular matrix components tested, only fibronectin supported growth factor-induced, E2F-dependent S phase entry. Although all substrates supported the mitogen-activated protein kinase (MAPK) response to growth factors, RhoA activity was specifically enhanced on fibronectin. Moreover, induction of cyclin D1 and suppression of p21(Cip/Waf) occurred specifically, in a Rho-dependent fashion, in cells attached to fibronectin. This ability of fibronectin to stimulate both Ras/MAPK- and RhoA-dependent signaling can explain its potent cooperation with growth factors in the stimulation of cell cycle progression. C1 Netherlands Canc Inst, Div Cell Biol, NL-1066 CX Amsterdam, Netherlands. Natl Inst Dent & Craniofacial Res, Craniofacial Dev Biol & Regenerat Branch, NIH, Bethesda, MD 20892 USA. RP Danen, EHJ (reprint author), Netherlands Canc Inst, Div Cell Biol, Plesmanlaan 121, NL-1066 CX Amsterdam, Netherlands. OI Yamada, Kenneth/0000-0003-1512-6805 NR 62 TC 89 Z9 91 U1 0 U2 0 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD DEC 25 PY 2000 VL 151 IS 7 BP 1413 EP 1422 DI 10.1083/jcb.151.7.1413 PG 10 WC Cell Biology SC Cell Biology GA 389KM UT WOS:000166237200006 PM 11134071 ER PT J AU Wang, H Parry, DAD Jones, LN Idler, WW Marekov, LN Steinert, PM AF Wang, H Parry, DAD Jones, LN Idler, WW Marekov, LN Steinert, PM TI In vitro assembly and structure of trichocyte keratin intermediate filaments: A novel role for stabilization by disulfide bonding SO JOURNAL OF CELL BIOLOGY LA English DT Article DE intermediate filaments; trichocyte keratins; cytokeratins; disulfide bonding; molecular organization ID HARD ALPHA-KERATIN; HAIR KERATINS; EPIDERMAL KERATIN; EXPRESSION; VIMENTIN; MOLECULES; PROTEINS; MOUSE; HETEROTETRAMERS; DIFFERENTIATION AB Intermediate filaments (IF) have been recognized as ubiquitous components of the cytoskeletons of eukaryotic cells for 25 yr. Historically the first IF proteins to be characterized were those from wool in the 1960s, when they were defined as low sulfur keratins derived from "microfibrils." These proteins are now known as the type Ia/type IIa trichocyte keratins that constitute keratin IF of several hardened epithelial cell types. However, to date, of the entire class of >40 IF proteins, the trichocyte keratins remain the only ones for which efficient in vitro assembly remains unavailable. In this paper, we describe the assembly of expressed mouse type Ia and type IIa trichocyte keratins into IF in high yield. In cross-linking experiments, we document that the alignments of molecules within reduced trichocyte IF are the same as in type Ib/IIb cytokeratins. However, when oxidized in vitro, several intermolecular disulfide bonds form and the molecular alignments rearrange into the pattern shown earlier by x-ray diffraction analyses of intact wool. We suggest the realignments occur because the disulfide bonds confer substantially increased stability to trichocyte keratin LE Our data suggest a novel role for disulfide bond cross linking in stabilization of these IF and the tissues containing them. C1 NIAMSD, Skin Biol Lab, NIH, Bethesda, MD 20892 USA. Massey Univ, Inst Fundamental Sci, Palmerston North 5301, New Zealand. CSIRO, Div Wool Technol, Geelong, Vic 3216, Australia. RP Steinert, PM (reprint author), NIAMS, NIH, Bldg 6 Room 425, Bethesda, MD 20892 USA. NR 44 TC 59 Z9 60 U1 1 U2 9 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD DEC 25 PY 2000 VL 151 IS 7 BP 1459 EP 1468 DI 10.1083/jcb.151.7.1459 PG 10 WC Cell Biology SC Cell Biology GA 389KM UT WOS:000166237200010 PM 11134075 ER PT J AU Golden, A Sadler, PL Wallenfang, MR Schumacher, JM Hamill, DR Bates, G Bowerman, B Seydoux, G Shakes, DC AF Golden, A Sadler, PL Wallenfang, MR Schumacher, JM Hamill, DR Bates, G Bowerman, B Seydoux, G Shakes, DC TI Metaphase to anaphase (mat) transition-defective Mutants in Caenorhabditis elegans SO JOURNAL OF CELL BIOLOGY LA English DT Article DE meiosis; metaphase arrest; cell cycle; C. elegans; anaphase promoting complex ID PROTEIN-PROTEIN INTERACTIONS; SENSITIVE-LETHAL MUTATIONS; SISTER-CHROMATID COHESION; C-ELEGANS; CHROMOSOME CONDENSATION; PROMOTING COMPLEX; HISTONE H3; GENETIC-ANALYSIS; CELL-DIVISION; MITOSIS AB The metaphase to anaphase transition is a critical stage of the eukaryotic cell cycle, and, thus, it is highly regulated. Errors during this transition can lead to chromosome segregation defects and death of the organism. In genetic screens for temperature-sensitive maternal effect embryonic lethal (Me1) mutants, we have identified 32 mutants in the nematode Caenorhabditis elegans in which fertilized embryos arrest as one-cell embryos. In these mutant embryos, the oocyte chromosomes arrest in metaphase of meiosis I without transitioning to anaphase or producing polar bodies. An additional block in M phase exit is evidenced by the failure to form pronuclei and the persistence of phosphohistone H3 and MPM-2 antibody staining. Spermatocyte meiosis is also perturbed; primary spermatocytes arrest in metaphase of meiosis I and fail to produce secondary spermatocytes. Analogous mitotic defects cause M phase delays in mitotic germline proliferation. We have named this class of mutants "mat" for metaphase to anaphase transition defective. These mutants, representing six different complementation groups, all map near genes that encode subunits of the anaphase promoting complex or cyclosome, and, here, we show that one of the genes, emb-27, encodes the C. elegans CDC16 ortholog. C1 Coll William & Mary, Dept Biol, Williamsburg, VA 23187 USA. NIDDKD, Lab Biochem & Genet, NIH, Bethesda, MD 20892 USA. Univ Houston, Dept Biol, Houston, TX 77204 USA. Johns Hopkins Univ, Sch Med, Dept Mol Biol & Genet, Baltimore, MD 21205 USA. Univ Texas, MD Anderson Canc Ctr, Dept Mol Genet, Houston, TX 77030 USA. Univ Oregon, Inst Mol Biol, Eugene, OR 97403 USA. RP Shakes, DC (reprint author), Coll William & Mary, Dept Biol, POB 8795, Williamsburg, VA 23187 USA. RI Schumacher, Jill/B-2932-2012 FU NIGMS NIH HHS [R15 GM060359, 1RO1GM58017-02, R01 GM058017] NR 70 TC 119 Z9 130 U1 0 U2 1 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD DEC 25 PY 2000 VL 151 IS 7 BP 1469 EP 1482 DI 10.1083/jcb.151.7.1469 PG 14 WC Cell Biology SC Cell Biology GA 389KM UT WOS:000166237200011 PM 11134076 ER PT J AU Makalowski, W AF Makalowski, W TI Genomic scrap yard: how genomes utilize all that junk SO GENE LA English DT Article; Proceedings Paper CT International Symposium on Evolution 2000 CY MAR 05-MAY 07, 2000 CL TOKYO, JAPAN DE exaptation; genome shuffling; genomic evolution; junk DNA; repetitive elements; retrogenes ID SELFISH DNA; GENE; EVOLUTION; SEQUENCE; INSERTION; ELEMENTS; RETROTRANSPOSITION; TRANSDUCTION; DROSOPHILA; PROMOTERS AB Interspersed repetitive sequences are major components of eukaryotic genomes. Repetitive elements comprise over 50% of the mammalian genome. Because the specific function of these elements remains to be defined and because of their unusual 'behavior' in the genome, they are often quoted as a selfish or junk DNA. Our view of the entire phenomenon of repetitive elements has to now be revised in light of data on their biology and evolution, especially in the light of what we know about the retroposons. I would like to argue that even if we cannot define the specific functions of these elements, we still can show that they are not useless pieces of the genomes. The repetitive elements interact with the whole genome and influence its evolution. Repetitive elements interact with the surrounding sequences and nearby genes. They may serve as recombination hot spots or acquire specific cellular functions such as RNA transcription control or even become part of protein coding regions. Finally, they provide very efficient mechanism for genomic shuffling. As such, repetitive elements should be called genomic scrap yard rather than junk DNA. Tables listings examples of recruited (exapted) transposable elements are available at http://www.ncbi.nlm.nih.gov/Makalowski/ScrapYard/. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIH, Natl Lib Med, Natl Ctr Biotechnol Informat, Bethesda, MD 20894 USA. RP Makalowski, W (reprint author), NIH, Natl Lib Med, Natl Ctr Biotechnol Informat, Bethesda, MD 20894 USA. EM makalowski@ncbi.nlm.nih.gov RI Makalowski, Wojciech/I-2843-2016 NR 31 TC 112 Z9 118 U1 2 U2 6 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-1119 J9 GENE JI Gene PD DEC 23 PY 2000 VL 259 IS 1-2 SI SI BP 61 EP 67 DI 10.1016/S0378-1119(00)00436-4 PG 7 WC Genetics & Heredity SC Genetics & Heredity GA 391DF UT WOS:000166338300009 PM 11163962 ER PT J AU Artlett, CM Ramos, R Jiminez, SA Patterson, K Miller, FW Rider, LG AF Artlett, CM Ramos, R Jiminez, SA Patterson, K Miller, FW Rider, LG CA Childhood Myositis Heterogeneity C TI Chimeric cells of maternal origin in juvenile idiopathic inflammatory myopathies SO LANCET LA English DT Article ID IDENTIFICATION; POLYMYOSITIS AB We identified maternal microchimerism by fluorescence in-situ hybridisation in magnetically-separated CD4 or CD8 peripheral blood cells of eight of nine male patients with juvenile idiopathic inflammatory myopathy, compared with two of nine healthy male controls. We also found maternal microchimerism in inflammatory lesions (one skin sample and nine muscle biopsy samples) of all ten patients examined, compared with two of ten biopsy samples from patients with other muscle disorders. These results suggest that maternal cells may be involved in the pathogenesis of juvenile idiopathic inflammatory myopathy. C1 Thomas Jefferson Univ, Div Rheumatol, Philadelphia, PA 19107 USA. Childrens Hosp & Reg Med Ctr, Dept Pathol, Seattle, WA USA. Univ Washington, Seattle, WA 98195 USA. US FDA, Ctr Biol Evaluat & Res, Div Monoclonal Antibodies, Bethesda, MD USA. NIAMSD, Arthrit & Rheumatism Branch, NIH, Bethesda, MD 20892 USA. RP Artlett, CM (reprint author), Thomas Jefferson Univ, Div Rheumatol, Philadelphia, PA 19107 USA. OI Rider, Lisa/0000-0002-6912-2458; Miller, Frederick/0000-0003-2831-9593 FU NIAMS NIH HHS [R01 AR19616, R29 AR45399] NR 5 TC 103 Z9 105 U1 0 U2 1 PU LANCET LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0140-6736 J9 LANCET JI Lancet PD DEC 23 PY 2000 VL 356 IS 9248 BP 2155 EP 2156 DI 10.1016/S0140-6736(00)03499-1 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 385HD UT WOS:000165997300016 PM 11191545 ER PT J AU Dumitru, CD Ceci, JD Tsatsanis, C Kontoyiannis, D Stamatakis, K Lin, JH Patriotis, C Jenkins, NA Copeland, NG Kollias, G Tsichlis, PN AF Dumitru, CD Ceci, JD Tsatsanis, C Kontoyiannis, D Stamatakis, K Lin, JH Patriotis, C Jenkins, NA Copeland, NG Kollias, G Tsichlis, PN TI TNF-alpha induction by LPS is regulated posttranscriptionally via a Tpl2/ERK-dependent pathway SO CELL LA English DT Article ID NF-KAPPA-B; ACTIVATED PROTEIN-KINASE; NECROSIS-FACTOR-ALPHA; T-CELL LINES; TRANSLATION INITIATION; SIGNALS ACTIVATION; ENDOTOXIC-SHOCK; INNATE IMMUNITY; MICE DEFICIENT; TPL-2 AB Tpl2 knockout mice produce tow levels of TNF-alpha when exposed to lipopolysaccharide (LPS) and they are resistant to LPS/D-Galactosamine-induced pathology. LPS stimulation of peritoneal macrophages from these mice did not activate MEK1, ERK1, and ERK2 but did activate JNK, p38 MAPK, and NF-kappaB. The block in ERK1 and ERK2 activation was causally linked to the defect in TNF-alpha induction by experiments showing that normal murine macrophages treated with the MEK inhibitor PD98059 exhibit a similar defect. Deletion of the AU-rich motif in the TNF-alpha mRNA minimized the effect of Tpl2 inactivation on the induction of TNF-alpha. Subcellular fractionation of LPS-stimulated macrophages revealed that LPS signals transduced by Tpl2 specifically promote the transport of TNF-alpha mRNA from the nucleus to the cytoplasm. C1 Thomas Jefferson Univ, Kimmel Canc Ctr, Dept Microbiol & Immunol, Philadelphia, PA 19107 USA. NCI, Mouse Canc Genet Program, FCRDC, Frederick, MD 21702 USA. Hellen Pasteur Inst, Genet Mol Lab, Athens 11521, Greece. RP Tsichlis, PN (reprint author), Thomas Jefferson Univ, Kimmel Canc Ctr, Dept Microbiol & Immunol, 233 S 10th St, Philadelphia, PA 19107 USA. RI Kollias, George/A-7079-2012; Stamatakis, Konstantinos/A-6596-2017 OI Kollias, George/0000-0003-1867-3150; Stamatakis, Konstantinos/0000-0001-9934-3502 FU NCI NIH HHS [R01 CA38047] NR 45 TC 522 Z9 539 U1 3 U2 22 PU CELL PRESS PI CAMBRIDGE PA 1050 MASSACHUSETTES AVE, CIRCULATION DEPT, CAMBRIDGE, MA 02138 USA SN 0092-8674 J9 CELL JI Cell PD DEC 22 PY 2000 VL 103 IS 7 BP 1071 EP 1083 DI 10.1016/S0092-8674(00)00210-5 PG 13 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 386BQ UT WOS:000166040600010 PM 11163183 ER PT J AU Wang, GS Peterkofsky, A Clore, GM AF Wang, GS Peterkofsky, A Clore, GM TI A novel membrane anchor function for the N-terminal amphipathic sequence of the signal-transducing protein IIA(Glucose) of the Escherichia coli phosphotransferase system SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID APOLIPOPROTEIN-A-I; SALMONELLA-TYPHIMURIUM; SECONDARY STRUCTURE; NMR-SPECTROSCOPY; SUGAR-TRANSPORT; IIIGLC; DOMAIN; IIA(GLC); PERMEASE; HELIX AB Enzyme IIA(Glucose) (IIA(Glc)) is a signal-transducing protein in the phosphotransferase system of Escherichia coli. Structural studies of free IIA(Glc) and the HPr-IIA(Glc) complex have shown that IIA(Glc) comprises a globular P-sheet sandwich core (residues 19-168) and a disordered N-terminal tail (residues 1-18). Although the presence of the N-terminal tail is not required for IIA(Glc) to accept a phosphorus from the histidine phosphocarrier protein HPr, its presence is essential for effective phosphotransfer from IIA(Glc) to the membrane-bound IIBCGlc. The sequence of the N-terminal tail suggests that it has the potential to form an amphipathic helix. Using CD, we demonstrate that a peptide, corresponding to the N-terminal 18 residues of IIA(Glc), adopts a helical conformation in the presence of either the anionic lipid phosphatidylglycerol or a mixture of anionic E. coli lipids phosphatidylglycerol (25%) and phosphatidylethanolamine (75%), The peptide, however, is in a random coil state in the presence of the zwitterionic lipid phosphatidylcholine, indicating that electrostatic interactions play a role in the binding of the lipid to the peptide. In addition, we show that intact IIA(Glc) also interacts with anionic lipids, resulting in an increase in helicity, which can be directly attributed to the N-terminal segment. From these data we propose that IIA(Glc) comprises two functional domains: a folded domain containing the active site and capable of weakly interacting with the peripheral IIB domain of the membrane protein IIBCGlc; and the N-terminal tail, which interacts with the negatively charged E, coli membrane, thereby stabilizing the complex of IIA(Glc) with IIBCGlc. This stabilization is essential for the final step of the phosphoryl transfer cascade in the glucose transport pathway. C1 NIDDK, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. NHLBI, Lab Biochem Genet, NIH, Bethesda, MD 20892 USA. RP Clore, GM (reprint author), NIDDK, Chem Phys Lab, NIH, Bldg 5,Rm B1-30I, Bethesda, MD 20892 USA. RI Clore, G. Marius/A-3511-2008 OI Clore, G. Marius/0000-0003-3809-1027 NR 26 TC 36 Z9 36 U1 0 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 22 PY 2000 VL 275 IS 51 BP 39811 EP 39814 DI 10.1074/jbc.C000709200 PG 4 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 386BD UT WOS:000166039500003 PM 11044440 ER PT J AU Bai, RL Covell, DG Pei, XF Ewell, JB Nguyen, NY Brossi, A Hamel, E AF Bai, RL Covell, DG Pei, XF Ewell, JB Nguyen, NY Brossi, A Hamel, E TI Mapping the binding site of colchicinoids on beta-tubulin - 2-chloroacetyl-2-demethylthiocolchicine covalently reacts predominantly with cysteine 239 and secondarily with cysteine 354 SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID BOVINE BRAIN; BIOLOGICAL PROPERTIES; NUCLEOCAPSID PROTEIN; INHIBITOR BINDING; THIOCOLCHICINE; AGENTS; IDENTIFICATION; SEPARATION; KINETICS; RESIDUE AB 2-Chloroacetyl-2-demethylthiocolchicine (2CTC) and 3-chloroacetyl-3-demethylthiocolchicine (3CTC) resemble colchicine in binding to tubulin and react covalently with beta -tubulin, forming adducts with cysteine residues 239 and 354. The adducts at Cys-239 are less stable than those at Cys-354 during formic acid digestion. Extrapolating to zero time, the Cys-239 to Cys-354 adduct ratio is 77:23 for 2CTC and 27:73 for 3CTC. Using energy minimization modeling to dock colchicinoids into the electron crystallographic model of P-tubulin in protofilaments (Nogales, E., Wolf, S. G., and Downing, K. H. (1998) Nature 391, 199-203), we found two potential binding sites. At one, entirely encompassed within beta -tubulin, the C2- and CS-oxygen atoms of 2CTC and 3CTC overlapped poorly with those of colchicine and thiocolchicine, but distances from the reactive carbon atoms of the analogs to the sulfur atoms of the cysteine residues were qualitatively consistent with reactivity. The other potential binding site was located at the alpha/beta interface. Here, the oxygen atoms of the analogs overlapped well with those of colchicine, but relative distances of the reactive carbons to the cysteine sulfur atoms did not correlate with the observed reactivity. A significant conformational change must occur in the colchicine binding site of tubulin in the transition from the unpolymerized to the polymerized state. C1 NCI, Screening Technol Branch, Dev Therapeut Program, Div Canc Treatment & Diag,Frederick Canc Res & De, Frederick, MD 21702 USA. NCI, Lab Expt & Computat Biol, Frederick Canc Res & Dev Ctr, Sci Applicat Int Corp, Frederick, MD 21702 USA. NIDDK, Struct Biol Lab, NIH, Bethesda, MD 20892 USA. US FDA, Ctr Biol Evaluat & Res, Facil Biotechnol Resources, Bethesda, MD 20892 USA. RP Hamel, E (reprint author), NCI, Screening Technol Branch, Dev Therapeut Program, Div Canc Treatment & Diag,Frederick Canc Res & De, Bldg 469,Rm 104,POB B, Frederick, MD 21702 USA. FU NCI NIH HHS [N01CO56000] NR 38 TC 72 Z9 78 U1 0 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 22 PY 2000 VL 275 IS 51 BP 40443 EP 40452 DI 10.1074/jbc.M005299200 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 386BD UT WOS:000166039500087 PM 11005811 ER PT J AU Gerelsaikhan, T Turner, RJ AF Gerelsaikhan, T Turner, RJ TI Transmembrane topology of the secretory Na+-K+-2Cl(-) cotransporter NKCC1 studied by in vitro translation SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID K-CL COTRANSPORTER; MEMBRANE-PROTEINS; MOLECULAR-CLONING; NA-K-2CL COTRANSPORTER; ENDOPLASMIC-RETICULUM; FUNCTIONAL EXPRESSION; HYPOKALEMIC ALKALOSIS; BUMETANIDE BINDING; BARTTERS-SYNDROME; PREDICTION AB The secretory Na+-K+-2Cl(-) cotransporter NKCC1 is a member of a small gene family of electroneutral salt transporters. Hydropathy analyses indicate that all of these transporters have a similar general structure consisting of large hydrophilic N and C termini on either side of a central, relatively well conserved, hydrophobic domain. Programs that predict the transmembrane topology of polytopic membrane proteins identify 10-12 putative membrane-spanning segments (MSSs) in this hydrophobic domain; but to date, there is little experimental data on the structure of this region for any of these transporters. In this report, we have studied the transmembrane topology of NKCC1 using an in vitro translation system designed to test the membrane insertion properties of putative MSSs (Bamberg, K., and Sachs, G. (1994) J. Biol. Chem. 269, 16909-16919). Fusion proteins consisting of putative NKCC1 MSSs inserted either (i) between an N-terminal cytosolic anchor sequence and a C-terminal reporter sequence containing multiple N-linked glycosidation sites or (ii) between an N-terminal signal anchor sequence and the same glycosidation flag were expressed in the presence of canine pancreatic microsomes. The glycosidation status of the reporter sequence, which indicated its luminal or extraluminal location in the microsomes, was then used to characterize the signal anchor or stop transfer activity of the inserted MSSs. The results of this experimental analysis yielded a topology scheme consisting of 12 membrane-spanning segments, two pairs of which apparently form rather tight hairpin-like structures within the membrane. C1 NIDCR, Membrane Biol Sect, Gene Therapy & Therapeut Branch, NIH, Bethesda, MD 20892 USA. RP Turner, RJ (reprint author), NIDCR, Membrane Biol Sect, Gene Therapy & Therapeut Branch, NIH, Rm 1A01,Bldg 10, Bethesda, MD 20892 USA. EM rjturner@nih.gov NR 37 TC 45 Z9 45 U1 1 U2 7 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 EI 1083-351X J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 22 PY 2000 VL 275 IS 51 BP 40471 EP 40477 DI 10.1074/jbc.M007751200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 386BD UT WOS:000166039500090 PM 11013260 ER PT J AU Sinal, CJ Miyata, M Tohkin, M Nagata, K Bend, JR Gonzalez, FJ AF Sinal, CJ Miyata, M Tohkin, M Nagata, K Bend, JR Gonzalez, FJ TI Targeted disruption of soluble epoxide hydrolase reveals a role in blood pressure regulation SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ARACHIDONIC-ACID EPOXYGENASE; NA-K-ATPASE; EPOXYEICOSATRIENOIC ACIDS; HYPERTENSIVE RATS; LIVER; METABOLITES; EICOSANOIDS; MICE; BIOSYNTHESIS; INHIBITORS AB Renal microsomal cytochrome P-450 monooxygenase-dependent metabolism of arachidonic acid generates a series of regioisomeric epoxyeicosatrienoic acids that can be further metabolized by soluble epoxide hydrolase to the corresponding dihydroxyeicosatrienoic acids. Evidence exists that these metabolites affect renal function and, in particular, blood pressure regulation. To examine this possibility, blood pressure and renal arachidonic acid metabolism were examined in mice with a targeted disruption of the soluble epoxide hydrolase gene. Systolic blood pressure of male soluble epoxide hydrolase-null mice was lower compared with wild-type mice in both the absence and presence of dietary salt loading. Both female soluble epoxide hydrolase-null and wild-type female mice also had significantly lower systolic blood pressure than male wild-type mice. Renal formation of epoxyeicosatrienoic and dihydroxyeicosatrienoic acids was markedly lower for soluble epoxide hydrolase-null versus wild-type mice of both sexes. Although disruption of soluble epoxide hydrolase in female mice had minimal effects on blood pressure, deletion of this gene feminized male mice by lowering systolic blood pressure and altering arachidonic acid metabolism. These data provide the first direct evidence for a role for soluble epoxide hydrolase in blood pressure regulation and identify this enzyme as a novel and attractive target for therapeutic intervention in hypertension. C1 NCI, Lab Metab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. Univ Western Ontario, Dept Pharmacol & Toxicol, London, ON N6A 5C1, Canada. RP Gonzalez, FJ (reprint author), NCI, Lab Metab, Div Basic Sci, NIH, Bldg 37,Rm 3E24, Bethesda, MD 20892 USA. NR 42 TC 229 Z9 235 U1 1 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 22 PY 2000 VL 275 IS 51 BP 40504 EP 40510 DI 10.1074/jbc.M008106200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 386BD UT WOS:000166039500095 PM 11001943 ER PT J AU Zheng, M Zhang, SJ Zhu, WZ Ziman, B Kobilka, BK Xiao, RP AF Zheng, M Zhang, SJ Zhu, WZ Ziman, B Kobilka, BK Xiao, RP TI beta(2)-adrenergic receptor-induced p38 MAPK activation is mediated by protein kinase A rather than by G(i) or G beta gamma in adult mouse cardiomyocytes SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID N-TERMINAL KINASES; CARDIAC MYOCYTES; TYROSINE KINASE; STRESS; HEART; HYPERTROPHY; STIMULATION; SPECIFICITY; INVOLVEMENT; PHOSPHATASE AB Increasing evidence shows that stimulation of p-adrenergic receptor (AR) activates mitogen-activated protein kinases (MAPKs), in addition to the classical G(s) adenylyl cyclase-cAMP-dependent protein kinase (PKA) signaling cascade. In the present study, we demonstrate a novel beta (2)-AR-mediated cross-talk between PKA and p38 MAPK in adult mouse cardiac myocytes expressing beta (2)-AR, with a null background of beta (1)beta (2)-AR double knockout. beta (2)-AR stimulation by isoproterenol increased p38 MARK activity in a time- and dose-dependent manner. Inhibiting G(i) with pertussis toxin or scavenging G beta gamma with beta ARK-ct overexpression could not prevent beta (2)-AR-induced p38 MAPK activation. In contrast, a specific peptide inhibitor of PKA, PKI (5 muM), completely abolished the stimulatory effect of beta (2)-AR, suggesting that beta (2)-AR-induced p38 MAPK activation is mediated via a PKA-dependent mechanism, rather than by G(i) or G beta gamma. This conclusion was further supported by the ability of forskolin (10 muM), an adenylyl cyclase activator, to elevate p38 MAPK activity in a PKI-sensitive manner. Furthermore, inhibition of p38 MAPK with SB203580 (10 muM) markedly enhanced the beta (2)-AR-mediated contractile response, without altering base-line contractility. These results provide the first evidence that cardiac beta (2)-AR activates p38 MAPK via a PKA-dependent signaling pathway, rather than by G(i) or G beta gamma, and reveal a novel role of p38 MAPK in regulating cardiac contractility. C1 NIA, Gerontol Res Ctr, Cardiovasc Sci Lab, NIH, Baltimore, MD 21224 USA. Stanford Univ, Med Ctr, Howard Hughes Med Inst, Stanford, CA 94305 USA. Peking Univ, Hosp 3, Sch Med, Inst Vasc Med, Beijing 100083, Peoples R China. RP Xiao, RP (reprint author), NIA, Gerontol Res Ctr, Cardiovasc Sci Lab, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 35 TC 95 Z9 98 U1 0 U2 8 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 22 PY 2000 VL 275 IS 51 BP 40635 EP 40640 DI 10.1074/jbc.M006325200 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 386BD UT WOS:000166039500112 PM 11018034 ER PT J AU Toukmaji, A Sagui, C Board, J Darden, T AF Toukmaji, A Sagui, C Board, J Darden, T TI Efficient particle-mesh Ewald based approach to fixed and induced dipolar interactions SO JOURNAL OF CHEMICAL PHYSICS LA English DT Article ID FAST MULTIPOLE METHOD; MOLECULAR-DYNAMICS; POLARIZABLE WATER; ELECTROSTATIC INTERACTIONS; CONSTANT-PRESSURE; SYSTEMS; SIMULATIONS; CHARGE; MODEL; SUMS AB We have implemented classical Ewald and particle-mesh Ewald (PME) based treatments of fixed and induced point dipoles into the sander molecular dynamics (MD) module of AMBER 6. During MD the induced dipoles can be propagated along with the atomic positions either by iteration to self-consistency at each time step, or by a Car-Parrinello (CP) technique using an extended Lagrangian formalism. In this paper we present the derivation of the new algorithms and compare the various options with respect to accuracy, efficiency, and effect on calculated properties of a polarizable water model. The use of PME for electrostatics of fixed charges and induced dipoles together with a CP treatment of dipole propagation in MD simulations leads to a cost overhead of only 33% above that of MD simulations using standard PME with fixed charges, allowing the study of polarizability in large macromolecular systems. (C) 2000 American Institute of Physics. [S0021- 9606(00)50547-9]. C1 Duke Univ, Dept Elect Engn, Durham, NC 27708 USA. NIEHS, Struct Biol Lab, Res Triangle Pk, NC 27709 USA. RP Toukmaji, A (reprint author), Conexant Syst Inc, Newport Beach, CA 92657 USA. NR 47 TC 210 Z9 210 U1 0 U2 38 PU AMER INST PHYSICS PI MELVILLE PA 2 HUNTINGTON QUADRANGLE, STE 1NO1, MELVILLE, NY 11747-4501 USA SN 0021-9606 J9 J CHEM PHYS JI J. Chem. Phys. PD DEC 22 PY 2000 VL 113 IS 24 BP 10913 EP 10927 DI 10.1063/1.1324708 PG 15 WC Chemistry, Physical; Physics, Atomic, Molecular & Chemical SC Chemistry; Physics GA 382TZ UT WOS:000165841300010 ER PT J AU Tian, GL Zhang, YB Zhang, TY Yang, FQ Ito, Y AF Tian, GL Zhang, YB Zhang, TY Yang, FQ Ito, Y TI Separation of tanshinones from Salvia miltiorrhiza Bunge by high-speed counter-current chromatography using stepwise elution SO JOURNAL OF CHROMATOGRAPHY A LA English DT Article DE Salvia miltiorrhiza; counter-current chromatography; plant materials; preparative chromatography; tanshinones ID PURIFICATION AB High-speed counter-current chromatography (HSCCC) was successfully used for isolation and purification of tanshinones from the roots of Salvia miltiorrhiza Bunge by stepwise elution. A set of three solvent systems and other experimental conditions were determined by analytical HSCCC. Using the optimized conditions, the preparative HSCCC separation was performed on 50 mg of crude light petroleum extract yielding pure tanshinones of tanshinone IIA (7 mg), tanshinone I (3 mg) and cryptotanshinone (4 mg) all at purities of over 95% in a single run. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NHLBI, Biophys Chem Lab, NIH, Bethesda, MD 20892 USA. Beijing Inst New Technol Applicat, Beijing 100035, Peoples R China. RP Ito, Y (reprint author), NHLBI, Biophys Chem Lab, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 7 TC 62 Z9 69 U1 1 U2 14 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0021-9673 J9 J CHROMATOGR A JI J. Chromatogr. A PD DEC 22 PY 2000 VL 904 IS 1 BP 107 EP 111 DI 10.1016/S0021-9673(00)00916-X PG 5 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 390QQ UT WOS:000166308200010 PM 11209896 ER PT J AU Feng, ZH Hong, JS Qi, QP Han, YF Wilson, B Iadarola, M Tiao, NY Bing, GY AF Feng, ZH Hong, JS Qi, QP Han, YF Wilson, B Iadarola, M Tiao, NY Bing, GY TI Cloning and expression of MP13 gene from rat hippocampus, a new factor related to guanosine triphosphate regulation SO NEUROSCIENCE LETTERS LA English DT Article DE Fos-related antigen; transcription factors; gene; cloning; guanosine triphosphate; immunoscreening; gel-shift ID TRANSCRIPTION FACTORS; INDUCTION; PROTEIN; COCAINE; RAB3A; BRAIN AB C-Fos and the Fos-related antigens (FRA) are induced by various stimuli. A novel 35-37 kDa FRA was induced much longer after the treatment using kainic acid (KA) and may be very important for neuronal survival after brain damage. To identify this long-term FRA, we have constructed a cDNA library derived from hippocampus after KA treatment and screened it with an antibody highly conserved M-peptide region of FRAs. One gene, MP13, was cloned with a 1662 bp open reading frame and coded for a 554-amino acid protein. MP13 has a leucine zipper region, a glutamine repeat region, and has high similarity to the activator of the small guanosine triphosphate (GTP)ase Rab5. Gel retardation analysis revealed that MP13 functions as a GTP regulation related factor. (C) 2000 Elsevier Science Ireland Ltd. AII rights reserved. C1 Univ Kentucky, Med Ctr, Dept Anat & Neurobiol, Lexington, KY 40536 USA. Univ Hong Kong, Queen Mary Hosp, Dept Med, Div Neurol, Hong Kong, Hong Kong, Peoples R China. NIEHS, Neuropharmacol Sect, Lab Pharmacol & Chem, NIH, Res Triangle Pk, NC 27709 USA. Hong Kong Univ Sci & Technol, Dept Biochem, Hong Kong, Hong Kong, Peoples R China. NIDR, Neurobiol & Anesthesiol Branch, NIH, Bethesda, MD 20892 USA. RP Bing, GY (reprint author), Univ Kentucky, Med Ctr, Dept Anat & Neurobiol, Lexington, KY 40536 USA. OI Han, Yifan/0000-0002-5833-069X; Bing, Guoying/0000-0003-0609-8152 NR 12 TC 0 Z9 1 U1 0 U2 0 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0304-3940 J9 NEUROSCI LETT JI Neurosci. Lett. PD DEC 22 PY 2000 VL 296 IS 2-3 BP 129 EP 132 DI 10.1016/S0304-3940(00)01641-4 PG 4 WC Neurosciences SC Neurosciences & Neurology GA 383AP UT WOS:000165859800017 PM 11108998 ER PT J AU Aguilera, G Rabadan-Diehl, C AF Aguilera, G Rabadan-Diehl, C TI Vasopressinergic regulation of the hypothalamic-pituitary-adrenal axis: implications for stress adaptation SO REGULATORY PEPTIDES LA English DT Article; Proceedings Paper CT 10th Annual Meeting of the European-Neuropeptide-Club CY MAY 10-13, 2000 CL INNSBRUCK, AUSTRIA SP European Neuropeptide Club DE hypothalamic paraventricular nucleus; vasopressin transcription; V1b receptor; V1b receptor mRNA; regulation ID CORTICOTROPIN-RELEASING HORMONE; RECEPTOR MESSENGER-RNA; ARGININE-VASOPRESSIN; GLUCOCORTICOID RECEPTOR; ADRENOCORTICOTROPIN SECRETION; PARAVENTRICULAR NUCLEUS; DIFFERENTIAL REGULATION; HETERONUCLEAR RNA; RIBONUCLEIC-ACID; MEDIAN-EMINENCE AB In addition to its role on water conservation, vasopressin (VP) regulates pituitary ACTH secretion by potentiating the stimulatory effects of corticotropin releasing hormone (CRH). The pituitary actions of VP are mediated by plasma membrane receptors of the V1b subtype, coupled to calcium-phospholipid signaling systems. VP is critical for adaptation of the hypothalamic-pituitary-adrenal (HPA) axis to stress as indicated by preferential expression of VP over CRH in parvocellular neurons of the hypothalamic paraventricular nucleus. and the upregulation of pituitary VP receptors during stress paradigms associated with corticotroph hyperresponsiveness. V1b receptor mRNA levels and coupling of the receptor to phospolipase C are stimulated by glucocorticoids, effects which may contribute to the refractoriness of VP-stimulated ACTH secretion to glucocorticoid feedback. The data suggest that vasopressinergic regulation of the HPA axis is critical for sustaining corticotroph responsiveness in the presence of high circulating glucocorticoid levels during chronic stress. (C) Published by Elsevier Science B.V. C1 NICHHD, Sect Endocrinol Physiol, Dev Endocrinol Branch, NIH, Bethesda, MD 20892 USA. RP Aguilera, G (reprint author), NICHHD, Sect Endocrinol Physiol, Dev Endocrinol Branch, NIH, Bldg 10,Rm 10N262,10 Ctr Dr MSC 1862, Bethesda, MD 20892 USA. NR 54 TC 226 Z9 235 U1 0 U2 10 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0167-0115 J9 REGUL PEPTIDES JI Regul. Pept. PD DEC 22 PY 2000 VL 96 IS 1-2 SI SI BP 23 EP 29 DI 10.1016/S0167-0115(00)00196-8 PG 7 WC Endocrinology & Metabolism; Physiology SC Endocrinology & Metabolism; Physiology GA 389PN UT WOS:000166247100005 PM 11102648 ER PT J AU Zemlyak, I Furman, S Brenneman, DE Gozes, I AF Zemlyak, I Furman, S Brenneman, DE Gozes, I TI A Novel peptide prevents death in enriched neuronal cultures SO REGULATORY PEPTIDES LA English DT Article; Proceedings Paper CT 10th Annual Meeting of the European-Neuropeptide-Club CY MAY 10-13, 2000 CL INNSBRUCK, AUSTRIA SP European Neuropeptide Club DE vasoactive intestinal peptide; activity-dependent neuroprotective protein activity-dependent neurotrophic factor; cerebral conical cultures ID VASOACTIVE-INTESTINAL-PEPTIDE; DEPENDENT NEUROTROPHIC FACTOR; ACTING NEUROPROTECTIVE PEPTIDE; MOLECULAR PATHOLOGY; ALZHEIMERS-DISEASE; GENE-EXPRESSION; FACTOR ADNF; CELL-DEATH; BRAIN; VIP AB We have recently cloned a novel protein (activity-dependent neuroprotective protein, ADNP) containing an 8-amino-acid, femtomolar-acting peptide, NAPVSIPQ (NAP). Here we show, for the first time, that NAP exerted a protective effect on glia-depleted neurons in culture. The number of surviving neurons was assessed in cerebral cortical cultures derived from newborn rats. In these cultures, a 24-h treatment with the beta -amyloid peptide (the Alzheimer's disease associated toxin) induced a 30-40% reduction in neuronal survival that was prevented by NAP (10(-13)-10(-11) M). Maximal survival was achieved at NAP concentrations of 10(-12) M. In a second set of experiments, a 5-day incubation period, with NAP added once (at the beginning of the incubation period) exhibited maximal protection at 10(-10) M NAP. In a third set of experiments, a 10-min period of glucose deprivation resulted in a 30-40% neuronal death that was prevented by a 24-h incubation with NAP. Glucose deprivation coupled with beta -amyloid treatment did not increase neuronal death, suggesting a common pathway. We thus conclude, that NAP can prevent neurotoxicity associated with direct action of the beta -amyloid peptide on neurons, perhaps through protection against impaired glucose metabolism. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Tel Aviv Univ, Sackler Fac Med, Dept Clin Biochem, IL-69978 Tel Aviv, Israel. NICHHD, Sect Dev & Mol Pharmacol, Dev Neurobiol Lab, NIH, Bethesda, MD 20892 USA. RP Gozes, I (reprint author), Tel Aviv Univ, Sackler Fac Med, Dept Clin Biochem, IL-69978 Tel Aviv, Israel. NR 31 TC 61 Z9 64 U1 0 U2 5 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0167-0115 J9 REGUL PEPTIDES JI Regul. Pept. PD DEC 22 PY 2000 VL 96 IS 1-2 SI SI BP 39 EP 43 DI 10.1016/S0167-0115(00)00198-1 PG 5 WC Endocrinology & Metabolism; Physiology SC Endocrinology & Metabolism; Physiology GA 389PN UT WOS:000166247100007 PM 11102650 ER PT J AU Veenstra, GJC Weeks, DL Wolffe, AP AF Veenstra, GJC Weeks, DL Wolffe, AP TI Distinct roles for TBP and TBP-like factor in early embryonic gene transcription in Xenopus SO SCIENCE LA English DT Article ID MAJOR DEVELOPMENTAL TRANSITION; BOX-BINDING PROTEIN; MIDBLASTULA TRANSITION; EXPRESSION; EMBRYOGENESIS; LAEVIS AB The TATA-binding protein (TBP) is believed to function as a key component of the general transcription machinery. We tested the role of TBP during the onset of embryonic transcription by antisense oligonucleotide-mediated turnover of maternal TBP messenger RNA. Embryos without detectable TBP initiated gastrulation but died before completing gastrulation. The expression of many genes transcribed by RNA polymerase II and III was reduced: however, some genes were transcribed with an efficiency identical to that of TBP-containing embryos. Using a similar antisense strategy, we found that the TBP-Like factor TLF/TRF2 is essential for development past the mid-blastula stage. Because TBP and a TLF factor play complementary roles in embryonic development, our results indicate that although similar mechanistic roles exist in common. TBP and TLF function differentially to control transcription of specific genes. C1 NICHHD, Mol Embryol Lab, Bethesda, MD 20892 USA. NICHHD, Mol Genet Lab, Bethesda, MD 20892 USA. Univ Iowa, Dept Biochem, Iowa City, IA 52242 USA. Sangamo Biosci, Richmond, CA 94804 USA. RP Veenstra, GJC (reprint author), NICHHD, Mol Embryol Lab, Bethesda, MD 20892 USA. RI Weeks, Daniel/F-6216-2010; Veenstra, Gert Jan C./D-4963-2012 OI Veenstra, Gert Jan C./0000-0002-7787-4940 NR 25 TC 96 Z9 99 U1 0 U2 4 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 USA SN 0036-8075 J9 SCIENCE JI Science PD DEC 22 PY 2000 VL 290 IS 5500 BP 2312 EP + DI 10.1126/science.290.5500.2312 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 385GQ UT WOS:000165995800047 PM 11125147 ER PT J AU Furey, ML Pietrini, P Haxby, JV AF Furey, ML Pietrini, P Haxby, JV TI Cholinergic enhancement and increased selectivity of perceptual processing during working memory SO SCIENCE LA English DT Article ID CEREBRAL BLOOD-FLOW; PREFRONTAL CORTEX; ATTENTION; SYSTEMS; TASK; ORGANIZATION; SCOPOLAMINE; DEMENTIA; DISEASE; TESTS AB Using functional magnetic resonance imaging, we investigated the mechanism by which cholinergic enhancement improves working memory. We studied the effect of the cholinesterase inhibitor physostigmine on subcomponents of this complex function. Cholinergic enhancement increased the selectivity of neural responses in extrastriate cortices during visual working memory, particularly during encoding, It also increased the participation of ventral extrastriate cortex during memory maintenance and decreased the participation of anterior prefrontal cortex. These results indicate that cholinergic enhancement improves memory performance by augmenting the selectivity of perceptual processing during encoding, thereby simplifying processing demands during memory maintenance and reducing the need for prefrontal participation. C1 NIMH, Lab Brain & Cognit, NIH, Bethesda, MD 20892 USA. Univ Pisa, Dept Human & Environm Sci, Pisa, Italy. RP Furey, ML (reprint author), NIMH, Lab Brain & Cognit, NIH, Bethesda, MD 20892 USA. RI Furey, Maura/H-5273-2013 NR 30 TC 212 Z9 215 U1 0 U2 2 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 USA SN 0036-8075 J9 SCIENCE JI Science PD DEC 22 PY 2000 VL 290 IS 5500 BP 2315 EP + DI 10.1126/science.290.5500.2315 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 385GQ UT WOS:000165995800048 PM 11125148 ER PT J AU De Vries, G Sherman, A AF De Vries, G Sherman, A TI Channel sharing in pancreatic beta-cells revisited: Enhancement of emergent bursting by noise SO JOURNAL OF THEORETICAL BIOLOGY LA English DT Article ID RESPIRATORY RHYTHM GENERATION; PRE-BOTZINGER COMPLEX; PACEMAKER NEURONS; COUPLED BURSTERS; MOUSE; OSCILLATIONS; MODEL; CLUSTERS; ISLET; HETEROGENEITY AB Secretion of insulin by electrically coupled populations of pancreatic beta -cells is governed by bursting electrical activity. Isolated beta -cells, however, exhibit atypical bursting or continuous spike activity. We study bursting as an emergent property of the population, focussing on interactions among the subclass of spiking cells. These are modelled by equipping the fast subsystem with a saddle-node-loop bifurcation, which makes it monostable. Such cells can only spike tonically or remain silent when isolated, but can be induced to burst with weak diffusive coupling. With stronger coupling, the cells revert to tonic spiking. We demonstrate that the addition of noise dramatically increases, via a phenomenon like stochastic resonance, the coupling range over which bursting is seen. (C) 2000 Academic Press. C1 Univ Alberta, Dept Math Sci, Edmonton, AB T6G 2G1, Canada. NIDDK, Math Res Branch, NIH, Bethesda, MD 20892 USA. RP De Vries, G (reprint author), Univ Alberta, Dept Math Sci, Edmonton, AB T6G 2G1, Canada. NR 34 TC 51 Z9 51 U1 0 U2 0 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-5193 J9 J THEOR BIOL JI J. Theor. Biol. PD DEC 21 PY 2000 VL 207 IS 4 BP 513 EP 530 DI 10.1006/jtbi.2000.2193 PG 18 WC Biology; Mathematical & Computational Biology SC Life Sciences & Biomedicine - Other Topics; Mathematical & Computational Biology GA 381UC UT WOS:000165781000005 PM 11093836 ER PT J AU Chen, L Chetkovich, DM Petralia, RS Sweeney, NT Kawasaki, Y Wenthold, RJ Bredt, DS Nicoll, RA AF Chen, L Chetkovich, DM Petralia, RS Sweeney, NT Kawasaki, Y Wenthold, RJ Bredt, DS Nicoll, RA TI Stargazin regulates synaptic targeting of AMPA receptors by two distinct mechanisms SO NATURE LA English DT Article ID MUTANT MOUSE; RAT-BRAIN; GLUTAMATE RECEPTORS; GAMMA-SUBUNIT; SYNAPSES; ORGANIZATION; EXPRESSION; PROTEINS; WAGGLER; FAMILY AB Stargazer, an ataxic and epileptic mutant mouse, lacks functional AMPA (alpha -amino-3-hydroxyl-5-methyl-4-isoxazolepropionate) receptors on cerebellar granule cells. Stargazin, the mutated protein, interacts with both AMPA receptor subunits and synaptic PDZ proteins, such as PSD-95. The interaction of stargazin with AMPA receptor subunits is essential for delivering functional receptors to the surface membrane of granule cells, whereas its binding with PSD-95 and related PDZ proteins through a carboxy-terminal PDZ-binding domain is required for targeting the AMPA receptor to synapses. Expression of a mutant stargazin lacking the PDZ-binding domain in hippocampal pyramidal cells disrupts synaptic AMPA receptors, indicating that stargazin-like mechanisms for targeting AMPA receptors may be widespread in the central nervous system. C1 Univ Calif San Francisco, Dept Mol & Cellular Pharmacol, San Francisco, CA 94143 USA. Univ Calif San Francisco, Dept Physiol, San Francisco, CA 94143 USA. Univ Calif San Francisco, Dept Neurol, San Francisco, CA 94143 USA. NIDCD, Neurochem Lab, NIH, Bethesda, MD 20892 USA. RP Nicoll, RA (reprint author), Univ Calif San Francisco, Dept Mol & Cellular Pharmacol, San Francisco, CA 94143 USA. NR 37 TC 655 Z9 675 U1 9 U2 43 PU MACMILLAN PUBLISHERS LTD PI LONDON PA PORTERS SOUTH, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD DEC 21 PY 2000 VL 408 IS 6815 BP 936 EP 943 PG 8 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 384NE UT WOS:000165951100039 PM 11140673 ER PT J AU Marx, SJ AF Marx, SJ TI Medical progress - Hyperparathyroid and hypoparathyroid disorders SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Review ID CHRONIC-RENAL-FAILURE; HORMONE-RELATED PEPTIDE; EXTRACELLULAR CA2+-SENSING RECEPTOR; MILD PRIMARY HYPERPARATHYROIDISM; CALCIUM-SENSING RECEPTOR; HUMAN PARATHYROID TUMORS; BONE-MINERAL DENSITY; VITAMIN-D-RECEPTOR; SECONDARY HYPERPARATHYROIDISM; POSTMENOPAUSAL WOMEN C1 NIDDKD, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. RP Marx, SJ (reprint author), NIDDKD, Metab Dis Branch, NIH, Bldg 10,Rm 9C-101,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 111 TC 248 Z9 264 U1 0 U2 7 PU MASSACHUSETTS MEDICAL SOC PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD DEC 21 PY 2000 VL 343 IS 25 BP 1863 EP 1875 DI 10.1056/NEJM200012213432508 PG 13 WC Medicine, General & Internal SC General & Internal Medicine GA 384KB UT WOS:000165940400008 PM 11117980 ER PT J AU Nabel, EG AF Nabel, EG TI Coronary heart disease in women. Reply SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Letter C1 NHLBI, Bethesda, MD 20892 USA. RP Nabel, EG (reprint author), NHLBI, Bldg 10, Bethesda, MD 20892 USA. NR 2 TC 0 Z9 0 U1 0 U2 0 PU MASSACHUSETTS MEDICAL SOC PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD DEC 21 PY 2000 VL 343 IS 25 BP 1894 EP 1894 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 384KB UT WOS:000165940400018 ER PT J AU Garbuglia, M Verzini, M Hofmann, A Huber, R Donato, R AF Garbuglia, M Verzini, M Hofmann, A Huber, R Donato, R TI S100A1 and S100B interactions with annexins SO BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH LA English DT Article DE S100A1; S100B; annexin V; annexin VI; calcium; interaction ID CALCIUM-BINDING PROTEINS; CALPACTIN-I; HEAD DOMAIN; 3-DIMENSIONAL STRUCTURE; CARDIOLIPIN VESICLES; ELECTRON-MICROSCOPY; CRYSTAL-STRUCTURE; DEPENDENT MANNER; S-100B PROTEIN; LIGHT-CHAIN AB Members of the annexin protein family interact with members of the S100 protein family thereby forming heterotetramers in which an S100 homodimer crossbridges two copies of the pertinent annexin. Previous work has shown that S100A1 and S100B bind annexin VI in a Ca2+-dependent manner and that annexin VI, but not annexin V, blocks the inhibitory effect of S100A1 and S100B on intermediate filament assembly. We show here that both halves of annexin VI (i.e., the N-terminal half or annexin VI-a and the C-terminal half or annexin VI-b) bind individual S100s on unique sites and that annexin VI-b, but not annexin VI-a, blocks the ability of S100A1 and S100B to inhibit intermediate filament assembly. We also show that the C-terminal extension of S100A1 (and, by analogy, S100B), that was previously demonstrated to be critical for S100A1 and S100B binding to several target proteins including intermediate filament subunits, is not part of the S100 surface implicated in the recognition of annexin VI, annexin VI-a, or annexin VI-b. Evaluation of functional properties with a liposome stability and a calcium influx assay reveals the ability of both S100 proteins to permeabilize the membrane bilayer in a similar fashion like annexins. When tested in combinations with different annexin proteins both S100 proteins mostly lead to a decrease in the calcium influx activity although not all annexin/S100 combinations behave in the same manner. Latter observation supports the hypothesis that the S100-annexin interactions differ mechanistically depending on the particular protein partners. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Univ Perugia, Dept Expt Med & Biochem Sci, Sect Anat, I-06122 Perugia, Italy. Max Planck Inst Biochem, Abt Strukturforsch, D-82152 Martinsried, Germany. NCI, Macromol Crystallog Lab, Program Struct Biol, FCRDC, Frederick, MD 21702 USA. RP Donato, R (reprint author), Univ Perugia, Dept Expt Med & Biochem Sci, Sect Anat, Via Giochetto CP 81 Succ 3, I-06122 Perugia, Italy. RI Hofmann, Andreas/B-9515-2008 OI Hofmann, Andreas/0000-0003-4408-5467 FU Telethon [922] NR 67 TC 25 Z9 25 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0167-4889 J9 BBA-MOL CELL RES JI Biochim. Biophys. Acta-Mol. Cell Res. PD DEC 20 PY 2000 VL 1498 IS 2-3 SI SI BP 192 EP 206 DI 10.1016/S0167-4889(00)00096-3 PG 15 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 387HP UT WOS:000166117400014 PM 11108963 ER PT J AU Schrag, D Cramer, LD Bach, PB Cohen, AM Warren, JL Begg, CB AF Schrag, D Cramer, LD Bach, PB Cohen, AM Warren, JL Begg, CB TI Influence of hospital procedure volume on outcomes following surgery for colon cancer SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID NEW-YORK-STATE; COLORECTAL-CANCER; PATIENT OUTCOMES; ADMINISTRATIVE DATA; MEDICARE PATIENTS; BREAST-CANCER; RECTAL-CANCER; HEALTH-CARE; MORTALITY; QUALITY AB Context Survival following high-risk cancer surgery, such as pancreatectomy and esophagectomy, is superior at hospitals where high volumes of these procedures are per formed. Conflicting evidence exists as to whether the association between hospital experience and favorable health outcomes also applies to more frequently performed operations, such as those for colon cancer. Objective To determine whether hospital procedure volume predicts survival following colon cancer surgery. Design, Setting, and Participants Retrospective cohort study of data from the Surveillance, Epidemiology and End Results-Medicare linked database on 27986 colon cancer patients aged 65 years and older who had surgical resection for primary adenocarcinoma diagnosed between 1991 and 1996. Main Outcome Measures Thirty-day postoperative mortality and overall and cancer-specific long-term survival, by hospital procedure volume. Results We found small differences in 30-day postoperative mortality for patients treated at low- vs high-volume hospitals (3.5% at hospitals in the top-volume quartile vs 5.5% at hospitals in the bottom-volume quartile). However, the correlation was statistically significant and persisted after adjusting for age at diagnosis, sex, race, cancer stage, comorbid illness, socioeconomic status, and acuity of hospitalization (P<.001). The association was evident for subgroups with stage I, II, and III disease. Hospital Volume directly correlated with survival beyond 30 days and also was not attributable to differences in case mix (P<.001). The association between hospital volume and long-term survival was concentrated among patients with stage II and III disease (P<.001 for both). Among stage III patients. Variation in use of adjuvant chemotherapy did not explain this finding. Conclusion Our data suggest that hospital procedure volume predicts clinical outcomes following surgery for colon cancer, although the absolute magnitudes of these differences are modest in comparison with the variation observed for higher-risk cancer surgeries. C1 Mem Sloan Kettering Canc Ctr, Dept Epidemiol & Biostat, Hlth Outcomes Res Grp, New York, NY 10021 USA. Mem Sloan Kettering Canc Ctr, Dept Surg, Hlth Outcomes Res Grp, New York, NY 10021 USA. NCI, Appl Res Branch, Bethesda, MD 20892 USA. RP Schrag, D (reprint author), Mem Sloan Kettering Canc Ctr, Dept Epidemiol & Biostat, Hlth Outcomes Res Grp, 1275 York Ave, New York, NY 10021 USA. NR 38 TC 331 Z9 336 U1 1 U2 6 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD DEC 20 PY 2000 VL 284 IS 23 BP 3028 EP 3035 DI 10.1001/jama.284.23.3028 PG 8 WC Medicine, General & Internal SC General & Internal Medicine GA 382VC UT WOS:000165847200028 PM 11122590 ER PT J AU Wang, PY Brank, AS Banavali, NK Nicklaus, MC Marquez, VE Christman, JK MacKerell, AD AF Wang, PY Brank, AS Banavali, NK Nicklaus, MC Marquez, VE Christman, JK MacKerell, AD TI Use of oligodeoxyribonucleotides with conformationally constrained abasic sugar targets to probe the mechanism of base flipping by HhaI DNA (cytosine C5)-methyltransferase SO JOURNAL OF THE AMERICAN CHEMICAL SOCIETY LA English DT Article ID BICYCLO<3.1.0>HEXANE TEMPLATE; MOLECULAR-DYNAMICS; CRYSTAL-STRUCTURE; CARBOCYCLIC NUCLEOSIDES; PSEUDOROTATIONAL CYCLE; REVERSE-TRANSCRIPTASE; NEPLANOCIN-C; METHYLTRANSFERASE; GLYCOSYLASE; ANALOGS AB X-ray crystallographic studies of HhaI DNA (cytosine-C5)-methyltransferase (M.HhaI) covalently linked to methylated 5-fluorocytosine in DNA provided the first direct evidence that the cytosine residue targeted for methylation was "flipped" out of the helix during the transfer reaction. Subsequent studies indicated that removal of the target cytosine base, i.e., introduction of an abasic site, enhanced binding of M.HhaI to DNA and that the conformation of the sugar-phosphate backbone at the abasic site in the resultant complexes was the same as that of the sugar attached to a "flipped" cytosine. In the present study, pseudorotationally constrained sugar analogues, based on bicyclo[3.1.0]hexane templates, were placed in DNA duplexes as abasic target sites in the M.HhaI recognition sequence. Biochemical studies demonstrate that binding affinity of M.HhaI for abasic sites increases when the abasic target sugar analogue is constrained to the south conformation and decreases when it is constrained to the north conformation. In native gel-shift assays, M.HhaI exhibits a "closed" conformation when bound to the abasic south or abasic furanose analogues, whereas an "open" conformation predominates with the abasic north analogue. A structural understanding of these results was obtained via molecular dynamics simulations of the DNA duplex alone and in ternary complex with M.HhaI and cofactor, along with quantum mechanical calculations on model compounds representative of the abasic and modified sugars. Binding affinities are shown to be related to the ability of the abasic sugar analogues to spontaneously flip out of the DNA duplex. Enhanced binding of the abasic south analogue is suggested to be due to its increased capacity for sampling the experimentally observed conformation of the DNA target site in the M.HhaI ternary complex. Decreased binding of the north analogue is due to decreased flexibility of the phosphodiester backbone associated with a north pseudorotation angle, thereby inhibiting spontaneous flipping of the sugar moiety out of the DNA duplex. Spontaneous flipping of the sugar moiety out of the DNA duplex is also suggested to facilitate formation of a "closed" complex between M.HhaI and DNA whereas partial or no flipping favors the "open" conformation. These results show that introduction of structural constraints into DNA that induce enhanced sampling of protein-bound conformations facilitate DNA-protein binding. Implications of the present results with respect to the mechanism of base flipping in the M.HhaI catalytic cycle are discussed. C1 NCI, Med Chem Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. Univ Nebraska, Med Ctr 984525, Dept Biochem & Mol Biol, Omaha, NE 68198 USA. Univ Nebraska, Med Ctr 984525, Eppley Canc Ctr, Omaha, NE 68198 USA. Univ Maryland, Sch Med, Dept Pharmaceut Sci, Baltimore, MD 21201 USA. RP Marquez, VE (reprint author), NCI, Med Chem Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RI Nicklaus, Marc/N-4183-2014; OI Banavali, Nilesh/0000-0003-2206-7049; MacKerell, Alex/0000-0001-8287-6804 NR 67 TC 47 Z9 48 U1 0 U2 6 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0002-7863 J9 J AM CHEM SOC JI J. Am. Chem. Soc. PD DEC 20 PY 2000 VL 122 IS 50 BP 12422 EP 12434 DI 10.1021/ja001989s PG 13 WC Chemistry, Multidisciplinary SC Chemistry GA 387ZZ UT WOS:000166154100002 ER PT J AU Paik, S Bryant, J Tan-Chiu, E Yothers, G Park, C Wickerham, DL Wolmark, N AF Paik, S Bryant, J Tan-Chiu, E Yothers, G Park, C Wickerham, DL Wolmark, N TI HER2 and choice of adjuvant chemotherapy for invasive breast cancer: National Surgical Adjuvant Breast and Bowel Project Protocol B-15 SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Article ID THERAPY; ERBB-2; DOXORUBICIN; WOMEN AB Background: Recent retrospective analyses have suggested that breast cancer patients whose tumors overexpress HER2 derive preferential benefit from treatment with anthracyclines such as doxorubicin. This has led some clinicians to propose that HER2 should be used as a predictive marker in choosing between anthracycline-based regimens and combination chemotherapy with cyclophosphamide, methotrexate, and 5-fluorouracil (CMF). We evaluated this recommendation in a retrospective study of National Surgical Adjuvant Breast and Bowel Project Protocol B-15, in which patients received a combination of doxorubicin and cyclophosphamide (AC), CMF, or AC followed by CMF, We hypothesized that AC would be superior to CMF only in the HER2-positive patients. Methods: Immunohistochemical detection of HER2 was performed on tumor sections from 2034 of 2295 eligible patients. We used statistical analysis to evaluate the interaction between the efficacy of the assigned treatments and HER2 overexpression. All statistical tests were two-sided. Results: Tumor sections from 599 patients (29%) stained positive for HER2. AC was superior to CMF in HER2-positive patients only, although differences in outcomes did not reach statistical significance. In the HER2-positive cohort, relative risks of failure (i.e., after AC treatment as compared with CMF treatment) were 0.84 for disease-free survival (DFS) (95% confidence interval [CI] = 0.65-1.07; P = .15), 0.82 for survival (95% CT = 0.63-1.06; p = .14), and 0.80 for recurrence-free survival (RFS) (95% CI = 0.62-1.04; P = .10). Tests for interaction between treatment and HER2 status were suggestive but not statistically significant (P = .19 for DFS, P = .11 for survival, and P = .08 for RFS), Conclusions: These results, together with overview results indicating minor overall superiority for anthracycline-based regimens relative to CMF, indicate a preference for the AC regimen in patients with HER2-positive tumors. Both AC and CMF regimens may be considered for patients with HER2-negative tumors. C1 Allegheny Gen Hosp, Natl Surg Adjuvant Breast & Bowel Project, Div Pathol, Pittsburgh, PA 15212 USA. Univ Pittsburgh, NSABP, Pittsburgh, PA 15260 USA. Univ Pittsburgh, Dept Stat, Pittsburgh, PA 15260 USA. Univ Pittsburgh, Dept Biostat, Pittsburgh, PA 15260 USA. NSABP Operat Ctr, Pittsburgh, PA USA. RP Paik, S (reprint author), Natl Surg Adjuvant Breast & Bowel Project, Div Pathol, E Commons Profess Bldg,4 Allegheny Ctr,5th Floor, Pittsburgh, PA 15212 USA. OI Yothers, Greg/0000-0002-7965-7333 FU NCI NIH HHS [U10CA12027, U10CA37377, U10CA69651, U10CA69974] NR 14 TC 202 Z9 207 U1 0 U2 1 PU NATL CANCER INSTITUTE PI BETHESDA PA 9030 OLD GEORGETOWN RD, BETHESDA, MD 20814 USA SN 0027-8874 J9 J NATL CANCER I JI J. Natl. Cancer Inst. PD DEC 20 PY 2000 VL 92 IS 24 BP 1991 EP 1998 DI 10.1093/jnci/92.24.1991 PG 8 WC Oncology SC Oncology GA 384HM UT WOS:000165936800010 PM 11121461 ER PT J AU Kleinerman, RA Tarone, RE Abramson, DH Seddon, JM Li, FP Tucker, MA AF Kleinerman, RA Tarone, RE Abramson, DH Seddon, JM Li, FP Tucker, MA TI Hereditary retinoblastoma and risk of lung cancer SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Article ID 2ND NONOCULAR TUMORS; BILATERAL RETINOBLASTOMA; RELATIVES; SURVIVORS; MORTALITY; EXPRESSION; CARCINOMA; NEOPLASMS; EXCESS; DEATHS C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. New York Presbyterian Hosp, Cornell Med Ctr, New York, NY USA. Massachusetts Eye & Ear Infirm, Boston, MA 02114 USA. Dana Farber Canc Inst, Div Canc Epidemiol & Control, Boston, MA 02115 USA. Harvard Univ, Sch Publ Hlth, Boston, MA 02115 USA. RP Kleinerman, RA (reprint author), NIH, Execut Plaza S,Rm 7044, Bethesda, MD 20892 USA. RI Tucker, Margaret/B-4297-2015; OI Kleinerman, Ruth/0000-0001-7415-2478 FU NCI NIH HHS [N01CP81121] NR 30 TC 39 Z9 39 U1 0 U2 2 PU NATL CANCER INSTITUTE PI BETHESDA PA 9030 OLD GEORGETOWN RD, BETHESDA, MD 20814 USA SN 0027-8874 J9 J NATL CANCER I JI J. Natl. Cancer Inst. PD DEC 20 PY 2000 VL 92 IS 24 BP 2037 EP 2039 DI 10.1093/jnci/92.24.2037 PG 3 WC Oncology SC Oncology GA 384HM UT WOS:000165936800016 PM 11121467 ER PT J AU Koch, WH de Henestrosa, ARF Woodgate, R AF Koch, WH de Henestrosa, ARF Woodgate, R TI Identification of mucAB-like homologs on two IncT plasmids, R394 and Rts-1 SO MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS LA English DT Article DE SOS mutagenesis; UmuC; Tus; DinI; LexA; C. freundii ID DIFFERENT INCOMPATIBILITY GROUPS; 60-MEGADALTON CRYPTIC PLASMID; DNA-POLYMERASE-ETA; ESCHERICHIA-COLI; SALMONELLA-TYPHIMURIUM; INDUCIBLE MUTAGENESIS; UV MUTAGENESIS; UMU OPERON; ULTRAVIOLET-LIGHT; SEQUENCE-ANALYSIS AB Recent phylogenetic analysis of the superfamily of lesion-replicating DNA polymerases suggest that they can be broadly divided into four sub-groups comprised of UmuC-like, DinB-like, Rev1-like and Rad30-like proteins. The UmuC-like sub-family is best characterized at the genetic level and sequence analysis of eleven umu orthologs, residing on bacterial chromosomes or on self-transmissible R-plasmids allows further subdivision into five sub-groups (UmuDC, MucAB, ImpAB, RumAB and RulAB) based on amino acid sequence conservation. Some of these orthologs are apparently inactive in situ, but may promote increased mutagenesis and survival when subcloned and expressed from high-copy number plasmids. We were, therefore, interested in devising an assay that would identify umuC-like genes in situ in the absence of a functional assay. To this end, degenerate primers directed towards conserved amino acid regions within the UmuC-like sub-family of DNA polymerases were designed and used to identify mucAB-like operons on the IncT plasmids, R394 and Rts-1. Interestingly, DNA sequence analysis of an similar to7 kb region of R394 identified two LexA-regulated genes immediately downstream of mucAB((R394)) that are similar to the chromosomally-encoded Escherichia coli tus gene and the IncI plasmid-encoded impC gene, respectively. Analysis of the R394 and Rts-1 mucB genes revealed that both contain insertions which result in the expression of a truncated inactive MucB protein. While R394 was unable to restore mutagenesis functions to a Delta umuDC E. coli strain, Rts-1 surprisingly promoted significant levels of MMS-induced SOS mutagenesis, raising the possibility that Rts-1 encodes another, yet unidentified, umu-like homolog. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NICHHD, Sect DNA Replicat Repair & Mutagenesis, NIH, Bethesda, MD 20892 USA. US FDA, Mol Biol Branch, Washington, DC 20204 USA. RP Woodgate, R (reprint author), NICHHD, Sect DNA Replicat Repair & Mutagenesis, NIH, Bldg 6,Room 1A13,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 63 TC 9 Z9 10 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0027-5107 J9 MUTAT RES-FUND MOL M JI Mutat. Res.-Fundam. Mol. Mech. Mutagen. PD DEC 20 PY 2000 VL 457 IS 1-2 BP 1 EP 13 DI 10.1016/S0027-5107(00)00134-2 PG 13 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology SC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology GA 385CB UT WOS:000165983100001 PM 11106794 ER PT J AU Erexson, GL Tindall, KR AF Erexson, GL Tindall, KR TI Micronuclei and gene mutations in transgenic Big Blue (R) mouse and rat fibroblasts after exposure to the epoxide metabolites of 1,3-butadiene SO MUTATION RESEARCH-GENETIC TOXICOLOGY AND ENVIRONMENTAL MUTAGENESIS LA English DT Article DE 1,3-butadiene; butadiene monoepoxide; butadiene diolepoxide; diepoxybutane; micronuclei; mutations ID SISTER-CHROMATID EXCHANGES; IN-VIVO; SPECIES-DIFFERENCES; BUTADIENE METABOLITES; DNA-ADDUCTS; INHALATION EXPOSURES; HUMAN-LYMPHOCYTES; MICE; DIEPOXYBUTANE; VITRO AB 1,3-Butadiene (BD) is a commodity compound and by-product in the manufacture of synthetic rubber that elicits a differential carcinogenic response in rodents after chronic exposure. Mice are up to approximately 1000-fold more sensitive to the tumorigenicity of inhaled ED than rats, thereby confounding human risk assessment analyses. Rodent transgenic in vivo and in vitro models have been recently utilized for generating genetic toxicology data in support of risk assessment studies. However, studies have not been extended to investigate multiple endpoints of genetic damage using in vitro transgenic models. The goal of this study was to evaluate possible differences in the production of genetic damage in transgenic Big Blue(R) mouse IBBM1) and rat (BBR1) fibroblasts exposed to three predominant epoxide metabolites of ED. Analyses of cytotoxicity, micronucleus (MN) formation, cll mutant frequency (MF) and apoptosis were assessed after in vitro exposure of BBM1 and BBR1 cells exposed to various concentrations of butadiene monoepoxide (BMO), diepoxybutane (DEB) and butadiene diolepoxide (BDE). Both BMO and DEB reduced cell survival in BBM1 and BBR1 cells. However, BDE decreased cell survival only in BBM1 cells at the concentrations evaluated. Concentration-dependent increases in the formation of MN was observed in both BBM1 and BBR1 cells, with DEB being the most potent followed by BDE and then BMO. The dose-response for mutations induced at the cll locus was essentially equal after DEB exposure of BBM 1 and BBR1 fibroblasts. In contrast, the cll MF was significantly increased only in BBM1 cells after exposure to either BMO or BDE. These data demonstrate a differential genetic response for gene mutations but not for MN formation in transgenic BBM1 and BBR1 fibroblasts and suggest a rodent species-specific difference in the persistence of DNA damage that results in gene mutations. In addition, apoptosis was observed in BBR1 cells but not in BBM1 cells when treated with any of the three ED epoxide metabolites. This response may partially explain the differential response to mutations induced by BMO and BDE. These data offer insight into specific differences in mouse and rat cells with respect to their response to ED epoxide metabolites. Such data may help to explain the different tumorigenicity results observed in rodent ED carcinogenicity studies. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIEHS, Lab Environm Carcinogenesis & Mutagenesis, Mol Mutagenesis Grp, Res Triangle Pk, NC 27709 USA. RP Erexson, GL (reprint author), Covance Labs Inc, Dept Genet & Mol Toxicol, 9200 Leesburg Pike, Vienna, VA 22182 USA. NR 49 TC 9 Z9 10 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 1383-5718 J9 MUTAT RES-GEN TOX EN JI Mutat. Res. Genet. Toxicol. Environ. Mutagen. PD DEC 20 PY 2000 VL 472 IS 1-2 BP 105 EP 117 DI 10.1016/S1383-5718(00)00136-4 PG 13 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology SC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology GA 385WW UT WOS:000166029000010 PM 11113703 ER PT J AU Witt, KL Zeiger, E Tice, RR van Birgelen, APJM AF Witt, KL Zeiger, E Tice, RR van Birgelen, APJM TI The genetic toxicity of 3,3 ',4,4 '-tetrachloroazobenzene and 3,3 ',4,4 '-tetrachloroazoxybenzene: discordance between acute mouse bone marrow and subchronic mouse peripheral blood micronucleus test results SO MUTATION RESEARCH-GENETIC TOXICOLOGY AND ENVIRONMENTAL MUTAGENESIS LA English DT Article DE micronuclei; genotoxicity; clastogenicity; NCE; PCE ID MICE; PHENOLPHTHALEIN; ERYTHROCYTES; INDUCTION; RATS; SALICYLAZOSULFAPYRIDINE; SENSITIVITY; EXPOSURE; INDUCERS; PROTOCOL AB 3,3',4,4'-Tetrachloroazobenzene (TCAB) and 3,3',4,4'-tetrachloroazoxybenzene (TCAOB) are dioxin-like chemicals that were investigated for toxicity in 13-week gavage studies in male and female B6C3F(1) mice and F344N rats by the National Toxicology Program. As part of the comprehensive toxicological investigation of these chemicals, peripheral blood smears from mice treated 5 days per week for 13 weeks with 0.1-30 mg/kg/day TCAB or TCAOB were analyzed for the frequency of micronucleated (MN) normochromatic erythrocytes (NCE). Both chemicals produced significant increases in MN-NCE in male and female mice. In contrast to these positive results in subchronic exposure studies, no significant increases were seen in acute bone marrow MN tests in male mice administered three daily injections of 50-200 mg/kg/day TCAB and TCAOB. The results with TCAB and TCAOB suggest that the routine integration of MN rests with subchronic toxicity studies may allow detection of mutagenic activity for some chemicals that fail to elicit responses in short-term, high dose tests. In addition, the integration of mutagenicity tests into general toxicity tests reduces the use of laboratory animals and the cost of the testing. (C) 2000 Published by Elsevier Science B.V. C1 ILS Inc, Res Triangle Pk, NC 27709 USA. NIEHS, Res Triangle Pk, NC 27709 USA. RP Witt, KL (reprint author), ILS Inc, POB 13501, Res Triangle Pk, NC 27709 USA. NR 33 TC 9 Z9 9 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 1383-5718 J9 MUTAT RES-GEN TOX EN JI Mutat. Res. Genet. Toxicol. Environ. Mutagen. PD DEC 20 PY 2000 VL 472 IS 1-2 BP 147 EP 154 DI 10.1016/S1383-5718(00)00143-1 PG 8 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology SC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology GA 385WW UT WOS:000166029000014 PM 11113707 ER PT J AU Roberts-Thomson, SJ Snyderwine, EG AF Roberts-Thomson, SJ Snyderwine, EG TI Characterization of peroxisome proliferator-activated receptor alpha in normal rat mammary gland and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine-induced mammary gland tumors from rats fed high and low fat diets SO TOXICOLOGY LETTERS LA English DT Article DE PPAR alpha; dietary fat; PhIP; rat; mammary gland; tumor ID SPRAGUE-DAWLEY RATS; HUMAN BREAST-CANCER; POLYMERASE CHAIN-REACTION; PPAR-ALPHA; METABOLIC-ACTIVATION; ADDUCT FORMATION; ACIDS ACTIVATE; RETINOIC ACID; MESSENGER-RNA; IN-VITRO AB Normal Sprague-Dau ley rat mammary gland epithelial cells and mammary gland carcinomas induced by 2-amino-1 -methyl-6-phenylimidazo[4,5-b]pyridine, a carcinogen found in the diet, were examined for the expression of peroxisome proliferator-activated receptor alpha (PPAR alpha). PPAR alpha mRNA and protein was detected in normal and tumor tissue by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry. By quantitative RT-PCR, carcinomas had a 12-fold higher expression than control mammary glands, a statistically significant difference. PPAR alpha expression was examined in carcinomas and normal tissues from rats on high fat (23.5/% corn oil) and low fat (5% corn oil) diets. Although neither carcinomas, nor control tissues showed statistically significant differences between the two diet groups, PPAR alpha expression was the highest in carcinomas from rats on the high fat diet. The expression of PPAR alpha in normal mammary gland and its significant elevation in mammary gland carcinomas raises the possibility of its involvement in mammary gland physiology and pathophysiology. (C) 2000 Published by Elsevier Science Ireland Ltd. All rights reserved. C1 Univ Queensland, Sch Pharm, St Lucia, Qld 4072, Australia. NIH, Chem Carcinogenesis Sect, Expt Carcinogenesis Lab, DBS, Bethesda, MD 20892 USA. RP Roberts-Thomson, SJ (reprint author), Univ Queensland, Sch Pharm, St Lucia, Qld 4072, Australia. RI Roberts-Thomson, Sarah/B-4282-2011 OI Roberts-Thomson, Sarah/0000-0001-8202-5786 NR 46 TC 17 Z9 17 U1 0 U2 0 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0378-4274 J9 TOXICOL LETT JI Toxicol. Lett. PD DEC 20 PY 2000 VL 118 IS 1-2 BP 79 EP 86 DI 10.1016/S0378-4274(00)00265-4 PG 8 WC Toxicology SC Toxicology GA 387UU UT WOS:000166140900010 PM 11137312 ER PT J AU Moriuchi, H Moriuchi, M AF Moriuchi, H Moriuchi, M TI In vitro induction of HIV-1 replication in resting CD4(+) T cells derived from individuals with undetectable plasma viremia upon stimulation with human T-cell leukemia virus type I SO VIROLOGY LA English DT Article; Proceedings Paper CT 6th Conference on Retroviruses and Opportunistic Infections CY JAN 31-FEB 04, 1999 CL CHICAGO, ILLINOIS ID HUMAN-IMMUNODEFICIENCY-VIRUS; ACTIVE ANTIRETROVIRAL THERAPY; NF-KAPPA-B; HTLV-I; HUMAN CYTOMEGALOVIRUS; VIRAL LOAD; INFECTION; COINFECTION; CXCR4; VIVO AB Microbial coinfections have been associated with transient bursts of human immunodeficiency virus (HIV) viremia in patients. In this study we investigated whether human T-cell leukemia virus type I (HTLV-I), another human retrovirus that is prevalent among certain HIV-infected populations, can induce HIV-1 replication in patients who had been successfully treated with highly active antiretroviral therapy. We demonstrate that supernatants from HTLV-l-producing MT-2 cells can induce in vitro replication of HIV-I from highly purified, resting CD4(+) T cells obtained from individuals with undetectable plasma viremia. Depletion of proinflammatory cytokines from the supernatants reduced, but did not abrogate, the ability to induce HIV-1 replication, indicating that other factors such as HTLV-I Tax or Env also have a role. The HTLV-l-mediated effect does not require productive infection: exposure to heat-inactivated HTLV-I virions, purified Tax protein, or HTLV-I Env glycoprotein also induced expression of HIV-I. Furthermore, we demonstrate that coculture of resting CD4(+) T cells with autologous CD8(+) T cells markedly inhibits the HTLV-l-induced virus replication. Our results suggest that coinfection with HTLV-I may induce viral replication in the latent viral reservoirs; however, CD8(+) T cells may play an important role in controlling the spread of virus upon microbial stimulation. (C) 2000 Academic Press. C1 NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. RP Moriuchi, H (reprint author), Nagasaki Univ, Sch Med, Dept Pediat, 1-7-1 Sakamoto, Nagasaki 8528501, Japan. NR 33 TC 7 Z9 7 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0042-6822 J9 VIROLOGY JI Virology PD DEC 20 PY 2000 VL 278 IS 2 BP 514 EP 519 DI 10.1006/viro.2000.0684 PG 6 WC Virology SC Virology GA 388EK UT WOS:000166164300020 PM 11118373 ER PT J AU Moriuchi, M Moriuchi, H Williams, R Straus, SE AF Moriuchi, M Moriuchi, H Williams, R Straus, SE TI Herpes simplex virus infection induces replication of human immunodeficiency virus type 1 SO VIROLOGY LA English DT Article DE human immunodeficiency virus type 1; herpes simplex virus; macrophages; NF-kappa B; glycoproteins; proinflammatory cytokines ID GLYCOPROTEIN-D; T-CELLS; HIV; ENTRY; CYTOMEGALOVIRUS; TRANSCRIPTION; TRANSMISSION; LOCALIZATION; INDUCTION; PROTEIN AB Genital herpes has been associated with increased efficiency of the sexual transmission and enhanced replication of human immunodeficiency virus type 1 (HIV-1). In this study we demonstrate that exposure to infectious or heat-inactivated herpes simplex virus (HSV) type 1 or 2 virions increases HIV-I expression in macrophages at least in part by inducing NF-kappaB activity. Neutralizing antibodies to the HSV glycoprotein gB or go markedly attenuated these virion-mediated effects on HIV-1 expression in macrophages. Thus HSV infection of macrophages that reside in genital mucosal tissue induces HIV-1 replication in these cells. Our study may have implications for the management of patients who are coinfected with the two viruses, (C) 2000 Academic Press. C1 Nagasaki Univ, Grad Sch Med Sci, Dept Mol Microbiol & Immunol, Div Med Virol, Nagasaki 8528501, Japan. NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. NIAID, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. RP Moriuchi, M (reprint author), Nagasaki Univ, Grad Sch Med Sci, Dept Mol Microbiol & Immunol, Div Med Virol, Nagasaki 8528501, Japan. NR 28 TC 58 Z9 58 U1 1 U2 1 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0042-6822 J9 VIROLOGY JI Virology PD DEC 20 PY 2000 VL 278 IS 2 BP 534 EP 540 DI 10.1006/viro.2000.0667 PG 7 WC Virology SC Virology GA 388EK UT WOS:000166164300022 PM 11118375 ER PT J AU Amin, S Zhang, YQ Sawin, DT Evans, SR Hannan, MT Kiel, DP Wilson, PWF Felson, DT AF Amin, S Zhang, YQ Sawin, DT Evans, SR Hannan, MT Kiel, DP Wilson, PWF Felson, DT TI Association of hypogonadism and estradiol levels with bone mineral density in elderly men from the framingham study SO ANNALS OF INTERNAL MEDICINE LA English DT Article ID IDIOPATHIC HYPOGONADOTROPIC HYPOGONADISM; SEX STEROID-LEVELS; AROMATASE DEFICIENCY; OSTEOPOROTIC FRACTURES; PHYSICAL-ACTIVITY; ESTROGEN THERAPY; TESTICULAR AXIS; HIP-FRACTURES; RISK-FACTORS; HEALTHY-MEN AB Background: Both hypogonadism and low estrogen levels adversely affect bone health in young men. In elderly men, who are at greatest risk for osteoporotic fracture, the influence of hypogonadism on bone mineral density remains unclear, as does the relative effect of estrogen status compared to hypogonadism. Objective: To examine the relation of hypogonadism and estrogen status to bone mineral density in elderly men. Design: Community-based, prospective cohort study. Setting: Framingham, Massachusetts. Patients: Male participants of the Framingham Study. Measurements: Total testosterone, total estradiol, and luteinizing hormone were measured in participants at all four biennial examinations from 1981 to 1989. Values from at least three of four examinations were averaged. Hypogonadism was defined as a mean testosterone level less than 10.4 nmol/L (<3.0 ng/mL) or a mean luteinizing hormone level of 20 IU/L or greater. An alternate definition of hypogonadism based only on a mean testosterone level less than 10.4 nmol/L (<3.0 ng/mL) was also used. In 1988-1989, bone mineral density was measured at the proximal femur (femoral neck, Ward triangle, and trochanter) and lumbar spine by using dual-photon absorptiometry and at the radial shaft by using single-photon absorptiometry. The association of hypogonadism with bone mineral density was examined with adjustment for confounders, including estradiol levels. A similar model that adjusted for hypogonadism was used to examine the association of estradiol level (ranked as quartiles) with bone mineral density. Results: of 448 men with bone mineral density measurements, 405 had evaluable hormone levels (mean age, 75.7 years [range, 68 to 96 years]); 71 (17.5%) of the 405 men were hypogonadal. Bone mineral density at any site did not significantly differ in hypogonadal men compared with eugonadal men (for example, bone mineral density at the femoral neck was 0.89 g/cm(2) vs. 0.87 g/cm(2), respectively; P > 0.2), even when alternate definitions of hypogonadism were used. In contrast, compared with the lowest estradiol quartile, men with higher estradiol levels had greater mean bone mineral density at all sites (for example, bone mineral density at the femoral neck was 0.84 g/cm(2), 0.88 g/cm(2), 0.86 g/cm(2), and 0.91 g/cm(2) from the lowest to the highest estradiol quartile; P for trend = 0.002). The difference in mean bone mineral density between men in the lowest and those in the highest estradiol quartile levels was similar to the effect of 10 years of aging on bone mineral density. Conclusions: In elderly men, hypogonadism related to aging has little influence on bone mineral density, but serum estradiol levels have a strong and positive association with bone mineral density. C1 Boston Univ, Sch Med, Multipurpose Arthrit & Musculoskeletal Dis Ctr, Boston, MA 02118 USA. Hebrew Rehabil Ctr Aged, Res & Training Inst, Boston, MA 02131 USA. Harvard Univ, Sch Med, Div Aging, Boston, MA 02115 USA. Vet Adm Headquarters, Washington, DC USA. NHLBI, Framingham Heart Study, Framingham, MA USA. RP Amin, S (reprint author), Mayo Clin, Div Rheumatol, 200 1st St SW, Rochester, MN 55905 USA. OI Kiel, Douglas/0000-0001-8474-0310 FU NHLBI NIH HHS [N01-HC-38038]; NIAMS NIH HHS [AR/AG41398, AR20613] NR 65 TC 179 Z9 181 U1 1 U2 266 PU AMER COLL PHYSICIANS PI PHILADELPHIA PA INDEPENDENCE MALL WEST 6TH AND RACE ST, PHILADELPHIA, PA 19106-1572 USA SN 0003-4819 J9 ANN INTERN MED JI Ann. Intern. Med. PD DEC 19 PY 2000 VL 133 IS 12 BP 951 EP 963 PG 13 WC Medicine, General & Internal SC General & Internal Medicine GA 384NT UT WOS:000165952300003 PM 11119396 ER PT J AU Djousse, L Levy, D Murabito, JM Cupples, LA Ellison, RC AF Djousse, L Levy, D Murabito, JM Cupples, LA Ellison, RC TI Alcohol consumption and risk of intermittent claudication in the Framingham Heart Study SO CIRCULATION LA English DT Article DE alcohol; smoking; peripheral vascular disease ID PERIPHERAL ARTERIAL-DISEASE; DENSITY-LIPOPROTEIN CHOLESTEROL; EDINBURGH-ARTERY; CARDIOVASCULAR-DISEASE; GENERAL-POPULATION; OCCLUSIVE DISEASE; MORTALITY; WINE; PREVALENCE; DINNER AB Background-Intermittent claudication (IC) is associated with an increased risk of cardiovascular disease morbidity and mortality. The relation of alcohol consumption to the risk of IC remains controversial. The purpose of this study was to assess the relation of alcohol consumption and type of beverage to the development of IC among participants in the Framingham Heart Study. Methods and Results-Alcohol consumption was categorized as 0, 1 to 6, 7 to 12, 13 to 24, and greater than or equal to 25 g/d. During a mean follow-up of 6.8 years, 414 subjects developed IC. From the lowest to the highest category of alcohol intake, the age-standardized incidence rates of IC were 5.3, 4.1, 4.2, 3.2, and 4.6 cases/1000 person-years for men and 3.4, 2.5, 1.5, 1.9, and 2.5, respectively, for women. A multivariate Cox regression model demonstrated an inverse relation, with the lowest IC risk at levels of 13 to 24 g/d for men and 7 to 12 g/d for women compared with nondrinkers;the hazard ratio (95% CI) was 0.67 (0.42 to 0.99) for men and 0.44 (0.23 to 0.80) for women. This protective effect was seen mostly with wine and beer consumption. Conclusions-Our data are consistent with a protective effect of moderate alcohol consumption on IC risk, with lowest risk observed in men consuming 13 to 24 g/d (1 to 2 drinks/d) and in women consuming 7 to 12 g/d (0.5 to 1 drink/d). C1 Boston Univ, Sch Med, Dept Med, Prevent Med & Epidemiol Sect, Boston, MA 02118 USA. NHLBI, Framingham Heart Study, Framingham, MA USA. Boston Univ, Sch Med, Gen Internal Med Sect, Boston, MA 02118 USA. Boston Univ, Sch Publ Hlth, Dept Epidemiol & Biostat, Boston, MA USA. RP Djousse, L (reprint author), Boston Univ, Sch Med, Dept Med, Prevent Med & Epidemiol Sect, 715 Albany St,Room B-612, Boston, MA 02118 USA. RI Djousse, Luc/F-5033-2017 OI Djousse, Luc/0000-0002-9902-3047 FU NHLBI NIH HHS [N01-HC-38038] NR 41 TC 34 Z9 35 U1 1 U2 4 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD DEC 19 PY 2000 VL 102 IS 25 BP 3092 EP 3097 PG 6 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 384EU UT WOS:000165930500020 PM 11120700 ER PT J AU Cooper, R Cutler, J Desvigne-Nickens, P Fortmann, SP Friedman, L Havlik, R Hogelin, G Marler, J McGovern, P Morosco, G Mosca, L Pearson, T Stamler, J Stryer, D Thom, T AF Cooper, R Cutler, J Desvigne-Nickens, P Fortmann, SP Friedman, L Havlik, R Hogelin, G Marler, J McGovern, P Morosco, G Mosca, L Pearson, T Stamler, J Stryer, D Thom, T TI Trends and disparities in coronary heart disease, stroke, and other cardiovascular diseases in the United States - Findings of the National Conference on Cardiovascular Disease Prevention SO CIRCULATION LA English DT Article DE cardiovascular diseases; epidemiology; prevention ID ACUTE MYOCARDIAL-INFARCTION; ATRIAL-FIBRILLATION; DIABETES-MELLITUS; ACE-INHIBITORS; MORTALITY; FAILURE; PATTERNS; HEALTH; PREVALENCE; PHYSICIANS AB A workshop was held September 27 through 29, 1999: to address issues relating to national trends in mortality and morbidity from cardiovascular diseases; the apparent slowing of declines in mortality from cardiovascular diseases; levels and trends in risk factors for cardiovascular diseases; disparities in cardiovascular diseases by race/ethnicity, socioeconomic status, and geography; trends in cardiovascular disease preventive and treatment services; and strategies for efforts to reduce cardiovascular diseases overall and to reduce disparities among subpopulations. The conference concluded that coronary heart disease mortality is still declining in the United States as a whole, although perhaps at a slower rate than in the 1980s; that stroke mortality rates have declined little, if at all, since 1990; and that there are striking differences in cardiovascular death rates by race/ethnicity, socioeconomic status, and geography. Trends in risk factors are consistent with a slowing of the decline in mortality; there has been little recent progress in risk factors such as smoking, physical inactivity, and hypertension control. There are increasing levels of obesity and type 2 diabetes, with major differences among subpopulations. There is considerable activity in population-wide prevention, primary prevention for higher risk people, and secondary prevention, but wide disparities exist among groups on the basis of socioeconomic status and geography, pointing to major gaps in efforts to use available, proven approaches to control cardiovascular diseases. Recommendations for strategies to attain the year 2010 health objectives were made. C1 NHLBI, NIH, Bethesda, MD 20892 USA. Loyola Univ, Med Ctr, Chicago, IL 60611 USA. Stanford Univ, Palo Alto, CA 94304 USA. NIA, Bethesda, MD 20892 USA. Ctr Dis Control & Prevent, Atlanta, GA USA. NINDS, Bethesda, MD 20892 USA. Univ Minnesota, St Paul, MN 55108 USA. Columbia Univ, New York, NY USA. Cornell Univ, New York, NY USA. Univ Rochester, Rochester, NY USA. Northwestern Univ, Sch Med, Chicago, IL USA. Agcy Healthcare Res & Qual, Rockville, MD USA. RP Friedman, L (reprint author), NHLBI, NIH, 31 Ctr Dr,MSC 2482, Bethesda, MD 20892 USA. NR 77 TC 490 Z9 499 U1 12 U2 45 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD DEC 19 PY 2000 VL 102 IS 25 BP 3137 EP 3147 PG 11 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 384EU UT WOS:000165930500027 PM 11120707 ER PT J AU Shin, HD Winkler, C Stephens, JC Bream, J Young, H Goedert, JJ O'Brien, TR Vlahov, D Buchbinder, S Giorgi, J Rinaldo, C Donfield, S Willoughby, A O'Brien, SJ Smith, MW AF Shin, HD Winkler, C Stephens, JC Bream, J Young, H Goedert, JJ O'Brien, TR Vlahov, D Buchbinder, S Giorgi, J Rinaldo, C Donfield, S Willoughby, A O'Brien, SJ Smith, MW TI Genetic restriction of HIV-1 pathogenesis to AIDS by promoter alleles of IL10 SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID IMMUNODEFICIENCY-VIRUS INFECTION; INHIBITS CYTOKINE PRODUCTION; CHEMOKINE RECEPTOR GENE; DISEASE PROGRESSION; INTERLEUKIN-10 PRODUCTION; TYPE-1 INFECTION; CHROMOSOME 6P; RESISTANCE; CCR5; LINKAGE AB IL10 is a powerful TH-2 cell cytokine produced by lymphoid cells that limits HIV-1 replication in vivo, ostensibly by inhibiting macrophage/monocyte and T-cell lymphocyte replication and secretion of inflammatory cytokines (IL1, TNF alpha, IL6, IL8, and IL12). A genetic epidemiological scan of patients enrolled in AIDS cohorts for candidate gene-linked short tandem repeat polymorphisms revealed significant genotype associations for HIV-1 infection and progression to AIDS with markers adjacent to and tracking (by linkage disequilibrium) common single nucleotide polymorphic variants in the IL10 promoter region. Individuals carrying the IL10-5'-592A (IL10-5'A) promoter allele possibly were at increased risk for HIV-1 infection, and once infected they progressed to AIDS more rapidly than homozygotes for the alternative IL10-5'-592 C/C (IL10-+/+) genotype. particularly in the later stages of HIV-1 infection. An estimated 25-30% of long-term nonprogressors (who avoid clinical AIDS for 10 or more years after HIV-1 infection) can be attributed to their IL10-+/+ promoter genotype. Alternative IL10 promoter alleles are functionally distinct in relative IL10 production, in retention of an avian erythroblastosis virus transcription factor recognition sequence and in binding to specific putative nuclear transcription factors, suggesting a potential mechanism whereby IL10-5'A down-regulation of inhibitory IL10 facilitates HIV-1 replication in vivo, accelerating the onset of AIDS. C1 NCI, Lab Genom Divers, Frederick, MD 21702 USA. NCI, Sci Applicat Int Corp, Frederick, MD 21702 USA. NCI, Expt Immunol Lab, Frederick, MD 21702 USA. NCI, Viral Epidemiol Branch, Rockville, MD 20852 USA. Johns Hopkins Sch Hyg & Publ Hlth IADS link Intra, Baltimore, MD 21205 USA. San Francisco Dept Publ Hlth, San Francisco, CA 94102 USA. Rho Inc, Chapel Hill, NC 27514 USA. Univ Calif Los Angeles, Los Angeles AIDS Inst, Sch Med, Los Angeles, CA 90095 USA. Univ Calif Los Angeles, Sch Med, Dept Med, Los Angeles, CA 90095 USA. Univ Pittsburgh, Grad Sch Publ Hlth, Pittsburgh, PA 15261 USA. NICHHD, Adolescent & Maternal AIDs Branch, NIH, Bethesda, MD 20892 USA. RP O'Brien, SJ (reprint author), NCI, Lab Genom Divers, Bldg 560,Room 21-105, Frederick, MD 21702 USA. RI Smith, Michael/B-5341-2012 NR 61 TC 211 Z9 218 U1 1 U2 4 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD DEC 19 PY 2000 VL 97 IS 26 BP 14467 EP 14472 DI 10.1073/pnas.97.26.14467 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 385FV UT WOS:000165993700087 PM 11121048 ER PT J AU Bland, ML Jamieson, CAM Akana, SF Bornstein, SR Eisenhofer, G Dallman, MF Ingraham, HA AF Bland, ML Jamieson, CAM Akana, SF Bornstein, SR Eisenhofer, G Dallman, MF Ingraham, HA TI Haploinsufficiency of steroidogenic factor-1 in mice disrupts adrenal development leading to an impaired stress response SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID NUCLEAR RECEPTOR SF-1; GLUCOCORTICOID RECEPTOR; FACTOR-I; HYPOPLASIA CONGENITA; GONADAL DEVELOPMENT; GENE; DAX-1; MUTATIONS; ENZYMES; ANXIETY AB Adrenal steroids are essential for homeostasis and survival during severe physiological stress. Analysis of a patient heterozygous for the steroidogenic factor-1 (SF-1) gene suggested that reduced expression of this nuclear receptor leads to adrenal failure. We therefore examined SF-1 heterozygous (+/-) mice as a potential model for delineating mechanisms underlying this disease. Here we show that SF-1 +/- mice exhibit adrenal insufficiency resulting from profound defects in adrenal development and organization. However, compensatory mechanisms, such as cellular hypertrophy and increased expression of the rate-limiting steroidogenic protein StAR, help to maintain adrenal function at near normal capacity under basal conditions. In contrast, adrenal deficits in SF-1 heterozygotes are revealed under stressful conditions, demonstrating that normal gene dosage of SF-1 is required for mounting an adequate stress response. Our findings predict that natural variations leading to reduced SF-1 function may underlie some forms of subclinical adrenal insufficiency, which become life threatening during traumatic stress. C1 Univ Calif San Francisco, Dept Physiol, Grad Program Biomed Sci, San Francisco, CA 94143 USA. Univ Calif San Francisco, Dept Physiol, Grad Program Dev Biol, San Francisco, CA 94143 USA. Univ Calif San Francisco, Dept Mol & Cellular Pharmacol, San Francisco, CA 94143 USA. NINDS, NIH, Bethesda, MD 20892 USA. NICHHD, NIH, Bethesda, MD 20892 USA. RP Ingraham, HA (reprint author), Univ Calif San Francisco, Dept Physiol, Grad Program Biomed Sci, 513 Parnassus,Box 0444, San Francisco, CA 94143 USA. NR 38 TC 118 Z9 118 U1 0 U2 2 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD DEC 19 PY 2000 VL 97 IS 26 BP 14488 EP 14493 DI 10.1073/pnas.97.26.14488 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 385FV UT WOS:000165993700091 PM 11121051 ER PT J AU Farrell, CM Grinberg, A Huang, SP Chen, D Pichel, JG Westphal, H Felsenfeld, G AF Farrell, CM Grinberg, A Huang, SP Chen, D Pichel, JG Westphal, H Felsenfeld, G TI A large upstream region is not necessary for gene expression or hypersensitive site formation at the mouse beta-globin locus SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID ENHANCER-BLOCKING ACTIVITY; OPEN CHROMATIN STRUCTURE; ODORANT RECEPTOR GENES; HUMAN ERYTHROID-CELLS; EMBRYONIC STEM-CELLS; TARGETED DELETION; MOLECULAR-BASIS; MICE; INSULATOR; SEQUENCE AB Developmental expression at the beta -globin locus is regulated in part by the locus control region, a region upstream of the genes containing at least five major DNase I hypersensitive sites (HSs) in mammalian erythrocytes. sequences farther 5' of these HSs are conserved in mouse and human, and both loci are embedded within a duster of functional odorant receptor genes. In humans, distant upstream sequences have been implicated in regulation of the beta -globin genes, in this study, the role of the 5'-most HSs and their adjacent sequence was investigated by deletion of an 11-kb region from the mouse locus, including 5'HS 4.2, 5'HS 5, 5'HS 6, and the 5'beta1: odorant receptor gene. Mice that were homozygous for this deletion were fully viable, and no significant effect on adult beta -globin gene expression was seen. 5'HSs 1-4, which are located downstream of the deletion, were still present in the mutant mice, in addition, two new upstream HSs, HS -60.7 and HS -62.5, were found in erythroid tissue of both wild-type and mutant mice. Therefore, although the possibility of a minor role still exists, neither the HSs nor the other regions deleted in this study are essential for beta -globin gene expression, and it is unlikely that chromatin structure is affected either upstream or downstream of the deletion. This is the largest deletion at the mouse locus control region to show no apparent phenotype, and focuses attention on the possible contribution of sequences even farther upstream. C1 NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. NICHHD, Lab Mammalian Genes & Dev, NIH, Bethesda, MD 20892 USA. RP Felsenfeld, G (reprint author), NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. NR 47 TC 30 Z9 31 U1 0 U2 0 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD DEC 19 PY 2000 VL 97 IS 26 BP 14554 EP 14559 DI 10.1073/pnas.97.26.14554 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 385FV UT WOS:000165993700102 PM 11121056 ER PT J AU Bulger, M Bender, MA van Doorninck, JH Wertman, B Farrell, CM Felsenfeld, G Groudine, M Hardison, R AF Bulger, M Bender, MA van Doorninck, JH Wertman, B Farrell, CM Felsenfeld, G Groudine, M Hardison, R TI Comparative structural and functional analysis of the olfactory receptor genes flanking the human and mouse beta-globin gene clusters SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID LOCUS-CONTROL REGIONS; OPEN CHROMATIN STRUCTURE; ODORANT RECEPTOR; ERYTHROID-CELLS; EXPRESSION; TRANSCRIPTION; ORGANIZATION; ACTIVATION; EPITHELIUM; FAMILY AB By sequencing regions flanking the beta -globin gene complex in mouse (Hbbc) and human (HBBC), we have shown that the beta -globin gene cluster is surrounded by a larger duster of olfactory receptor genes (ORGs). To facilitate sequence comparisons and to investigate the regulation of ORC expression, we have mapped 5' sequences of mRNA from olfactory epithelium encoding beta -globin-proximal ORGs. We have found that several of these genes contain multiple noncoding exons that can be alternatively spliced. Surprisingly, the only common motifs found in the promoters of these genes are a "TATA" box and a purine-rich motif. Sequence comparisons between human and mouse reveal that most of the conserved regions are confined to the coding regions and transcription units of the genes themselves, but a few blocks of conserved sequence also are found outside of ORG transcription units. The possible influence of beta -globin regulatory sequences on ORG expression in olfactory epithelium was tested in mice containing a deletion of the endogenous beta -globin locus control region, but no change in expression of the neighboring ORGs was detected. We evaluate the implications of these results for possible mechanisms of regulation of ORG transcription. C1 Fred Hutchinson Canc Res Ctr, Seattle, WA 98109 USA. Univ Washington, Sch Med, Seattle, WA 98195 USA. Columbia Univ Coll Phys & Surg, Howard Hughes Med Inst, New York, NY 10032 USA. NIDDKD, NIH, Bethesda, MD 20892 USA. Penn State Univ, Dept Biochem & Mol Biol, University Pk, PA 16802 USA. RP Groudine, M (reprint author), Fred Hutchinson Canc Res Ctr, 1100 Fairview Ave N, Seattle, WA 98109 USA. RI Hardison, Ross/G-1142-2010 OI Hardison, Ross/0000-0003-4084-7516 FU NIDDK NIH HHS [DK27635, DK44746, DK54701, R37 DK044746] NR 34 TC 53 Z9 68 U1 0 U2 0 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD DEC 19 PY 2000 VL 97 IS 26 BP 14560 EP 14565 DI 10.1073/pnas.97.26.14560 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 385FV UT WOS:000165993700103 PM 11121057 ER PT J AU Ogawa, S Chester, AE Hewitt, SC Walker, VR Gustafsson, JA Smithies, O Korach, KS Pfaff, DW AF Ogawa, S Chester, AE Hewitt, SC Walker, VR Gustafsson, JA Smithies, O Korach, KS Pfaff, DW TI Abolition of male sexual behaviors in mice lacking estrogen receptors alpha and beta (alpha beta ERKO) SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE testosterone; mounts; intromissions; ejaculation; aggression ID GENE DISRUPTION; CYP19 GENE; DEFICIENT AB Male mice with a knockout of the estrogen receptor (ER)-alpha gene, a ligand-activated transcription factor, showed reduced levels of intromissions and no ejaculations whereas simple mounting behavior was not affected, In contrast, all components of sexual behaviors were intact in male mice lacking the novel ER-beta gene. Here we measure the extent of phenotype in mice that lack both ER-alpha and ER-beta genes (alpha beta ERKO), alpha beta ERKO male mice did not show any components of sexual behaviors, including simple mounting behavior. Nor did they show ultrasonic vocalizations during behavioral tests with receptive female mice, On the other hand, reduced aggressive behaviors of alpha beta ERKO mice mimicked those of single knockout mice of ER-alpha gene (alpha ERKO), They showed reduced levels of lunge and bite aggression, but rarely showed offensive attacks. Thus, either one of the ERs is sufficient for the expression of simple mounting in male mice, indicating a redundancy in function. Offensive attacks, on the other hand, depend specifically on the ER-alpha gene. Different patterns of natural behaviors require different patterns of functions by ER genes. C1 Rockefeller Univ, Neurobiol & Behav Lab, New York, NY 10021 USA. NIEHS, Reprod & Dev Toxicol Lab, Res Triangle Pk, NC 27709 USA. Karolinska Inst, Dept Med Nutr, S-14186 Huddinge, Sweden. Univ N Carolina, Dept Pathol, Chapel Hill, NC 27599 USA. RP Ogawa, S (reprint author), Rockefeller Univ, Neurobiol & Behav Lab, 1230 York Ave,Box 275, New York, NY 10021 USA. OI Korach, Kenneth/0000-0002-7765-418X FU NICHD NIH HHS [R37 HD005751, HD-05751, R01 HD005751]; NIMH NIH HHS [MH62147-01, R01 MH062147] NR 15 TC 158 Z9 163 U1 1 U2 6 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD DEC 19 PY 2000 VL 97 IS 26 BP 14737 EP 14741 DI 10.1073/pnas.250473597 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 385FV UT WOS:000165993700133 PM 11114183 ER PT J AU Bornstein, SR Tian, H Haidan, A Bottner, A Hiroi, N Eisenhofer, G McCann, SM Chrousos, GP Roffler-Tarlov, S AF Bornstein, SR Tian, H Haidan, A Bottner, A Hiroi, N Eisenhofer, G McCann, SM Chrousos, GP Roffler-Tarlov, S TI Deletion of tyrosine hydroxylase gene reveals functional interdependence of adrenocortical and chromaffin cell system in vivo SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID RAT ADRENAL-MEDULLA; PHENYLETHANOLAMINE N-METHYLTRANSFERASE; NEUROPEPTIDE-Y; MESSENGER-RNA; TARGETED DISRUPTION; CHROMOGRANIN-A; CYCLIC-AMP; NEUROENDOCRINE REGULATION; PARAVENTRICULAR NUCLEUS; DIFFERENTIAL REGULATION AB Catecholamines are produced in the medulla of the adrenal gland and may participate in the intraglandular regulation of its cortex We analyzed the adrenal structure and function of albino tyrosine hydroxylase-null (TH-null) mice that are deficient in adrenal catecholamine production. Adrenal catecholamines were markedly reduced, and catecholamine histofluorescence was abrogated in 15-day-old TH-null mice. Chromaffin cell structure was strikingly altered at the ultrastructural level with a depletion of chromaffin vesicles and an increase in rough endoplasmic reticulum compared with wild-type mice. Remaining chromaffin vesicles lined up proximally to the cell membrane in preparation for exocytosis providing a "string-of-pearls" appearance. There was a 5-fold increase in the expression of proenkephalin mRNA(502.8 +/- 142% vs. 100 +/- 17.5%, P = 0.016) and a 2-fold increase in the expression of neuropeptide Y(213.4 +/- 41.2% vs. 100 +/- 59.9%, P = 0.014) in the TH-null animals as determined by quantitative TaqMan (Perkin-Elmer) PCR. Accordingly, immunofluorescence for met-enkephalin and neuropeptide tyrosine in these animals was strongly enhanced. The expression of phenylethanolamine IV-methyl transferase and chromogranin B mRNA was similar in TH-null and wild-type mice. In TH-null mice, adrenocortical cells were characterized by an increase in liposomes and by tubular mitochondria with reduced internal membranes, suggesting a hypofunctional state of these steroid-producing cells. In accordance with these findings, plasma corticosterone levels were decreased. plasma ACTH levels were not significantly different in TH-null mice. In conclusion, both the adrenomedullary and adrenocortical systems demonstrate structural and functional changes in catecholamine deficient TH-null mice, underscoring the great importance of the functional interdependence of these systems in vivo. C1 NICHHD, Pediat & Reprod Endocrinol Branch, NIH, Bethesda, MD 20892 USA. NINDS, NIH, Bethesda, MD 20892 USA. Univ Leipzig, Dept Pediat, D-04317 Leipzig, Germany. Louisiana State Univ, Pennington Biomed Res Ctr, Baton Rouge, LA 70808 USA. Tufts Univ, Sch Med, Dept Neurosci, Boston, MA 02111 USA. Tufts Univ, Sch Med, Dept Anat & Cell Biol, Boston, MA 02111 USA. RP Bornstein, SR (reprint author), NICHHD, Pediat & Reprod Endocrinol Branch, NIH, Bldg 10,Room 10N242,10 Ctr Dr, Bethesda, MD 20892 USA. RI Korner, Antje/B-3988-2015 OI Korner, Antje/0000-0001-6001-0356 FU NINDS NIH HHS [NS 35639] NR 57 TC 39 Z9 41 U1 0 U2 1 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD DEC 19 PY 2000 VL 97 IS 26 BP 14742 EP 14747 DI 10.1073/pnas.97.26.14742 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 385FV UT WOS:000165993700134 PM 11121073 ER PT J AU Gropman, A Levin, S Yao, L Lin, T Suchy, S Sabnis, S Hadley, D Nussbaum, R AF Gropman, A Levin, S Yao, L Lin, T Suchy, S Sabnis, S Hadley, D Nussbaum, R TI Unusual renal features of Lowe syndrome in a mildly affected boy SO AMERICAN JOURNAL OF MEDICAL GENETICS LA English DT Article DE Lowe syndrome; (oculocerebrorenal syndrome); congenital cataracts; phosphatidy linositol [PtdIns](4,5) bisphosphate; 5-phosphatase; renal tubular acidosis; glomerulonephritis; developmental delay; renal Fanconi syndrome ID OCULOCEREBRORENAL SYNDROME; MR FINDINGS; OCRL1 GENE; CARRIERS; MANIFESTATIONS; POLYMORPHISMS; 5-PHOSPHATASE; MUTATIONS AB The oculocerebrorenal syndrome of Lowe (OCRL) is an X-linked disorder characterized by congenital cataracts, mental retardation, and renal tubular dysfunction. The gene responsible for OCRL was identified by positional cloning and encodes a lipid phosphatase, phosphatidylinositol 4,5, bisphosphate [PtdIns(4,5)P2]5-phosphatase, which localizes to the Gels apparatus and is suspected to play a role in Golgi vesicular transport [Suchy et al,, 1995]. In addition to the ocular and renal manifestations, most boys with OCRL have cognitive problems and maladaptive behaviors including tantrums and stereotypies. We report a boy with a history of congenital cataracts and mild developmental delay who was also found to have hematuria with proteinuria but minimal signs of renal tubular dysfunction. Subsequent renal biopsy was compatible with a diagnosis of a noncomplement fixating chronic glomerulonephritis. Despite the atypical renal findings, skin fibroblast analysis for PtdIns (4,5)P2 5-phosphatase was performed, and enzyme activity was low, consistent with the diagnosis of OCRL. Western blot analysis from cell lysates showed the ocrl protein was decreased in size and amount. Our report shows atypical renal features of OCRL in a mildly affected boy. The possibility of OCRL should be considered in boys: with cataracts and glomerular disease, even in the absence of renal tubular defects and frank mental retardation usually associated with the syndrome, Published Wiley-Liss, Inc.'. C1 NHGRI, Med Genet Branch, NIH, Bethesda, MD 20892 USA. NICHHD, Heritable Disorders Branch, Bethesda, MD 20892 USA. Walter Reed Army Med Ctr, Dept Pediat, Washington, DC 20307 USA. NIH, Lab Genet Dis Res, Bethesda, MD 20892 USA. Armed Forces Inst Pathol, Washington, DC USA. RP Gropman, A (reprint author), NHGRI, Med Genet Branch, NIH, 10 Ctr Dr,Room 10C101, Bethesda, MD 20892 USA. NR 30 TC 15 Z9 18 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0148-7299 J9 AM J MED GENET JI Am. J. Med. Genet. PD DEC 18 PY 2000 VL 95 IS 5 BP 461 EP 466 DI 10.1002/1096-8628(20001218)95:5<461::AID-AJMG10>3.0.CO;2-D PG 6 WC Genetics & Heredity SC Genetics & Heredity GA 379GH UT WOS:000165633300010 PM 11146467 ER PT J AU Santagada, V Balboni, G Caliendo, G Guerrini, R Salvadori, S Bianchi, C Bryant, SD Lazarus, LH AF Santagada, V Balboni, G Caliendo, G Guerrini, R Salvadori, S Bianchi, C Bryant, SD Lazarus, LH TI Assessment of substitution in the second pharmacophore of Dmt-Tic analogues SO BIOORGANIC & MEDICINAL CHEMISTRY LETTERS LA English DT Article ID PHENYLETHANOLAMINE N-METHYLTRANSFERASE; DELTA-OPIOID RECEPTORS; PEPTIDE ANTAGONISTS; AROMATIC RING; ALPHA(2)-ADRENOCEPTOR; SELECTIVITY; DERIVATIVES; INHIBITORS; TOPOGRAPHY; BINDING AB The Dmt-Tic pharmacophore exhibits potent delta -opioid receptor antagonism. Analogues with substitutions in the second pharmacophore with (1, 1') or without a COOH function (2-9) were synthesized: several had high delta affinity (1', 2, 7, and 9), but exhibited low to non-selectivity toward mu receptors similar to H-Dmt-Tic-amide and H-Dmt Tic-ol. Functional bioactivity indicated high delta antagonism (pA(2) 7.4-7.9) (1', 2, and 9) and modest mu agonism, pEC(50) (6.1-6.3) (1', 2, 8, and 9), but with E-max values analogous to dermorphin. These Dmt-Tic analogues with mixed delta antagonist/mu agonist properties would appear to be better candidates as analgesics than pure mu agonists. (C) 2000 Elsevier Science Ltd. All rights reserved. C1 NIEHS, LCBRA, Res Triangle Pk, NC 27707 USA. Univ Naples, I-80134 Naples, Italy. Univ Cagliari, Dept Toxicol, I-09126 Cagliari, Italy. Univ Ferrara, Dept Pharmaceut Sci, I-44100 Ferrara, Italy. Univ Ferrara, Ctr Biotechnol, I-44100 Ferrara, Italy. Univ Ferrara, Inst Pharmacol, I-44100 Ferrara, Italy. RP Lazarus, LH (reprint author), NIEHS, LCBRA, Res Triangle Pk, NC 27707 USA. OI Guerrini, Remo/0000-0002-7619-0918; Caliendo, Giuseppe/0000-0002-5098-6045; SALVADORI, Severo/0000-0002-8224-2358 NR 20 TC 15 Z9 16 U1 0 U2 2 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0960-894X J9 BIOORG MED CHEM LETT JI Bioorg. Med. Chem. Lett. PD DEC 18 PY 2000 VL 10 IS 24 BP 2745 EP 2748 DI 10.1016/S0960-894X(00)00569-2 PG 4 WC Chemistry, Medicinal; Chemistry, Organic SC Pharmacology & Pharmacy; Chemistry GA 382WE UT WOS:000165849700013 PM 11133082 ER PT J AU Borlongan, CV AF Borlongan, CV TI Motor activity-mediated partial recovery in ischemic rats SO NEUROREPORT LA English DT Article DE cerebral infarction; enriched environment; lomocotor activity; motor asymmetry; spontaneous recovery; striatum; stroke ID MIDDLE CEREBRAL-ARTERY; ENVIRONMENTAL ENRICHMENT; NORADRENALINE DEPLETION; FUNCTIONAL RECOVERY; CELL-LINE; TRANSPLANTATION; FOREBRAIN; NEURONS; GRAFTS; DAMAGE AB Spontaneous partial recovery in motor and/or cognitive dysfunctions in stroke patients has been documented, but the factors that affect such functional improvement have not been well elucidated. The present study demonstrates that repeated behavioral testing (daily or once a week over a period of 4 weeks) promoted partial recovery from motor asymmetry in adult ischemic rats. In contrast, ischemic animals that were only tested once every 2 weeks or once after 4 weeks did not show such partial recovery. These results suggest that repeated behavioral testing (i.e., increased use of the ischemia-affected limbs and body parts) may contribute to partial recovery of motor deficits following an experimental stroke, even in the absence of pharmacological therapeutic intervention. NeuroReport 11:4063-4067 (C) 2000 Lippincott Williams & Wilkins. C1 NIDA, Dev & Plast Sect, Cellular Neurobiol Branch, NIH, Baltimore, MD 21224 USA. RP Borlongan, CV (reprint author), NIDA, Dev & Plast Sect, Cellular Neurobiol Branch, NIH, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. OI Borlongan, Cesar/0000-0002-2966-9782 NR 28 TC 22 Z9 22 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0959-4965 J9 NEUROREPORT JI Neuroreport PD DEC 18 PY 2000 VL 11 IS 18 BP 4063 EP 4067 DI 10.1097/00001756-200012180-00031 PG 5 WC Neurosciences SC Neurosciences & Neurology GA 387PU UT WOS:000166131600033 PM 11192629 ER PT J AU Blair, DG Athanasiou, M AF Blair, DG Athanasiou, M TI Ets and retroviruses - transduction and activation of members of the Ets oncogene family in viral oncogenesis SO ONCOGENE LA English DT Review DE ETS; retrovirus; oncogenesis; E26; F-MuLV; SFFV ID MURINE LEUKEMIA-VIRUS; NON-T-CELL; AVIAN ERYTHROBLASTOSIS VIRUS; CHICKEN ERYTHROID-CELLS; V-ETS; TRANSCRIPTION FACTOR; DNA-BINDING; PROVIRAL INTEGRATION; FRIEND-VIRUS; PUTATIVE ONCOGENE AB Studies of retroviral-induced oncogenesis in animal systems led to the initial discovery of viral oncogenes and their cellular homologs, and provided critical insights into their role in the neoplastic process. V-ets, the founding member of the ETS oncogene family, was originally identified as part of the fusion oncogene encoded by the avian acute leukemia virus E26 and subsequent analysis of virus induced leukemias led to the initial isolation of two other members of the ETS gene family. PU.1 was identified as a target of insertional activation in the majority of tumors induced by the murine Spleen Focus Forming virus (SFFV), while fli-1 proved to be the target of Friend murine leukemia virus (F-MuLV) in F-MuLV induced erythroleukemia, as well as that of the 10A1 and Graffi viruses. The common features of the erythroid and myeloid diseases induced by these viruses provided the initial demonstration that these and other members of the ETS family play important roles in hematopoietic development as well as disease. This review provides an overview of the role of ETS genes in retrovirally induced neoplasia, their possible mechanisms of action, and how these viral studies relate to current knowledge of the functions of these genes in hematopoiesis. C1 NCI, Frederick Canc Res & Dev Ctr, Div Basic Sci, Res Lab,Oncogene Mechanisms Sect, Frederick, MD 21702 USA. SAIC, Intramural Res Support Program, Frederick, MD 21702 USA. RP Blair, DG (reprint author), NCI, Frederick Canc Res & Dev Ctr, Div Basic Sci, Res Lab,Oncogene Mechanisms Sect, Bld 469,Room 102, Frederick, MD 21702 USA. FU NCI NIH HHS [N01-CO56000] NR 101 TC 17 Z9 17 U1 0 U2 2 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD DEC 18 PY 2000 VL 19 IS 55 BP 6472 EP 6481 DI 10.1038/sj.onc.1204046 PG 10 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 395RL UT WOS:000166595000009 PM 11175363 ER PT J AU Goldspiel, BR DeChristoforo, R Daniels, CE AF Goldspiel, BR DeChristoforo, R Daniels, CE TI A continuous-improvement approach for reducing the number of chemotherapy-related medication errors SO AMERICAN JOURNAL OF HEALTH-SYSTEM PHARMACY LA English DT Article; Proceedings Paper CT Annual Meeting of the American-Society-of-Health-System-Pharmacists CY JUN 06, 2000 CL PHILADELPHIA, PENNSYLVANIA SP Amer Soc Hlth Syst Pharmacists DE administration; antineoplastic agents; costs; devices; dosage; drug information; education; errors, medication; pharmaceutical services; pharmacy, institutional, hospital; physicians; prescribing; quality assurance; reports; toxicity ID CANCER-CHEMOTHERAPY AB A comprehensive, interdisciplinary approach for reducing the number of chemotherapy-related medication errors at the National institutes of Health Clinical Center, where approximately 8500 doses of chemotherapy agents are dispensed annually, is described. Heightened awareness of the seriousness of chemotherapy-related medication errors prompted formation of an interdisciplinary task force in June 1995 to analyze and improve the hospital's system for ordering, checking, processing, and administering cancer chemotherapy agents. Problems were analyzed and rectified in accordance with the hospital's plan-do-check-act performance-improvement model. Performance monitors for the improvements included a system to record and categorize all chemotherapy-related prescribing errors and a hospitalwide occurrence-reporting system. The task force identified seven major categories in which improvements were needed: protocol development computer-system enhancements, dose verification, information access, education for health care practitioners, error follow-up, and infusion pumps. Despite the Clinical Center's good safety-net system, 23 modifications were made to the existing system through December 1999. These changes resulted in an overall 23% decrease in prescribing errors and a 53% decrease in serious prescribing errors. The task force membership was recently broadened to include representatives of additional departments where chemotherapy agents are used, and this group recommended more than 20 additional system changes. The changes are being implemented, and their effect on reducing the number of chemotherapy-related errors will be measured. The continuous-improvement process used prospectively by the task force helps ensure that safe chemotherapy practices are instituted uniformly throughout the hospital. C1 NIH, Ctr Clin, Dept Pharm, Bethesda, MD 20892 USA. RP Goldspiel, BR (reprint author), NIH, Ctr Clin, Dept Pharm, Bldg 10,Room 1N-257,MSC 1196, Bethesda, MD 20892 USA. NR 9 TC 35 Z9 36 U1 1 U2 5 PU AMER SOC HEALTH-SYSTEM PHARMACISTS PI BETHESDA PA 7272 WISCONSIN AVE, BETHESDA, MD 20814 USA SN 1079-2082 J9 AM J HEALTH-SYST PH JI Am. J. Health-Syst. Pharm. PD DEC 15 PY 2000 VL 57 IS 24 SU 4 BP S4 EP S9 PG 6 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 385ZQ UT WOS:000166035400002 PM 11148943 ER PT J AU Kempner, ES AF Kempner, ES TI Macromolecular cross section and cellular localization: Determination by radiation target methods SO ANALYTICAL BIOCHEMISTRY LA English DT Article C1 NIAMSD, NIH, Bethesda, MD 20892 USA. RP Kempner, ES (reprint author), NIAMSD, NIH, Bethesda, MD 20892 USA. NR 15 TC 0 Z9 0 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0003-2697 J9 ANAL BIOCHEM JI Anal. Biochem. PD DEC 15 PY 2000 VL 287 IS 2 BP 191 EP 195 DI 10.1006/abio.2000.4796 PG 5 WC Biochemical Research Methods; Biochemistry & Molecular Biology; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 387YY UT WOS:000166151700001 PM 11112263 ER PT J AU Berlett, BS Levine, RL Stadtman, ER AF Berlett, BS Levine, RL Stadtman, ER TI Use of isosbestic point wavelength shifts to estimate the fraction of a precursor that is converted to a given product SO ANALYTICAL BIOCHEMISTRY LA English DT Article DE isosbestic point; spectroscopy; ferrozine; iron AB An isosbestic point is observed in overlaid spectra when a chromophoric precursor is converted to a product with a different spectrum, so that it is often assumed that an isosbestic point occurs only when the precursor is quantitatively converted to a single product. We show experimentally and by computer simulations that more complex reactions also exhibit isosbestic points and that the wavelength of the isosbestic point may change. Such wavelength changes will occur if either (i) the molar absorbtivity of the precursor changes or (ii) the fraction of the precursor that is converted to multiple products changes. In the latter case, the isosbestic wavelength and molar absorbtivities of the precursor and product can be used to calculate the fraction of the precursor that is converted to products from the relationship, f = is an element of (Precursor)(M)/is an element of (Product)(M), where f is the fractional conversion, is an element of (Precursor)(M) is the molar absorbtivity of the precursor, and is an element of (Product)(M) is the molar absorbtivity of the product. C1 NHLBI, Biochem Lab, NIH, Bethesda, MD 20892 USA. RP Stadtman, ER (reprint author), NHLBI, Biochem Lab, NIH, Bldg 3,Room 222,3 Ctr Dr,MSC-0342, Bethesda, MD 20892 USA. RI Levine, Rodney/D-9885-2011 NR 3 TC 29 Z9 31 U1 1 U2 14 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0003-2697 J9 ANAL BIOCHEM JI Anal. Biochem. PD DEC 15 PY 2000 VL 287 IS 2 BP 329 EP 333 DI 10.1006/abio.2000.4876 PG 5 WC Biochemical Research Methods; Biochemistry & Molecular Biology; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 387YY UT WOS:000166151700019 PM 11112281 ER PT J AU Radko, SP Stastna, M Chrambach, A AF Radko, SP Stastna, M Chrambach, A TI Size-dependent electrophoretic migration and separation of liposomes by capillary zone electrophoresis in electrolyte solutions of various ionic strengths SO ANALYTICAL CHEMISTRY LA English DT Article ID PARTICLES; BEHAVIOR AB The size-dependent electrophoretic migration and separation of liposomes was demonstrated and studied in capillary zone electrophoresis (CZE). The liposomes were extruded and nonextruded preparations consisting of phosphatidylcholine/phosphatidylglycerol in various ratios and ranging from 125 to 488 nm in mean diameter. When liposomes of identical surface charge density were subjected to CZE in Tris-HCI (pH 8) buffers of various ionic strengths (0.001-0.027), they migrated in order of their size. Size-dependent electrophoretic migration and separation of liposomes in CZE can be enhanced or brought about by decreasing the ionic strength of the buffer. It was shown that size-dependent migration is primarily a function of KR, where K-1 is the thickness of the electric double layer (which can be derived from the ionic strength, I, of the buffer) and R, the liposome radius. Liposome mobility depends on KR and surface charge density in a manner consistent with that expected from the Overbeek-Booth electrokinetic theory. Thus, the relaxation effect appears to be the physical mechanism underlying the size-dependent electrophoretic separation of liposomes. C1 NICHHD, Lab Cellular & Mol Biophys, Macromol Anal Sect, NIH, Bethesda, MD 20892 USA. Russian Acad Med Sci, Med Genet Res Ctr, Moscow, Russia. RP Chrambach, A (reprint author), NICHHD, Lab Cellular & Mol Biophys, Macromol Anal Sect, NIH, Bethesda, MD 20892 USA. RI Stastna, Miroslava/G-9266-2014 NR 24 TC 45 Z9 46 U1 0 U2 13 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0003-2700 J9 ANAL CHEM JI Anal. Chem. PD DEC 15 PY 2000 VL 72 IS 24 BP 5955 EP 5960 DI 10.1021/ac000661e PG 6 WC Chemistry, Analytical SC Chemistry GA 383JT UT WOS:000165879900013 PM 11140762 ER PT J AU Sinclair, PR Gorman, N Walton, HS Bement, WJ Szakacs, J Gonzalez, FJ Dalton, TP Nebert, DW Sinclair, JF AF Sinclair, PR Gorman, N Walton, HS Bement, WJ Szakacs, J Gonzalez, FJ Dalton, TP Nebert, DW Sinclair, JF TI Relative roles of CYP2E1 and CYP1A2 in mouse uroporphyria caused by acetone SO ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS LA English DT Article DE acetone; CYP1A2; CYP2E1; uroporphyrin; ethanol; uroporphyrinogen oxidation; iron ID PORPHYRIA-CUTANEA-TARDA; NULL MUTANT MICE; 5-AMINOLEVULINIC ACID; CYTOCHROME-P450 2E1; RABBIT LIVER; IRON; OXIDATION; ETHANOL; HEPATOCYTES; RAT AB Porphyria cutanea tarda is a liver disease characterized by excess production of uroporphyrin. We previously reported that acetone, an inducer of CYPSE1, enhances hepatic uroporphyrin accumulation in mice treated with iron dextran (Fe) and 5-aminolevulinic acid (ALA). Cyp3e1(-/-) mice treated with Fe and ALA were used to investigate whether CYP2E1 is required for the acetone effect. Hepatic uroporphyrin accumulation was stimulated by acetone in Cyp2e1(-/-) mice to the same extent as in wild-type mice. In the absence of acetone, uroporphyrin accumulated in Cyp2e1(-/-) mice treated with Fe and ALA, but less than in wildtype mice. However, in Cyp2a1(-/-) mice, uroporphyrin accumulation caused by Fe and ALA, with or without acetone, was completely prevented. Acetone was not an inducer of hepatic CYP1A2 in the wild-type mice. Although acetone is an inducer of CYP2E1, CYP1A2 appears to have the essential role in acetone-enhancement of uroporphyria. (C) 2000 Academic Press. C1 VA Med Ctr, White River Junction, VT 05009 USA. Dartmouth Coll, Sch Med, Dept Pharmacol Toxicol, Hanover, NH 03755 USA. Vet Adm Med Ctr, Salt Lake City, UT 84148 USA. NCI, Lab Metab, NIH, Bethesda, MD 20892 USA. Univ Cincinnati, Med Ctr, Dept Environm Hlth, Cincinnati, OH 45267 USA. Univ Cincinnati, Med Ctr, Ctr Environm Genet, Cincinnati, OH 45267 USA. RP Sinclair, PR (reprint author), VA Med Ctr, White River Junction, VT 05009 USA. FU NIEHS NIH HHS [ES 06263, ES 06231] NR 46 TC 6 Z9 6 U1 1 U2 2 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0003-9861 J9 ARCH BIOCHEM BIOPHYS JI Arch. Biochem. Biophys. PD DEC 15 PY 2000 VL 384 IS 2 BP 383 EP 390 DI 10.1006/abbi.2000.2124 PG 8 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 388AA UT WOS:000166154200022 PM 11368328 ER PT J AU Lee, CH Park, MH AF Lee, CH Park, MH TI Human deoxyhypusine synthase: interrelationship between binding of NAD and substrates SO BIOCHEMICAL JOURNAL LA English DT Article DE eIF5A; hypusine; spermidine ID LIVER ALCOHOL-DEHYDROGENASE; YEAST SACCHAROMYCES-CEREVISIAE; EUKARYOTIC CELL-PROLIFERATION; INITIATION-FACTOR 5A; HYPUSINE DEPLETION; PROTEIN SUBSTRATE; INHIBITION; ENZYME; SPERMIDINE; VIABILITY AB Deoxyhypusine synthase catalyses the NAD-dependent transfer of the butylamine moiety from the polyamine, spermidine, to a specific lysine residue of a single cellular protein, eukaryotic translation-initiation factor 5A (eIF5A) precursor. The native enzyme exists as a tetramer of four identical subunits and contains four binding sites for NAD. The binding of spermidine and NAD was studied by a filtration assay. [H-3]Spermidine binding to the enzyme was not detectable alone or in the presence of the eIF5A precursor, but was detected only in the presence of NAD or NADH, suggesting that a NAD/NADH-induced conformational change is required for the binding of spermidine. A strong NAD-dependent binding was also observed with a spermidine analogue, N-1-guanyl-1,7-diamino[H-3]heptane (GC(7)), but not with [C-14]putrescine or [C-14]spermine. Although [H-3]NAD binding to the enzyme occurred in the absence of spermidine, its affinity for the enzyme was markedly enhanced by spermidine, GC(7) and also by the eIF5A precursor. The maximum binding for NAD and spermidine was estimated to be approximate to4 molecules each/enzyme tetramer. The dependence of spermidine binding on NAD and the modulation of binding of NAD by spermidine and the eIF5A precursor suggest intricate relationships between the binding of cofactor and the substrates, and provide new insights into the reaction mechanism. C1 NIDCR, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD 20892 USA. RP Park, MH (reprint author), NIDCR, Oral & Pharyngeal Canc Branch, NIH, Bldg 30,Room 211, Bethesda, MD 20892 USA. NR 36 TC 16 Z9 17 U1 0 U2 0 PU PORTLAND PRESS PI LONDON PA 59 PORTLAND PLACE, LONDON W1N 3AJ, ENGLAND SN 0264-6021 J9 BIOCHEM J JI Biochem. J. PD DEC 15 PY 2000 VL 352 BP 851 EP 857 DI 10.1042/0264-6021:3520851 PN 3 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 388JQ UT WOS:000166177700033 PM 11104695 ER PT J AU Shen, XI Tian, ZG Holtzman, MJ Gao, B AF Shen, XI Tian, ZG Holtzman, MJ Gao, B TI Cross-talk between interleukin 1 beta (IL-1 beta) and IL-6 signalling pathways: IL-1 beta selectively inhibits IL-6-activated signal transducer and activator of transcription factor 1 (STAT1) by a proteasome-dependent mechanism SO BIOCHEMICAL JOURNAL LA English DT Article DE acute-phase protein; Janus kinase; nuclear factor kappa B; liver cells; protein tyrosine phosphatase ID NF-KAPPA-B; PROTEIN-TYROSINE-PHOSPHATASE; STIMULATED JAK/STAT PATHWAY; ADRENERGIC-RECEPTOR GENE; NEGATIVE REGULATOR; CYTOKINE RECEPTORS; GROWTH-FACTOR; KINASE; PHOSPHORYLATION; PROMOTER AB Interleukin 1 beta (IL-1 beta) suppresses the IL-6-dependent induction of type II acute-phase response genes, but the underlying mechanism for this suppression remains uncertain, Here we report that treatment of human hepatocullular carcinoma HepG2 cells with IL-1 beta inhibited the IL-6-dependent binding of signal transducer and activator of transcription factor (STAT)1, but not that of STAT3, to the high-affinity serum-inducible element ('SIE'), Furthermore, IL-1/beta, selectively down-regulated the IL-6-induced tyrosine phosphorylation of STAT1 without affecting the level of STAT1 or tyrosine phosphorylation of STAT3. Kinase assays in vitro indicated that the inhibition of STAT1 phosphorylation by IL-1 beta was not due to an upstream blockade of Janus kinase (JAK1or JAK2) activation, However, pretreatment with the proteasome inhibitor MG132 under conditions that prevented the IL-1 beta -dependent activation of the nuclear factor NF-kappaB also blocked the inhibitory effect of IL-1 beta on IL-6-activated STAT1, In related experiments, the protein tyrosine phosphatase inhibitor Na3VO4 also antagonized the inhibitory effect of IL-1 beta on the activation of STAT1 by IL-6, Taken together, these findings indicate that, by using a proteasome-dependent mechanism, IL-1 beta concomitantly induces NF-kappaB activation and dephosphorylates IL-6-activated STAT1; the latter might partly account for the inhibition by IL-1 beta of the IL-6-dependent induction of type II acute-phase genes. C1 Virginia Commonwealth Univ, Med Coll Virginia, Dept Pharmacol & Toxicol, Richmond, VA 23298 USA. Washington Univ, Sch Med, Dept Med, St Louis, MO 63110 USA. Washington Univ, Sch Med, Dept Cell Biol, St Louis, MO 63110 USA. RP Gao, B (reprint author), NIAAA, Sect Liver Biol, NIH, Flow Bldg,Room 13, Rockville, MD 20852 USA. RI Tian, Zhigang/J-3512-2013 FU NCI NIH HHS [R29CA72681]; NIAAA NIH HHS [R01AA12637, R03AA11823] NR 44 TC 26 Z9 29 U1 0 U2 0 PU PORTLAND PRESS PI LONDON PA 59 PORTLAND PLACE, LONDON W1N 3AJ, ENGLAND SN 0264-6021 J9 BIOCHEM J JI Biochem. J. PD DEC 15 PY 2000 VL 352 BP 913 EP 919 DI 10.1042/0264-6021:3520913 PN 3 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 388JQ UT WOS:000166177700041 PM 11104703 ER PT J AU Speer, AM Kimbrell, TA Wassermann, EM Repella, JD Willis, MW Herscovitch, P Post, RM AF Speer, AM Kimbrell, TA Wassermann, EM Repella, JD Willis, MW Herscovitch, P Post, RM TI Opposite effects of high and low frequency rTMS on regional brain activity in depressed patients SO BIOLOGICAL PSYCHIATRY LA English DT Article DE transcranial magnetic stimulation; regional cerebral blood flow; depressed; high frequency (20 Hz); low frequency (1 Hz); positron emission tomography ID TRANSCRANIAL MAGNETIC STIMULATION; HUMAN MOTOR CORTEX; SYNAPTIC PLASTICITY; BLOOD-FLOW; INTRAVENOUS (H2O)-O-15; PET IMAGES; EXCITABILITY; CONNECTIVITY; SUPPRESSION; HIPPOCAMPUS AB Background: High (10-20 Hz) and low frequency (1-5 Hz) repetitive transcranial magnetic stimulation (rTMS) have been explored for possible therapeutic effects in the treatment of neuropsychiatric disorders. As part of a double-blind, placebo-controlled, crossover study evaluating the antidepressant effect of daily rTMS over the left prefrontal cortex, Mle evaluated changes in absolute regional cerebral blood flow (rCBF) after treatment with 1- and 20-Hz rTMS. Based on preclinical data, we postulated that high frequency rTMS would increase and low frequency rTMS would decrease flow in frontal and related subcortical circuits, Methods: Ten medication-free, adult patients with major depression (eight unipolar and two bipolar) were serially imaged using O-15 water and Positron emission tomography to measure rCBF. Each patient was scanned at baseline and 72 hours after 10 daily treatments with 20-Hz rTMS and 10 daily treatments with 1 Hz rTMS given in a randomized order. TMS tvas administer ed over the left prefrontal cortex at 100% of motor threshold (MT). Significant changes in rCBF from pretreatment baseline tt ere determined by paired t test Results: Twenty-hertz rTMS over the left prefrontal cortex was associated only with increases in I-CBF. Significant increases in rCBF across the group of all 10 patients were located in the prefrontal cortex (L > R), the cingulate gyrus (L much greater than R), and the left amygdala, as well as bilateral insula, basal ganglia, uncus, hippocampus, parahippocampus, thalamus, and cerebellum In contrast, I-Hz rTMS was associated only with decreases in rCBF. Significant decreases in flow were noted in small areas of the right prefrontal cortex, left medial temporal cortex, left basal ganglia, and left amygdala. The changes in mood following the two rTMS frequencies were inversely related (r = -.78, p < .005, n = 10) such that individuals who improved with one frequency worsened with the other. Conclusions: These data indicate that 2 weeks of daily 20-Hz rTMS over the left prefrontal cortex at 100% MT induce persistent increases in rCBF in bilateral frontal, limbic, and paralimbic regions implicated in depression, whereas I-Hz rTMS produces more circumscribed decreases (including in the left amygdala). These data demonstrate frequency-dependent, opposite effects of high and low frequency rTMS on local and distant regional brain activity that may have important implications for clinical therapeutics in various neuropsychiatric disorders. (C) 2000 Society of Biological Psychiatry. C1 NIMH, NIH, BPB, Bethesda, MD 20895 USA. NINDS, Bethesda, MD 20892 USA. NIH, Positron Emiss Tomog, Ctr Clin, Bethesda, MD 20892 USA. Vet Adm Med Ctr, N Little Rock, AR USA. RP Post, RM (reprint author), NIMH, NIH, BPB, 10 Ctr Dr,Room 3N212, Bethesda, MD 20895 USA. NR 55 TC 277 Z9 291 U1 5 U2 22 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0006-3223 J9 BIOL PSYCHIAT JI Biol. Psychiatry PD DEC 15 PY 2000 VL 48 IS 12 BP 1133 EP 1141 DI 10.1016/S0006-3223(00)01065-9 PG 9 WC Neurosciences; Psychiatry SC Neurosciences & Neurology; Psychiatry GA 386FH UT WOS:000166049800003 PM 11137053 ER PT J AU Moldin, SO AF Moldin, SO TI Neurobiology of anxiety and fear: Challenges for genomic science of the new millennium SO BIOLOGICAL PSYCHIATRY LA English DT Editorial Material ID OBSESSIVE-COMPULSIVE DISORDER; GENE C1 NIMH, Genet Res Branch, Bethesda, MD 20892 USA. RP Moldin, SO (reprint author), NIMH, Genet Res Branch, 6001 Execut Blvd,Room 7189, Bethesda, MD 20892 USA. NR 19 TC 8 Z9 8 U1 1 U2 2 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0006-3223 J9 BIOL PSYCHIAT JI Biol. Psychiatry PD DEC 15 PY 2000 VL 48 IS 12 BP 1144 EP 1146 DI 10.1016/S0006-3223(00)00943-4 PG 3 WC Neurosciences; Psychiatry SC Neurosciences & Neurology; Psychiatry GA 386FH UT WOS:000166049800005 PM 11137055 ER PT J AU Blake, T Adya, N Kim, CH Oates, AC Zon, L Chitnis, A Weinstein, GM Liu, PP AF Blake, T Adya, N Kim, CH Oates, AC Zon, L Chitnis, A Weinstein, GM Liu, PP TI Zebrafish homolog of the leukemia gene CBFB: its expression during embryogenesis and its relationship to scl and gata-1 in hematopoiesis SO BLOOD LA English DT Article ID CORE-BINDING-FACTOR; ACUTE MYELOID-LEUKEMIA; MYOSIN HEAVY-CHAIN; TRANSCRIPTION FACTOR GATA-1; FACTOR-BETA; EMBRYONIC HEMATOPOIESIS; VERTEBRATE DEVELOPMENT; FETAL LIVER; CELL; MUTATIONS AB Mammalian CBFB encodes a transcription factor (CBF beta) that in combination with CBF alpha2 binds to specific DNA sequences and regulates expression of a number of hematopoietic genes. CBFB is associated with human leukemias through a chromosome 16 inversion and is essential for definitive hematopoiesis during mouse embryo development. We have isolated a zebrafish cbfb complementary DNA (cDNA) clone from a zebrafish kidney cDNA library. This cbfb is highly homologous to human and mouse CBFB/Cbfb genes at both the DNA and protein level. In biochemical analyses, cbf beta binds to human CBF alpha2 and enhances its DNA binding. During zebrafish development, cbfb is expressed in the lateral plate mesoderm at tail bud stage and in the intermediate cell mass (ICM, the location of embryonic hematopoiesis) between the 21- to 26-somite stages. The cbfb is also expressed in Rohon-Beard cells, cranial nerve ganglia, hindbrain, retina, branchial arches, jaw, and fin buds. Expression of cbfb is decreased or absent in the ICM and Rohon-Beard cells in some hematopoietic mutants and is unaffected in others. We have also analyzed the expression of scl and gata-1 in the same hematopoietic mutants to ascertain the relative order of these transcription factors to cbfb in zebrafish hematopoiesis. Our results indicate that cbfb is expressed in early hematopoietic progenitors and that its expression pattern in the hematopoietic mutants is similar to that of scl. (C) 2000 by The American Society of Hematology. C1 NHGRI, NIH, Bethesda, MD 20892 USA. Childrens Hosp, Howard Hughes Med Inst, Boston, MA 02115 USA. NICHHD, NIH, Bethesda, MD 20892 USA. RP Liu, PP (reprint author), NHGRI, NIH, 49 Convent Dr,Rm 3A18, Bethesda, MD 20892 USA. RI Liu, Paul/A-7976-2012 OI Liu, Paul/0000-0002-6779-025X NR 47 TC 31 Z9 32 U1 2 U2 8 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1900 M STREET. NW SUITE 200, WASHINGTON, DC 20036 USA SN 0006-4971 J9 BLOOD JI Blood PD DEC 15 PY 2000 VL 96 IS 13 BP 4178 EP 4184 PG 7 WC Hematology SC Hematology GA 380PD UT WOS:000165709500022 PM 11110689 ER PT J AU Gwynn, B Ciciotte, SL Hunter, SJ Washburn, LL Smith, RS Andersen, SG Swank, RT Dell'Angelica, EC Bonifacino, JS Eicher, EM Peters, LL AF Gwynn, B Ciciotte, SL Hunter, SJ Washburn, LL Smith, RS Andersen, SG Swank, RT Dell'Angelica, EC Bonifacino, JS Eicher, EM Peters, LL TI Defects in the cappuccino (cno) gene on mouse chromosome 5 and human 4p cause Hermansky-Pudlak syndrome by an AP-3-independent mechanism SO BLOOD LA English DT Article ID CHEDIAK-HIGASHI-SYNDROME; STORAGE POOL DEFICIENCY; BETA-GLUCURONIDASE DEFICIENCY; AP-3 ADAPTER COMPLEX; PALE EAR EP; HPS GENE; MUTATION; PROTEINS; MODELS; TRAFFICKING AB Defects in a triad of organelles (melanosomes, platelet granules, and lysosomes) result in albinism, prolonged bleeding, and lysosome abnormalities in Hermansky-Pudlak syndrome (HPS). Defects in HPS1, a protein of unknown function, and in components of the AP-3 complex cause some, but not all, cases of HPS in humans. There have been 15 inherited models of HPS described in the mouse, underscoring its marked genetic heterogeneity. Here we characterize a new spontaneous mutation in the mouse, cappuccino (cno), that maps to mouse chromosome 5 in a region conserved with human 4p15-p16. Melanosomes of cno/cno mice are immature and dramatically decreased in number in the eye and skin, resulting in severe oculocutaneous albinism. Platelet dense body contents (adenosine triphosphate, serotonin) are markedly deficient, leading to defective aggregation and prolonged bleeding. Lysosomal enzyme concentrations are significantly elevated in the kidney and liver. Genetic, immunofluorescence microscopy, and lysosomal protein trafficking studies indicate that the AP-3 complex is intact in cno/cno mice, It was concluded that the cappuccino gene encodes a product involved in an AP-3-independent mechanism critical to the biogenesis of lysosome-related organelles. (C) 2000 by The American Society of Hematology. C1 Jackson Lab, Bar Harbor, ME 04609 USA. Univ Maine, Dept Biol Sci, Orono, ME USA. Roswell Pk Canc Inst, Dept Mol & Cellular Biol, Buffalo, NY 14263 USA. NICHHD, Cell Biol & Metab Branch, NIH, Bethesda, MD 20892 USA. RP Peters, LL (reprint author), Jackson Lab, 600 Main St, Bar Harbor, ME 04609 USA. OI Bonifacino, Juan S./0000-0002-5673-6370 FU NCRR NIH HHS [RR01183]; NHLBI NIH HHS [HL31698, HL55321, R01 HL055321] NR 50 TC 30 Z9 33 U1 0 U2 0 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1900 M STREET. NW SUITE 200, WASHINGTON, DC 20036 USA SN 0006-4971 J9 BLOOD JI Blood PD DEC 15 PY 2000 VL 96 IS 13 BP 4227 EP 4235 PG 9 WC Hematology SC Hematology GA 380PD UT WOS:000165709500029 PM 11110696 ER PT J AU Mir, SS Richter, BWM Duckett, CS AF Mir, SS Richter, BWM Duckett, CS TI Differential effects of CD30 activation in anaplastic large cell lymphoma and Hodgkin disease cells SO BLOOD LA English DT Article ID NF-KAPPA-B; NECROSIS-FACTOR RECEPTOR; REED-STERNBERG CELLS; HUMAN-IMMUNODEFICIENCY-VIRUS; DOMAIN-CONTAINING RECEPTOR; DEATH-DOMAIN; T-CELLS; SIGNAL-TRANSDUCTION; CYTOPLASMIC DOMAIN; TNF RECEPTORS AB CD30 is a member of the tumor necrosis factor (TNF) receptor superfamily that is expressed on activated lymphocytes, as well as on neoplastic cells of Hodgkin disease (HD) and anaplastic large cell lymphoma (ALCL), A number of reports have shown that, depending on cellular context, CD30 signaling can exert a variety of effects, ranging from cell death to cellular proliferation, In the present study this disparity was examined, using a number of ALCL- and HD-derived cell lines. Activation of CD30 led to the induction of apoptotic death of ALCL cells, along with the selective reduction of TNF receptor-associated factor 2 and impairment in the ability of these cells to activate the pro-survival transcription factor nuclear factor kappaB (NF-kappaB), In contrast, HD cells, which constitutively express NF-kappaB, were not susceptible to CD30-induced apoptosis but could be sensitized following ectopic overexpression of a superdominant I kappaB, These studies suggest that NF-kappaB plays a determining role in the sensitivity or resistance of lymphoma cells to CD30-induced apoptosis, which may have important cons clinical treatment of CD30-positive neoplasia. (C) 2000 by The American Society of Hematology. C1 NCI, Metab Branch, Div Clin Sci, NIH, Bethesda, MD 20892 USA. RP Duckett, CS (reprint author), NCI, Metab Branch, Div Clin Sci, NIH, 10 Ctr Dr,Room 6B-05, Bethesda, MD 20892 USA. NR 57 TC 106 Z9 109 U1 0 U2 0 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1900 M STREET. NW SUITE 200, WASHINGTON, DC 20036 USA SN 0006-4971 J9 BLOOD JI Blood PD DEC 15 PY 2000 VL 96 IS 13 BP 4307 EP 4312 PG 6 WC Hematology SC Hematology GA 380PD UT WOS:000165709500039 PM 11110706 ER PT J AU Panoskaltsis-Mortari, A Taylor, PA Rubin, JS Uren, A Welniak, LA Murphy, WJ Farrel, CT Lacey, DL Blazar, BR AF Panoskaltsis-Mortari, A Taylor, PA Rubin, JS Uren, A Welniak, LA Murphy, WJ Farrel, CT Lacey, DL Blazar, BR TI Keratinocyte growth factor facilitates alloengraftment and ameliorates graft-versus-host disease in mice by a mechanism independent of repair of conditioning-induced tissue injury SO BLOOD LA English DT Article ID BONE-MARROW TRANSPLANTATION; IN-VIVO; CELLS; INTERLEUKIN-1; EXPRESSION; RECEPTOR; ENGRAFTMENT; LETHALITY; CYTOKINES; MORTALITY AB We have previously shown that pretreatment of mice with keratinocyte growth factor (KGF), an epithelial tissue repair factor, can ameliorate graft-versus-host disease (GVHD) after intensive chemoradiotherapeutic conditioning and allogeneic bone marrow transplantation (BMT). To determine whether this effect was dependent on a KGF-mediated mechanism affecting repair of conditioning-induced epithelial cell injury, we studied GVHD in the absence of conditioning using BALB/c severe combined immune-deficient (SCID) recipients given C57BL/6 T cells. KGF (5 mg/kg per day, subcutaneously) given either before or after T-cell transfer enhanced body weights and extended survival. KGF-treated recipients had elevated serum levels of the Th2 cytokine interleukin 13 (IL-13) on day 6 after T-cell transfer concomitant with reduced levels of the inflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interferon gamma (IFN-gamma). A 3-day KGF pretreatment also depressed the secondary in vitro mixed lymphocyte response (MLR) of C57BL/6 splenocytes taken 7 days after in vivo alloimmunization with irradiated BALB/c spleen cells. To determine whether KGF would inhibit host-antidonor-mediated BM rejection, pan-T-cell-depleted BALB/c BM cells were infused into sublethally irradiated C57BL/6 mice and administered KGF either before or before and after BMT. Surprisingly, all KGF schedules tested actually resulted in enhanced alloengraftment. The presence of KGF receptor on donor antihost alloreactive T cells could not be detected by binding studies with radiolabeled KGF, reverse transcriptase-polymerase chain reaction, and Western blotting. Therefore, the mechanism of action of KGF on inhibiting T-cell-mediated immune effects may not be due to a direct effect of KGF on T cells. These studies demonstrate that KGF, by mechanisms independent of repair of conditioning induced injury, has great potential as an anti-GVHD therapeutic agent with the added benefit of inhibiting the rejection of pan-T-cell-depleted donor BM allografts. (Blood. 2000;96:4350-4356) (C) 2000 by The American Society of Hematology. C1 Univ Minnesota, Dept Pediat, Div Hematol Oncol, Blood & Marrow Transplant Program, Minneapolis, MN 55455 USA. Univ Minnesota, Heme Onc BMT Div, Minneapolis, MN 55455 USA. Univ Minnesota, Ctr Canc, Minneapolis, MN 55455 USA. NCI, Bethesda, MD 20892 USA. NCI, Sci Applicat Int Corp, Frederick Canc Res & Dev Ctr, Frederick, MD 21701 USA. Amgen Inc, Thousand Oaks, CA 91320 USA. RP Panoskaltsis-Mortari, A (reprint author), Univ Minnesota, Dept Pediat, Div Hematol Oncol, Blood & Marrow Transplant Program, Box 366 Mayo,420 Delaware St SE, Minneapolis, MN 55455 USA. FU NHLBI NIH HHS [R37 HL 56067, R01 HL 63452]; NIAID NIH HHS [R01 AI 34495] NR 33 TC 71 Z9 71 U1 0 U2 4 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1900 M STREET. NW SUITE 200, WASHINGTON, DC 20036 USA SN 0006-4971 J9 BLOOD JI Blood PD DEC 15 PY 2000 VL 96 IS 13 BP 4350 EP 4356 PG 7 WC Hematology SC Hematology GA 380PD UT WOS:000165709500045 PM 11110712 ER PT J AU Mattson, MP AF Mattson, MP TI Neuroprotective signaling and the aging brain: take away my food and let me run SO BRAIN RESEARCH LA English DT Article; Proceedings Paper CT 3rd Brain Research Interactive Symposium CY NOV 02-03, 2000 CL NEW ORLEANS, LOUISIANA DE Alzheimer's disease; calories; heat shock protein; oxidative stress; mitochondria; neurotrophic factor; Parkinson's disease; stroke ID PROTECTS HIPPOCAMPAL-NEURONS; AMYLOID PRECURSOR PROTEIN; DIETARY RESTRICTION; PARKINSONS-DISEASE; DOPAMINERGIC-NEURONS; ENRICHED ENVIRONMENT; OXIDATIVE STRESS; TRANSGENIC MICE; SPATIAL MEMORY; DENTATE GYRUS AB It is remarkable that neurons are able to survive and function for a century or more in many persons that age successfully. A better understanding of the molecular signaling mechanisms that permit such cell survival and synaptic plasticity may therefore lead to the development of new preventative and therapeutic strategies for age-related neurodegenerative disorders. We all know that overrating and lack of exercise are risk factors for many different age-related diseases including cardiovascular disease, diabetes and cancers. Our recent studies have shown that dietary restriction (reduced calorie intake) can increase the resistance of neurons in the brain to dysfunction and death in experimental models of Alzheimer's disease. Parkinson's disease, Huntington's disease and stroke. The mechanism underlying the beneficial effects of dietary restriction involves stimulation of the expression of 'stress proteins' and neurotrophic factors. The neurotrophic factors induced by dietary restriction may protect neurons by inducing the production of proteins that suppress oxyradical production, stabilize cellular calcium homeostasis and inhibit apoptotic biochemical cascades. Interestingly, dietary restriction also increases numbers of newly-generated neural cells in the adult brain suggesting that this dietary manipulation can increase the brain's capacity for plasticity and self-repair. Work in other laboratories suggests that physical and intellectual activity can similarly increase neurotrophic factor production and neurogenesis. Collectively. the available data suggest the that dietary restriction, and physical and mental activity, may reduce both the incidence and severity of neurodegenerative disorders in humans. A better understanding of the cellular and molecular mechanisms underlying these effects of diet and behavior on the brain is also leading to novel therapeutic agents that mimick the beneficial effects of dietary restriction and exercise. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIA, Neurosci Lab, Gerontol Res Ctr, Baltimore, MD 21224 USA. RP Mattson, MP (reprint author), NIA, Neurosci Lab, Gerontol Res Ctr, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012 NR 51 TC 155 Z9 164 U1 4 U2 19 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0006-8993 J9 BRAIN RES JI Brain Res. PD DEC 15 PY 2000 VL 886 IS 1-2 SI SI BP 47 EP 53 DI 10.1016/S0006-8993(00)02790-6 PG 7 WC Neurosciences SC Neurosciences & Neurology GA 385WB UT WOS:000166027200005 PM 11119686 ER PT J AU Yoshidome, K Shibata, MA Couldrey, C Korach, KS Green, JE AF Yoshidome, K Shibata, MA Couldrey, C Korach, KS Green, JE TI Estrogen promotes mammary tumor development in C3(1)/SV40 large T-Antigen transgenic mice: Paradoxical loss of estrogen receptor alpha expression during tumor progression SO CANCER RESEARCH LA English DT Article ID BREAST-CANCER; MOUSE; DENSITY; GENE; HYPERPLASIA; APOPTOSIS; PROSTATE; THERAPY; GLAND; BETA AB Although several lines of epidemiological evidence exposure influences the incidence of breast cancer development, the mechanisms by which estrogen may stimulate the formation of breast cancer remain poorly understood. We have explored holy alterations in estrogen exposure can influence the development of mammary cancer in the C3(1)/T-AG transgenic model, where estrogen levels and estrogen receptor ru (ER alpha) expression do not appear to modify the level of transgene expression. The C3(1)/T-AG transgene becomes transcriptionally active in mammary ductal target cells at 3 weeks of age after the estrogen-induced differentiation of the mammary epithelial anlage to the ductal outgrowth stage. Complete maturation of the mammary ductal tree, however, is not required for cancer development because tumors arise in animals where ductal branching and terminal end bud formation have been prematurely arrested by ovariectomy. Mammary tumorigenesis in this model is promoted by increased estrogen exposure with the development of significantly more mammary intraepithelial neoplastic lesions and carcinomas associated with accelerated malignant conversion, The promotion of mammary tumors in this model appears to occur through an estrogen-induced proliferation and increase in the number of available target cells for transformation at the terminal ductal lobular units, as has been postulated to occur in women who receive hormone replacement therapy and/or by additional molecular mechanisms, Wt! show, for the first time in a transgenic mouse model, that mammary tumor progression is associated with the loss of ER alpha expression, as has been often observed in human breast cancers with important clinical significance. Estrogen signaling may, therefore, serve different functions, depending upon the stage of tumorigenesis, ERP expression is up-regulated during tumor progression, although the functional significance of this remains to be determined. C1 NCI, Lab Cell Regulat & Carcinogenesis, NIH, Inst Hlth, Bethesda, MD 20892 USA. NIEHS, Reprod & Dev Toxicol Lab, Receptor Biol Sect, Natl Inst Hlth Sci,NIH, Res Triangle Pk, NC 27709 USA. RP Green, JE (reprint author), NCI, Lab Cell Regulat & Carcinogenesis, NIH, Inst Hlth, Bldg 41,Room C629,41 Lib Dr, Bethesda, MD 20892 USA. OI Korach, Kenneth/0000-0002-7765-418X NR 28 TC 64 Z9 65 U1 1 U2 5 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD DEC 15 PY 2000 VL 60 IS 24 BP 6901 EP 6910 PG 10 WC Oncology SC Oncology GA 388UM UT WOS:000166201500028 PM 11156389 ER PT J AU Phillips, KE Herring, B Wilson, LA Rickford, MS Zhang, ML Goldman, CK Tso, JY Waldmann, TA AF Phillips, KE Herring, B Wilson, LA Rickford, MS Zhang, ML Goldman, CK Tso, JY Waldmann, TA TI IL-2R alpha-directed monoclonal antibodies provide effective therapy in a murine model of adult T-cell leukemia by a mechanism other than blockade of IL-2/IL-2R alpha interaction SO CANCER RESEARCH LA English DT Article ID I-ASSOCIATED MYELOPATHY; HUMANIZED ANTI-TAC; INTERLEUKIN-2 RECEPTOR; HTLV-I; IL-2 RECEPTOR; C RETROVIRUS; LYMPHOMA; VIRUS; TRANSPLANTATION; ACTIVATION AB Adult T-cell leukemia (ATL) develops in a small proportion of human T-cell lymphotrophic virus-1 infected individuals. The leukemia consists of an overabundance of activated T cells, which are characterized by the expression of CD25, or IL-2R alpha, on their cell surface. Presently, there is not an accepted curative therapy for ATL. We developed an in vivo model of ATL in non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice by introducing cells from an ATL patient (MET-1) into the mice. The leukemic cells proliferated in these mice that lack functional T, B, and natural killer (NK) cells. The MET-1 leukemic cells could be monitored by measurements of both serum soluble Tac (IL-2R alpha) and soluble human beta (2)-microglobulin (beta (2)mu) by ELISA. The disease progressed to death in the mice after similar to4-6 weeks. The mice developed grossly enlarged spleens and a leukemia involving ATL cells that retained the phenotype and the T-cell receptor rearrangement and human T-cell lymphotrophic virus-I integration pattern of the patient's ATL leukemia cells. This model is of value for testing the efficacy of novel therapeutic: agents for ATL. The administration of humanized anti-Tac (HAT), murine anti-Tac (MAT), and 7G7/B6, all of which target IL-2R alpha, significantly delayed the progression of the leukemia and prolonged the survival of the tumor-bearing mice. In particular, HAT induced complete remissions in 4 of 19 mice and partial remissions in the remainder. It appears that the antibodies act by a mechanism that had not been anticipated. The prevailing view is that antibodies to the IL-2R alpha receptor have their effective action by blocking the interaction of IL-2 with its growth factor receptor, thereby inducing cytokine deprivation apoptosis. However, although both HAT and MAT block the binding of IL-2 to IL3R alpha of the high affinity receptor, the 7G7/B6 monoclonal antibody binds to a different epitope on the IL-2R alpha receptor, one that is not involved in IL-2 binding. This suggested that the antibodies provide an effective therapy by a mechanism other than induction of cytokine deprivation. In accord with this view, the MET-I cells obtained from the spleens of leukemic mice did not produce IL-2, nor did they express IL-2 mRNA as assessed by reverse transcription-PCR. Another possible conventional mechanism of action involves complement-mediated killing. However, although MAT and 7G7/B6 fix rabbit complement, HAT does not do so. Furthermore, in the presence of NOD/SCID mouse serum, there was no complement-mediated lysis of MET-1 cells. In addition, the antibodies did not manifest antibody-dependent cellular cytotoxicity with NOD/SCID splenocytes that virtually lack NK cells as the effector cells as assessed in an in vitro chromium-release assay. However, in contrast to the efficacy of intact HAT, the F(ab')(2) version of this antibody was not effective in prolonging the survival of mice injected with MET-I ATL cells. In conclusion, in our murine model of ATL, monoclonal antibodies, HAT, MAT, and 7G7/B6, appear to delay progression of the leukemia by a mechanism of action that is different from the accepted mechanism of IL-2 deprivation leading to cell death. We consider two alternatives: the first, antibody-dependent cellular cytotoxicity mediated by FcRI- or FcRIII-expressing cells other than NK cells, such as monocytes or polymorphonuclear leukocytes. The second alternative we consider involves direct induction of apoptosis by the anti-IL-2R antibodies in vivo. It has been shown that the IL-2R is a critical element in the peripheral self-tolerance T-cell suicide mechanism involved in the phenomenon of activation-induced cell death. C1 NCI, Metab Branch, Div Clin Sci, NIH, Bethesda, MD 20892 USA. Prot Design Labs Inc, Fremont, CA 94555 USA. RP Waldmann, TA (reprint author), NCI, Metab Branch, Div Clin Sci, NIH, Bldg 10,Room 4N-115,10 Ctr Dr,MSC 1374, Bethesda, MD 20892 USA. NR 39 TC 65 Z9 68 U1 0 U2 1 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD DEC 15 PY 2000 VL 60 IS 24 BP 6977 EP 6984 PG 8 WC Oncology SC Oncology GA 388UM UT WOS:000166201500038 PM 11156399 ER PT J AU Sasaki, CY Lin, HC Morin, PJ Longo, DL AF Sasaki, CY Lin, HC Morin, PJ Longo, DL TI Truncation of the extracellular region abrogrates cell contact but retains the growth-suppressive activity of E-cadherin SO CANCER RESEARCH LA English DT Article ID COLON-CARCINOMA CELLS; DOMINANT-NEGATIVE CADHERIN; TRANSCRIPTION FACTOR LEF-1; MOLECULES ALPHA-CATENINS; HUMAN COLORECTAL-CANCER; BETA-CATENIN; NUCLEAR-LOCALIZATION; FACTOR RECEPTOR; GASTRIC-CANCER; GAMMA-CATENINS AB E-cadherin has been demonstrated to induce growth suppression and decrease the invasiveness of cancer cells and thus has been proposed to be a tumor suppressor gene. The ability of E-cadherin to mediate cell-cell contact and contact inhibition presumably accounts for its antitumor effects, which are attributed to the extracellular domain of the protein, Here we report that blocking the ability of E-cadherin to mediate contact inhibition by either antagonistic antibodies or expression of a mutant form of E-cadherin with the extracellular region deleted does not abrogate growth suppression. Transfection of the E-cadherin gene into the human prostate cancer cell line TSU.Pr-1 induced cell-cell contact formation, growth suppression, and redistribution of beta -catenin to the cell membrane. Treatment of the E-cadherin transfectant (CAD) with blocking antibodies disrupted cell-cell contact formation but did not influence the growth rate, suggesting that cell-cell interaction is not required for E-cadherin-mediated growth suppression, Similarly, transfection of an E-cadherin construct in which the NH2-terminal (extracellular) region was deleted did not allow cell-cell contact formation but induced growth suppression. In contrast, transfection of an E-cadherin construct in which the COOH-terminal (cytoplasmic) region was deleted did not induce suppression but promoted cell contact formation. In cells expressing E-cadherin lacking the cytoplasmic region, beta -catenin was evenly distributed in the cytoplasm, By contrast, in cells expressing E-cadherin lacking the extracellular region, beta -catenin was cell membrane associated. Growth suppression was always associated with the localization of beta -catenin to the cell membrane, The redistribution of beta -catenin from the cytoplasm to the cell membrane initially suggested the Involvement of the Wnt signaling pathway in regulating cell growth. However, only small differences in beta -catenin/T-cell factor signaling were detected in control and E-cadherin-expressing cells, suggesting that the Wnt pathway is not involved. Taken together, these findings suggest that E-cadherin-induced growth inhibition may not be solely attributed to contact inhibition but may involve the redistribution of beta -catenin from the cytoplasm to the cell membrane, and this redistribution may affect growth pathways independent of T-cell factor. C1 NIA, Immunol Lab, NIH, Baltimore, MD 21224 USA. NIA, Biol Chem Lab, NIH, Baltimore, MD 21224 USA. RP Sasaki, CY (reprint author), NIA, Immunol Lab, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 54 TC 34 Z9 35 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD DEC 15 PY 2000 VL 60 IS 24 BP 7057 EP 7065 PG 9 WC Oncology SC Oncology GA 388UM UT WOS:000166201500051 PM 11156412 ER PT J AU Wei, Q Marchler, G Edington, K Karsch-Mizrachi, I Paterson, BM AF Wei, Q Marchler, G Edington, K Karsch-Mizrachi, I Paterson, BM TI RNA interference demonstrates a role for nautilus in the myogenic conversion of Schneider cells by daughterless SO DEVELOPMENTAL BIOLOGY LA English DT Article DE RNA interference; dsRNA; myogenesis; Schneider cells; nautilus; daughterless; DMEF2 ID DOUBLE-STRANDED-RNA; SHOCK TRANSCRIPTION FACTOR; SEX DETERMINATION GENE; MUSCLE DIFFERENTIATION; MYOD FAMILY; DROSOPHILA EMBRYOGENESIS; CAENORHABDITIS-ELEGANS; DNA-BINDING; EXPRESSION; PROTEIN AB Schneider SL2 cells activate the myogenic program in response to the ectopic expression of daughterless alone, as indicated by exit from the cell cycle, syncytia formation, and the presence of muscle myosin fibrils. Myogenic conversion can be potentiated by the coexpression of DMEF2 and nautilus with daughterless. In RT-PCR assays Schneider cells express two mesodermal markers, nautilus and DMEF2 mRNAs, as well as very low levels of daughterless mRNA but no twist. Full-length RT-PCR products for nautilus and DMEF2 encode immunoprecipitable proteins. We used RNA-i to demonstrate that both endogenous nautilus expression and DMEF2 expression are required for the myogenic conversion of Schneider cells by daughterless. Coexpression of twist blocks conversion by daughterless but twist dsRNA has no effect. Our results indicate that Schneider cells are of mesodermal origin and that myogenic conversion with ectopic expression of daughterless occurs by raising the levels of daughterless protein sufficiently to allow the formation of nautilus/ daughterless heterodimers. The effectiveness of RNA-i is dependent upon protein half-life. Genes encoding proteins with relatively short half-lives (10 h), such as nautilus or HSF, are efficiently silenced, whereas more stable proteins, such as cytoplasmic actin or beta -galactosidase, are less amenable to the application of RNA-i. These results support the conclusion that nautilus is a myogenic factor in Drosophila tissue culture cells with a functional role similar to that of vertebrate MyoD. This is discussed with regard to the in vivo functions of nautilus. (C) 2000 Academic Press. C1 NCI, Biochem Lab, NIH, Bethesda, MD 20892 USA. NCI, Mol Cell Biol Lab, NIH, Bethesda, MD 20892 USA. RP Paterson, BM (reprint author), NCI, Biochem Lab, NIH, Bldg 37,Room 4A21,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 70 TC 19 Z9 20 U1 0 U2 5 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD DEC 15 PY 2000 VL 228 IS 2 BP 239 EP 255 DI 10.1006/dbio.2000.9938 PG 17 WC Developmental Biology SC Developmental Biology GA 387ZQ UT WOS:000166153300007 PM 11112327 ER PT J AU Humble, MC Kouprina, N Noskov, VN Graves, J Garner, E Tennant, RW Resnick, MA Larionov, V Cannon, RE AF Humble, MC Kouprina, N Noskov, VN Graves, J Garner, E Tennant, RW Resnick, MA Larionov, V Cannon, RE TI Radial transformation-associated recombination cloning from the mouse genome: Isolation of Tg.AC transgene with flanking DNAs SO GENOMICS LA English DT Article ID PAPILLOMA DEVELOPMENT; GLOBIN GENE; MICE; EXPRESSION; STEP; SKIN; EXTRACTION; DELETION; REAGENT; YEAST AB Transformation-associated recombination (TAR) cloning allows entire genes and large chromosomal regions to be specifically, accurately, and quickly isolated from total genomic DNA. We report the first example of radial TAR cloning from the mouse genome. Tg.AC mice carry a zeta-globin promoter/v-Ha-ras transgene. Fluorescence in situ hybridization localized the transgene integrant as a single site proximal to the centromere of chromosome 11. Radial TAR cloning in yeast was utilized to create orientation-specific yeast artificial chromosomes (YACs) to explore the possibility that cis-flanking regions were involved in transgene expression, YACs containing variable lengths of 5' or 3' banking chromosome 11 DNA and the Tg.AC transgene were specifically chosen, converted to bacterial artificial chromosomes (BACs), and assayed for their ability to promote transcription of the transgene following transfection into an FVB/N carcinoma cell line. A transgene-specific reverse transcription-polymerase chain reaction assay was utilized to examine RNA transcripts from stably transfected clones. All Tg.AC BACs expressed the transgene in this in vitro system. This report describes the cloning of the v-Ha-ras transgene and suggests that transcriptional activity may not require cis elements flanking the transgene's integration site. (C) 2000 Academic Press. C1 NIEHS, Lab Environm Carcinogenesis & Mutagenesis, Res Triangle Pk, NC 27709 USA. Univ N Carolina, Curriculum Toxicol, Chapel Hill, NC 27514 USA. NIEHS, Mol Genet Lab, Res Triangle Pk, NC 27709 USA. RP Cannon, RE (reprint author), NIEHS, Lab Environm Carcinogenesis & Mutagenesis, POB 12233,MD F1-05,111 Alexander Dr, Res Triangle Pk, NC 27709 USA. NR 27 TC 12 Z9 13 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0888-7543 J9 GENOMICS JI Genomics PD DEC 15 PY 2000 VL 70 IS 3 BP 292 EP 299 DI 10.1006/geno.2000.6384 PG 8 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA 391NL UT WOS:000166362000003 PM 11161779 ER PT J AU Pertsemlidis, A Pande, A Miller, B Schilling, P Wei, MH Lerman, MI Minna, JD Garner, HR AF Pertsemlidis, A Pande, A Miller, B Schilling, P Wei, MH Lerman, MI Minna, JD Garner, HR TI PANORAMA: An integrated web-based sequence analysis tool and its role in gene discovery SO GENOMICS LA English DT Article ID GENOMIC DNA; CPG ISLANDS; ANNOTATION; SEARCH; GENERATION; PROGRAMS AB As the exponential growth of DNA sequence information in databases continues, the task of converting this deposited information into knowledge becomes more dependent on integrative sequence analysis and visualization tools, PANORAMA is an Internet-accessible software package that performs a variety of informatics analyses on a given DNA sequence and returns a visual and interactive representation of the results. Its design is modular, so that further sequence analysis tools can be integrated with minimal effort. The utility of PANORAMA is demonstrated in the analysis of 650 kb of human genomic DNA from chromosome region 3p21.3, a region of potential tumor suppressor genes involved in lung cancer, breast cancer, and other forms of cancer. PANORAMA aided in the discovery of genes and alternate splice forms of known exons, in the demarcation of intron-exon boundaries, and in the identification of promoter regions and polymorphisms, all of which contributed to a better understanding of the region. PANORAMA is available on the World Wide Web at http://atlas.swmed.edu. (C) 2000 Academic Press. C1 Univ Texas, SW Med Ctr, Dept Biochem, Dallas, TX 75390 USA. Univ Texas, SW Med Ctr, Dept Internal Med, Dallas, TX 75390 USA. Univ Texas, SW Med Ctr, McDermott Ctr Human Growth & Dev, Dallas, TX 75390 USA. Univ Texas, SW Med Ctr, SW Grad Sch Biomed Sci, Biomed Engn Program, Dallas, TX 75390 USA. Univ Texas, SW Med Ctr, SW Med Sch, Dallas, TX 75390 USA. Univ Texas, SW Med Ctr, Genome Sci & Technol Ctr, Dallas, TX 75390 USA. Univ Texas, SW Med Ctr, Dept Pharmacol, Dallas, TX 75390 USA. Univ Texas, SW Med Ctr, Hamon Ctr Therapeut Oncol Res, Dallas, TX 75390 USA. NCI, Immunobiol Lab, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RP Garner, HR (reprint author), Univ Texas, SW Med Ctr, Dept Biochem, 5323 Harry Hines Blvd, Dallas, TX 75390 USA. OI Pertsemlidis, Alexander/0000-0003-1624-9372 FU NCI NIH HHS [CA71618, N01-CO-56000, P50 CA70907]; NHGRI NIH HHS [P50 HG01002-01] NR 23 TC 5 Z9 5 U1 0 U2 3 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0888-7543 J9 GENOMICS JI Genomics PD DEC 15 PY 2000 VL 70 IS 3 BP 300 EP 306 DI 10.1006/geno.2000.6359 PG 7 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA 391NL UT WOS:000166362000004 PM 11161780 ER PT J AU Kuan, CT Wikstrand, CJ Archer, G Beers, R Pastan, I Zalutsky, MR Bigner, DD AF Kuan, CT Wikstrand, CJ Archer, G Beers, R Pastan, I Zalutsky, MR Bigner, DD TI Increased binding affinity enhances targeting of glioma xenografts by EGFRVIII-specific scFv SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article ID SINGLE-CHAIN FV; GROWTH-FACTOR RECEPTOR; N-SUCCINIMIDYL 5-IODO-3-PYRIDINECARBOXYLATE; IMPROVED ANTIGEN-BINDING; MONOCLONAL-ANTIBODIES; RECOMBINANT IMMUNOTOXINS; TUMOR XENOGRAFTS; PHARMACOKINETICS; FRAGMENTS; VARIANT AB Combinatorial variation of CDR3 of V-H and V-L, followed by phage display, was used to select affinity mutants of the parental anti-epidermal growth factor receptor-vIII (EGFR-vIII) scFv MR1, One mutant, MR1-1(scFv), had increased specific binding affinity for EGFRvIII. It was produced and radiolabeled, and its biodistribution was evaluated in human glioma-bearing athymic mice, MR1-1 targeted the same EGFRvIII epitope as MR1 with an approximately 15-fold higher affinity (K-d 1.5 x 10(-9) M) measured by surface resonance analysis. Labeling with I-131 or I-125 was performed, and the immunoreactive fraction of the labeled MR1-1(scFv) was 50% to 55%, After incubation at 37 degreesC for 4 days, the binding affinity was maintained at 60% of initial levels. The specificity of MR1-1 for EGFRvIII was demonstrated in vitro by flow cytometry and incubation of FITC-labeled scFv with the EGFRvIII-expressing U87MG.Delta EGFR cell line or with the EGFRvIII-negative U87MG cell line in the presence or absence of competing unlabeled MR1-1(scFv), We also investigated the internalization and processing of MR1-1 compared with MR1; MR1-1 exhibited levels of both cell surface retention and internalization up to 5 times higher than those by MR1, In biodistribution studies performed in athymic mice bearing s.c. U87MG.Delta EGFR tumor xenografts, animals received paired-label intratumoral infusions of I-131-labeled MR1-1 (scFv) and I-125-labeled MR1 (scFv), Our results showed an up to 244% +/- 77% increase in tumor uptake for MR1-1 compared with that for MR1, The improved tumor retention of MR1-1(scFv) combined with its rapid clearance from normal tissues also resulted in sustained higher tumor:normal organ ratios. These results suggest that the improved affinity of MR1-1 can significantly impact in vivo glioma-specific targeting and immunotherapy, Int. J, Cancer 88:962-969, 2000, (C) 2000 Wiley-Liss, Inc. C1 Duke Univ, Med Ctr, Dept Pathol, Durham, NC 27710 USA. NCI, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. Duke Univ, Dept Radiol, Med Ctr, Durham, NC USA. RP Bigner, DD (reprint author), Duke Univ, Med Ctr, Dept Pathol, Durham, NC 27710 USA. FU NCI NIH HHS [CA11898, CA42324]; NINDS NIH HHS [NS20023] NR 35 TC 53 Z9 54 U1 0 U2 4 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD DEC 15 PY 2000 VL 88 IS 6 BP 962 EP 969 DI 10.1002/1097-0215(20001215)88:6<962::AID-IJC20>3.0.CO;2-U PG 8 WC Oncology SC Oncology GA 379AT UT WOS:000165619300020 PM 11093822 ER PT J AU Engels, EA Sinclair, MD Biggar, RJ Whitby, D Ebbesen, P Goedert, JJ Gastwirth, JL AF Engels, EA Sinclair, MD Biggar, RJ Whitby, D Ebbesen, P Goedert, JJ Gastwirth, JL TI Latent class analysis of human herpesvirus 8 assay performance and infection prevalence in sub-Saharan Africa and Malta SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article ID SARCOMA-ASSOCIATED HERPESVIRUS; KAPOSIS-SARCOMA; DIAGNOSTIC-TESTS; PERIPHERAL-BLOOD; DNA-SEQUENCES; AIDS EPIDEMIC; ERROR RATES; HUMAN-HERPESVIRUS-8; TRANSMISSION; ANTIBODIES AB Human herpesvirus 8 (HHV-8) is thought to be highly prevalent in Mediterranean countries and sub-Saharan Africa, where it causes Kaposi's sarcoma in a small proportion of infected immunocompetent persons. However, the lack of serological tests with established accuracy has hindered our understanding of the prevalence, risk factors and natural history of HHV-8 infection. We tested 837 subjects from Congo, Botswana (mostly young adults) and Malta (elderly adults), using an immunofluorescence assay and 2 enzyme immunoassays (EIAs, to viral proteins K8.1 and orf65). Each assay found HHV-8 seroprevalence to be high (49-87%) in the African populations and generally lower (9-54%) in Malta. However, there was only modest agreement among tests regarding which subjects were seropositive (3-way kappa, 0.05-0.34). We used latent class analysis to model this lack of agreement, estimating each test's sensitivity and specificity and each population's HHV-8 prevalence. Using this approach, the K8.1 EIA had consistently high sensitivity (91-100%) and specificity (92-100%) across populations, suggesting that it might be useful for epidemiological studies. Compared with the K8.1 EIA, both the immunofluorescence assay and the orf65 EIA Rad more variable sensitivity (80-100% and 58-87%, respectively) and more variable specificity (57-100% and 48-85%, respectively). HHV-8 prevalence was 7% among elderly Maltese adults. Prevalence was much higher (82%) in Congo, consistent with very high Kaposi's sarcoma incidence there. Prevalence was also high in Botswana (87% in Sans, an indigenous group, and 76% in Bantus), though Kaposi's sarcoma is not common, suggesting that additional co-factors besides HHV-8 are needed for development of Kaposi's sarcoma. Int. J. Cancer 88:1003-1008, 2000, Published 2000 Wiley-Liss, Inc.dagger C1 NCI, Div Canc Epidemol & Genet, Viral Epidemiol Branch, Rockville, MD 20852 USA. Math Policy Res Inc, Princeton, NJ USA. Sci Applicat Int Corp, Frederick, MD USA. George Washington Univ, Dept Stat, Washington, DC 20052 USA. NCI, Div Canc Epidemiol & Genet, Biostat Branch, Rockville, MD USA. Danish Canc Soc, Dept Virus & Canc, DK-8000 Aarhus, Denmark. RP Engels, EA (reprint author), NCI, Div Canc Epidemol & Genet, Viral Epidemiol Branch, 6120 Execut Blvd,EPS 8005, Rockville, MD 20852 USA. NR 47 TC 50 Z9 55 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD DEC 15 PY 2000 VL 88 IS 6 BP 1003 EP 1008 DI 10.1002/1097-0215(20001215)88:6<1003::AID-IJC26>3.0.CO;2-9 PG 6 WC Oncology SC Oncology GA 379AT UT WOS:000165619300026 PM 11093828 ER PT J AU Stover, E Steinberg, L AF Stover, E Steinberg, L TI Early detection of HIV: Implications for prevention research SO JOURNAL OF ACQUIRED IMMUNE DEFICIENCY SYNDROMES LA English DT Editorial Material C1 NIMH, Div Mental Disorders Behav Res & AIDS, NIH, Bethesda, MD 20892 USA. RP Stover, E (reprint author), NIMH, Div Mental Disorders Behav Res & AIDS, NIH, 6001 Execut Blvd, Bethesda, MD 20892 USA. NR 0 TC 2 Z9 2 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1525-4135 J9 J ACQ IMMUN DEF SYND JI J. Acquir. Immune Defic. Syndr. PD DEC 15 PY 2000 VL 25 SU 2 BP S93 EP S93 DI 10.1097/00126334-200012152-00001 PG 1 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 386TN UT WOS:000166077800001 ER PT J AU Lin, FLM Majumdar, A Klotz, LC Reszka, AP Neidle, S Seidman, MM AF Lin, FLM Majumdar, A Klotz, LC Reszka, AP Neidle, S Seidman, MM TI Stability of DNA triplexes on shuttle vector plasmids in the replication pool in mammalian cells SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HELIX-FORMING OLIGONUCLEOTIDES; GUANINE-RICH OLIGONUCLEOTIDES; POTASSIUM-MEDIATED INHIBITION; MURINE C-PIM-1 PROTOONCOGENE; POTENTIAL ANTICANCER AGENTS; KI-RAS PROMOTER; 3RD STRAND; DUPLEX DNA; TRANSCRIPTION ELONGATION; INTRACELLULAR STABILITY AB Triple helix-forming oligonucleotides may be useful as gene-targeting reagents in vivo, for applications such as gene knockout. One important property of these complexes is their often remarkable stability, as demonstrated in solution and in cells following transfection, Although encouraging, these measurements do not necessarily report tripler stability in cellular compartments that support DNA functions such as replication and mutagenesis. We have devised a shuttle vector plasmid assay that reports the stability of triplexes on DNA that undergoes replication and mutagenesis. The assay is based on plasmids with novel variant supF tRNA genes containing embedded sequences for tripler formation and psoralen cross-linking. Triple helix-forming oligonucleotides were linked to psoralen and used to form triplexes on the plasmids, At various times after introduction into cells, the psoralen was activated by exposure to long wave ultraviolet light (WA). After time for replication and mutagenesis, progeny plasmids were recovered and the frequency of plasmids with mutations in the supF gene determined. Site-specific mutagenesis by psoralen cross-links was dependent on precise placement of the psoralen by the triple helix-forming oligonucleotide at the time of WA treatment. The results indicated that both pyrimidine and purine motif triplexes were much less stable on replicated DNA than on DNA in vitro or in total transfected DNA. Incubation of cells with amidoanthraquinone-based tripler stabilizing compounds enhanced the stability of the pyrimidine triplex. C1 NIA, Mol Genet Lab, NIH, Baltimore, MD 21224 USA. LCK Associates, Gloucester, MA 01930 USA. Inst Canc Res, Chester Beatty Labs, London SW3 6JB, England. RP Seidman, MM (reprint author), NIA, Mol Genet Lab, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 74 TC 13 Z9 13 U1 1 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 15 PY 2000 VL 275 IS 50 BP 39117 EP 39124 DI 10.1074/jbc.M005404200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 384PB UT WOS:000165953100025 PM 10993885 ER PT J AU Wang, XT Martindale, JL Holbrook, NJ AF Wang, XT Martindale, JL Holbrook, NJ TI Requirement for ERK activation in cisplatin-induced apoptosis SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID EPIDERMAL GROWTH-FACTOR; SIGNAL-REGULATED KINASE; PROGRAMMED CELL-DEATH; PROTEIN-KINASE; CYTOCHROME-C; OXIDATIVE STRESS; CANCER CELLS; INDUCED CYTOTOXICITY; MAMMALIAN-CELLS; FACTOR RECEPTOR AB Cisplatin activates multiple signal transduction pathways involved in coordinating cellular responses to stress, Here we demonstrate a requirement for extracellular signal-regulated protein kinase (ERK), a member of the mitogen-activated protein kinase family in mediating cisplatin-induced apoptosis of human cervical carcinoma HeLa cells. Cisplatin treatment resulted in dose- and time- dependent activation of ERK, That elevated ERK activity contributed to cell death by cisplatin was supported by several observations: 1) PD98059 and U0126, chemical inhibitors of the MEK/ERK signaling pathway, prevented apoptosis; 2) pretreatment of cells with TPA, an activator of the ERK pathway, enhanced their sensitivity to cisplatin; 3) suramin, a growth factor receptor antagonist that greatly suppressed ERK activation, likewise inhibited cisplatin-induced apoptosis; and, finally, 4) HeLa cell variants selected for cisplatin resistance showed reduced activation of ERK following cisplatin treatment. Cisplatin-induced apoptosis was associated with cytochrome c release and subsequent caspase-3 activation, both of which could be prevented by treatment with the MEK inhibitors. However, the caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone protected HeLa cells against apoptosis without affecting ERK activation. Taken together, our findings suggest that ERK activation plays an active role in mediating cisplatin-induced apoptosis of HeLa cells and functions upstream of caspase activation to initiate the apoptotic signal. C1 NIA, Cell Stress & Aging Sect, Biol Chem Lab, NIH, Baltimore, MD 21224 USA. RP Holbrook, NJ (reprint author), NIA, Cell Stress & Aging Sect, Biol Chem Lab, NIH, Box 12,5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 54 TC 473 Z9 482 U1 2 U2 23 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 15 PY 2000 VL 275 IS 50 BP 39435 EP 39443 DI 10.1074/jbc.M004583200 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 384PB UT WOS:000165953100068 PM 10993883 ER PT J AU Ohashi, E Bebenek, K Matsuda, T Feaver, WJ Gerlach, VL Friedberg, EC Ohmori, H Kunkel, TA AF Ohashi, E Bebenek, K Matsuda, T Feaver, WJ Gerlach, VL Friedberg, EC Ohmori, H Kunkel, TA TI Fidelity and processivity of DNA synthesis by DNA polymerase kappa, the product of the human DINB1 gene SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HIV-1 REVERSE-TRANSCRIPTASE; NUCLEOTIDE INCORPORATION; XERODERMA-PIGMENTOSUM; ESCHERICHIA-COLI; BASE SUBSTITUTION; MOUSE HOMOLOGS; MINOR-GROOVE; MUTAGENESIS; BETA; REPLICATION AB Mammalian DNA polymerase kappa (pol kappa), a member of the UmuC/DinB nucleotidyl transferase superfamily, has been implicated in spontaneous mutagenesis. Here we show that human pol kappa copies undamaged DNA with average single-base substitution and deletion error rates of 7 x 10(-3) and 2 x 10(-3), respectively. These error rates are high when compared to those of most other DNA polymerases. pol kappa also has unusual error specificity, producing a high proportion of T.CMP mispairs and deleting and adding non-reiterated nucleotides at extraordinary rates. Unlike other members of the UmuC/DinB family, pol kappa can processively synthesize chains of 25 or more nucleotides. This moderate processivity may reflect a contribution of C-terminal residues, which include two zinc clusters. The very low fidelity and moderate processivity of pol kappa is novel in comparison to any previously studied DNA polymerase, and is consistent with a role in spontaneous mutagenesis. C1 NIEHS, Struct Biol Lab, Res Triangle Pk, NC 27709 USA. NIEHS, Mol Genet Lab, Res Triangle Pk, NC 27709 USA. Kyoto Univ, Inst Virus Res, Sakyo Ku, Kyoto 6068507, Japan. Univ Texas, SW Med Ctr, Dept Pathol, Lab Mol Pathol, Dallas, TX 75390 USA. RP Kunkel, TA (reprint author), NIEHS, Struct Biol Lab, POB 12233,111 TW Alexander Dr, Res Triangle Pk, NC 27709 USA. FU NCI NIH HHS [CA-69029, CA-75733] NR 45 TC 176 Z9 180 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 15 PY 2000 VL 275 IS 50 BP 39678 EP 39684 DI 10.1074/jbc.M005309200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 384PB UT WOS:000165953100096 PM 11006276 ER PT J AU Elizondo, G Corchero, J Sterneck, E Gonzalez, FJ AF Elizondo, G Corchero, J Sterneck, E Gonzalez, FJ TI Feedback inhibition of the retinaldehyde dehydrogenase gene ALDH1 by retinoic acid through retinoic acid receptor A and CCAAT/enhancer-binding protein beta SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ALDEHYDE DEHYDROGENASE; C/EBP-BETA; RESPONSE ELEMENT; TRANSCRIPTIONAL REGULATION; MAMMALIAN-CELLS; PROMOTER; IDENTIFICATION; ALPHA; EXTRACTION; ACTIVATION AB Aldehyde dehydrogenase 1 (ALDH1) plays a major role in the biosynthesis of retinoic acid (RA), a hormone required for several essential life processes. Recent evidence, using the aryl hydrocarbon receptor-null mouse, suggests that elevated hepatic RA down-regulates ALDH1 in a unique feedback pathway to control RA biosynthesis. To determine the mechanism of suppression of the ALDH1 gene by RA, transactivation studies were carried out in Hepa-1 mouse hepatoma cells. RA decreased expression of an ALDH1-CAT construct containing -2536 base pairs of DNA upstream of the transcription start site. Retinoic acid receptor alpha (RAR alpha) transactivates the ALDH1 gene promoter through a complex with an RA response-like element (RARE) located at -91/-75 bp, which bound to the RAR alpha /retinoid X receptor beta heterodimer. CCAAT/enhancer-binding protein (C/EBP beta) also transactivates the ALDH1 gene promoter through a CCAAT box located 3' and directly adjacent to the RARE, and the ALDH1 gene is downregulated in C/EBP beta -null mouse liver. Exposure of Hepa-1 cells to RA results in a decrease in C/EBP beta mRNA levels; however, there was no difference in mRNA and protein levels between wild-type and AHR-null mouse liver. These data support a model in which the RAR alpha and C/EBP beta activate the ALDH1 gene promoter through the RARE and C/EBP response elements, and in Hepa-1 cells, high levels of RA inhibit this activation by decreasing cellular levels of C/EBP beta. C1 NCI, NIH, Lab Metab, Bethesda, MD 20892 USA. NCI, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RP Gonzalez, FJ (reprint author), NCI, NIH, Lab Metab, Bldg 37,Rm 3E-24, Bethesda, MD 20892 USA. NR 35 TC 49 Z9 49 U1 0 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 15 PY 2000 VL 275 IS 50 BP 39747 EP 39753 DI 10.1074/jbc.M004987200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 384PB UT WOS:000165953100105 PM 10995752 ER PT J AU Ogilvie, M Yu, XB Nicolas-Metral, V Pulido, SM Liu, C Ruegg, UT Noguchi, CT AF Ogilvie, M Yu, XB Nicolas-Metral, V Pulido, SM Liu, C Ruegg, UT Noguchi, CT TI Erythropoietin stimulates proliferation and interferes with differentiation of myoblasts SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID DYSTROPHIC MOUSE MUSCLE; HELIX-LOOP-HELIX; STEM-CELL-FACTOR; ENDOTHELIAL-CELLS; SIGNAL-TRANSDUCTION; ERYTHROID-CELLS; CYTOKINE RECEPTORS; PROLACTIN RECEPTOR; CALCIUM CHANNELS; SKELETAL-MUSCLE AB Erythropoietin (Epo) is required for the production of mature red blood cells. The requirement for Epo and its receptor (EpoR) for normal heart development and the response of vascular endothelium and cells of neural origin to Epo provide evidence that the function of Epo as a growth factor or cytokine to protect cells from apoptosis extends beyond the hematopoietic lineage. me now report that the EpoR is expressed on myoblasts and can mediate a biological response of these cells to treatment with Epo. Primary murine satellite cells and myoblast C2C12 cells, both of which express endogenous EpoR, exhibit a proliferative response to Epo and a marked decrease in terminal differentiation to form myotubes. me also observed that Epo stimulation activates Jak2/Stat5 signal transduction and increases cytoplasmic calcium, which is dependent on tyrosine phosphorylation. In erythroid progenitor cells, Epo stimulates induction of transcription factor GATA-1 and EpoR; in C2C12 cells, GATA-3 and EpoR expression are induced. The decrease in differentiation of C2C12 cells is concomitant with an increase in Myf-5 and MyoD expression and inhibition of myogenin induction during differentiation, altering the pattern of expression of the MyoD family of transcription factors during muscle differentiation. These data suggest that, rather than acting in an instructive or specific mode for differentiation, Epo can stimulate proliferation of myoblasts to expand the progenitor population during differentiation and may have a potential role in muscle development or repair. C1 NIDDK, Biol Chem Lab, NIH, Bethesda, MD 20892 USA. Univ Lausanne, Sch Pharm, Pharmacol Grp, CH-1015 Lausanne, Switzerland. RP Noguchi, CT (reprint author), NIDDK, Biol Chem Lab, NIH, Bethesda, MD 20892 USA. OI RUEGG, Urs/0000-0001-6078-8280 NR 60 TC 172 Z9 180 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 15 PY 2000 VL 275 IS 50 BP 39754 EP 39761 DI 10.1074/jbc.M004999200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 384PB UT WOS:000165953100106 PM 10995753 ER PT J AU Oka, H Harada, K Suzuki, M Ito, Y AF Oka, H Harada, K Suzuki, M Ito, Y TI Separation of spiramycin components using high-speed counter-current chromatography SO JOURNAL OF CHROMATOGRAPHY A LA English DT Article DE counter-current chromatography; spiramycins; macrolide antibiotics; antibiotics AB High-speed counter-current chromatography was successfully applied to the separation of the main components of spiramycin. A 25-mg quantity of sample was separated using a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-water (3:6:5:5), and the fractions were analyzed by high-performance liquid chromatography and fast atom bombardment mass spectrometry. The separation yielded 13.4, 0.7 and 1.7 mg of spiramycins I, II, and III with purities of 98.2, 92.3 and 97.4%, respectively. (C) 2000 Elsevier Science BN. All rights reserved. C1 Aichi Prefectural Inst Publ Hlth, Kita Ku, Nagoya, Aichi 4628576, Japan. Meijo Univ, Fac Pharm, Tempaku Ku, Nagoya, Aichi 4688503, Japan. NHLBI, Biophys Chem Lab, NIH, Bethesda, MD 20892 USA. RP Oka, H (reprint author), Aichi Prefectural Inst Publ Hlth, Kita Ku, Nagoya, Aichi 4628576, Japan. NR 11 TC 30 Z9 35 U1 3 U2 5 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0021-9673 J9 J CHROMATOGR A JI J. Chromatogr. A PD DEC 15 PY 2000 VL 903 IS 1-2 BP 93 EP 98 DI 10.1016/S0021-9673(00)00903-1 PG 6 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 384ZU UT WOS:000165977800010 PM 11153959 ER PT J AU Fitzhugh, DJ Naik, S Caughman, SW Hwang, ST AF Fitzhugh, DJ Naik, S Caughman, SW Hwang, ST TI Cutting edge: C-C chemokine receptor 6 is essential for arrest of a subset of memory T cells on activated dermal microvascular endothelial cells under physiologic flow conditions in vitro SO JOURNAL OF IMMUNOLOGY LA English DT Article ID ADHESION MOLECULE; E-SELECTIN; LYMPHOCYTES; SKIN; IDENTIFICATION; RECOGNITION; EXPRESSION; CULTURE; ANTIGEN; CLONING AB Memory T cells (mTC) express multiple chemokine receptors (including CCR4 and CCR6) that may potentially be involved in their arrest on inflamed endothelia, Herein, we specifically addressed whether CCR6 Is required for mTC to arrest on TNF-a-activated human dermal microvascular endothelial cells (HDMEC) in vitro under shear stress conditions, Recombinant liver and activation-regulated chemokine (LARC)/CCL20 (a CCR6 ligand) induced firm arrest of cutaneous lymphocyte Ag+ mTC in a flow chamber system using purified substrates, Strikingly, desensitization of CCR6 with LARC, but not thymus and activation-regulated chemokine/CCL17 or secondary lymphoid tissue chemokine/CCL21, caused a 50-75% decrease (p < 0.001) in arrest of mTC on HDMEC, which was indistinguishable from the reduction observed when total mTC were treated with pertussis toxin (p > 0.5), CCR6-depleted mTC also had a markedly reduced ability to arrest on HDMEC. Our results suggest that LARC production by activated endothelial cells and CCR6 expression by mTC may be critical components In the pertussis toxin-sensitive arrest of mTC on activated HDMEC. C1 NCI, Dermatol Branch, Bethesda, MD 20892 USA. Emory Univ, Sch Med, Dept Dermatol, Atlanta, GA 30332 USA. RP Hwang, ST (reprint author), NCI, Dermatol Branch, Bldg 10,Room 12N246,10 Ctr Dr MSC 1908, Bethesda, MD 20892 USA. NR 28 TC 89 Z9 91 U1 1 U2 2 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD DEC 15 PY 2000 VL 165 IS 12 BP 6677 EP 6681 PG 5 WC Immunology SC Immunology GA 381XZ UT WOS:000165790500001 PM 11120783 ER PT J AU Apolloni, E Bronte, V Mazzoni, A Serafini, P Cabrelle, A Segal, DM Young, HA Zanovello, P AF Apolloni, E Bronte, V Mazzoni, A Serafini, P Cabrelle, A Segal, DM Young, HA Zanovello, P TI Immortalized myeloid suppressor cells trigger apoptosis in antigen-activated T lymphocytes SO JOURNAL OF IMMUNOLOGY LA English DT Article ID TUMOR-INDUCED IMMUNOSUPPRESSION; COLONY-STIMULATING FACTOR; NITRIC-OXIDE; BONE-MARROW; IN-VITRO; MURINE MACROPHAGES; DENDRITIC CELLS; MICE; INTERLEUKIN-4; PROLIFERATION AB We described a generalized suppression of CTL anamnestic responses that occurred in mice bearing large tumor nodules or immunized with powerful recombinant viral immunogens. Immune suppression entirely depended on GM-CSF-driven accumulation of CD11b(+)/Gr-1(+) myeloid suppressor cells (MSC) in secondary lymphoid organs. To further investigate the nature and properties of MSG, we immortalized CD11b(+)/Gr-1(+) tells isolated from the spleens of immunosuppressed mice, using a retrovirus encoding the v-myc and v-mf oncogenes. Immortalized cells expressed monocyte/macrophage markers (CD11b, F4/80, CD86, CD11c), but they differed from previously characterized macrophage lines in their capacities to inhibit T lymphocyte activation. Two MSC lines, MSC-1 and MSC-2, were selected based upon their abilities to inhibit Ag-specific proliferative and functional CTL responses. MSC-1 line was constitutively inhibitory, while suppressive functions of MSC-2 line were stimulated by exposure to the cytokine IL-4, Both MSC lines triggered the apoptotic cascade in Ag-activated T lymphocytes by a mechanism requiring cell-cell contact. Some well-known membrane molecules involved in the activation of apoptotic pathways (e.g., TNF-related apoptosis-inducing ligand, Fas ligand, TNF-alpha) were ruled out as candidate effecters for the suppression mechanism. The immortalized myeloid lines represent a novel, useful tool to shed light on the molecules involved in the differentiation of myeloid-related suppressors as well as in the inhibitory pathway they use to control T lymphocyte activation. C1 Univ Padua, Dept Oncol & Surg Sci, Oncol Sect, I-35128 Padua, Italy. NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. NCI, Frederick Canc Res & Dev Ctr, Expt Immunol Lab, Frederick, MD 21702 USA. RP Bronte, V (reprint author), Univ Padua, Dept Oncol & Surg Sci, Oncol Sect, Via Gattamelata 64, I-35128 Padua, Italy. RI Serafini, Paolo/C-8195-2012; Bronte, Vincenzo/K-7902-2016 OI Serafini, Paolo/0000-0002-3651-9176; Bronte, Vincenzo/0000-0002-3741-5141 NR 33 TC 104 Z9 108 U1 0 U2 4 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD DEC 15 PY 2000 VL 165 IS 12 BP 6723 EP 6730 PG 8 WC Immunology SC Immunology GA 381XZ UT WOS:000165790500008 PM 11120790 ER PT J AU Chung, DH Natarajan, K Boyd, LF Tormo, J Mariuzza, RA Yokoyama, WM Margulies, DH AF Chung, DH Natarajan, K Boyd, LF Tormo, J Mariuzza, RA Yokoyama, WM Margulies, DH TI Mapping the ligand of the NK inhibitory receptor Ly49A on living cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID MHC CLASS-I; NATURAL-KILLER-CELLS; CRYSTAL-STRUCTURE; PEANUT AGGLUTININ; HLA-E; CD94/NKG2 RECEPTORS; PEPTIDE DEPENDENCY; LY-49A RECOGNIZES; DIRECT BINDING; COMPLEX AB We have used a recombinant, biotinylated form of the mouse NK cell inhibitory receptor, Ly49A, to visualize the expression of MHC class I (MHC-I) ligands on living lymphoid cells. A panel of murine strains, including MHC congenic lines, was examined. We detected binding of Ly49A to cells expressing H-2D(d), H-2D(k), and H-2D(P) but not to those expressing other MHC molecules, Cells of the MHC-recombinant strain B10,PL (M-2(u)) not only bound Ly49A but also inhibited cytolysis by Ly49A(+) effector cells, consistent with the correlation of in vitro binding and NK cell function. Binding of Ly49A to H-2D(d)-bearing cells of different lymphoid tissues was proportional to the level of H-2Dd expression and was not related to the lineage of the cells examined, These binding results, interpreted in the context of amino acid sequence comparisons and the recently determined three-dimensional structure of the Ly49A/H-2D(d) complex, suggest a role for amino acid residues at the amino-terminal end of the alpha1 helix of the MHC-I molecule for Ly49A interaction. This view is supported by a marked decrease in affinity of an H-2D(d) mutant, 152 M, for Ly49A. Thus, allelic variation of MHC-I molecules controls measurable affinity for the NK inhibitory receptor Ly49A and explains differences in functional recognition in different mouse strains. C1 NIAID, Immunol Lab, Mol Biol Sect, NIH, Bethesda, MD 20892 USA. CSIC, Ctr Nacl Biotecnol, Madrid, Spain. Univ Maryland, Ctr Adv Res Biotechnol, Rockville, MD 20850 USA. Washington Univ, Sch Med, Howard Hughes Med Inst, Div Rheumatol, St Louis, MO 63110 USA. RP Margulies, DH (reprint author), NIAID, Immunol Lab, Mol Biol Sect, NIH, Bldg 10,Room 11N311, Bethesda, MD 20892 USA. RI Chung, Doo Hyun/J-2791-2012; Margulies, David/H-7089-2013; OI Margulies, David/0000-0001-8530-7375 NR 72 TC 28 Z9 28 U1 0 U2 2 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD DEC 15 PY 2000 VL 165 IS 12 BP 6922 EP 6932 PG 11 WC Immunology SC Immunology GA 381XZ UT WOS:000165790500036 PM 11120818 ER PT J AU Wyllie, DH Kiss-Toth, E Visintin, A Smith, SC Boussouf, S Segal, DM Duff, GW Dower, SK AF Wyllie, DH Kiss-Toth, E Visintin, A Smith, SC Boussouf, S Segal, DM Duff, GW Dower, SK TI Evidence for an accessory protein function for toll-like receptor 1 in anti-bacterial responses SO JOURNAL OF IMMUNOLOGY LA English DT Article ID CELL WALL COMPONENTS; CUTTING EDGE; NEISSERIA-MENINGITIDIS; LIPOPOLYSACCHARIDE RESPONSIVENESS; INNATE IMMUNITY; RECOGNITION; ACTIVATION; GENE; INTERLEUKIN-1; ENDOTOXIN AB Members of the Toll-like receptor (TLR) family are components of the mammalian anti-microbial response, signaling with a domain closely related to that of IL-L receptors, In this report the expression and function of TLR1, a TLR of unknown function, are examined. TLR1 is expressed by monocytes, as demonstrated using a novel mAb. Monocytes also express TLR2, TLR1 transfection of HeLa cells, which express neither TLR1 nor TLR2, was not sufficient to confer responsiveness to several microbial extracts. However, cotransfection of TLR1 and TLR2 resulted in enhanced signaling by HeLa cells to soluble factors released from Neisseria meningitidis relative to the response with either TLR alone. This phenomenon was also seen with high concentrations of some preparations of LPS, The N, meningitidis factors recognized by TLR1/TLR2 were not released by N, meningitidis mutant in the LpxA gene. Although LpxA is required for LPS biosynthesis, because cooperation between TLR1 and TLR2 was not seen with all LPS preparations, the microbial component(s) TLR1/2 recognizes is likely to be a complex of LPS and other molecules or a compound metabolically and chemically related to LPS, The functional IL-1R consists of a heterodimer; this report suggests a similar mechanism for TLR1 and TLR2, for certain agonists, These data further suggest that mammalian responsiveness to some bacterial products may be mediated by combinations of TLRs, suggesting a mechanism for diversifying the repertoire of Toll-mediated responses. C1 Univ Sheffield, Royal Hallamshire Hosp, Funct Genom Grp, Div Mol & Genet Med, Sheffield S10 2JF, S Yorkshire, England. NCI, NIH, Expt Immunol Branch, Bethesda, MD 20892 USA. RP Dower, SK (reprint author), Univ Sheffield, Royal Hallamshire Hosp, Funct Genom Grp, Div Mol & Genet Med, M Floor, Sheffield S10 2JF, S Yorkshire, England. RI Kiss-Toth, Endre/A-8596-2014 OI Kiss-Toth, Endre/0000-0003-4406-4017 NR 52 TC 196 Z9 205 U1 4 U2 6 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD DEC 15 PY 2000 VL 165 IS 12 BP 7125 EP 7132 PG 8 WC Immunology SC Immunology GA 381XZ UT WOS:000165790500061 PM 11120843 ER PT J AU Onda, M Willingham, M Wang, QC Kreitman, RJ Tsutsumi, Y Nagata, S Pastan, I AF Onda, M Willingham, M Wang, QC Kreitman, RJ Tsutsumi, Y Nagata, S Pastan, I TI Inhibition of TNF-alpha produced by Kupffer cells protects against the nonspecific liver toxicity of immunotoxin anti-Tac(Fv)-PE38, LMB-2 SO JOURNAL OF IMMUNOLOGY LA English DT Article ID TUMOR-NECROSIS-FACTOR; AERUGINOSA EXOTOXIN-A; PSEUDOMONAS EXOTOXIN; RECOMBINANT IMMUNOTOXINS; COMPLETE REGRESSION; DNA FRAGMENTATION; DIPHTHERIA-TOXIN; MICE; MACROPHAGES; EXPRESSION AB LMB-2 (anti-Tac(Fv)-PE3B) is a recombinant immunotoxin composed of the Fv fragment of the anti-Tac Ab fused to a 38-kDa form of Pseudomonas exotoxin A. Recent clinical trials showed that LMB-2 is a promising agent for the treatment of patients with Tac-positive leukemia or lymphoma. One major side effect that needs to be overcome is nonspecific liver toxicity. In the current study, we have analyzed the mechanism of this toxicity using a mouse model. Mice that were injected with a lethal dose of LMB-2 showed severe hepatic necrosis, Immunohistochemistry revealed that LMB-2 accumulated in Kupffer cells in the liver, suggesting that the damage to the hepatocytes was indirect, When we examined the effects of LMB-2, on peritoneal macrophages, cells in the same lineage as Kupffer cells, we found that LMB-2 induced the production of TNF-alpha by these cells. Following LMB-2 administration to mice, the levels of TNF-alpha in the liver increased to very high levels, whereas the rise in serum levels was modest. in addition, the LMB-2-induced liver toxicity was blocked by a specific TNF binding protein (TNFsRp55). Liver toxicity was also blocked by indomethacin, which also blocked the rise of TNF-alpha in the liver. Both TNFsRp55 and indomethacin treatment protected mice against a lethal dose of LMB-2, These data indicate that TNF-alpha produced in the liver by Kupffer cells has an important causal role in the nonspecific liver toxicity of LMB-2, These findings have important clinical implications for the use of immunotoxins in the therapy of patients with cancer. C1 NCI, Mol Biol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. Wake Forest Univ, Sch Med, Dept Pathol, Winston Salem, NC 27106 USA. RP Pastan, I (reprint author), NCI, Mol Biol Lab, Div Basic Sci, NIH, Bldg 37,Room 4E17,37 Convent Dr,MSC 4255, Bethesda, MD 20892 USA. NR 26 TC 51 Z9 52 U1 0 U2 0 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD DEC 15 PY 2000 VL 165 IS 12 BP 7150 EP 7156 PG 7 WC Immunology SC Immunology GA 381XZ UT WOS:000165790500064 PM 11120846 ER PT J AU Hinton, DM Vuthoori, S AF Hinton, DM Vuthoori, S TI Efficient inhibition of Escherichia coli RNA polymerase by the bacteriophage T4 AsiA protein requires that AsiA binds first to free sigma(70) SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE AsiA; sigma(70); RNA polymerase; inhibition; transcription ID TRANSCRIPTIONAL ACTIVATOR MOTA; MIDDLE PROMOTER; SIGMA-70 SUBUNIT; DNA; MUTANT; GENE; POLYPEPTIDES; SPECIFICITY; RECOGNITION; REGIONS AB The bacteriophage T4 AsiA protein inhibits transcription from host and phage early promoters and is required, along with the T4 MotA protein, for activation of phage middle promoters. During infection, AsiA is found in a tight association with the sigma (70) subunit of RNA polymerase. We show that AsiA binds rapidly to free sigma (70) at either 4 degreesC or 30 degreesC to form an AsiA-sigma (70) complex that with core efficiently reconstitutes the AsiA-inhibited RNA polymerase. In contrast, AsiA does not inhibit transcription after a 15 minute incubation with RNA polymerase holoenzyme at 4 degreesC, and at 30 degreesC an incubation of several minutes is required to inhibit most of the polymerase. We show that the heat step needed for AsiA is not the formation of an active AsiA protein. However, it is consistent with the momentary dissociation of holoenzyme to give free sigma (70) and core. Our results indicate that AsiA is either unable to access holoenzyme directly or does so very slowly. Efficient generation of the AsiA-inhibited RNA polymerase requires that AsiA first binds to free sigma (70) and then the AsiA-sigma (70) complex binds to core to form the AsiA-inhibited polymerase. C1 NIDDKD, Mol & Cellular Biol Lab, NIH, Bethesda, MD 20892 USA. RP Hinton, DM (reprint author), NIDDKD, Mol & Cellular Biol Lab, NIH, Bldg 8,Room 2A-13, Bethesda, MD 20892 USA. NR 31 TC 28 Z9 28 U1 0 U2 0 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-2836 J9 J MOL BIOL JI J. Mol. Biol. PD DEC 15 PY 2000 VL 304 IS 5 BP 731 EP 739 DI 10.1006/jmbi.2000.4113 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 385AZ UT WOS:000165980600006 PM 11188759 ER PT J AU Belyantseva, IA Frolenkov, GI Wade, JB Mammano, F Kachar, B AF Belyantseva, IA Frolenkov, GI Wade, JB Mammano, F Kachar, B TI Water permeability of cochlear outer hair cells: Characterization and relationship to electromotility SO JOURNAL OF NEUROSCIENCE LA English DT Article DE mechanosensory transduction; electromotility; water permeability; aquaporins; voltage-dependent capacitance; postnatal development; organ of Corti; patch clamp ID ELECTROKINETIC SHAPE CHANGES; MAJOR INTRINSIC PROTEIN; EXTERNAL ELECTRIC-FIELD; LENS FIBER MEMBRANES; GUINEA-PIG COCHLEA; POSTNATAL-DEVELOPMENT; VOLUME REGULATION; INNER-EAR; RED-CELLS; RAT AB The distinguishing feature of the mammalian outer hair cells (OHCs) is to elongate and shorten at acoustic frequencies, when their intracellular potential is changed. This "electromotility" or "electromechanics" depends critically on positive intracellular pressure (turgor), maintained by the inflow of water through yet uncharacterized water pathways. We measured the water volume flow, J(v), across the plasma membrane of isolated guinea pig and rat OHCs after osmotic challenges and estimated the osmotic water permeability coefficient, P-f, to be similar to 10(-2) cm/sec. This value is within the range reported for osmotic flow mediated by the water channel proteins, aquaporins. J(v) was inhibited by HgCl2, which is known to block aquaporin-mediated water transport. P-f values that were estimated for OHCs from neonatal rats were of the order of similar to 2x10(-3) cm/sec, equivalent to that of membranes lacking water channel proteins. In an immunofluorescence assay we showed that an anti-peptide antibody specific for aquaporins labels the lateral plasma membrane of the OHC in the region in which electromotility is generated. Using patch-clamp recording, we found that water influx into the OHC is regulated by intracellular voltage. We also found that the most pronounced increases of the electromotility-associated charge movement and of the expression of OHC water channels occur between postnatal days 8 and 12, preceding the onset of hearing function in the rat. Our data indicate that electromotility and water transport in OHCs may influence each other structurally and functionally. C1 Natl Inst Deafness & Other Commun Disorders, Sect Struct Cell Biol, NIH, Bethesda, MD 20892 USA. Univ Maryland, Sch Med, Dept Physiol, Baltimore, MD 21201 USA. Int Sch Adv Studies, Ist Nazl Fis Mat, I-34014 Trieste, Italy. Int Sch Adv Studies, Biophys Lab, I-34014 Trieste, Italy. RP Kachar, B (reprint author), Natl Inst Deafness & Other Commun Disorders, Sect Struct Cell Biol, NIH, Bldg 36,Room 5D15, Bethesda, MD 20892 USA. RI Mammano, Fabio/I-5064-2012 OI Mammano, Fabio/0000-0003-3751-1691 NR 57 TC 34 Z9 35 U1 0 U2 6 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD DEC 15 PY 2000 VL 20 IS 24 BP 8996 EP 9003 PG 8 WC Neurosciences SC Neurosciences & Neurology GA 384ZE UT WOS:000165976500008 PM 11124975 ER PT J AU Valverde, O Maldonado, R Valjent, E Zimmer, AM Zimmer, A AF Valverde, O Maldonado, R Valjent, E Zimmer, AM Zimmer, A TI Cannabinoid withdrawal syndrome is reduced in pre-proenkephalin knock-out mice SO JOURNAL OF NEUROSCIENCE LA English DT Article DE cannabinoid; opioid; mice; mutation; withdrawal; addiction; tolerance ID OPIATE WITHDRAWAL; CB1; MORPHINE; RECEPTORS; OPIOIDS; BRAIN; PAIN; DELTA-9-TETRAHYDROCANNABINOL; PHARMACOLOGY; TRANSMISSION AB The functional interactions between the endogenous cannabinoid and opioid systems were evaluated in pre-proenkephalin-deficient mice. Antinociception induced in the tail-immersion test by acute Delta9-tetrahydrocannabinol was reduced in mutant mice, whereas no difference between genotypes was observed in the effects induced on body temperature, locomotion, or ring catalepsy. During a chronic treatment with Delta9-tetrahydrocannabinol, the development of tolerance to the analgesic responses induced by this compound was slower in mice lacking enkephalin. In addition, cannabinoid withdrawal syndrome, precipitated in Delta9-tetrahydrocannabinol-dependent mice by the injection of SR141716A, was significantly attenuated in mutant mice. These results indicate that the endogenous enkephalinergic system is involved in the antinociceptive responses of Delta9-tetrahydrocannabinol and participates in the expression of cannabinoid abstinence. C1 NIMH, Genet Lab, Bethesda, MD 20892 USA. Univ Pompeu Fabra, Fac Ciencies Salut & Vida, Neuropharmacol Lab, Barcelona 08003, Spain. RP Maldonado, R (reprint author), Univ Bonn, Psychiat Clin, Dept Mol Neurobiol, Sigmund Freud Str 25, D-53105 Bonn, Germany. RI Valverde, Olga/D-8654-2012; Zimmer, Andreas/B-8357-2009; Maldonado, Rafael/F-5657-2014 OI Maldonado, Rafael/0000-0002-4359-8773 NR 42 TC 72 Z9 76 U1 1 U2 1 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD DEC 15 PY 2000 VL 20 IS 24 BP 9284 EP 9289 PG 6 WC Neurosciences SC Neurosciences & Neurology GA 384ZE UT WOS:000165976500040 PM 11125007 ER PT J AU LeBeau, AP Van Goor, F Stojilkovic, SS Sherman, A AF LeBeau, AP Van Goor, F Stojilkovic, SS Sherman, A TI Modeling of membrane excitability in gonadotropin-releasing hormone-secreting hypothalamic neurons regulated by Ca2+ mobilizing and adenylyl cyclase-coupled receptors SO JOURNAL OF NEUROSCIENCE LA English DT Article DE GT1 neurons; mathematical modeling; voltage-gated calcium entry; calcium-mobilization; phospholipase C; adenylyl cyclase ID ACTIVATED CALCIUM CURRENT; OPERATED HTRP3 CHANNELS; PANCREATIC BETA-CELLS; CURRENT I-CRAC; ENDOPLASMIC-RETICULUM; STORE DEPLETION; GNRH RELEASE; OSCILLATIONS; INACTIVATION; MITOCHONDRIA AB Gonadotropin-releasing hormone (GnRH) secretion from native and immortalized hypothalamic neurons is regulated by endogenous Ca2+-mobilizing and adenylyl cyclase (AC)-coupled receptors. Activation of both receptor types leads to an increase in action potential firing frequency and a rise in the intracellular Ca2+ concentration ([Ca2+](i)) and neuropeptide secretion. The stimulatory action of Ca2+-mobilizing agonists on voltage-gated Ca2+ influx is determined by depletion of the intracellular Ca2+ pool, whereas AC agonist-stimulated Ca2+ influx occurs independently of stored Ca2+ and is controlled by cAMP, possibly through cyclic nucleotide-gated channels. Here, experimental records from immortalized GnRH-secreting neurons are simulated with a mathematical model to determine the requirements for generating complex membrane potential (V-m) and [Ca2+](i) responses to Ca2+-mobilizing and AC agonists. Included in the model are three pacemaker currents: a store-operated Ca2+ current (I-SOC), an SK-type Ca2+-activated K+ current (I-SK), and an inward current that is modulated by cAMP and [Ca2+](i) (I-d). Spontaneous electrical activity and Ca2+ signaling in the model are predominantly controlled by I-d, which is activated by cAMP and inhibited by high [Ca2+](i). Depletion of the intracellular Ca2+ pool mimics the receptor-induced activation of I-SOC and I-SK, leading to an increase in the firing frequency and Ca2+ influx after a transient cessation of electrical activity. However, increasing the activity of I-d simulates the experimental response to forskolin- induced activation of AC. Analysis of the behaviors of I-SOC, I-d, and I-SK in the model reveals the complexity in the interplay of these currents that is necessary to fully account for the experimental results. C1 NIDDKD, Math Res Branch, NIH, Bethesda, MD 20892 USA. NICHHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. RP LeBeau, AP (reprint author), NIDDKD, Math Res Branch, NIH, BSA Bldg,Suite 350,9190 Rockville Pike MSC 2690, Bethesda, MD 20892 USA. NR 44 TC 38 Z9 38 U1 0 U2 0 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD DEC 15 PY 2000 VL 20 IS 24 BP 9290 EP 9297 PG 8 WC Neurosciences SC Neurosciences & Neurology GA 384ZE UT WOS:000165976500041 PM 11125008 ER PT J AU Thompson, AC Zapata, A Justice, JB Vaughan, RA Sharpe, LG Shippenberg, TS AF Thompson, AC Zapata, A Justice, JB Vaughan, RA Sharpe, LG Shippenberg, TS TI kappa-Opioid receptor activation modifies dopamine uptake in the nucleus accumbens and opposes the effects of cocaine SO JOURNAL OF NEUROSCIENCE LA English DT Article DE kappa-opioid receptors; dopamine; dopamine uptake; cocaine; nucleus accumbens; striatum; quantitative microdialysis; rotating disk electrode voltammetry; autoradiography; Western blot; rats ID PROTEIN-KINASE-C; BEHAVIORAL SENSITIZATION; EXTRACELLULAR DOPAMINE; QUANTITATIVE MICRODIALYSIS; LIGAND-BINDING; EXTRACTION FRACTION; NOR-BINALTORPHIMINE; PHORBOL ESTERS; TIME COURSE; TRANSPORTER AB Coadministration of kappa -opioid receptor agonists (kappa -agonists) with cocaine prevents alterations in dialysate dopamine (DA) concentration in the nucleus accumbens (Acb) that occur during abstinence from repeated cocaine treatment. Quantitative microdialysis was used to determine the mechanism producing these effects. Rats were injected with cocaine (20 mg/kg, i.p.), or saline, and the selective kappa -agonist U-69593 (0.32 mg/kg, s.c.), or vehicle, once daily for 5 d. Extracellular DA concentration (DA(ext)) and extraction fraction (E-d), an indirect measure of DA uptake, were determined 3 d later. Repeated cocaine treatment increased E-d, whereas repeated U-69593 treatment decreased E-d, relative to controls. Coadministration of both drugs yielded intermediate Ed values not different from controls. In vitro DA uptake assays confirmed that repeated U-69593 treatment produces a dose-related, region-specific decrease in DA uptake and showed that acute U-69593 administration increases DA uptake in a norbinaltorphimine reversible manner. Repeated U-69593 also led to a decrease in [I-125] RTI-55 binding to the DA transporter (DAT), but did not decrease total DAT protein. These results demonstrate that kappa -opioid receptor activation modulates DA uptake in the Acb in a manner opposite to that of cocaine: repeated U-69593 administration decreases the basal rate of DA uptake, and acute U-69593 administration transiently increases DA uptake. kappa -agonist treatment also alters DAT function. The action of kappa -agonists on DA uptake or DAT binding, or both, may be the mechanism(s) mediating the previously reported "cocaine-antagonist" effect of kappa -opioid receptor agonists. C1 NIDA, Behav Neurosci Branch, NIH, Baltimore, MD 21224 USA. Emory Univ, Dept Chem, Atlanta, GA 30322 USA. Univ N Dakota, Sch Med & Hlth Sci, Dept Biochem & Mol Biol, Grand Forks, ND 58202 USA. RP Thompson, AC (reprint author), SUNY Buffalo, Dept Psychol, Behav Neurosci Program, Pk Hall, Buffalo, NY 14260 USA. FU NIDA NIH HHS [DA10896, DA11176] NR 58 TC 89 Z9 90 U1 1 U2 5 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD DEC 15 PY 2000 VL 20 IS 24 BP 9333 EP 9340 PG 8 WC Neurosciences SC Neurosciences & Neurology GA 384ZE UT WOS:000165976500046 PM 11125013 ER PT J AU Belyantseva, IA Adler, HJ Curi, R Frolenkov, GI Kachar, B AF Belyantseva, IA Adler, HJ Curi, R Frolenkov, GI Kachar, B TI Expression and localization of prestin and the sugar transporter GLUT-5 during development of electromotility in cochlear outer hair cells SO JOURNAL OF NEUROSCIENCE LA English DT Article DE mechanosensory transduction; unconventional cell motility; motor protein; organ of Corti; postnatal development; voltage-dependent capacitance ID ELECTROKINETIC SHAPE CHANGES; MOTILITY; MOTOR; INHIBITION; AMPLIFIER; MECHANISM; INNER AB Electromotility, i.e., the ability of cochlear outer hair cells (OHCs) to contract and elongate at acoustic frequencies, is presumed to depend on the voltage-driven conformational changes of "motor" proteins present in the OHC lateral plasma membrane. Recently, two membrane proteins have been proposed as candidates for the OHC motor. A sugar transporter, GLUT-5, was proposed based on its localization in the OHCs and on the observation that sugar transport alters the voltage sensitivity of the OHC motor mechanism. Another candidate, "prestin," was identified from a subtracted OHC cDNA library and shown to impart voltage-driven shape changes to transfected cultured cells. We used antibodies specific for these two proteins to show that they are highly expressed in the lateral membrane of OHCs. We also compared the postnatal expression patterns of these proteins with the development of electromotility in OHCs of the apical turn of the rat organ of Corti. The patch-clamp recording of transient charge movement associated with electromotility indicates that half of the maximal expression of the motor protein occurs at postnatal day 9. Prestin incorporation in the plasma membrane begins from postnatal day 0 and increases progressively in a time course coinciding with that of electromotility. GLUT-5 is not incorporated into the lateral plasma membrane of apical OHCs until postnatal day 15. Our results suggest that, although GLUT-5 may be involved in the control of electromotility, prestin is likely to be a fundamental component of the OHC membrane motor mechanism. C1 Natl Inst Deafness & Other Commun Disorders, Sect Struct Cell Biol, NIH, Bethesda, MD 20892 USA. Univ Sao Paulo, Inst Biomed Sci, Dept Physiol & Biophys, BR-05508 Sao Paulo, Brazil. RP Kachar, B (reprint author), Natl Inst Deafness & Other Commun Disorders, Sect Struct Cell Biol, NIH, Bldg 36,Room 5D15, Bethesda, MD 20892 USA. FU NIDCD NIH HHS [Z01 DC 0002-11] NR 29 TC 134 Z9 139 U1 1 U2 8 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD DEC 15 PY 2000 VL 20 IS 24 BP art. no. EP RC116 PG 5 WC Neurosciences SC Neurosciences & Neurology GA 384ZE UT WOS:000165976500002 PM 11125015 ER PT J AU Weiss, MD Hammer, J Quarles, RH AF Weiss, MD Hammer, J Quarles, RH TI Oligodendrocytes in aging mice lacking myelin-associated glycoprotein are dystrophic but not apoptotic SO JOURNAL OF NEUROSCIENCE RESEARCH LA English DT Article DE 2 ',3 '-cyclic nucleotide 3 '-phosphodiesterase; (CNPase); myelin; MAG-null mice; neural cell adhesion molecule (NCAM); oligodendrogliopathy ID MULTIPLE-SCLEROSIS LESIONS; CELL-ADHESION MOLECULE; CENTRAL-NERVOUS-SYSTEM; FYN TYROSINE KINASE; ANTERIOR MEDULLARY VELUM; BASIC-PROTEIN; MONOCLONAL-ANTIBODY; N-CAM; RAT; DEFICIENT AB Although MAG-null mice myelinate relatively normally except for subtle structural abnormalities in the periaxonal region of myelin sheaths, they develop more severe pathological changes as they age. The purpose of this study was to further define the biochemical aspects of CNS pathology caused by an absence of MAG. Proteins associated with myelin and oligodendrocytes were quantified by densitometry of western blots in whole brain homogenates, as well as in isolated myelinated axons and myelin. Neither myelin yields, nor levels of myelin basic protein and proteolipid protein, were decreased in comparison to control levels in 14-month-old MAG-null mice. On the other hand, 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNPase) and the 120 kD neural cell adhesion molecule (N-CAM) were substantially reduced in whole brain, myelinated axons, and myelin. Tubulin, Na(+)K(+)ATPase and Fyn tyrosine kinase were also reduced significantly in myelin-related fractions, but not in whole brain homogenate. The decreased levels of these proteins suggest pathological abnormalities in oligodendrocytes. Furthermore, significant reductions of CNPase and 120 kD NCAM were also present at 2 months, indicating that the oligodendroglial abnormalities begin at a relatively early age. Neither TUNEL assays nov multiplex RT-PCR for mRNAs of apoptosis-related proteins in the aging MAG-null mice provided evidence for apoptotic oligodendrocytes. These biochemical findings suggest oligodendroglial damage in MAG-null mice and support the morphological observations pointing to a progressive "dying-back oligodendrogliopathy" as a consequence of MAG deficiency. (C) 2000 Wiley-Liss, Inc. C1 NINDS, Mol & Cellular Neurobiol Lab, NIH, Myelin & Brain Dev Sect, Bethesda, MD 20892 USA. RP Weiss, MD (reprint author), NINDS, Mol & Cellular Neurobiol Lab, NIH, Myelin & Brain Dev Sect, Bldg 49,Room 2A28, Bethesda, MD 20892 USA. NR 52 TC 22 Z9 22 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0360-4012 J9 J NEUROSCI RES JI J. Neurosci. Res. PD DEC 15 PY 2000 VL 62 IS 6 BP 772 EP 780 DI 10.1002/1097-4547(20001215)62:6<772::AID-JNR3>3.0.CO;2-X PG 9 WC Neurosciences SC Neurosciences & Neurology GA 379ND UT WOS:000165648200003 PM 11107161 ER PT J AU Chen, Z Mocharla, VP Farmer, JM Pettit, GR Hamel, E Pinney, KG AF Chen, Z Mocharla, VP Farmer, JM Pettit, GR Hamel, E Pinney, KG TI Preparation of new anti-tubulin ligands through a dual-mode, addition-elimination reaction to a bromo-substituted alpha,beta-unsaturated sulfoxide SO JOURNAL OF ORGANIC CHEMISTRY LA English DT Article ID ANTIMITOTIC NATURAL-PRODUCTS; COMBRETASTATIN A-4; ANTINEOPLASTIC AGENTS; TUMOR VASCULATURE; DIARYL COMPOUNDS; COLCHICINE SITE; INHIBITION; PHOSPHATE; POLYMERIZATION; BINDING C1 Baylor Univ, Dept Chem & Biochem, Waco, TX 76798 USA. Arizona State Univ, Canc Res Inst, Tempe, AZ 85287 USA. Arizona State Univ, Dept Chem, Tempe, AZ 85287 USA. NCI, Frederick Canc Res & Dev Ctr, Screening Technol Branch, Dev Therapeut Program,Div Canc Treatment & Diag, Frederick, MD 21702 USA. RP Pinney, KG (reprint author), Baylor Univ, Dept Chem & Biochem, POB 97348, Waco, TX 76798 USA. RI Farmer, Matt/C-2571-2016; OI Farmer, Matt/0000-0002-7279-1847; Pinney, Kevin/0000-0003-1262-8631 FU NCI NIH HHS [CA44344-06-11] NR 29 TC 31 Z9 31 U1 0 U2 0 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0022-3263 J9 J ORG CHEM JI J. Org. Chem. PD DEC 15 PY 2000 VL 65 IS 25 BP 8811 EP 8815 DI 10.1021/jo0004761 PG 5 WC Chemistry, Organic SC Chemistry GA 381FQ UT WOS:000165752000054 PM 11112609 ER PT J AU Wang, DS Armstrong, DL AF Wang, DS Armstrong, DL TI Tetraethylammonium potentiates the activity of muscarinic potassium channels in guinea-pig atrial myocytes SO JOURNAL OF PHYSIOLOGY-LONDON LA English DT Article ID K-ATP CHANNELS; BETA-GAMMA-SUBUNITS; CELL-FREE MEMBRANE; ACETYLCHOLINE ACTIVATION; ION CHANNELS; G-PROTEINS; MECHANISM; RECEPTORS; NA+; DETERMINANTS AB 1. The modulation of native muscarinic potassium channels (K-ACh) by tetraethylammonium (TEA) was studied at. 35 degreesC in cell-free patches from acutely dissociated guinea-pig atrial myocytes. The channels were identified unambiguously by their conductance, inward rectification, rapid gating kinetics and pharmacological responses to muscarinic agonists and GTP gammaS. 2. Addition of 5 mM TEA to the cytoplasmic side of the patches almost doubled the open probability of KACh channels that had been activated maximally by GTP gammaS. In contrast even 30 mM TEA did not significantly potentiate the response to carbachol in whole-cell recordings. 3. Unlike GTP gammaS, TEA alone did not activate K-ACh channels de novo, but in patches that showed spontaneous KACh activity, 5 mM TEA increased channel open probability fourfold in the absence of added sodium, ATP or guanine nucleotides. Furthermore, the effect of TEA was not blocked by 10 muM atropine or by 1 mM GDP betaS, and subsequent addition of 0.1 mM GTP gammaS did not stimulate channel activity further in the presence of TEA. 4. Phosphatidylinositol 4,5-bisphosphate (PIP2) also stimulates K-ACh channels under these conditions, but the kinetics of gating differ from channels stimulated by either TEA or GTP, which are very similar to one another. 5. The effects of TEA were not mimicked by tetramethyl- or tetrapentylammonium or by sodium or spermine, and TEA did not potentiate the activity of other inwardly rectifying potassium (K-ATP) channels in patches from cardiac myocytes. 6. We consider the possibility that TEA is mimicking the effect of an unidentified cellular factor, not sodium or PIP2, which normally occupies the TEA site on K-ACh channel proteins but which diffuses away when the patch is excised. C1 NIEHS, Lab Signal Transduct F205, Res Triangle Pk, NC 27709 USA. RP Armstrong, DL (reprint author), NIEHS, Lab Signal Transduct F205, POB 12233,111 Alexander Dr, Res Triangle Pk, NC 27709 USA. NR 30 TC 1 Z9 1 U1 0 U2 0 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0022-3751 J9 J PHYSIOL-LONDON JI J. Physiol.-London PD DEC 15 PY 2000 VL 529 IS 3 BP 699 EP 705 DI 10.1111/j.1469-7793.2000.00699.x PG 7 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 390QC UT WOS:000166307000015 PM 11118499 ER PT J AU Grabczyk, E Usdin, K AF Grabczyk, E Usdin, K TI Alleviating transcript insufficiency caused by Friedreich's ataxia triplet repeats SO NUCLEIC ACIDS RESEARCH LA English DT Article ID H-DNA; CLINICAL-FEATURES; SWITCH REGION; PLASMID DNA; SEQUENCES; EXPANSION; OLIGONUCLEOTIDE; FRATAXIN; TEMPLATE; BIOLOGY AB Expanded GAA.TTC trinucleotide repeats in intron 1 of the frataxin gene cause Friedreich's ataxia (FRDA) by reducing frataxin mRNA levels. Insufficient frataxin, a nuclear encoded mitochondrial protein, leads to the progressive neurodegeneration and cardiomyopathy characteristic of FRDA, Previously we demonstrated that long GAA.TTC tracts impede transcription elongation in vitro and provided evidence that the impediment results from an intramolecular purine.purine.pyrimidine DNA tripler formed behind an advancing RNA polymerase, Our model predicts that inhibiting formation of this tripler during transcription will increase successful elongation through GAA.TTC tracts, Here we show that this is the case, Oligodeoxyribonucleotides designed to block particular types of tripler formation provide specific and concentration-dependent increases in full-length transcript, In principle, therapeutic agents that selectively interfere with tripler formation could alleviate the frataxin transcript insufficiency caused by pathogenic FRDA alleles. C1 NIDDKD, Sect Genom Struct & Funct, Mol & Cellular Biol Lab, NIH, Bethesda, MD 20892 USA. RP Grabczyk, E (reprint author), NIDDKD, Sect Genom Struct & Funct, Mol & Cellular Biol Lab, NIH, Bldg 8,Room 202,8 Ctr Dr MSC 0830, Bethesda, MD 20892 USA. NR 38 TC 46 Z9 46 U1 1 U2 2 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0305-1048 J9 NUCLEIC ACIDS RES JI Nucleic Acids Res. PD DEC 15 PY 2000 VL 28 IS 24 BP 4930 EP 4937 DI 10.1093/nar/28.24.4930 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 384QJ UT WOS:000165956100014 PM 11121484 ER PT J AU Smith, RD Malley, JD Schechter, AN AF Smith, RD Malley, JD Schechter, AN TI Quantitative analysis of globin gene induction in single human erythroleukemic cells SO NUCLEIC ACIDS RESEARCH LA English DT Article ID HUMAN-LEUKEMIA-CELLS; FETAL HEMOGLOBIN; ERYTHROID-CELLS; EXPRESSION; HEMIN; LINE; TRANSCRIPTION; AMPLIFICATION; SYSTEM AB The mechanisms involved in the normal developmental regulation of globin gene expression, and the response to pharmacological agents that elevate fetal hemoglobin, may be expected to involve either changes in each cell or a selection process affecting subsets Of differentiating erythroid cells. To study these mechanisms we have developed assays to measure mRNA levels in single erythroid cells, The assay involved the use of globin-specific probes, with no detectable cross-reactivity, in real-time, fluorescence-based quantitative PCR (Q-PCR), We had previously used this Q-PCR method to measure globin mRNA levels in cultures of primary erythroid cells demonstrating that drugs like hydroxyurea, 5-azacytidine and butyric acid each yielded increases in gamma/(gamma + beta) mRNA ratios, with differential effects on beta -globin levels. We have now extended this approach to measure globin mRNA levels in single K562 cells, a human erythroleukemic cell line, with and without 30 muM hemin treatment. Hemin exposure increases total hemoglobin levels by similar to9-fold and total alpha-, epsilon- and gamma -globin mRNA levels by 1.5-2.3-fold. Single cell analyses showed initial wide distributions of each of the three individual globin mRNA levels with most cells having detectable but very low levels of each globin transcript, Hemin induction shifted the distributions to higher levels, with a tendency to residual left skewing as some cells remained with very low expression levels despite the effect of hemin in increasing expression in most of these low expressing cells. Thus transcriptional heterogeneity remains a crucial variable, even in this extensively used model of human erythroid biology, and clearly influences strongly the response to inducing agents, These methods may enable us to define better possible molecular and/or cellular models of globin gene modulation. C1 NIDDK, Biol Chem Lab, NIH, Bethesda, MD 20892 USA. NIH, Ctr Informat Technol, Computat Biosci & Engn Lab, Bethesda, MD 20892 USA. RP Schechter, AN (reprint author), NIDDK, Biol Chem Lab, NIH, Bldg 10,Room 9N307,10 Ctr Dr,MSC 1822, Bethesda, MD 20892 USA. OI Schechter, Alan N/0000-0002-5235-9408 NR 35 TC 19 Z9 20 U1 0 U2 1 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0305-1048 J9 NUCLEIC ACIDS RES JI Nucleic Acids Res. PD DEC 15 PY 2000 VL 28 IS 24 BP 4998 EP 5004 DI 10.1093/nar/28.24.4998 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 384QJ UT WOS:000165956100021 PM 11121491 ER PT J AU Apud, JA Egan, MF Wyatt, RJ AF Apud, JA Egan, MF Wyatt, RJ TI Effects of smoking during antipsychotic withdrawal in patients with chronic schizophrenia SO SCHIZOPHRENIA RESEARCH LA English DT Article DE antipsychotic; drug withdrawal; neuroleptic; schizophrenia; smoking ID CIGARETTE-SMOKING; NEUROLEPTIC WITHDRAWAL; DOPAMINE RELEASE; NICOTINE; PERFORMANCE; PSYCHOPATHOLOGY; NORMALIZATION; INPATIENTS; CAFFEINE; BEHAVIOR AB A number of studies have shown that patients with schizophrenia smoke mon than other psychiatric patients and more than the general population. Also, medicated schizophrenics who smoke present more positive symptoms of schizophrenia than non-smokers. The objective of the present study was to assess the effect of smoking on ratings of psychopathology for 30 days following discontinuation of antipsychotic medication. The subjects were 101 treatment-resistant patients with schizophrenia who had been admitted to the inpatient service of Neuroscience Research Hospital (NRH), National Institute of Mental Health, between 1982 and 1994 to undergo studies involving discontinuation of antipsychotic medication. Patients were rated independently on a daily basis on the 212-item Psychiatric Symptom Assessment Scale (PSAS), an extended version of the Brief Psychiatric Rating Scale (BPRS). At baseline, ratings for Verbal Positive, Paranoia and Loss of Function were higher in smokers (n=65) than nonsmokers (n=36), but a statistically significant difference was observed only for the Verbal Positive cluster. Analysis by gender revealed that male non-smokers had the lowest psychopathology ratings at baseline. There were no differences in Anxiety/depression, Behavior Positive, Deficit Symptoms or Mannerisms (a measure for abnormal involuntary movements). Following medication discontinuation, repeated-measure analysis demonstrated a 'time' effect for all the variables studied and a 'group' (smokers vs. Iron-smokers) effect for Verbal Positive. Paranoia, and Loss of Function. Post-hoc comparisons at individual time points showed significantly higher ratings for smokers at week 1 for Paranoia. No differences were observed at later time points. In conclusion, at baseline, smokers had more positive symptoms and were apparently more functionally impaired than non-smokers. This difference was no longer evident after a 30 day medication discontinuation period, (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIMH, St Elizabeths Hosp, Ctr Neurosci, Neuropsychiat Branch, Washington, DC 20032 USA. RP Apud, JA (reprint author), St Elizabeth Hosp, Psychopharmacol Div, Dept Psychiat, Barton Hall,2700 Martin Luther King Jr Ave SE, Washington, DC 20032 USA. NR 37 TC 7 Z9 7 U1 1 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0920-9964 J9 SCHIZOPHR RES JI Schizophr. Res. PD DEC 15 PY 2000 VL 46 IS 2-3 BP 119 EP 127 DI 10.1016/S0920-9964(99)00230-3 PG 9 WC Psychiatry SC Psychiatry GA 388VQ UT WOS:000166204100006 PM 11120424 ER PT J AU Elvevag, B Egan, MF Goldberg, TE AF Elvevag, B Egan, MF Goldberg, TE TI Memory for temporal order in patients with schizophrenia SO SCHIZOPHRENIA RESEARCH LA English DT Article DE memory; temporal order and schizophrenia ID MATCHED TASKS; RECALL; RECOGNITION; DYSFUNCTION; WORKING AB There is some evidence that memory for temporal order is a process that may be impaired independently of other forms of memory. For example. patients with Korsakoff's syndrome have been shown to have poorer temporal-order memory than other amnesic patients, despite item memory being equivalent. patients with schizophrenia have been reported to have a variety of memory problems, although memory for the order of events has not been examined very frequently. In this study, we tested memory for temporal order in patients with schizophrenia and in control subjects. Subjects were presented with two lists of 15 words (at two different times) and were Eater asked to reproduce the order of each list from a random array of the words. In both versions of the test, patients with schizophrenia were impaired in placing the items in the correct temporal order. Recall and recognition of the actual words used to comprise the lists were also impaired in the schizophrenic patients. However. when recall measures were covaried, and when patients were matched with controls for recall, post-hoc group differences in temporal memory were eliminated. In contrast, covarying recognition (indexed by d' or matching for recognition) did not eliminate group differences. Therefore, although memory for temporal order is compromised in patients with schizophrenia. this deficit is highly correlated with generally poorer item-specific memory retrieval (i.e.. recall). It is possible that both impairments are due to some third process that underlies and aids in the reconstruction of episodes. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIMH, Clin Brain Disorders Branch, NIH, Bethesda, MD 20892 USA. RP Elvevag, B (reprint author), NIMH, Clin Brain Disorders Branch, NIH, Bethesda, MD 20892 USA. NR 28 TC 24 Z9 24 U1 1 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0920-9964 J9 SCHIZOPHR RES JI Schizophr. Res. PD DEC 15 PY 2000 VL 46 IS 2-3 BP 187 EP 193 DI 10.1016/S0920-9964(00)00014-1 PG 7 WC Psychiatry SC Psychiatry GA 388VQ UT WOS:000166204100012 PM 11120430 ER PT J AU Liu, XC Sun, ZX Uchiyama, M Shibui, K Kim, K Okawa, M AF Liu, XC Sun, ZX Uchiyama, M Shibui, K Kim, K Okawa, M TI Prevalence and correlates of sleep problems in Chinese schoolchildren SO SLEEP LA English DT Article DE child; sleep problems; dyssomnias; parasomnias; hyperactivity; correlates ID ATTENTION-DEFICIT/HYPERACTIVITY DISORDER; SCHOOL-CHILDREN; BEHAVIOR PROBLEMS; DISTURBANCES; AREA; RISK; AGE AB Study Objectives: This study examined the prevalence and correlates of sleep problems in Chinese schoolchildren, Design and Setting: A cross-sectional questionnaire survey was undertaken in Shandong Province, People's Republic of China. Participants: A total of 2004 elementary school children (998 boys and 1006 girls) participated in the survey Measurements and Interventions: The parents completed a questionnaire that asked about sleep problems, and characteristics of the family and child. Teachers completed a questionnaire that included the Modified Conners Hyperkinesis Index (MCHI), whether the child slept in class, and school achievement. Results: Parent-reported sleep problems that occurred "sometimes" or "often" were sleep walking/talking, 14.2%; too little sleep,. 14.0%; too much sleep, 12.5%; nightmares, 12.0%; trouble sleeping, 6.1%; and nocturnal enuresis, 4.5%. Teachers reported that 9.4% of children slept in class "sometimes" or "often". Approximately 11% of children were reported to have any sleep problem "often". Children with sleep problems were more frequently reported to be hyperactive, and to have poorer child-parent relations, poorer peer relations, and poorer social competency and school achievement. Multivariate logistic regression analysis indicated that sleep problems were significantly correlated with following factors: poor parental relations, crowded homes, bedwetting cessation after age 4, chronic physical diseases, reported hyperactivity and poor peer relations. Conclusions: Parent-reported sleep problems in Chinese children were less prevalent than those reported in Western countries, and associated with multiple family, prenatal, and child developmental factors. Children with sleep problems were reported to be more hyperactive, and to have social and academic problems more frequently. C1 Shandong Med Univ, Dept Psychiat, Shandong, Peoples R China. Natl Inst Mental Hlth, Dept Psychophysiol, Chiba, Japan. RP Liu, XC (reprint author), NIEHS, Epidemiol Branch, Mail Drop A3-05,POB 12233,111 TW Alexander Dr,Roo, Res Triangle Pk, NC 27709 USA. NR 43 TC 55 Z9 59 U1 1 U2 9 PU AMER SLEEP DISORDERS ASSOC PI ROCHESTER PA 6301 BANDEL RD, STE 101, ROCHESTER, MN 55901 USA SN 0161-8105 J9 SLEEP JI Sleep PD DEC 15 PY 2000 VL 23 IS 8 BP 1053 EP 1062 PG 10 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 384XU UT WOS:000165973200007 PM 11145320 ER PT J AU Smith, TD Kuczenski, R George-Friedman, K Malley, JD Foote, SL AF Smith, TD Kuczenski, R George-Friedman, K Malley, JD Foote, SL TI In vivo microdialysis assessment of extracellular serotonin and dopamine levels in awake monkeys during sustained fluoxetine administration SO SYNAPSE LA English DT Article DE nonhuman primate; depression; antidepressant; Prozac; hippocampus; neocortex; caudate putamen ID RAT-BRAIN; PRIMATE NEOCORTEX; CEREBRAL-CORTEX; AXON TERMINALS; IN-VIVO; DORSAL; INCREASE; 5-HT1A; AUTORECEPTORS; PROJECTIONS AB Fluoxetine (FLU) rapidly enhances extracellular (EC) serotonin (5-HT) in rodent brain, whereas the antidepressant effects of this drug in humans are typically not observed for 2-3 weeks. Thus, the effects of chronic oral FLU administration on neocortical and hippocampal EC 5-HT, and on caudate EC 5-HT and dopamine (DA), were examined in awake monkeys (Macaca fascicularis) using in vivo microdialysis (10.0 mg/kg; 3, 7, 14, and 21 days). On day 3, 5-HT was significantly increased above baseline levels in hippocampus (HC) and caudate. There was a trend for an increase in neocortex EC 5-HT levels. However, by day 7 5-HT remained significantly elevated only in KC, although 5-HT levels elsewhere had not completely returned to baseline, in contrast, levels of the 5-HT metabolite, 5-HIAA, were significantly reduced in all brain regions at all time points. Caudate DA levels tended to be decreased throughout FLU treatment. Local FLU and K+ infusion were also used at various times during chronic systemic FLU administration to evaluate changes in functional synaptic regulation. In general, these results, along with the significant decrease in 5-HIAA levels and the tendency for basal EC 5-HT levels to remain modestly elevated only in HC during sustained FLU administration, suggest a reduction in releasable pools of 5-HT. Taken together with the trend for a decrease in caudate EC DA levels, these results do not appear to support the current hypothesis regarding the mechanism of action of SSRI antidepressants - that monoaminergic neurotransmission is progressively augmented during chronic treatment. (C) 2000 Wiley-Liss, Inc. C1 Univ Calif San Diego, Sch Med, Dept Psychiat, La Jolla, CA 92093 USA. NIH, Ctr Informat Technol, Bethesda, MD 20892 USA. NIMH, Bethesda, MD 20892 USA. RP Kuczenski, R (reprint author), Univ Calif San Diego, Sch Med, Dept Psychiat, 9500 Gilman Dr,Box 0603, La Jolla, CA 92093 USA. FU NIMH NIH HHS [MH40008] NR 33 TC 39 Z9 40 U1 1 U2 3 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0887-4476 J9 SYNAPSE JI Synapse PD DEC 15 PY 2000 VL 38 IS 4 BP 460 EP 470 DI 10.1002/1098-2396(20001215)38:4<460::AID-SYN11>3.0.CO;2-D PG 11 WC Neurosciences SC Neurosciences & Neurology GA 370XC UT WOS:000165147700011 PM 11044893 ER PT J AU Delon, J Germain, RN AF Delon, J Germain, RN TI Information transfer at the immunological synapse SO CURRENT BIOLOGY LA English DT Review ID T-CELL-RECEPTOR; PEPTIDE-MHC COMPLEXES; MICROTUBULE-ORGANIZING CENTER; PRESENTING B-CELLS; MEMBRANE-ASSOCIATED CYTOSKELETON; CLASS-I PROTEINS; SIGNAL-TRANSDUCTION; MINIMAL NUMBER; ANTIGEN RECOGNITION; ACTIN CYTOSKELETON AB Antigen specific activation of T lymphocytes requires the interaction of their clonally distributed T-cell receptors with plasma membrane ligands composed of foreign peptide antigens bound to major histocompatibility complex molecules. For proliferation and differentiation to ensue, a variety of other adhesive and accessory proteins must also interact with their counter-receptors on the antigen-presenting cell to facilitate and complement the T-cell receptor-antigen recognition event. Recent studies have revealed that these various proteins show an unexpected degree of spatial organization in the zone of cell-cell contact. This region of membrane approximation is now referred to as the 'immunological synapse' because of its functional analogy to the site of intercellular information transfer between neurons. Here, we review the evidence for signaling-dependent control of the dynamic changes in protein distribution that gives rise to the synapse and try to relate the emerging spatio-temporal information on synapse formation to T-cell biology. C1 NIAID, Lymphocyte Biol Sect, Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Germain, RN (reprint author), NIAID, Lymphocyte Biol Sect, Immunol Lab, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 106 TC 57 Z9 60 U1 0 U2 0 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0960-9822 J9 CURR BIOL JI Curr. Biol. PD DEC 14 PY 2000 VL 10 IS 24 BP R923 EP R933 DI 10.1016/S0960-9822(00)00870-8 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 399JD UT WOS:000166807100013 PM 11137031 ER PT J AU Peculis, BA AF Peculis, BA TI RNA-binding proteins: If it looks like a sn(o)RNA ... SO CURRENT BIOLOGY LA English DT Article ID TRI-SNRNP; IDENTIFICATION AB Small nuclear RNAs are involved in splicing pre mRNA, while small nucleolar RNAs facilitate ribosome biogenesis. But these distinct particles may have more in common than was first apparent: some of their RNA components share a common RNA binding protein, a common RNA structure and perhaps a common origin. C1 NIDDK, Genet & Biochem Branch, NIH, Bethesda, MD 20892 USA. RP Peculis, BA (reprint author), NIDDK, Genet & Biochem Branch, NIH, Bethesda, MD 20892 USA. NR 7 TC 6 Z9 6 U1 0 U2 0 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0960-9822 J9 CURR BIOL JI Curr. Biol. PD DEC 14 PY 2000 VL 10 IS 24 BP R916 EP R918 DI 10.1016/S0960-9822(00)00851-4 PG 3 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 399JD UT WOS:000166807100011 PM 11137029 ER PT J AU Zhang, Y Rothman, RB Dersch, CM de Costa, BR Jacobson, AE Rice, KC AF Zhang, Y Rothman, RB Dersch, CM de Costa, BR Jacobson, AE Rice, KC TI Synthesis and transporter binding properties of bridged piperazine analogues of 1-{2-[bis(4-fluorophenyl)methoxy]ethyl}-4-(3-phenylpropyl)piperazine (GBR 12909) SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID DOPAMINE REUPTAKE INHIBITORS; 1-<2-ETHYL>-4-(3-PHENYLPROPYL)PIPERAZINES GBR-12935; NOREPINEPHRINE TRANSPORTER; EXTRACELLULAR DOPAMINE; NONHUMAN-PRIMATES; ANIMAL-MODELS; COCAINE-ABUSE; KNOCKOUT MICE; AMPHETAMINE; RAT AB A series of analogues related to 1-[2-(diphenylmethoxy)ethyl]-4-(3-phenylpropyl)piperazine (2) and 1-{2-[bis(4-fluorophenyl)methoxy]ethyl)-4-(3-phenylpropyl)piperazine (3)(GBR 12935 and GBR 12909, respectively), in which;the piperazine moiety was replaced by bridged piperazines for structural rigidity,has been designed, synthesized, and evaluated for their ability to bind to the dopamine transporter;(DAT) and to inhibit the uptake of H-3-labeled dopamine (DA), The binding data indicated that compounds 7 and Il, the N-methyl- and N-propylphenyl-3,8-diaza[3.2.1]bicyclooctane analogues of 3, showed high affinity for the DAT (IC50 = 8.0 and 8.2 nM, respectively), and 7 had,high selectivity at the DAT relative to the serotonin transported (SERT) (88- and 93-fold for binding and reuptake, respectively). They also displayed linear activity in DA uptake inhibition, possessing a similar binding and reuptake inhibition profile to 3. The N-indolylmethyl analogue;16 showed the highest affinity (IC50 = 1.4 nM) of the series, with a 6-fold increase over its :corresponding N-phenypropyl derivative II. Interestingly, this compound exhibited a high ratio (29-fold) of IC50 for the inhibition of DA reuptake versus binding to the DAT.; Replacing the piperazine moiety of 2 and 3 with (1S,4S)-2,5-diazabicyclo [2.2.1]-heptane resulted in compounds 23-26, which showed moderate to poor affinity (IC50 = 127-1170 nM)for the;DAT: Substitution of the homopiperazine moiety of 4 with a more rigid 3,9-diazabicyclo [4.2.1]nonane gave compounds 28-33. However, the binding data showed that compound 32 displayed a 10-fold decrease in affinity at the DAT and a 100-fold decrease in selectivity at the DAT relative to the SERT compared to its corresponding homopiperazine compound 4. C1 NIDDK, Med Chem Lab, NIH, Bethesda, MD 20892 USA. NIAID, Clin Psychopharmacol Sect, Addict Res Ctr, Baltimore, MD 21224 USA. RP Rice, KC (reprint author), NIDDK, Med Chem Lab, NIH, Bldg 8,Rm B1-23,8 Ctr Dr,MSC 0815, Bethesda, MD 20892 USA. NR 55 TC 35 Z9 35 U1 3 U2 3 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0022-2623 J9 J MED CHEM JI J. Med. Chem. PD DEC 14 PY 2000 VL 43 IS 25 BP 4840 EP 4849 DI 10.1021/jm000300r PG 10 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 383CT UT WOS:000165866100014 PM 11123994 ER PT J AU Gu, XH Yu, H Jacobson, AE Rothman, RB Dersch, CM George, C Flippen-Anderson, JL Rice, KC AF Gu, XH Yu, H Jacobson, AE Rothman, RB Dersch, CM George, C Flippen-Anderson, JL Rice, KC TI Design, synthesis, and monoamine transporter binding site affinities of methoxy derivatives of indatraline SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID RHESUS-MONKEYS; DOPAMINE; SEROTONIN; METHAMPHETAMINE; ACIDS; NOREPINEPHRINE; INHIBITION; GBR-12909; GBR12909; REMOVAL AB A series of methoxy-containing derivatives of indatraline 13a-f and 17 were synthesized, and their binding affinities for the dopamine, serotonin, and norepinephrine transporter binding sites were determined. Introduction of a methoxy group to indatraline affected its affinity and selectivity greatly. Except for the 4-methoxy derivative 13a,which had the same high affinity at the dopamine transporter binding site as indatraline, the other methoxy-containing analogues (13b-f and 17) exhibited lower affinity than indatraline for the three transporter binding sites. However, some of the analogues were more selective than indatraline, and the B-methoxy derivative 13c displayed the highest affinity for both the serotonin and norepinephrine transporters. This compound retained reasonable affinity for the dopamine transporter and is a promising template for the development of a long-acting inhibitor of monoamine transporters. Such inhibitors have potential as medications for treatment, as a substitution medication, or for prevention of the abuse of methamphetamine-like stimulants. C1 NIDDKD, Med Chem Lab, NIH, Bethesda, MD 20892 USA. NIDA, Clin Psychopharmacol Sect, Baltimore, MD 21224 USA. USN, Res Lab, Struct Matter Lab, Washington, DC 20375 USA. RP Rice, KC (reprint author), NIDDKD, Med Chem Lab, NIH, Bldg 8,Room B1-23, Bethesda, MD 20892 USA. NR 35 TC 31 Z9 32 U1 0 U2 1 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0022-2623 J9 J MED CHEM JI J. Med. Chem. PD DEC 14 PY 2000 VL 43 IS 25 BP 4868 EP 4876 DI 10.1021/jm000329v PG 9 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 383CT UT WOS:000165866100016 PM 11123996 ER PT J AU Misteli, T Gunjan, A Hock, R Bustin, M Brown, DT AF Misteli, T Gunjan, A Hock, R Bustin, M Brown, DT TI Dynamic binding of histone H1 to chromatin in living cells SO NATURE LA English DT Article ID HIGHER-ORDER STRUCTURE; GENE-EXPRESSION; IN-VIVO; ACETYLATION; TRANSCRIPTION; PHOSPHORYLATION; PROMOTER; EXCHANGE; CYCLE; BASAL AB The linker histone H1 is believed to be involved in chromatin organization by stabilizing higher-order chromatin structure(1-3). Histone H1 is generally viewed as a repressor of transcription as it prevents the access of transcription factors and chromatin remodelling complexes to DNA(4-6). Determining the binding properties of histone H1 to chromatin in vivo is central to understanding how it exerts these functions. We have used photobleaching techniques to measure the dynamic binding of histone H1-GFP to unperturbed chromatin in living cells. Here we show that almost the entire population of H1-GFP is bound to chromatin at any one time; however, H1-GFP is exchanged continuously between chromatin regions. The residence time of H1-GFP on chromatin between exchange events is several minutes in both euchromatin and heterochromatin. In addition to the mobile fraction, we detected a kinetically distinct, less mobile fraction. After hyperacetylation of core histones, the residence time of H1-GFP is reduced, suggesting a higher rate of exchange upon chromatin remodelling. These results support a model in which linker histones bind dynamically to chromatin in a stop-and-go mode. C1 NCI, NIH, Bethesda, MD 20892 USA. Univ Mississippi, Med Ctr, Jackson, MS 39216 USA. Univ Wurzburg, Bioctr, D-97070 Wurzburg, Germany. NCI, NIH, Mol Biol Lab, Bethesda, MD 20892 USA. RP Misteli, T (reprint author), NCI, NIH, Bethesda, MD 20892 USA. RI Bustin, Michael/G-6155-2015 NR 29 TC 412 Z9 419 U1 1 U2 20 PU MACMILLAN PUBLISHERS LTD PI LONDON PA PORTERS SOUTH, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD DEC 14 PY 2000 VL 408 IS 6814 BP 877 EP 881 DI 10.1038/35048610 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 382PK UT WOS:000165831300055 PM 11130729 ER PT J AU Rai, KR Peterson, BL Appelbaum, FR Kolitz, J Elias, L Shepherd, L Hines, J Threatte, GA Larson, RA Cheson, BD Schiffer, CA AF Rai, KR Peterson, BL Appelbaum, FR Kolitz, J Elias, L Shepherd, L Hines, J Threatte, GA Larson, RA Cheson, BD Schiffer, CA TI Fludarabine compared with chlorambucil as primary therapy for chronic lymphocytic leukemia. SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Article; Proceedings Paper CT 38th Annual Meeting of the American-Society-of-Hematology CY DEC 06-10, 1996 CL ORLANDO, FL SP Amer Soc Hematol, US Dept Vet Affairs Merit Res Funds, NIH ID TERM FOLLOW-UP; CLINICAL-TRIALS; AGENT; PREDNISONE AB Background: Fludarabine is an effective treatment for chronic lymphocytic leukemia that does not respond to initial treatment with chlorambucil. We compared the efficacy of fludarabine with that of chlorambucil in the primary treatment of chronic lymphocytic leukemia. Methods: Between 1990 and 1994, we randomly assigned 509 previously untreated patients with chronic lymphocytic leukemia to one of the following treatments: fludarabine (25 mg per square meter of body-surface area, administered intravenously daily for 5 days every 28 days), chlorambucil (40 mg per square meter, given orally every 28 days), or fludarabine (20 mg per square meter per day for 5 days every 28 days) plus chlorambucil (20 mg per square meter every 28 days). Patients with an additional response at each monthly evaluation continued to receive the assigned treatment for a maximum of 12 cycles. Results: Assignment of patients to the fludarabine-plus-chlorambucil group was stopped when a planned interim analysis revealed excessive toxicity and a response rate that was not better than the rate with fludarabine alone. Among the other two groups, the response rate was significantly higher for fludarabine alone than for chlorambucil alone. Among 170 patients treated with fludarabine, 20 percent had a complete remission, and 43 percent had a partial remission. The corresponding values for 181 patients treated with chlorambucil were 4 percent and 33 percent (P<0.001 for both comparisons). The median duration of remission and the median progression-free survival in the fludarabine group were 25 months and 20 months, respectively, whereas both values were 14 months in the chlorambucil group (P<0.001 for both comparisons). The median overall survival among patients treated with fludarabine was 66 months, which was not significantly different from the overall survival in the other two groups (56 months with chlorambucil and 55 months with combined treatment). Severe infections and neutropenia were more frequent with fludarabine than with chlorambucil (P=0.08), although, overall, toxic effects were tolerable with the two single-drug regimens. Conclusions: When used as the initial treatment for chronic lymphocytic leukemia, fludarabine yields higher response rates and a longer duration of remission and progression-free survival than chlorambucil; overall survival is not enhanced. (N Engl J Med 2000;343:1750-7.) (C) 2000, Massachusetts Medical Society. C1 Canc Leukemia Grp B, Chicago, IL USA. SW Oncol Grp, San Antonio, TX 78229 USA. Natl Canc Inst, Clin Trials Grp, Kingston, ON, Canada. Eastern Cooperat Oncol Grp, Brookline, MA USA. NCI, Rockville, MD USA. RP Long Isl Jewish Med Ctr, 270-05 76th Ave, New Hyde Pk, NY 11040 USA. EM rai@lij.edu OI Larson, Richard/0000-0001-9168-3203 FU NCI NIH HHS [CA21115, CA31946, CA32102] NR 17 TC 642 Z9 667 U1 2 U2 21 PU MASSACHUSETTS MEDICAL SOC PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 EI 1533-4406 J9 NEW ENGL J MED JI N. Engl. J. Med. PD DEC 14 PY 2000 VL 343 IS 24 BP 1750 EP 1757 DI 10.1056/NEJM200012143432402 PG 8 WC Medicine, General & Internal SC General & Internal Medicine GA 382FH UT WOS:000165812000002 PM 11114313 ER PT J AU Benvenuto, G Li, SW Brown, SJ Braverman, R Vass, WC Cheadle, JP Halley, DJJ Sampson, JR Wienecke, R DeClue, JE AF Benvenuto, G Li, SW Brown, SJ Braverman, R Vass, WC Cheadle, JP Halley, DJJ Sampson, JR Wienecke, R DeClue, JE TI The tuberous sclerosis-1 (TSC1) gene product hamartin suppresses cell growth and augments the expression of the TSC2 product tuberin by inhibiting its ubiquitination SO ONCOGENE LA English DT Article DE tuberous sclerosis (TSC); tumor suppressor genes; growth inhibition; ubiquitination ID NORMAL HUMAN TISSUES; EKER RAT; NEOPLASTIC PHENOTYPE; PROTEASOME PATHWAY; RENAL-CARCINOMA; HUMAN CANCER; TARGET RAP1; IN-VIVO; PROTEIN; P53 AB We report here that overexpression of the tuberous sclerosis-1 (TSC1) gene product hamartin results in the inhibition of growth, as well as changes in cell morphology, Growth inhibition was associated with an increase in the endogenous level of the product of the tuberous sclerosis-2 (TSC2) gene, tuberin. As over-expression of tuberin inhibits cell growth, and hamartin is known to bind tuberin, these results suggested that hamartin stabilizes tuberin and this contributes to the inhibition of cell growth. Indeed, transient transfection of TSC1 increased the endogenous level of tuberin, and transient co-transfection of TSC1 with TSC2 resulted in higher tuberin levels. The stabilization was explained by the finding that tuberin is highly ubiquitinated in cells, while the fraction of tuberin that is bound to hamartin is not ubiquitinated, Go-expression of tuberin stabilized hamartin, which is weakly ubiquitinated, in transiently transfected cells. The amino-terminal two-thirds of tuberin was responsible for its ubiquitination and for stabilization of hamartin, A mutant of tuberin from a patient missense mutation of TSC2 was also highly ubiquitinated, and was unable to stabilize hamartin, We conclude that hamartin is a growth inhibitory protein whose biological effect is likely dependent on its interaction with tuberin. C1 NCI, Cellular Oncol Lab, Bethesda, MD 20892 USA. Univ Wales Coll Med, Inst Med Genet, Cardiff CF4 4XN, S Glam, Wales. Erasmus Univ, Dept Clin Genet, NL-3015 GE Rotterdam, Netherlands. Univ Munich, Dept Dermatol, Klinikum Innenstadt, D-80337 Munich, Germany. RP DeClue, JE (reprint author), NCI, Cellular Oncol Lab, Bethesda, MD 20892 USA. NR 42 TC 141 Z9 144 U1 1 U2 5 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD DEC 14 PY 2000 VL 19 IS 54 BP 6306 EP 6316 DI 10.1038/sj.onc.1204009 PG 11 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 388YK UT WOS:000166210500014 PM 11175345 ER PT J AU Gudi, T Casteel, DE Vinson, C Boss, GR Pilz, RB AF Gudi, T Casteel, DE Vinson, C Boss, GR Pilz, RB TI NO activation of fos promoter elements requires nuclear translocation of G-kinase I and CREB phosphorylation but is independent of MAP kinase activation SO ONCOGENE LA English DT Article DE nitric oxide; cGMP-dependent protein kinase; c-fos; CREB ID DEPENDENT PROTEIN-KINASE; MULTIPLE SEQUENCE ELEMENTS; SOLUBLE GUANYLATE-CYCLASE; SMOOTH-MUSCLE CELLS; NITRIC-OXIDE; CYCLIC-GMP; C-FOS; GENE-EXPRESSION; BINDING PROTEIN; DNA-BINDING AB We have shown that nitric oxide (NO) regulates c-fos gene expression via cGMP-dependent protein kinase (G-kinase), but NO's precise mechanism of action is unclear, We now demonstrate that: (1) NO targets two transcriptional elements in the fos promoter, i,e., the fos AP-1 binding site and the cAMP-response element (CRE); (2) NO activation of these two enhancer elements requires the CRE binding protein CREB because a dominant negative CREB fully inhibits NO transactivation of reporter genes whereas dominant negative Fos or CCAAT enhancer binding proteins have no effect; (3) CREB is phosphorylated by G-kinase in vitro and its phosphorylation increases in vivo when G-kinase is activated either directly by cGMP or indirectly by NO via soluble guanylate cyclase; (4) NO activation of fos promoter elements requires nuclear translocation of G-kinase but not activation of mitogen-activated protein kinases. C1 Univ Calif San Diego, Dept Med, La Jolla, CA 92093 USA. Univ Calif San Diego, Ctr Canc, La Jolla, CA 92093 USA. NCI, Met Lab, NIH, Bethesda, MD 20892 USA. RP Pilz, RB (reprint author), Univ Calif San Diego, Dept Med, La Jolla, CA 92093 USA. FU NIDDK NIH HHS [5T32DK07233-24]; NIGMS NIH HHS [R01GM055586] NR 61 TC 67 Z9 72 U1 0 U2 1 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD DEC 14 PY 2000 VL 19 IS 54 BP 6324 EP 6333 DI 10.1038/sj.onc.1204007 PG 10 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 388YK UT WOS:000166210500016 PM 11175347 ER PT J AU Donini, O Darden, T Kollman, PA AF Donini, O Darden, T Kollman, PA TI QM-FE calculations of aliphatic hydrogen abstraction in citrate synthase and in solution: Reproduction of the effect of enzyme catalysis and demonstration that an enolate rather than an enol is formed SO JOURNAL OF THE AMERICAN CHEMICAL SOCIETY LA English DT Article ID FREE-ENERGY CALCULATIONS; TRANSITION-STATES; CARBON ACIDS; ACETYL-COA; TRIOSEPHOSPHATE ISOMERASE; ELECTROPHILIC CATALYSIS; MOLECULAR-DYNAMICS; PROTON ABSTRACTION; SERINE PROTEASES; AQUEOUS-SOLUTION AB Mechanistic enzymologists have long debated how enzymes catalyze the abstraction of an unactivated C-H group. Citrate synthase, due to its ability to catalyze this abstraction and its central role in the respiratory cycle, has been extensively studied both experimentally and theoretically. Despite this scrutiny, the question remains as to whether the initial aliphatic hydrogen abstraction step of the mechanism is stabilized by the formation of an enol-imidazolate intermediate through "short, strong" hydrogen bonds, as opposed to the more traditional enolate-imidazole complex. Tn an attempt to present a definitive answer to this question, quantum mechanical-free energy (QM-FE) calculations were performed for the formation of the enolate-imidazole complex from the reactants, as well as for the further formation of the enol-imidazolate system. These reactions were found to be extremely sensitive to the use of nonbonded cutoffs, and reliable results were only obtained with the use of particle mesh Ewald (PME) to heat the electrostatic interactions. Because of the length of these simulations, we also used a coarse-grained parallel approach to free energy calculations. The results indicate that the enolate-imidazole complex is the more stable one within the enzyme by approximately 13 kcal/mol. The calculated barrier to the formation of the enolate is in good quantitative agreement with the k(cat) for this enzyme. C1 Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94143 USA. NIEHS, Struct Biol Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Kollman, PA (reprint author), Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94143 USA. NR 51 TC 30 Z9 31 U1 0 U2 4 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0002-7863 J9 J AM CHEM SOC JI J. Am. Chem. Soc. PD DEC 13 PY 2000 VL 122 IS 49 BP 12270 EP 12280 DI 10.1021/ja001043i PG 11 WC Chemistry, Multidisciplinary SC Chemistry GA 383CW UT WOS:000165866400024 ER PT J AU Speir, E Yu, ZX Takeda, K Ferrans, VJ Cannon, RO AF Speir, E Yu, ZX Takeda, K Ferrans, VJ Cannon, RO TI Antioxidant effect of estrogen on cytomegalovirus-induced gene expression in coronary artery smooth muscle cells SO CIRCULATION LA English DT Article DE atherosclerosis; viruses hormones; antioxidants; adhesion molecules ID PROTEIN-KINASE-C; ENDOTHELIAL-CELLS; OXIDATIVE STRESS; TAMOXIFEN; TRANSCRIPTION; INCREASES; ESTRADIOL; INHIBITION; DISEASE; THERAPY AB Background-Pathogens infecting the arterial wall with resultant inflammation may contribute to atherogenesis. Human coronary artery smooth muscle cells (SMCs) infected with human cytomegalovirus (CMV) demonstrate a rapid increase in reactive oxygen species (ROSs), with activation of genes involved in viral replication and inflammation. Because estrogen appears to have antioxidant properties, we wished to determine whether this hormone attenuates SMC responses to CMV infection. Methods and Results-Using confocal microscopy and an intracellular fluorescent dye activated by ROSs, we found that 17 beta -estradiol (0.1 to 10 nmol/L) and its stereoisomer 17 alpha -estradiol (which has low affinity for the estrogen receptor) dose-dependently inhibited ROS generation in CMV-infected SMCs. These effects were not blacked by the estrogen receptor inhibitor ICI 182,780. 3-Methoxyestrone, which lacks the phenolic hydroxyl group, did not interfere with ROS generation. We found that 17 beta -estradiol and 17 alpha -estradiol, but not 3-methoxyestrone, prevented binding of nuclear factor (NF)-kappaB to DNA. Furthermore, in SMCs transfected with the reporter constructs 3X kappaB-CAT, MIEP-CAT, or ICAM-CAT, cotransfection with a CMV-IE72 expression plasmid caused promoter and CAT activation. Treatment with 17 beta -estradiol and 17 alpha -estradiol, but not 3-methoxyestrone, inhibited CAT activity and, in CMV-infected SMCs, prevented IE72 and ICAM-1 protein expression and cytopathic effects. Conclusions-These findings indicate that estrogen molecules with an A-ring hydroxyl group have estrogen receptor-independent anti-CMV effects at physiological concentrations by inhibiting ROS generation, NF-kappaB activation, NF-kappaB-dependent transcription, and viral replication. To the extent that chronic infection of the vascular wall with CMV contributes to atherogenesis, these antioxidant actions of estrogen may be of therapeutic importance. C1 NHLBI, Cardiol Branch, NIH, Bethesda, MD 20892 USA. NHLBI, Pathol Sect, NIH, Bethesda, MD 20892 USA. RP Speir, E (reprint author), NHLBI, Cardiol Branch, NIH, Bldg 10,Room 7B15,10 Ctr Dr MSC-1650, Bethesda, MD 20892 USA. NR 28 TC 43 Z9 44 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD DEC 12 PY 2000 VL 102 IS 24 BP 2990 EP 2996 PG 7 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 382GA UT WOS:000165813600021 PM 11113051 ER PT J AU Fedorova, OV Lakatta, EG Bagrov, AY AF Fedorova, OV Lakatta, EG Bagrov, AY TI Endogenous Na,K pump ligands are differentially regulated during acute NaCl loading of Dahl rats SO CIRCULATION LA English DT Article; Proceedings Paper CT 71st Scientific Session of the American-Heart-Association Meeting CY NOV 08-12, 1998 CL DALLAS, TEXAS SP Amer Heart Assoc DE hypertension; rats, inbred Dahl; sodium, dietary; Na(+)-K(+)-exchanging ATPase; steroids; bufanolides; ouabain ID SALT-RESISTANT RATS; MARINOBUFAGENIN-LIKE; VOLUME EXPANSION; ALPHA-SUBUNIT; OUABAIN; SODIUM; PLASMA; HYPERTENSION; NA,K-ATPASE; ATPASE AB Background-Two mammalian digitalis-like factors, an ouabain-like compound (OLC) and marinobufagenin (MBG), exhibit specificity to alpha -3 and alpha -1 Na+, K+-ATPase isoforms, respectively. We compared regulation of MBG and OLC by acute NaCl loading in Dahl salt-sensitive (DS) and salt-resistant (DR) rats. Methods and Results-An intraperitoneal NaCl load (0.8 g/kg) was given to adult male rats (24 DS and 24 DR). Diuresis, natriuresis, renal excretion, and tissue levels of MBG and OLC were measured. Inhibition of renal Na+,K+-ATPase by MBC and ouabain was compared in DS, DR, and Wistar rats. DS (versus DR) exhibited a smaller peak (2 hours) natriuretic response (1.34+/-0.10 versus 2.08+/-0.14 mmol.kg(-1.)h(-1); P<0.01), despite a greater plasma Na+ (153+/-2 versus 145+/-1 mmol/L; P<0.01). In DS and DR, pituitary, adrenal, and plasma OLC exhibited transient 2-fold to 3-fold increases, followed by a decrease to baseline levels. Plasma and adrenal MBG doubled in both strains within 1 hour of NaCl loading and remained elevated. Eight-hour MBG excretion in DS was 4-fold greater than in DR (15.8+/-0.8 versus 3.6+/-0.4 pmol; P<0.01), whereas OLC excretion in DS was only 30% greater than in DR (16.1+/-1.1 and 11.9+/-0.8 pmol; P<0.05). Kidney Na+,K+-ATPase (alpha -1 isoform) from Wistar rats and DS exhibited greater sensitivity to MBC than to ouabain. Conclusions-NaCl loading of DS causes transient increase in OLC but sustained increases in MBG tissue levels and excretion. We hypothesize that increased MBG production occurs in an attempt to compensate for genetically impaired pressure-natriuresis mechanisms. C1 NIA, Cardiovasc Sci Lab, Intramural Res Program, Baltimore, MD 21224 USA. RP Bagrov, AY (reprint author), NIA, Cardiovasc Sci Lab, Intramural Res Program, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 28 TC 82 Z9 86 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD DEC 12 PY 2000 VL 102 IS 24 BP 3009 EP 3014 PG 6 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 382GA UT WOS:000165813600024 PM 11113054 ER PT J AU Kirschner, LS Sandrini, F Monbo, J Lin, JP Carney, JA Stratakis, CA AF Kirschner, LS Sandrini, F Monbo, J Lin, JP Carney, JA Stratakis, CA TI Genetic heterogeneity and spectrum of mutations of the PRKAR1A gene in patients with the Carney complex SO HUMAN MOLECULAR GENETICS LA English DT Article ID PSAMMOMATOUS MELANOTIC SCHWANNOMA; SPOTTY SKIN PIGMENTATION; SUBUNIT-RI-ALPHA; ENDOCRINE OVERACTIVITY; REGULATORY SUBUNIT; CARDIAC MYXOMAS; DUCTAL ADENOMA; PROTEIN-KINASE; ATRIAL-MYXOMA; HUMAN GENOME AB Carney complex (CNC) is an autosomal dominant multiple neoplasia syndrome, which has been linked to loci on 2p16 and 17q22-24. We recently reported that PRKAR1A, which codes for the type 1A regulatory subunit of protein kinase A (PKA), is a tumor suppressor gene on chromosome 17 that is mutated in some CNC families. To evaluate the spectrum of PRKAR1A mutations, we identified its genomic structure and screened for mutations in 54 CNC kindreds (34 families and 20 patients with sporadic disease). Fourteen families were informative for linkage analysis: four of four families that mapped to 17q had PRKAR1A mutations, whereas there were no mutations found in seven families exhibiting at least one recombination with 17q. In six of the latter, CNC mapped to 2p16, PRKAR1A mutations were also found in 12 of 20 non-informative families and 7 of 20 sporadic cases. Altogether, 15 distinct PRKAR1A mutations were identified in 22 of 54 kindreds (40.7%). In 14 mutations, the sequence change was predicted to lead to a premature stop codon; one altered the initiator ATG codon. Mutant mRNAs containing a premature stop codon were unstable, as a result of nonsense-mediated mRNA decay. Accordingly, the predicted truncated PRKAR1A protein products were absent in these cells. We conclude that (i) genetic heterogeneity exists in CNC; and (ii) all of the CNC alleles on 17q are functionally null mutations of PRKAR1A. CNC is the first human disease recognized to be caused by mutations of the PKA holoenzyme, a critical component of cellular signaling. C1 NICHHD, Unit Genet & Endocrinol, Dev Endocrinol Branch, NIH, Bethesda, MD 20892 USA. NHLBI, Off Biostat Res, Div Epidemiol & Clin Applicat, NIH, Bethesda, MD 20892 USA. Mayo Clin & Mayo Fdn, Dept Lab Med & Pathol, Rochester, MN 55905 USA. RP Stratakis, CA (reprint author), NICHHD, Unit Genet & Endocrinol, Dev Endocrinol Branch, NIH, Bldg 10,Room 10N262,10 Ctr Dr,MSC1862, Bethesda, MD 20892 USA. NR 44 TC 264 Z9 270 U1 0 U2 1 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0964-6906 J9 HUM MOL GENET JI Hum. Mol. Genet. PD DEC 12 PY 2000 VL 9 IS 20 BP 3037 EP 3046 DI 10.1093/hmg/9.20.3037 PG 10 WC Biochemistry & Molecular Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Genetics & Heredity GA 384UY UT WOS:000165965400013 PM 11115848 ER PT J AU Marler, JR Tilley, BC Lu, M Brott, TG Lyden, PC Grotta, JC Broderick, JP Levine, SR Frankel, MP Horowitz, SH Haley, EC Lewandowski, CA Kwiatkowski, TP AF Marler, JR Tilley, BC Lu, M Brott, TG Lyden, PC Grotta, JC Broderick, JP Levine, SR Frankel, MP Horowitz, SH Haley, EC Lewandowski, CA Kwiatkowski, TP CA NINDS rt-PA Stroke Study Grp TI Early stroke treatment associated with better outcome - The NINDS rt-PA Stroke Study SO NEUROLOGY LA English DT Article ID TISSUE PLASMINOGEN-ACTIVATOR; URGENT THERAPY; EMBOLIC STROKE; REDUCTION; MINUTES; DAMAGE AB Background: The National Institute of Neurological Disorders and Stroke (NINDS) rt-PA Stroke Study showed a similar percentage of intracranial hemorrhage and good outcome in patients 3 months after stroke treatment given 0 to 90 minutes and 91 to 180 minutes after stroke onset. At 24 hours after stroke onset more patients treated 0 to 90 compared to 91 to 180 minutes after stroke onset had improved by four or more points on the NIH Stroke Scale (NIHSS). The authors performed further analyses to characterize the relationship of onset-to-treatment time (OTT) to outcome at 3 months, early improvement at 24 hours, and intracranial hemorrhage within 36 hours. Methods: Univariate analyses identified potentially confounding variables associated with OTT that could mask an OTT-treatment interaction. Tests for OTT-treatment interactions adjusting for potential masking confounders were performed. An OTT-treatment interaction was considered significant if p less than or equal to 0.10, implying that treatment effectiveness was related to OTT. Results: For 24-hour improvement, there were no masking confounders identified and there was an OTT-treatment interaction (p = 0.08). For 3-month favorable outcome, the NIHSS met criteria for a masking confounder. After adjusting for NIHSS as a covariate, an OTT-treatment interaction was detected (p = 0.09): the adjusted OR (95% CI) for a favorable 3-month outcome associated with recombinant tissue-type plasminogen activator (rt-PA) was 2.11 (1.33 to 3.35) in the 0 to 90 minute stratum and 1.69 (1.09 to 2.62) in the 91 to 180 minute stratum. In the group treated with rt-PA, after adjusting for baseline NIHSS, an effect of OTT on the occurrence of intracranial hemorrhage was not detected. Conclusions: If the NINDS rt-PA Stroke Trial treatment protocol is followed, this analysis suggests that patients treated 0 to 90 minutes from stroke onset with rt-PA have an increased odds of improvement at 24 hours and favorable S-month outcome compared to patients treated later than 90 minutes. No effect of OTT on intracranial hemorrhage was detected within the group treated with rt-PA, possibly due to low power. C1 NINDS, Ctr Neurosci, Bethesda, MD 20892 USA. RP Marler, JR (reprint author), NINDS, Ctr Neurosci, Room 2216,6001 Execut Blvd, Bethesda, MD 20892 USA. FU NINDS NIH HHS [N01-NS-02374, N01-NS-02377, N01-NS-02382] NR 19 TC 476 Z9 491 U1 0 U2 10 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0028-3878 J9 NEUROLOGY JI Neurology PD DEC 12 PY 2000 VL 55 IS 11 BP 1649 EP 1655 PG 7 WC Clinical Neurology SC Neurosciences & Neurology GA 381NY UT WOS:000165770600010 PM 11113218 ER PT J AU Gran, B Tranquill, LR Chen, M Bielekova, B Zhou, W Dhib-Jalbut, S Martin, R AF Gran, B Tranquill, LR Chen, M Bielekova, B Zhou, W Dhib-Jalbut, S Martin, R TI Mechanisms of immunomodulation by glatiramer acetate SO NEUROLOGY LA English DT Article ID MYELIN BASIC-PROTEIN; T-CELL-RECEPTOR; EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS; ANTIGEN-PRESENTING CELLS; EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS; REMITTING MULTIPLE-SCLEROSIS; SYNTHETIC COPOLYMER-1; BYSTANDER SUPPRESSION; LARGE NUMBERS; RELAPSE RATE AB Objective: To define the mechanism of action of glatiramer acetate (GA; formerly known as copolymer-1) as an immunomodulatory treatment for MS. Background: The proposed mechanisms of action of GA include 1) functional inhibition of myelin-reactive T cells by human leukocyte antigen (HLA) blocking, 2) T-cell receptor (TCR) antagonism, and 3) induction of T helper 2 (Th2) immunomodulatory cells. In this report, the authors examined the effects of GA on the functional activation of human T-cell clones (TCC) specific for myelin basic protein (MBP) and for foreign antigens. Several questions were addressed: Is the inhibitory effect of GA specific for autoantigens? Is it mediated by blocking the interaction between peptide and HLA molecule? Is GA a partial agonist or TCR antagonist, or does it induce anergy? Does it induce Th2 modulatory T cells? Methods: The effects of GA on antigen-induced activation of human TCC specific for MBP, influenza virus hemagglutinin, and Borrelia burgdorferi were studied by proliferation and cytokine measurements, TCR downmodulation, and anergy assays. GA-specific TCC were generated in vitro from the peripheral blood of patients and healthy controls by limiting dilution. Results: GA more strongly inhibited the proliferation of MBP, as compared with foreign antigen-specific TCC; in some MBP-specific TCC, the production of Th1-type cytokines was preferentially inhibited. In addition to HLA competition, the induction of anergy, but not direct TCR antagonism, was observed. Numerous GA-specific TCC were generated from the peripheral blood of both MS patients and normal controls, and a fraction of these showed a Th2 phenotype. Conclusions: This study confirms a preferential inhibitory effect of GA on autoreactive TCC. With respect to cellular mechanisms, although HLA competition appears to play the most important role in functional inhibition in vitro, a direct effect on the TCR may be involved at least in some autoreactive T cells as shown by anergy induction. Although not confirmed at the clonal level, it is demonstrated further that GA induces T cells that crossreact with myelin proteins. GA-specific, Th2-modulatory cells may play an important role in mediating the effect of the drug in vivo. C1 NINDS, Cellular Immunol Sect, Neuroimmunol Branch, NIH, Bethesda, MD 20892 USA. Univ Maryland, Sch Med, Dept Neurol, Baltimore, MD 21201 USA. RP Martin, R (reprint author), NINDS, Cellular Immunol Sect, Neuroimmunol Branch, NIH, Bldg 10,Room 5B-16,10 Ctr Dr MSC 1400, Bethesda, MD 20892 USA. RI Gran, Bruno/A-2288-2013 OI Gran, Bruno/0000-0001-6384-2342 FU NINDS NIH HHS [K24 NS02082-01] NR 39 TC 127 Z9 133 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0028-3878 J9 NEUROLOGY JI Neurology PD DEC 12 PY 2000 VL 55 IS 11 BP 1704 EP 1714 PG 11 WC Clinical Neurology SC Neurosciences & Neurology GA 381NY UT WOS:000165770600018 PM 11113226 ER PT J AU Baker, B Paquette, M Szalai, JP Driver, H Perger, T Helmers, K O'Kelley, B Tobe, S AF Baker, B Paquette, M Szalai, JP Driver, H Perger, T Helmers, K O'Kelley, B Tobe, S TI The influence of marital adjustment on 3-year left ventricular mass and ambulatory blood pressure in mild hypertension SO ARCHIVES OF INTERNAL MEDICINE LA English DT Article ID INTERACTION CODING SYSTEM; JOB STRAIN; EMPIRICAL-EVALUATION; RISK-FACTORS; SCALE; CONFLICT; ASSOCIATION; DISTRESS; DISEASE AB Background: Of psychosocial stressors, job strain has been associated with a sustained increase in blood pressure. The impact of marital factors on blood pressure and target organ has not been explored. Objectives: To evaluate whether marital adjustment, measured at baseline by self-report (Dyadic Adjustment Scale) influences left ventricular mass index (LVMI) and ambulatory blood pressure measured over 3 years in patients with mild hypertension. Methods: A prospective cohort study was conducted on 103 cohabiting males or females, including 72 with technically adequate echocardiograms, who at baseline were unmedicated, employed, and living with a significant other, all for a minimum of 6 months and had repeated elevated office diastolic blood pressure. Main Outcome Measures: Left ventricular mass by M-mode echocardiography indexed to body surface area and blood pressure were measured by ambulatory blood pressure every 15 minutes (daytime) and hourly between 11 PM and 7 AM. Results: Marital adjustment, smoking, drinking, and baseline LVMI contributed significantly to the prediction of 3-year LVMI (semipartial correlation, sr(2), 0.04, 0.07, 0.03, and 0.22, P = .03, .008, .08, and <.001, respectively) together accounting for 36% of the total variability in follow-up LVMI. Three-year ambulatory blood pressure measures were not significantly related to marital adjustment but there were correlations with Dyadic Adjustment Scale subscales. Low or high levels of spousal contact during 3-year ambulatory blood pressure monitoring were associated with an increase or decrease of 3-year, 24-hour diastolic blood pressure, consistent with the quality of marital adjustment (P = .04) or marital satisfaction (Dyadic Adjustment Scale subscale, P = .008). Conclusions: In a cohort of subjects with mild essential hypertension, marital adjustment had an influence on 3-year LVMI. Depending on the quality of marital adjustment, spousal contact at 3 years was associated with an increase or decrease of 3-year diastolic blood pressure. Confirmation of these results, including objective marital assessment and the participation of normotensive subjects, is required. C1 Univ Toronto, Toronto, ON, Canada. Univ Hlth Network, Dept Psychiat, Toronto, ON, Canada. Univ Hlth Network, Div Cardiol, Toronto, ON, Canada. St Michaels Hosp, Div Cardiol, Toronto, ON M5B 1W8, Canada. Sunnybrook & Womens Hlth Sci Ctr, Dept Res Design & Biostat, Toronto, ON, Canada. Sunnybrook & Womens Hlth Sci Ctr, Div Nephrol, Toronto, ON, Canada. NINR, NIH, Bethesda, MD 20892 USA. RP Baker, B (reprint author), Toronto Western Hosp, 3D Edith Cavell Wing,399 Bathurst St, Toronto, ON M5T 2S8, Canada. NR 46 TC 40 Z9 40 U1 1 U2 2 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0003-9926 J9 ARCH INTERN MED JI Arch. Intern. Med. PD DEC 11 PY 2000 VL 160 IS 22 BP 3453 EP 3458 DI 10.1001/archinte.160.22.3453 PG 6 WC Medicine, General & Internal SC General & Internal Medicine GA 380RX UT WOS:000165718200014 PM 11112239 ER PT J AU Zhou, JZ Zhang, F Zhang, YX AF Zhou, JZ Zhang, F Zhang, YX TI Corticosterone inhibits generation of long-term potentiation in rat hippocampal slice: involvement of brain-derived neurotrophic factor SO BRAIN RESEARCH LA English DT Article DE corticosterone; long-term potentiation; paired-pulse facilitation; synaptic plasticity; synaptic transmission; reverse-transcription-polymerase chain reaction; neurotrophin; nerve growth factor (NGF); brain-derived neurotrophic factor (BDNF); neurotrophic factor-3 (NT-3) ID INDUCED EXTRACELLULAR ACCUMULATION; DEVELOPING NEUROMUSCULAR SYNAPSES; PRIMED BURST POTENTIATION; SYNAPTIC TRANSMISSION; ALZHEIMERS-DISEASE; MESSENGER-RNAS; GLUCOCORTICOIDS; STRESS; PLASTICITY; BDNF AB In the present study, the effect of corticosterone (CORT) on the generation of long-term potentiation (LTP) and its underlying mechanism involving neurotrophin gene expression in CA1 synopses of mt hippocampal slice were examined. Our experimental results showed incubation of hippocampal slice with CORT for 3 h had no effect on either the slope or amplitude of excitatory postsynaptic potentials (EPSP) evoked in hippocampal CA1 pyramidal dentrites, indicating no marked change in basal synaptic transmission. However, when tetanic stimulation (100 pulses, 100 Hz) was delivered to the Schaffer collateral pathway, CORT application significantly attenuated the tetanus-induced increases of both EPSP slope and amplitude, demonstrating an inhibitory effect of CORT on LTP generation. In addition. CORT treatment significantly reduced both slope and amplitude ratios of the second evoked EPSP to the first one when paired-pulse facilitation (PPF) was established at different interpulse intervals from 20 to 40 ms, suggesting that a presynaptic mechanism may be involved in CORT-induced hippocampal synaptic plasticity. Reverse-transcription polymerase chain reaction (RT-PCR) analysis showed that CORT-treated hippocampal CA1 cells underwent a significant decrease in the expression of mRNA for nerve growth factor-beta (NGF-beta) and brain-derived neurotrophic factor (BDNF), but not for neurotrophin-3 (NT-3) compared with those in control. Moreover, BDNF co-applied with CORT significantly antagonized CORT-induced deficit in PPF. Taken together, the present results suggest that CORT-induced inhibition of LTP may be, at least to some extent, mediated by a presynaptic mechanism and decrease in the BDNF expression in rat hippocampal CA1 cells induced by CORT may partially account for this presynaptic mechanism. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Beijing Inst Pharmacol & Toxicol, Neuropharmacol Lab, Beijing 100850, Peoples R China. RP NICHD, Dev Neurobiol Lab, Unit Synapt Dev & Plast, NIH, Bldg 49 Rm 6A67, Bethesda, MD 20892 USA. EM jzhou@codon.nih.gov NR 46 TC 44 Z9 48 U1 2 U2 6 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0006-8993 EI 1872-6240 J9 BRAIN RES JI Brain Res. PD DEC 8 PY 2000 VL 885 IS 2 BP 182 EP 191 DI 10.1016/S0006-8993(00)02934-6 PG 10 WC Neurosciences SC Neurosciences & Neurology GA 385VX UT WOS:000166026800005 PM 11102572 ER PT J AU Chesley, A Lundberg, MS Asai, T Xiao, RP Ohtani, S Lakatta, EG Crow, MT AF Chesley, A Lundberg, MS Asai, T Xiao, RP Ohtani, S Lakatta, EG Crow, MT TI The beta(2)-adrenergic receptor delivers an antiapoptotic signal to cardiac myocytes through G(i)-dependent coupling to phosphatidylinositol 3 '-kinase SO CIRCULATION RESEARCH LA English DT Article DE apoptosis; beta-adrenergic receptors; cardiomyocytes; hypoxia; phosphatidylinositol; 3 '-kinase; G(i) proteins ID RAT VENTRICULAR MYOCYTES; ACTIVATED PROTEIN-KINASE; INDUCED APOPTOSIS; GROWTH-FACTOR; PATHWAY; CELL; HYPERTROPHY; DOWNSTREAM; INHIBITION; SURVIVAL AB Recent studies have shown that chronic beta -adrenergic receptor (beta -AR) stimulation alters cardiac myocyte survival in a receptor subtype-specific manner. We examined the effect of selective beta (1)- and beta (2)-AR subtype stimulation on apoptosis induced by hypoxia or H2O2 in rat neonatal cardiac myocytes. Although neither beta (1)- nor beta (2)-AR stimulation had any significant effect on the basal level of apoptosis, selective beta (2)-AR stimulation protected myocytes from apoptosis. beta (2)-AR stimulation markedly increased mitogen-activated protein kinase/extracellular signal-regulated protein kinase (MAPK/ERK) activation as well as phosphatidylinositol-3'-kinase (PI-3K) activity and Akt/protein kinase B phosphorylation. beta (1)-AR stimulation also markedly increased MAPK/ERK activation but only minimally activated PI-3K and Akt, Pretreatment with pertussis toxin blocked beta (2)-AR-mediated protection from apoptosis as well as the beta (2)-AR-stimulated changes in MAPK/ERK, PI-3K, and Akt/protein kinase B. The selective PI-3K inhibitor, LY 294002, also blocked beta (2)-AR-mediated protection, whereas inhibition of MAPK/ERK activation at an inhibitor concentration that blocked agonist-induced activation but not the basal level of activation had no effect on beta (2)-AR-mediated protection. These findings demonstrate that beta (2)-ARs activate a PI-3K-dependent, pertussis toxin-sensitive signaling pathway in cardiac myocytes that is required for protection from apoptosis-inducing stimuli often associated with ischemic stress. C1 NIA, Gerontol Res Ctr, Cardiovasc Sci Lab, NIH, Baltimore, MD 21224 USA. RP Crow, MT (reprint author), NIA, Gerontol Res Ctr, Cardiovasc Sci Lab, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 25 TC 242 Z9 255 U1 1 U2 7 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7330 J9 CIRC RES JI Circ.Res. PD DEC 8 PY 2000 VL 87 IS 12 BP 1172 EP 1179 PG 8 WC Cardiac & Cardiovascular Systems; Hematology; Peripheral Vascular Disease SC Cardiovascular System & Cardiology; Hematology GA 384EX UT WOS:000165930800017 PM 11110775 ER PT J AU Moody, TW Jensen, RT Garcia, L Leyton, J AF Moody, TW Jensen, RT Garcia, L Leyton, J TI Nonpeptide neuromedin B receptor antagonists inhibit the proliferation of C6 cells SO EUROPEAN JOURNAL OF PHARMACOLOGY LA English DT Article DE neuromedin B receptor antagonist, nonpeptide; Ca2+, cytosolic; c-fos mRNA; focal adhesion kinase; proliferation ID GASTRIN-RELEASING PEPTIDE; LUNG-CANCER CELLS; SWISS 3T3 CELLS; BOMBESIN-LIKE PEPTIDES; HIGH-AFFINITY; CYTOSOLIC CALCIUM; GROWTH-FACTORS; RAT; CLONING; LINES AB The ability of nonpeptide antagonists to interact with neuromedin B receptors on C6 cells was investigated. 2-[3-(2,6-Diisopropylphenyl)-ureido]3-(1H-indol-3-yl)-2-methyl-N-(1-pyridin-2-yl-cyclohexylmethyl)-proprionate (PD165929), 3-(1H-indol-3-yl)-2-methyl-2-[3(4-nitro-phenyl)-ureido]-N-(1-pyridin-2-yl-cyclohexylmethyl)-propionamide (PD168368) and 3-(1H-indol-3-yl)-N-[1-(5-methoxy-pyridin-2-yl)-cyclohexylmethyl]-2-methyl-2-[3-(4-mitro-phenyl)-ureido]-propionamide (PD176252) inhibited (I-125-Tyr(0))neuromedin B binding with IC50 values of 2000, 40 and 50 nM, respectively. Because neuromedin B is a G-protein coupled serpentine receptor, the effects of neuromedin B antagonists on second messenger production and proliferation were investigated. PD168368 inhibited the ability of 10 nM neuromedin B to cause elevation of cytosolic Ca2+, whereas it had no effect on basal cytosolic Ca2+. PD168368 inhibited the ability of 100 nM neuromedin B to cause elevation of c-fos mRNA. Also, PD168368 in a dose-dependent manner inhibited the ability of 100 nM neuromedin B to cause phosphorylation of focal adhesion kinase. Using a [3-(4,5 dimethylthiazol-2-yl)-2.5-diphenyl-2H-tetrazolium bromide] assay, the order of antagonist potency to inhibit C6 proliferation was PD168368 = PD176252 > PD165929. Also, 1 muM PD168368 and PD176252 significantly inhibited colony number using a proliferation assay in vitro. PD168368 significantly inhibited C6 xenograft growth in nude mice in vivo. These results indicate that PD168368 is a C6 cell neuromedin B receptor antagonist, which inhibits proliferation. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NCI, Cell & Canc Biol Dept, Med Branch, Rockville, MD 20850 USA. NIH, Digest Dis Branch, Bethesda, MD 20892 USA. RP Moody, TW (reprint author), NCI, Cell & Canc Biol Dept, Med Branch, Bldg KWC,Rm 300,9610 Med Ctr Dr, Rockville, MD 20850 USA. RI Garcia-Marin, Luis /L-4680-2014 OI Garcia-Marin, Luis /0000-0002-1795-7381 NR 43 TC 18 Z9 18 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0014-2999 J9 EUR J PHARMACOL JI Eur. J. Pharmacol. PD DEC 8 PY 2000 VL 409 IS 2 BP 133 EP 142 DI 10.1016/S0014-2999(00)00828-1 PG 10 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 381QG UT WOS:000165774000005 PM 11104826 ER PT J AU Brasaemle, DL Rubin, B Harten, IA Gruia-Gray, J Kimmel, AR Londos, C AF Brasaemle, DL Rubin, B Harten, IA Gruia-Gray, J Kimmel, AR Londos, C TI Perilipin A increases triacylglycerol storage by decreasing the rate of triacylglycerol hydrolysis SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HORMONE-SENSITIVE LIPASE; CHOLESTERYL ESTER HYDROLASE; DEPENDENT PROTEIN-KINASE; ACYL-COA SYNTHETASE; LEYDIG TUMOR-CELLS; SACCHAROMYCES-CEREVISIAE; LIPID PARTICLES; ENDOPLASMIC-RETICULUM; MEMBRANE-FRACTION; MAMMALIAN-CELLS AB The perilipins are the most abundant proteins at the surfaces of lipid droplets in adipocytes and are also found in steroidogenic cells. To investigate perilipin function, perilipin A, the predominant isoform, was ectopically expressed in fibroblastic 3T3-L1 pre-adipocytes that normally lack the perilipins. In control cells, fluorescent staining of neutral lipids with Bodipy 493/503 showed a few minute and widely dispersed lipid droplets, while in cells stably expressing perilipin A, the lipid droplets were more numerous and tightly clustered in one or two regions of the cytoplasm. Immunofluorescence microscopy revealed that the ectopic perilipin A localized to the surfaces of the tiny clustered lipid droplets; subcellular fractionation of the cells using sucrose gradients confirmed that the perilipin A localized exclusively to lipid droplets. Cells expressing perilipin A stored 6-30-fold more triacylglycerol than control cells due to reduced lipolysis of triacylglycerol stores. The lipolysis of stored triacylglycerol was 5 times slower in lipid-loaded cells expressing perilipin A than in lipid-loaded control cells, when triacylglycerol synthesis was blocked with 6 muM triacsin C. This stabilization of triacylglycerol was not due to the suppression of triacylglycerol lipase activity by the expression of perilipin A. We conclude that perilipin A increases the triacylglycerol content of cells by forming a barrier that reduces the access of soluble lipases to stored lipids, thus inhibiting triacylglycerol hydrolysis. These studies suggest that perilipin A plays a major role in the regulation of triacylglycerol storage and lipolysis in adipocytes. C1 Rutgers State Univ, Dept Nutrit Sci, New Brunswick, NJ 08901 USA. Med Coll Penn & Hahnemann Univ, Dept Biochem, Philadelphia, PA 19129 USA. NIDDK, Mol Mechanisms Dev Sect, NIH, Bethesda, MD 20892 USA. NIDDK, Membrane Regulat Sect, Cellular & Dev Biol Lab, NIH, Bethesda, MD 20892 USA. RP Brasaemle, DL (reprint author), Rutgers State Univ, Dept Nutrit Sci, 96 Lipman Dr, New Brunswick, NJ 08901 USA. OI Brasaemle, Dawn/0000-0002-8553-8285 NR 50 TC 296 Z9 314 U1 3 U2 20 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 8 PY 2000 VL 275 IS 49 BP 38486 EP 38493 DI 10.1074/jbc.M007322200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 381AH UT WOS:000165739800048 PM 10948207 ER PT J AU Longenecker, KL Zhang, BL Derewenda, U Sheffield, PJ Dauter, Z Parsons, JT Zheng, Y Derewenda, ZS AF Longenecker, KL Zhang, BL Derewenda, U Sheffield, PJ Dauter, Z Parsons, JT Zheng, Y Derewenda, ZS TI Structure of the BH domain from Graf and its implications for Rho GTPase recognition SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID FOCAL ADHESION KINASE; ACTIVATING-PROTEIN; CRYSTAL-STRUCTURE; P50RHOGAP; COMPLEX; BINDING; P190; P85 AB Cellular signaling by small G-proteins is down-regulated by GTPase-activating proteins (GAPs), which increase the rate of GTP hydrolysis, The GTPase regulator associated with focal adhesion kinase (Graf) exhibits GAP activity toward the RhoA and Cdc42 GTPases, but is only weakly active toward the closely related Rad. We determined the crystal structure of a 231-residue fragment of Graf (GrafGAP), a domain containing the GAP activity, at 2.4-Angstrom resolution The structure clarifies the boundaries of the functional domain and yields insight to the mechanism of substrate recognition. Modeling its interaction with substrate suggested that a favorable interaction with Glu-95 of Cdc42 (Glu-97 of RhoA) would be absent with the corresponding Ala-95 of Rad. Indeed, GrafGAP activity is diminished similar to 40-fold toward a Cdc42 E95A mutant, whereas a similar to 10-fold increase is observed for a Rad A95E mutant. The GrafGAP epitope that apparently interacts with Glu-95(Glu-97) contains Asn-225, which was recently found mutated in some myeloid leukemia patients. We conclude that position 95 of the GTPase is an important determinant for GrafGAP specificity in cellular function and tumor suppression. C1 Univ Virginia Hlth Syst, Dept Mol Physiol & Biol Phys, Charlottesville, VA 22908 USA. Univ Virginia Hlth Syst, Dept Microbiol, Charlottesville, VA 22908 USA. Univ Tennessee, Dept Biochem, Memphis, TN 38163 USA. NCI, Frederick, MD 21702 USA. Brookhaven Natl Lab, Upton, NY 11973 USA. Univ Virginia, Ctr Canc, Charlottesville, VA 22908 USA. RP Derewenda, ZS (reprint author), Univ Virginia Hlth Syst, Dept Mol Physiol & Biol Phys, Charlottesville, VA 22908 USA. EM ZSD4N@virginia.edu RI Zheng, Yi/J-7235-2015 OI Zheng, Yi/0000-0001-7089-6074 FU NHLBI NIH HHS [HL48807]; NIGMS NIH HHS [GM60523] NR 27 TC 16 Z9 17 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 8 PY 2000 VL 275 IS 49 BP 38605 EP 38610 DI 10.1074/jbc.M007574200 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 381AH UT WOS:000165739800063 PM 10982819 ER PT J AU Shakur, Y Takeda, K Kenan, Y Yu, ZX Rena, G Brandt, D Houslay, MD Degerman, E Ferrans, VJ Manganiello, VC AF Shakur, Y Takeda, K Kenan, Y Yu, ZX Rena, G Brandt, D Houslay, MD Degerman, E Ferrans, VJ Manganiello, VC TI Membrane localization of cyclic nucleotide phosphodiesterase 3 (PDES) - Two N-terminal, domains are required for the efficient targeting to, and association of, PDE3 with endoplasmic reticulum SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID INHIBITED CAMP-PHOSPHODIESTERASE; AMP-SPECIFIC PHOSPHODIESTERASE; MICROSOMAL ALDEHYDE DEHYDROGENASE; GREEN FLUORESCENT PROTEIN; SARCOPLASMIC-RETICULUM; PLASMA-MEMBRANE; RAT ADIPOCYTES; PHOSPHATIDYLINOSITOL 3-KINASE; SUBCELLULAR-LOCALIZATION; SELECTIVE INHIBITOR AB Subcellular localization of cyclic nucleotide phosphodiesterases (PDEs) may be important in compartmentalization of cAMP/cGMP signaling responses. In 3T3-L1 adipocytes, mouse (M) PDE3B was associated with the endoplasmic reticulum (ER) as indicated by its immunofluorescent colocalization with the ER protein BiP and subcellular fractionation studies. In transfected NIH 3006 or COS-7 cells, recombinant wild-type PDE3A and PDE3B isoforms were both found almost exclusively in the ER. The N-terminal portion of PDE3 can be arbitrarily divided into region 1 (aa 1-300), which contains a large hydrophobic domain with six predicted transmembrane helices, followed by region 2 (aa 301-500) containing a smaller hydrophobic domain (of similar to 50 aa). To investigate the role of regions 1 and 2 in membrane association, we examined the subcellular localization of a series of catalytically active, Flag-tagged N-terminal-truncated human (H) PDE3A and MPDE3B recombinants, as well as a series of fragments from regions 1 and 2 of MPDESB synthesized as enhanced green fluorescent (EGFP) fusion proteins in COS-7 cells. In COS-7 cells, the localization of a mutant HPDE3A, lacking the first 189 amino acids (aa) and therefore four of the six predicted transmembrane helices (H3A-Delta 189), was virtually identical to that of the wild type. R3B-Delta 302 (lacking region 1) and H3A-Delta 397 (lacking region 1 as well as part of region 2) retained, to different degrees, the ability to associate with membranes, albeit less efficiently than H3A-Delta 189. Proteins that lacked both regions I and 2, H3A-Delta 510 and M3B-Delta 604, did not associate with membranes. Consistent with these findings, region I EGFP-MPDE3B fusion proteins colocalized with the ER, whereas region 2 EGFP fusion proteins were diffusely distributed. Thus, some portion of the N-terminal hydrophobic domain in region 1 plus a second domain in region 2 are important for efficient membrane association/targeting of PDES. C1 NHLBI, Pulm Crit Care Med Branch, NIH, Bethesda, MD 20892 USA. NHLBI, Pathol Sect, NIH, Bethesda, MD 20892 USA. Otsuka Amer Pharmaceut Inc, Rockville, MD 20850 USA. Univ Glasgow, Mol Pharmacol Grp, Div Biochem & Mol Biol, Glasgow G12 8Q0, Lanark, Scotland. Univ Lund, Dept Cell & Mol Biol, Sect Mol Signaling, S-22100 Lund, Sweden. RP Manganiello, VC (reprint author), NHLBI, Pulm Crit Care Med Branch, NIH, Bldg 10,Rm 5N307,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Houslay, Miles/A-6825-2011 NR 70 TC 81 Z9 84 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 8 PY 2000 VL 275 IS 49 BP 38749 EP 38761 DI 10.1074/jbc.M001734200 PG 13 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 381AH UT WOS:000165739800084 PM 10952971 ER PT J AU Yang, Y Hwang, CK Junn, E Lee, G Mouradian, MM AF Yang, Y Hwang, CK Junn, E Lee, G Mouradian, MM TI ZIC2 and Sp3 repress Sp1-induced activation of the human D-1A dopamine receptor gene SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID TRANSCRIPTION FACTORS; BINDING-SITES; CELL LINEAGE; SP FAMILY; EXPRESSION; PROMOTER; ENHANCEMENT; AMPHETAMINE; MODULATION; AGONISTS AB The human D-1A dopamine receptor is transcribed from a tissue-specific regulated gene under the control of two promoters. An activator region (AR1) located between nucleotides -1154 and -1136 (relative to the first ATG) enhances transcription from the upstream promoter that is active in the brain. In this investigation, we sought to identify the nuclear factors that regulate the D-1A gene through their binding to AR1 using yeast one-hybrid screening. Sp3 and Zic2 were among the positive clones isolated. Although Sp1 was not isolated from this screening and purified Sp1 alone does not bind to AR1 in gel shift experiments, this general transcription factor binds to AR1 in the presence of D-1A expressing NS20Y nuclear extract and activates the D-1A promoter. Thus, Sp1 appears to require an unknown factor(s) or post-translational modification to interact with AR1. On the other hand, Zic2 and Sp3 inhibit Sp1-induced activation of the D-1A gene in an AR1-dependent manner. Zic2 and D-1A genes have reciprocal brain regional distributions; Zic2 is expressed primarily in the cerebellum, and D-1A is highly expressed in corpus striatum. These observations collectively suggest that one of the physiologic functions of Zic2 is repression of D-1A gene transcription and that the intracellular balance among Sp1, Sp3 and Zic2 is important for regulating the tissue-specific expression of this dopamine receptor. C1 NINDS, Genet Pharmacol Unit, Expt Therapeut Branch, NIH, Bethesda, MD 20892 USA. RP Mouradian, MM (reprint author), NINDS, Genet Pharmacol Unit, Expt Therapeut Branch, NIH, 10 Ctr Dr,MSC 1406, Bethesda, MD 20892 USA. OI Mouradian, M. Maral/0000-0002-9937-412X NR 43 TC 43 Z9 45 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 8 PY 2000 VL 275 IS 49 BP 38863 EP 38869 DI 10.1074/jbc.M007906200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 381AH UT WOS:000165739800098 PM 10984499 ER PT J AU Bommakanti, RK Vinayak, S Simonds, WF AF Bommakanti, RK Vinayak, S Simonds, WF TI Dual regulation of Akt/protein kinase B by heterotrimeric G protein subunits SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article; Proceedings Paper CT American-Society-of-Biochemistry-and-Molecular-Biology Annual Meeting CY MAY 15-20, 1998 CL WASHINGTON, D.C. SP Amer Soc Biochem & Molec Biol ID BETA-GAMMA-SUBUNITS; ACTIVATED PHOSPHATIDYLINOSITOL 3-KINASE; GLYCOGEN-SYNTHASE KINASE-3; PLECKSTRIN HOMOLOGY DOMAIN; FACTOR-I RECEPTOR; COUPLED RECEPTORS; PHOSPHOINOSITIDE 3-KINASE-GAMMA; MOLECULAR-CLONING; AKT PROTOONCOGENE; INSULIN AB While positive regulation of c-Akt (also known as protein kinase B) by receptor tyrosine kinases is well documented, compounds acting through G protein-coupled receptors can also activate Akt and its downstream targets. We therefore explored the role of G protein subunits in the regulation of Akt in cultured mammalian cells,In HEK-293 and COS-7 cells transiently transfected with beta (2)-adrenergic or m2 muscarinic receptors, respectively, treatment with agonist-induced phosphorylation of Akt at serine 473 as evidenced by phosphoserine-specific immunoblots. This effect was blocked by the phosphatidylinositol-3-OH kinase inhibitor LY294002 and wild-type G alpha (il), and was not duplicated by co-transfection of the constitutively active G alpha (s)-Q227L or G alpha (i)-Q204L mutant, Co-transfection of G beta (1), G beta (2) but not G beta (5) together with G gamma (2) activated the kinase when assayed in vitro following immunoprecipitation of the epitope-tagged enzyme. In contrast, constitutively activated G protein subunits representing the four G alpha subfamilies were found unable to activate Akt in either cell line. The latter results are in disagreement with a report by Murga et al. (Murga, C., Laguinge, L., Wetzker, R., Cuadrado, A., and Gutkind, J. S. (1998) J, Biol. Chem. 273, 19080-19085) that described activation of Akt in response to mutationally activated G alpha (q) and G alpha (i) transfection in COS cells. To the contrary, in our experiments G alpha (q)-Q209L inhibited Akt activation resulting from beta gamma or mutationally activated H-Ras co-transfection in these cells. In HEK-293 cells G alpha (q)-Q209L transfection inhibited insulin-like growth factor-1 activation of epitope-tagged Akt. In m1 muscarinic receptor transfected HEK-293 cells, carbachol inhibited insulin-like growth factor-1 stimulated phosphorylation at Ser(473) of endogenous Akt in an atropine-reversible fashion. me conclude that G proteins can regulate Akt by two distinct and potentially opposing mechanisms: activation by G beta gamma heterodimers in a phosphatidylinositol-3-OH kinase-dependent fashion, and inhibition mediated by G alpha (q). This work identifies Akt as a novel point of convergence between disparate signaling pathways. C1 NIDDK, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. RP Simonds, WF (reprint author), NIDDK, Metab Dis Branch, NIH, Bldg 10,Rm 8C-101,10 Ctr Dr,MSC 1752, Bethesda, MD 20892 USA. NR 59 TC 80 Z9 80 U1 0 U2 3 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 8 PY 2000 VL 275 IS 49 BP 38870 EP 38876 DI 10.1074/jbc.M007403200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 381AH UT WOS:000165739800099 PM 10986289 ER PT J AU Witta, J Baffi, JS Palkovits, M Mezey, E Castillo, SO Nikodem, VM AF Witta, J Baffi, JS Palkovits, M Mezey, E Castillo, SO Nikodem, VM TI Nigrostriatal innervation is preserved in Nurr1-null mice, although dopaminergic neuron precursors are arrested from terminal differentiation SO MOLECULAR BRAIN RESEARCH LA English DT Article DE nurr1; dopamine; cholecystokinin; DiI; substantia nigra; ventral tegmental area ID NURR1-DEFICIENT MICE; NGFI-B; TYROSINE-HYDROXYLASE; CHOLECYSTOKININ-LIKE; NUCLEAR RECEPTOR; SONIC HEDGEHOG; MESSENGER-RNAS; ADULT-RAT; EXPRESSION; BRAIN AB Various factors, including the orphan nuclear receptor Nurr1, have been implicated in dopamine biosynthesis, but many of the specific events involved in this process have to be determined. Using genetic manipulations in mice, the obligatory role for Nurr1 in dopamine (DA) biosynthesis has been documented; however, the mechanism remains unclear. DA biosynthetic enzymes, transporters and receptors are absent in the substantia nigra (SN) and the ventral tegmental area (VTA) of Nurr1-null neonates. The current study establishes that the loss of Nurr1 function does not affect the normal ventralization of neuroepithelial cells to the ventral midbrain, their differentiation into neurons, and their topographical pattern in the SN and VTA, Furthermore, the absence of Nurr1 does not affect the survival of these DA precursor cells in the ventral midbrain, as determined hv quantative analysis of cells expressing the general neuronal nuclear marker (NeuN) and the TUNEL assay for apoptosis, These neurons express cholecystokinin (CCK), a co-transmitter of dopaminergic neurons in this area. The untranslated exon 1-2 of the Nurr1 gene, which remains intact after homologous recombination, revealed the presence of dopaminergic precursors in the ventral midbrain of the Nurrl-null mice. In addition, these neurons establish their nigrostriatal projections, as shown by axonal transport of a fluorescent tracer, DiI. These results provide evidence that Nurrl is essential for terminal differentiation of the dopaminergic neurons in the ventral midbrain but does not affect the early steps of their neurogenesis, migration, survival and striatal projections. Our findings suggest that activation of Nurr1 might be therapeutically useful in Parkinson's disease. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIDDKD, NIH, Bethesda, MD 20892 USA. NINDS, NIH, Bethesda, MD 20892 USA. NIMH, NIH, Bethesda, MD 20892 USA. RP Nikodem, VM (reprint author), NIDDKD, NIH, Bldg 10,Room 8N317,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Palkovits, Miklos/F-2707-2013; OI Palkovits, Miklos/0000-0003-0578-0387 NR 38 TC 36 Z9 36 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0169-328X J9 MOL BRAIN RES JI Mol. Brain Res. PD DEC 8 PY 2000 VL 84 IS 1-2 BP 67 EP 78 DI 10.1016/S0169-328X(00)00211-4 PG 12 WC Neurosciences SC Neurosciences & Neurology GA 386ZE UT WOS:000166095000008 ER PT J AU Li, CY Xiong, KM Weight, FF AF Li, CY Xiong, KM Weight, FF TI Ethanol inhibition of adenosine 5 '-triphosphate-activated current in freshly isolated adult rat hippocampal CA1 neurons SO NEUROSCIENCE LETTERS LA English DT Article DE adenosine 5 '-triphosphate; ion channels; P2X receptors; hippocampus; alcohol ID EXCITATORY SYNAPTIC TRANSMISSION; MAMMALIAN NEURONS; ION CHANNELS; GUINEA-PIG; ATP; RECEPTORS; RELEASE; SLICES; POTENTIATION; P2X(4) AB The effect of ethanol on current activated by extracellular adenosine 5'-triphosphate (ATP) was studied in freshly isolated adult rat hippocampal CA1 neurons using whole-cell patch-clamp recording. ATP activated an inward current with an EC50 value of 18 muM. The inward current was also activated by 2-methylthio ATP (2-MeSATP) and cr,P-methylene ATP (alpha,beta -MeATP), inhibited by pyridoxal-phosphate-6-azophenyl-2',4'-disulfonic acid (PPADS), and potentiated by Zn2+, Ethanol inhibited current activated by 10 muM ATP with an IC50 value of 83 mM in a voltage-independent manner; Ethanol, 100 mM, shifted the ATP concentration-response curve to the right, increasing the EC50 value for ATP from 18 to 33 muM, but did not reduce the maximal response to ATP. The results suggest that ethanol can inhibit the function of P2X receptors in adult rat hippocampal neurons by decreasing the apparent affinity of the binding site for ATP, (C) 2000 Elsevier Science Ireland Ltd. All rights reserved. C1 NIAAA, Mol & Cellular Neurobiol Lab, NIH, Bethesda, MD 20892 USA. RP Li, CY (reprint author), AstraZeneca CNS Discovery, Dept Neurosci A131, 1800 Concord Pike,POB 15437, Wilmington, DE 19850 USA. NR 27 TC 21 Z9 21 U1 0 U2 0 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0304-3940 J9 NEUROSCI LETT JI Neurosci. Lett. PD DEC 8 PY 2000 VL 295 IS 3 BP 77 EP 80 DI 10.1016/S0304-3940(00)01586-X PG 4 WC Neurosciences SC Neurosciences & Neurology GA 379TL UT WOS:000165658500002 PM 11090978 ER PT J AU Chen, HT Bhandoola, A Difilippantonio, MJ Zhu, J Brown, MJ Tai, XG Rogakou, EP Brotz, TM Bonner, WM Ried, T Nussenzweig, A AF Chen, HT Bhandoola, A Difilippantonio, MJ Zhu, J Brown, MJ Tai, XG Rogakou, EP Brotz, TM Bonner, WM Ried, T Nussenzweig, A TI Response to RAG-mediated V(D)J cleavage by NBS1 and gamma-H2AX SO SCIENCE LA English DT Article ID DOUBLE-STRAND BREAKS; T-CELL; MICE DEFICIENT; RECOMBINATION; DNA; THYMOCYTES; COMPLEX; REPAIR; REARRANGEMENT; EXPRESSION AB Genetic disorders affecting cellular responses to DNA damage are characterized by high rates of translocations involving antigen receptor loci and increased susceptibility to lymphoid malignancies. We report that the Nijmegen breakage syndrome protein (NBS1) and histone gamma -H2AX, which associate with irradiation-induced DNA double-strand breaks (DSBs), are also found at sites of V(D)J (variable, diversity, joining) recombination-induced DSBs. In developing thymocytes, NBS1 and gamma -H2AX form nuclear foci that colocalize with the T cell receptor alpha locus in response to recombination activating gene (RAG) protein-mediated V(D)J cleavage. Our results suggest that surveillance of T cell receptor recombination intermediates by NBS1 and gamma -H2AX may be important for preventing oncogenic translocations. C1 NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. NCI, Dept Genet, NIH, Bethesda, MD 20892 USA. NCI, Mol Pharmacol Lab, NIH, Bethesda, MD 20892 USA. RP Nussenzweig, A (reprint author), NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. RI Brotz, Tilmann/R-6599-2016 OI Brotz, Tilmann/0000-0001-8212-459X FU Intramural NIH HHS [Z99 CA999999] NR 25 TC 238 Z9 246 U1 0 U2 6 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 USA SN 0036-8075 J9 SCIENCE JI Science PD DEC 8 PY 2000 VL 290 IS 5498 BP 1962 EP 1964 DI 10.1126/science.290.5498.1962 PG 3 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 382EL UT WOS:000165810000045 PM 11110662 ER PT J AU Plotkin, SA Aubin, F Robbins, J AF Plotkin, SA Aubin, F Robbins, J TI Letter to the editor SO VACCINE LA English DT Letter ID METAANALYSIS C1 Aventus Pasteur, Doylestown, PA 18901 USA. Lab Fournier, Garches, France. NICHHD, Dev & Mol Immun Lab, NIH, Bethesda, MD 20892 USA. RP Plotkin, SA (reprint author), Aventus Pasteur, 4650 Wismer Rd, Doylestown, PA 18901 USA. NR 7 TC 0 Z9 0 U1 0 U2 0 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 0264-410X J9 VACCINE JI Vaccine PD DEC 8 PY 2000 VL 19 IS 9-10 BP 1003 EP 1003 DI 10.1016/S0264-410X(00)00328-5 PG 1 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 383QW UT WOS:000165894700001 PM 11203497 ER PT J AU Ishikawa, T Maurizi, MR Belnap, D Steven, AC AF Ishikawa, T Maurizi, MR Belnap, D Steven, AC TI ATP-dependent proteases - Docking of components in a bacterial complex SO NATURE LA English DT Article ID PROTEIN C1 NIAMSD, Struct Biol Lab, NIH, Bethesda, MD 20892 USA. NCI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. RP Ishikawa, T (reprint author), NIAMSD, Struct Biol Lab, NIH, Bethesda, MD 20892 USA. NR 11 TC 32 Z9 32 U1 1 U2 2 PU MACMILLAN PUBLISHERS LTD PI LONDON PA PORTERS SOUTH, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD DEC 7 PY 2000 VL 408 IS 6813 BP 667 EP 668 PG 2 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 382GU UT WOS:000165815200034 PM 11130060 ER PT J AU Lekstrom-Himes, JA Gallin, JI AF Lekstrom-Himes, JA Gallin, JI TI Immunodeficiency diseases caused by defects in phagocytes SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Review ID CHRONIC GRANULOMATOUS-DISEASE; CHEDIAK-HIGASHI-SYNDROME; LEUKOCYTE ADHESION DEFICIENCY; BACILLE CALMETTE-GUERIN; BINDING-PROTEIN-EPSILON; SHWACHMAN-DIAMOND-SYNDROME; SEVERE CHRONIC NEUTROPENIA; INTERFERON-GAMMA-RECEPTOR; ACUTE MYELOID-LEUKEMIA; GRANULE DEFICIENCY C1 NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. RP Gallin, JI (reprint author), Bldg 10,Rm 2C146,10 Ctr Dr,MSC 1504, Bethesda, MD 20892 USA. NR 116 TC 203 Z9 211 U1 0 U2 8 PU MASSACHUSETTS MEDICAL SOC PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD DEC 7 PY 2000 VL 343 IS 23 BP 1703 EP 1714 DI 10.1056/NEJM200012073432307 PG 12 WC Medicine, General & Internal SC General & Internal Medicine GA 380PX UT WOS:000165711900007 PM 11106721 ER PT J AU Kim, JS Pirnia, F Choi, YH Nguyen, P Knepper, B Tsokos, M Schulte, TW Birrer, MJ Blagosklonny, MV Schaefer, O Mushinski, JF Trepel, JB AF Kim, JS Pirnia, F Choi, YH Nguyen, P Knepper, B Tsokos, M Schulte, TW Birrer, MJ Blagosklonny, MV Schaefer, O Mushinski, JF Trepel, JB TI Lovastatin induces apoptosis in a primitive neuroectodermal tumor cell line in association with RB down-regulation and loss of the G-1 checkpoint SO ONCOGENE LA English DT Article DE lovastatin; differentiation; apoptosis; ESFT ID PROSTATE CARCINOMA-CELLS; CYCLIN-DEPENDENT KINASES; EWINGS-SARCOMA; THYMOCYTE APOPTOSIS; DNA-BINDING; PHASE-I; CANCER; E2F-1; INHIBITION; INDUCTION AB To develop a new approach to the treatment of primitive neuroectodermal tumors we evaluated the effect of the HMG-CoA reductase inhibitor lovastatin on the Ewing's sarcoma cell line CHP-100, Lovastatin induced neural morphology and markers including neuron-specific enolase and neurofilament protein. The acquisition of neural morphology required nem mRNA synthesis, and cDNA microarray analysis confirmed that lovastatin altered the program of gene expression. After morphologic differentiation the cells underwent rapidly progressive apoptosis. In normal development of neuronal progenitors, differentiation signals trigger p21(WAF1) accumulation, RB hypophosphorylation, enhanced RB-E2F-1 association, and G1 arrest, and these events have been shown to protect from apoptosis. In contrast, in the Ewing's sarcoma cells lovastatin triggered differentiation without causing cell cycle arrest: p21(WAF1) was not induced, RE remained hyperphosphorylated, and RE protein expression and RE-E2F-1 association were markedly downregulated, suggesting that loss of an RE-regulated G1 checkpoint promoted apoptosis. Consistent with this hypothesis, adenoviral p21(WAF1) decreased DNA synthesis and partially protected from lovastatin-induced cytotoxicity. The data demonstrate a new model for examining the genetic regulation of cell fate in a neural progenitor tumor and suggest a new approach to the treatment of this neoplasm. C1 NCI, Med Branch, NIH, Bethesda, MD 20892 USA. NCI, Pathol Lab, Div Clin Sci, NIH, Bethesda, MD 20892 USA. NCI, Genet Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Trepel, JB (reprint author), NCI, Med Branch, NIH, Bldg 10,Room 12N230, Bethesda, MD 20892 USA. NR 58 TC 25 Z9 27 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD DEC 7 PY 2000 VL 19 IS 52 BP 6082 EP 6090 DI 10.1038/sj.onc.1204008 PG 9 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 382MY UT WOS:000165827900016 PM 11146561 ER PT J AU Shen, JN Wenger, N Glaspy, J Hays, RD Albert, PS Choi, C Shekelle, PG AF Shen, JN Wenger, N Glaspy, J Hays, RD Albert, PS Choi, C Shekelle, PG TI Electroacupuncture for control of myeloablative chemotherapy-induced emesis - A randomized controlled trial SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID CANCER-CHEMOTHERAPY; SUBSTANCE-P; ACUPUNCTURE; CISPLATIN; ANTIEMETICS; CYCLOPHOSPHAMIDE; ONDANSETRON; ENKEPHALIN; THERAPY; NAUSEA AB Context High-dose chemotherapy poses considerable challenges to emesis management. Although prior studies suggest that acupuncture may reduce nausea and emesis, it is unclear whether such benefit comes from the nonspecific effects of attention and clinician-patient interaction. Objective To compare the effectiveness of electroacupuncture vs minimal needling and mock electrical stimulation or antiemetic medications alone in controlling emesis among patients undergoing a highly emetogenic chemotherapy regimen. Design Three-arm, parallel-group, randomized controlled trial conducted from March 1996 to December 1997, with a 5-day study period and a 9-day follow-up. Setting Oncology center at a university medical center. Patients One hundred four women (mean age, 46 years) with high-risk breast cancer. Interventions Patients were randomly assigned to receive low-frequency electroacupuncture at classic antiemetic acupuncture points once daily for 5 days (n=37), minimal needling at control points with mock electrostimulation on the same schedule (n=33); or no adjunct needling (n=34). All patients received concurrent triple antiemetic pharmacotherapy and high-dose chemotherapy (cyclophosphamide, cisplatin, and carmustine). Main Outcome Measures Total number of emesis episodes occurring during the 5-day study period and the proportion of emesis-free days, compared among the 3 groups. Results The number of emesis episodes occurring during the 5 days was lower for patients receiving electroacupuncture compared with those receiving minimal needling or pharmacotherapy alone (median number of episodes, 5, 10, and 15, respectively; P<.001). The electroacupuncture group had fewer episodes of emesis than the minimal needling group (P<.001), whereas the minimal needling group had fewer episodes of emesis than the antiemetic pharmacotherapy alone group (P=.01), The differences among groups were not significant during the 9-day follow-up period (P=.18), Conclusions in this study of patients with breast cancer receiving high-dose chemotherapy, adjunct electroacupuncture was more effective in controlling emesis than minimal needling or antiemetic pharmacotherapy alone, although the observed effect had limited duration. C1 NIAAA, Clin Studies Lab, Brain Electrophysiol & Imaging Sect, NIH, Bethesda, MD 20892 USA. Univ Calif Los Angeles, Sch Med, Dept Med, Div Gen Internal Med & Hlth Serv Res, Los Angeles, CA 90024 USA. Univ Calif Los Angeles, Sch Med, Dept Med, Div Hematol & Oncol, Los Angeles, CA 90024 USA. RAND Corp, Santa Monica, CA USA. NCI, Biometr Res Branch, NIH, Bethesda, MD 20892 USA. W Los Angeles Vet Affairs Med Ctr, Dept Med, Vet Affairs Hlth Serv REs & Dev Serv, Los Angeles, CA 90073 USA. RP Shen, JN (reprint author), NIAAA, Clin Studies Lab, Brain Electrophysiol & Imaging Sect, NIH, Room 6 S-240,Mail Stop 1610,10 Ctr Dr,Bldg 10, Bethesda, MD 20892 USA. RI Hays, Ronald/D-5629-2013 FU BHP HRSA HHS [PE19001-09] NR 31 TC 149 Z9 158 U1 3 U2 9 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60654-0946 USA SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD DEC 6 PY 2000 VL 284 IS 21 BP 2755 EP 2761 DI 10.1001/jama.284.21.2755 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 377JB UT WOS:000165509500030 PM 11105182 ER PT J AU Brady, RO Schiffmann, R AF Brady, RO Schiffmann, R TI Clinical features of and recent advances in therapy for Fabry disease SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID INHERITED ENZYME DEFICIENCY; REPLACEMENT THERAPY; ALPHA-GALACTOSIDASE; GENE-TRANSFER; CELLS; MICE; DEFECTS AB Fabry disease is an X-linked recessive lysosomal storage disorder caused by a deficiency of alpha -galactosidase A. Intracellular accumulation of globotriaosylceramide, the glycolipid substrate of this enzyme, leads to severe painful neuropathy with progressive renal, cardiovascular, and cerebrovascular dysfunction and early death, Men are predominantly affected but many female carriers have similar clinical involvement, including increased risk of stroke. Physical stigmata, such as angiokeratomas in skin and mucous membranes and characteristic benign corneal abnormalities, facilitate identification of Fabry disease. The finding of a marked decreased activity of alpha -galactosidase A in white blood cells or cultured skin fibroblasts confirms the diagnosis. Treatment thus far has been symptomatic only, Etiology-based therapies are being developed that include enzyme replacement therapy, gene therapy, and substrate deprivation, Our recently completed double-blind, placebo-controlled trial of intravenous infusions of alpha -galactosidase A in patients with Fabry disease demonstrated the safety and efficacy of this treatment. C1 NINDS, Dev Metab Neurol Branch, NIH, Bethesda, MD 20892 USA. RP Schiffmann, R (reprint author), NINDS, Dev Metab Neurol Branch, NIH, Bldg 10,Room 3D03,10 Ctr Dr,MSC 1260, Bethesda, MD 20892 USA. NR 38 TC 122 Z9 131 U1 0 U2 2 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD DEC 6 PY 2000 VL 284 IS 21 BP 2771 EP 2775 DI 10.1001/jama.284.21.2771 PG 5 WC Medicine, General & Internal SC General & Internal Medicine GA 377JB UT WOS:000165509500032 PM 11105184 ER PT J AU Kerlikowske, K Carney, PA Geller, B Mandelson, MT Taplin, SH Malvin, K Ernster, V Urban, N Cutter, G Rosenberg, R Ballard-Barbash, R AF Kerlikowske, K Carney, PA Geller, B Mandelson, MT Taplin, SH Malvin, K Ernster, V Urban, N Cutter, G Rosenberg, R Ballard-Barbash, R TI Performance of screening mammography among women with and without a first-degree relative with breast cancer SO ANNALS OF INTERNAL MEDICINE LA English DT Article ID POSITIVE PREDICTIVE VALUE; FAMILY HISTORY; GROWTH-RATES; FOLLOW-UP; AGE; RISK; SENSITIVITY; DATABASE; INTERVAL; DENSITY AB Background: Although it is recommended that women with a family history of breast cancer begin screening mammography at a younger age than average-risk women, few studies have evaluated the performance of mammography in this group. Objective: To compare the performance of screening mammography in women with a first-degree family history of breast cancer and women of similar age without such history. Design: Cross-sectional. Setting: Mammography registries in California (n = 1) New Hampshire (n = 1), New Mexico (n = 1), Vermont (n = 1) Washington State (n = 2), and Colorado (n = 1). Measurements: Risk factors for breast cancer; results of first screening examination captured for a woman by a registry; and any invasive cancer or ductal carcinoma in situ identified by linkage to a pathology database, the Surveillance, Epidemiology, and End Results program, or a state tumor registry. Results: The number of cancer cases per 1000 examinations increased with age and was higher in women with a family history of breast cancer than in those without (3.2 vs. 1.6 for ages 30 to 39 years, 4.7 vs. 2.7 for ages 40 to 49 years, 6.6 vs. 4.6 for ages 50 to 59 years, and 9.3 vs. 6.9 for ages 60 to 69 years). The sensitivity of mammography increased significantly with age (P = 0.001 [chi-square test for trend]) in women with a family history and in those without (63.2% [95% CI, 41.5% to 84.8%] vs. 69.5% [Ct, 57.7% to 81.2%] for ages 30 to 39 years, 70.2% [CI, 61.0% to 79.5%] vs. 77.5% [CI, 73.3% to 81.8%] for ages 40 to 49 years, 81.3% [CI, 73.3% to 89.3%] vs. 80.2% [CI, 76.5% to 83.9%] for ages 50 to 59 years, and 83.8% [CI, 76.8% to 90.9%] vs. 87.7% [Cf, 84.8% to 90.7%] for ages 60 to 69 years). Sensitivity was similar for each decade of age regardless of family history. The positive predictive value of mammography was higher in women with a family history than in those without (3.7% vs. 2.9%; P = 0.001). Conclusions: Cancer detection rates in women who had a first-degree relative with a history of breast cancer were similar to those in women a decade older without such a history. The sensitivity of screening mammography was influenced primarily by age. C1 Univ Calif San Francisco, San Francisco, CA 94143 USA. Dartmouth Med Sch, Lebanon, NH USA. Univ Vermont, Coll Med, Burlington, VT USA. Grp Hlth Cooperat Puget Sound, Seattle, WA 98121 USA. Fred Hutchinson Canc Res Ctr, Seattle, WA 98104 USA. Ctr Canc Res, AMC, Denver, CO USA. Univ New Mexico, Med Ctr, New Mexico Tumor Registry, Albuquerque, NM 87131 USA. Natl Canc Inst, Bethesda, MD USA. RP Kerlikowske, K (reprint author), Vet Affairs Med Ctr, Gen Internal Med Sect, 111A1,4150 Clement St, San Francisco, CA 94121 USA. FU NCI NIH HHS [CA63146, 1 U01 CA 63740, K07CA71869] NR 39 TC 130 Z9 132 U1 0 U2 5 PU AMER COLL PHYSICIANS PI PHILADELPHIA PA INDEPENDENCE MALL WEST 6TH AND RACE ST, PHILADELPHIA, PA 19106-1572 USA SN 0003-4819 EI 1539-3704 J9 ANN INTERN MED JI Ann. Intern. Med. PD DEC 5 PY 2000 VL 133 IS 11 BP 855 EP 863 PG 9 WC Medicine, General & Internal SC General & Internal Medicine GA 378LL UT WOS:000165585800004 PM 11103055 ER PT J AU Brown, ML Hankey, BF Ballard-Barbash, R AF Brown, ML Hankey, BF Ballard-Barbash, R TI Measuring the quality of breast cancer care SO ANNALS OF INTERNAL MEDICINE LA English DT Letter ID PATTERNS; WOMEN C1 NCI, Bethesda, MD 20892 USA. RP Brown, ML (reprint author), NCI, Bethesda, MD 20892 USA. NR 5 TC 13 Z9 13 U1 1 U2 1 PU AMER COLL PHYSICIANS PI PHILADELPHIA PA INDEPENDENCE MALL WEST 6TH AND RACE ST, PHILADELPHIA, PA 19106-1572 USA SN 0003-4819 EI 1539-3704 J9 ANN INTERN MED JI Ann. Intern. Med. PD DEC 5 PY 2000 VL 133 IS 11 BP 920 EP 920 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 378LL UT WOS:000165585800011 PM 11103066 ER PT J AU Natarajan, K Sawicki, MW Margulies, DH Mariuzza, RA AF Natarajan, K Sawicki, MW Margulies, DH Mariuzza, RA TI Crystal structure of human CD69: A C-type lectin-like activation marker of hematopoietic cells SO BIOCHEMISTRY LA English DT Article ID LYMPHOCYTE-ACTIVATION; INDUCER MOLECULE; ANTIGEN CD69; T-CELLS; RECEPTORS; SUPERFAMILY; EXPRESSION; DOMAIN; IDENTIFICATION; PROTEINS AB CD69 is a widely expressed type II transmembrane glycoprotein related to the C-type animal lectins that exhibits regulated expression on a variety of cells of the hematopoietic lineage, including neutrophils, monocytes, T cells, B cells, natural killer (NK) cells, and platelets. Activation of T lymphocytes results in the induced expression of CD69 at the cell surface. In addition, cross-linking of CD69 by specific antibodies leads to the activation of cells bearing this receptor and to the induction of effector functions. However, the physiological ligand of CD69 is unknown. We report here the X-ray crystal structure of the extracellular C-type lectin-like domain (CTLD) of human CD69 at 2.27 Angstrom resolution. Recombinant CD69 was expressed in bacterial inclusion bodies and folded in vitro. The protein, which exists as a disulfide-linked homodimer on the cell surface, crystallizes as a symmetrical dimer, similar to those formed by the related NK cell receptors Ly49A and CD94. The structure reveals conservation of the C-type lectin-like fold, including preservation of the two alpha -helical regions found in Ly49A and mannose-binding protein (MBP). However, only one of the nine residues coordinated to Ca2+ in MBP is conserved in CD69 and no bound Ca2+ is evident in the crystal structure. Surprisingly, electron density suggestive of a puckered six-membered ring was discovered at a site structurally analogous to the ligand-binding sites of MBP and Ly49A. This sugar-like density may represent, or mimic, part of the natural ligand recognized by CD69. C1 NIAID, Mol Biol Sect, Immunol Lab, NIH, Bethesda, MD 20892 USA. Univ Maryland, Inst Biotechnol, Ctr Adv Res Biotechnol, Rockville, MD 20850 USA. RP Mariuzza, RA (reprint author), NIAID, Mol Biol Sect, Immunol Lab, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. RI Margulies, David/H-7089-2013; OI Margulies, David/0000-0001-8530-7375 FU NIAID NIH HHS [AI47990] NR 45 TC 54 Z9 57 U1 0 U2 0 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD DEC 5 PY 2000 VL 39 IS 48 BP 14779 EP 14786 DI 10.1021/bi0018180 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 378XF UT WOS:000165610100009 PM 11101293 ER PT J AU Rothman, RB Baumann, MH Savage, JE Rauser, L McBride, A Hufeisen, SJ Roth, BL AF Rothman, RB Baumann, MH Savage, JE Rauser, L McBride, A Hufeisen, SJ Roth, BL TI Evidence for possible involvement of 5-HT2B receptors in the cardiac valvulopathy associated with fenfluramine and other serotonergic medications SO CIRCULATION LA English DT Article DE valves; fenfluramine; norfenfluramine; receptors ID VALVULAR HEART-DISEASE; APPETITE-SUPPRESSANT DRUGS; 5-HYDROXYTRYPTAMINE(2A) RECEPTORS; AGONIST BINDING; OBESE PATIENTS; ECHOCARDIOGRAPHY; DEXFENFLURAMINE; PHARMACOKINETICS; ABNORMALITIES; PHENTERMINE AB Background-Serotonergic medications with various mechanisms of action are used to treat psychiatric disorders and are being investigated as treatments for drug dependence. The occurrence of fenfluramine-associated valvular heart disease (VHD) has raised concerns that other serotonergic medications might also increase the risk of developing VHD. We hypothesized that fenfluramine or its metabolite norfenfluramine and other medications known to produce VHD have preferentially high affinities for a particular serotonin receptor subtype capable of stimulating mitogenesis. Methods and Results-Medications known or suspected to cause VHD (positive controls) and medications not associated with VHD (negative controls) were screened for activity at 11 cloned serotonin receptor subtypes by use of ligand-binding methods and functional assays. The positive control drugs were (+/-)-fenfluramine; (+)-fenfluramine; (-)-fenfluramine; its metabolites (+/-)-norfenfluramine, (+)-norfenfluramine, and (-)-norfenfluramine; ergotamine; and methysergide and its metabolite methylergonovine. The negative control drugs were phentermine, fluoxetine, its metabolite norfluoxetine, and trazodone and its active metabolite m-chlorophenylpiperazine. (+/-)-, (+)-, and (-)-Norfenfluramine, ergotamine, and methylergonovine all had preferentially high affinities for the cloned human serotonin 5-HT2B receptor and were partial to full agonists at the 5-HT2B receptor. Conclusions-Our data imply that activation of 5-HT2B receptors is necessary to produce VHD and that serotonergic medications that do not activate 5-HT2B receptors are unlikely to produce VHD. We suggest that all clinically available medications with serotonergic activity and their active metabolites be screened for agonist activity at 5-HT2B receptors and that clinicians should consider suspending their use of medications with significant activity at 5-HT2B receptors. C1 NIDA, DIR, CPS, NIH, Baltimore, MD 21224 USA. Case Western Reserve Univ, Dept Biochem, Cleveland, OH 44106 USA. Case Western Reserve Univ, Dept Psychiat, Cleveland, OH 44106 USA. Case Western Reserve Univ, Dept Neurosci, Cleveland, OH 44106 USA. Case Western Reserve Univ, NIMH, Psychoact Drug Screening Program, Cleveland, OH 44106 USA. RP Rothman, RB (reprint author), NIDA, DIR, CPS, NIH, POB 5180,5500 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Roth, Bryan/F-3928-2010 FU NIMH NIH HHS [K02MH01367, N01MH80005] NR 33 TC 382 Z9 389 U1 2 U2 12 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD DEC 5 PY 2000 VL 102 IS 23 BP 2836 EP 2841 PG 6 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 378VZ UT WOS:000165607000008 PM 11104741 ER PT J AU Gronthos, S Mankani, M Brahim, J Robey, PG Shi, S AF Gronthos, S Mankani, M Brahim, J Robey, PG Shi, S TI Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE odontoblast; dentin; in vivo transplantation ID MARROW STROMAL CELLS; GROWTH-FACTOR; SEQUENCE DETERMINATION; ALKALINE-PHOSPHATASE; GENE-EXPRESSION; IN-VIVO; BONE; DIFFERENTIATION; PROLIFERATION; SIALOPROTEIN AB Dentinal repair in the postnatal organism occurs through the activity of specialized cells, odontoblasts, that are thought to be maintained by an as yet undefined precursor population associated with pulp tissue. In this study, we isolated a clonogenic, rapidly proliferative population of cells from adult human dental pulp. These DPSCs were then compared with human bone marrow stromal cells (BMSCs), known precursors of osteoblasts. Although they share a similar immunophenotype in vitro, functional studies showed that DPSCs produced only sporadic, but densely calcified nodules, and did not form adipocytes, whereas BMSCs routinely calcified throughout the adherent cell layer with dusters of lipid-laden adipocytes. When DPSCs were transplanted into immunocompromised mice, they generated a dentin-like structure lined with human odontoblast-like cells that surrounded a pulp-like interstitial tissue. in contrast, BMSCs formed lamellar bone containing osteocytes and surface-lining osteoblasts, surrounding a fibrous vascular tissue with active hematopoiesis and adipocytes. This study isolates postnatal human DPSCs that have the ability to form a dentin/pulp-like complex. C1 NIDR, Craniofacial & Skeletal Dis Branch, NIH, Bethesda, MD 20892 USA. RP Shi, S (reprint author), NIDR, Craniofacial & Skeletal Dis Branch, NIH, Bethesda, MD 20892 USA. RI Robey, Pamela/H-1429-2011 OI Robey, Pamela/0000-0002-5316-5576 NR 39 TC 1613 Z9 1770 U1 17 U2 137 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD DEC 5 PY 2000 VL 97 IS 25 BP 13625 EP 13630 DI 10.1073/pnas.240309797 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 380XH UT WOS:000165728800033 PM 11087820 ER PT J AU Lempicki, RA Kovacs, JA Baseler, MW Adelsberger, JW Dewar, RL Natarajan, V Bosche, MC Metcalf, JA Stevens, RA Lambert, LA Alvord, WG Polis, MA Davey, RT Dimitrov, DS Lane, HC AF Lempicki, RA Kovacs, JA Baseler, MW Adelsberger, JW Dewar, RL Natarajan, V Bosche, MC Metcalf, JA Stevens, RA Lambert, LA Alvord, WG Polis, MA Davey, RT Dimitrov, DS Lane, HC TI Impact of HIV-1 infection and highly active antiretroviral therapy on the kinetics of CD4(+) and CD8(+) T cell turnover in HIV-infected patients SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE AIDS; proliferation; immune activation; T cell receptor rearrangement excision circles ID RAPID TURNOVER; LYMPHOCYTES; MACAQUES; ANTIGEN; COUNTS AB To evaluate the effects of HIV infection on T cell turnover, we examined levels of DNA synthesis in lymph node and peripheral blood mononuclear cell subsets by using ex vivo labeling with BrdUrd. Compared with healthy controls (n = 67), HIV-infected patients (n = 57) had significant increases in the number and fraction of dividing CD4(+) and CD8(+) T cells. Higher percentages of dividing CD4+ and CD8(+) T cells were noted in patients with the higher viral burdens. No direct correlation was noted between rates of T cell turnover and CD4(+) T cell counts. Marked reductions in CD4+ and CD8(+) T cell proliferation were seen in 11/11 patients 1-12 weeks after initiation of highly active antiretroviral therapy (HAART). These reductions persisted for the length of the study (16-72 weeks). Decreases in naive T cell proliferation correlated with increases in the levels of T cell receptor rearrangement excision circles. Division of CD4(+) and CD8(+) T cells increased dramatically in association with rapid increases in HIV-1 viral loads in 9/9 patients 5 weeks after termination of HAART and declined to pre-HAART-termination levels 8 weeks after reinitiation of therapy. These data are consistent with the hypothesis that HIV-1 infection induces a viral burden-related, global activation of the immune system, leading to increases in lymphocyte proliferation. C1 NIAID, Lab Immunoregulat, Clin & Mol Retrovirol Sect, NIH, Bethesda, MD 20892 USA. Sci Applicat Int Corp, Clin Serv Program, Frederick, MD 21702 USA. Data Management Serv Inc, Comp Serv, Frederick, MD 21702 USA. Data Management Serv Inc, Stat Serv, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, Lab Expt & Computat Biol, Frederick, MD 21702 USA. NIAID, Ctr Clin, Dept Crit Care Med, NIH, Bethesda, MD 20892 USA. NIAID, Lab Immunoregulat, Clin & Mol Retrovirol Sect, NIH, Bethesda, MD 20892 USA. RP Lane, HC (reprint author), NIAID, Lab Immunoregulat, Clin & Mol Retrovirol Sect, NIH, Bldg 10,Room 11S231, Bethesda, MD 20892 USA. RI Lempicki, Richard/E-1844-2012; OI Lempicki, Richard/0000-0002-7059-409X; Polis, Michael/0000-0002-9151-2268 FU PHS HHS [N01-C0-56000] NR 24 TC 136 Z9 142 U1 0 U2 0 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD DEC 5 PY 2000 VL 97 IS 25 BP 13778 EP 13783 DI 10.1073/pnas.250472097 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 380XH UT WOS:000165728800059 PM 11095734 ER PT J AU Cocchi, F DeVico, AL Yarchoan, R Redfield, R Cleghorn, F Blattner, WA Garzino-Demo, A Colombino-Hatch, S Margolis, D Gallo, RC AF Cocchi, F DeVico, AL Yarchoan, R Redfield, R Cleghorn, F Blattner, WA Garzino-Demo, A Colombino-Hatch, S Margolis, D Gallo, RC TI Higher macrophage inflammatory protein (MIP)-1 alpha and MIP-1 beta levels from CD8(+) T cells are associated with asymptomatic HIV-1 infection SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID IMMUNODEFICIENCY-VIRUS-INFECTION; PERIPHERAL-BLOOD LYMPHOCYTES; SERUM CHEMOKINE LEVELS; BETA-CHEMOKINES; DISEASE PROGRESSION; TYPE-1 INFECTION; LONGITUDINAL ANALYSIS; CLINICAL COURSE; VIRAL LOAD; REPLICATION AB To test the hypothesis that beta -chemokine levels may be relevant to the control of HN in vivo, we compared RANTES, MIP-1 alpha, and MIP-1 beta production from purified CD8(+) T cells from 81 HIV-infected subjects and from 28 uninfected donors. Asymptomatic HIV+ subjects produced significantly higher levels of MIP-1 alpha and MIP-1 beta, but not RANTES, than uninfected donors or patients that progressed to AIDS. In contrast, beta chemokines in plasma were either nondetectable or showed no correlation with clinical status. The high beta -chemokine-mediated anti-HIV activity was against the macrophage tropic isolate HIV-1(BAL), with no demonstrable effect on the replication of the T-cell tropic HIV-1(IIIB) These findings suggest that constitutive beta -chemokine production may play an important role in the outcome of HIV-1 infection. C1 Univ Maryland, Maryland Biotechnol Inst, Inst Human Virol, Baltimore, MD 21201 USA. NCI, HIV & AIDS Malignancy Branch, NIH, Bethesda, MD 20892 USA. RP Cocchi, F (reprint author), Univ Maryland, Maryland Biotechnol Inst, Inst Human Virol, 725 W Lombard St, Baltimore, MD 21201 USA. OI Margolis, David/0000-0001-5714-0002 FU NIAID NIH HHS [R21AI44735] NR 61 TC 88 Z9 95 U1 0 U2 2 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD DEC 5 PY 2000 VL 97 IS 25 BP 13812 EP 13817 DI 10.1073/pnas.240469997 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 380XH UT WOS:000165728800065 PM 11095721 ER PT J AU Ott, DE Coren, LV Chertova, EN Gagliardi, TD Schubert, U AF Ott, DE Coren, LV Chertova, EN Gagliardi, TD Schubert, U TI Ubiquitination of HIV-1 and MuLV Gag SO VIROLOGY LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; MURINE LEUKEMIA-VIRUS; GENOMIC RNA; ZINC-FINGER; PROTEIN; TYPE-1; P6(GAG); DOMAIN; DEGRADATION; PROTEASOME AB Our previous biochemical studies of HIV-1 and MuLV virions isolated and identified mature Gag products, HIV-1 p6(Gag) and MuLV p12(Gag), that were conjugated to a single ubiquitin. To study the importance of the monoubiquitination of Gag, a series of lysine to arginine mutants were constructed that eliminated ubiquitination at one or both of the lysines in HIV-1(NL4-3) p6(Gag) and both lysines in Moloney MuLV p12(Gag). HPLC and immunoblot analysis of the HIV-1 mutants demonstrated that either of the lysines in p6(Gag), K27 or K33, could be monoubiquitinated. However, infectivity assays showed that monoubiquitination of HIV-1 p6(Gag) or MuLV p12(Gag) is not required for viral replication in vitro. Pulse-chase radiolabeling of HIV-1-producing cells revealed that monoubiquitination of p6(Gag) does not affect the short-term release of Virus from the cell, the maturation or Pr55(Gag), Or the sensitivity of these processes to proteasome inhibitors. Experiments with protease-deficient HIV-1 showed that Pr55(Gag) can be monoubiquitinated, suggesting that p6(Gag) is first modified as a domain within Gag. Examination of the proteins inside an HIV-1 mutant found that free ubiquitin was incorporated into the virions in the absence of the lysines in p6(Gag), showing that the ubiquitin inside the virus is not initially brought in as a p6(Gag) conjugate. Although our results establish that monoubiquitination of p6(Gag) and p12(Gag) is not required for Viral replication in vitro, this modification may be a by-product of interactions between Gag and cellular proteins during assembly and budding, (C) 2000 Academic Press. C1 NCI, SAIC, AIDS Vaccine Program, FCRDC, Frederick, MD 21702 USA. NIAID, Viral Dis Lab, NIH, Bethesda, MD 20892 USA. Heinrich Pette Inst Expt Virol & Immunol, D-20251 Hamburg, Germany. RP Ott, DE (reprint author), NCI, SAIC, AIDS Vaccine Program, FCRDC, POB B, Frederick, MD 21702 USA. FU NCI NIH HHS [N01CO56000] NR 48 TC 110 Z9 111 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0042-6822 J9 VIROLOGY JI Virology PD DEC 5 PY 2000 VL 278 IS 1 BP 111 EP 121 DI 10.1006/viro.2000.0648 PG 11 WC Virology SC Virology GA 383QU UT WOS:000165894500014 PM 11112487 ER PT J AU Senkevich, TG Weisberg, AS Moss, B AF Senkevich, TG Weisberg, AS Moss, B TI Vaccinia virus E10R protein is associated with the membranes of intracellular mature virions and has a role in morphogenesis SO VIROLOGY LA English DT Article ID COLI LAC REPRESSOR; LIVER-REGENERATION; GENE; EXPRESSION; CLONING; GENOME AB This study provides the initial biochemical, microscopic, and genetic characterization of the product of the vaccinia virus E10R gene, which belongs to the ERV1/ALR family of eukaryotic proteins, is conserved in all poxviruses and has homologs in other cytoplasmic DNA viruses. DNA encoding a short epitope tag was appended to the C-terminus of the 95-amino-acid open-reading frame without affecting virus reproduction. The E10R protein was synthesized after DNA replication and was associated with purified intracellular mature virions (IMV), from which it could be extracted with a nonionic detergent. Antibody to the tag decorated the surface of IMV, consistent with the anchorage of the E10R protein to the membrane via its hydrophobic N-terminus. Immunoelectron microscopy revealed that the E10R protein was associated with crescent membranes, immature virions, IMV, and extracellular particles. To investigate the role of E10R in the virus life cycle, we constructed an inducer-dependent null mutant. In the absence of inducer, the formation of infectious virus was severely inhibited and electron microscopy revealed an assembly block with accumulation of crescent membranes and immature virions. Cysteines 43 and 46, comprising a putative redox motif common to all poxvirus E10R homologs, were essential for complementation of the mutant virus by transfected E10R DNA. (C) 2000 Academic Press. C1 NIAID, Viral Dis Lab, NIH, Bethesda, MD 20892 USA. RP Moss, B (reprint author), NIAID, Viral Dis Lab, NIH, 4 Ctr Dr,MSC 0445, Bethesda, MD 20892 USA. NR 20 TC 51 Z9 52 U1 1 U2 5 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0042-6822 J9 VIROLOGY JI Virology PD DEC 5 PY 2000 VL 278 IS 1 BP 244 EP 252 DI 10.1006/viro.2000.0656 PG 9 WC Virology SC Virology GA 383QU UT WOS:000165894500026 PM 11112499 ER PT J AU Yan, WL Guan, XY Green, ED Nicolson, R Yap, TK Zhang, JH Jacobsen, LK Krasnewich, DM Kumra, S Lenane, MC Gochman, P Damschroder-Williams, PJ Esterling, LE Long, RT Martin, BM Sidransky, E Rapoport, JL Ginns, EI AF Yan, WL Guan, XY Green, ED Nicolson, R Yap, TK Zhang, JH Jacobsen, LK Krasnewich, DM Kumra, S Lenane, MC Gochman, P Damschroder-Williams, PJ Esterling, LE Long, RT Martin, BM Sidransky, E Rapoport, JL Ginns, EI TI Childhood-onset schizophrenia/autistic disorder and t(1;7) reciprocal translocation: Identification of a BAC contig spanning the translocation breakpoint at 7q21 SO AMERICAN JOURNAL OF MEDICAL GENETICS LA English DT Article DE psychosis; autistic disorder; translocation ID SUSCEPTIBILITY GENES; AUTOIMMUNE-DISEASES; AUTISTIC DISORDER; ASSOCIATION; CHROMOSOME-7; PEDIGREES; LINKAGE; SPEECH; STATE; SCAN AB Childhood-onset schizophrenia (COS) is defined by the development of first psychotic symptoms by age 12, While recruiting patients with COS refractory to conventional treatments for a trial of atypical antipsychotic drugs, we discovered a unique case who has a familial t(1;7)(p22;q21) reciprocal translocation and onset of psychosis at age 9, The patient also has symptoms of autistic disorder, which are usually transient before the first psychotic episode among 40-50% of the childhood schizophrenics but has persisted in him even after the remission of psychosis, Cosegregating with the translocation, among the carriers in the family available for the study, are other significant psychopathologies, including alcohol/drug abuse, severe impulsivity, and paranoid personality and language delay, This case may provide a model for understanding the genetic basis of schizophrenia or autism. Here we report the progress toward characterization of genomic organization across the translocation breakpoint at 7q21, The polymorphic markers, D7S630/D7S492 and D7S2410/D7S646, immediately flanking the breakpoint, may be useful for further confirming the genetic linkage for schizophrenia or autism in this region. Published 2000 Wiley-Liss, Inc.(dagger). C1 NIMH, Child Psychiat Branch, Bethesda, MD 20892 USA. NIMH, Clin Neurosci Branch, Bethesda, MD 20892 USA. Natl Human Genom Res Inst, Canc Genet Lab, Bethesda, MD USA. Natl Human Genom Res Inst, Genome Technol Branch, Bethesda, MD USA. NIH, Div Comp Res & Technol, Bethesda, MD 20892 USA. Natl Lib Med, Natl Ctr Biotechnol Informat, Bethesda, MD 20894 USA. Natl Inst Deafness & Other Commun Disorders, Genet Mol Lab, Bethesda, MD USA. RP Yan, WL (reprint author), NIMH, Child Psychiat Branch, Bldg 49,Rm B1EE16, Bethesda, MD 20892 USA. RI Nicolson, Robert/E-4797-2011; Guan, Xin-Yuan/A-3639-2009 OI Guan, Xin-Yuan/0000-0002-4485-6017 NR 31 TC 34 Z9 34 U1 3 U2 6 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0148-7299 J9 AM J MED GENET JI Am. J. Med. Genet. PD DEC 4 PY 2000 VL 96 IS 6 BP 749 EP 753 DI 10.1002/1096-8628(20001204)96:6<749::AID-AJMG10>3.0.CO;2-K PG 5 WC Genetics & Heredity SC Genetics & Heredity GA 380RM UT WOS:000165717300010 PM 11121174 ER PT J AU Rosenberg, RD Yankaskas, BC Hunt, WC Ballard-Barbash, R Urban, N Ernster, VL Kerlikowske, K Geller, B Carney, PA Taplin, S AF Rosenberg, RD Yankaskas, BC Hunt, WC Ballard-Barbash, R Urban, N Ernster, VL Kerlikowske, K Geller, B Carney, PA Taplin, S TI Effect of variations in operational definitions on performance estimates for screening mammography SO ACADEMIC RADIOLOGY LA English DT Article DE definitions; breast cancer; mass screening; methods; sensitivity; specificity ID ESTROGEN REPLACEMENT THERAPY; POSITIVE PREDICTIVE VALUE; BREAST-CANCER; FAMILY HISTORY; NEW-MEXICO; SENSITIVITY; AGE; SPECIFICITY; ALBUQUERQUE; COMMUNITY AB Rationale and Objectives. The Mammography Quality Standards Act requires practices to measure limited aspects of their performance. The authors conducted this study to calculate the differences in measurements of sensitivity and specificity due only to differences in the definitions used in the analysis. This included definitions for case inclusion. Materials and Methods. Data from the New Mexico Mammography Project for January 1991 to December 1995 on 136,540 women who underwent screening mammography were analyzed. A starting definition was created for each performance measure. The components of the definition were varied, and estimates of sensitivity and specificity for the different definitions were calculated. Results, Sensitivity was lower and specificity was higher when assessed on the basis of the results of all imaging performed in the screening work-up rather than on the initial screening examination alone. Sensitivity was higher and specificity was lower in women who did not undergo rather than in women who did recently undergo a previous examination. When the definition of a positive examination included cases that were recommended for short-term follow-up, the work-up sensitivity was slightly higher and the work-up specificity was considerably lower. Longer follow-up times for determining the diagnosis of cancer were associated with decreasing sensitivity, particularly when the follow-up period extended beyond 12 months. Conclusion. Variations in the operational definitions for measures of mammographic performance affect these estimates. To facilitate valid comparisons, reports need to be explicit regarding the definitions and methods used. C1 Univ New Mexico, Hlth Sci Ctr, Dept Radiol, Albuquerque, NM 87131 USA. Univ N Carolina, Dept Radiol, Chapel Hill, NC USA. Univ New Mexico, New Mexico Tumor Registry, Albuquerque, NM 87131 USA. NCI, Appl Res Branch, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. Fred Hutchinson Canc Res Ctr, Seattle, WA 98104 USA. Univ Calif San Francisco, Dept Epidemiol & Biostat, San Francisco, CA 94143 USA. Univ Calif San Francisco, Gen Internal Med Sect, San Francisco, CA 94143 USA. Univ Vermont, Off Hlth Promot Res, Burlington, VT 05405 USA. Dartmouth Coll, Hitchcock Med Ctr, Dartmouth Med Sch, New Hampshire Mammog Network, Hanover, NH 03756 USA. Dartmouth Coll, Hitchcock Med Ctr, Dartmouth Med Sch, Norris Cotton Canc Ctr, Hanover, NH 03756 USA. Grp Hlth Cooperat Puget Sound, Ctr Hlth Studies, Seattle, WA 98101 USA. RP Rosenberg, RD (reprint author), Univ New Mexico, Hlth Sci Ctr, Dept Radiol, 900 Camino de Salid NE, Albuquerque, NM 87131 USA. FU NCI NIH HHS [CA-95-004] NR 21 TC 26 Z9 26 U1 0 U2 0 PU ASSOC UNIV RADIOLOGISTS PI OAK BROOK PA 820 JORIE BLVD, OAK BROOK, IL 60523-2251 USA SN 1076-6332 J9 ACAD RADIOL JI Acad. Radiol. PD DEC PY 2000 VL 7 IS 12 BP 1058 EP 1068 DI 10.1016/S1076-6332(00)80057-4 PG 11 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 380RH UT WOS:000165716900002 PM 11131050 ER PT J AU Chen, YW Clore, GM AF Chen, YW Clore, GM TI A systematic case study on using NMR models for molecular replacement: p53 tetramerization domain revisited SO ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY LA English DT Article ID PROTEIN CRYSTAL-STRUCTURE; HIV-INACTIVATING PROTEIN; OLIGOMERIZATION DOMAIN; TUMOR-SUPPRESSOR; CYANOVIRIN-N; RESOLUTION; BINDING; PACKING; PACKAGE; AMORE AB Molecular replacement using search models derived from nuclear magnetic resonance (NMR) spectroscopy has often proved problematic. It has been known for some time that the overall differences in atomic positions (r.m.s.d.) between the crystalline and the solution states of the same protein are of the order of 1-2 Angstrom and approach the limit of molecular replacement. In most cases, this structural difference is a result of calculating the NMR structure with insufficient data, yielding an NMR structure of limited accuracy. A systematic case study was performed to investigate the use of NMR models for molecular replacement on the p53 tetramerization domain: NMR search models of varying degrees of accuracy were employed to solve phases for the 1.5 Angstrom X-ray diffraction data. An approximate correlation was found between the accuracy of the NMR search model and the clarity and quality of the molecular-replacement solution. It was found that ensemble models perform better than single averaged models and have a larger tolerance in model inaccuracy. Also, distance-derived B factors can improve the performance of single models. C1 Ctr Prot Engn, Cambridge CB2 2QH, England. Univ Cambridge, Chem Lab, MRC Ctr, Cambridge CB2 2QH, England. NIDDKD, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. RP Chen, YW (reprint author), Ctr Prot Engn, Hills Rd, Cambridge CB2 2QH, England. RI Clore, G. Marius/A-3511-2008 OI Clore, G. Marius/0000-0003-3809-1027 NR 28 TC 10 Z9 10 U1 0 U2 2 PU MUNKSGAARD INT PUBL LTD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0907-4449 J9 ACTA CRYSTALLOGR D JI Acta Crystallogr. Sect. D-Biol. Crystallogr. PD DEC PY 2000 VL 56 BP 1535 EP 1540 DI 10.1107/S0907444900012002 PN 12 PG 6 WC Biochemical Research Methods; Biochemistry & Molecular Biology; Biophysics; Crystallography SC Biochemistry & Molecular Biology; Biophysics; Crystallography GA 377HX UT WOS:000165509100002 PM 11092918 ER PT J AU Evdokimov, AG Anderson, DE Routzahn, KM Waugh, DS AF Evdokimov, AG Anderson, DE Routzahn, KM Waugh, DS TI Overproduction, purification, crystallization and preliminary X-ray diffraction analysis of YopM, an essential virulence factor extruded by the plague bacterium Yersinia pestis SO ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY LA English DT Article ID V-ANTIGEN; TYROSINE-PHOSPHATASE; BINDING PROTEIN; HOST-CELLS; EFFECTOR; VIRULON; KINASE AB A recombinant form of Yersinia pestis YopM with a C-terminal polyhistidine affinity tag has been overproduced in Escherichia coli, purified to homogeneity and crystallized using the hanging-drop vapor-diffusion technique. Several different crystal forms were obtained. The most suitable crystals for X-ray diffraction belonged to space groups P4(2)22 (unit-cell parameters a = 109.36, b = 109.36, c = 101.50 Angstrom) and C222(1) (unit-cell parameters a = 71.73, b = 121.85, c = 189.79 Angstrom). With a synchrotron-radiation source, these crystals diffracted to 2.4 and 1.9 Angstrom resolution, respectively. C1 NCI, Prot Engn Sect, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. NCI, Struct Biol Core Facil, Program Struct Biol, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RP Waugh, DS (reprint author), NCI, Prot Engn Sect, Frederick Canc Res & Dev Ctr, POB B, Frederick, MD 21702 USA. NR 21 TC 14 Z9 14 U1 0 U2 1 PU MUNKSGAARD INT PUBL LTD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0907-4449 J9 ACTA CRYSTALLOGR D JI Acta Crystallogr. Sect. D-Biol. Crystallogr. PD DEC PY 2000 VL 56 BP 1676 EP 1679 DI 10.1107/S0907444900013378 PN 12 PG 4 WC Biochemical Research Methods; Biochemistry & Molecular Biology; Biophysics; Crystallography SC Biochemistry & Molecular Biology; Biophysics; Crystallography GA 377HX UT WOS:000165509100028 PM 11092944 ER PT J AU Wong, K Armstrong, RC Gyure, KA Morrison, AL Rodriguez, D Matalon, R Johnson, AB Wollmann, R Gilbert, E Le, TQ Bradley, CA Crutchfield, K Schiffmann, R AF Wong, K Armstrong, RC Gyure, KA Morrison, AL Rodriguez, D Matalon, R Johnson, AB Wollmann, R Gilbert, E Le, TQ Bradley, CA Crutchfield, K Schiffmann, R TI Foamy cells with oligodendroglial phenotype in childhood ataxia with diffuse central nervous system hypomyelination syndrome SO ACTA NEUROPATHOLOGICA LA English DT Article DE childhood ataxia with diffuse central; nervous system hypomyelination (CACH syndrome); leukodystrophy; myelin; oligodendroglia ID MAGNETIC-RESONANCE SPECTROSCOPY; PELIZAEUS-MERZBACHER DISEASE; VANISHING WHITE-MATTER; MULTIPLE-SCLEROSIS; OPTIC-NERVE; MYELIN; CHILDREN; PROTEIN; MICE; LEUKOENCEPHALOPATHY AB Childhood ataxia with diffuse central nervous system hypomyelination syndrome (CACH) is a recently described leukodystrophy of unknown etiology. To characterize the neuropathological features and gain insight as to the pathogenesis of this disorder, we studied cerebral tissue from six patients with the CACH syndrome. Evaluation of toluidine blue-stained, semithin sections of white matter from CACH patients disclosed unusual cells with "foamy" cytoplasm, small round nuclei and fine chromatin. Electron microscopy (EM) revealed cells in the white matter with abundant cytoplasm containing many mitochondria and loosely clustered, membranous structures, but lacking the lysosomal structures seen in macrophages. Further analysis of tissue sections with antibodies and special stains demonstrated that the abnormal cells with abundant cytoplasm labeled with oligodendroglial markers, but did not react with macrophage or astrocytic markers. Double immunolabeling with macrophage and oligodendroglial markers clearly distinguished macrophages from the "foamy" oligodendroglial cells (FODCs). Proteolipid protein (PLP) mRNA in situ hybridization demonstrated PLP mRNA transcripts in a high proportion of oligodendrocytes in CACH patients compared to control patients, and PLP mRNA transcript signal in cells, morphologically consistent with FODCs. Normal and pathological brain control tissues did not contain FODCs. These neuropathological findings will be useful pathological identifiers of CACH, and may provide clues to the pathogenesis of this disorder. C1 Armed Forces Inst Pathol, Dept Neuropathol, Washington, DC 20306 USA. Uniformed Serv Univ Hlth Sci, Dept Anat & Cell Biol, Bethesda, MD 20814 USA. Hop St Vincent de Paul, INSERM, U342, Serv Neuropediat,AP HP, F-75674 Paris, France. Univ Texas, Med Ctr, Galveston, TX 77555 USA. Albert Einstein Coll Med, Dept Pathol, New York, NY USA. Albert Einstein Coll Med, Dept Neurosci, New York, NY USA. Univ Chicago Hosp, Dept Pathol, Chicago, IL 60637 USA. Tampa Gen Hosp, Dept Pathol, Tampa, FL 33606 USA. NINDS, Dev & Metab Neurol Branch, NIH, Bethesda, MD 20892 USA. RP Wong, K (reprint author), Armed Forces Inst Pathol, Dept Neuropathol, 16th St NW,Bldg 54, Washington, DC 20306 USA. FU NICHD NIH HHS [N01-HD-1-3138]; PHS HHS [97-N-0170, G170FN] NR 48 TC 58 Z9 59 U1 0 U2 2 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0001-6322 J9 ACTA NEUROPATHOL JI Acta Neuropathol. PD DEC PY 2000 VL 100 IS 6 BP 635 EP 646 PG 12 WC Clinical Neurology; Neurosciences; Pathology SC Neurosciences & Neurology; Pathology GA 365ZY UT WOS:000089981400007 PM 11078215 ER PT J AU Iwao, N Iwao, S Muller, DC Elahi, D Shimokata, H Andres, R AF Iwao, N Iwao, S Muller, DC Elahi, D Shimokata, H Andres, R TI A test of recently proposed BMI standards with respect to old age SO AGING-CLINICAL AND EXPERIMENTAL RESEARCH LA English DT Article; Proceedings Paper CT Annual Meeting of the Gerontological-Society-of-America CY NOV 22, 1999 CL SAN FRANCISCO, CALIFORNIA SP Gerontol Soc Amer DE age; body mass index; coronary risk factor ID DENSITY-LIPOPROTEIN CHOLESTEROL; PLASMA AB The purpose of this study teas to investigate the effects of age on the relationship between BMI and multiple coronary risk factors, and to determine whether the BMI classification by NHLBI and WHO is applicable as a predictor of coronary risk factors in older (265 years) as well as in younger (< 65 years) men and women. Effects of age on ten coronary risk factors were examined. Sex differences in the slopes of BMI on risk factors were compared between younger and older subjects in order to examine the effects of age on these relationships. The frequency of risk factor abnormality in individual BMI groups (18.5-24.9 25.029. 9, 30.0+) was examined for four age-sex groups. The significance of an age group-BMI interaction term was tested by the logistic regression model to see whether there is a significant difference in the relationship between BMI and the individual risk factor abnormalities between younger and older subjects. Older subjects had significantly higher values for most risk factors than younger subjects. The slopes of BMI on risk factors were different between younger and older subjects for fasting glucose, total, HDL- and LDL-cholesterol in men, and for diastolic blood pressure, total and LDL-cholesterol in women. The proportion of subjects with abnormal risk factor levels in each of the three BMI groups was higher in older than in younger subjects for most risk factors. There was generally a progressive worsening of the risk factor levels with increasing BMI in both age groups. There was no consistent age difference in the relationship between BMI groups and the frequency of risk factor abnormality. We conclude that, although age increases the frequency of most cardiovascular risk factor abnormalities, in general, itdoes not affect the trend of the relationship between the risk factors and the normal, overweight and obese BMI groups defined by NHLBI and WHO. Therefore, these BMI categories are applicable as predictors of risk factor levels in older as well as in younger men and women. C1 NIA, Clin Invest Lab, Metab Sect, Baltimore, MD USA. RP Andres, R (reprint author), 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 12 TC 9 Z9 9 U1 0 U2 1 PU EDITRICE KURTIS S R L PI MILAN PA VIA LUIGI ZOJA 30, 20153 MILAN, ITALY SN 0394-9532 J9 AGING-CLIN EXP RES JI Aging-Clin. Exp. Res. PD DEC PY 2000 VL 12 IS 6 BP 461 EP 469 PG 9 WC Geriatrics & Gerontology SC Geriatrics & Gerontology GA 393NE UT WOS:000166475900009 PM 11211957 ER PT J AU McDermott, DH Beecroft, MJ Kleeberger, CA Al-Sharif, EM Ollier, WER Zimmerman, PA Boatin, BA Leitman, SF Detels, R Hajeer, AH Murphy, PM AF McDermott, DH Beecroft, MJ Kleeberger, CA Al-Sharif, EM Ollier, WER Zimmerman, PA Boatin, BA Leitman, SF Detels, R Hajeer, AH Murphy, PM TI Chemokine RANTES promoter polymorphism affects risk of both HIV infection and disease progression in the Multicenter AIDS Cohort Study SO AIDS LA English DT Article DE chemokines; epidemiology; pathogenesis; risk factors; virus-cell interaction ID IMMUNODEFICIENCY-VIRUS TYPE-1; EXPOSED-UNINFECTED INDIVIDUALS; CD4(+) T-CELLS; BETA-CHEMOKINES; GENETIC RESTRICTION; PLASMODIUM-VIVAX; DELETION ALLELE; CCR5; RESISTANCE; RECEPTOR AB Objective: To examine whether polymorphism in the RANTES gene is associated with HIV disease outcome. Design: RANTES, a ligand of the major HIV co-receptor, CCR5, is known to block HIV-CCR5 interactions. Recently, two single nucleotide polymorphisms in the RANTES gene promoter region, designated -403G/A and -28C/G, have been described. Both polymorphisms can affect in-vitro promoter activity, and the RANTES -403A, -28G haplotype has been associated with a slower CD4 cell count decline rate in a Japanese cohort. Methods: We compared RANTES compound genotype frequencies between HIV-positive and exposed-uninfected participants of the Multicenter AIDS Cohort Study (MACS) and rates of progression to AIDS for MACS seroconverters. Results: We found that the two most common RANTES promoter compound genotypes, G1 (-403G/G, -28C/C) found in 67% of Caucasians, and G4 (-403G/A, -28C/C) found in 23% of Caucasians, were associated with altered risk of HIV transmission and progression, particularly in individuals who lacked the protective CCR5 mutation, CCR5 Delta 32. In this study, individuals with a G4 compound genotype were more likely to acquire HIV than individuals with a G1 compound genotype (OR 1.72, P = 0.016) and the risk increased when individuals possessing CCR5 Delta 32 were omitted from consideration (OR 2.13, P = 0.005). Among seroconverters lacking CCR5 Delta 32, those who had the G4 compound genotype progressed significantly slower to AIDS-1993 than those with the G1 compound genotype (median time to AIDS 7.6 versus 5.4 years; RH 0.65; P = 0.007). Conclusions: These data implicate the RANTES-403A allele as a risk factor for HIV transmission and as a protective factor for HIV progression. (C) 2000 Lippincott Williams & Wilkins. C1 NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. Univ Manchester, ARC, Epidemiol Unit, Manchester M13 9PT, Lancs, England. NIH, Warren Grant Magnuson Clin Ctr, Dept Transfus Med, Bethesda, MD 20892 USA. Johns Hopkins Univ, Sch Hyg & Publ Hlth, Dept Epidemiol, Baltimore, MD USA. Case Western Reserve Univ, Sch Med, Cleveland, OH USA. WHO, Onchocerciasis Control Program W Africa, Ouagadougou, Burkina Faso. Univ Calif Los Angeles, Sch Publ Hlth, Hlth Sci Ctr, Los Angeles, CA 90024 USA. RP Murphy, PM (reprint author), NIAID, Host Def Lab, NIH, Bldg 10,Room 11N113, Bethesda, MD 20892 USA. OI McDermott, David/0000-0001-6978-0867; Hajeer, Ali/0000-0003-2727-9964 FU NCRR NIH HHS [5-M01-RR-00052]; NIAID NIH HHS [UO1-AI-35042, UO1-AI-35043] NR 46 TC 162 Z9 172 U1 1 U2 6 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0269-9370 J9 AIDS JI Aids PD DEC 1 PY 2000 VL 14 IS 17 BP 2671 EP 2678 DI 10.1097/00002030-200012010-00006 PG 8 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 380TJ UT WOS:000165719300006 PM 11125885 ER PT J AU Quinn, TC Brookmeyer, R Kline, R Shepherd, M Paranjape, R Mehendale, S Gadkari, DA Bollinger, R AF Quinn, TC Brookmeyer, R Kline, R Shepherd, M Paranjape, R Mehendale, S Gadkari, DA Bollinger, R TI Feasibility of pooling sera for HIV-1 viral RNA to diagnose acute primary HIV-1 infection and estimate HIV incidence SO AIDS LA English DT Article DE HIV incidence; serumsample pooling; p24 antigen assay; reverse transcriptase-polymerase chain reaction ID HEPATITIS-C VIRUS; LIGASE CHAIN-REACTION; INCIDENCE RATES; PREVALENCE POPULATION; BLOOD DONATIONS; URINE SAMPLES; POOLED SERA; B VIRUS; IMMUNODEFICIENCY; SEROPREVALENCE AB Objective: To develop a pooling method for detection of viral RNA for diagnosis of acute HIV infection and estimation of HIV-1 incidence. Methods: Sera from 700 consecutive seronegative patients attending sexually transmitted disease clinics in Pune, India, were screened individually for p24 antigen, and pooled into seven pools of 100 for detection of HIV-1 RNA by reverse transcriptase-polymerase chain reaction. HIV-1 incidence was calculated by the traditional cohort method, the p24 antigen method, and a multistage pooling method in which RNA-positive pools were re-analyzed in smaller pools. Results: Sera from 700 individuals were grouped into seven pools of 100, of which four were positive. These four positive pools were subdivided into eight pools of 50, of which seven were positive. The seven positive pools were subdivided into 35 pools of 10, of which 10 were positive. Based on the 10 RNA-positive pools, the point estimate of HIV-1 incidence was 19.9% per year [95% confidence interval (CI), 7.3-31.8%]. Of the 700 samples analyzed for p24 antigen, eight were positive, resulting in a point estimate of incidence of 18.5%/year (8.0-36.5%). In contrast, the incidence rate based on the traditional cohort method of follow-up was lower at 9.4%/year (4.8-16.4%) due to a low follow-up rate. Testing of individual samples from the 10 RNA-positive pools identified 10 individuals with acute primary HIV-1. Conclusion: The multistage pooling method for detection of HIV-1 RNA was more sensitive than the p24 antigen method, and was five-fold less expensive than the p24 antigen assays. Pooling samples for RNA detection was effective in estimating current incidence rates with cost savings that would be practical for use in developing countries. (C) 2000 Lippincott Williams & Wilkins. C1 Johns Hopkins Univ, Div Infect Dis, Sch Med, Baltimore, MD 21205 USA. NIAID, Immunoregulat Lab, Bethesda, MD 20892 USA. Johns Hopkins Univ, Sch Hyg & Publ Hlth, Dept Biostat, Baltimore, MD USA. Natl AIDS Res Inst, Pune, Maharashtra, India. RP Quinn, TC (reprint author), Johns Hopkins Univ, Div Infect Dis, Sch Med, 720 Rutland Ave,Ross 1159, Baltimore, MD 21205 USA. FU FIC NIH HHS [D43 TW0000]; NIAID NIH HHS [R01 AI41369-01A1]; NICHD NIH HHS [HD38209] NR 30 TC 67 Z9 74 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0269-9370 J9 AIDS JI Aids PD DEC 1 PY 2000 VL 14 IS 17 BP 2751 EP 2757 DI 10.1097/00002030-200012010-00015 PG 7 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 380TJ UT WOS:000165719300015 PM 11125894 ER PT J AU Greensill, J Sheldon, JA Murthy, KK Bessonette, JS Beer, BE Schulz, TF AF Greensill, J Sheldon, JA Murthy, KK Bessonette, JS Beer, BE Schulz, TF TI A chimpanzee rhadinovirus sequence related to Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8: increased detection after HIV-1 infection in the absence of disease SO AIDS LA English DT Article DE animal models; chimpanzee; Kaposi's sarcoma; Kaposi's sarcoma-associated herpesvirus; human herpesvirus 8 ID MULTICENTRIC CASTLEMANS DISEASE; IMMUNODEFICIENCY-VIRUS TYPE-1; DNA-SEQUENCES; RETROPERITONEAL FIBROMATOSIS; PERIPHERAL-BLOOD; HUMAN-HERPESVIRUS-8; AIDS; IDENTIFICATION; PROTEIN; RHESUS AB Objective: To look for a virus related to Kaposi's sarcoma-associated herpesvirus (KSHV) or human herpesvirus 8 (HHV8) in chimpanzees and to investigate phylogenetic and biological similarities to KSHV. Methods: Peripheral blood mononuclear cell (PBMC) DNA samples from chimpanzees (Pan troglodytes troglodytes) were screened with newly designed consensus oligonucleotide primers for the DNA polymerase gene of KSHV-related gamma2-herpes-viruses (rhadinoviruses). Samples from HIV-l-infected and -uninfected chimpanzees were screened with virus-specific primers. Antibodies to KSHV structural and latent antigens were measured by immunofluorescence, enzyme-linked immunosorbent assay (ELISA) and Western blot. Results: We identified 972 base pairs (bp) of a new viral DNA polymerase sequence with 81.6% (nucleotides) and 93.2% (protein) identity to that of KSHV/HHV8. It was detected in 15/37 (41%) animals experimentally infected with HIV-1, but only in one out of 30 uninfected animals (P < 0.001). Antibodies were found by immunofluorescence to structural, but not latent, KSHV antigens in nearly all HIV-l-infected and uninfected animals. Conclusion: Like man and two other Old World primate species, chimpanzees harbour a virus closely related to KSHV/HHV8, termed Pan troglodytes rhadinovirus-1 (PtRV-1). Like KSHV, PtRV-1 is more easily detected by polymerase chain reaction (PCR) in the PBMC of HIV-l-infected than of HIV-1-uninfected individuals, suggesting increased viral load. Despite the close phylogenetic relationship and biological similarities between KSHV and PtRV-1, Kaposi's sarcoma (KS) has not been reported in HIV-1-infected chimpanzees. PtRV-1 may lack some of the pathogenic determinants of KSHV, or humans and chimpanzees may differ in how they control the infection with their respective rhadinoviruses. (C) 2000 Lippincott Williams & Wilkins. C1 Hannover Med Sch, Dept Virol, D-30625 Hannover, Germany. Univ Liverpool, Dept Med Microbiol & Genitourinary Med, Mol Virol Grp, Liverpool L69 3BX, Merseyside, England. SW Fdn Med Res, Dept Virol & Immunol, San Antonio, TX USA. NIAID, Mol Microbiol Lab, NIH, Rockville, MD USA. RP Schulz, TF (reprint author), Hannover Med Sch, Dept Virol, Carl Neuberg Str 1, D-30625 Hannover, Germany. NR 41 TC 24 Z9 26 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0269-9370 J9 AIDS JI Aids PD DEC 1 PY 2000 VL 14 IS 17 BP F129 EP F135 DI 10.1097/00002030-200012010-00001 PG 7 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 380TJ UT WOS:000165719300001 PM 11125908 ER PT J AU Rogers, AS Lindsey, JC Futterman, DC Zimmer, B Abdalian, SE D'Angelo, LJ AF Rogers, AS Lindsey, JC Futterman, DC Zimmer, B Abdalian, SE D'Angelo, LJ CA Pediat AIDS Clin Trial Group Proto TI Serologic examination of hepatitis B infection and immunization in HIV-positive youth and associated risks SO AIDS PATIENT CARE AND STDS LA English DT Article ID UNITED-STATES; VIRUS-INFECTION; ADOLESCENTS; HEALTH AB This seroprevalence report examines serologic evidence of hepatitis B immunization or infection and associated demographic/behavioral factors in adolescent (aged 12-20) subjects enrolled in a nontherapeutic clinical trial at 43 Pediatric AIDS Clinical Trials Group (PACTG) clinical centers. Subjects (n = 94) infected with the human immunodeficiency virus (HIV) through sexual activity were categorized as hepatitis B virus (HBV)-immunized, HBV-infected, or nonimmune by hepatitis B serology performed on specimens collected within the subject's first 48 weeks on study (1993-1995). Sixteen percent of the 94 serologically classified subjects were immunized; 19% HBV-infected; 65% nonimmune. Of the three risk factor scores examined (sociodemographic, sexual, and substance abuse), substance use alone demonstrated a significant difference among groups (despite virtually no reported injecting drug behavior), with the sexual risk score exhibiting marginally significant differences. Logistic regression analysis (restricted to nonimmunized subjects) showed that male-male sexual activity raised the odds of HBV infection by a factor of 5.14 (95% confidence interval [CI]: 1.45-18.23) relative to heterosexual activity; and that for every one point increase on the substance abuse risk scale the odds of infection increased 5% (95% CI: 0.99-1.10). The HBV infection rate in PACTG 220 HIV-positive females is twice United States population-based rates; the rate in PACTG 220 HIV-positive males is nearly seven times higher. Past immunization efforts in this population appear to have been based on sexual activity volume without regard to injecting-drug use in sex partners. C1 NICHD, Pediat Adolescent & Maternal AIDS Branch, CRMC, NIH, Bethesda, MD 20892 USA. Harvard Univ, Sch Publ Hlth, Boston, MA 02115 USA. Albert Einstein Coll Med, New York, NY USA. Frontier Sci & Technol Res Fdn Inc, Amherst, NY USA. Tulane Univ, Sch Med, New Orleans, LA 70112 USA. Childrens Natl Med Ctr, Washington, DC 20010 USA. RP Rogers, AS (reprint author), NICHD, Pediat Adolescent & Maternal AIDS Branch, CRMC, NIH, Execut Bldg Room 4B11,6100 Execut Blvd MSC 7510, Bethesda, MD 20892 USA. FU NIAID NIH HHS [1UO1-AI-41110]; NICHD NIH HHS [N01-HD-3-3162] NR 11 TC 12 Z9 13 U1 0 U2 0 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1087-2914 J9 AIDS PATIENT CARE ST JI Aids Patient Care STDS PD DEC PY 2000 VL 14 IS 12 BP 651 EP 657 DI 10.1089/10872910050206577 PG 7 WC Public, Environmental & Occupational Health; Infectious Diseases SC Public, Environmental & Occupational Health; Infectious Diseases GA 383UD UT WOS:000165900500006 PM 11119432 ER PT J AU Padow, M Lai, LL Fisher, RJ Zhou, YC Wu, XY Kappes, JC Towler, EM AF Padow, M Lai, LL Fisher, RJ Zhou, YC Wu, XY Kappes, JC Towler, EM TI Analysis of human immunodeficiency virus type 1 containing HERV-K protease SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article ID HUMAN-ENDOGENOUS-RETROVIRUS; PRECURSOR PROTEIN; PARTICLES; VPX; IDENTIFICATION; SPECIFICITY; FUSION; GENE; LOOP AB The human endogenous retrovirus, type K (HERV-K) represents the most biologically active form of known retroelements present in the human genome. Several HERV-K genomes have transcriptionally active open reading frames and encode their own protease (PR). The HERV-K PR has been shown to authentically cleave human immunodeficiency virus type 1 (HIV-1) matrix-capsid peptide in the presence of HIV-1 PR inhibitors. This raised the possibility that HERV-K PR could complement HIV-1 PR function in HIV-1-infected individuals. To investigate this possibility, we fused the HIV-1 vpr gene to the HERV-K PR gene (vpr-PR). The vpr-PR expression plasmid and a PR-defective HIV-1 clone were cotransfected into 293T cells. Progeny virions were assayed for processing of the HIV-1 polyproteins by Western blot and for changes in infectivity. HERV-K PR fused to Vpr was incorporated into HIV-1 virions at a high concentration and cleaved the Gag and Pol precursor proteins. However, neither Gag nor Pol polyproteins were correctly processed. Moreover, the HERV-K PR did not restore virus infectivity. While these results do not exclude the possibility that the HERV-K PR could complement an HIV-1 PR whose function is impaired due to drugs or drug-resistant mutations, they clearly demonstrate that the HERV-K PR cannot substitute for the function of the wild-type HIV-1 PR. C1 Univ Alabama, Dept Microbiol, Birmingham, AL 35294 USA. Univ Alabama, Dept Med, Birmingham, AL 35294 USA. NCI, Frederick Canc Res & Dev Ctr, Frederick, MD 21701 USA. Birmingham Vet Affairs, Med Ctr Res Serv, Birmingham, AL 35233 USA. RP Towler, EM (reprint author), Bayer Corp, Dept Prot Biosci, 400 Morgan Lane, W Haven, CT 06516 USA. RI Fisher, Robert/B-1431-2009 FU NCI NIH HHS [CA 73470, N01-CO-56000]; NIGMS NIH HHS [T32 GM0811] NR 23 TC 12 Z9 12 U1 0 U2 4 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 0889-2229 J9 AIDS RES HUM RETROV JI Aids Res. Hum. Retrovir. PD DEC PY 2000 VL 16 IS 18 BP 1973 EP 1980 DI 10.1089/088922200750054701 PG 8 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 389DM UT WOS:000166223400006 PM 11153080 ER PT J AU Mahieux, R Chappey, C Meertens, L Mauclere, P Lewis, J Gessain, A AF Mahieux, R Chappey, C Meertens, L Mauclere, P Lewis, J Gessain, A TI Molecular characterization and phylogenetic analyses of a new simian T cell lymphotropic virus type 1 in a wild-caught African baboon (Papio anubis) with an indeterminate STLV type 2-like serology SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article ID INTERSPECIES TRANSMISSION; PAN-PANISCUS; POPULATIONS; STRAINS; SUBTYPE AB STLV-1 viruses are closely related to HTLV-1 and infect many African monkey species. Seroreactivities of monkeys infected by STLV-1 are nearly identical to those of HTLV-1-infected individuals. In some cases, STLV-1 are, sequence-wise, indistinguishable from HTLV-1, and cannot be separated from them on the basis of phylogenetic analyses. HTLV-2-related simian viruses have been rarely reported. Such STLV-2 viruses, present in African bonobo (Pan paniscus), possess a genomic organization related to but different from all known HTLV-2 subtypes. We report here the molecular characterization and the subtyping of a new STLV-1 in a wild-caught baboon (Papio anubis) whose serum exhibited an indeterminate STLV-2-like serology (p24, GD21, MTA-1 with no p19). In the env and LTR regions, this virus is phylogenetically related to the large African STLV-1 group, but does not cluster with any STLV-1 baboon sequence. The complete p19 sequence reveals amino acid changes at critical positions. This is the first report of an African STLV-1 virus leading to an STLV-2-like serological profile in its host. C1 Inst Pasteur, Unite Oncol Virale, F-75724 Paris 15, France. Natl Ctr Biotechnol Informat, NIH, Bethesda, MD 20892 USA. Ctr Pasteur Cameroun, Yaounde, Cameroon. Int Zoo Vet Grp, Keighley BD21 1AG, W Yorkshire, England. RP Mahieux, R (reprint author), Inst Pasteur, Unite Oncol Virale, 28 Rue Docteur Roux, F-75724 Paris 15, France. RI Meertens, Laurent/E-8043-2017 NR 17 TC 13 Z9 13 U1 0 U2 0 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 0889-2229 J9 AIDS RES HUM RETROV JI Aids Res. Hum. Retrovir. PD DEC PY 2000 VL 16 IS 18 BP 2043 EP 2048 DI 10.1089/088922200750054774 PG 6 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 389DM UT WOS:000166223400013 PM 11153087 ER PT J AU Newlin, DB Miles, DR van den Bree, MBM Gupman, AE Pickens, RW AF Newlin, DB Miles, DR van den Bree, MBM Gupman, AE Pickens, RW TI Environmental transmission of DSM-IV substance use disorders in adoptive and step families SO ALCOHOLISM-CLINICAL AND EXPERIMENTAL RESEARCH LA English DT Article DE alcoholism; drug abuse and drug dependence; environmental; genetic; adoption ID ALCOHOL EPIDEMIOLOGIC SURVEY; POPULATION-BASED SAMPLE; DRUG-USE; FEMALE TWINS; GENETIC INFLUENCES; OPIOID ADDICTS; UNITED-STATES; DEPENDENCE; ABUSE; PSYCHOPATHOLOGY AB Background: One factor contributing to the 3- to 5-fold increase in risk for substance use disorders (SUDs) among children of alcoholics may be the rearing environment. These influences may include availability of substances, modeling of SUDs, inadequate parenting, or other factors. The contribution of parental environmental influences on offspring with SUDs may be estimated independently of genetic influences through assessment of adoptees raised by nonbiological parents. Methods: Relative risk of SUDs was assessed in adult adoptees (N = 442) of alcoholic and nonalcoholic adoptive parents as well as in stepchildren (N = 1859) with alcoholic or nonalcoholic stepfathers who participated in the community-based National Longitudinal Alcohol Epidemiologic Survey (NLAES). Results: Rearing by an alcoholic adoptive mother was associated with increased DSM-IV alcohol abuse. Rearing by an alcoholic adoptive father was predictive of adoptees' illicit drug use, as well as DSM-IV drug dependence. Rearing by an alcoholic stepfather was predictive of stepchild DSM-IV alcohol abuse, illicit drug use, and drug dependence, whereas an alcoholic stepmother was associated with increased illicit drug use in the stepchild. Alcoholism in adoptive parents or step parents did not increase risk for offspring DSM-IV alcohol dependence. In both adoptive and biological families, there was a subadditive interaction of mother by father alcoholism such that the rate of substance abuse when both parents were alcoholic was less than that expected based on the additive effects of each alcoholic parent. Conclusions: Rearing by an alcoholic parent had a greater influence on alcohol abuse by offspring than on alcohol dependence. The increased risk of proband illicit drug use and drug dependence associated with paternal alcoholism suggested nonspecificity of environmental transmission. Both maternal and paternal cultural transmission effects influenced offspring SUDs. C1 NIDA, Intramural Res Program, Baltimore, MD 21224 USA. Virginia Commonwealth Univ, Richmond, VA USA. Biognosis US Inc, Gaithersburg, MD USA. RP Newlin, DB (reprint author), NIDA, Intramural Res Program, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. RI turton, miranda/F-4682-2011 NR 61 TC 24 Z9 24 U1 1 U2 3 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0145-6008 J9 ALCOHOL CLIN EXP RES JI Alcoholism (NY) PD DEC PY 2000 VL 24 IS 12 BP 1785 EP 1794 DI 10.1111/j.1530-0277.2000.tb01982.x PG 10 WC Substance Abuse SC Substance Abuse GA 383YB UT WOS:000165911300007 PM 11141037 ER PT J AU Dawson, DA AF Dawson, DA TI US low-risk drinking guidelines: An examination of four alternatives SO ALCOHOLISM-CLINICAL AND EXPERIMENTAL RESEARCH LA English DT Article ID ALCOHOL; HEALTH AB Background: This study compared four sets of US low-risk drinking guidelines (two interpretations of the US Dietary Guidelines and two variations of the NIAAA physicians' guidelines) in terms of adherence and how well they predicted five different alcohol-related outcomes. Methods: Using data from a nationally representative sample of 17,542 US adults 21 years of age and over who drank 12 or more drinks in the past year, this study assessed the sensitivity, specificity, overall accuracy, positive and negative predictive values, and odds ratios of the various drinking guidelines (specifically, of having exceeded them with different degrees of frequency) as predictors of alcohol dependence, impaired driving, liver disease, peptic ulcer, and hypertension. Results: The proportions of past-year regular drinkers exceeding the four sets of guidelines varied from 20.9%, whose average intake exceeded the weekly Limits, to between 21.0% and 42.7% who exceeded the daily guidelines at least once a week, and to between 69.2% and 94.2% who ever exceeded the daily limits in the year preceding the interview. Sensitivity and odds ratios were highest for the ever exceeding the Dietary Guidelines daily limits, intermediate for ever exceeding the two variations based on the NIAAA physicians' guidelines, and lowest far exceeding the Dietary Guidelines interpreted as weekly limits. The opposite pattern was observed for specificity and overall predictive accuracy. When frequently exceeding the daily limits was considered, their sensitivity declined but their specificity and positive predictive value increased. Conclusions: If sensitivity and specificity are deemed equally important, the NIAAA physicians' guidelines incorporating both daily and weekly limits seem to do the best job of balancing these dimensions in the prediction of a variety of alcohol-related outcomes. C1 NIAAA, Div Biometry & Epidemiol, NIH, Bethesda, MD 20892 USA. RP Dawson, DA (reprint author), NIAAA, Div Biometry & Epidemiol, NIH, Willco Bldg,Suite 514,6000 Execut Bldg,MSC 7003, Bethesda, MD 20892 USA. NR 32 TC 58 Z9 58 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0145-6008 J9 ALCOHOL CLIN EXP RES JI Alcoholism (NY) PD DEC PY 2000 VL 24 IS 12 BP 1820 EP 1829 DI 10.1111/j.1530-0277.2000.tb01986.x PG 10 WC Substance Abuse SC Substance Abuse GA 383YB UT WOS:000165911300011 PM 11141041 ER PT J AU Semba, RD Garrett, E Johnson, BA Guralnik, JM Fried, LP AF Semba, RD Garrett, E Johnson, BA Guralnik, JM Fried, LP TI Vitamin D deficiency among older women with and without disability SO AMERICAN JOURNAL OF CLINICAL NUTRITION LA English DT Article DE vitamin D deficiency; calcium; bone fractures; disability; women; aging; Baltimore ID POSTMENOPAUSAL WOMEN; PLASMA 25-HYDROXYVITAMIN-D; D SUPPLEMENTATION; ELDERLY WOMEN; BONE-DENSITY; HIP FRACTURE; CALCIUM; POPULATION; SERUM; ASSAY AB Background: Vitamin D deficiency is associated with bone loss and bone fractures, and the identification of vulnerable populations is important to clinical practice and public health. Objective: The objectives of this study were to determine the prevalence of vitamin D deficiency and to examine associated risk factors for vitamin D deficiency in older women. Design: We measured serum concentrations of 25-hydroxyvitamin D [25(OH)D], 1,25-dihydroxyvitamin D [1,25(OH)(2)D], intact parathyroid hormone (PTH), osteocalcin, and ionized calcium in women aged greater than or equal to 65 y who were participating in the Women's Health and Aging Study I, an observational study of women representing the approximately one-third most disabled women living in the community, and women aged 70-80 y who were participating in the Women's Health and Aging Study I, an observational study of women among the two-thirds Least disabled women living in the community in Baltimore. Results: The women were classified into 3 domains of physical disability. Among 371 women with 0 or 1 domain of disability and 682 women with greater than or equal to 2 domains of disability, 6.2% and 12.6%, respectively, had vitamin D deficiency [serum concentrations of 25(OH)D < 25 nmol/L]. In univariate analyses, risk factors for vitamin D deficiency included increasing age, black race, low educational level, high body mass index, high triceps skinfold thickness, increasing level of disability, winter season, and elevated creatinine concentration. In multivariate models, black race had a strong association with vitamin D deficiency when other risk factors were adjusted for. Conclusions: Vitamin D deficiency, a preventable disorder, is a common and important public health problem for older disabled women living in the community; black women are at higher risk than are white women. C1 Johns Hopkins Med Inst, Baltimore, MD 21205 USA. NIA, Epidemiol Demog & Biometry Program, Bethesda, MD 20892 USA. RP Semba, RD (reprint author), 550 N Broadway,Suite 700, Baltimore, MD 21205 USA. FU NIA NIH HHS [N01AG122112, R01AG11703]; NIAID NIH HHS [R01AI41956] NR 42 TC 60 Z9 65 U1 1 U2 6 PU AMER SOC CLINICAL NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, SUBSCRIPTIONS, RM L-3300, BETHESDA, MD 20814-3998 USA SN 0002-9165 J9 AM J CLIN NUTR JI Am. J. Clin. Nutr. PD DEC PY 2000 VL 72 IS 6 BP 1529 EP 1534 PG 6 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 377WJ UT WOS:000165549000020 PM 11101482 ER PT J AU Lei, JY Wang, Y Jaffe, ES Turner, ML Raffeld, M Sorbara, L Morris, J Holland, SM Duray, PH AF Lei, JY Wang, Y Jaffe, ES Turner, ML Raffeld, M Sorbara, L Morris, J Holland, SM Duray, PH TI Microcystic adnexal carcinoma associated with primary immunodeficiency, recurrent diffuse herpes simplex virus infection, and cutaneous T-cell lymphoma SO AMERICAN JOURNAL OF DERMATOPATHOLOGY LA English DT Article DE antecubital fossa; herpes virus; microcystic adnexal carcinoma; peripheral T-cell lymphoma ID DIFFERENTIATION; SERIES AB Cutaneous microcystic adnexal carcinoma (MAC) is a rare and poorly understood tumor that predominantly occurs in the head and neck. MAC usually affects people in their fourth and fifth decades. Some patients have had a history of radiation. We present a case of MAC occurring in the left antecubital fossa of an 18-year-old white woman with an unusual immunodeficiency syndrome. The patient also developed a squamous cell carcinoma, a cutaneous T-cell malignancy, and a perigastric leiomyoma. A congenital infection of herpes simplex virus (HSV) persisted throughout her life. The association of HSV infection with MAC and squamous cell carcinoma and that of peripheral T-cell lymphoma with Epstein-Barr virus is discussed in relation to her immunodeficiency. C1 NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. NCI, Dermatol Branch, NIH, Bethesda, MD 20892 USA. NCI, Metab Branch, NIH, Bethesda, MD 20892 USA. NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. RP Duray, PH (reprint author), NCI, Pathol Lab, NIH, 10-2N212,10 Ctr Dr, Bethesda, MD 20892 USA. NR 23 TC 19 Z9 21 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0193-1091 J9 AM J DERMATOPATH JI Am. J. Dermatopathol. PD DEC PY 2000 VL 22 IS 6 BP 524 EP 529 DI 10.1097/00000372-200012000-00008 PG 6 WC Dermatology SC Dermatology GA 384FW UT WOS:000165933000008 PM 11190445 ER PT J AU Hadley, EC Rossi, WK Albert, S Bailey-Wilson, J Baron, J Cawthon, R Christian, JC Corder, EH Franceschi, C Kestenbaum, B Kruglyak, L Lauderdale, DS Lubitz, J Martin, GM McClearn, GE McGue, M Miles, T Mineau, G Ouellett, G Pedersen, NL Preston, SH Page, WF Province, M Schachter, F Schork, NJ Vaupel, JW Vijg, J Wallace, R Wang, E Wijsman, EM AF Hadley, EC Rossi, WK Albert, S Bailey-Wilson, J Baron, J Cawthon, R Christian, JC Corder, EH Franceschi, C Kestenbaum, B Kruglyak, L Lauderdale, DS Lubitz, J Martin, GM McClearn, GE McGue, M Miles, T Mineau, G Ouellett, G Pedersen, NL Preston, SH Page, WF Province, M Schachter, F Schork, NJ Vaupel, JW Vijg, J Wallace, R Wang, E Wijsman, EM CA NIA Aging Genetic Epidemiology Wor TI Genetic epidemiologic studies on age-specified traits SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Editorial Material DE aging; disease; mortality; survival ID PHENOTYPIC COMBINATION A1B8CW7DR3; ALZHEIMERS-DISEASE; HEALTH EXPECTANCY; HUMAN LONGEVITY; WOMEN TWINS; OLDER; RISK; ASSOCIATION; IDENTIFICATION; HERITABILITY AB This commentary calls attention to the value of combining genetic and epidemiologic methods in studies to understand the determinants of two crucial aspects of aging: ages at which certain outcomes (e.g., disease, mortality) occur and rates of change with age of individual's characteristics (e.g., physiologic functions, disease risk factors). Inclusion of age in the specification of traits in genetic epidemiologic studies could lead to improved strategies to increase healthy life expectancy and evaluate individuals' risk for age-related morbidity. Special issues that make genetic epidemiologic approaches important for studies of age-specified phenomena as well as opportunities and challenges for such studies are discussed, including study designs, sampling frames, databases, analytic tools, and related methodological issues. This commentary is based on a report prepared by the Aging and Genetic Epidemiology Working Group, convened by the National Institute on Aging to review opportunities for research on the genetic epidemiology of aging-related outcomes. The report, which contains more extensive discussion, literature review, and references, is available on the World Wide Web at http://www.nih.gov/nia/conferences/GenetReport111199.htm. Am J Epidemiol 2000;152;1003-8. C1 NIA, Geriatr Program, Bethesda, MD 20892 USA. Columbia Univ, Gertrude H Sergievsky Ctr, New York, NY 10027 USA. NHGRI, Baltimore, MD USA. Dartmouth Coll, Hitchcock Med Ctr, Dartmouth Med Sch, Dept Med, Hanover, NH 03756 USA. Dartmouth Coll, Hitchcock Med Ctr, Dartmouth Med Sch, Dept Community & Family Practice, Hanover, NH 03756 USA. Univ Utah, Dept Human Genet, Salt Lake City, UT USA. Indiana Univ, Dept Med & Mol Genet, Indianapolis, IN 46204 USA. Odense Univ, Ctr Genet Epidemiol, Odense, Denmark. Dept Biomed Sci, Sect Gen Pathol, Modena, Italy. US Social Secur Adm, Baltimore, MD USA. MIT, Whitehead Inst Biomed Res, Ctr Genome Res, Cambridge, MA USA. Univ Chicago, Dept Hlth Studies, Chicago, IL 60637 USA. US Hlth Care Financing Adm, Off Strateg Planning, Baltimore, MD 21207 USA. Univ Washington, Dept Pathol, Seattle, WA 98195 USA. Penn State Univ, Ctr Dev & Hlth Genet, University Pk, PA 16802 USA. Univ Minnesota, Dept Psychol, Minneapolis, MN 55455 USA. Univ Texas, Hlth Sci Ctr, Dept Family Practice, San Antonio, TX USA. Univ Utah, Dept Oncol Sci, Div Publ Hlth Sci, Salt Lake City, UT USA. Algene Biotechnol, Genet Epidemiol Lab, Montreal, PQ, Canada. Karolinska Inst, Inst Environm Med, S-10401 Stockholm, Sweden. Univ Penn, Populat Study Ctr, Philadelphia, PA 19104 USA. Natl Acad Sci, Inst Med, Med Follow Up Agcy, NAS NRC Twin Registry, Washington, DC 20418 USA. Washington Univ, Med Ctr, Sch Med, Div Biostat, St Louis, MO USA. Univ Leonardo da Vinci, ISMCM, CESTI, Paris, France. Metrohlth Med Ctr, Dept Epidemiol & Biostat, Cleveland, OH USA. Max Planck Inst Demog Res, Rostock, Germany. Univ Texas, Hlth Sci Ctr, Dept Physiol, San Antonio, TX 78284 USA. Univ Iowa, Dept Prevent Med, Iowa City, IA 52242 USA. Sir Mortimer B Davis Jewish Hosp, Lady Davis Inst Med Res, Bloomfield Ctr Res Aging, Montreal, PQ H3T 1E2, Canada. Univ Washington, Dept Med, Div Med Genet, Seattle, WA 98195 USA. Univ Washington, Dept Biostat, Seattle, WA 98195 USA. RP Hadley, EC (reprint author), NIA, Geriatr Program, 7201 Wisconsin Ave,Gateway Bldg,Suite 3E327, Bethesda, MD 20892 USA. OI Bailey-Wilson, Joan/0000-0002-9153-2920; Miles, Toni/0000-0003-0823-2319 NR 38 TC 19 Z9 20 U1 1 U2 4 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD DEC 1 PY 2000 VL 152 IS 11 BP 1003 EP 1008 PG 6 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 378DA UT WOS:000165568700001 ER PT J AU Ruhl, CE Everhart, JE AF Ruhl, CE Everhart, JE TI Association of coffee consumption with gallbladder disease SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Article DE cholelithiasis; coffee; gallbladder diseases; nutrition surveys; risk factors ID GALLSTONE DISEASE; PHYSICAL-ACTIVITY; RISK-FACTORS; SMOKING; PREVALENCE; WEIGHT AB Coffee consumption was recently shown to protect against symptomatic gallbladder disease in men. The authors examined the relation of ultrasound-documented gallbladder disease with coffee drinking in 13,938 adult participants in the Third National Health and Nutrition Examination Survey, 1988-1994. The prevalence of total gallbladder disease was unrelated to coffee consumption in either men or women. However, among women a decreased prevalence of previously diagnosed gallbladder disease was found with increasing coffee drinking (p = 0.027). These findings do not support a protective effect of coffee consumption on total gallbladder disease, although coffee may decrease the risk of symptomatic gallstones in women. C1 Social & Sci Syst Inc, Bethesda, MD 20814 USA. NIDDKD, NIH, Bethesda, MD 20892 USA. RP Ruhl, CE (reprint author), Social & Sci Syst Inc, 7101 Wisconsin Ave,Suite 1300, Bethesda, MD 20814 USA. FU NIDDK NIH HHS [N01-DK-6-2220] NR 14 TC 20 Z9 21 U1 1 U2 1 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD DEC 1 PY 2000 VL 152 IS 11 BP 1034 EP 1038 DI 10.1093/aje/152.11.1034 PG 5 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 378DA UT WOS:000165568700006 PM 11117612 ER PT J AU El-Serag, HB Everhart, JE AF El-Serag, HB Everhart, JE TI Improved survival after variceal hemorrhage over an 11-year period in the Department of Veterans Affairs SO AMERICAN JOURNAL OF GASTROENTEROLOGY LA English DT Article ID PORTAL-HYPERTENSION; RECORDS; LIVER; FILES AB OBJECTIVES: Over the past two decades, several modalities have become widely used in the management of esophageal variceal hemorrhage. The effectiveness of these measures on the outcome of patients with this type of hemorrhage remains unknown. METHODS: Using the Department of Veterans Affairs (VA) Patient Treatment File, we identified two cohorts of patients diagnosed with an initial variceal hemorrhage: an early cohort during 1981-1982 (1339 patients), and a late cohort during 1988-1991 (3636 patients). Each cohort was followed for 6 yr for rebleeding and death. Analyses were performed with proportional hazards survival analysis controlling for confounding factors. RESULTS: On presentation, patients in the late cohort were older (57 yr vs 55 yr, p < 0.0001) and had more ascites (25% vs 13%, p < 0.0001), more peritonitis (4% vs 2%, p < 0.0001), and more encephalopathy (14% vs 9%, p = 0.0003). The late cohort experienced a significant decline in mortality at 30 days (20.8% vs 29.6%, p = 0.0001) and at 6 yr (69.7% vs 74.5%, p = 0.0001). This improvement was accentuated in multivariate survival analysis when controlling for the more severe illness in the late cohort. For patients who survived the first 30 days, no significant difference in 6-yr mortality was found on univariate analysis between the early cohort (63.7%) and late cohort (61.8%) (p = 0.25), but survival was slightly better in the late cohort on multivariate analysis (p = 0.01). In the late cohort, patients with sclerotherapy during the initial hospitalization had better 30-day (17%) and 6-yr mortality (68%) than did the rest of the late cohort. CONCLUSIONS: Between the years 1981-1982 and 1988-1991, improvements in long-term survival after an initial episode of esophageal variceal hemorrhage resulted primarily from better short-term mortality. Sclerotherapy offers a partial explanation for improved survival. (C) 2000 by Am. Cell. of Gastroenterology. C1 Houston Vet Affairs Med Ctr, Gastroenterol Sect, Houston, TX 77030 USA. Houston Vet Affairs Med Ctr, Sect Hlth Serv Res, Houston, TX 77030 USA. Baylor Coll Med, Houston, TX 77030 USA. NIDDK, Bethesda, MD USA. RP El-Serag, HB (reprint author), Houston Vet Affairs Med Ctr, Gastroenterol Sect, 152,2002 Holcombe Blvd, Houston, TX 77030 USA. NR 30 TC 112 Z9 117 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0002-9270 J9 AM J GASTROENTEROL JI Am. J. Gastroenterol. PD DEC PY 2000 VL 95 IS 12 BP 3566 EP 3573 PG 8 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 383GC UT WOS:000165873900035 PM 11151893 ER PT J AU Bellus, GA Spector, EB Speiser, PW Weaver, CA Garber, AT Bryke, CR Israel, J Rosengren, SS Webster, MK Donoghue, DJ Francomano, CA AF Bellus, GA Spector, EB Speiser, PW Weaver, CA Garber, AT Bryke, CR Israel, J Rosengren, SS Webster, MK Donoghue, DJ Francomano, CA TI Distinct missense mutations of the FCFR3 Lys650 codon modulate receptor kinase activation and the severity of the skeletal dysplasia phenotype SO AMERICAN JOURNAL OF HUMAN GENETICS LA English DT Article ID GROWTH-FACTOR RECEPTOR-3; THANATOPHORIC DYSPLASIA; ACANTHOSIS NIGRICANS; GENETIC-HETEROGENEITY; DEVELOPMENTAL DELAY; HYPOCHONDROPLASIA; ACHONDROPLASIA; TRANSMEMBRANE; DOMAIN; FGFR3 AB The fibroblast growth factor-receptor 3 (FGFR3) Lys650 codon is located within a critical region of the tyrosine kinase-domain activation loop, Two missense mutations in this codon are known to result in strong constitutive activation of the FGFR3 tyrosine kinase and cause three different skeletal dysplasia syndromes-thanatophoric dysplasia type II (TD2) (A1948G [Lys650Glu]) and SADDAN (severe achondroplasia with developmental delay and acanthosis nigricans) syndrome and thanatophoric dysplasia type I (TD1) (both due to A1949T [Lys650Met]). Other mutations within the FGFR3 tyrosine kinase domain (e.g., C1620A or C1620G [both resulting in Asn540Lys]) are known to cause hypochondroplasia, a relatively common but milder skeletal dysplasia. In 90 individuals with suspected clinical diagnoses of hypochondroplasia who do not have Asn540Lys mutations, we screened for mutations, in FGFR3 exon 15, that would disrupt a unique BbsI restriction site that includes the Lys650 codon. We report here the discovery of three novel mutations (G1950T and G1950C [both resulting in Lys650Asn] and A1948C [Lys650Gln]) occurring in six individuals from five families. Several physical and radiological features of these individuals were significantly milder than those in individuals with the Asn540Lys mutations. The Lys650Asn/Gln mutations result in constitutive activation of the FGFR3 tyrosine kinase but to a lesser degree than that observed with the Lys540Glu and Lys650Met mutations. These results demonstrate that different amino acid substitutions at the FGFR3 Lys650 codon can result in several different skeletal dysplasia phenotypes. C1 Univ Colorado, Sch Med, Dept Dermatol, Denver, CO 80262 USA. Univ Colorado, Sch Med, Dept Pediat, Denver, CO 80262 USA. Univ Colorado, Sch Med, Ctr Med Genet, Denver, CO 80262 USA. N Shore Univ Hosp, Dept Pediat, NYU, Sch Med, Manhasset, NY 11030 USA. Carle Clin Assoc, Div Med Genet, Dept Pediat, Urbana, IL USA. Comprehens Genet Serv, Milwaukee, WI USA. Univ Connecticut, Sch Med, Dept Pediat, Farmington, CT 06032 USA. Univ Calif San Diego, Ctr Mol Genet, Dept Chem & Biochem, La Jolla, CA 92093 USA. NHGRI, Med Genet Branch, NIH, Bethesda, MD 20892 USA. RP Bellus, GA (reprint author), Univ Colorado, Sch Med, Dept Dermatol, B-153,4200 E 9th Ave, Denver, CO 80262 USA. FU NIAMS NIH HHS [K08 AR002075, 1K08AR002075]; NIDCR NIH HHS [R01-DE12581] NR 41 TC 95 Z9 101 U1 1 U2 4 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0002-9297 J9 AM J HUM GENET JI Am. J. Hum. Genet. PD DEC PY 2000 VL 67 IS 6 BP 1411 EP 1421 DI 10.1086/316892 PG 11 WC Genetics & Heredity SC Genetics & Heredity GA 378WE UT WOS:000165607600006 PM 11055896 ER PT J AU Hill, DA Weiss, NS Beresford, SAA Voigt, LF Daling, JR Stanford, JL Self, S AF Hill, DA Weiss, NS Beresford, SAA Voigt, LF Daling, JR Stanford, JL Self, S TI Continuous combined hormone replacement therapy and risk of endometrial cancer SO AMERICAN JOURNAL OF OBSTETRICS AND GYNECOLOGY LA English DT Article DE endometrial neoplasms; estrogen replacement therapy; estrogens; menopause; progestational hormones ID POSTMENOPAUSAL WOMEN; ESTROGEN REPLACEMENT; MEDROXYPROGESTERONE ACETATE; CONJUGATED ESTROGENS; EXOGENOUS ESTROGENS; PROGESTINS; BIOCHEMISTRY; INFORMATION; FRACTURES; SURVIVAL AB OBJECTIVE: Postmenopausal women who receive sequential hormone replacement therapy with estrogen Combined with progestogen for 10 to 24 dime for a prolonged period may have an elevated endometrial cancer risk relative to those who have never received hormone replacement therapy. We investigated whether daily use of estrogen and progestogen (continuous combined hormone replacement therapy) could diminish any excess endometrial cancer risk. STUDY DESIGN: A population-based study in Washington State obtained interview data from 969 women aged 45 to 74 years with endometrial cancer diagnosed during 1985 through 1991 or 1994 through 1995 and from 1325 age-matched control subjects selected primarily by random digit dialing. Women who had received only continuous combined hormone replacement therapy were compared with women who had only received another hormone replacement therapy regimen or who had never received hormone replacement-therapy RESULTS: The risk of endometrial cancer among users of continuous combined hormone replacement therapy (n = 9 case patients, n = 33 control subjects) relative to women who had never received hormone replacement therapy was 0.6 (95% confidence interval, 0.3-1.3); the risk relative to women who received hormone replacement that included progestogen for 10 to 24 dime was 0.4 (95% confidence interval, 0.2-1.1). Most Continuous combined hormone replacement therapy use was short-term (<72 months) or recent tin the previous; 24 months). CONCLUSION: Women who had received continuous combined hormone replacement therapy for several years did not appear to be at any increased risk for endometrial cancer relative to women who had never received hormone replacement therapy and may in fact be at decreased risk for endometrial cancer. C1 Univ Washington, Dept Epidemiol, Seattle, WA 98195 USA. Univ Washington, Dept Biostat, Seattle, WA 98195 USA. Fred Hutchinson Canc Res Ctr, Div Publ Hlth Sci, Seattle, WA 98104 USA. RP Hill, DA (reprint author), NCI, Div Canc Epidemiol & Genet, 6120 Execut Blvd,MSC 7238, Bethesda, MD 20892 USA. FU NCI NIH HHS [N01-CN-05230, R01-CA-39779, R35-CA-39779] NR 26 TC 59 Z9 60 U1 1 U2 3 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0002-9378 J9 AM J OBSTET GYNECOL JI Am. J. Obstet. Gynecol. PD DEC PY 2000 VL 183 IS 6 BP 1456 EP 1461 DI 10.1067/mob.2000.108081 PG 6 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 384NP UT WOS:000165952000032 PM 11120510 ER PT J AU Goepfert, AR Goldenberg, RL Mercer, B Iams, J Meis, P Moawad, A Thom, E VanDorsten, JP Caritis, SN Thurnau, G Miodovnik, M Dombrowski, M Roberts, JM McNellis, D AF Goepfert, AR Goldenberg, RL Mercer, B Iams, J Meis, P Moawad, A Thom, E VanDorsten, JP Caritis, SN Thurnau, G Miodovnik, M Dombrowski, M Roberts, JM McNellis, D CA Natl Inst Child Hlth Human Dev Mat TI The Preterm Prediction Study: Quantitative fetal fibronectin values and the prediction of spontaneous preterm birth SO AMERICAN JOURNAL OF OBSTETRICS AND GYNECOLOGY LA English DT Article DE preterm birth; fibronectin ID DELIVERY AB OBJECTIVE: A cervicovaginal fetal fibronectin value of greater than or equal to 50 ng/mL has been used to define women at risk of having a preterm birth. We evaluated the relationship between quantitative fetal fibronectin values and spontaneous preterm birth. STUDY DESIGN: Cervical and vaginal specimens for fetal fibronectin were obtained at 24, 26, 28, and 30 weeks' gestation from 2926 women. Quantitative fetal fibronectin values were calculated by using absorbances determined by enzyme-linked immunosorbent assay. The highest fetal fibronectin value (cervical or vaginal) for each woman at each visit was evaluated in relation to spontaneous preterm birth at <35 weeks' gestation. Receiver operating characteristic curves were constructed to determine the optimal cutoff point for fetal fibronectin values to predict spontaneous preterm birth at <35 weeks' gestation and within 4 weeks of testing. RESULTS: The risk of spontaneous preterm birth increased as a function of increasing fetal fibronectin values from approximately 20 to 300 ng/mL. Fetal fibronectin values greater than or equal to 300 ng/mL were not associated with a fur ther increase in spontaneous preterm birth. Examination of the receiver operating characteristic curve indicates that the optimal cutoff point for a positive fetal fibronectin test result at 24 to 30 weeks' gestation to predict spontaneous preterm birth at <35 weeks is between 45 and 60 ng/mL. CONCLUSION: Increasing levels of cervicovaginal fetal fibronectin up to 300 ng/mL are associated with an increasing risk of spontaneous preterm birth. Nevertheless, at 24 to 30 weeks, the value currently used, 50 ng of fetal fibronectin per milliliter appears to be a reasonable cutoff point for predicting spontaneous: preterm birth at <35 weeks' gestation. C1 Univ Alabama, Dept Obstet & Gynecol, Birmingham, AL 35249 USA. Univ Tennessee, Memphis, TN USA. Ohio State Univ, Columbus, OH 43210 USA. Wake Forest Univ, Winston Salem, NC 27109 USA. Univ Chicago, Chicago, IL 60637 USA. George Washington Univ, Washington, DC USA. Med Univ S Carolina, Charleston, SC 29425 USA. Univ Pittsburgh, Magee Womens Hosp, Pittsburgh, PA 15213 USA. Univ Oklahoma, Oklahoma City, OK USA. Univ Cincinnati, Cincinnati, OH USA. Wayne State Univ, Detroit, MI USA. NICHHD, Bethesda, MD 20892 USA. RP Goepfert, AR (reprint author), Univ Alabama, Dept Obstet & Gynecol, 619 S 19th St,Old Hillman Bldg,Room 450, Birmingham, AL 35249 USA. OI caritis, steve/0000-0002-2169-0712 FU NICHD NIH HHS [HD21410, HD21414, HD21434] NR 7 TC 43 Z9 43 U1 1 U2 6 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0002-9378 J9 AM J OBSTET GYNECOL JI Am. J. Obstet. Gynecol. PD DEC PY 2000 VL 183 IS 6 BP 1480 EP 1483 DI 10.1067/mob.2000.107067 PG 4 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 384NP UT WOS:000165952000036 PM 11120514 ER PT J AU Sibai, BM Caritis, SN Hauth, JC MacPherson, C VanDorsten, JP Klebanoff, M Landon, M Paul, RH Meis, PJ Miodovnik, M Dombrowski, MP Thurnau, GR Moawad, AH Roberts, J AF Sibai, BM Caritis, SN Hauth, JC MacPherson, C VanDorsten, JP Klebanoff, M Landon, M Paul, RH Meis, PJ Miodovnik, M Dombrowski, MP Thurnau, GR Moawad, AH Roberts, J CA Natl Inst Child Hlth Human Dev Mat TI Preterm delivery in women with pregestational diabetes mellitus or chronic hypertension relative to women with uncomplicated pregnancies SO AMERICAN JOURNAL OF OBSTETRICS AND GYNECOLOGY LA English DT Article DE hypertension; indicated preterm delivery; pregestational diabetes mellitus; spontaneous preterm delivery ID RISK-FACTORS; PREECLAMPSIA; PREDICTION; MORBIDITY; BIRTHS AB OBJECTIVE: The purpose of this study was to compare the rates of indicated and spontaneous preterm delivery among women with chronic hypertension or pregestational diabetes mellitus with the rates among healthy women. STUDY DESIGN: This was a secondary analysis of data from healthy women with singleton gestations enrolled in a prospective observational study for prediction of preterm delivery (control group, n = 2738), women-with pregestational diabetes mellitus requiring insulin therapy (n = 461), and women with chronic hypertension (n = 761). The two latter groups were enrolled in a randomized multicenter trial for prevention of preeclampsia. The main outcome measures were rates of preterm delivery, either spontaneous (preterm labor or rupture of membranes) or indicated (for maternal or fetal reasons), and neonatal outcomes. RESULTS: The overall rates of preterm delivery were significantly higher among women with diabetes mellitus (38%) and hypertension (33.1%) than among control women (13.9%). Rates were also significantly higher for delivery at <35 weeks' gestation. Women with diabetes mellitus had significantly higher rates of both indicated preterm delivery (21.9% vs 3.4%; odds ratio, 8.1;95% confidence interval, 6.0-10.9) and spontaneous preterm delivery (16.1% vs 10.5%; odds ratio, 1.6, 95% confidence interval, 1.2-2.2) than did women in the control group. In addition, they had significantly higher rates of both indicated preterm delivery (odds ratio, 4.8; 95% confidence interval, 3.0-7.5) and spontaneous preterm delivery (odds ratio, 2.1;95% confidence interval, 1.4-3.0) at <35 weeks' gestation than did control women. Compared with control women those with chronic hypertension had higher rates of indicated preterm delivery at both <37 weeks' gestation (21.9% vs 3.4%; odds ratio, 8.1; 95% confidence interval, 6.2-10.6) and at <35 weeks' gestation (12.1% vs 1.6%; odds ratio, 8.2; 95% confidence interval, 5.7-11.9), but there were no differences in rates of spontaneous preterm delivery. CONCLUSION: The increased Fate of preterm delivery among women with chronic hypertension relative to control women was primarily an increase in indicated preterm delivery, whereas the rates of both spontaneous and indicated preterm delivery were increased among women with pregestational diabetes mellitus. C1 Univ Tennessee, Dept Obstet & Gynecol, Memphis, TN USA. Univ Pittsburgh, Dept Obstet & Gynecol, Pittsburgh, PA USA. Univ Alabama, Dept Obstet & Gynecol, Birmingham, AL 35294 USA. George Washington Univ, Ctr Biostat, Dept Obstet & Gynecol, Washington, DC USA. Med Univ S Carolina, Dept Obstet & Gynecol, Charleston, SC 29425 USA. NICHHD, Dept Obstet & Gynecol, Bethesda, MD 20892 USA. Univ Chicago, Dept Obstet & Gynecol, Chicago, IL 60637 USA. Ohio State Univ, Dept Obstet & Gynecol, Columbus, OH 43210 USA. Univ Cincinnati, Coll Med, Dept Obstet & Gynecol, Cincinnati, OH 45267 USA. Univ So Calif, Dept Obstet & Gynecol, Los Angeles, CA 90089 USA. Wake Forest Univ, Dept Obstet & Gynecol, Winston Salem, NC 27109 USA. Wayne State Univ, Dept Obstet & Gynecol, Detroit, MI USA. Univ Oklahoma, Norman, OK 73019 USA. RP Sibai, BM (reprint author), Univ Cincinnati, Coll Med, Dept Obstet & Gynecol, 231 Bethesda Ave,Rm 4415, Cincinnati, OH 45267 USA. OI caritis, steve/0000-0002-2169-0712 FU NICHD NIH HHS [HD 19897, HD21410, HD21414] NR 13 TC 74 Z9 75 U1 1 U2 4 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0002-9378 J9 AM J OBSTET GYNECOL JI Am. J. Obstet. Gynecol. PD DEC PY 2000 VL 183 IS 6 BP 1520 EP 1524 DI 10.1067/mob.2000.107621 PG 5 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 384NP UT WOS:000165952000043 PM 11120521 ER PT J AU Landuzzi, L De Giovanni, C Nicoletti, G Rossi, I Ricci, C Astolfi, A Scopece, L Scotlandi, K Serra, M Bagnara, GP Nanni, P Lollini, PL AF Landuzzi, L De Giovanni, C Nicoletti, G Rossi, I Ricci, C Astolfi, A Scopece, L Scotlandi, K Serra, M Bagnara, GP Nanni, P Lollini, PL TI The metastatic ability of Ewing's sarcoma cells is modulated by stem cell factor and by its receptor c-kit SO AMERICAN JOURNAL OF PATHOLOGY LA English DT Article ID GROWTH-FACTOR; NEUROECTODERMAL TUMOR; HUMAN OSTEOSARCOMA; AUTOCRINE GROWTH; TYROSINE KINASE; MAST-CELLS; LIGAND; EXPRESSION; LINES; FIBROBLASTS AB Ewing's sarcoma is a primitive highly malignant tumor of bone and soft tissues usually metastasizing to bone, bone marrow, and lung. Growth factor receptors and their ligands may be involved in its growth and dissemination. We analyzed the expression of c-kit and its ligand stem cell factor (SCF) in a panel of six Ewing's sarcoma cell lines. All cell lines exhibited substantial levels of surface c-kit expression, and five of six displayed transmembrane SCP on the cell surface. Expression of c-kit was down-modulated in all lines by exposure to exogenous SCF. The SCF treatment was able to confer to cells a growth advantage in vitro, due both to an increase in cell proliferation and to a reduction in the apoptotic rate. When used in the lower compartment of a migration chamber, SCF acted as a strong chemoattractant for Ewing's sarcoma cells. The pretreatment of cells with SCF reduced their chemotactic response to SCF. in athymic nude mice, Ewing's sarcoma cells injected intravenously metastasized to the lung and to a variety of extrapulmonary sites, including bone and bone marrow. Metastatic sites resembled those observed in Ewing's sarcoma patients and corresponded to SCF-rich microenvironments. The in vitro pretreatment of cells with SCF strongly reduced the metastatic ability of Ewing's sarcoma cells, both to the lung and to extrapulmonary sites. This could be dependent on the down-modulation of c-kit expression observed in SCF-pretreated cells, leading to a reduced sensitivity to the chemotactic and proliferative actions of SCF. Our results indicate that the response to SCF mediated by c-kit may be involved in growth, migration, and metastatic ability of Ewing's sarcoma cells. C1 Univ Bologna, Dept Expt Pathol, I-40126 Bologna, Italy. Univ Bologna, Sect Canc Res, Bologna, Italy. Univ Bologna, Inst Histol & Gen Embryol, Bologna, Italy. Univ Bologna, Interdept Canc Res Ctr G Prodi, Bologna, Italy. Natl Canc Inst, Genoa, Italy. Rizzoli Inst, Biotechnol Satellite Unit, Bologna, Italy. Rizzoli Inst, Lab Oncol Res, Bologna, Italy. RP Landuzzi, L (reprint author), Univ Bologna, Dipartimento Patol Sperimentale, Sez Cancerol, Viale Fiiopanti 22, I-40126 Bologna, Italy. RI Lollini, Pier Luigi/A-7644-2008; De Giovanni, Carla/B-1312-2009; Serra, Massimo/J-4878-2016; OI Lollini, Pier Luigi/0000-0003-1702-4108; Serra, Massimo/0000-0003-0742-1177; Nanni, Patrizia/0000-0001-5319-0803; astolfi, annalisa/0000-0002-2732-0747 NR 44 TC 52 Z9 55 U1 0 U2 0 PU AMER SOC INVESTIGATIVE PATHOLOGY, INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3993 USA SN 0002-9440 J9 AM J PATHOL JI Am. J. Pathol. PD DEC PY 2000 VL 157 IS 6 BP 2123 EP 2131 DI 10.1016/S0002-9440(10)64850-X PG 9 WC Pathology SC Pathology GA 379XK UT WOS:000165668300039 PM 11106584 ER PT J AU Gladwin, MT Yao, XL Cowan, M Huang, XL Schneider, R Grant, LR Logun, C Shelhamer, JH AF Gladwin, MT Yao, XL Cowan, M Huang, XL Schneider, R Grant, LR Logun, C Shelhamer, JH TI Retinoic acid reduces p11 protein levels in bronchial epithelial cells by a posttranslational mechanism SO AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY LA English DT Article DE annexin II light chain; cytosolic phospholipase A(2); arachidonic acid ID ACUTE PROMYELOCYTIC LEUKEMIA; PHOSPHOLIPASE A(2); INHIBITION; DEXAMETHASONE; PROTEASOME; SYNTHASE AB p11 is a member of the S100 family of proteins, is the cellular ligand of annexin II, and interacts with the carboxyl region of 85-kDa cytosolic phospholipase A(2) (cPLA(2)), inhibiting cPLA(2) activity and arachidonic acid (AA) release. We studied the effect of retinoic acid (RA) on PLA(2) activity in human bronchial epithelial cells and whether p11 contributes to these effects. The addition of 10(-6) M RA resulted in reduced p11 protein levels at 4 days, with the greatest effect observed on days 6 and 7. This effect was dose related (10(-6) to 10(-9) M). RA treatment (10(-6) M) had no effect on cPLA(2) protein levels. p11 mRNA levels were unchanged at 6 and 8 days of treatment (correlating with maximum p11 protein reduction). Treatment with RA reduced p11 levels in control cells and in cells transfected with a p11 expression vector, suggesting a posttranslational mechanism. Lactacystin (10(-6) M), an inhibitor of the human 26S proteasome, blocked the decrease in p11 observed with RA treatment. Compared with control cells (n = 3), RA-treated cells (n = 3) had significantly increased AA release after treatment with the calcium ionophore A-23187 (P = 0.006). Therefore, RA reduces p11 protein expression and increases PLA(2) activity and AA release. C1 NIH, Warren G Magnuson Clin Ctr, Dept Crit Care Med, Bethesda, MD 20892 USA. RP Gladwin, MT (reprint author), NIH, Warren G Magnuson Clin Ctr, Dept Crit Care Med, Bldg 10,Rm 7D43,10 Ctr Dr,MSC 1662, Bethesda, MD 20892 USA. NR 17 TC 11 Z9 11 U1 0 U2 4 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 1040-0605 J9 AM J PHYSIOL-LUNG C JI Am. J. Physiol.-Lung Cell. Mol. Physiol. PD DEC PY 2000 VL 279 IS 6 BP L1103 EP L1109 PG 7 WC Physiology; Respiratory System SC Physiology; Respiratory System GA 375GY UT WOS:000165392200011 PM 11076800 ER PT J AU Promeneur, D Kwon, TH Yasui, M Kim, GH Frokiaer, J Knepper, MA Agre, P Nielsen, S AF Promeneur, D Kwon, TH Yasui, M Kim, GH Frokiaer, J Knepper, MA Agre, P Nielsen, S TI Regulation of AQP6 mRNA and protein expression in rats in response to altered acid-base or water balance SO AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY LA English DT Article DE aquaporin-6; intercalated cell; principal cell ID CORTICAL COLLECTING DUCT; INTERCALATED CELLS; METABOLIC-ACIDOSIS; SODIUM-CHLORIDE; KIDNEY; CHANNEL; AQUAPORINS; PERMEABILITY; PHYSIOLOGY; MEMBRANE AB In the rat, aquaporin-6 (AQP6) is mainly localized in intercalated cells (ICs) in collecting ducts, where it is exclusively associated with intracellular vesicles. In this study, we examined whether AQP6 protein and mRNA expression were regulated in the inner medulla or inner stripe of the outer medulla. Rats treated with dietary alkali or acid load for 7 days with a fixed daily water intake revealed appropriate changes in urine pH but unchanged urine output. AQP6 protein and mRNA abundance were increased in alkali-loaded rats (187 +/- 18 and 151 +/- 17% of control, respectively), whereas no changes were observed in acid-loaded rats. Immunohistochemistry revealed increased IC AQP6 labeling in alkali-loaded rats but not in acid-loaded rats. In contrast, administration of NH(4)Cl in the drinking water for 2 wk (free access to water) revealed a significant increase in AQP6 protein abundance (194 +/- 9% of control), but this was associated with increased water intake. Combined, this suggests that AQP6 expression was not affected by acid loading per se but rather was in response to changes in water intake. Consistent with this, water loading for 48 h was associated with increased AQP6 protein abundance, compared with thirsted rats. Moreover, rats with lithium-induced nephrogenic diabetes insipidus had a threefold increase in both AQP6 protein and mRNA expression. Overall, these results suggest that AQP6 expression in collecting duct ICs is regulated by altered acid/alkali load or water balance. Thus AQP6 may contribute to maintenance of acid-base homeostasis and water balance. C1 Univ Aarhus, Dept Cell Biol, Inst Anat, DK-8000 Aarhus C, Denmark. Johns Hopkins Univ, Sch Med, Dept Med & Biol Chem, Baltimore, MD 21205 USA. NHLBI, Kidney & Electrolyte Metab Lab, NIH, Bethesda, MD 20892 USA. Aarhus Univ Hosp, Dept Clin Physiol, DK-8200 Aarhus N, Denmark. Aarhus Univ Hosp, Inst Expt Clin Res, DK-8200 Aarhus N, Denmark. RP Kwon, TH (reprint author), Univ Aarhus, Dept Cell Biol, Inst Anat, DK-8000 Aarhus C, Denmark. EM sn@ana.au.dk FU Intramural NIH HHS [Z01 HL001285-21, Z99 HL999999] NR 49 TC 19 Z9 19 U1 0 U2 1 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 1931-857X J9 AM J PHYSIOL-RENAL JI Am. J. Physiol.-Renal Physiol. PD DEC PY 2000 VL 279 IS 6 BP F1014 EP F1026 PG 13 WC Physiology; Urology & Nephrology SC Physiology; Urology & Nephrology GA 377UR UT WOS:000165545100005 PM 11097619 ER PT J AU Helfand, WH Lazarus, J Theerman, P AF Helfand, WH Lazarus, J Theerman, P TI "Children's clinic," by Mabel Dwight SO AMERICAN JOURNAL OF PUBLIC HEALTH LA English DT Article C1 Natl Lib Med, Hist Med Div, Bethesda, MD 20894 USA. RP Theerman, P (reprint author), Natl Lib Med, Hist Med Div, 8600 Rockville Pike, Bethesda, MD 20894 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER PUBLIC HEALTH ASSOC INC PI WASHINGTON PA 1015 FIFTEENTH ST NW, WASHINGTON, DC 20005 USA SN 0090-0036 J9 AM J PUBLIC HEALTH JI Am. J. Public Health PD DEC PY 2000 VL 90 IS 12 BP 1834 EP 1834 DI 10.2105/AJPH.90.12.1834 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 377KH UT WOS:000165512400004 PM 11144304 ER PT J AU Doody, MM Chimes, K AF Doody, MM Chimes, K TI The Social Security Administration "presumed living" search SO AMERICAN JOURNAL OF PUBLIC HEALTH LA English DT Letter C1 NCI, Div Canc Epidemiol & Genet, Radiat Epidemiol Branch, Bethesda, MD 20892 USA. Westat Inc, Rockville, MD USA. RP Doody, MM (reprint author), NCI, Div Canc Epidemiol & Genet, Radiat Epidemiol Branch, 6120 Execut Blvd,Execut Plaza S,Room 7088, Bethesda, MD 20892 USA. FU NCI NIH HHS [N02-CP-81121] NR 4 TC 2 Z9 2 U1 0 U2 0 PU AMER PUBLIC HEALTH ASSOC INC PI WASHINGTON PA 1015 FIFTEENTH ST NW, WASHINGTON, DC 20005 USA SN 0090-0036 J9 AM J PUBLIC HEALTH JI Am. J. Public Health PD DEC PY 2000 VL 90 IS 12 BP 1948 EP 1949 DI 10.2105/AJPH.90.12.1948 PG 2 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 377KH UT WOS:000165512400031 PM 11111276 ER PT J AU Miller, DL AF Miller, DL TI Angiography in polyarteritis nodosa SO AMERICAN JOURNAL OF ROENTGENOLOGY LA English DT Letter C1 Uniformed Serv Univ Hlth Sci, F Edward Herbert Sch Med, Bethesda, MD 20814 USA. USN, Med Ctr, Bethesda, MD 20889 USA. NCI, Bethesda, MD 20889 USA. RP Miller, DL (reprint author), USN, Med Ctr, Bethesda, MD 20889 USA. NR 4 TC 11 Z9 11 U1 0 U2 1 PU AMER ROENTGEN RAY SOC PI RESTON PA 1891 PRESTON WHITE DR, SUBSCRIPTION FULFILLMENT, RESTON, VA 22091 USA SN 0361-803X J9 AM J ROENTGENOL JI Am. J. Roentgenol. PD DEC PY 2000 VL 175 IS 6 BP 1747 EP 1747 PG 1 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 376JN UT WOS:000165454600051 PM 11090415 ER PT J AU Middleton, LP Palacios, DM Bryant, BR Krebs, P Otis, CN Merino, MJ AF Middleton, LP Palacios, DM Bryant, BR Krebs, P Otis, CN Merino, MJ TI Pleomorphic lobular carcinoma: Morphology, immunohistochemistry, and molecular analysis SO AMERICAN JOURNAL OF SURGICAL PATHOLOGY LA English DT Article DE breast; pleomorphic lobular carcinoma; prognostic markers; Her 2; p53 ID SIGNET-RING CELLS; ONCOGENE PROTEIN; BREAST-CARCINOMA; DNA ANALYSIS; VARIANTS; FEATURES; THERAPY; CANCER AB Infiltrating pleomorphic lobular carcinoma (PLC) is an aggressive variant of infiltrating lobular carcinoma. Recently, in situ changes identical to PLC (PLCIS) have been described. The role of prognostic markers and their correlation with therapeutics, clinical outcome, and genetic changes is not well established in PLC. The authors examined 38 cases of this entity to understand better this tumor's biology. Immunohistochemical (IHC) analysis was performed in 21 specimens for estrogen and progesterone steroid receptors, p53, Her 2 (p185), and GCDFP-15. Genomic deoxyribonucleic acid was obtained from microdissected tumor as well as normal control cells, and loss of heterozygosity was investigated at the ESR (16q24), p53 (TP53 17p), Her 2 (17q 11-12), and BRCA 1 (17q12-25) loci. In this series, the average patient age was 57.5 years (age range, 24-92 years). Twenty-seven women were postmenopausal. Tumor size ranged from 1.2 to 25 cm. Six patients were a pathologic stage I; 19, stage II; 12, stage III; and one, stage IV. Histologically, multifocal nodular aggregates of discohesive pleomorphic tumor cells were seen interspersed in dense and fibrotic breast parenchyma. Twenty-nine percent of the specimens demonstrated associated signet ring cells. The remainder had dishesive, globoid, plasmacytoid cells with high-grade nuclear features. PLCIS was identified in 17 of 38 patients (45%), and lobular carcinoma in situ (LCIS) was noted in 8 patients (21%). IHC analysis showed estrogen immunoreactivity in 81%, progesterone in 67%, GCDFP-15 in 71%, and Her 2 in 81% (2+ to 3+ membranous staining) of specimens. Antibodies to p53 stained the tumor cell nuclei in 48% of the tumors. Loss of heterozygosity was identified in 52% of the specimens at the p53 locus, 18% at the ESR locus, 19% to 24% at the Her 2 loci, and 27% to 32% at the BRCA 1 locus. Follow-up was available in 19 patients and ranged from 12 months to 15 years (mean, 73 months). Seven patients had no evidence of disease at last examination (range, 1-15 years), three patients were alive with disease (range, 2-14 years), and nine patients were dead of disease (range, 2 months-9 years). Six patients had subsequent diagnoses of tumor in the contralateral breast. Analysis shows that PLC tends to appear in older postmenopausal women who present with locally advanced disease. PLCIS was found to be associated with PLC 45% of the time, The aggressive clinical course of patients with PLC is supported by tumor immunoreactivity with unfavorable markers Her 2 and p53. Overexpression of Her 2 in PLC may be therapeutically relevant, enabling the use of novel chemotherapeutic drugs like Herceptin. Interestingly, tumors that were Her 2, immunoreactive also maintained estrogen hormone immunoreactivity. C1 NCI, Bethesda, MD 20892 USA. Tufts Univ, Sch Med, Baystate Med Ctr, Springfield, MA 01199 USA. RP Middleton, LP (reprint author), Univ Texas, MD Anderson Canc Ctr, Dept Pathol, Box 85,1515 Holcombe Blvd, Houston, TX 77030 USA. NR 34 TC 95 Z9 96 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0147-5185 J9 AM J SURG PATHOL JI Am. J. Surg. Pathol. PD DEC PY 2000 VL 24 IS 12 BP 1650 EP 1656 DI 10.1097/00000478-200012000-00009 PG 7 WC Pathology; Surgery SC Pathology; Surgery GA 378NL UT WOS:000165590400009 PM 11117786 ER PT J AU Maglia, A Buchholz, DR AF Maglia, A Buchholz, DR TI Heterochrony and patterns of osteogenesis in pelobatoid frogs. SO AMERICAN ZOOLOGIST LA English DT Meeting Abstract C1 Univ Kansas, Lawrence, KS 66045 USA. NICHD, LME, NIH, Bethesda, MD USA. NR 0 TC 0 Z9 0 U1 0 U2 2 PU AMER SOC ZOOLOGISTS PI LAWRENCE PA 1041 NEW HAMPSHIRE ST, LAWRENCE, KS 66044 USA SN 0003-1569 J9 AM ZOOL JI Am. Zool. PD DEC PY 2000 VL 40 IS 6 BP 1113 EP 1114 PG 2 WC Zoology SC Zoology GA 422RA UT WOS:000168132000491 ER PT J AU Shi, YB AF Shi, YB TI Mechanism and developmental function of gene regulation by thyroid hormone receptors in Xenopus laevis. SO AMERICAN ZOOLOGIST LA English DT Meeting Abstract C1 NICHD, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER SOC ZOOLOGISTS PI LAWRENCE PA 1041 NEW HAMPSHIRE ST, LAWRENCE, KS 66044 USA SN 0003-1569 J9 AM ZOOL JI Am. Zool. PD DEC PY 2000 VL 40 IS 6 BP 1211 EP 1211 PG 1 WC Zoology SC Zoology GA 422RA UT WOS:000168132000735 ER PT J AU Wellems, TE Evans, AG AF Wellems, TE Evans, AG TI Coevolutionary genetics of Plasmodium malaria parasites and their human hosts SO AMERICAN ZOOLOGIST LA English DT Meeting Abstract C1 NIAID, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER SOC ZOOLOGISTS PI LAWRENCE PA 1041 NEW HAMPSHIRE ST, LAWRENCE, KS 66044 USA SN 0003-1569 J9 AM ZOOL JI Am. Zool. PD DEC PY 2000 VL 40 IS 6 BP 1254 EP 1255 PG 2 WC Zoology SC Zoology GA 422RA UT WOS:000168132000847 ER PT J AU Raychaudhuri, S Karmakar, P Thakur, AR AF Raychaudhuri, S Karmakar, P Thakur, AR TI gamma-Ray-induced DNA damage and repair in Methanosarcina barkeri SO ANAEROBE LA English DT Article DE Methanosarcina barkeri; bromoethanesulfonate; 3-aminobenzamide; arabinosine-CTP; repair ID DOUBLE-STRAND BREAKS; MAMMALIAN-CELLS; POLY(ADP-RIBOSE) POLYMERASE; HALOBACTERIUM-CUTIRUBRUM; ARCHAEA; METHANOGENESIS; BACTERIA; PROTEINS; ACETATE; GROWTH AB The capacity of the mesophilic archaeon, Methanosarcina barkeri (DSM 804) for DNA double strand break repair following Co-60-gamma irradiation was investigated. The genome (1.9 Mb) of Methanosarcina barkeri was largely fragmented and was found to be repaired on incubation in medium under anaerobic conditions at 37 degreesC for 4 h. To get an insight into its repair process a set of inhibitors were used. The methanogenesis inhibitor, bromoethane-sulfonate showed partial inhibition of repair in Methanosarcina barkeri but not in Escherichia coli or human peripheral blood mononuclear cells. The Methanosarcina barkeri cells could also partially repair the DNA damage in a non-nutrient medium. Arabinosine-CTP, a nucleoside analogue and a polymerase inhibitor, completely inhibited repair in this archaeon. Arabinosine-CTP did not affect DSB (double-strand break) repair in human peripheral blood mononuclear cells but completely inhibited repair in Escherichia coli (a bacterium). The involvement of polymerase indicates recombination to be the underlying mechanism in DSB repair of Methanosarcina barkeri. 3-Aminobenzamide, a poly (ADP-ribose) polymerase inhibitor, completely inhibited repair in this archaeon as well as in eukarya but not in Escherichia coli showing the involvement of poly (ADP-ribose) polymerase in the DSB repair of Methanosarcina barkeri. (C) 2000 Academic Press. C1 Univ Coll Sci & Technol, Dept Biophys Mol Biol & Genet, Calcutta 700009, W Bengal, India. NIA, GRC, NIH, Baltimore, MD 21224 USA. RP Thakur, AR (reprint author), Univ Coll Sci & Technol, Dept Biophys Mol Biol & Genet, 92 APC Rd, Calcutta 700009, W Bengal, India. NR 42 TC 3 Z9 3 U1 0 U2 1 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 1075-9964 J9 ANAEROBE JI Anaerobe PD DEC PY 2000 VL 6 IS 6 BP 325 EP 331 DI 10.1006/anae.2000.0359 PG 7 WC Microbiology SC Microbiology GA 391HF UT WOS:000166350000002 ER PT J AU Roth, SM Martel, GF Ivey, FM Lemmer, JT Metter, EJ Hurley, BF Rogers, MA AF Roth, SM Martel, GF Ivey, FM Lemmer, JT Metter, EJ Hurley, BF Rogers, MA TI Skeletal muscle satellite cell populations in healthy young and older men and women SO ANATOMICAL RECORD LA English DT Article DE aging; electron microscopy; mitochondria; morphology; myonuclei; ultrastructure ID SLOW-TONIC MUSCLE; BODY-COMPOSITION; RAT MUSCLE; AGE; HYPERTROPHY; REGENERATION; STRENGTH; RECOVERY; ULTRASTRUCTURE; PROLIFERATION AB The purpose of the present investigation was to assess satellite cell populations and morphology in m. vastus lateralis biopsies obtained from young (20-30 years) and older (65-75 years) healthy, sedentary men and women. Multiple muscle biopsies were obtained from 14 young individuals (men, n = 7; women, n = 7) and 15 older individuals (men, n = 8; women, n = 7). Muscle fibers were viewed longitudinally using a Zeiss EM 10 CA electron microscope. All myonuclei and satellite cells were counted and satellite cells were photographed for morphological analysis. The proportion of satellite cells [satellite cells/(myonuclei + satellite cells)] did not differ among the four subject groups (1.7-2.8%), nor did proportions differ when subject groups were combined for age and gender comparisons. Few morphological differences were noted between groups; however, lipofuscin granules were more prominent in satellite cells from older subjects and women demonstrated significantly larger satellite cell and satellite cell nucleus areas than men. Mitochondria from satellite cells (regardless of group) were more pallid and exhibited fewer cristae than mitochondria located in the adjacent muscle fiber. The results of the current investigation suggest that, despite findings in animal models, satellite cell populations are not significantly lower in healthy, sedentary older compared to young adult men and women. Anat Rec 260:351-358, 2000. (C) 2000 Wiley-Liss, Inc. C1 Univ Maryland, Coll Hlth & Human Performance, Dept Kinesiol, College Pk, MD 20742 USA. Univ Maryland Eastern Shore, Dept Phys Therapy, Princess Anne, MD 21853 USA. NIA, Gerontol Res Ctr, Baltimore, MD 21224 USA. RP Roth, SM (reprint author), Univ Pittsburgh, Dept Human Genet, A218 Crabtree Hall, Pittsburgh, PA 15261 USA. OI Roth, Stephen/0000-0002-7841-3695 FU NIA NIH HHS [AG-00268, AG-42148] NR 43 TC 83 Z9 87 U1 0 U2 4 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0003-276X J9 ANAT REC JI Anat. Rec. PD DEC 1 PY 2000 VL 260 IS 4 BP 351 EP 358 DI 10.1002/1097-0185(200012)260:4<350::AID-AR30>3.0.CO;2-6 PG 8 WC Anatomy & Morphology SC Anatomy & Morphology GA 379XR UT WOS:000165668900003 PM 11074399 ER PT J AU Helmers, KF Baker, B O'Kelly, B Tobe, S AF Helmers, KF Baker, B O'Kelly, B Tobe, S TI Anger expression, gender, and ambulatory blood pressure in mild, unmedicated adults with hypertension SO ANNALS OF BEHAVIORAL MEDICINE LA English DT Article ID WHITE-COAT HYPERTENSION; STRESS; DISCRIMINATION; PERSONALITY; PREVALENCE; PREDICTORS; DETROIT AB The suppression of anger has been associated with the development of hypertension. This study evaluated the association between anger management style (anger-in and anger-out) and ambulatory blood pressure (ABP) in patients with repented clinic diastolic blood pressures (DBPs) between 90-105 mmHg, unmedicated and with no known coronary artery disease. A total of 128 men (46.0 years) and 66 women (46.6 years) participated. Fourteen percent of men and 35% of women were classified as having "white coat" hypertension (daytime DBP <85 mmHg). Mean awake and sleep DBP and systolic blood pressure (SBP) were evaluated in a repeated measures analysis of variance (ANOVA). Anger-in and anger-oct scores were categorized into low, medium, and high t-scores (<50, 50-59, greater than or equal to 60). Results indicated that in women, increasing anger-in is associated with greater SBPs while awake and sleeping, whereas no effect was found for DBP, nor any effect in men. No significant association was found between gender anger-out, and ABP. The clinical diagnostic status of white coat hypertension was not differentially associated with anger-in or anger-out in men and women. In conclusion in a sample of mild unmedicated adults with hypertension, suppression of anger is associated with greater ambulatory SEP in women, but not in men. C1 York Univ, N York, ON M3J 1P3, Canada. Toronto Hosp, Toronto, ON M5T 2S8, Canada. Sunnybrook Hlth Sci Ctr, Toronto, ON, Canada. RP Helmers, KF (reprint author), NINR, NIH, Bldg 45,Room 3AN12,45 Ctr Dr, Bethesda, MD 20892 USA. NR 24 TC 17 Z9 17 U1 1 U2 3 PU SOC BEHAVIORAL MEDICINE PI MIDDLETON PA 7611 ELMWOOD AVE, STE 201, MIDDLETON, WI 53562-3161 USA SN 0883-6612 J9 ANN BEHAV MED JI Ann. Behav. Med. PD WIN PY 2000 VL 22 IS 1 BP 60 EP 64 DI 10.1007/BF02895168 PG 5 WC Psychology, Multidisciplinary SC Psychology GA 345EQ UT WOS:000088802800007 PM 10892529 ER PT J AU Major, EO AF Major, EO TI From telomeres to T-antigens: Many roads ... multiple pathways ... novel associations in the search for the origins of human gliomas SO ANNALS OF NEUROLOGY LA English DT Editorial Material ID JC-VIRUS; POLYOMAVIRUS; CELLS C1 NINDS, Lab Mol Med & Neurosci, Bethesda, MD 20892 USA. RP Major, EO (reprint author), NINDS, Lab Mol Med & Neurosci, Bldg 36,Rm 4D04, Bethesda, MD 20892 USA. NR 15 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0364-5134 J9 ANN NEUROL JI Ann. Neurol. PD DEC PY 2000 VL 48 IS 6 BP 823 EP 825 PG 3 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 378WD UT WOS:000165607500001 PM 11117537 ER PT J AU Lippa, CF Zhukareva, V Kawarai, T Uryu, K Shafiq, M Nee, LE Grafman, J Liang, Y St George-Hyslop, PH Trojanowski, JQ Lee, VMY AF Lippa, CF Zhukareva, V Kawarai, T Uryu, K Shafiq, M Nee, LE Grafman, J Liang, Y St George-Hyslop, PH Trojanowski, JQ Lee, VMY TI Frontotemporal dementia with novel tau pathology and a Glu342Val tau mutation SO ANNALS OF NEUROLOGY LA English DT Article ID ALZHEIMERS-DISEASE; PRESENILE-DEMENTIA; GENE; PHOSPHORYLATION; MISSENSE; FTDP-17; SITES; CHROMOSOME-17 AB It is unclear how tau gene mutations cause frontotemporal dementia (FTD) with parkinsonism linked to chromosome 17 (FTDP-17), but those in exon 10 (E10) or the following intron may be pathogenic by altering E10 splicing, perturbing the normal 1:1 ratio of four versus three microtubule-binding repeat tau (4R:3R tau ratio) and forming tau inclusions. We report on a 55-year old woman with frontotemporal dementia and a family history of FTDP-17 in whom we found a novel E12 (Glu342Val) tau gene mutation, prominent frontotemporal neuron loss, intracytoplasmic tau aggregates, paired helical tau filaments, increased 4R tau messenger RNA, increased 4R tau without E2 or E3 inserts, decreased 4R tau with these inserts, and a 4R:3R tan ratio greater than 1 in gray and white matter. Thus, this novel Glu342Val mutation may cause FTDP-17 by unprecedented mechanisms that alter splicing of E2, E3, and E10 to preferentially increase 4R tau without amino terminal inserts and promote aggregation of tau filaments into cytopathic inclusions. C1 Med Coll Penn & Hahnemann Univ, Dept Neurol, Philadelphia, PA 19129 USA. Univ Penn, Sch Med, Dept Pathol & Lab Med, Ctr Neurodegenerat Dis Res, Philadelphia, PA 19104 USA. Univ Toronto, Dept Med, Ctr Res Neurodegenerat Dis, Toronto, ON, Canada. Univ Hlth Network, Dept Med Neurol, Toronto, ON, Canada. NIH, Cognit Neurosci Ctr, Bethesda, MD 20892 USA. RP Lippa, CF (reprint author), Med Coll Penn & Hahnemann Univ, Dept Neurol, 3300 Henry Ave, Philadelphia, PA 19129 USA. OI Grafman, Jordan H./0000-0001-8645-4457 NR 20 TC 66 Z9 70 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0364-5134 J9 ANN NEUROL JI Ann. Neurol. PD DEC PY 2000 VL 48 IS 6 BP 850 EP 858 DI 10.1002/1531-8249(200012)48:6<850::AID-ANA5>3.3.CO;2-M PG 9 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 378WD UT WOS:000165607500005 PM 11117541 ER PT J AU Fischer, JS Priore, RL Jacobs, LD Cookfair, DL Rudick, RA Herndon, RM Richert, JR Salazar, AM Goodkin, DE Granger, CV Simon, JH Grafman, JH Lezak, MD Hovey, KMO Perkins, KK Barilla-Clark, D Schacter, M Shucard, DW Davidson, AL Wende, KE Bourdette, DN Kooijmans-Coutinho, MF AF Fischer, JS Priore, RL Jacobs, LD Cookfair, DL Rudick, RA Herndon, RM Richert, JR Salazar, AM Goodkin, DE Granger, CV Simon, JH Grafman, JH Lezak, MD Hovey, KMO Perkins, KK Barilla-Clark, D Schacter, M Shucard, DW Davidson, AL Wende, KE Bourdette, DN Kooijmans-Coutinho, MF CA Multiple Scerlosis Collaborative R TI Neuropsychological effects of interferon beta-1a in relapsing multiple sclerosis SO ANNALS OF NEUROLOGY LA English DT Article; Proceedings Paper CT 52nd Annual Meeting of the American-Acadmey-of-Neurology CY APR, 1998 CL MINNEAPOLIS, MINNESOTA SP Amer Acad Neurol ID PLACEBO-CONTROLLED TRIAL; MAGNETIC-RESONANCE; COGNITIVE IMPAIRMENT; DISEASE PROGRESSION; DOUBLE-BLIND; BRAIN ATROPHY; FOLLOW-UP; DISABILITY; PATTERNS; LESIONS AB Cognitive dysfunction is common in multiple sclerosis (MS), yet few studies have examined effects of treatment on neuropsychological (NP) performance. To evaluate the effects of interferon beta -1a (IFN beta -1a 30 mug administered intramuscularly once weekly [Avonex]) on cognitive function, a Comprehensive NP Battery was administered at baseline and week 104 to relapsing MS patients in the phase III study, 166 of whom completed both assessments. A Brief NP Battery was also administered at 6-month intervals. The primary NP outcome measure was 2-year change on the Comprehensive NP Battery, grouped into domains of information processing and learning/memory (set A), visuospatial abilities and problem solving (set B), and verbal abilities and attention span (set C). NP effects were most pronounced in cognitive domains vulnerable to MS: IFN beta -1a had a significant beneficial effect on the set A composite, with a favorable trend evident on set B. Secondary outcome analyses revealed significant between-group differences in slopes for Brief NP Battery performance and time to sustained deterioration in a Paced Auditory Serial Addition Test processing rate, favoring the IFN beta -1a group. These results support and extend previous observations of significant beneficial effects of IFN beta -1a for relapsing MS. C1 Cleveland Clin, Mellen Ctr Multiple Sclerosis Treatment & Res, Cleveland, OH 44106 USA. SUNY Buffalo, Buffalo, NY 14260 USA. Buffalo Gen Hosp, Buffalo, NY 14203 USA. Ctr Funct Assessment Res, Buffalo, NY USA. Univ Mississippi, Jackson, MS 39216 USA. Georgetown Med Ctr, Washington, DC USA. Walter Reed Army Med Ctr, Washington, DC 20307 USA. NINDS, Bethesda, MD 20892 USA. Univ Calif San Francisco, San Francisco, CA 94143 USA. Univ Colorado, Denver, CO 80202 USA. Oregon Hlth Sci Univ, Portland, OR 97201 USA. Biogen, Dept Med Res, Cambridge, MA USA. RP Fischer, JS (reprint author), 170 Fuller Lane, Winnetka, IL 60093 USA. OI Grafman, Jordan H./0000-0001-8645-4457 FU PHS HHS [R01-26321] NR 51 TC 204 Z9 206 U1 1 U2 4 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0364-5134 J9 ANN NEUROL JI Ann. Neurol. PD DEC PY 2000 VL 48 IS 6 BP 885 EP 892 DI 10.1002/1531-8249(200012)48:6<885::AID-ANA9>3.0.CO;2-1 PG 8 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 378WD UT WOS:000165607500009 PM 11117545 ER PT J AU Nguyen, DM Desai, S Chen, A Weiser, TS Schrump, DS AF Nguyen, DM Desai, S Chen, A Weiser, TS Schrump, DS TI Modulation of metastasis phenotypes of non-small cell lung cancer cells by 17-allylamino 17-demethoxy geldanamycin SO ANNALS OF THORACIC SURGERY LA English DT Article; Proceedings Paper CT 36th Annual Meeting of the Society-of-Thoracic-Surgeons CY JAN 31-FEB 02, 2000 CL FT LAUDERDALE, FLORIDA SP Soc Thorac Surg ID ENDOTHELIAL GROWTH-FACTOR; E-CADHERIN; PROGNOSTIC-SIGNIFICANCE; POOR-PROGNOSIS; BREAST-CANCER; TUMOR-CELLS; EXPRESSION; OVEREXPRESSION; MECHANISMS; ANTIBODIES AB Background. Cancer cells that overexpress c-erbB oncogenes exhibit resistance to chemotherapy, enhanced tumorigenicity, as well as increased propensity for metastasis. The aim of this study was to investigate if depletion of erbB-1/EGFR and erbB-2/HER2neu oncogene products by 17-allylamino 17-demethoxy Geldanamycin (17AAGA) could diminish the metastatic potential of non-small cell lung cancer (NSCLC) cells that express varying levels of the erbB1/erbB2 oncogenes. Methods. NSCLC cell lines (H460, H358, H322, or H661) were assayed for expression of erbB1 and erbB2, the cell adhesion molecule E-cadherin, secretion of the matrix metalloproteinase 9 (MMP-9), and vascular endothelial cell growth factor (VEGF), as well as their ability to invade Matrigel after 48-hour exposure to 17AAGA. Results. 17AAGA significantly depleted erbB1 or erbB2 levels in NSCLC cells expressing high levels of these proteins, and effectively inhibited their growth with IC,, values ranging from 50 to 90 nmol/L. Moreover, drug treatment enhanced E-cadherin expression in H322 and H358 cells, and inhibited secretion of MMP-9 and VEGF secretion by tumor cells. 17AAGA diminished hypoxia-induced upregulation of VEGF expression as well as growth factor-mediated augmentation of MMP-9 secretion, and profoundly inhibited the ability of H322 and H358 cells to migrate through Matrigel in response to chemoattractants. Conclusions. In addition to its known antiproliferative and chemosensitization effects, 17AAGA inhibits the metastatic phenotype of lung cancer cells. 17AAGA may be a novel pharmacologic agent for specific molecular intervention in lung cancer patients. (Ann Thorac Surg 2000;70:1853-60) ( C) 2000 by The Society of Thoracic Surgeons. C1 NCI, Div Clin Sci, Surg Branch, Thorac Oncol Sect,NIH, Bethesda, MD 20892 USA. RP Nguyen, DM (reprint author), NCI, Div Clin Sci, Surg Branch, Thorac Oncol Sect,NIH, Room 2B07,Bldg 10,10 Ctr Dr, Bethesda, MD 20892 USA. NR 25 TC 29 Z9 33 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0003-4975 J9 ANN THORAC SURG JI Ann. Thorac. Surg. PD DEC PY 2000 VL 70 IS 6 BP 1853 EP 1860 DI 10.1016/S0003-4975(00)01810-5 PG 8 WC Cardiac & Cardiovascular Systems; Respiratory System; Surgery SC Cardiovascular System & Cardiology; Respiratory System; Surgery GA 385UT UT WOS:000166022900018 PM 11156083 ER PT J AU Pettit, GR Minardi, MD Boyd, MR Pettit, RK AF Pettit, GR Minardi, MD Boyd, MR Pettit, RK TI Antineoplastic agents 463. Synthesis of combretastatin A-3 diphosphate prodrugs SO ANTI-CANCER DRUG DESIGN LA English DT Article DE cation salts; combretastatin A-3; diphosphate; prodrugs ID ASYMMETRIC-SYNTHESIS; DISODIUM PHOSPHATE; ANTITUMOR-ACTIVITY; TUMOR VASCULATURE; SOLID TUMORS; ANALOGS; POTENT; THERAPY; A4 AB A new and more efficient synthesis of combretastatin A-3 (2a) was completed (8.4% overall yield) starting from methyl gallate and isovanillin with aldehyde 5 and phosphonium salt 8 as key intermediates. Conversion of combretastatin A-3 (2a) to a series of diphosphate prodrugs (10a-1) was readily achieved. Both the diphosphate sodium (10a) and potassium salts (10c) displayed aqueous solubility in excess of 220 mg/ml at room temperature and good cancer cell line inhibitory activity. C1 Arizona State Univ, Canc Res Inst, Tempe, AZ 85287 USA. Arizona State Univ, Dept Chem & Biochem, Tempe, AZ 85287 USA. NCI, Lab Drug Discovery Res & Dev, DTP, DCTD,Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RP Pettit, RK (reprint author), Arizona State Univ, Canc Res Inst, POB 872404, Tempe, AZ 85287 USA. FU NCI NIH HHS [CA44344-05-12] NR 33 TC 12 Z9 12 U1 0 U2 3 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0266-9536 J9 ANTI-CANCER DRUG DES JI Anti-Cancer Drug Des. PD DEC PY 2000 VL 15 IS 6 BP 397 EP 403 PG 7 WC Biochemistry & Molecular Biology; Oncology; Chemistry, Medicinal; Pharmacology & Pharmacy SC Biochemistry & Molecular Biology; Oncology; Pharmacology & Pharmacy GA 467LV UT WOS:000170705400002 PM 11716432 ER PT J AU Qin, J Yang, YW Velyvis, A Gronenborn, A AF Qin, Jun Yang, Yanwu Velyvis, Algirdas Gronenborn, Angela TI Molecular Views of Redox Regulation: Three-Dimensional Structures of Redox Regulatory Proteins and Protein Complexes SO ANTIOXIDANTS & REDOX SIGNALING LA English DT Review AB The last decade has witnessed the explosion of research on redox-controlled cellular and biochemical Whereas the vast majority of these studies have centered on clinical, genetic, and biochemical aspects of redox signaling and regulation inside and outside the cell, a significant number of nuclear magnetic resonance (NMR) and crystallographic studies have been undertaken to obtain an atomic-level understanding of the mechanisms of the redox regulation. This review highlights the recent progress of three-dimensional structure determination of key proteins and protein complexes involved in redox regulation. An increased list of such class of protein structures and their complexes with ligands will provide invaluable insight into the molecular basis of redox-regulatory processes and may be useful for the future development of therapeutic agents for redox-related diseases. Antiox. Redox Signal. 2, 827-840. C1 [Qin, Jun; Yang, Yanwu; Velyvis, Algirdas] Cleveland Clin Fdn, Lerner Res Inst, Struct Biol Program, Cleveland, OH 44195 USA. [Gronenborn, Angela] NIDDK, Chem Phys Lab, NIH, Bethesda, MD USA. RP Qin, J (reprint author), Cleveland Clin Fdn, Lerner Res Inst NB20, Struct Biol Program, 9500 Euclid Ave, Cleveland, OH 44195 USA. EM qinj@ccf.org FU American Cancer Society; National Institutes of Health FX This work is supported in part by funding from the American Cancer Society and the National Institutes of Health to J.Q. NR 80 TC 8 Z9 9 U1 0 U2 1 PU MARY ANN LIEBERT, INC PI NEW ROCHELLE PA 140 HUGUENOT STREET, 3RD FL, NEW ROCHELLE, NY 10801 USA SN 1523-0864 EI 1557-7716 J9 ANTIOXID REDOX SIGN JI Antioxid. Redox Signal. PD WIN PY 2000 VL 2 IS 4 BP 827 EP U230 DI 10.1089/ars.2000.2.4-827 PG 15 WC Biochemistry & Molecular Biology; Endocrinology & Metabolism SC Biochemistry & Molecular Biology; Endocrinology & Metabolism GA V10ZC UT WOS:000207500900021 PM 11213487 ER PT J AU Nesterova, M Cho-Chung, YS AF Nesterova, M Cho-Chung, YS TI Oligonucleotide sequence-specific inhibition of gene expression, tumor growth inhibition, and modulation of cAMP signaling by an RNA-DNA hybrid antisense targeted to protein kinase A RI alpha subunit SO ANTISENSE & NUCLEIC ACID DRUG DEVELOPMENT LA English DT Article ID CYCLIC-AMP BINDING; REGULATORY SUBUNIT; MULTIDRUG-RESISTANCE; ANTITUMOR-ACTIVITY; OVARIAN-CANCER; HL-60 LEUKEMIA; BREAST-CANCER; MESSENGER-RNA; OLIGODEOXYNUCLEOTIDE; PHOSPHODIESTERASE AB The primary mediator of cAMP action in mammalian cells is cAMP-dependent protein kinase (PKA), There are two types of PKA, type I (PKA-I) and type II (PKA-II), which share a common catalytic subunit but contain distinct regulatory subunits, RI and RII, respectively. Evidence suggests that increased expression of RIalpha/PKA-I correlates with neoplastic cell growth. Here, we show that sequence-specific oligonucleotide inhibition of RIalpha expression results in inhibition of growth and modulation of cAMP signaling in cancer cells. The antisense promoted growth inhibition in a time-dependent, concentration-dependent, and sequence-dependent manner in human cancer cells in monolayer culture, and it inhibited colony formation in soft agar and tumor growth in nude mice. Among the cancer cells are LS-174T, HCT-15, and Cole-205 colon carcinoma cells; A-549 lung carcinoma cells; LNCaP prostate adenocarcinoma cells; Molt-4 leukemia cells; and Jurkat T lymphoma cells, Northern blot and immunoprecipitation analyses revealed that the growth inhibitory effect of the antisense correlated with a decrease in RIalpha expression at both the mRNA and protein levels. Pulse-chase experiments revealed that the antisense-directed inhibition of RIalpha expression resulted in compensatory changes in expression of the isoforms of R and C subunits and cAMP signaling in a cell type-specific manner, These results demonstrate that cAMP is ubiquitous in the regulation of cell growth and that the antisense oligonucleotide, which inhibits the synthesis of the RIalpha subunit of PKA, can be targeted to a single gene for treatment of cancer in a variety of cell types. C1 NCI, Cellular Biochem Sect, Tumor Immunol & Biol Lab, NIH, Bethesda, MD 20892 USA. RP Cho-Chung, YS (reprint author), NCI, Cellular Biochem Sect, Tumor Immunol & Biol Lab, NIH, Bldg 10,Room 5B05,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 52 TC 34 Z9 34 U1 0 U2 0 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1087-2906 J9 ANTISENSE NUCLEIC A JI Antisense Nucleic Acid Drug Dev. PD DEC PY 2000 VL 10 IS 6 BP 423 EP 433 DI 10.1089/oli.1.2000.10.423 PG 11 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Research & Experimental Medicine GA 389WZ UT WOS:000166262800003 PM 11198926 ER PT J AU Sedelnikova, OA Panyutin, IG Luu, AN Reed, MW Licht, T Gottesman, MM Neumann, RD AF Sedelnikova, OA Panyutin, IG Luu, AN Reed, MW Licht, T Gottesman, MM Neumann, RD TI Targeting the human mdr1 gene by I-125-labeled triplex-forming oligonucleotides SO ANTISENSE & NUCLEIC ACID DRUG DEVELOPMENT LA English DT Article ID KB CARCINOMA-CELLS; MULTIDRUG-RESISTANCE; MAMMALIAN-CELLS; P-GLYCOPROTEIN; DNA TRIPLEXES; EXPRESSION; SEQUENCE; DELIVERY; PEPTIDE; I-125 AB Antigene radiotherapy is our approach to targeting specific sites in the genome by combining the highly localized DNA damage produced by the decay of Auger electron emitters, such as I-125, With the sequence-specific action of tripler-forming oligonucleotides (TFO), As a model, we used the multidrug resistance gene (mdr1) overexpressed and amplified nearly 100 times in the human KB-V1 carcinoma cell line. Phosphodiester pyrrazolopyrinidine dG (BPG)-modified TFO complementary to the polypurine-polypyrimidine region of the mdr1 gene were synthesized and labeled with I-125-dCTP at the C5 position of two cytosines by the primer extension method,I-125-TFO were delivered into KB-V1 cells with several delivery systems. DNA from the I-125-TFO-treated cells was recovered and analyzed for sequence-specific cleavage in the mdr1 target by Southern hybridization, Experiments with plasmid DNA containing the mdr1 polypurine-polypyrimidine region and with purified genomic DNA confirmed the ability of the designed I-125-TFO to bind to and introduce double-strand breaks into the target sequence. We showed that I-125-TFO in nanomolar concentrations can recognize and cleave a target sequence in the mdr1 gene in situ, that is, within isolated nuclei and intact digitonin-permeabilized cells. Our results demonstrate the ability of I-125-TFO to target specific sequences in their natural environment, that is, within the eukaryotic nucleus, The nearly 100-fold amplification of the mdr1 gene in KB-V1 cells affords a very useful cell culture model for evaluation of methods to produce sequence-specific DNA double-strand breaks for gene-specific radiotherapy. C1 NCI, Dept Nucl Med, Warren G Magnuson Clin Ctr, NIH, Bethesda, MD 20892 USA. NCI, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. NCI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. Epoch Pharmaceut, Redmond, WA 98052 USA. RP Neumann, RD (reprint author), NCI, Dept Nucl Med, Warren G Magnuson Clin Ctr, NIH, Bldg 10,Room IC401,10 Ctr Dr MSC 1180, Bethesda, MD 20892 USA. NR 44 TC 24 Z9 26 U1 1 U2 4 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1087-2906 J9 ANTISENSE NUCLEIC A JI Antisense Nucleic Acid Drug Dev. PD DEC PY 2000 VL 10 IS 6 BP 443 EP 452 DI 10.1089/oli.1.2000.10.443 PG 10 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Research & Experimental Medicine GA 389WZ UT WOS:000166262800005 PM 11198928 ER PT J AU Bhargava, R Fernandez, DC Schaeberle, MD Levin, IW AF Bhargava, R Fernandez, DC Schaeberle, MD Levin, IW TI Effect of focal plane array cold shield aperture size on fourier transform infrared micro-imaging spectrometer performance SO APPLIED SPECTROSCOPY LA English DT Article DE fourier transform infrared (FT-IR); imaging; spectroscopy; focal plane array; polymer; signal-to-noise ratio; optimization ID DETECTOR; SPECTROSCOPY; TISSUES AB Micro-imaging spectrometers incorporating focal plane array (FPA) detection require careful demarcation of told shield aperture size for both optimal performance and prevention of errors. This study examines the effects of changing the diameter of the cold shield aperture on the intensity and spatial homogeneity of the incident radiation. A uniform polystyrene film was repeatedly imaged by using cold shields of varying aperture sizes. It is shown that a smaller than optimal aperture size leads to image edge clipping, resulting in an inefficient use of the array, lower overall signal, spectral distortions, and higher noise characteristics. Use of an aperture size larger than required causes a decrease in the effective dynamic range of measurements, resulting in higher noise levels. The advantages and necessity of optimizing imaging spectrometer performance by employing a cold shield with an appropriately sized aperture are discussed. C1 NIDDKD, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. RP Levin, IW (reprint author), NIDDKD, Chem Phys Lab, NIH, Bldg 2, Bethesda, MD 20892 USA. OI Bhargava, Rohit/0000-0001-7360-994X NR 17 TC 12 Z9 12 U1 0 U2 3 PU SOC APPLIED SPECTROSCOPY PI FREDERICK PA 201B BROADWAY ST, FREDERICK, MD 21701 USA SN 0003-7028 J9 APPL SPECTROSC JI Appl. Spectrosc. PD DEC PY 2000 VL 54 IS 12 BP 1743 EP 1750 DI 10.1366/0003702001949069 PG 8 WC Instruments & Instrumentation; Spectroscopy SC Instruments & Instrumentation; Spectroscopy GA 388NX UT WOS:000166188600004 ER PT J AU Jeong, KS Soh, Y Jeng, J Felder, MR Hardwick, JP Song, BJ AF Jeong, KS Soh, Y Jeng, J Felder, MR Hardwick, JP Song, BJ TI Cytochrome P450 2E1 (CYP2E1)-dependent production of a 37-kDa acetaldehyde-protein adduct in the rat liver SO ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS LA English DT Article DE acetaldehyde; acetaldehyde-protein adduct; alcohol metabolism; alcohol dehydrogenase; CYP2E1 inhibitor ID HYDROXYETHYL RADICAL ADDUCTS; LIPID-PEROXIDATION; ALDEHYDE DEHYDROGENASE; PLASMA-MEMBRANE; ETHANOL; ALCOHOL; CYP2E1; HEPATOCYTES; EXPRESSION; INDUCTION AB Ethanol-inducible cytochrome P450 2E1 (CYP2E1) has been shown to be involved in the metabolism of both ethanol and acetaldehyde, Acetaldehyde, produced from ethanol metabolism, is highly reactive and can form various protein adducts, In this study, we investigated the role of CYP2E1 in the production of a 37-kDa acetaldehyde-protein adduct, Rats were pair-fed an isocaloric control or an alcohol liquid diet with and without cotreatment of YH439, an inhibitor of CYP2E1 gene transcription, for 4 weeks. The soluble proteins from rat livers of each group were separated on SDS-polyacrylamide gels followed by immunoblot analysis using specific antibodies against the 37-kDa protein acetaldehyde adduct, In addition, catalytic activities of the enzymes involved in alcohol and acetaldehyde metabolism were measured and compared with the adduct level, Immunoblot analysis revealed that the 37-kDa adduct, absent in the pair-fed control, was evident in alcohol-fed rats but markedly reduced by YH439 treatment. Immunohistochemical analysis also showed that the 37-kDa adduct is predominantly localized in the pericentral region of the liver where CYP2E1 protein is mainly expressed. This staining disappeared in the pericentral region after YH439 treatment. The levels of alcohol dehydrogenase (ADH) and aldehyde dehydrogenase isozymes were unchanged after YH439 treatment. However, the level of the 37-kDa protein adduct positively correlated with the hepatic content of P4502E1, These data indicate that the 37-kDa adduct could be produced by CYP2E1-mediated ethanol metabolism in addition to the ADH-dependent formation. (C) 2000 Academic Press. C1 NIAAA, Lab Membrane Biochem & Biophys, Rockville, MD 20852 USA. Univ S Carolina, Dept Biol Sci, Columbia, SC 29208 USA. NE Ohio Med Sch, Dept Biochem, Rootstown, OH 44272 USA. RP Song, BJ (reprint author), NIAAA, Lab Membrane Biochem & Biophys, 12420 Parklawn Dr, Rockville, MD 20852 USA. NR 39 TC 23 Z9 23 U1 1 U2 3 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0003-9861 J9 ARCH BIOCHEM BIOPHYS JI Arch. Biochem. Biophys. PD DEC 1 PY 2000 VL 384 IS 1 BP 81 EP 87 DI 10.1006/abbi.2000.2119 PG 7 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 380JF UT WOS:000165697500010 PM 11147839 ER PT J AU Toro, JR Aksentijevich, I Hull, K Dean, J Kastner, DL AF Toro, JR Aksentijevich, I Hull, K Dean, J Kastner, DL TI Tumor necrosis factor receptor-associated periodic syndrome - A novel syndrome with cutaneous manifestations SO ARCHIVES OF DERMATOLOGY LA English DT Article; Proceedings Paper CT 61st Annual Meeting of the Society-for-Investigative-Dermatology CY MAY 10-14, 2000 CL CHICAGO, ILLINOIS SP Soc Investigat Dermatol ID FAMILIAL MEDITERRANEAN FEVER; RECURRENT HEREDITARY POLYSEROSITIS; HYPERIMMUNOGLOBULINEMIA-D SYNDROME; MULTISYSTEM INFLAMMATORY DISEASE; ENCODING MEVALONATE KINASE; HYPER-IGD SYNDROME; POLYARTERITIS-NODOSA; TNF RECEPTOR; CLINICAL SPECTRUM; CHROMOSOME 12P13 AB Background: Tumor necrosis factor receptor-associated periodic syndrome (TRAPS) is an inflammatory disorder characterized by prolonged episodes of periodic fever and localized inflammation and dominantly inherited mutations in TNFRSF1A, the gene encoding the 55-kDa tumor necrosis factor receptor. To our knowledge, the cutaneous pathologic characteristics of TRAPS have not been described previously. Objectives: To characterize the dermatologic manifestations of TRAPS by clinical, microscopic, and molecular methods, and to investigate its immunophenotype. Design, getting, and Patients: At the National Institutes of Health Clinical Center, Bethesda, Md, a tertiary care referral center, 25 patients with a clinical and molecular diagnosis of TRAPS were evaluated clinically and 10 biopsy specimens of lesional skin were examined by light microscopy and immunohistochemistry. Patients were screened for mutations in TNFRSF1A, the gene coding for the p55 tumor necrosis factor receptor. Main Outcome Measures: Clinical, light microscopic, and immunohistochemical features. Results: The skin eruption usually lasted 4 to 21 days (mean, 13 days). Of 25 patients, 21 (84%) presented with migratory erythematous macules and patches and 10 (40%) had edematous dermal plaques. Conjunctivitis, characterized by pain and redness and/or periorbital edema, was present in 11 patients (44%). Most patients had their first skin eruption during the first 2 years of life. All patients had fever associated with the skin eruption. Most patients had associated abdominal pain (22 [88%]) and myalgia (20 [80%]). Other symptoms included arthralgia (13 [52%]), pleuritic chest pain (10 [40%]), and headache (17 [68%]). Microscopic examination of 10 biopsy specimens of lesional skin showed a superficial and deep perivascular and interstitial infiltrate of lymphocytes and monocytes. None of the biopsy specimens showed multinucleated macrophages or granulomatous or leukocytoclastic vasculitis. The results of immunohistochemistry showed a perivascular infiltrate of CD3+, CD4+, CD8+, CD68+, CD79a-, and CD20- cells. All the mutations were missense mutations in exons 2 through 4 of TNFRSF1A, directly affecting the extracellular domain of the protein. Conclusions: TRAPS is characterized by a spectrum of dermatologic findings, including migratory patches, edematous plaques, periorbital edema, and/or conjunctivitis. TRAPS is characterized by a perivascular dermal infiltrate of lymphocytes and monocytes. C1 NCI, NIH, Bethesda, MD 20892 USA. NIAMSD, NIH, Bethesda, MD 20892 USA. RP Toro, JR (reprint author), NCI, NIH, Bldg 10,Room 12N-238,10 Ctr Dr,MSC 1908, Bethesda, MD 20892 USA. NR 74 TC 69 Z9 71 U1 0 U2 0 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0003-987X J9 ARCH DERMATOL JI Arch. Dermatol. PD DEC PY 2000 VL 136 IS 12 BP 1487 EP 1494 DI 10.1001/archderm.136.12.1487 PG 8 WC Dermatology SC Dermatology GA 383EA UT WOS:000165869100007 PM 11115159 ER PT J AU Toro, JR Finlay, D Dou, XG Zheng, SC LeBoit, PE Connolly, MK AF Toro, JR Finlay, D Dou, XG Zheng, SC LeBoit, PE Connolly, MK TI Detection of type 1 cytokines in discoid lupus erythematosus SO ARCHIVES OF DERMATOLOGY LA English DT Article; Proceedings Paper CT 58th Annual Meeting of the Society-for-Investigative-Dermatology CY APR 23-27, 1997 CL WASHINGTON, D.C. SP Soc Invest Dermatol ID TUMOR-NECROSIS-FACTOR; BLOOD MONONUCLEAR-CELLS; GENE-EXPRESSION; FACTOR-ALPHA; INTERFERON; INTERLEUKIN-6; MICE; RECEPTOR; PROFILE; SUBSETS AB Backgrounds: Although multiple studies suggest a dysregulated T-cell cytokine production in systemic lupus erythematosus, the cytokine profile in discoid lupus erythematosus (DLE) lesions is unknown. Objectives: To characterize the cytokine profile in DLE by immunohistochemical and molecular methods, and to investigate the role of cytokines in the pathogenesis of DLE. Design: Patients were evaluated clinically, and biopsy specimens of lesional skin were examined by light microscopy. Reverse transcriptase-polymerase chain reaction and immunohistochemical analysis were performed on 11 biopsy specimens. We investigated the presence of interleukin (IL) 2, interferon gamma (IFN-gamma), IL-4, tumor necrosis factor alpha, (TNF-alpha) and IL-1 beta messenger RNA (mRNA) in 8 biopsy specimens of DLE and compared it with 3 biopsy specimens of normal skin. Setting: Academic referral research hospital. Patients: Eight consecutive patients with a clinical and histologic diagnosis of DLE. Results: Localized DLE was found in 7 patients and widespread in 1. During the 4 years of the investigation, none of the patients developed systemic lupus erythematosus. We found significantly elevated levels of IL-2 and IFN-gamma mRNA in all 8 biopsy specimens of DLE; in contrast, no transcripts of IL-2 or IFN-gamma were detected in 3 biopsy specimens of normal skin (P<.01). Similarly, elevated levels of TNF- mRNA were detected in 8 DLE biopsy specimens of normal skin (P<.01). No IL-5 or IL-1 mRNA was detected in 8 biopsy specimens of DLE lesional skin and 3 biopsy specimens of normal patient skin. Immunohistochemical analysis showed increased staining for IL-2 and IFN-gamma receptors, while no detectable IL-4 receptor was found. No cytokine mRNA or cytokine receptor protein was detected in biopsy specimens of normal skin. Conclusions: These findings suggest that DLE is associated with type 1 cytokines characterized by the expression of IL-2 and IFN-gamma. Type 1 cytokines may be critical for induction, development, and maintenance of DLE. C1 Univ Calif San Francisco, Dept Dermatol, San Francisco, CA 94143 USA. RP Toro, JR (reprint author), NCI, NIH, Bldg 10,Room 12N-238,10 Ctr Dr,MSC 1908, Bethesda, MD 20892 USA. NR 23 TC 32 Z9 34 U1 0 U2 1 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0003-987X J9 ARCH DERMATOL JI Arch. Dermatol. PD DEC PY 2000 VL 136 IS 12 BP 1497 EP 1501 DI 10.1001/archderm.136.12.1497 PG 5 WC Dermatology SC Dermatology GA 383EA UT WOS:000165869100008 PM 11115160 ER PT J AU Watanabe, T Nakamura, K Wakugawa, M Kato, A Nagai, Y Shioda, T Iwamoto, A Tamaki, K AF Watanabe, T Nakamura, K Wakugawa, M Kato, A Nagai, Y Shioda, T Iwamoto, A Tamaki, K TI Antibodies to molluscum contagiosum virus in the general population and susceptible patients SO ARCHIVES OF DERMATOLOGY LA English DT Article ID CLINICAL LESIONS; POLYPEPTIDES; RESPONSES; LOCATION; SEQUENCE AB Background: Since many attempts to cultivate molluscum contagiosum virus (MCV) in vitro have been unsuccessful, it is difficult to prepare a large quantity of antigens. To assess the seroprevalence of antibodies against MCV in 508 subjects with or without clinical MCV infection, a truncated recombinant protein from open-reading frame MC133L was synthesized using Sendai virus expression system and applied to enzyme-linked immunosorbent assay as an antigen. Observations: Antibodies to MCV were present in 7 (58%) of 12 patients with molluscum contagiosum, 7 (6%) of 108 healthy controls, 7 (9%) of 76 with atopic dermatitis, and 7 (18%) of 39 patients with systemic lupus erythematosus, although no clinical MCV infection was observed in the latter 3 groups. Of 7 human immunodeficiency virus (HIV)-positive patients with molluscum contagiosum, 1 (14%) was antibody positive, compared with 5 (2%) of 266 HIV-positive patients without molluscum contagiosum. Serum samples from patients with atopic dermatitis and systemic lupus erythematosus showed a higher reactivity (P<.001) than those from healthy controls, while serum samples from HIV-positive subjects showed a lower reactivity (P<.001). Conclusion: The humoral immune response to MCV is usually confined to patients with molluscum contagiosum and may be affected by the immunological condition of the host. C1 Univ Tokyo, Fac Med, Dept Dermatol, Tokyo 113, Japan. Univ Tokyo, Inst Med Sci, Dept Viral Infect, Tokyo, Japan. Univ Tokyo, Inst Med Sci, Dept Infect Dis, Tokyo, Japan. RP Watanabe, T (reprint author), NCI, Dermatol Branch, Bldg 10,Room 12N238,10 Ctr Dr,MSC1908, Bethesda, MD 20892 USA. NR 19 TC 16 Z9 16 U1 0 U2 4 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0003-987X J9 ARCH DERMATOL JI Arch. Dermatol. PD DEC PY 2000 VL 136 IS 12 BP 1518 EP 1522 DI 10.1001/archderm.136.12.1518 PG 5 WC Dermatology SC Dermatology GA 383EA UT WOS:000165869100012 PM 11115164 ER PT J AU Cravchik, A Goldman, D AF Cravchik, A Goldman, D TI Neurochemical individuality - Genetic diversity among human dopamine and serotonin receptors and transporters SO ARCHIVES OF GENERAL PSYCHIATRY LA English DT Review ID SINGLE-NUCLEOTIDE POLYMORPHISMS; DEFICIT HYPERACTIVITY DISORDER; AMINO-ACID SUBSTITUTIONS; DRD2 VARIANT SER311CYS; HUMAN 5-HT1A RECEPTOR; MICE LACKING; NOVELTY SEEKING; MUTANT MICE; ANOREXIA-NERVOSA; D2 RECEPTOR AB Behavioral variation in human beings encompasses wide differences in personality and susceptibility to psychiatric illness arising from both genotype and experience. Longlasting behavioral differences generally have heritabilities of 30% or more, and such inheritance is ultimately attributable to functional variants of genes programming brain development and function. The sequencing of the human genome is revealing a pattern of gene sequence variation. The ability of sequence variants to affect neural function either alone or in concert may reveal effects of behavioral selection on the human genome over evolutionary time frames. Dopamine and serotonin are phylogenetically ancient neurotransmitters intrinsic to brain function and behavior. Dopamine and serotonin receptor and transporter genes have been an early focus for efforts to identify and functionally characterize sequence variation. The purpose of this article is to present a preview of a developing new perspective in human behavior: the genetic variation of the brain or neurochemical individuality. C1 Celera Genom, Rockville, MD 20850 USA. NIAAA, Neurogenet Lab, NIH, Bethesda, MD USA. RP Cravchik, A (reprint author), Celera Genom, 45 W Gude Dr, Rockville, MD 20850 USA. RI Goldman, David/F-9772-2010 OI Goldman, David/0000-0002-1724-5405 NR 116 TC 62 Z9 66 U1 6 U2 14 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0003-990X J9 ARCH GEN PSYCHIAT JI Arch. Gen. Psychiatry PD DEC PY 2000 VL 57 IS 12 BP 1105 EP 1114 DI 10.1001/archpsyc.57.12.1105 PG 10 WC Psychiatry SC Psychiatry GA 379XF UT WOS:000165667900001 PM 11115324 ER PT J AU Cohen, F Cole, SR Beck, RW Blair, RC Moke, PS Kip, KE Wilhelmus, KR Dawson, CR Barron, BA Jones, DB Kaufman, HE Stulting, RD Sugar, J Cohen, EJ Hyndiuk, RA Asbell, PA Kurinij, N Edwards, K Massare, SJ McGovern, MS Williams, D Dragon, DM Graul, EE Insler, MS Kaufman, HE Lacoste, AD McCaa, CS Wagner, NJ Yokubaitis, JA Wilhelmus, KR Todaro, LA Woodside, SJ Bowman, CB Chodosh, J Goosey, JD Jones, DB Kirkland, C Lehmann, RP Matoba, AY Smith, SL Wolf, TC Yee, RW Cohen, F Dawson, CR Banuvar, S Osaki, SY Barth, GP Biswell, R Cunningham, E DeMartini, DR Gritz, D Hodge, W Holsclaw, DS Hwang, DG Knox, CM Lietman, T Margolis, TP Schwab, IR Schwartz, L Sherman, M Silverstein, B Vastine, DW Volpicelli, M Whitcher, JP Wilson, S Wong, I Stulting, RD DuBois, LG Baldassare, R Chopra, H Croll, SD DiIorio, RC Gussler, JR Hamilton, SM John, GR McCann, JW Meyer, JC Mitchell, PG Palay, DA Ramirez, RJ Reed, RE Serros, RN Taub, LR Thompson, KP Walter, KA Sugar, J Rodiek, R Dennis, R Feder, RS Hennessey, MJ Lubeck, DM McLeod, SD Meisler, D Morimoto, DD Morimoto, PK Rubenstein, JB Tessler, HH Hyndiuk, RA Samson, C Barney, NP Brightbill, FS DeCarlo, JD Fogel, ES Gainey, SP Koenig, SB Kontra, DJ Krebs, DB Lewellen, DR Patalano, SM Rice, PR Sanderson, MC Wienkers, KP Yeomans, MM Cohen, EJ Marshall, SC Rodriguez, IM Phipps, P Bailey, RJ Fung, KL Hannush, SB Ingraham, HI Kesselring, JJ Kowal, VO Laibson, PR Martin, NF Orlin, SO Rapuano, CR Rubinefeld, RS Sulewski, M Asbell, PA Justin, N Arroyo, M Epstein, S Barker, BA Brocks, RE D'Aversa, G Dunn, MJ Gorman, BD Leopold, MR Schwartz, WJ Udell, IJ Beck, RW Moke, PS Blair, RC Cole, SR Kip, K Gillespie, HA Lester, LA Mhamdi, ML Gal, RL Tan, ES Hauck, WW Gee, L Bangdiwala, SI Barlow, WE Chandler, JW Lemp, MA Nesburn, AB Patrick, DL Sutphin, JE Watson, SB AF Cohen, F Cole, SR Beck, RW Blair, RC Moke, PS Kip, KE Wilhelmus, KR Dawson, CR Barron, BA Jones, DB Kaufman, HE Stulting, RD Sugar, J Cohen, EJ Hyndiuk, RA Asbell, PA Kurinij, N Edwards, K Massare, SJ McGovern, MS Williams, D Dragon, DM Graul, EE Insler, MS Kaufman, HE Lacoste, AD McCaa, CS Wagner, NJ Yokubaitis, JA Wilhelmus, KR Todaro, LA Woodside, SJ Bowman, CB Chodosh, J Goosey, JD Jones, DB Kirkland, C Lehmann, RP Matoba, AY Smith, SL Wolf, TC Yee, RW Cohen, F Dawson, CR Banuvar, S Osaki, SY Barth, GP Biswell, R Cunningham, E DeMartini, DR Gritz, D Hodge, W Holsclaw, DS Hwang, DG Knox, CM Lietman, T Margolis, TP Schwab, IR Schwartz, L Sherman, M Silverstein, B Vastine, DW Volpicelli, M Whitcher, JP Wilson, S Wong, I Stulting, RD DuBois, LG Baldassare, R Chopra, H Croll, SD DiIorio, RC Gussler, JR Hamilton, SM John, GR McCann, JW Meyer, JC Mitchell, PG Palay, DA Ramirez, RJ Reed, RE Serros, RN Taub, LR Thompson, KP Walter, KA Sugar, J Rodiek, R Dennis, R Feder, RS Hennessey, MJ Lubeck, DM McLeod, SD Meisler, D Morimoto, DD Morimoto, PK Rubenstein, JB Tessler, HH Hyndiuk, RA Samson, C Barney, NP Brightbill, FS DeCarlo, JD Fogel, ES Gainey, SP Koenig, SB Kontra, DJ Krebs, DB Lewellen, DR Patalano, SM Rice, PR Sanderson, MC Wienkers, KP Yeomans, MM Cohen, EJ Marshall, SC Rodriguez, IM Phipps, P Bailey, RJ Fung, KL Hannush, SB Ingraham, HI Kesselring, JJ Kowal, VO Laibson, PR Martin, NF Orlin, SO Rapuano, CR Rubinefeld, RS Sulewski, M Asbell, PA Justin, N Arroyo, M Epstein, S Barker, BA Brocks, RE D'Aversa, G Dunn, MJ Gorman, BD Leopold, MR Schwartz, WJ Udell, IJ Beck, RW Moke, PS Blair, RC Cole, SR Kip, K Gillespie, HA Lester, LA Mhamdi, ML Gal, RL Tan, ES Hauck, WW Gee, L Bangdiwala, SI Barlow, WE Chandler, JW Lemp, MA Nesburn, AB Patrick, DL Sutphin, JE Watson, SB CA Herpetic Eye Dis Study Grp TI Psychological stress and other potential triggers for recurrences of herpes simplex virus eye infections SO ARCHIVES OF OPHTHALMOLOGY LA English DT Article ID SOCIAL DESIRABILITY SCALE; GENITAL HERPES; PSYCHOSOCIAL FACTORS; CLINICAL-FEATURES; PREDICTORS; LABIALIS; EPIDEMIOLOGY; REACTIVATION; PREVENTION; REGRESSION AB Objective: To assess psychological stress and other factors as possible triggers of ocular herpes simplex virus (HSV) recurrences. Design: A prospective cohort study nested in a randomized, placebo-controlled, clinical trial. Setting: Fifty-eight community-based or university sites. Participants: Immunocompetent adults (N = 308), aged 18 years or older, with a documented history of ocular HSV disease in the prior year and observed for up to 15 months. Exposure Variables: Psychological stress, systemic infection, sunlight exposure, menstrual period, contact lens wear, and eye injury recorded on a weekly log. The exposure period was considered to be the week before symptomatic onset of a recurrence. Main Outcome Measure: The first documented recurrence of ocular HSV disease, with exclusion of cases in which the exposure week log was completed late after the onset of symptoms. Results: Thirty-three participants experienced a study outcome meeting these criteria. Higher levels of psychological stress were not associated with an increased risk of recurrence (rate ratio, 0.58; 95%;, confidence interval, 0.32-1.05; P=.07). No association was found between any of the other exposure variables and recurrence. When an analysis was performed including only the recurrences (n=26) for which the exposure week log was completed late and after symptom onset, there was a clear indication of retrospective overreporting of high stress (P =.03) and systemic infection (p=.01). Not excluding these cases could have produced incorrect conclusions due to recall bias. Conclusions: Psychological stress does not appear to be a trigger of recurrences of ocular HSV disease. If not accounted for, recall bias can substantially overestimate the importance of factors that do not have a causal association with HSV infection. C1 Jaeb Ctr Hlth Res Inc, Tampa, FL 33613 USA. Univ Calif San Francisco, San Francisco, CA 94143 USA. NEI, NIH, Bethesda, MD 20892 USA. Louisiana State Univ, Med Ctr, Ctr Eye, New Orleans, LA 70112 USA. Baylor Coll Med, Cullen Eye Inst, Houston, TX 77030 USA. Univ Calif San Francisco, Francis I Proctor Fdn, San Francisco, CA 94143 USA. Emory Univ, Ctr Eye, Atlanta, GA 30322 USA. Univ Illinois, Ctr Eye, Chicago, IL 60680 USA. Med Coll Wisconsin, Inst Eye, Milwaukee, WI 53226 USA. Wills Eye Hosp & Res Inst, Philadelphia, PA 19107 USA. CUNY, Mt Sinai Med Ctr, New York, NY 10021 USA. RP Beck, RW (reprint author), Jaeb Ctr Hlth Res Inc, 3010 E 138th Ave,Suite 9, Tampa, FL 33613 USA. EM rbeck@jaeb.org NR 45 TC 25 Z9 25 U1 2 U2 5 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60654-0946 USA SN 0003-9950 EI 1538-3601 J9 ARCH OPHTHALMOL-CHIC JI Arch. Ophthalmol. PD DEC PY 2000 VL 118 IS 12 BP 1617 EP 1625 PG 9 WC Ophthalmology SC Ophthalmology GA 380ZX UT WOS:000165738800001 ER PT J AU Dickerman, RD Douglas, JA East, JW AF Dickerman, RD Douglas, JA East, JW TI Bilateral median neuropathy and growth hormone use: A case report SO ARCHIVES OF PHYSICAL MEDICINE AND REHABILITATION LA English DT Article DE peripheral neuropathies; carpal tunnel syndrome; growth hormones, recombinant; case report; rehabilitation ID MEN; THERAPY; OLDER AB A male elite bodybuilder suffered bilateral median nerve neuropathy during a self-administered course of growth hormone (GH). Nerve conduction velocities revealed bilateral median neuropathy consistent with carpal tunnel syndrome (CTS). This is the first case of GH-induced CTS occurring in an athlete. Contrary to earlier studies, this report demonstrates that GH-induced CTS is not an age-related phenomenon and alerts physicians to include GH abuse as a possible etiology of median neuropathy in athletes. C1 NIH, Surg Neurol Branch, Bethesda, MD 20892 USA. Univ N Texas, Hlth Sci Ctr, Dept Surg, Ft Worth, TX USA. Univ Texas, SW Med Ctr, Dept Phys Med & Rehabil, Dallas, TX USA. RP Dickerman, RD (reprint author), NIH, Surg Neurol Branch, 10 Ctr Dr,Bldg 10,Rm 5D37, Bethesda, MD 20892 USA. NR 12 TC 7 Z9 7 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0003-9993 J9 ARCH PHYS MED REHAB JI Arch. Phys. Med. Rehabil. PD DEC PY 2000 VL 81 IS 12 BP 1594 EP 1595 DI 10.1053/1pmr.2000.7159 PG 2 WC Rehabilitation; Sport Sciences SC Rehabilitation; Sport Sciences GA 380JU UT WOS:000165698800010 PM 11128896 ER PT J AU Iba, MM Fung, J Gonzalez, FJ AF Iba, MM Fung, J Gonzalez, FJ TI Functional Cyp2e1 is required for substantial in vivo formation of 2,5-hexanedione from n-hexane in the mouse SO ARCHIVES OF TOXICOLOGY LA English DT Article DE Cyp2e1-knockout mouse; n-Hexane; biotransformation; 2,5-hexanedione ID METHYL ETHYL KETONE; NORMAL-BUTYL KETONE; RAT-LIVER MICROSOMES; CYTOCHROME-P-450; METABOLITES; HYDROXYLATION; PARKINSONISM; MICE; 4,5-DIHYDROXY-2-HEXANONE; NEUROTOXICITY AB Neurotoxicity of n-hexane is mediated by its metabolite 2,5-hexanedione (2,5-HD). Cytochrome P4502E1 (CYP2E1) has been suggested but not shown to be involved in the formation of the metabolite. An objective of the current study was to assess the essentiality of CYP2E1 for in vivo 2,5-HD formation from n-hexane. This was accomplished by comparing urinary levels of the gamma -diketone in n-hexane-treated mice in which the Cyp2e1 gene has been deleted(Cyp2e1(-/-)) with that in n-hexane-treated wild-type (Cyp2e1(+/+)) mice. 2,5-HD was detectable not as the free compound but as further metabolites, at levels that were comparable in both strains of mice, following a daily 200 mg/kg i.p. dose of the alkane for 10 days. Continued daily n-hexane treatment resulted in increased urinary levels of 2,5-HD metabolites in Cyp2e1(+/+) but not in Cyp2e1(-/-) mice. Only in Cyp2e1(+/+) mice and only on day 21 of n-hexane treatment was a trace level of unchanged 2,5-HD detected. 3-Hexanol was the only other n-hexane metabolite detected in the mice but its concentration was higher in Cyp2e1(-/-) than in Cyp2e1(+/+) mice. In n-hexane-treated rats, in contrast to mice, multiple metabolites of the alkane, including unchanged 2,5-HD, were detected. The results indicate that substantial in vivo formation of 2,5-HD from n-hexane in the mouse requires CYP2E1, and suggest that further detoxification of the metabolite may be very efficient in this species. C1 Rutgers State Univ, Dept Pharmacol & Toxicol, EOHSI, Piscataway, NJ 08854 USA. NCI, Mol Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. RP Iba, MM (reprint author), Rutgers State Univ, Dept Pharmacol & Toxicol, EOHSI, 170 Frelinghuysen Rd, Piscataway, NJ 08854 USA. FU NIEHS NIH HHS [ES06414, ES05221] NR 37 TC 10 Z9 16 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0340-5761 J9 ARCH TOXICOL JI Arch. Toxicol. PD DEC PY 2000 VL 74 IS 10 BP 582 EP 586 DI 10.1007/s002040000182 PG 5 WC Toxicology SC Toxicology GA 385LZ UT WOS:000166007100003 PM 11201664 ER PT J AU Imperatore, G Knowler, WC Pettitt, DJ Kobes, S Fuller, JH Bennett, PH Hanson, RL AF Imperatore, G Knowler, WC Pettitt, DJ Kobes, S Fuller, JH Bennett, PH Hanson, RL TI A locus influencing total serum cholesterol on chromosome 19p - Results from an autosomal genomic scan of serum lipid concentrations in Pima Indians SO ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY LA English DT Article DE cholesterol; HDL cholesterol; linkage; genetics; triglycerides ID DENSITY-LIPOPROTEIN CHOLESTEROL; QUANTITATIVE-TRAIT LOCI; LDL RECEPTOR GENE; BODY-MASS INDEX; ENZYMATIC DETERMINATION; LINKAGE ANALYSIS; FAMILIES; POLYMORPHISMS; SEGREGATION; POPULATION AB A genome-wide linkage study was analyzed to identify loci that influence serum lipid concentrations in Pima Indians, Linkage analyses were conducted for total cholesterol measured in 998 siblings from 292 nuclear families, for total triglycerides in 547 siblings from 188 families, and for high density lipoprotein (HDL) cholesterol in 590 siblings from 201 families. Genotypes were generated for 516 autosomal microsatellite markers. Multipoint variance components methods were used to assess linkage. The strongest evidence for linkage with total cholesterol was on chromosome 19p (lod score 3.89), in the vicinity of the marker D19S1034, which is near the low density lipoprotein receptor gene. The strongest evidence for linkage with HDL cholesterol was on chromosome 3q (lod score 2.64) near D3S3053, For triglycerides, the strongest evidence for linkage was on chromosome 2p near D2S1788 (lod score 1.70) and on chromosome 3p near D3S2406 (lod score 1.77). This genomic scan provides evidence for a locus influencing total cholesterol concentration on chromosome 19p. It also suggests a locus influencing HDL cholesterol on chromosome 3q. C1 NIDDK, Diabet & Arthrit Epidemiol Sect, Phoenix Epidemiol & Clin Res Branch, NIH, Phoenix, AZ 85014 USA. RP Hanson, RL (reprint author), NIDDK, Diabet & Arthrit Epidemiol Sect, Phoenix Epidemiol & Clin Res Branch, NIH, 1550 E Indian Sch Rd, Phoenix, AZ 85014 USA. RI Hanson, Robert/O-3238-2015 OI Hanson, Robert/0000-0002-4252-7068 NR 36 TC 59 Z9 61 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1079-5642 J9 ARTERIOSCL THROM VAS JI Arterioscler. Thromb. Vasc. Biol. PD DEC PY 2000 VL 20 IS 12 BP 2651 EP 2656 PG 6 WC Hematology; Peripheral Vascular Disease SC Hematology; Cardiovascular System & Cardiology GA 383EJ UT WOS:000165869900031 PM 11116067 ER PT J AU Sneller, MC AF Sneller, MC TI Cystitis, bladder cancer, and myelodysplasia in patients with Wegener's granulomatosis: comment on the article by Reinhold-Keller et al SO ARTHRITIS AND RHEUMATISM LA English DT Letter C1 NIAID, Bethesda, MD 20892 USA. RP Sneller, MC (reprint author), NIAID, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 3 TC 12 Z9 12 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0004-3591 J9 ARTHRITIS RHEUM JI Arthritis Rheum. PD DEC PY 2000 VL 43 IS 12 BP 2853 EP 2854 DI 10.1002/1529-0131(200012)43:12<2853::AID-ANR30>3.0.CO;2-G PG 2 WC Rheumatology SC Rheumatology GA 383ET UT WOS:000165870700029 PM 11145048 ER PT J AU Costa, PT McCrae, RR AF Costa, PT McCrae, RR TI Overview: Innovations in assessment using the revised NEO Personality Inventory SO ASSESSMENT LA English DT Article; Proceedings Paper CT 108th Annual Meeting of the American-Psychological-Association CY AUG 04-08, 2000 CL WASHINGTON, D.C. SP Amer Psychol Assoc ID VALIDITY SCALES; SELF-REPORTS; VALIDATION; ROTATION; RATINGS C1 NIA, NIH, Baltimore, MD 21224 USA. RP Costa, PT (reprint author), Ctr Gerontol Res, Lab Personal & Cognit, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. OI Costa, Paul/0000-0003-4375-1712 NR 19 TC 21 Z9 22 U1 3 U2 12 PU SAGE PUBLICATIONS INC PI THOUSAND OAKS PA 2455 TELLER RD, THOUSAND OAKS, CA 91320 USA SN 1073-1911 J9 ASSESSMENT JI Assessment PD DEC PY 2000 VL 7 IS 4 BP 325 EP 327 DI 10.1177/107319110000700402 PG 3 WC Psychology, Clinical SC Psychology GA 531YU UT WOS:000174445500002 PM 11151960 ER PT J AU Costa, PT Herbst, JH McCrae, RR Siegler, IC AF Costa, PT Herbst, JH McCrae, RR Siegler, IC TI Personality at midlife: Stability, intrinsic maturation, and response to life events SO ASSESSMENT LA English DT Article; Proceedings Paper CT Annual Meeting of the American-Psychological-Association CY AUG 18-24, 1999 CL BOSTON, MASSACHUSETTS SP Amer Psychol Assoc DE aging; life events; longitudinal; midlife; traits ID AGE; WOMEN; SPAN; INVENTORY; CULTURES; RATINGS; TRAITS AB Although developmental theories and popular accounts suggest that midlife is a time of turmoil and change, longitudinal studies of personality traits have generally found stability of rank order and little or no change in mean levels. Using data from 2,274 men and women in their 40s retested after 6 to 9 years, the present study examined two hypotheses: (a) that retest correlations should be no higher than about .60 and (b) that there should be small decreases in Neuroticism, Extraversion, and Openness, and small increases in Agreeableness and Conscientiousness. The study also explored the effects of recalled life events on subsequent personality scores. Results did not support the first hypothesis; uncorrected retest correlations uniformly exceeded .60. This was true for all personality traits, including facets of Agreeableness and Conscientiousness not previously included in longitudinal studies. The hypothesized decreases in Neuroticism, Extraversion, and Openness were found, but Conscientiousness showed a small decrease instead of the predicted increase. Life events in general showed very little influence on the levels of personality traits, although some effects were seen for changes in job and marital status that warrant further research. C1 NIA, NIH, Intramural Res Program, Personal Stress & Coping Sect,Lab Persona & Cogni, Baltimore, MD 21224 USA. Duke Univ, Med Ctr, Behav Med Res Ctr, Durham, NC USA. RP Costa, PT (reprint author), NIA, NIH, Intramural Res Program, Personal Stress & Coping Sect,Lab Persona & Cogni, Baltimore, MD 21224 USA. OI Costa, Paul/0000-0003-4375-1712 FU NHLBI NIH HHS [HL36587, HL55356]; NIA NIH HHS [AG12458] NR 37 TC 172 Z9 175 U1 2 U2 19 PU SAGE PUBLICATIONS INC PI THOUSAND OAKS PA 2455 TELLER RD, THOUSAND OAKS, CA 91320 USA SN 1073-1911 J9 ASSESSMENT JI Assessment PD DEC PY 2000 VL 7 IS 4 BP 365 EP 378 DI 10.1177/107319110000700405 PG 14 WC Psychology, Clinical SC Psychology GA 531YU UT WOS:000174445500005 PM 11151962 ER PT J AU Herbst, JH McCrae, RR Costa, PT Feaganes, JR Siegler, IC AF Herbst, JH McCrae, RR Costa, PT Feaganes, JR Siegler, IC TI Self-perceptions of stability and change in personality at midlife: The UNC Alumni Heart Study SO ASSESSMENT LA English DT Article; Proceedings Paper CT Annual Meeting of the American-Psychological-Association CY AUG 18-24, 1999 CL BOSTON, MASSACHUSETTS SP Amer Psychol Assoc DE personality; stability; perceptions; longitudinal; midlife ID ADULT LIFE-SPAN; SCALES AB The finding of personality stability in adulthood may be counterintuitive to people who perceive a great deal of change in their own personality. The purpose of this study is to determine whether self-reported perceived changes in personality are associated with actual changes based on a 6- to 9-year follow-up of 2,242 middle-aged male and female participants of the UNC Alumni Heart Study (UNCAHS). Respondents completed the Revised NEO Personality Inventory on two occasions and were asked to reflect back over a 6-year period and assess changes in their personality. The majority of respondents (n = 1,177; 52.5%) reported they had "stayed the same," while 863 (38.5%) reported they had "changed a little" and 202 (9%) reported they had "changed a good deal." Coefficients of personality profile agreement computed to evaluate global personality change for the three perceived change groups were essentially equivalent. Further, directional analyses of domain-specific changes in personality showed that perceived changes were weak predictors of residual gain scores. In an absolute sense, perceptions of stability or change were discordant in 8 of 15 (53%) comparisons. Self-perceptions of change are not an adequate substitute for objective assessments. C1 NIA, NIH, Lab Personal & Cognit, Intramural Res Program, Baltimore, MD 21224 USA. Duke Univ, Med Ctr, Behav Med Res Ctr, Durham, NC USA. RP Costa, PT (reprint author), NIA, NIH, Lab Personal & Cognit, Intramural Res Program, Baltimore, MD 21224 USA. OI Costa, Paul/0000-0003-4375-1712 FU NHLBI NIH HHS [HL36587, HL55356]; NIA NIH HHS [AG12458] NR 24 TC 10 Z9 13 U1 0 U2 3 PU SAGE PUBLICATIONS INC PI THOUSAND OAKS PA 2455 TELLER RD, THOUSAND OAKS, CA 91320 USA SN 1073-1911 J9 ASSESSMENT JI Assessment PD DEC PY 2000 VL 7 IS 4 BP 379 EP 388 DI 10.1177/107319110000700406 PG 10 WC Psychology, Clinical SC Psychology GA 531YU UT WOS:000174445500006 PM 11151963 ER PT J AU Kronenberg, F Pereira, MA Schmitz, MKH Arnett, DK Evenson, KR Crapo, RO Jensen, RL Burke, GL Sholinsky, P Ellison, RC Hunt, SC AF Kronenberg, F Pereira, MA Schmitz, MKH Arnett, DK Evenson, KR Crapo, RO Jensen, RL Burke, GL Sholinsky, P Ellison, RC Hunt, SC TI Influence of leisure time physical activity and television watching on atherosclerosis risk factors in the NHLBI Family Heart Study SO ATHEROSCLEROSIS LA English DT Article DE physical activity; TV watching; atherosclerosis risk factors; obesity; lipids; lipoproteins ID DENSITY-LIPOPROTEIN CHOLESTEROL; UNITED-STATES; LIFE-STYLE; CARDIOVASCULAR HEALTH; GLUCOSE-TOLERANCE; RANDOMIZED-TRIAL; NATIONAL-HEALTH; WEIGHT CHANGE; YOUNG-ADULTS; OBESITY AB Physical activity favorably influences atherosclerosis risk factors but only a few studies in adults considered the time watching television (TV) as a measure of physical inactivity. We therefore determined in a population-based sample of 1778 subjects from the NHLBI Family Heart Study (FHS) whether leisure time physical activity and TV watching have independent or interactive associations with cardiovascular disease risk factors and carotid artery intima-media wall thickness (IMT). Subjects were free from diabetes mellitus and clinically-ascertained coronary artery disease and did not take lipid-lowering or antihypertensive drugs. Only 0.7 and 1.3% of the variance in leisure time physical activity in women and men, respectively, was explained by the amount of TV watching. Leisure time physical activity had a clearly favorable, and TV watching an unfavorable association with anthropometric measurements (BMI (body mass index), waist girth, waist-hip ratio, subscapular and triceps skinfold thickness). The odds ratio (95% CI) of being overweight was 0.41 (0.28-0.62) in women and 0.69 (0.46-1.03) in men in the highest quartile of leisure time physical activity compared to the lowest quartile. The odds ratio increased for increasing quartiles of TV watching to 2.12 (1.45-3.10) in women and 1.61 (1.07-2.43) in men. Watching TV only 1 h per day in women with a BMI of 30 kg/m(2) and doing about 75 min of moderate exercise per week was associated with a BMI 1.8 kg/m(2) lower than in women watching TV 3 h per day and doing the same amount of exercise. Those with twice the amount of moderate exercise and watching TV 1 h per day had a BMI 0.45 kg/m(2) lower. Furthermore, leisure time physical activity was negatively associated with concentrations of triglycerides and positively with HDL cholesterol in both genders. TV watching was significantly positively associated with triglycerides and slightly negatively with HDL cholesterol in men. The observed associations of leisure time physical activity and TV watching with atherosclerosis risk factors were independent from each other. Finally, we analyzed the relation between leisure time physical activity, TV watching and the degree of IMT of the carotid arteries. Neither of these two measures was significantly associated with IMT. In summary, TV watching, in addition to leisure time physical activity, shows an independent association with obesity-related anthropometric measurements, HDL and triglycerides. Decreasing the amount of TV watching might be effective as a first step in reducing atherosclerosis risk factors, especially overweight. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved. C1 Univ Utah, Salt Lake City, UT 84112 USA. Childrens Hosp, Div Endocrinol, Boston, MA 02115 USA. Univ Minnesota, Div Epidemiol, Minneapolis, MN 55455 USA. Univ N Carolina, Dept Epidemiol, Chapel Hill, NC 27515 USA. LDS Hosp, Pulm Div, Salt Lake City, UT USA. Wake Forest Univ, Sch Med, Dept Publ Hlth Sci, Winston Salem, NC 27109 USA. NHLBI, Div Epidemiol & Clin Applicat, Bethesda, MD 20892 USA. Boston Univ, Sch Med, Sect Prevent Med & Epidemiol, Boston, MA 02118 USA. RP Kronenberg, F (reprint author), Univ Innsbruck, Inst Med Biol & Human Genet, Schopfstr 41, A-6020 Innsbruck, Austria. EM florian.kronenberg@uibk.ac.at RI Kronenberg, Florian/B-1736-2008 OI Kronenberg, Florian/0000-0003-2229-1120 FU NHLBI NIH HHS [UO1-HL-56563, UO1-HL-56564, UO1-HL-56565] NR 53 TC 120 Z9 121 U1 2 U2 4 PU ELSEVIER IRELAND LTD PI CLARE PA ELSEVIER HOUSE, BROOKVALE PLAZA, EAST PARK SHANNON, CO, CLARE, 00000, IRELAND SN 0021-9150 J9 ATHEROSCLEROSIS JI Atherosclerosis PD DEC PY 2000 VL 153 IS 2 BP 433 EP 443 DI 10.1016/S0021-9150(00)00426-3 PG 11 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 382YE UT WOS:000165854300019 PM 11164433 ER PT J AU Siahpush, M Singh, GK AF Siahpush, M Singh, GK TI A multivariate analysis of the association between social class of origin and current social class with self-rated general health and psychological health among 16-year-old Australians SO AUSTRALIAN AND NEW ZEALAND JOURNAL OF MEDICINE LA English DT Article DE social class and health in early youth; adolescent health; health equalisation hypothesis ID YOUNG MEN; YOUTH; INEQUALITIES; UNEMPLOYMENT; QUESTIONNAIRE; DIFFERENTIALS; DISTURBANCE; ADOLESCENCE; COMMUNITY; MORBIDITY AB Background: A recent review of international literature concludes that there is a relative absence of social class differentials in health in early youth. There is an absence of Australian studies on the effect of social class on the health of this age group. Aims: To examine the association between social class and health among 16-year-old Australians. Methods: The data on 1048 16-year-olds came from the fifth wave (1993) of the Australian Youth Survey conducted by the former Department of Employment, Education and Training. Outcome measures were self-rated general health and psychological health (GHQ-12). Binary logistic regression was used to analyse data. Results: Neither social class of origin nor current social class was associated with self-rated general health or psychological health. Conclusion: The argument that social class inequalities in health exist in childhood, disappear during early youth, and reappear later appears to hold ground within the Australian context. C1 Anti Canc Council Victoria, VicHlth Ctr Tobacco Control, Carlton, Vic 3053, Australia. NCI, NIH, Bethesda, MD 20892 USA. RP Siahpush, M (reprint author), Anti Canc Council Victoria, VicHlth Ctr Tobacco Control, 100 Drummond St, Carlton, Vic 3053, Australia. NR 39 TC 15 Z9 15 U1 4 U2 6 PU ADIS PRESS AUSTRALASIA P/L PI BALGOWLAH PA P.O. BOX 132, BALGOWLAH, NSW 2093, AUSTRALIA SN 0004-8291 J9 AUST NZ J MED JI Aust. N. Z. J. Med. PD DEC PY 2000 VL 30 IS 6 BP 653 EP 659 DI 10.1111/j.1445-5994.2000.tb04359.x PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 391NX UT WOS:000166363000003 PM 11198572 ER PT J AU Lee, JH Klein, HG AF Lee, JH Klein, HG TI From leukocyte reduction to leukocyte transfusion: the immunological effects of transfused leukocytes SO BEST PRACTICE & RESEARCH CLINICAL HAEMATOLOGY LA English DT Review DE leukocyte reduction; leukocyte transfusion; immunomodulation; transfusion-associated graft-versus-host disease; donor lymphocyte infusion; graft-versus-leukaemia effect ID BONE-MARROW TRANSPLANTATION; CHRONIC MYELOID-LEUKEMIA; GRAFT-VERSUS-LEUKEMIA; CHRONIC MYELOGENOUS LEUKEMIA; HUMAN-IMMUNODEFICIENCY-VIRUS; T-CELL DEPLETION; MINOR HISTOCOMPATIBILITY ANTIGEN; RECURRENT SPONTANEOUS-ABORTIONS; PERIPHERAL-BLOOD LYMPHOCYTES; ACTIVATED KILLER CELLS AB In transfusion medicine, mononuclear leukocytes have been studied more often as contaminants of red blood cells or platelets responsible for adverse transfusion outcomes than as therapeutic cells; leukocyte transfusion has been effective in augmenting recipient immunity only in limited clinical situations. Studies in leukocyte reduction and leukocyte transfusion have progressed separately as if the leukocytes' adverse and therapeutic effects result from different immunological mechanisms. With growing clinical experience, however, it is increasingly clear that some adverse immune effects may be exploited for therapeutic benefit. Advances in clinical immunology, understanding of the variety of cells and functions in the leukocyte fraction of blood, and blood component preparation technology may lead to new ways of deriving immunological benefit from transfused blood leukocytes while minimizing their adverse effects. This chapter reviews the current uses of leukocyte reduction and mononuclear leukocyte transfusion, with an emphasis on the relationship between transfusion-associated graft-versus-host disease and donor lymphocyte infusion in controlling relapsed leukaemias. C1 US FDA, Ctr Biol Evaluat & Res, Off Blood Res & Review, Div Blood Applicat,Blood & Plasma Branch, Rockville, MD 20852 USA. NIH, Warren G Magnuson Clin Ctr, Div Transfus Med, Bethesda, MD 20892 USA. RP Lee, JH (reprint author), US FDA, Ctr Biol Evaluat & Res, Off Blood Res & Review, Div Blood Applicat,Blood & Plasma Branch, HFM-375,1401 Rockville Pike, Rockville, MD 20852 USA. NR 105 TC 7 Z9 7 U1 0 U2 0 PU BAILLIERE TINDALL PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 1521-6926 J9 BEST PRACT RES CL HA JI Best Pract. Res. Clin. Haematol. PD DEC PY 2000 VL 13 IS 4 BP 585 EP 600 DI 10.1053/beha.2000.0101 PG 16 WC Hematology SC Hematology GA 383JC UT WOS:000165878500007 PM 11102278 ER PT J AU Brown, MR Blanchette, JO Kohn, EC AF Brown, MR Blanchette, JO Kohn, EC TI Angiogenesis in ovarian cancer SO BEST PRACTICE & RESEARCH IN CLINICAL OBSTETRICS & GYNAECOLOGY LA English DT Review DE angiogenesis; ovarian cancer; folliculogenesis; vascular endothelial growth factor; basic fibroblast growth factor; anti-angiogenic therapy ID ENDOTHELIAL GROWTH-FACTOR; VASCULAR-PERMEABILITY FACTOR; TUMOR-SUPPRESSOR GENE; HUMAN BREAST-CANCER; CARCINOMA CELLS; CORPUS-LUTEUM; IN-VIVO; ELEVATED LEVELS; UP-REGULATION; FACTOR VEGF AB Angiogenesis, the formation of new vessels from pre-existing vasculature, is critical to ascites development and metastasis in ovarian cancer. Many growth factors important to ovarian cancer invasion are also prominent in its associated angiogenesis. Deregulation of normal angiogenic processes occurs with the cancer's acquisition of the ability to secrete proangiogenic factors. The local imbalance of endogenous angio-stimulators and angio-inhibitors promotes vascularization and vascular leak. Assessment of these pro-angiogenic growth factors and enumeration of tumour-associated microvessels have been shown to be prognosticators of ovarian cancer outcome, and may also be surrogates of ovarian cancer tumour burden and/or ascites formation. The process of angiogenesis has been targeted for therapeutics development. Ovarian cancer is a primary cancer against which these new agents are being tested. Thus, further understanding of the molecular and cell biology of angiogenesis in the context of ovarian cancer offers important directions for estimation of patient outcome and for patient treatment. C1 NCI, Mol Signaling Sect, Pathol Lab, Bethesda, MD 20892 USA. RP Kohn, EC (reprint author), NCI, Mol Signaling Sect, Pathol Lab, Bldg 10,Room 2A33,10 Ctr Dr, Bethesda, MD 20892 USA. NR 108 TC 35 Z9 38 U1 0 U2 2 PU BAILLIERE TINDALL PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 1521-6934 J9 BEST PRACT RES CL OB JI Best Pract. Res. Clin. Obstet. Gynaecol. PD DEC PY 2000 VL 14 IS 6 BP 901 EP 918 DI 10.1053/beog.2000.0134 PG 18 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 393VF UT WOS:000166489800004 PM 11141340 ER PT J AU Dewaste, V Pouillon, V Moreau, C Shears, S Takazawa, K Erneux, C AF Dewaste, V Pouillon, V Moreau, C Shears, S Takazawa, K Erneux, C TI Cloning and expression of a cDNA encoding human inositol 1,4,5-trisphosphate 3-kinase C SO BIOCHEMICAL JOURNAL LA English DT Article DE calcium; phosphatidylinositol metabolism; signal transduction ID PROTEIN-KINASE-II; RAT-BRAIN; DIPHOSPHOINOSITOL PENTAKISPHOSPHATE; HEXAKISPHOSPHATE KINASE; MOLECULAR-CLONING; IDENTIFICATION; PURIFICATION; CALMODULIN; PHOSPHORYLATION; INS(1,4,5)P-3 AB Inositol 1,4,5-trisphosphate [Ins(1,4,5)P-3] 3-kinase catalyses the phosphorylation of Ins(1,4,5)P-3 to Ins(1,3,4,5)P-4. cDNAs encoding two isoenzymes of Ins(1,4,5)P-3 3-kinase (3-kinases A and B) have been described previously. In the present study, we report the cloning of a full-length 2052 bp cDNA encoding a third human isoenzyme of the Ins(1,4,5)P-3 3-kinase family, referred to as isoform C. This novel enzyme has a calculated molecular mass of 75.207 kDa and a K-m for Ins(1,4,5)P-3 of 6 muM. Northern-blot analysis showed the presence of a transcript of approx. 3.9 kb in various human tissues. Inositol trisphosphate 3-kinase C demonstrates enzymic activity when expressed in DH5 alphaF' bacteria or COS-7 cells. Calcium alone decreases the Ins(1,4,5)P-3 3-kinase activity of the 3-kinase C isoenzyme in transfected COS-7 cells. This inhibitory effect is reversed in the presence of calmodulin. The recombinant bacterial 3-kinase C can be adsorbed on calmodulin-Sepharose in the presence of calcium. The present data show that Ins(1,4,5)P-3 3-kinase C: (i) shares a conserved catalytic domain of about 275 amino acids with the two other mammalian isoforms, (ii) could be purified on a calmodulin-Sepharose column and (iii) could be distinguished from the A and B isoenzymes by the effects of calcium and of calmodulin. C1 Free Univ Brussels, IRIBHN, Interdisciplinary Res Inst, B-1070 Brussels, Belgium. NIEHS, Inositide Signaling Sect, Res Triangle Pk, NC 27709 USA. Tokyo Police Hosp, Tokyo 102, Japan. RP Erneux, C (reprint author), Free Univ Brussels, IRIBHN, Interdisciplinary Res Inst, Campus Erasme,Bldg C,808 Route Lennik, B-1070 Brussels, Belgium. NR 40 TC 47 Z9 50 U1 0 U2 0 PU PORTLAND PRESS PI LONDON PA 59 PORTLAND PLACE, LONDON W1N 3AJ, ENGLAND SN 0264-6021 J9 BIOCHEM J JI Biochem. J. PD DEC 1 PY 2000 VL 352 BP 343 EP 351 DI 10.1042/0264-6021:3520343 PN 2 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 385VC UT WOS:000166024400013 PM 11085927 ER PT J AU Dabholkar, M Thornton, K Vionnet, J Bostick-Bruton, F Yu, JJ Reed, E AF Dabholkar, M Thornton, K Vionnet, J Bostick-Bruton, F Yu, JJ Reed, E TI Increased mRNA levels of Xeroderma pigmentosum complementation group B (XPB) and Cockayne's syndrome complementation group B (CSB) without increased mRNA levels of multidrug-resistance gene (MDR1) or metallothionein-II (MT-II) in platinum resistant human ovarian cancer tissues SO BIOCHEMICAL PHARMACOLOGY LA English DT Article DE XPB; CSB; ovarian cancer; platinum compounds; MDR1; MT-II ID MISMATCH REPAIR DEFICIENCY; NUCLEOTIDE EXCISION-REPAIR; MESSENGER-RNA LEVELS; DNA-REPAIR; CELL-LINES; CISPLATIN SENSITIVITY; TRANSCRIPTION FACTOR; ACQUIRED-RESISTANCE; BTF2 TFIIH; GROUP-F AB Tumor tissue specimens from human ovarian cancer patients were assessed for relative mRNA abundance levels of several genes thought to be involved in the development of in vitro drug resistance in this disease. Higher mRNA levels of Xeroderma pigmentosum group B (XPB), which links DNA repair with DNA transcription, and of Cockayne's syndrome group B (CSB), which is essential for gene-specific repair, were observed in tumor tissues that were clinically resistant to platinum-based chemotherapy, as compared with tissues from patients responding to therapy. In a cohort of 27 patients, mRNA levels of XPB averaged 5-fold higher in platinum-resistant tumors (P = 0.001); and for CSB, mRNA levels averaged 6-fold higher but with greater variability (P = 0.033). Concurrently, these platinum-resistant tumor tissues did not exhibit significantly higher mRNA levels for the MDR1 (multidrug-resistance) gene (P = 0.134) or of the metallothionein-II (MT-II) gene (P = 0.598). Since these platinum-resistant tumors also show higher mRNA levels of ERCC1 and XPA, platinum resistance appears to be associated with concurrent up-regulation of four genes (XPA, ERCC1, XPB, and CSB). These four genes participate in DNA damage excision activity, gene-specific repair, and linkage of DNA repair with DNA transcription. These data suggest that concurrent up-regulation of genes involved in nucleotide excision repair may be important in clinical resistance to platinum-based chemotherapy in this disease. (C) 2000 Elsevier Science Inc. C1 NCI, Med Branch, Med Ovarian Canc Sect, NIH, Bethesda, MD 20892 USA. RP Reed, E (reprint author), NCI, Med Branch, Med Ovarian Canc Sect, NIH, Bldg 10,Rm 12N226, Bethesda, MD 20892 USA. NR 45 TC 28 Z9 31 U1 1 U2 4 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0006-2952 J9 BIOCHEM PHARMACOL JI Biochem. Pharmacol. PD DEC 1 PY 2000 VL 60 IS 11 BP 1611 EP 1619 DI 10.1016/S0006-2952(00)00448-2 PG 9 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 375QU UT WOS:000165413400008 PM 11077043 ER PT J AU Brambilla, E Munoz-Sanchez, JL Waalkes, MP Albores, A AF Brambilla, E Munoz-Sanchez, JL Waalkes, MP Albores, A TI Effect of surgically induced cholestasis on the levels of hepatic zinc and metallothionein in rat liver SO BIOLOGICAL TRACE ELEMENT RESEARCH LA English DT Article DE zinc; metallothionein; cholestasis ID EXCESS ZINC; PROGRESSIVE CHOLESTASIS; SERUM; HOMEOSTASIS; DISEASE; COPPER AB Early effects of experimental cholestasis on the homeostasis of zinc (Zn) and metallothionein (MT) were studied in rats which had undergone bile duct ligation for 0, 3, 6, 9, 12, 16, 20, and 24 h. Transient increases in hepatic Zn levels were observed at 9 h but returned to control values at 12 h. Serum Zn levels increased at 24 h. Cholestasis was confirmed by increased serum alkaline phosphatase (AP) activity. MT increased at 3 h and reached a maximum level at 12 h and remained elevated even at 24 h after the onset of experimental cholestasis. No hepatocellular damage was detected according to the results of alanine aminotransferase (ALT) activities in serum. These results shown that the increases in Zn observed in liver are related to bile stagnation rather than to a hepatocellular damage and that increased MT occurs concurrently with increased hepatic Zn. These observations suggest that the cellular levels of Zn in cholestasis is regulated by homeostatic mechanisms, of which one could be mediated by MT. C1 NIEHS, Inorgan Carcinogenesis Sect, Comparat Carcinogenesis Lab, NCI, Res Triangle Pk, NC 27706 USA. Benenerita Univ, Fac Ciencias Quim, Autonoma Puebla, Mexico. Inst Politecn Nacl, Escuela Nacl Ciencias Biol, Mexico City, DF, Mexico. IPN, Ctr Invest & Estud Avanzados, Dept Farmacol & Toxicol, Secc Toxicol Ambiental, Mexico City 07738, DF, Mexico. RP Brambilla, E (reprint author), NIEHS, Inorgan Carcinogenesis Sect, Comparat Carcinogenesis Lab, NCI, 111 Alexander Dr,Bldg 101,SC MD FO-09,RM F095, Res Triangle Pk, NC 27706 USA. NR 33 TC 3 Z9 3 U1 1 U2 4 PU HUMANA PRESS INC PI TOTOWA PA 999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USA SN 0163-4984 J9 BIOL TRACE ELEM RES JI Biol. Trace Elem. Res. PD WIN PY 2000 VL 78 IS 1-3 BP 255 EP 264 DI 10.1385/BTER:78:1-3:255 PG 10 WC Biochemistry & Molecular Biology; Endocrinology & Metabolism SC Biochemistry & Molecular Biology; Endocrinology & Metabolism GA 415UK UT WOS:000167741800022 PM 11314983 ER PT J AU Dunson, DB Dinse, GE AF Dunson, DB Dinse, GE TI Distinguishing effects on tumor multiplicity and growth rate in chemoprevention experiments SO BIOMETRICS LA English DT Article DE Bayesian analysis; breast cancer; Gibbs sampler; mammary tumors; mixture model; palpable tumors; tumorigenicity experiment ID STATISTICAL-ANALYSIS; CANCER; MODELS AB In some types of cancer chemoprevention experiments and short-term carcinogenicity bioassays, the data consist of the number of observed tumors per animal and the times at which these tumors were first detected. In such studies, there is interest in distinguishing between treatment effects on the number of tumors induced by a known carcinogen and treatment effects on the tumor growth rate. Since animals may die before all induced tumors reach a detectable size, separation of these effects can be difficult. This paper describes a flexible parametric model for data of this type. Under our model, the tumor detection times are realizations of a delayed Poisson process that is characterized by the age-specific tumor induction rate and a random latency interval between tumor induction and detection. The model accommodates distinct treatment and animal-specific effects on the number of induced tumors (multiplicity) and the time to tumor detection (growth rate). A Gibbs sampler is developed for estimation of the posterior distributions of the parameters. The methods are illustrated through application to data from a breast cancer chemoprevention experiment. C1 NIEHS, Biostat Branch, Res Triangle Pk, NC 27709 USA. RP Dunson, DB (reprint author), NIEHS, Biostat Branch, MD A3-03,POB 12233, Res Triangle Pk, NC 27709 USA. NR 19 TC 5 Z9 6 U1 2 U2 3 PU INTERNATIONAL BIOMETRIC SOC PI WASHINGTON PA 1441 I ST, NW, SUITE 700, WASHINGTON, DC 20005-2210 USA SN 0006-341X J9 BIOMETRICS JI Biometrics PD DEC PY 2000 VL 56 IS 4 BP 1068 EP 1075 DI 10.1111/j.0006-341X.2000.01068.x PG 8 WC Biology; Mathematical & Computational Biology; Statistics & Probability SC Life Sciences & Biomedicine - Other Topics; Mathematical & Computational Biology; Mathematics GA 383FN UT WOS:000165872600013 PM 11129462 ER PT J AU Baker, SG AF Baker, SG TI Identifying combinations of cancer markers for further study as triggers of early intervention SO BIOMETRICS LA English DT Article DE biomarker; bootstrap; diagnostic tests; discrimination; early detection; logistic regression; PSA; ROC curve; screening; sensitivity; specificity; validation ID PROSTATE-CANCER; BREAST-CANCER; PREDICTION AB In many long-term clinical trials or cohort studies, investigators repeatedly collect and store tissue or serum specimens and later test specimens from cancer cases and a random sample of controls for potential markers for cancer. An important question is what combination, if any, of the molecular markers should be studied in a future trial as a trigger for early intervention. To answer this question, we summarized the performance of various combinations using Receiver Operating Characteristic (ROC) curves, which plot true versus false positive rates. To construct the ROC curves, we proposed a new class of nonparametric algorithms which extends the ROC paradigm to multiple tests. We fit various combinations of markers to a training sample and evaluated the performance in a test sample using a target region based on a utility function. We applied the methodology to the following markers for prostate cancer, the last value of total prostate-specific antigen (PSA), the last ratio of total to free PSA, the last slope of total PSA, and the last slope of the ratio. In the test sample, the ROC curve for last total PSA was slightly closer to the target region than the ROC curve for a combination of four markers. in a separate validation sample, the ROC curve for last total PSA intersected the target region in 77% of bootstrap replications, indicating some promise for further study. We also discussed sample size calculations. C1 NCI, Biometry Res Grp, Div Canc Prevent, Bethesda, MD 20892 USA. RP Baker, SG (reprint author), NCI, Biometry Res Grp, Div Canc Prevent, EPN 344,6130 Execut Blvd MSC 7354, Bethesda, MD 20892 USA. FU NCI NIH HHS [CA70227]; NHLBI NIH HHS [HL57288]; NIGMS NIH HHS [GM54438] NR 21 TC 64 Z9 65 U1 0 U2 2 PU INTERNATIONAL BIOMETRIC SOC PI WASHINGTON PA 1441 I ST, NW, SUITE 700, WASHINGTON, DC 20005-2210 USA SN 0006-341X J9 BIOMETRICS JI Biometrics PD DEC PY 2000 VL 56 IS 4 BP 1082 EP 1087 DI 10.1111/j.0006-341X.2000.01082.x PG 6 WC Biology; Mathematical & Computational Biology; Statistics & Probability SC Life Sciences & Biomedicine - Other Topics; Mathematical & Computational Biology; Mathematics GA 383FN UT WOS:000165872600015 PM 11129464 ER PT J AU Hauptmann, M Wellmann, J Lubin, JH Rosenberg, PS Kreienbrock, L AF Hauptmann, M Wellmann, J Lubin, JH Rosenberg, PS Kreienbrock, L TI Analysis of exposure-time-response relationships using a spline weight function SO BIOMETRICS LA English DT Article DE bootstrap; case-control study; exposure history; exposure-time-response analysis; latency; lung cancer; smoking; splines; time since exposure; timing of exposure ID CIGARETTE-SMOKING AB To examine the time-dependent effects of exposure histories on disease, we estimate a weight function within a generalized linear model. The shape of the weight function, which is modeled as a cubic B-spline, gives information about the impact of exposure increments at different times on disease risk. The method is evaluated in a simulation study and is applied to data on smoking histories and lung cancer from a recent case-control study in Germany. C1 Univ Munster, Inst Epidemiol & Social Med, D-48129 Munster, Germany. Hannover Vet Sch, Inst Epidemiol & Biometry, D-30559 Hannover, Germany. GSF, Natl Res Ctr Environm & Hlth, Inst Epidemiol, D-85764 Neuherberg, Germany. NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. RP Hauptmann, M (reprint author), NCI, Div Canc Epidemiol & Genet, 6120 Execut Blvd,EPS 7089, Bethesda, MD 20892 USA. NR 10 TC 25 Z9 25 U1 1 U2 5 PU INTERNATIONAL BIOMETRIC SOC PI WASHINGTON PA 1441 I ST, NW, SUITE 700, WASHINGTON, DC 20005-2210 USA SN 0006-341X J9 BIOMETRICS JI Biometrics PD DEC PY 2000 VL 56 IS 4 BP 1105 EP 1108 DI 10.1111/j.0006-341X.2000.01105.x PG 4 WC Biology; Mathematical & Computational Biology; Statistics & Probability SC Life Sciences & Biomedicine - Other Topics; Mathematical & Computational Biology; Mathematics GA 383FN UT WOS:000165872600018 PM 11129467 ER PT J AU Proschan, MA Wittes, J AF Proschan, MA Wittes, J TI An improved double sampling procedure based on the variance SO BIOMETRICS LA English DT Article DE adaptive methods; double sampling; internal pilot studies; Stein's procedure AB Sample size calculations for a continuous outcome require specification of the anticipated variance; inaccurate specification can result in an underpowered or overpowered study. For this reason, adaptive methods whereby sample size is recalculated using the variance of a subsample have become increasingly popular. The first proposal of this type (Stein, 1945, Annals of Mathematical Statistics 16, 243-258) used all of the data to estimate the mean difference but only the first stage data to estimate the variance. Stein's procedure is not commonly used because many people perceive it as ignoring relevant data. This is especially problematic when the first stage sample size is small, as would be the case if the anticipated total sample size were small. A more naive approach uses in the denominator of the final test statistic the variance estimate based on all of the data. Applying the Helmert transformation, we show why this naive approach underestimates the true variance and how to construct an unbiased estimate that uses all of the data. We prove that the type I error rate of our procedure cannot exceed alpha. C1 NHLBI, Off Biostat Res, Rockledge Ctr 2, Bethesda, MD 20892 USA. Stat Collaborat, Washington, DC 20036 USA. RP Proschan, MA (reprint author), NHLBI, Off Biostat Res, Rockledge Ctr 2, Bldg 10,6701 Rockledge Dr,MSC 7938, Bethesda, MD 20892 USA. FU NCRR NIH HHS [2R44-RR08524-02] NR 7 TC 24 Z9 24 U1 0 U2 0 PU INTERNATIONAL BIOMETRIC SOC PI WASHINGTON PA 1441 I ST, NW, SUITE 700, WASHINGTON, DC 20005-2210 USA SN 0006-341X J9 BIOMETRICS JI Biometrics PD DEC PY 2000 VL 56 IS 4 BP 1183 EP 1187 DI 10.1111/j.0006-341X.2000.01183.x PG 5 WC Biology; Mathematical & Computational Biology; Statistics & Probability SC Life Sciences & Biomedicine - Other Topics; Mathematical & Computational Biology; Mathematics GA 383FN UT WOS:000165872600029 PM 11129477 ER PT J AU Yu, H Rothman, RB Dersch, CM Partilla, JS Rice, KC AF Yu, H Rothman, RB Dersch, CM Partilla, JS Rice, KC TI Uptake and release effects of diethylpropion and its metabolites with biogenic amine transporters SO BIOORGANIC & MEDICINAL CHEMISTRY LA English DT Article ID COCAINE AB Three metabolites of diethylpropion (1), (+/-)-2-ethylamino-1-phenyl-propan-1-one (2), (1 R,2S)-(-)-N,N-diethylnorephedrine (3a) and (1S,2R)-(-)-N,N-diethylnorephedrine (3b) were synthesized. Their uptake and release effects with biogenic amine transporters were evaluated. A major finding of this study is that the in vivo activity of diethylpropion on biogenic amine transporters is most likely due to metabolite 2 as diethylpropion (1) and the metabolites 3a and 3b showed little or no effect in the assays studied. These studies also revealed that 2 acted as a substrate at the norepinephrine (IC50=99nM) and serotonin transporters (IC50 = 2118 nM) and an uptake inhibitor at the dopamine transporter (IC50 = 1014 nM). The potent action of 2 at the NE transporter supports the hypothesis that amphetamine-type subjective effects may be mediated in part by brain norepinephrine. (C) 2000 Published by Elsevier Science Ltd. C1 NIDDKD, Med Chem Lab, NIH, Bethesda, MD 20892 USA. NIDA, Intramural Res Program, Clin Psychopharmacol Sect, Baltimore, MD 21224 USA. RP Rice, KC (reprint author), NIDDKD, Med Chem Lab, NIH, Bldg 8,Room B1-22, Bethesda, MD 20892 USA. NR 11 TC 23 Z9 24 U1 0 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0968-0896 J9 BIOORGAN MED CHEM JI Bioorg. Med. Chem. PD DEC PY 2000 VL 8 IS 12 BP 2689 EP 2692 DI 10.1016/S0968-0896(00)00210-8 PG 4 WC Biochemistry & Molecular Biology; Chemistry, Medicinal; Chemistry, Organic SC Biochemistry & Molecular Biology; Pharmacology & Pharmacy; Chemistry GA 381JR UT WOS:000165759700003 PM 11131159 ER PT J AU Bertram, R Previte, J Sherman, A Kinard, TA Satin, LS AF Bertram, R Previte, J Sherman, A Kinard, TA Satin, LS TI The phantom burster model for pancreatic beta-cells SO BIOPHYSICAL JOURNAL LA English DT Article ID INSULIN-SECRETING CELLS; CYTOPLASMIC FREE CA2+; 2 CALCIUM CURRENTS; ELECTRICAL-ACTIVITY; (I) OSCILLATIONS; ACTION-POTENTIALS; CULTURE DURATION; EXCITABLE CELL; ATP/ADP RATIO; DYNAMIC-CLAMP AB Pancreatic beta -cells exhibit bursting oscillations with a wide range of periods. Whereas periods in isolated cells are generally either a few seconds or a few minutes, in intact islets of Langerhans they are intermediate (10-60 s). We develop a mathematical model for beta -cell electrical activity capable of generating this wide range of bursting oscillations. Unlike previous models, bursting is driven by the interaction of two slow processes, one with a relatively small time constant (16 s) and the other with a much larger time constant (1-2 min). Bursting on the intermediate time scale is generated without need for a slow process having an intermediate time constant, hence phantom bursting. The model suggests that isolated cells exhibiting a fast pattern may nonetheless possess slower processes that can be brought out by injecting suitable exogenous currents. Guided by this, we devise an experimental protocol using the dynamic clamp technique that reliably elicits islet-like, medium period oscillations from isolated cells. Finally, we show that strong electrical coupling between a fast burster and a slow burster can produce synchronized medium bursting, suggesting that islets may be composed of cells that are intrinsically either fast or slow, with few or none that are intrinsically medium. C1 Florida State Univ, Inst Mol Biophys, Tallahassee, FL 32306 USA. Penn State Univ, Sch Sci, Erie, PA 16563 USA. NIDDKD, Math Res Branch, NIH, Bethesda, MD 20892 USA. Virginia Commonwealth Univ, Dept Pharmacol & Toxicol, Richmond, VA 23298 USA. Virginia Commonwealth Univ, Dept Physiol, Richmond, VA 23298 USA. RP Bertram, R (reprint author), Florida State Univ, Inst Mol Biophys, Tallahassee, FL 32306 USA. FU NIDDK NIH HHS [R01DK46409] NR 60 TC 69 Z9 70 U1 0 U2 0 PU BIOPHYSICAL SOCIETY PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0006-3495 J9 BIOPHYS J JI Biophys. J. PD DEC PY 2000 VL 79 IS 6 BP 2880 EP 2892 PG 13 WC Biophysics SC Biophysics GA 381EH UT WOS:000165749000009 PM 11106596 ER PT J AU Gorokhov, A Perera, L Darden, TA Negishi, M Pedersen, LC Pedersen, LG AF Gorokhov, A Perera, L Darden, TA Negishi, M Pedersen, LC Pedersen, LG TI Heparan sulfate biosynthesis: A theoretical study of the initial sulfation step by N-deacetylase/N-sulfotransferase SO BIOPHYSICAL JOURNAL LA English DT Article ID PARTICLE MESH EWALD; CRYSTAL-STRUCTURE; ESTROGEN SULFOTRANSFERASE; IN-VITRO; COMPLEX; MECHANISMS; PROTEINS; WINGLESS; CELLS AB Heparan sulfate N-deacetylase/N-sulfotransferase (NDST) catalyzes the deacetylation and sulfation of N-acetyl-D-glucosamine residues of heparan sulfate, a key step in its biosynthesis. Recent crystallographic and mutational studies have identified several potentially catalytic residues of the sulfotransferase domain of this enzyme (Kakuta et al., 1999, J. Biol. Chem. 274:10673-10676). We have used the x-ray crystal structure of heparan sulfate N-sulfotransferase with 3'-phosphoadenosine 5'-phosphate to build a solution model with cofactor 3'-phosphoadenosine 5'-phosphosulfate (PAPS) and a model heparan sulfate ligand bound, and subsequently performed a 2-ns dynamics solution simulation. The simulation results confirm the importance of residues Glu(642), Lys(614), and Lys(833), with the possible involvement of Thr(617) and Thr(618), in binding PAPS. Additionally, Lys(676) is found in close proximity to the reaction site in our solvated structure. This study illustrates for the first time the possible involvement of water in the catalysis. Three water molecules were found in the binding site, where they are coordinated to PAPS, heparan sulfate, and the catalytic residues. C1 Univ N Carolina, Dept Chem, Chapel Hill, NC 27599 USA. NIEHS, Res Triangle Pk, NC 27709 USA. RP Pedersen, LG (reprint author), Univ N Carolina, Dept Chem, Chapel Hill, NC 27599 USA. RI perera, Lalith/B-6879-2012; Pedersen, Lee/E-3405-2013 OI perera, Lalith/0000-0003-0823-1631; Pedersen, Lee/0000-0003-1262-9861 FU NHLBI NIH HHS [HL03650] NR 33 TC 13 Z9 14 U1 1 U2 6 PU BIOPHYSICAL SOCIETY PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0006-3495 J9 BIOPHYS J JI Biophys. J. PD DEC PY 2000 VL 79 IS 6 BP 2909 EP 2917 PG 9 WC Biophysics SC Biophysics GA 381EH UT WOS:000165749000012 PM 11106599 ER PT J AU Perera, L Foley, C Darden, TA Stafford, D Mather, T Esmon, CT Pedersen, LG AF Perera, L Foley, C Darden, TA Stafford, D Mather, T Esmon, CT Pedersen, LG TI Modeling zymogen protein C SO BIOPHYSICAL JOURNAL LA English DT Article ID GAMMA-CARBOXYGLUTAMIC ACID; THROMBIN-THROMBOMODULIN COMPLEX; K-DEPENDENT PROTEINS; MESH EWALD METHOD; BETA-HYDROXYASPARTIC ACID; SITE-DIRECTED MUTAGENESIS; COAGULATION-FACTOR VIIA; FACTOR HOMOLOGY DOMAIN; HELICAL STACK DOMAINS; SOLUBLE TISSUE FACTOR AB A solution structure for the complete zymogen form of human coagulation protein C is modeled. The initial core structure is based on the x-ray crystallographic structure of the gamma -carboxyglutamic acid (Gla)-domainless activated form. The Gla domain (residues 1-48) is modeled from the x-ray crystal coordinates of the factor VIIa/tissue factor complex and oriented with the epidermal growth factor-1 domain to yield an initial orientation consistent with the x-ray crystal structure of porcine factor IXa. The missing C-terminal residues in the light chain (residues 147-157) and the activation peptide residues 158-169 were introduced using homology modeling so that the activation peptide residues directly interact with the residues in the calcium binding loop. Molecular dynamics simulations (Amber-particle-mesh-Ewald) are used to obtain the complete calcium-complexed solution structure. The individual domain structures of protein C in solution are largely unaffected by solvation, whereas the Gla-epidermal growth factor-1 orientation evolves to a form different from both factors VIIa and IXa. The solution structure of the zymogen protein C is compared with the crystal structures of the existing zymogen serine proteases: chymotrypsinogen, proproteinase, and prethrombin-2. Calculated electrostatic potential surfaces support the involvement of the serine protease calcium ion binding loop in providing a suitable electrostatic environment around the scissile bond for IIa/thrombomodulin interaction. C1 Univ N Carolina, Dept Chem, Chapel Hill, NC 27599 USA. NIEHS, Res Triangle Pk, NC 27709 USA. Univ N Carolina, Dept Biol, Chapel Hill, NC 27599 USA. Oklahoma Med Res Fdn, Howard Hughes Med Inst, Oklahoma City, OK 73104 USA. RP Pedersen, LG (reprint author), Univ N Carolina, Dept Chem, Chapel Hill, NC 27599 USA. RI perera, Lalith/B-6879-2012; Pedersen, Lee/E-3405-2013; OI perera, Lalith/0000-0003-0823-1631; Pedersen, Lee/0000-0003-1262-9861; Foley, Charles/0000-0001-6578-9629 FU NHLBI NIH HHS [HL-06350] NR 86 TC 24 Z9 24 U1 0 U2 3 PU BIOPHYSICAL SOCIETY PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0006-3495 J9 BIOPHYS J JI Biophys. J. PD DEC PY 2000 VL 79 IS 6 BP 2925 EP 2943 PG 19 WC Biophysics SC Biophysics GA 381EH UT WOS:000165749000014 PM 11106601 ER PT J AU Stern, MD AF Stern, MD TI The model of Snyder et al. Does not simulate graded Ca2+ release from the cardiac sarcoplasmic reticulum in intact cells SO BIOPHYSICAL JOURNAL LA English DT Letter ID LOCAL-CONTROL; MUSCLE C1 NIA, Cardiovasc Sci Lab, NIH, Baltimore, MD 21224 USA. RP Stern, MD (reprint author), NIA, Cardiovasc Sci Lab, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 6 TC 3 Z9 3 U1 0 U2 1 PU BIOPHYSICAL SOCIETY PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0006-3495 J9 BIOPHYS J JI Biophys. J. PD DEC PY 2000 VL 79 IS 6 BP 3353 EP 3354 PG 2 WC Biophysics SC Biophysics GA 381EH UT WOS:000165749000051 PM 11203466 ER PT J AU Prieto, DA Harvey, LK Nelson, EL AF Prieto, DA Harvey, LK Nelson, EL TI Separation and purification of plasmid mixtures by continuous elution electrophoresis SO BIOTECHNIQUES LA English DT Article ID PREPARATIVE-SCALE PURIFICATION; DNA RESTRICTION FRAGMENTS; GEL-ELECTROPHORESIS; CHROMATOGRAPHY C1 NCI, Frederick, MD 21701 USA. RP Nelson, EL (reprint author), Univ Calif Irvine Med Surg 2, Rm 375B, Irvine, CA 92697 USA. FU PHS HHS [N01-C0-56000] NR 13 TC 3 Z9 3 U1 0 U2 2 PU EATON PUBLISHING CO PI NATICK PA 154 E. CENTRAL ST, NATICK, MA 01760 USA SN 0736-6205 J9 BIOTECHNIQUES JI Biotechniques PD DEC PY 2000 VL 29 IS 6 BP 1204 EP 1206 PG 3 WC Biochemical Research Methods; Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 382QG UT WOS:000165833300010 PM 11126121 ER PT J AU Post, RM Frye, MA Denicoff, KD Leverich, GS Dunn, RT Osuch, EA Speer, AM Obrocea, G Jajodia, K AF Post, RM Frye, MA Denicoff, KD Leverich, GS Dunn, RT Osuch, EA Speer, AM Obrocea, G Jajodia, K TI Emerging trends in the treatment of rapid cycling bipolar disorder: a selected review SO BIPOLAR DISORDERS LA English DT Review DE anticonvulsants; calcium channel blockers; carbamazepine; gabapentin; lamotrigine; lithium; nimodipine; repeated transcranial; magnetic stimulation (rTMS); tolerance; valproate ID TRANSCRANIAL MAGNETIC STIMULATION; LITHIUM TREATMENT; DOUBLE-BLIND; AFFECTIVE-ILLNESS; CONTROLLED TRIAL; MOOD DISORDERS; CALCIUM INFLUX; CARBAMAZEPINE; DEPRESSION; MANIA AB Recent evidence suggests that lithium therapy (even as supplemented by antidepressants and neuroleptics) is inadequate for the majority of patients with bipolar illness, and particularly those with rapid cycling. Valproate and carbamazepine have emerged as adjuncts and alternatives, but they, too, often require additional approaches with lithium, thyroid hormones, and other putative mood stabilizers, including nimodipine (and related dihydropyridine calcium channel blockers), lamotrigine, gabapentin? topiramate, and the atypical neuroleptics. Evaluating how these agents and the unimodal antidepressants are optimally applied and sequenced ill the treatment of bipolar illness with its multiple subtypes, patterns and comorbidities will require much future investigation and the development of new methodological clinical trial approaches. C1 NIMH, Biol Psychiat Branch, NIH, Bethesda, MD 20892 USA. RP Post, RM (reprint author), NIMH, Biol Psychiat Branch, NIH, Bldg 10,Room 3N212,10 Ctr Dr MSC 1272, Bethesda, MD 20892 USA. RI Osuch, Elizabeth/B-5009-2015 OI Osuch, Elizabeth/0000-0001-5946-1862 NR 67 TC 29 Z9 29 U1 0 U2 2 PU MUNKSGAARD INT PUBL LTD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 1398-5647 J9 BIPOLAR DISORD JI Bipolar Disord. PD DEC PY 2000 VL 2 IS 4 BP 305 EP 315 DI 10.1034/j.1399-5618.2000.020403.x PG 11 WC Clinical Neurology; Neurosciences; Psychiatry SC Neurosciences & Neurology; Psychiatry GA 428PQ UT WOS:000168470400003 PM 11252642 ER PT J AU Cheson, BD Bennett, JM Kantarjian, H Pinto, A Schiffer, CA Nimer, SD Lowenberg, B Beran, M de Witte, TM Stone, RM Mittelman, M Sanz, GF Wijermans, PW Gore, S Greenberg, PL AF Cheson, BD Bennett, JM Kantarjian, H Pinto, A Schiffer, CA Nimer, SD Lowenberg, B Beran, M de Witte, TM Stone, RM Mittelman, M Sanz, GF Wijermans, PW Gore, S Greenberg, PL TI Report of an international working group to standardize response criteria for myelodysplastic syndromes SO BLOOD LA English DT Editorial Material ID CHRONIC MYELOGENOUS LEUKEMIA; INTERFERON-ALPHA THERAPY; MARROW TRANSPLANTATION; CLASSIFICATION; EVOLUTION; SURVIVAL; DISEASE AB Standardized criteria for assessing response are essential to ensure comparability among clinical trials for patients with myelodysplastic syndromes (MDS). An international working group of experienced clinicians involved in the management of patients with MDS reviewed currently used response definitions and developed a uniform set of guidelines for future clinical trials in MDS. The MDS differ from many other hematologic malignancies in their chronicity and the morbidity and mortality caused by chronic cytopenias, often without disease progression to acute myeloid leukemia. Whereas response rates may be an important endpoint for phase 2 studies of new agents and may assist regulatory agencies in their evaluation and approval processes, an important goal of clinical trials in MDS should be to prolong patient survival. Therefore, these response criteria reflected 2 sets of goals in MDS: altering the natural history of the disease and alleviating disease-related complications with improved quality of life. It is anticipated that the recommendations presented will require modification as more is learned about the molecular biology and genetics of these disorders. Until then, it is hoped these guidelines will serve to improve communication among investigators and to ensure comparability among clinical trials. (C) 2000 by The American Society of Hematology. C1 NCI, Bethesda, MD 20892 USA. Univ Rochester, Ctr Canc, Rochester, NY 14627 USA. Univ Texas, MD Anderson Canc Ctr, Houston, TX 77030 USA. Ctr Riferimento Oncol, I-33081 Aviano, Italy. Wayne State Univ, Karmanos Canc Ctr, Detroit, MI USA. Mem Sloan Kettering Canc Ctr, New York, NY 10021 USA. Univ Rotterdam Hosp, Rotterdam, Netherlands. Univ Nijmegen Hosp, Rijmegen, Netherlands. Dana Farber Canc Inst, Boston, MA 02115 USA. Hasharon Hosp, Rabin Med Ctr, IL-49372 Petah Tiqwa, Israel. Tel Aviv Univ, IL-69978 Tel Aviv, Israel. Univ Valencia, Hosp La Fe, Valencia, Spain. Ziekenhuis The Hague, The Hague, Netherlands. Johns Hopkins Oncol Ctr, Baltimore, MD USA. Stanford Univ, Med Ctr, Palo Alto, CA 94304 USA. Vet Adm Hosp, Palo Alto, CA USA. RP Cheson, BD (reprint author), NCI, Execut Plaza N,Room 741, Bethesda, MD 20892 USA. RI Witte, T.J.M./L-4762-2015; OI SANZ, GUILLERMO/0000-0002-2767-8191 NR 22 TC 457 Z9 490 U1 1 U2 5 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1900 M STREET. NW SUITE 200, WASHINGTON, DC 20036 USA SN 0006-4971 J9 BLOOD JI Blood PD DEC 1 PY 2000 VL 96 IS 12 BP 3671 EP 3674 PG 4 WC Hematology SC Hematology GA 377LA UT WOS:000165514000002 PM 11090046 ER PT J AU Bronte, V Apolloni, E Cabrelle, A Ronca, R Serafini, P Zamboni, P Restifo, NP Zanovello, P AF Bronte, V Apolloni, E Cabrelle, A Ronca, R Serafini, P Zamboni, P Restifo, NP Zanovello, P TI Identification of a CD11b(+)/Gr-1(+)/CD31(+) myeloid progenitor capable of activating or suppressing CD8(+) T cells SO BLOOD LA English DT Article ID COLONY-STIMULATING FACTOR; MOUSE BONE-MARROW; ANTIGEN-PRESENTING CELLS; LIGAND-TREATED MICE; DENDRITIC CELLS; MACROPHAGE ACTIVATION; MURINE MACROPHAGES; NITRIC-OXIDE; IN-VITRO; GM-CSF AB Apoptotic death of CD8(+) T cells can be induced by a population of inhibitory myeloid cells that are double positive for the CD11b and Gr-1 markers. These cells are responsible for the immunosuppression observed in pathologies as dissimilar as tumor growth and overwhelming infections, or after immunization with viruses. The appearance of a CD11b(+)/Gr-1(+) population of inhibitory macrophages (iMacs) could be attributed to high levels of granulocyte-macrophage colony-stimulating factor (GM-CSF)in vivo. Deletion of iMacs in vitro or in vivo reversed the depression of CD8(+) T-cell function. We isolated iMacs from the spleens of immunocompromised mice and found that these cells were positive for CD31, ER-MP20 (Ly-6C), and ER-MP58, markers characteristic of granulocyte/monocyte precursors. Importantly, although iMacs retained their inhibitory properties when cultured in vitro in standard medium, suppressive functions could be modulated by cytokine exposure. Whereas culture with the cytokine interleukin 4 (IL-4) increased iMac inhibitory activity, these cells could be differentiated into a nonadherent population of fully mature and highly activated dendritic cells when cultured in the presence of IL-4 and GM CSF. A common CD31(+)/CD11b(+)/Gr-1(+) progenitor can th us give rise to cells capable of either activating or inhibiting the function of CD8(+) T lymphocytes, depending on the cytokine milieu that prevails during antigen-presenting cell maturation. (C) 2000 by The American Society of Hematology. C1 Univ Padua, Dept Oncol & Surg Sci, I-35128 Padua, Italy. NCI, Surg Branch, NIH, Bethesda, MD 20892 USA. RP Bronte, V (reprint author), Univ Padua, Dept Oncol & Surg Sci, Via Gattamelata 64, I-35128 Padua, Italy. RI Restifo, Nicholas/A-5713-2008; Serafini, Paolo/C-8195-2012; Bronte, Vincenzo/K-7902-2016; OI Serafini, Paolo/0000-0002-3651-9176; Bronte, Vincenzo/0000-0002-3741-5141; Restifo, Nicholas P./0000-0003-4229-4580 FU Intramural NIH HHS [Z01 BC010763-01, Z99 CA999999] NR 44 TC 336 Z9 354 U1 0 U2 15 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1900 M STREET. NW SUITE 200, WASHINGTON, DC 20036 USA SN 0006-4971 J9 BLOOD JI Blood PD DEC 1 PY 2000 VL 96 IS 12 BP 3838 EP 3846 PG 9 WC Hematology SC Hematology GA 377LA UT WOS:000165514000024 PM 11090068 ER PT J AU Yamashiro, S Wang, JM Yang, D Gong, WH Kamohara, H Yoshimura, T AF Yamashiro, S Wang, JM Yang, D Gong, WH Kamohara, H Yoshimura, T TI Expression of CCR6 and CD83 by cytokine-activated human neutrophils SO BLOOD LA English DT Article ID COLONY-STIMULATING FACTOR; DELAYED-TYPE HYPERSENSITIVITY; MONOCYTE CHEMOATTRACTANT PROTEIN-1; HUMAN POLYMORPHONUCLEAR NEUTROPHILS; PROGRAMMED CELL-DEATH; COMPLEX CLASS-II; RED-BLOOD-CELLS; MESSENGER-RNA; MONOCLONAL-ANTIBODY; DENDRITIC CELLS AB Polymorphonuclear leukocytes (PMNLs) are thought to be terminally differentiated, short-lived, and unable to actively synthesize new proteins or to interact with T cells, In the current study, it was found that PMNLs incubated with supernatants of phytohemagglutinin (PHA)stimulated peripheral blood mononuclear cells (PHA-sup) expressed high levels of CCR6 mRNA, Neutralization with IgG against several cytokines revealed that tumor necrosis factor (TNF)-alpha was largely responsible for the PHA-sup-induced CCR6 mRNA expression. Among recombinant cytokines, TNF-alpha induced high levels of CCR6 mRNA expression, whereas interferon (IFN)-gamma induced low levels. The 2 cytokines together exhibited a considerable synergy. Cytokine-activated PMNLs expressed functional CCR6, as detected by the binding of sodium iodide I 125-labeled liver and activation-regulated chemokine (LARC) and dose-dependent migration toward LARC, The induction of CCR6 suggested that these cytokine-activated PMNLs have more similarities with dendritic cells (DCs) that express CCR6 in an immature stage. In fact, the activation of PMNLs with TNF-alpha and IFN-gamma induced the expression of CD83, a dominant cell-surface marker of DCs, When PMNLs were activated with granulocyte macrophage-colony-stimulating factor, TNF-alpha, and IFN-gamma, these cells expressed CD40 and HLA-DR in addition to CD83. Taken together, PMNLs, under appropriate conditions, can undergo a differentiation process characterized by the acquisition of new phenotypes and functions, and such differentiated PMNLs may play more active roles in the adaptive immune response. (C) 2000 by The American Society of Hematology. C1 NCI, Frederick Canc Res & Dev Ctr, Mol Immunoregulat Lab, Frederick, MD 21702 USA. RP Yoshimura, T (reprint author), NCI, Frederick Canc Res & Dev Ctr, Mol Immunoregulat Lab, Bldg 559,Rm 1, Frederick, MD 21702 USA. NR 26 TC 100 Z9 102 U1 0 U2 1 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1900 M STREET. NW SUITE 200, WASHINGTON, DC 20036 USA SN 0006-4971 J9 BLOOD JI Blood PD DEC 1 PY 2000 VL 96 IS 12 BP 3958 EP 3963 PG 6 WC Hematology SC Hematology GA 377LA UT WOS:000165514000040 PM 11090084 ER PT J AU Salama, M Nevill, T Marcellus, D Parker, P Johnson, M Kirk, A Porter, D Giralt, S Levine, JE Drobyski, W Barrett, AJ Horowitz, M Collins, RH AF Salama, M Nevill, T Marcellus, D Parker, P Johnson, M Kirk, A Porter, D Giralt, S Levine, JE Drobyski, W Barrett, AJ Horowitz, M Collins, RH TI Donor leukocyte infusions for multiple myeloma SO BONE MARROW TRANSPLANTATION LA English DT Article DE multiple myeloma; donor leukocyte infusions; adoptive immunotherapy ID BONE-MARROW TRANSPLANTATION; GRAFT-VERSUS-MYELOMA; CHRONIC MYELOGENOUS LEUKEMIA; HOST-DISEASE; ADOPTIVE IMMUNOTHERAPY; LYMPHOCYTE INFUSION; RELAPSE; TRANSFUSIONS; CELLS AB Donor leukocyte infusion (DLI) has well-documented activity in CML, but the role of DLI in other diseases is less well defined. To evaluate the strategy in multiple myeloma (MM) we evaluated 25 MM patients from 15 centers who were treated with DLI, Patients with persistent or recurrent disease after allogeneic BMT received DLI from the original marrow donor (23 matched related, one mismatched family, and one matched unrelated). Chemotherapy was given before DLI in three patients, Two of 22 patients responded completely to DLI alone and three patients responded to the combination of DLI and chemotherapy. Nine patients who had not had sufficient disease control after DLI were given additional DLIs; five of these patients had either complete (two) or partial (three) responses. Thirteen of 25 evaluable patients developed acute GVHD and 11 of 21 evaluable patients developed chronic GVHD; all responders developed GVHD. No patients developed post-DLI pancytopenia. Four patients had responses which lasted >1 year after DLI, three patients had responses which lasted <1 year, and three patients had ongoing responses but with followup <1 year. In conclusion, DLI has anti-myeloma activity but the strategy is limited by no response or short duration of response in a significant percentage of patients and by significant GVHD in the majority of the responders. C1 Univ Texas, SW Med Ctr, Dallas, TX 75390 USA. Vancouver Hosp & Hlth Sci Ctr, Vancouver, BC V5Z 1M9, Canada. Johns Hopkins Univ Hosp, Baltimore, MD 21205 USA. City Hope, Duarte, CA USA. Baylor Sammons Canc Ctr, Dallas, TX USA. Univ Penn, Sch Med, Philadelphia, PA 19104 USA. Univ Texas, MD Anderson Canc Ctr, Houston, TX 77030 USA. Univ Michigan, Ann Arbor, MI 48109 USA. Med Coll Wisconsin, Milwaukee, WI 53226 USA. NHLBI, Bethesda, MD 20892 USA. Med Coll Wisconsin, Int Bone Marrow Transplant Registry, Milwaukee, WI 53226 USA. RP Collins, RH (reprint author), Univ Texas, SW Med Ctr, 5323 Harry Hines Blvd, Dallas, TX 75390 USA. NR 30 TC 118 Z9 123 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0268-3369 J9 BONE MARROW TRANSPL JI Bone Marrow Transplant. PD DEC PY 2000 VL 26 IS 11 BP 1179 EP 1184 DI 10.1038/sj.bmt.1702685 PG 6 WC Biophysics; Oncology; Hematology; Immunology; Transplantation SC Biophysics; Oncology; Hematology; Immunology; Transplantation GA 382WZ UT WOS:000165851500006 PM 11149728 ER PT J AU Lawrence, DA Harry, GJ AF Lawrence, DA Harry, GJ TI Environmental stressors and neuroimmunotoxicological processes - Introduction SO BRAIN BEHAVIOR AND IMMUNITY LA English DT Editorial Material ID CENTRAL-NERVOUS-SYSTEM; MESSENGER-RNA LEVELS; ALZHEIMERS-DISEASE; BRAIN; ACTIVATION; EXPRESSION; CELLS; MODEL; RATS; MICE C1 New York State Dept Hlth, Wadsworth Ctr Labs & Res, Lab Clin & Expt Endocrinol & Immunol, Albany, NY 12201 USA. NIEHS, Neurotoxicol Grp, Toxicol Lab, Res Triangle Pk, NC 27709 USA. RP Lawrence, DA (reprint author), New York State Dept Hlth, Wadsworth Ctr Labs & Res, Lab Clin & Expt Endocrinol & Immunol, Albany, NY 12201 USA. NR 31 TC 5 Z9 5 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0889-1591 J9 BRAIN BEHAV IMMUN JI Brain Behav. Immun. PD DEC PY 2000 VL 14 IS 4 BP 231 EP 238 DI 10.1006/brbi.2000.0607 PG 8 WC Immunology; Neurosciences SC Immunology; Neurosciences & Neurology GA 388HE UT WOS:000166174100001 PM 11120593 ER PT J AU Harry, GJ d'Hellencourt, CL Bruccoleri, A Schmechel, D AF Harry, GJ d'Hellencourt, CL Bruccoleri, A Schmechel, D TI Age-dependent cytokine responses: Trimethyltin hippocampal injury in wild-type, APOE knockout, and APOE4 mice SO BRAIN BEHAVIOR AND IMMUNITY LA English DT Article DE apolipoprotein E; GFAP; EB22; TNF alpha; MIP-1 alpha; TGF-beta 1 ID E-DEFICIENT MICE; NECROSIS-FACTOR-ALPHA; APOLIPOPROTEIN-E POLYMORPHISM; MOLECULE-1 GENE-EXPRESSION; ALZHEIMERS-DISEASE CERAD; DIFFERENT PLAQUE TYPES; CLOSED-HEAD INJURY; INTERFERON-GAMMA; MESSENGER-RNA; NERVOUS-SYSTEM AB In this study, the hippocampal neurotoxicant trimethyltin (TMT) was used to examine possible differential susceptibility associated with the apolipoprotein E genotype. Mice-wild type (C57BL6J), APOE knockout, and APOE4 transgenic-received either saline or TMT (2 mg/kg, ip) at either 21 days or 8 months of age. At both ages, similar mRNA levels were seen in the hippocampus across genotypes for ICAM-1, A20, and MAC-I. GFAP mRNA was higher in the APOE knockouts and APOE4 as compared to wild-type mice. Within 24 h, TMT produced cell death of hippocampal dentate granule neurons and mild astrogliosis in all animals. In 21-day-old mice, TMT exposure significantly increased mRNA levels for ICAM-1 and MIP-1 alpha in all genotypes. EB-22, GFAP, TNF alpha, and TGF-beta1 levels were significantly elevated in both wild-type and APOE knockout mice following TMT. At 8 months of age, genotype specific differences were observed. mRNA levels for GFAP, TNF beta, TNF alpha, and MIP-1 alpha were increased in both APOE knockout and APOE4 mice compared to wild-type mice. TMT exposure significantly increased mRNA levels for GFAP and MIP-1 alpha in all animals. TNF alpha mRNA levels were increased in wild-type and APOE4 mice while EB22 mRNA levels were increased in both the APOE knockout and APOE4 mice but not wild-type mice. These data suggest an age-dependent effect on both microglia early inflammatory responses to injury associated with the APOE genotype. (C) 2000 Academic Press. C1 NIEHS, Neurotoxicol Grp, Toxicol Lab, Res Triangle Pk, NC 27709 USA. Joseph & Kathleen Bryan Alzheimers Dis Res Ctr, Dept Med Neurol, Durham, NC 27710 USA. RP Harry, GJ (reprint author), NIEHS, Neurotoxicol Grp, Toxicol Lab, POB 12233, Res Triangle Pk, NC 27709 USA. FU NIA NIH HHS [5P50 AG-05128]; NIEHS NIH HHS [Y01-ES-40290] NR 80 TC 19 Z9 19 U1 0 U2 5 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0889-1591 J9 BRAIN BEHAV IMMUN JI Brain Behav. Immun. PD DEC PY 2000 VL 14 IS 4 BP 288 EP 304 DI 10.1006/brbi.2000.0606 PG 17 WC Immunology; Neurosciences SC Immunology; Neurosciences & Neurology GA 388HE UT WOS:000166174100005 PM 11120597 ER PT J AU Mandavilli, BS Ali, SF Van Houten, B AF Mandavilli, BS Ali, SF Van Houten, B TI DNA damage in brain mitochondria caused by aging and MPTP treatment SO BRAIN RESEARCH LA English DT Article DE aging; dopamine; MPTP; oxidative stress; DNA damage; mitochondria ID PARKINSONS-DISEASE; OXIDATIVE DAMAGE; SUBSTANTIA-NIGRA; HYDROGEN-PEROXIDE; MOUSE-BRAIN; 1-METHYL-4-PHENYL-1,2,3,6-TETRAHYDROPYRIDINE MPTP; STRIATAL DOPAMINE; NUCLEAR-DNA; COMPLEX-I; ATP LOSS AB 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyrine (MPTP) treatment leads to marked depletion of dopamine (DA) levels in the nigrostriatal pathway and dopaminergic neuronal degeneration in caudate-putamen and substantia nigra. MPTP is believed to inhibit complex I of the electron transport system leading to the generation of reactive oxygen species. We sought to test the hypotheses that MPTP treatment: (1) leads to dopamine depletion; (2) causes extensive mitochondrial DNA damage, and (3) that these effects would be age dependent. The levels of dopamine and its metabolites, DOPAC and HVA were analyzed by HPLC equipped with electrochemical detection. DNA damage was measured by quantitative PCR in both mitochondrial and nuclear (beta -polymerase) targets from the caudate-putamen, substantia nigra and cerebellum regions of control and MPTP-treated mice. The age groups studied were 22 days and 12 months. MPTP produced no significant effect on the levels of dopamine and its metabolites in young mice whereas in old, there was a significant decrease in this neurotransmitter system after MPTP administration. These 12-month-old mice, when compared to the young mice, showed a significant increase in mitochondrial DNA damage in the caudate-putamen and cerebellum. The latter region also displayed a significant increase in DNA damage in a nuclear gene. After treatment with MPTP, there was an age-dependent increase in DNA damage in mitochondria of the caudate-putamen while there was no significant DNA damage in the nuclear target. MPTP treatment led to damage in both mitochondrial and nuclear DNA of the substantia nigra, while there was no damage in either mitochondria or nucleus in cerebellum which was used as a negative control. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIEHS, Mol Genet Lab, NIH, Res Triangle Pk, NC 27709 USA. Natl Ctr Toxicol Res, Div Neurotoxicol, Neurochem Lab, Jefferson, AR 72079 USA. RP Van Houten, B (reprint author), NIEHS, Mol Genet Lab, NIH, 111 Alexander Dr,Box 12233, Res Triangle Pk, NC 27709 USA. FU NIA NIH HHS [2PO1AG10514] NR 63 TC 51 Z9 51 U1 0 U2 3 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0006-8993 J9 BRAIN RES JI Brain Res. PD DEC 1 PY 2000 VL 885 IS 1 BP 45 EP 52 DI 10.1016/S0006-8993(00)02926-7 PG 8 WC Neurosciences SC Neurosciences & Neurology GA 380TE UT WOS:000165718900006 PM 11121528 ER PT J AU Partilla, JS Dersch, CM Yu, H Rice, KC Rothman, RB AF Partilla, JS Dersch, CM Yu, H Rice, KC Rothman, RB TI Neurochemical neutralization of amphetamine-type stimulants in rat brain by the indatraline analog (-)-HY038 SO BRAIN RESEARCH BULLETIN LA English DT Article DE amphetamine; neurotransmitter; MDMA; methampletamine; indatraline ID BIOGENIC-AMINE TRANSPORTERS; DOPAMINE; NOREPINEPHRINE; SEROTONIN; BINDING; INHIBITORS; GBR12909; REUPTAKE; MONKEYS AB Amphetamine-type stimulants are substrates for the proteins that serve as transporters for the biogenic amines dopamine (DA), serotonin (5HT), and norepinephrine (NE) and release these neurotransmitters from neurons located in the peripheral and central nervous system. Using indatraline as a lead compound, we sought to develop a long-acting depot medication that would neutralize the deleterious effects of amphetamine-type stimulants. our first efforts produced (+/-)-HY038, and its two stereoisomers, which are hydroxy-substituted analog of indatraline. The K-i values for [H-3]DA reuptake inhibition by (-)-HY038 and (+)-HY038 were 3.2 +/- 0.1 and 32 +/- 1 nM. Similar results were obtained for [H-3]5HT reuptake inhibition. (-)-HY038 and (+)-HY038 were slightly less potent at inhibiting [H-3]NE reuptake (K-i values of 20 +/- 2 and 159 +/- 12 nM), Low doses of (-)-HY038 blunted the ability of AMPH to release [H-3]DA by shifting the AMPH dose-response curve to the right in a dose-dependent manner. (-)-HY038 also inhibited the ability of (+)-methamphetamine and (+/-)-3,4-methylenedioxymethamphetamine ((+/-)-MDMA) to release [H-3]DA. Low doses of (-)-HY038 blunted the ability of these stimulants to release [H-3]NE and [H-3]5HT by shifting their dose-response! curves to the right in a manner similar to that seen for inhibition of [H-3]DA release, These data indicate that (-)-HY038 inhibits the ability of AMPH, (+)-methamphetamine and (+/-)-MDMA to release DA, NE, and 5HT and therefore might have the potential to neutralize the neurotoxic and cardiovascular side-effects of substrate-type stimulants. (C) 2001 Elsevier Science Inc. C1 NIDA, Clin Psychopharmacol Sect, Intramural Res Program, NIH, Baltimore, MD 21224 USA. NIDDKD, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. RP Rothman, RB (reprint author), NIDA, Clin Psychopharmacol Sect, Intramural Res Program, NIH, POB 5180,5500 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 18 TC 10 Z9 10 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0361-9230 J9 BRAIN RES BULL JI Brain Res. Bull. PD DEC PY 2000 VL 53 IS 6 BP 821 EP 826 DI 10.1016/S0361-9230(00)00419-6 PG 6 WC Neurosciences SC Neurosciences & Neurology GA 403UH UT WOS:000167060500013 PM 11179849 ER PT J AU Frost, DO Cadet, JL AF Frost, DO Cadet, JL TI Effects of methamphetamine-induced neurotoxicity on the development of neural circuitry: a hypothesis SO BRAIN RESEARCH REVIEWS LA English DT Review DE dopamine; serotonin; neuroplasticity; free radicals; apoptosis ID METHYL-D-ASPARTATE; NEURONS IN-VITRO; EXHIBIT DIFFERENTIAL VULNERABILITY; QUANTITATIVE MORPHOMETRIC ANALYSIS; OCULAR DOMINANCE PLASTICITY; ENHANCES NEURITE OUTGROWTH; RATS SOMATOSENSORY CORTEX; DISMUTASE TRANSGENIC MICE; HYPOXIC-ISCHEMIC INJURY; HUMAN SUBSTANTIA-NIGRA AB Exposure of the developing brain to methamphetamine has well-studied biochemical and behavioral consequences. We review: (1) the effects of methamphetamine on mature serotonergic and dopaminergic pathways; (2) the mechanisms of methamphetamine neurotoxicity and (3) the role of serotonergic and dopaminergic signaling in sculpting developing neural circuitry. Consideration of these data suggest the types of neural circuit alterations that may result from exposure of the developing brain to methamphetamine and that may underlie functional defects. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Univ Maryland, Sch Med, Dept Pharmacol & Expt Therapeut, Baltimore, MD 21201 USA. Univ Maryland, Sch Med, Neurosci Program, Baltimore, MD 21201 USA. NIDA, Mol Neuropsychiat Sect, Baltimore, MD USA. RP Frost, DO (reprint author), Univ Maryland, Sch Med, Dept Pharmacol & Expt Therapeut, 655 W Baltimore St, Baltimore, MD 21201 USA. FU NEI NIH HHS [R01-EY11434-01A1] NR 238 TC 54 Z9 54 U1 2 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0165-0173 J9 BRAIN RES REV JI Brain Res. Rev. PD DEC PY 2000 VL 34 IS 3 BP 103 EP 118 DI 10.1016/S0165-0173(00)00042-4 PG 16 WC Neurosciences SC Neurosciences & Neurology GA 383FK UT WOS:000165872300001 PM 11113502 ER PT J AU Rao, GN Ney, E Herbert, RA AF Rao, GN Ney, E Herbert, RA TI Effect of melatonin and linolenic acid on mammary cancer in transgenic mice with c-neu breast cancer oncogene SO BREAST CANCER RESEARCH AND TREATMENT LA English DT Article DE breast cancer; c-neu transgenic mice; melatonin; linolenic acid; flaxseed oil; IGF-1 concentrations ID GROWTH-FACTOR-I; RETINOID ANALOGS; DIET NTP-2000; BEARING; MODELS; TISSUE; RATS; CARCINOGENESIS; SUPPRESSION; GLUTATHIONE AB Breast cancer is one of the most common cancers and is a leading cause of mortality in women. The TG.NK transgenic mouse line expresses the c-neu breast cancer oncogene under the control of a MMTV promoter and appears to be a useful animal model for evaluation of intervention strategies to delay/prevent breast cancer. Fiber-rich nonpurified diet (NTP-2000) and some retinoid analogues have been shown to significantly delay the development of mammary cancer in the TG.NK model. Four-week-old hemizygous TG.NK female mice with MMTV/c-neu oncogene fed NTP-2000 diet were gavaged with 0.05-0.2 ml of flaxseed oil as the source of omega -3 rich PUFA, or melatonin at 50-200 mg/kg or a combination of 0.10 ml flaxseed oil and 50 mg/kg melatonin in a gavage volume of 0.2 ml per mouse with corn oil as the vehicle for 30 weeks. The time course of the mammary tumor incidence pattern was advanced by flaxseed oil compared to the control. At the high dose (0.2 ml) of flaxseed oil, when the omega -6: omega -3 PUFA ratio was closer to 1, there was some delay in the growth of mammary tumors. Melatonin delayed the appearance of palpable tumors and the growth of the tumors with a dose-related statistically significant negative trend for the incidence of tumors. The combination of flaxseed oil and melatonin caused a significant decrease in the number of tumors and tumor weight per mouse compared to the control and to flaxseed oil but not to melatonin alone. Flaxseed oil may delay the growth of mammary tumors if the omega -6:omega -3 PUFA ratio of fat consumed is closer to 1. Melatonin has the potential to markedly delay the appearance of palpable mammary tumors. Studies are in progress with the TG.NK mouse model to understand the histological and molecular changes associated with the dose-response pattern of mammary tumor incidence and growth after treatment with a broad range of doses of melatonin. C1 NIEHS, Environm Toxicol Program, NIH, Res Triangle Pk, NC 27709 USA. RP Rao, GN (reprint author), NIEHS, Environm Toxicol Program, NIH, MD B3-08,POB 12233, Res Triangle Pk, NC 27709 USA. EM rao@niehs.nih.gov NR 46 TC 27 Z9 29 U1 0 U2 4 PU SPRINGER PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 0167-6806 J9 BREAST CANCER RES TR JI Breast Cancer Res. Treat. PD DEC PY 2000 VL 64 IS 3 BP 287 EP 296 DI 10.1023/A:1026552405042 PG 10 WC Oncology SC Oncology GA 379FH UT WOS:000165630800007 PM 11200779 ER PT J AU Sausville, EA Johnson, JI AF Sausville, EA Johnson, JI TI Molecules for the millennium: how will they look? New drug discovery year 2000 SO BRITISH JOURNAL OF CANCER LA English DT Review DE drug discovery paradigm; molecular targets; anticancer therapeutics; in vitro screens ID TUMOR-CELL LINES; CANCER; SENSITIVITY; INHIBITORS; CHEMOTHERAPY; EXPRESSION; TARGETS; COMPLEX; GROWTH; SCREEN AB A new approach to cancer drug discovery targets molecules important in cancer pathogenesis. This approach is thought to be of greater promise than the antiproliferative screens which discovered cytotoxic agents and dominated cancer drug discovery for 60 years. However, one cannot lose sight of the fact that these targets exist in the cellular environment consisting of many additional influences on target function, and that effective drug treatment will take into account drug uptake, metabolism and elimination at the level of the cell as well as the organism. A key goal is to define for the new millennium a path to cancer drug discovery and development which accounts for the cancer cell phenotype in its totality rather than as arising solely from single molecular targets. The US National Cancer Institute maintains a cell-based drug discovery screen which can define a context for drug action in the milieu of more than 300 molecular targets and thousands of gene expression patterns which have been measured in the 60 human tumour cell lines which comprise the screening panel. The challenge of the millennium will be addressed by molecules active against defined targets but with selectivity of action occurring in the milieu of deregulated cancer cell biology in all its aspects. (C) 2000 Cancer Research Campaign http://www.bjcancer.com. C1 NCI, Div Canc Treatment & Diag, Dev Therapeut Program, Rockville, MD 20852 USA. RP Sausville, EA (reprint author), NCI, Div Canc Treatment & Diag, Dev Therapeut Program, 6130 Execut Blvd,Suite 8000, Rockville, MD 20852 USA. NR 25 TC 10 Z9 11 U1 1 U2 1 PU CHURCHILL LIVINGSTONE PI EDINBURGH PA JOURNAL PRODUCTION DEPT, ROBERT STEVENSON HOUSE, 1-3 BAXTERS PLACE, LEITH WALK, EDINBURGH EH1 3AF, MIDLOTHIAN, SCOTLAND SN 0007-0920 J9 BRIT J CANCER JI Br. J. Cancer PD DEC PY 2000 VL 83 IS 11 BP 1401 EP 1404 DI 10.1054/bjoc.2000.1473 PG 4 WC Oncology SC Oncology GA 378UL UT WOS:000165603000001 PM 11076644 ER PT J AU Bellahcene, A Van Riet, I de Greef, C Antoine, N Young, MF Van Camp, B Castronovo, V AF Bellahcene, A Van Riet, I de Greef, C Antoine, N Young, MF Van Camp, B Castronovo, V TI Bone sialoprotein mRNA and protein expression in human multiple myeloma cell lines and patients SO BRITISH JOURNAL OF HAEMATOLOGY LA English DT Article DE bone sialoprotein; multiple myeloma; bone metastases ID HUMAN BREAST-CANCER AB Bone sialoprotein (BSP) is a glycoprotein essentially found in mineralizing connective tissues, We have recently demonstrated that BSP is ectopically expressed by carcinomas that metastasize to bone with high frequency. Multiple myeloma (MM) is characterized by the localization of tumour plasma cells in the bone marrow. In this study, BSP expression was evaluated in human myeloma cell lines and in bone marrow aspirates and one ascites fluid from MM patients. BSP was detectable in conditioned media of MM cell lines. Using FAGS analysis and in situ hybridization, we demonstrated that tumour cells from all MM patients and cell lines analysed express BSP at both the protein and the mRNA level. C1 Univ Liege, Metastasis Res Lab, B-4000 Liege, Belgium. Univ Liege, Dept Histol, B-4000 Liege, Belgium. NIDCR, Craniofacial & Skeletal Dis Branch, NIH, Bethesda, MD USA. RP Bellahcene, A (reprint author), Univ Liege, Metastasis Res Lab, Tour Pathol 1,Bat B23, B-4000 Liege, Belgium. NR 12 TC 8 Z9 9 U1 0 U2 2 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0007-1048 J9 BRIT J HAEMATOL JI Br. J. Haematol. PD DEC PY 2000 VL 111 IS 4 BP 1118 EP 1121 DI 10.1046/j.1365-2141.2000.02506.x PG 4 WC Hematology SC Hematology GA 399DL UT WOS:000166795900020 PM 11167750 ER PT J AU Rogawski, MA AF Rogawski, MA TI NMDA, AMPA and kainate receptor antagonists as potential antiepileptic drugs SO BRITISH JOURNAL OF PHARMACOLOGY LA English DT Meeting Abstract C1 NINDS, Epilepsy Res Branch, NIH, Bethesda, MD 20892 USA. RI Rogawski, Michael/B-6353-2009 OI Rogawski, Michael/0000-0002-3296-8193 NR 0 TC 0 Z9 0 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0007-1188 J9 BRIT J PHARMACOL JI Br. J. Pharmacol. PD DEC PY 2000 VL 131 SU S MA 225P BP U42 EP U42 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 387FU UT WOS:000166112900073 ER PT J AU Cassedy, JH AF Cassedy, JH TI Mending bodies, saving souls: A history of hospitals. SO BULLETIN OF THE HISTORY OF MEDICINE LA English DT Book Review C1 Natl Lib Med, Bethesda, MD 20894 USA. RP Cassedy, JH (reprint author), Natl Lib Med, Bethesda, MD 20894 USA. NR 1 TC 0 Z9 0 U1 0 U2 0 PU JOHNS HOPKINS UNIV PRESS PI BALTIMORE PA JOURNALS PUBLISHING DIVISION, 2715 NORTH CHARLES ST, BALTIMORE, MD 21218-4319 USA SN 0007-5140 J9 B HIST MED JI Bull. Hist. Med. PD WIN PY 2000 VL 74 IS 4 BP 817 EP 818 DI 10.1353/bhm.2000.0167 PG 2 WC Health Care Sciences & Services; History & Philosophy Of Science SC Health Care Sciences & Services; History & Philosophy of Science GA 392RW UT WOS:000166427100012 ER PT J AU Cantor, D AF Cantor, D TI Gout: The Patrician Malady SO BULLETIN OF THE HISTORY OF MEDICINE LA English DT Book Review C1 NIH, Bethesda, MD 20892 USA. RP Cantor, D (reprint author), NIH, Bethesda, MD 20892 USA. NR 1 TC 0 Z9 0 U1 0 U2 1 PU JOHNS HOPKINS UNIV PRESS PI BALTIMORE PA JOURNALS PUBLISHING DIVISION, 2715 NORTH CHARLES ST, BALTIMORE, MD 21218-4319 USA SN 0007-5140 J9 B HIST MED JI Bull. Hist. Med. PD WIN PY 2000 VL 74 IS 4 BP 820 EP 822 DI 10.1353/bhm.2000.0166 PG 3 WC Health Care Sciences & Services; History & Philosophy Of Science SC Health Care Sciences & Services; History & Philosophy of Science GA 392RW UT WOS:000166427100014 ER PT J AU Cassedy, JH AF Cassedy, JH TI Medicine in Maryland: The Practice and profession, 1799-1999. SO BULLETIN OF THE HISTORY OF MEDICINE LA English DT Book Review C1 Natl Lib Med, Bethesda, MD 20894 USA. RP Cassedy, JH (reprint author), Natl Lib Med, Bethesda, MD 20894 USA. NR 1 TC 0 Z9 0 U1 0 U2 0 PU JOHNS HOPKINS UNIV PRESS PI BALTIMORE PA JOURNALS PUBLISHING DIVISION, 2715 NORTH CHARLES ST, BALTIMORE, MD 21218-4319 USA SN 0007-5140 J9 B HIST MED JI Bull. Hist. Med. PD WIN PY 2000 VL 74 IS 4 BP 834 EP 835 DI 10.1353/bhm.2000.0168 PG 2 WC Health Care Sciences & Services; History & Philosophy Of Science SC Health Care Sciences & Services; History & Philosophy of Science GA 392RW UT WOS:000166427100022 ER PT J AU Zee, RYL Myers, RH Hannan, MT Wilson, PWF Ordovas, JM Schaefer, EJ Lindpaintner, K Kiel, DP AF Zee, RYL Myers, RH Hannan, MT Wilson, PWF Ordovas, JM Schaefer, EJ Lindpaintner, K Kiel, DP TI Absence of linkage for bone mineral density to chromosome 12q12-14 in the region of the vitamin D receptor gene SO CALCIFIED TISSUE INTERNATIONAL LA English DT Article DE VDR; BMD; linkage; Framingham cohort ID FOOD FREQUENCY QUESTIONNAIRE; JAPANESE WOMEN; POLYMORPHISM; OSTEOPOROSIS; ALLELES; REPRODUCIBILITY; ASSOCIATION; POPULATION; VALIDITY; DISEASE AB Polymorphisms in the region of the gene for the vitamin D receptor (VDR) (chromosome 12q12-14) have been associated with differences in bone mineral density (BMD) in some studies but not in others. Because linkage analysis assesses allele sharing identical-by-descent among relatives instead of the association of a particular allele of an anonymous marker. we have performed a linkage study for bone BMD using microsatellite markers flanking the VDR locus. The present study explores whether or not relatives who share the chromosomal region containing the VDR gene have more similar bone density. Participants in the Framingham Osteoporosis Study (aged 37-89 years) who had undergone BMD testing were used to test for concordance of genotype with phenotype in the hip (femoral neck, Ward's area, trochanter) and lumbar spine (L2-L4) with adjustment for covariates. Multipoint quantitative trait link age analysis using variance components methods was conducted with microsatellite markers flanking the VDR locus (GATA91H06, GATA5A09, GGAT2G06) in 332 extended families containing 1062 individuals with both bone density measures and marker data. In addition, quantitative trait sib-pair linkage analysis, with a marker (AFM345xf1) in close proximity to the VDR locus, was performed in a second sample of 169 sibships (n = 413), comprising 284 full-sib pairs. Neither analysis revealed evidence for linkage of this region to femoral neck, Ward's area, lumbar spine, and trochanter in age or sex BMI, and height-adjusted bone density measures. Additional adjustment for alcohol intake, caffeine consumption, smoking status, and estrogen supplement (female only) did not alter the results. The present study could not demonstrate linkage of BMD to chromosome 12q12-14. These findings suggest that neither the VDR gene nor other genes at this locus are likely to have a substantial impact upon bone density. C1 Harvard Univ, Sch Med, Hebrew Rehabil Ctr Aged Res & Training Inst, Boston, MA 02115 USA. Harvard Univ, Sch Med, Div Aging, Boston, MA 02115 USA. Tufts Univ, Jean Mayer US Dept Agr Human Nutr Res Ctr Aging, Lipid Metab Lab, Mol Biol Sect, Boston, MA 02111 USA. NHLBI, Framingham Heart Study, Framingham, MA USA. Boston Univ, Sch Med, Sect Neurogenet, Boston, MA 02118 USA. F Hoffmann La Roche & Co Ltd, Roche Genet, CH-4002 Basel, Switzerland. Max Delbruck Ctr Mol Med, Berlin, Germany. Harvard Univ, Brigham & Womens Hosp, Sch Med, Dept Med,Endocrine Hypertens Div, Boston, MA 02115 USA. RP Kiel, DP (reprint author), Harvard Univ, Sch Med, Hebrew Rehabil Ctr Aged Res & Training Inst, Boston, MA 02115 USA. OI Kiel, Douglas/0000-0001-8474-0310; Ordovas, Jose/0000-0002-7581-5680 FU NIA NIH HHS [AG13645]; NIAMS NIH HHS [AR/AG41398] NR 32 TC 12 Z9 13 U1 1 U2 1 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0171-967X J9 CALCIFIED TISSUE INT JI Calcif. Tissue Int. PD DEC PY 2000 VL 67 IS 6 BP 434 EP 439 DI 10.1007/s002230001175 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 410NK UT WOS:000167445900002 PM 11289690 ER PT J AU Mulshine, JL De Luca, LM Dedrick, RL Tockman, MS Webster, R Placke, ME AF Mulshine, JL De Luca, LM Dedrick, RL Tockman, MS Webster, R Placke, ME TI Considerations in developing successful, population-based molecular screening and prevention of lung cancer SO CANCER LA English DT Article; Proceedings Paper CT International Conference on Prevention and Early Diagnosis of Lung Cancer CY DEC 09-10, 1998 CL VARESE, ITALY SP Amer Canc Soc DE heterogeneous nuclear ribonucleoprotein A2/B1; lung cancer; molecular screening; mortality rate ID CHEMOPREVENTION; RETINOIDS AB The current mortality rate for lung cancer exceeds 85%, as it has for the last 3 decades. This statistic reflects the utility of the major diagnostic tool that has been used during this period to diagnose lung cancer: the chest X-ray. The overwhelming majority of new cases of lung cancer that are detected with chest X-rays involve individuals who already have regional or distant metastatic disease. Because the systemic treatment of this disease has not improved greatly, patients with metastatic disease rarely are cured. This article reviews the issues involved with the development of sputum-based cellular diagnostics for early stage lung cancer. The biomarker, heterogeneous nuclear ribonucleoprotein A2/B1, is the lead marker for this approach. It has been used in several studies in independent cohorts that have suggested that its overexpression in bronchial epithelial cells is associated highly with the development of lung cancer. This marker is detectable 1 year or more prior to the detection of lung cancer by chest X-ray. Finding this early airway-confined phase of lung cancer may allow for the evolution of new management approaches for very early stage lung cancer. Research activities, such aerosolized chemoprevention, are discussed. Cancer 2000;89:2465-7. (C) 2000 American Cancer Society. C1 NCI, Intervent Sect, Med Branch, Bethesda, MD 20892 USA. NIH, Bethesda, MD 20892 USA. USF Moffitt Canc Ctr, Tampa, FL USA. Chiron Diagnost, Emeryville, CA USA. Battelle Mem Inst, Columbus, OH USA. RP Mulshine, JL (reprint author), NCI, Intervent Sect, Med Branch, Bldg 10,12N226,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 20 TC 8 Z9 10 U1 0 U2 3 PU WILEY-BLACKWELL PI MALDEN PA COMMERCE PLACE, 350 MAIN ST, MALDEN 02148, MA USA SN 0008-543X J9 CANCER-AM CANCER SOC JI Cancer PD DEC 1 PY 2000 VL 89 IS 11 SU S BP 2465 EP 2467 PG 3 WC Oncology SC Oncology GA 380QE UT WOS:000165712900024 PM 11147628 ER PT J AU Virtamo, J Edwards, BK Virtanen, M Taylor, PR Malila, N Albanes, D Huttunen, JK Hartman, AM Hietanen, P Maenpaa, H Koss, L Nordling, S Heinonen, OP AF Virtamo, J Edwards, BK Virtanen, M Taylor, PR Malila, N Albanes, D Huttunen, JK Hartman, AM Hietanen, P Maenpaa, H Koss, L Nordling, S Heinonen, OP TI Effects of supplemental alpha-tocopherol and beta-carotene on urinary tract cancer: incidence and mortality in a controlled trial (Finland) SO CANCER CAUSES & CONTROL LA English DT Article DE beta-carotene; chemoprevention; randomized controlled trials; renal cell cancer; urothelial cancer; vitamin E ID SUPERFICIAL BLADDER-TUMORS; RENAL-CELL CARCINOMA; LUNG-CANCER; VITAMIN-E; RETINOL; PREVENTION; ETRETINATE; NUTRITION; DISEASE AB Objectives: Epidemiological studies have suggested a protective effect of vegetables and fruits on urinary tract cancer but the possible protective nutrients are unknown. We studied the effect of alpha-tocopherol (a form of vitamin E) and beta-carotene supplementation on urinary tract cancer in the Alpha-Tocopherol, Beta-Carotene Cancer Prevention (ATBC) Study. Methods: A total of 29,133 male smokers aged 50-69 years from southwestern Finland were randomly assigned to receive alpha-tocopherol (50 mg), beta-carotene (20 mg), both agents, or a placebo daily for 5-8 years (median 6.1 years). Incident urothelial cancers (bladder, ureter, and renal pelvis; n = 169) and renal cell cancers (n = 102) were identified through the nationwide cancer registry. The diagnoses were centrally confirmed by review of medical records and pathology specimens. The supplementation effects were estimated using a proportional hazards model. Results: Neither alpha-tocopherol nor beta-carotene affected the incidence of urothelial cancer, relative risk 1.1 (95% confidence interval (CI) 0.8-1.5) and 1.0 (95% CI 0.7-1.3), respectively, or the incidence of renal cell cancer, relative risk 1.1 (95% CI 0.7-1.6) and 0.8 (95% CI 0.6-1.3), respectively. Conclusion: Long-term supplementation with alpha-tocopherol and beta-carotene has no preventive effect on urinary tract cancers in middle-aged male smokers. C1 Natl Publ Hlth Inst, Dept Nutr, SF-00300 Helsinki, Finland. NCI, Bethesda, MD 20892 USA. Helsinki Univ Hosp, FIN-00170 Helsinki, Finland. Montefiore Med Ctr, Bronx, NY 10467 USA. Univ Helsinki, Helsinki, Finland. RP Virtamo, J (reprint author), Natl Publ Hlth Inst, Dept Nutr, Mannerheimintie 166, SF-00300 Helsinki, Finland. RI Albanes, Demetrius/B-9749-2015 FU NCI NIH HHS [N01-CN-45165] NR 30 TC 56 Z9 56 U1 0 U2 4 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0957-5243 J9 CANCER CAUSE CONTROL JI Cancer Causes Control PD DEC PY 2000 VL 11 IS 10 BP 933 EP 939 DI 10.1023/A:1026546803917 PG 7 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 376ZD UT WOS:000165486400007 PM 11142528 ER PT J AU Hsing, AW Deng, J Sesterhenn, IA Mostofi, FK Stanczyk, FZ Benichou, J Xie, T Gao, YT AF Hsing, AW Deng, J Sesterhenn, IA Mostofi, FK Stanczyk, FZ Benichou, J Xie, T Gao, YT TI Body size and prostate cancer: A population-based case-control study in China SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Article ID UNITED-STATES; MASS INDEX; FAT DISTRIBUTION; RISK-FACTORS; ATTRIBUTABLE RISK; PHYSICAL-ACTIVITY; VISCERAL OBESITY; INSULIN LEVELS; SEX-HORMONES; BIRTH-WEIGHT AB We conducted a population-based case-control study in China to investigate whether body size plays a role in prostate cancer etiology and whether it can explain the rapid increase in prostate cancer incidence rates in China. A total of 238 cases newly diagnosed with primary prostate cancer in Shanghai, China, during 1993-1995 were included in the study. Four hundred and seventy-one healthy control subjects were randomly selected from among residents of Shanghai and frequency-matched to cases on the basis of age. In-person interviews were conducted to elicit information on height, weight history, and other lifestyle factors, Waist and hip circumferences were measured at interview. Odds ratios (ORs) were used to measure the association between prostate cancer and anthropometric variables including height, weight, body mass index (BMI), waist, hip, end right upper arm circumferences, and waist-to-hip ratio (WHR; an indicator of abdominal adiposity), High levels of WHR were related to an excess risk, with men in the highest quartile WKR > 0.92) having an almost 3-fold risk (OR, 2.71; 95% CI = 1.66-4.41; P-trend = 0.0001) compared with men in the lowest quartile (WHR < 0,86), In contrast, men in the highest quartile of hip circumference (>97.4 cm) had a reduced risk (OR, 0.46; 95% CI = 0.29-0.74; P-trend = 0.0002) relative to men in the lowest quartile (<86 cm), No association was found for height, usual adult weight, or preadult and usual adult BMI, Our results suggest that even in a very lean population (average BMI = 21,9), abdominal adiposity may be associated with an increased risk of clinical prostate cancer, pointing to a role of hormones in prostate cancer etiology, Additional research is needed to confirm these findings in prospective studies, especially in Western populations where abdominal obesity is much more common, and to clarify the underlying hormonal mechanisms involved. C1 NCI, Div Canc Epidemiol & Genet, Rockville, MD 20852 USA. Inst Canc Res, Shanghai, Peoples R China. Univ So Calif, Los Angeles, CA 90033 USA. Univ Rouen, F-76031 Rouen, France. Armed Forces Inst Pathol, Washington, DC 20306 USA. Shanghai First Municipal Hosp, Dept Urol, Shanghai 200032, Peoples R China. RP Hsing, AW (reprint author), NCI, Div Canc Epidemiol & Genet, 6120 Execut Blvd,Execut Plaza S,MSC 7234,Room 705, Rockville, MD 20852 USA. EM hsinga@exchange.nih.gov NR 56 TC 102 Z9 104 U1 1 U2 5 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1055-9965 J9 CANCER EPIDEM BIOMAR JI Cancer Epidemiol. Biomarkers Prev. PD DEC PY 2000 VL 9 IS 12 BP 1335 EP 1341 PG 7 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 386EH UT WOS:000166046900009 PM 11142419 ER PT J AU Kimchi-Sarfaty, C Rund, D Ben-Nun-Shaul, O Oppenheim, A Gottesman, MM AF Kimchi-Sarfaty, C Rund, D Ben-Nun-Shaul, O Oppenheim, A Gottesman, MM TI Efficient transfer of human MDR1 cDNA or GFP into hematopoietic cells expressing MHC class I using an SV40 in vitro packaging delivery system SO CANCER GENE THERAPY LA English DT Meeting Abstract C1 NCI, LCB, NIH, Bethesda, MD 20892 USA. Hebrew Univ Jerusalem, Hadassah Med Sch, Hadassah Univ Hosp, IL-91010 Jerusalem, Israel. NR 0 TC 0 Z9 0 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0929-1903 J9 CANCER GENE THER JI Cancer Gene Ther. PD DEC PY 2000 VL 7 IS 12 SU S MA PD49 BP S14 EP S15 PG 2 WC Biotechnology & Applied Microbiology; Oncology; Genetics & Heredity; Medicine, Research & Experimental SC Biotechnology & Applied Microbiology; Oncology; Genetics & Heredity; Research & Experimental Medicine GA 379LK UT WOS:000165643400049 ER PT J AU Shah, AH Kundu, SD Kim, SJ Lee, C AF Shah, AH Kundu, SD Kim, SJ Lee, C TI Modulation of host immunosurveillance of tumors by abrogating TGF-B signaling in lymphocytes SO CANCER GENE THERAPY LA English DT Meeting Abstract C1 Northwestern Univ, Sch Med, Dept Urol, Chicago, IL 60611 USA. NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0929-1903 J9 CANCER GENE THER JI Cancer Gene Ther. PD DEC PY 2000 VL 7 IS 12 SU S MA PD92 BP S28 EP S28 PG 1 WC Biotechnology & Applied Microbiology; Oncology; Genetics & Heredity; Medicine, Research & Experimental SC Biotechnology & Applied Microbiology; Oncology; Genetics & Heredity; Research & Experimental Medicine GA 379LK UT WOS:000165643400091 ER PT J AU Wolf, M El-Rifai, W Tarkkanen, M Kononen, J Serra, M Eriksen, EF Elomaa, I Kallioniemi, A Kallioniemi, OP Knuutila, S AF Wolf, M El-Rifai, W Tarkkanen, M Kononen, J Serra, M Eriksen, EF Elomaa, I Kallioniemi, A Kallioniemi, OP Knuutila, S TI Novel findings in gene expression detected in human osteosarcoma by cDNA microarray SO CANCER GENETICS AND CYTOGENETICS LA English DT Article ID COMPARATIVE GENOMIC HYBRIDIZATION; PROTEIN; GROWTH; CELLS; THROMBOSPONDIN; METASTASES; SARCOMAS; CANCER; TUMOR AB cDNA microarray analysis was used to screen for gene expression alterations in human osteosarcoma cell lines. The analysis using three cell lines revealed changes in the expression of several genes in comparison with normal human osteoblasts. Among the 5,184 sequences that were analyzed, 35 showed aberrant expression in all the cell lines. Eight of these showed overexpression and 27 underexpression compared to their expression levels in osteoblasts. The most highly upregulated genes included heat shock protein 90 beta and polyadenylate-binding protein-like 1. Commonly down-regulated genes included fibronectin 1 and thrombospondin 1. RT-PCR was used to verify these changes in the cell lines and in three primary osteosarcoma samples. This study shows that (1) gene expression pattern in osteosarcoma cell lines differs considerably from normal osteo; blasts, (2) osteosarcoma cell lines can be used as a model system to detect novel gene expression alterations present in primary tumors, (3) the overexpression of heat shock protein 90 beta and poly adenylate-binding protein-like 1, and (4) the down-regulation of fibronectin 1 and thrombospondin 1 may play a role in the development and/or progression of osteosarcoma. This study indicates that microarray-based expression surveys may be used to establish the molecular fingerprint of osteosarcoma, however, larger cDNA chips and more tumor specimens are required to define the clinically relevant gene expression patterns. (C) 2000 Elsevier Science Inc. All rights reserved. C1 Univ Helsinki, Haartman Inst, Dept Med Genet, Helsinki, Finland. Univ Helsinki, Cent Hosp, Dept Oncol, Helsinki, Finland. NHGRI, Canc Genet Lab, NIH, Bethesda, MD 20892 USA. Ist Ortoped Rizzoli, Lab Ric Oncol, Bologna, Italy. Univ Aarhus, Dept Endocrinol, Aarhus, Denmark. Tampere Univ Hosp, Canc Genet Lab, Tampere, Finland. Univ Tampere, Inst Med Technol, FIN-33101 Tampere, Finland. RP Knuutila, S (reprint author), Univ Helsinki, Haartman Inst, Dept Med Genet, Helsinki, Finland. RI Kallioniemi, Olli/H-5111-2011; Kallioniemi, Olli/H-4738-2012; Serra, Massimo/J-4878-2016; OI Kallioniemi, Olli/0000-0002-3231-0332; Kallioniemi, Olli/0000-0002-3231-0332; Serra, Massimo/0000-0003-0742-1177; Kallioniemi, Anne/0000-0003-3552-8158 NR 18 TC 42 Z9 47 U1 0 U2 3 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0165-4608 J9 CANCER GENET CYTOGEN JI Cancer Genet. Cytogenet. PD DEC PY 2000 VL 123 IS 2 BP 128 EP 132 DI 10.1016/S0165-4608(00)00319-8 PG 5 WC Oncology; Genetics & Heredity SC Oncology; Genetics & Heredity GA 385JM UT WOS:000166000700006 PM 11156738 ER PT J AU Arlen, P Tsang, KY Marshall, JL Chen, A Steinberg, SM Poole, D Hand, PH Schlom, J Hamilton, JM AF Arlen, P Tsang, KY Marshall, JL Chen, A Steinberg, SM Poole, D Hand, PH Schlom, J Hamilton, JM TI The use of a rapid ELISPOT assay to analyze peptide-specific immune responses in carcinoma patients to peptide vs. recombinant poxvirus vaccines SO CANCER IMMUNOLOGY IMMUNOTHERAPY LA English DT Article DE avipox-CEA; ELISPOT assay; CEA peptide; immune responses; cancer vaccine; carcinoma ID HUMAN CARCINOEMBRYONIC ANTIGEN; COLORECTAL-CANCER PATIENTS; CYTOTOXIC T-CELLS; DENDRITIC CELLS; ACTIVE IMMUNOTHERAPY; MELANOMA PATIENTS; PERIPHERAL-BLOOD; TRANSGENIC MICE; FLOW-CYTOMETRY; FUSION PROTEIN AB An enzyme-linked immunosorbent spot (ELISPOT) assay for interferon gamma production has been used to analyze specific T cell responses to a Flu 9-mer peptide, and a 9-mer peptide of carcinoembryonic antigen (CEA). Assays were performed on peripheral blood mononuclear cells (PBMC) of HLA-A2-positive patients with CEA-expressing carcinomas, both before and after vaccination with CEA-based vaccines, and from HLA-A2-positive healthy blood donors. The ELISPOT assay utilized aliquots of frozen PBMC, and assays were performed after 24 h in culture with peptide to rule out any artifacts due to long-term in vitro stimulation cycles. An internal standard was used for each assay to define reproducibility of the assay, and all samples from a given patient (pre- and post-vaccination, with both the Flu and CEA peptides) were analyzed simultaneously. The results indicated a trend towards healthy blood donors having higher levels of Flu-specific T cell precursors than do colon carcinoma patients, but these results were not statistically significant (P = 0.06). On the other hand, slightly higher CEA-specific T cell responses were observed in cancer patients with CEA-expressing carcinomas than in healthy blood donors. PBMC from two CEA-based vaccine clinical trials were analyzed for T cell responses to the same CEA peptide and to the Flu control peptide. The first trial consisted of three monthly vaccinations of CEA peptide (Designated PPP) in adjuvant. The second trial consisted of cohorts receiving three monthly vaccinations of avipox-CEA recombinant (designated AAA) or cohorts receiving a primary vaccination with recombinant vaccinia-CEA followed by two monthly vaccinations with avipox-CEA (designated VAA). Few, if any, CEA-specific T cell responses were seen in the PPP vaccinations, while the majority of patients receiving the poxvirus CEA recombinants demonstrated increases in CEA-specific T cell responses and no increases in Flu-specific responses. CEA-specific IgG responses were also demonstrated in patients following recombinant CEA poxvirus vaccinations. Statistical analyses of the T cell responses to the same CEA peptide demonstrated a P value of 0.028 for the recombinant poxvirus vaccines, as compared with the peptide vaccine. There were no differences seen (P = 0.37) in Flu-specific responses after these two types of CEA vaccination. These results thus provide the first evidence that poxvirus recombinant-based vaccines are more potent in the initiation of tumor-antigen-specific T cell responses than vaccines employing peptide in adjuvant, when assays are conducted in an identical manner, and in defining responses to the same peptide. These results also demonstrate for the first time that an ELISPOT assay, performed over a 24-h period and without in vitro sensitization, can be successfully used to monitor immune responses to a tumor-associated antigen in cancer patients. C1 NCI, Tumor Immunol & Biol Lab, NIH, Bethesda, MD 20892 USA. Georgetown Univ, Med Ctr, Vincent T Lombardi Canc Res Ctr, Washington, DC 20007 USA. NCI, Med Branch, NIH, Bethesda, MD 20892 USA. NCI, Biostat & Data Management Sect, NIH, Bethesda, MD 20892 USA. RP Schlom, J (reprint author), NCI, Tumor Immunol & Biol Lab, NIH, 10 Ctr Dr, Bethesda, MD 20892 USA. NR 76 TC 61 Z9 65 U1 0 U2 2 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0340-7004 J9 CANCER IMMUNOL IMMUN JI Cancer Immunol. Immunother. PD DEC PY 2000 VL 49 IS 10 BP 517 EP 529 DI 10.1007/s002620000145 PG 13 WC Oncology; Immunology SC Oncology; Immunology GA 380FA UT WOS:000165688000001 PM 11129322 ER PT J AU Urasaki, Y Takebayashi, Y Pommier, Y AF Urasaki, Y Takebayashi, Y Pommier, Y TI Activity of a novel camptothecin analogue, homocamptothecin, in camptothecin-resistant cell lines with topoisomerase I alterations SO CANCER RESEARCH LA English DT Article ID RING-MODIFIED CAMPTOTHECIN; DNA; STABILITY; MUTATION; CEM/C2 AB Homocamptothecin (hCPT), which differs from camptothecin (CPT) by the presence of an additional methylene group in the E-ring, was evaluated in CPT-resistant cell lines. Topoisomerase I (top1)-deficient leukemia P388/CPT45 cells were highly resistant to hCPT, which demonstrates that top1 is the primary target of hCPT, Three CPT-resistant cell lines with top1 point mutations (Chinese hamster lung fibroblast DC3F/C10, human prostate carcinoma DU-145/RC1, and human leukemia CEM/C2) and their top1 enzymes were cross-resistant to hCPT, The antiproliferative activity of hCPT was greater than that of CPT in both parental and CPT-resistant cell lines, particularly in the prostate cell lines. The top1 cleavage complexes formed in the presence of hCPT appear to be more stable than those induced by CPT. Together, these data indicate that hCPT is a specific top1 inhibitor, which shares a common binding site with CPT in the top1-DNA cleavage complexes. Because of its potency, hCPT might overcome resistance to CPT in some cancer cells. C1 NCI, Mol Pharmacol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. Tohoku Univ, Inst Dev Aging & Canc, Dept Pathol, Sendai, Miyagi 9808575, Japan. RP Pommier, Y (reprint author), NCI, Mol Pharmacol Lab, Div Basic Sci, NIH, Bldg 37,Room 4E28,37 Convent Dr,MSC 4255, Bethesda, MD 20892 USA. NR 18 TC 28 Z9 32 U1 1 U2 2 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD DEC 1 PY 2000 VL 60 IS 23 BP 6577 EP 6580 PG 4 WC Oncology SC Oncology GA 379JJ UT WOS:000165638700006 PM 11118036 ER PT J AU Sallinen, SL Sallinen, PK Haapasalo, HK Helin, HJ Helen, PT Schraml, P Kallioniemi, OP Kononen, J AF Sallinen, SL Sallinen, PK Haapasalo, HK Helin, HJ Helen, PT Schraml, P Kallioniemi, OP Kononen, J TI Identification of differentially expressed genes in human gliomas by DNA microarray and tissue chip techniques SO CANCER RESEARCH LA English DT Article ID CDNA MICROARRAYS; GROWTH-FACTOR; PATTERNS AB New genomic large-scale screening techniques have made the task of establishing an accurate molecular fingerprint of cancer cells feasible. Here, we have used a two-phase strategy for identification of molecular alterations in gliomas, First, cDNA microarrays (Clontech Laboratories, Inc., Research Genetics) were used to pinpoint differentially expressed genes between normal brain and diffuse astrocytomas (grades II-IV), and between a primary tumor and a later tumor reoccurrence in the same patient, More than 200 gene expression alterations were detected from glioblastomas, whereas relatively few changes were seen in grade II and grade III tumors. The most distinct progression-related expression change was the up-regulation of the insulin-like growth factor binding protein 2 (IGFBP2) gene. Second, a high-density tissue microarray of 418 brain tumors was constructed and used for clinical validation of gene expression changes. Strong expression of IGFBP2 was associated with progression and poor patient survival in diffuse astrocytomas (P < 0.0001). Third, comparisons of the data between (a) multiple spots retrieved from one predefined tumor region (IGFBP2 and vimentin immunohistochemistry, 20 tumors) or between (b) standard slides and arrayed tissues (p53 immunohistochemistry, 42 tumors) revealed very little variation. In conclusion, the combined use of DNA microarrays and tissue microarrays offers a powerful strategy for rapid identification and thorough characterization of differentially expressed genes in gliomas. C1 NHGRI, Canc Genet Lab, NIH, Bethesda, MD 20892 USA. Tampere Univ Hosp, Dept Pathol, FIN-33521 Tampere, Finland. Tampere Univ Hosp, Lab Mol Pathol, FIN-33521 Tampere, Finland. Tampere Univ Hosp, Dept Neurosurg, FIN-33521 Tampere, Finland. Univ Basel, Inst Pathol, CH-4003 Basel, Switzerland. RP Kononen, J (reprint author), NHGRI, Canc Genet Lab, NIH, 49 Convent Dr,MSC 4470,Room 4A24, Bethesda, MD 20892 USA. RI Kallioniemi, Olli/H-5111-2011; Kallioniemi, Olli/H-4738-2012 OI Kallioniemi, Olli/0000-0002-3231-0332; Kallioniemi, Olli/0000-0002-3231-0332 NR 13 TC 257 Z9 286 U1 3 U2 14 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD DEC 1 PY 2000 VL 60 IS 23 BP 6617 EP 6622 PG 6 WC Oncology SC Oncology GA 379JJ UT WOS:000165638700014 PM 11118044 ER PT J AU Rusyn, I Denissenko, MF Wong, VA Butterworth, BE Cunningham, ML Upton, PB Thurman, RG Swenberg, JA AF Rusyn, I Denissenko, MF Wong, VA Butterworth, BE Cunningham, ML Upton, PB Thurman, RG Swenberg, JA TI Expression of base excision repair enzymes in rat and mouse liver is induced by peroxisome proliferators and is dependent upon carcinogenic potency SO CARCINOGENESIS LA English DT Article ID DNA-POLYMERASE-BETA; MESSENGER-RNA; KAPPA-B; DAMAGE; DRUGS; 8-HYDROXYDEOXYGUANOSINE; OVEREXPRESSION; SENSITIVITY; GLYCOSYLASE; ACTIVATION AB Elevated and sustained cell replication, together with a decrease in apoptosis, is considered to be the main mechanism of hepatic tumor promotion due to peroxisome proliferators, In contrast, the role of oxidative stress and DNA damage in the carcinogenic mechanism is less well understood. In view of possible induction of DNA damage by peroxisome proliferators, DNA repair mechanisms may be an important factor to consider in the mechanism of action of these compounds. Here, the ability of peroxisome proliferators to induce expression of base excision repair enzymes was examined. WY-14,643, a potent carcinogen, increased expression of several base excision DNA repair enzymes in a dose- and time-dependent manner. Importantly, expression of enzymes that do not repair oxidative DNA damage was not changed, Moreover, less potent members of the peroxisome proliferator group had much weaker or no effects on expression of DNA repair enzymes when compared with WY-14,643. Collectively, these data suggest that DNA base excision repair may be an important factor in peroxisome proliferator-induced carcinogenesis and that induction of DNA repair might provide further evidence supporting a role of oxidative DNA damage by peroxisome proliferators. C1 Univ N Carolina, Dept Environm Sci & Engn, Chapel Hill, NC 27599 USA. Univ N Carolina, Curriculum Toxicol, Chapel Hill, NC 27599 USA. Univ N Carolina, Dept Pharmacol, Chapel Hill, NC 27599 USA. BD PharMingen, San Diego, CA 92121 USA. Chem Ind Inst Toxicol, Res Triangle Pk, NC 27709 USA. NIEHS, NIH, Res Triangle Pk, NC 27709 USA. RP Swenberg, JA (reprint author), Univ N Carolina, Dept Environm Sci & Engn, CB 7400, Chapel Hill, NC 27599 USA. RI Rusyn, Ivan/S-2426-2016 FU NIEHS NIH HHS [ES-09785] NR 32 TC 46 Z9 47 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0143-3334 J9 CARCINOGENESIS JI Carcinogenesis PD DEC PY 2000 VL 21 IS 12 BP 2141 EP 2145 DI 10.1093/carcin/21.12.2141 PG 5 WC Oncology SC Oncology GA 391FZ UT WOS:000166347100001 PM 11133801 ER PT J AU Kiss, A Aguilera, G AF Kiss, A Aguilera, G TI Role of alpha-1-adrenergic receptors in the regulation of corticotropin-releasing hormone mRNA in the paraventricular nucleus of the hypothalamus during stress SO CELLULAR AND MOLECULAR NEUROBIOLOGY LA English DT Article DE alpha-adrenergic; corticotropin-releasing hormone; HPA axis; hypothalamic paraventricular nucleus; stress ID ADRENOCORTICOTROPIN SECRETION; RAT HYPOTHALAMUS; ANGIOTENSIN-II; NEURONS; VASOPRESSIN; NOREPINEPHRINE; CATECHOLAMINES; CORTICOSTERONE; ACTIVATION; EXPRESSION AB 1. The role of alpha1-adrenergic receptors on CRH mRNA levels in the PVN was studied in control and stressed rats receiving i.c.v. injections of the alpha1-adrenergic agonist, methoxamine, or the alpha1- antagonist, prazosin. 2. Plasma ACTH increased significantly 60 min and 4 hr after a single injection of methoxamine (100 mug, i.c.v.). No desensitization of this response was observed after repeated injections every 6 hr for 24 hr. Concomitantly, POMC mRNA in the anterior pituitary increased by 25% at 4 hr after a single injection and by 96% after repeated injections. 3. CRH mRNA levels in the PVN increased by 131% after repeated injections for 24 hr, but were unchanged 4 hr after a single injection. Central alpha1-adrenergic blockade with prazosin did not prevent the increases in CRH mRNA following 4 hr of acute stress, but significantly reduced the increases observed 24 hr after an i.c.v. injection of 75 mug of colchicine or after repeated i.p. hypertonic saline injections every 8 hr. 4. These studies demonstrate that while alpha1-adrenergic receptors contribute to longterm increases of CRH mRNA levels in the PVN during prolonged stress, other factors are likely to be involved in the stimulation of CRH mRNA following acute stimulation. C1 NICHD, Dev Endocrinol Branch, Sect Endocrine Physiol, NIH, Bethesda, MD 20892 USA. RP Aguilera, G (reprint author), NICHD, Dev Endocrinol Branch, Sect Endocrine Physiol, NIH, Bldg 10,Rm 10N262,9000 Rockville Pike, Bethesda, MD 20892 USA. EM greti@helix.nih.gov NR 38 TC 28 Z9 28 U1 0 U2 3 PU SPRINGER/PLENUM PUBLISHERS PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 0272-4340 J9 CELL MOL NEUROBIOL JI Cell. Mol. Neurobiol. PD DEC PY 2000 VL 20 IS 6 BP 683 EP 694 DI 10.1023/A:1007098724683 PG 12 WC Cell Biology; Neurosciences SC Cell Biology; Neurosciences & Neurology GA 376FE UT WOS:000165446900006 PM 11100976 ER PT J AU Borinstein, SC Hyatt, MA Sykes, VW Straub, RE Lipkowitz, S Boulter, J Bogler, O AF Borinstein, SC Hyatt, MA Sykes, VW Straub, RE Lipkowitz, S Boulter, J Bogler, O TI SETA is a multifunctional adapter protein with three SH3 domains that binds Grb2, Cbl, and the novel SB1 proteins SO CELLULAR SIGNALLING LA English DT Article DE SH3; adapter; glioma; Grb2; Cbl; astrocyte; AIP1; ALG-2; apoptosis ID GROWTH-FACTOR RECEPTOR; C-CBL; COILED COILS; DRUG DESIGN; CELLS; PROTOONCOGENE; REGULATOR; APOPTOSIS; ALG-2; EXPRESSION AB Expression of the src homology 3 (SH3)-encoding, expressed in tumorigenic astrocytes (SETA) gene is associated with astrocyte transformation in culture and tumors in the adult brain. SETA binds to the apoptosis regulator apoptosis-linked gene 2 (ALG-2) interacting protein 1 (AIP I), and modulates apoptosis in astrocytes. The predicted protein structure of SETA revealed two SH3 domains, while related proteins were reported to have three. Here we report the identification of an additional SH3 domain N-terminal to the previously identified SETA sequence. Yeast two-hybrid screening of a p53(-/-) astrocyte cDNA library with this; SH3 domain identified a novel gene, SETA binding protein I (SB1), with 55% amino acid identity to the renal tumor antigen, NY-REN-45. In vitro confrontation and coimmunoprecipitation experiments confirmed the binding of SB1 to SETA. Evidence that SETA binds to the CD2 protein, the proto-oncogene c-Cbl, and the signal transduction molecule Grb2, and can dimerize via its C-terminal coiled coil (CC) domain is also presented. (C) 2000 Elsevier Science Inc. All rights reserved. C1 Virginia Commonwealth Univ, Med Coll Virginia, Dept Anat, Richmond, VA 23298 USA. Virginia Commonwealth Univ, Med Coll Virginia, Dept Psychiat, Richmond, VA 23298 USA. NCI, Dept Genet, Med Branch, Div Clin Sci, Bethesda, MD 20892 USA. Uniformed Serv Univ Hlth Sci, Cell Biol Program, Bethesda, MD 20889 USA. Univ Calif Los Angeles, Dept Psychiat & Biobehav Sci, Los Angeles, CA 90024 USA. RP Bogler, O (reprint author), Henry Ford Hosp, Dept Neurosurg, Room 3096,E&R Bldg,2799 W Grand Blvd, Detroit, MI 48202 USA. OI Bogler, Oliver/0000-0002-3700-0480 FU NIDA NIH HHS [P50 DA005010] NR 33 TC 52 Z9 57 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0898-6568 J9 CELL SIGNAL JI Cell. Signal. PD DEC PY 2000 VL 12 IS 11-12 BP 769 EP 779 DI 10.1016/S0898-6568(00)00129-7 PG 11 WC Cell Biology SC Cell Biology GA 386EZ UT WOS:000166048900009 PM 11152963 ER PT J AU King, RS Sharma, V Pedersen, LC Kakuta, Y Negishi, M Duffel, MW AF King, RS Sharma, V Pedersen, LC Kakuta, Y Negishi, M Duffel, MW TI Structure-function modeling of the interactions of N-alkyl-N-hydroxyanilines with rat hepatic aryl sulfotransferase TV SO CHEMICAL RESEARCH IN TOXICOLOGY LA English DT Article ID TYROSINE-ESTER SULFOTRANSFERASE; ESTROGEN SULFOTRANSFERASE; HYDROXY ARYLAMINES; CRYSTAL-STRUCTURE; AMINE OXIDASE; IV; LIVER; N-HYDROXY-2-ACETYLAMINOFLUORENE; SULFATION; MECHANISM AB Although previous investigations have clearly shown that N-hydroxy arylamines and N-hydroxy heterocyclic amines are substrates for sulfotransferases, relatively little is known about which structural features of the N-hydroxy arylamines are important for sulfation to occur. The purpose of this investigation was to determine the extent to which secondary N-alkyl-N-hydroxy arylamines interact with aryl sulfotransferase (AST) IV (also known as tyrosine-ester sulfotransferase or ST1A1) and to evaluate these interactions using molecular modeling techniques. AST IV is a major cytosolic sulfotransferase in the rat, and it catalyzes the sulfation of various phenols, benzylic alcohols, arylhydroxamic acids, oximes, and primary N-hydroxy arylamines. In this study, three secondary N-hydroxy arylamines, N-hydroxy-N-methylaniline, N-ethyl-N-hydroxyaniline, and N-hydroxy-N-n-propylaniline, were found to be substrates for the purified rat hepatic AST IV. However, when the N-alkyl substituent was an n-butyl group (i.e., N-n-butyl-N-hydroxyaniline), the interaction with the enzyme changed from that of a substrate to competitive inhibition. This change in specificity was further explored through the construction and use of a model for AST IV based on mouse estrogen sulfotransferase, an enzyme whose crystal structure has been previously determined to high resolution. Molecular modeling techniques were used to dock each of the above N-hydroxy arylamines into the active site of the homology model of AST IV and determine optimum ligand geometries. The results of these experiments indicated that steric constraints on the orientation of binding of secondary N-alkyl-N-hydroxy arylamines at the active site of AST TV play a significant role in determining the nature of the interaction of the enzyme with these compounds. C1 Univ Iowa, Coll Pharm, Div Med & Nat Prod Chem, Iowa City, IA 52242 USA. Univ Rhode Isl, Coll Pharm, Dept Biomed Sci, Kingston, RI 02881 USA. NIEHS, Pharmacogenet Sect, Reprod & Dev Toxicol Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Duffel, MW (reprint author), Univ Iowa, Coll Pharm, Div Med & Nat Prod Chem, Iowa City, IA 52242 USA. RI King, Roberta/A-1749-2010; Duffel, Michael/E-6066-2015 OI King, Roberta/0000-0002-3550-4255; Duffel, Michael/0000-0002-6079-7459 FU NCI NIH HHS [CA38683, R01 CA038683]; NIGMS NIH HHS [GM07069] NR 38 TC 7 Z9 8 U1 0 U2 1 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0893-228X J9 CHEM RES TOXICOL JI Chem. Res. Toxicol. PD DEC PY 2000 VL 13 IS 12 BP 1251 EP 1258 DI 10.1021/tx990184z PG 8 WC Chemistry, Medicinal; Chemistry, Multidisciplinary; Toxicology SC Pharmacology & Pharmacy; Chemistry; Toxicology GA 386EX UT WOS:000166048600009 PM 11123966 ER PT J AU Harsch, A Sayer, JM Jerina, DM Vouros, F AF Harsch, A Sayer, JM Jerina, DM Vouros, F TI HPLC-MS/MS identification of positionally isomeric benzo[c]phenanthrene diol epoxide adducts in duplex DNA SO CHEMICAL RESEARCH IN TOXICOLOGY LA English DT Article ID MASS-SPECTROMETRY; NUCLEOSIDE ADDUCTS; OLIGONUCLEOTIDES; SEQUENCE; OLIGODEOXYRIBONUCLEOTIDES; MUTATION; BINDING; REPAIR AB LC-MS and LC-MS/MS analyses were used to investigate the chemoselectivity of the carcinogenic dial epoxide metabolite, (-)-(1R,2S,3S,4R)- 1,2-epoxy-3,4-dihydroxy-1,2,3,4-tetrahydrobenzo[c]phenanthrene [(-)-(R,S,S,R)-BcPh DE-2], on reaction in vitro with an oligonucleotide dodecamer derived from the HPRT gene. The sequence of this dodecamer, 5'-T(1)A(2)G(3)T(4)C(5)A(6)A(7)G(8)G(9)G(10)C(11)A(12)-3', contains a base (corresponding to A(7)) which is a hot spot for mutagenesis in the hprt gene induced by the carcinogenic (R,S,S,R)-enantiomer of benzo[alpha ]pyrene 7,8-diol 9,10-epoxide, and an adjacent base (corresponding to As) which gave no mutations with this diol epoxide. Modified oligonucleotides were generated by reaction of(-)BcPh DE-2 with both the single-stranded and duplex forms of the dodecamer. Multiple purine targets in both strands led to the formation of complex reaction mixtures of regioisomeric BcPh DE-modified oligonucleotides, which were partially separated by reverse phase HPLC on a polystyrene-divinylbenzene column. On-line LC-MS data allowed facile distinction between adducts on the two strands of the duplex, and MS/MS analysis permitted unambiguous assignment of the major sites of modification in the regioisomeric, adducted strands. In the duplex, these sites were at A(6), A(7), and G(8). Interestingly, the "hot spot" A(7W) as about 3 times more reactive with the BcPh DE than the "cold spot" A(6). Adduct formation from the single-stranded dodecamer was less selective, and resulted in more extensive alkylation of G residues. C1 NIDDKD, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. Northeastern Univ, Dept Chem, Boston, MA 02115 USA. Northeastern Univ, Barnett Inst, Boston, MA 02115 USA. RP Jerina, DM (reprint author), NIDDKD, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. FU NCI NIH HHS [CA69390] NR 22 TC 26 Z9 27 U1 1 U2 5 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0893-228X J9 CHEM RES TOXICOL JI Chem. Res. Toxicol. PD DEC PY 2000 VL 13 IS 12 BP 1342 EP 1348 DI 10.1021/tx000140m PG 7 WC Chemistry, Medicinal; Chemistry, Multidisciplinary; Toxicology SC Pharmacology & Pharmacy; Chemistry; Toxicology GA 386EX UT WOS:000166048600020 PM 11123977 ER PT J AU Orbach, Y Lamb, ME AF Orbach, Y Lamb, ME TI Enhancing children's narratives in investigative interviews SO CHILD ABUSE & NEGLECT LA English DT Article DE investigative interviews; narrative enhancement; open-ended questions; recall memory ID SEXUAL-ABUSE VICTIMS; EYEWITNESS TESTIMONY; RETENTION INTERVAL; UTTERANCE TYPES; WITNESSES; MEMORY; QUESTION; EVENT; SUGGESTIBILITY; PRESCHOOLERS AB Objective: To illustrate the amount of detail that can be elicited from alleged abuse victims using open-ended prompts by closely examining forensic interviews of a 5-year-old and a 15-year-old. Method: Interview prompts in the substantive sections of two forensic interviews were characterized as invitations, cued invitations, directive or option-posing, and the number of details they each elicited was tabulated. Results: In both interviews, open-ended prompts predominated and were distributed throughout the substantive phases of the interviews. Most of the information obtained was elicited using open-ended prompts, which remained equivalently effective throughout the interviews. Reconstruction of the children's accounts illustrated how successive prompts continued to elicit information. Conclusion: Well-framed open-ended prompts, including those that use details provided by the child as cues, elicit narrative accounts from children of all ages. Because such information is more likely to be accurate, investigators are urged to rely more extensively on open-ended prompts. (C) 2000 Elsevier Science Ltd. C1 NICHHD, Sect Social & Emot Dev, Bethesda, MD 20892 USA. RP Lamb, ME (reprint author), NICHHD, Sect Social & Emot Dev, 9190 Rockville Pike, Bethesda, MD 20892 USA. NR 64 TC 39 Z9 40 U1 1 U2 3 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0145-2134 J9 CHILD ABUSE NEGLECT JI Child Abuse Negl. PD DEC PY 2000 VL 24 IS 12 BP 1631 EP 1648 DI 10.1016/S0145-2134(00)00207-6 PG 18 WC Family Studies; Psychology, Social; Social Work SC Family Studies; Psychology; Social Work GA 434GG UT WOS:000168806300012 PM 11197041 ER PT J AU Nelson, DL Terhorst, C AF Nelson, DL Terhorst, C TI X-linked lymphoproliferative syndrome SO CLINICAL AND EXPERIMENTAL IMMUNOLOGY LA English DT Editorial Material ID BONE-MARROW TRANSPLANTATION; INFECTIOUS-MONONUCLEOSIS; DISEASE; GENE; FEATURES; LYMPHOMA; XLP C1 NCI, Metab Branch, Div Clin Sci, NIH, Bethesda, MD 20892 USA. Harvard Univ, Sch Med, Beth Israel Deaconess Med Ctr, Div Immunol, Boston, MA USA. RP Nelson, DL (reprint author), NCI, Metab Branch, Div Clin Sci, NIH, Bldg 10,Rm 4N115, Bethesda, MD 20892 USA. NR 17 TC 8 Z9 9 U1 0 U2 1 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0009-9104 J9 CLIN EXP IMMUNOL JI Clin. Exp. Immunol. PD DEC PY 2000 VL 122 IS 3 BP 291 EP 295 DI 10.1046/j.1365-2249.2000.01400.x PG 5 WC Immunology SC Immunology GA 377XW UT WOS:000165552400001 PM 11122230 ER PT J AU Moretti, S Marcellini, S Boschini, A Famularo, G Santini, G Alesse, E Steinberg, SM Cifone, MG Kroemer, G De Simone, C AF Moretti, S Marcellini, S Boschini, A Famularo, G Santini, G Alesse, E Steinberg, SM Cifone, MG Kroemer, G De Simone, C TI Apoptosis and apoptosis-associated perturbations of peripheral blood lymphocytes during HIV infection: comparison between AIDS patients and asymptomatic long-term non-progressors SO CLINICAL AND EXPERIMENTAL IMMUNOLOGY LA English DT Article DE long-term non-progressors; lymphocyte apoptosis; mitochondria ID FAS-MEDIATED APOPTOSIS; PROGRAMMED CELL-DEATH; FLOW-CYTOMETRY; T-CELLS; ACIDIC SPHINGOMYELINASE; ANTIRETROVIRAL THERAPY; VIRUS-REPLICATION; MOUSE-LIVER; ICE-LIKE; ACTIVATION AB This study was designed to compare the degree of lymphocyte apoptosis and Fas-Fas ligand (FasL) expression in AIDS patients and long-term non-progressors (LTNPs) and correlate these parameters with apoptosis-associated perturbations in lymphocyte function. LTNPs had a lower frequency of apoptotic CD4(+) and CD8(+) T cells compared with subjects with AIDS. This correlated with a lower frequency of cells expressing Fas and FasL. The frequency of selected lymphocyte populations exhibiting a disrupted mitochondrial transmembrane potential (Delta Psi (m)) and increased superoxide generation was lower in LTNPs than in patients with AIDS; these abnormalities were associated with lower levels of caspase-1 activation in LTNPs. The results indicate a significantly reduced level of apoptosis and apoptosis-associated parameters in LTNPs than in patients developing AIDS. Based on these findings, a crucial role for mitochondria can be predicted in the process of lymphocyte apoptosis during the evolution of AIDS. C1 Univ Aquila, Dept Expt Med, I-67100 Laquila, Italy. Osped San Camillo, Dept Emergency Med, Rome, Italy. NCI, NIH, Bethesda, MD 20892 USA. CNRS, Villejuif, France. RP De Simone, C (reprint author), Univ Aquila, Dept Expt Med, Via Vetoio 10,Coppito 2, I-67100 Laquila, Italy. RI Kroemer, Guido/B-4263-2013; OI CIFONE, Maria Grazia/0000-0002-9923-5445 NR 49 TC 42 Z9 42 U1 0 U2 1 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0009-9104 J9 CLIN EXP IMMUNOL JI Clin. Exp. Immunol. PD DEC PY 2000 VL 122 IS 3 BP 364 EP 373 DI 10.1046/j.1365-2249.2000.01375.x PG 10 WC Immunology SC Immunology GA 377XW UT WOS:000165552400013 PM 11122242 ER PT J AU Feldman, AL Tamarkin, L Paciotti, GF Simpson, BW Linehan, WM Yang, JC Fogler, WE Turner, EM Alexander, HR Libutti, SK AF Feldman, AL Tamarkin, L Paciotti, GF Simpson, BW Linehan, WM Yang, JC Fogler, WE Turner, EM Alexander, HR Libutti, SK TI Serum endostatin levels are elevated and correlate with serum vascular endothelial growth factor levels in patients with stage IV clear cell renal cancer SO CLINICAL CANCER RESEARCH LA English DT Article ID BASEMENT-MEMBRANE ZONES; TUMOR-GROWTH; IN-VITRO; ANGIOGENESIS; EXPRESSION; CARCINOMA; INHIBITOR; COLLAGEN; METASTASES; CYTOKINES AB Clear cell renal carcinoma (CCRC) is a highly angiogenic tumor known to secrete vascular endothelial cell growth factor (VEGF), Endostatin is an endogenous antiangiogenic agent with antitumor activity in mice. The purpose of this study was to evaluate serum levels of endostatin in normal subjects and in patients with CCRC and to examine the relationship of these levels to circulating VEGF levels. Fifteen patients (mean age, 48 years) on a clinical protocol for stage IV CCRC at the National Cancer Institute were included in the study. Archived prenephrectomy serum samples were analyzed for endostatin and VEGF concentrations. Endostatin and VEGF levels were compared with those of an age-matched group of volunteer blood donors (n = 18) using a competitive enzyme immunoassay, Data were analyzed using the Mann-Whitney U test and the Spearman rank correlation, Median serum endostatin levels were 24.6 ng/ml (range, 15.1-54.0 ng/ml) in CCRC patients versus 14.1 ng/ml (range, 1.0-19.3 ng/ml) in healthy controls (P < 0.0001). Median VEGF levels were 3.4 ng/ml (range, 0.1-11.2 ng/ml) and 2.5 ng/ml (range, 0.1-4.2 ng/ml), respectively (P = 0.065). A highly significant correlation was observed between endostatin and VEGF levels among the CCRC patients (r = 0.81, P = 0.0003) but not among controls (r = -0.22, P = 0.37). Endostatin levels are detectable in serum from healthy subjects as well as from CCRC patients, Levels are significantly elevated and correlate with VEGF levels in CCRC patients. Elucidating the nature of this correlation may lend insight into the regulation of tumor angiogenesis in patients with renal cancer. C1 NCI, Surg Branch, NIH, Bethesda, MD 20892 USA. NCI, Urol Oncol Branch, Bethesda, MD 20892 USA. CytImmune Sci Inc, College Pk, MD USA. EntreMed Inc, Rockville, MD USA. RP Libutti, SK (reprint author), NCI, Surg Branch, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. RI Feldman, Andrew/D-5028-2012 NR 36 TC 96 Z9 110 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD DEC PY 2000 VL 6 IS 12 BP 4628 EP 4634 PG 7 WC Oncology SC Oncology GA 388CQ UT WOS:000166160200006 PM 11156212 ER PT J AU Marrogi, AJ Travis, WD Welsh, JA Khan, MA Rahim, H Tazelaar, H Pairolero, P Trastek, V Jett, J Caporaso, NE Liotta, LA Harris, CC AF Marrogi, AJ Travis, WD Welsh, JA Khan, MA Rahim, H Tazelaar, H Pairolero, P Trastek, V Jett, J Caporaso, NE Liotta, LA Harris, CC TI Nitric oxide synthase, cyclooxygenase 2, and vascular endothelial growth factor in the angiogenesis of non-small cell lung carcinoma SO CLINICAL CANCER RESEARCH LA English DT Article ID FACTOR EXPRESSION; GENE-EXPRESSION; P53 MUTATIONS; TUMOR-GROWTH; CANCER; PROGNOSIS; COLON; VEGF; THROMBOSPONDIN-1; ADENOCARCINOMA AB We have investigated the hypothesis that nitric oxide synthase (NOS2), cyclooxygenase-2 (COX2), and vascular endothelial growth factor (VEGF) protein levels individually demonstrate a direct correlation with microvessel density (MVD) and clinical outcome in human non-small cell lung cancer (NSCLC), Furthermore, we hypothesized that MVD may explain the propensity of certain histological lung cancer subtypes for early metastasis via a hematological route, Immunohistochemically, we studied the protein expression levels of NOS2, COX2, and VEGF and MVD by counting: CD31-reactive blood vessels (BVs) in 106 surgically resected NSCLC specimens. NOS2, COX2, and VEGF immunoreactivity were observed in 48, 48, and 58%, respectively, of the study subjects, and their levels correlated with MVD at the tumor-stromal interphase (P less than or equal to 0.001). More adenocarcinomas and large cell carcinomas displayed overexpression of NOS2 when compared with squamous cell carcinoma (SCC; 0.44; P < 0.001), NOS2 and COX2 levels were found to correlate positively with VEGF status (r = 0.44; P < 0.001, 0.01, and 0.03, respectively). These results attest to the significant interaction of these factors in the angiogenesis of NSCLC. Although neither angiogenic factors nor MVD correlated with patient survival, the latter correlated with tumor clinical stage in both squamous (SCC; 73 BVs/mm(2)) and non-SCC (78 BVs/mm(2)) tumors. These results indicate that angiogenesis is a complex process that involves multiple factors including NOS2, COX2, and VEGF. Furthermore, the role of angiogenesis in the biology of various histological lung cancer types may be different. The complexity of angiogenesis may explain the modest results observed in antiangiogenesis therapy that target a single protein. C1 NCI, Human Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. NCI, Genet Epidemiol Branch, NIH, Bethesda, MD 20892 USA. Armed Forces Inst Pathol, Washington, DC 20306 USA. Mayo Clin, Rochester, MN 55905 USA. RP Harris, CC (reprint author), NCI, Human Carcinogenesis Lab, NIH, Bldg 37,Room 2C05,MSC 4255, Bethesda, MD 20892 USA. NR 43 TC 161 Z9 175 U1 2 U2 2 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD DEC PY 2000 VL 6 IS 12 BP 4739 EP 4744 PG 6 WC Oncology SC Oncology GA 388CQ UT WOS:000166160200022 PM 11156228 ER PT J AU Schleutker, J Matikainen, M Smith, J Koivisto, P Baffoe-Bonnie, A Kainu, T Gillanders, E Sankila, R Pukkala, E Carpten, J Stephan, D Tammela, T Brownstein, M Bailey-Wilson, J Trent, J Kallioniemi, OP AF Schleutker, J Matikainen, M Smith, J Koivisto, P Baffoe-Bonnie, A Kainu, T Gillanders, E Sankila, R Pukkala, E Carpten, J Stephan, D Tammela, T Brownstein, M Bailey-Wilson, J Trent, J Kallioniemi, OP TI A genetic epidemiological study of hereditary prostate cancer (HPC) in Finland: Frequent HPCX linkage in families with late-onset disease SO CLINICAL CANCER RESEARCH LA English DT Article ID SUSCEPTIBILITY LOCUS; CHROMOSOME 1Q; DOMINANT INHERITANCE; RISK; DIAGNOSIS; SUPPORT; HISTORY AB Several predisposition loci for hereditary prostate cancer (HPC) have been suggested, including HPC1 at 1q24-q25 (OMIM #601518) and HPCX at Xq27-q28 (OMIM #300147), Genetically homogeneous populations, such as that of Finland, and distinct subsets of families may help to minimize the genetic heterogeneity that complicates the genetic dissection of complex traits. Here, the role of the HPC1 and HPCX loci in a series of Finnish prostate cancer families was studied, especially in subgroups of families defined by age, number of affected cases, and the mode of disease transmission. DNA samples were collected from 57 Finnish HPC families with at least two living prostate cancer patients. Linkage analysis was carried out with 39 microsatellite markers for the HPC1 region and 22 markers for the HPCX region. The maximum two-point LOD score for the HPCX was 2.05 (marker DXS1205, at 0 = 0.14), whereas HPC1 LOD scores were all negative. In HOMOG3R analyses, significant evidence of heterogeneity was observed. Subgroup analyses performed to explore the nature of this heterogeneity indicated that families with no male-to-male (NMM) transmission and a late age of diagnosis (>65 years) accounted for most of the HPCX-linked cases, The maximum HPCX LOD score in this subgroup was 3.12 (0 = 0.001), Nonparametric sibling pair analyses gave a peak LOD score of 3.04 (P < 0.000093) for the NMM transmission subgroup, No subgroup shelved any positivity for HPC1. This study suggests that the HPCX-linked prostate cancer families represent a distinct subgroup characterized by NMM transmission of disease and late age of diagnosis. C1 Univ Tampere, Inst Med Technol, Canc Genet Lab, FIN-33521 Tampere, Finland. Tampere Univ Hosp, FIN-33521 Tampere, Finland. NHGRI, Canc Genet Branch, NIH, Bethesda, MD 20892 USA. NHGRI, Inherited Dis Res Branch, NIH, Baltimore, MD 21224 USA. Finnish Canc Registry, FIN-00170 Helsinki, Finland. Tampere Univ Hosp, Div Urol, Tampere 33521, Finland. Fox Chase Canc Ctr, Philadelphia, PA 19111 USA. RP Schleutker, J (reprint author), Univ Tampere, Inst Med Technol, Canc Genet Lab, FIN-33521 Tampere, Finland. RI Brownstein, Michael/B-8609-2009; Kallioniemi, Olli/H-5111-2011; Smith, Jeff/C-3484-2012; Kallioniemi, Olli/H-4738-2012; OI Kallioniemi, Olli/0000-0002-3231-0332; Kallioniemi, Olli/0000-0002-3231-0332; Bailey-Wilson, Joan/0000-0002-9153-2920 FU NHGRI NIH HHS [N01-HG-55389] NR 31 TC 66 Z9 66 U1 0 U2 2 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD DEC PY 2000 VL 6 IS 12 BP 4810 EP 4815 PG 6 WC Oncology SC Oncology GA 388CQ UT WOS:000166160200033 PM 11156239 ER PT J AU Dufour, DR Lott, JA Nolte, FS Gretch, DR Koff, RS Seeff, LB AF Dufour, DR Lott, JA Nolte, FS Gretch, DR Koff, RS Seeff, LB TI Diagnosis and monitoring of hepatic injury. I. Performance characteristics of laboratory tests SO CLINICAL CHEMISTRY LA English DT Article; Proceedings Paper CT Annual Meeting of the American-Association-for-Clinical-Chemistry CY JUL 25-26, 1999 CL NEW ORLEANS, LOUISIANA SP Amer Assoc Clin Chem ID INTERNATIONAL NORMALIZED RATIO; B CORE ANTIGEN; GAMMA-GLUTAMYL-TRANSFERASE; RECOMBINANT IMMUNOBLOT ASSAY; POLYMERASE CHAIN-REACTION; C VIRUS-INFECTION; SERUM ALANINE AMINOTRANSFERASE; ALKALINE-PHOSPHATASE ACTIVITY; MITOCHONDRIAL ASPARTATE-AMINOTRANSFERASE; ORAL ANTICOAGULANT-THERAPY AB Purpose: To review information on performance characteristics for tests that are commonly used to identify acute and chronic hepatic injury. Data Sources and Study Selection: A MEDLINE search was performed for key words related to hepatic tests, including quality specifications, aminotransferases, alkaline phosphatase, gamma -glutamyltransferase, bilirubin, albumin, ammonia, and viral markers. Abstracts were reviewed, and articles discussing performance of laboratory tests were selected for review. Additional articles were selected from the references. Guideline Preparation and Review: Drafts of the guidelines were posted on the Internet, presented at the AACC Annual Meeting in 1999, and reviewed by experts. Areas requiring further amplification or literature review were identified for further analysis. Specific recommendations were made based on analysis of published data and evaluated for strength of evidence and clinical impact. The drafts were also reviewed by the Practice Guidelines Committee of the American Association for the Study of Liver Diseases and approved by the committee and the Association's Council. Recommendations: Although many specific recommendations are made in the guidelines, some summary recommendations are discussed here. Alanine aminotransferase is the most important test for recognition of acute and chronic hepatic injury. Performance goals should aim for total error of <10% at the upper reference limit to meet clinical needs in monitoring patients with chronic hepatic injury. Laboratories should have age-adjusted reference limits for enzymes in children, and gender-adjusted reference limits for aminotransferases, -glutamyltransferase, and total bilirubin in adults. The international normalized ratio should not be the sole method for reporting results of prothrombin time in liver disease; additional research is needed to determine the reporting mechanism that best correlates with functional impairment. Harmonization is needed for alanine aminotransferase activity, and improved standardization for hepatitis C viral RNA measurements. (C) 2000 American Association for Clinical Chemistry. C1 Vet Affairs Med Ctr, Pathol & Lab Med Serv, Washington, DC 20422 USA. George Washington Univ, Sch Med, Dept Pathol, Washington, DC 20037 USA. Ohio State Univ, Coll Med, Dept Pathol, Columbus, OH 43210 USA. Emory Univ, Sch Med, Dept Pathol, Atlanta, GA 30322 USA. Emory Univ, Sch Med, Dept Lab Med, Atlanta, GA 30322 USA. Univ Washington, Sch Med, Dept Lab Med, Seattle, WA 98104 USA. Univ Massachusetts, Med Ctr, Dept Med, Worchester, MA USA. NIDDKD, Hepatitis C Programs, NIH, Bethesda, MD 20892 USA. Georgetown Univ, Sch Med, Washington, DC 20037 USA. RP Dufour, DR (reprint author), Vet Adm Med Ctr, Pathol & Lab Med Serv 113, 50 Irving St NW, Washington, DC 20422 USA. NR 263 TC 193 Z9 214 U1 0 U2 14 PU AMER ASSOC CLINICAL CHEMISTRY PI WASHINGTON PA 2101 L STREET NW, SUITE 202, WASHINGTON, DC 20037-1526 USA SN 0009-9147 J9 CLIN CHEM JI Clin. Chem. PD DEC PY 2000 VL 46 IS 12 BP 2027 EP 2049 PG 23 WC Medical Laboratory Technology SC Medical Laboratory Technology GA 380UQ UT WOS:000165722200035 PM 11106349 ER PT J AU Dufour, DR Lott, JA Nolte, FS Gretch, DR Koff, RS Seeff, LB AF Dufour, DR Lott, JA Nolte, FS Gretch, DR Koff, RS Seeff, LB TI Diagnosis and monitoring of hepatic injury. II. Recommendations for use of laboratory tests in screening, diagnosis, and monitoring SO CLINICAL CHEMISTRY LA English DT Article; Proceedings Paper CT Annual Meeting of the American-Association-for-Clinical-Chemistry CY JUL 25-26, 1999 CL NEW ORLEANS, LOUISIANA SP Amer Assoc Clin Chem ID GAMMA-CARBOXY PROTHROMBIN; B-E-ANTIGEN; PRIMARY BILIARY-CIRRHOSIS; PRIMARY SCLEROSING CHOLANGITIS; ALPHA-FETOPROTEIN LEVELS; ALCOHOLIC LIVER-DISEASE; TT-VIRUS-INFECTION; HETEROZYGOUS ALPHA(1)-ANTITRYPSIN DEFICIENCY; ASYMPTOMATIC HEPATOCELLULAR-CARCINOMA; ALANINE AMINOTRANSFERASE LEVELS AB Purpose: To review information on the use of laboratory tests in screening, diagnosis, and monitoring of acute and chronic hepatic injury. Data Sources and Study Selection: A MEDLINE search was performed for key words related to hepatic diseases, including acute hepatitis, chronic hepatitis, alcoholic hepatitis, cirrhosis, hepatocellular carcinoma, and etiologic causes. Abstracts were reviewed, and articles discussing use of laboratory tests selected for review. Additional articles were selected from the references. Guideline Preparation and Review: Drafts of the guidelines were posted on the Internet, presented at the AACC Annual Meeting in 1999, and reviewed by experts. Areas requiring further amplification or literature review were identified for further analysis. Specific recommendations were made based on analysis of published data and evaluated for strength of evidence and clinical impact. Recommendations: Although many specific recommendations are made in the guidelines, only some summary recommendations are listed here. In acute hepatic injury, prothrombin time and, to a lesser extent, total bilirubin are the best indicators of severity of disease. Although ALT is useful for detecting acute and chronic hepatic injury, it is not related to severity of acute hepatic injury and only weakly related to severity of chronic hepatic injury. Specific tests of viral markers should be the initial differential tests in both acute and chronic hepatic injury; when positive, they are also useful for monitoring recovery from hepatitis B and C. (C) 2000 American Association for Clinical Chemistry. C1 Vet Affairs Med Ctr, Pathol & Lab Med Serv, Washington, DC 20422 USA. George Washington Univ, Sch Med, Dept Pathol, Washington, DC 20037 USA. Ohio State Univ, Coll Med, Dept Pathol, Columbus, OH 43210 USA. Emory Univ, Sch Med, Dept Pathol, Atlanta, GA 30322 USA. Emory Univ, Sch Med, Dept Lab Med, Atlanta, GA 30322 USA. Univ Washington, Sch Med, Dept Lab Med, Seattle, WA 98104 USA. Univ Massachusetts, Med Ctr, Dept Med, Worcester, MA 06155 USA. NIDDKD, Hepatitis C Programs, NIH, Bethesda, MD 20892 USA. Georgetown Univ, Sch Med, Washington, DC 20037 USA. RP Dufour, DR (reprint author), Vet Adm Med Ctr, Pathol & Lab Med Serv, 113,50 Irving St SW, Washington, DC 20422 USA. NR 220 TC 147 Z9 162 U1 0 U2 7 PU AMER ASSOC CLINICAL CHEMISTRY PI WASHINGTON PA 2101 L STREET NW, SUITE 202, WASHINGTON, DC 20037-1526 USA SN 0009-9147 J9 CLIN CHEM JI Clin. Chem. PD DEC PY 2000 VL 46 IS 12 BP 2050 EP 2068 PG 19 WC Medical Laboratory Technology SC Medical Laboratory Technology GA 380UQ UT WOS:000165722200036 PM 11106350 ER PT J AU Puck, JM AF Puck, JM TI A disease gene for autosomal hyper-IgM syndrome: More genes associated with more immunodeficiencies SO CLINICAL IMMUNOLOGY LA English DT Editorial Material C1 NHGRI, Genet & Mol Biol Branch, NIH, Bethesda, MD 20892 USA. RP Puck, JM (reprint author), NHGRI, Genet & Mol Biol Branch, NIH, Bethesda, MD 20892 USA. NR 6 TC 3 Z9 3 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1521-6616 J9 CLIN IMMUNOL JI Clin. Immunol. PD DEC PY 2000 VL 97 IS 3 BP 191 EP 192 DI 10.1006/clim.2000.4973 PG 2 WC Immunology SC Immunology GA 383NZ UT WOS:000165890000001 PM 11112357 ER PT J AU Sereti, I Gea-Banacloche, J Kan, MY Hallahan, CW Lane, HC AF Sereti, I Gea-Banacloche, J Kan, MY Hallahan, CW Lane, HC TI Interleukin 2 leads to dose-dependent expression of the alpha chain of the IL-2 receptor on CD25-negative T lymphocytes in the absence of exogenous antigenic stimulation SO CLINICAL IMMUNOLOGY LA English DT Article DE T lymphocytes; cellular activation; cell surface molecules; cytokine receptors; cytokines ID HUMAN-IMMUNODEFICIENCY-VIRUS; HUMAN PERIPHERAL-BLOOD; ACTIVE ANTIRETROVIRAL THERAPY; SUBCUTANEOUS INTERLEUKIN-2; GAMMA-CHAIN; BETA-CHAIN; HIV-INFECTION; B EXPRESS; IN-VITRO; CELLS AB Expression of the alpha chain of the interleukin 2 receptor on T lymphocytes is restricted, increasing in the setting of activation, particularly after antigenic stimulation via the TCR, The effects of IL-2 in vitro on the expression of CD25 and proliferation as well as the cytokine induction in CD25-depleted T cells were studied. CD25-depleted and PBMC of healthy donors were cultured for 7 days with 0, 10, or 100 IU/ml of IL-2. Phenotypic analysis and measurement of cytokines in the culture supernatants were performed. IL-2 led to a dose-dependent induction of the IL-2R alpha chain on both CD4 and CD8 T lymphocytes. In the CD25-depleted cultures, II-a treatment (100 IU/ml) increased the percentage of CD4 T cells expressing CD25 by 30.6% (P = 0.05) and of CD8 T cells by 48.2% (P = 0.01) on day 7 compared, to no treatment. In the PBMC cultures the increase an day 7 was 36.4% for CD4 (P = 0.01) and 50.8% (P = 0.025) for CD8 T lymphocytes. The patterns of cytokine induction in the CD25-depleted and control cultures mere similar with increases of IFN-gamma, GM-CSF, IL-16, TNF alpha; and soluble IL-2 receptor in the IL-2-containing cultures. CFSE experiments demonstrated the proliferative capacity of both CD25-positive and -negative T cells, Interleukin 2 alone can lead to a dose-dependent induction of the alpha chain of its receptor on resting CD4 and CD8 T lymphocytes. IL-2 as a sole stimulant is also associated with generation of a cytokine milieu that includes IFN-gamma GM-CSF, IL-16, and TNF alpha. (C) 2000 Academic Press. C1 NIAID, Clin & Mol Retrovirol Sect, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. NIAID, Immunol Dis Sect, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. RP Lane, HC (reprint author), NIAID, Clin & Mol Retrovirol Sect, Immunoregulat Lab, NIH, Bldg 10,Room 11B-09,10 Ctr Dr MSC 1876, Bethesda, MD 20892 USA. NR 49 TC 20 Z9 20 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1521-6616 J9 CLIN IMMUNOL JI Clin. Immunol. PD DEC PY 2000 VL 97 IS 3 BP 266 EP 276 DI 10.1006/clim.2000.4929 PG 11 WC Immunology SC Immunology GA 383NZ UT WOS:000165890000010 PM 11112366 ER PT J AU Burstein, AH Horton, RL Dunn, T Alfaro, RM Piscitelli, SC Theodore, W AF Burstein, AH Horton, RL Dunn, T Alfaro, RM Piscitelli, SC Theodore, W TI Lack of effect of St John's Wort on carbamazepine pharmacokinetics in healthy volunteers SO CLINICAL PHARMACOLOGY & THERAPEUTICS LA English DT Article ID AUTOINDUCTION; METABOLISM; PLASMA; INDUCTION; EPILEPSY AB Background: St John's Wort is a popular herbal product used by approximately 7% of patients with epilepsy. Previous reports have described reductions in concentrations of CYP3A4 substrates indinavir and cyclosporine (INN, cidosporin) associated with St John's Wort. Objective: Our objective was to determine the effect of St John's Wort on steady state carbamazepine and carbamazepine-10,11-epoxide pharmacokinetics. Methods nad Subjects: Eight healthy volunteers (5 men; age range, 24-43 years) participated in this unblinded study. Subjects received 100 mg of carbamazepine twice daily for 3 days, 200 mg twice daily for 3 days, and then 400 mg once daily for 14 days. Blood samples were collected before and 1, 2, 4, 6, 8, 10, 12, and 24 hours after the dose on day 21. The subjects then took 300 mg of St John's Wort (0.3% hypericin standardized tablet) 3 times daily with meals and with carbamazepine for 14 days. On day 35, blood sampling was repeated. Plasma samples were analyzed for carbamazepine and carbamazepine-10,11-epoxide with HPLC. We compared carbamazepine and carbamazepine-10,11-epoxide noncompartmental pharmacokinetic parameter values before and after St John's Wort with a paired Student t test. Conclusions: The results suggest that treatment with St John's Wart for 14 days did not further induce the clearance of carbamazepine. C1 NINDS, Epilepsy Res Branch, Clin Ctr Nursing Dept, Dept Pharm,NIH, Bethesda, MD 20892 USA. RP Burstein, AH (reprint author), NINDS, Epilepsy Res Branch, Clin Ctr Nursing Dept, Dept Pharm,NIH, Bldg 10,Room 1N257, Bethesda, MD 20892 USA. NR 29 TC 72 Z9 76 U1 4 U2 9 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0009-9236 J9 CLIN PHARMACOL THER JI Clin. Pharmacol. Ther. PD DEC PY 2000 VL 68 IS 6 BP 605 EP 612 DI 10.1067/mcp.2000.111530 PG 8 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 390NM UT WOS:000166302800005 PM 11180020 ER PT J AU Duray, PH AF Duray, PH TI Pathology of melanoma - Preface SO CLINICS IN LABORATORY MEDICINE LA English DT Editorial Material C1 NCI, Div Clin Sci, NIH, Bethesda, MD 20892 USA. RP Duray, PH (reprint author), NCI, Div Clin Sci, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0272-2712 J9 CLIN LAB MED JI Clin. Lab. Med. PD DEC PY 2000 VL 20 IS 4 BP XI EP XII PG 2 WC Medical Laboratory Technology SC Medical Laboratory Technology GA 401VD UT WOS:000166948200001 ER PT J AU Pollock, PM Trent, JM AF Pollock, PM Trent, JM TI The genetics of cutaneous melanoma SO CLINICS IN LABORATORY MEDICINE LA English DT Review ID MULTIPLE PRIMARY MELANOMAS; DYSPLASTIC NEVUS SYNDROME; SPORADIC PRIMARY MELANOMAS; CYCLIN-DEPENDENT KINASES; CLINIC-BASED POPULATION; TUMOR-SUPPRESSOR GENE; FAMILIAL MELANOMA; MALIGNANT-MELANOMA; GERMLINE MUTATIONS; CELL-LINES AB Germline CDKN2A mutations have been identified in approximately 20% of all melanoma families; however, germline mutations occur at a much lower frequency in clinic- or population-based studies, especially in countries where ultraviolet exposure is high. It is likely that additional genes are involved in melanoma susceptibility. It is also likely that additional genes involved in melanoma initiation and progression may be identified through positional cloning studies directed toward chromosomal regions already implicated ill melanoma progression. The advent of new technologies also holds promises for the rapid discovery and characterization of additional genes involved in melanoma pathogenesis. C1 NHGRI, Canc Genet Branch, NIH, Bethesda, MD 20892 USA. RP Pollock, PM (reprint author), NHGRI, Canc Genet Branch, NIH, 9000 Rockville Pike,Bldg 36, Bethesda, MD 20892 USA. NR 119 TC 43 Z9 44 U1 0 U2 1 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0272-2712 J9 CLIN LAB MED JI Clin. Lab. Med. PD DEC PY 2000 VL 20 IS 4 BP 667 EP + PG 25 WC Medical Laboratory Technology SC Medical Laboratory Technology GA 401VD UT WOS:000166948200003 PM 11221509 ER PT J AU Terron, JA AF Terron, JA TI 2-(2-aminoethyl)-quinoline (D-1997): A novel agonist at craniovascular 5-HT1 receptors relevant to migraine therapy SO CNS DRUG REVIEWS LA English DT Review DE D-1997; migraine; quinoline derivatives; serotonin; vascular 5-HT1B/1D receptors; vasoconstriction ID EXTERNAL CAROTID VASOCONSTRICTION; ISOLATED SAPHENOUS-VEIN; VAGOSYMPATHECTOMIZED DOGS; ANTIMIGRAINE DRUGS; PHARMACOLOGICAL EVIDENCE; MEDIATING CONTRACTION; BIOGENIC SUBSTANCES; CANINE; SUMATRIPTAN; 5-HYDROXYTRYPTAMINE AB The functional pharmacological properties of a group of quinoline-derivatives were screened for agonist activity at 5-HT1-like receptors mediating vasoconstriction, In experimental models predictive of antimigraine activity, 2-(2-aminoethyl) quinoline hydrochloride (D-1997) exhibited higher potency and efficacy at vasoconstrictor 5-HT1-like receptors than quipazine. D-1997 was also found to activate a novel vasoconstrictor mechanism in the carotid circulation. It is suggested D-1997 may represent a useful lead in the search for better acute antimigraine therapies. C1 Inst Politecn Nacl, Ctr Invest & Estudios Avanzados, Dept Farmacol & Toxicol, Mexico City, DF, Mexico. RP Terron, JA (reprint author), NIMH, Pharmacol Sect, Bldg 10,Room 2D57,10 Ctr Dr MSC 1514, Bethesda, MD 20892 USA. NR 47 TC 3 Z9 3 U1 0 U2 1 PU NEVA PRESS PI BRANFORD PA P O BOX 347, BRANFORD, CT 06405 USA SN 1080-563X J9 CNS DRUG REV JI CNS Drug Rev. PD WIN PY 2000 VL 6 IS 4 BP 267 EP 277 PG 11 WC Neurosciences; Pharmacology & Pharmacy SC Neurosciences & Neurology; Pharmacology & Pharmacy GA 388CJ UT WOS:000166159600001 ER PT J AU Deng, JP St Clair, M Everett, C Reitman, M Star, RA AF Deng, JP St Clair, M Everett, C Reitman, M Star, RA TI Buprenorphine given after surgery does not alter renal ischemia/reperfusion injury SO COMPARATIVE MEDICINE LA English DT Article ID ANIMALS; RATS; PAIN; TRANSPLANTATION; RECOGNITION; ISCHEMIA; AGONIST; FAILURE; AGENT; MICE AB Background and Purpose: Potential drugs for human acute renal failure are often tested in an animal model of renal ischemia/reperfusion injury. Analgesics are often not given after surgery because of concerns that they would alter renal function. Therefore, we tested whether postoperative analgesia would alter animal health or affect the degree of renal injury. Methods: Mice were subjected to either 32 or 37 minutes of renal ischemia, given two or six doses of buprenorphine or vehicle at 12-hour intervals, and followed for 72 hours. In some animals, we measured body temperature and physical activity by use of telemetry. Results: Animals treated with buprenorphine recovered more rapidly from surgery based on postoperative activity, and had a small but not significant tendency for faster restoration of normal body temperature. Animals treated with buprenorphine had less weight loss after 37 minutes of ischemia, Buprenorphine given after surgery did not influence the degree of renal injury after ischemia/reperfusion. Conclusions: Buprenorphine should be given after renal ischemia-reperfusion surgery because administration of the proper analgesic improved animal health without interfering with the renal ischemia/reperfusion model. Analgesic treatment at the time of the operation and 12 hours after was sufficient. Buprenorphine may reduce the post-surgical stress response, and thus potentially improve the specificity of testing for drugs that reduce or treat renal injury. C1 NIDDKD, Renal Diagnost & Therapeut Unit, NIH, Bethesda, MD 20892 USA. NIDDKD, Lab Anim Sci Sect, NIH, Bethesda, MD 20892 USA. NIDDKD, Diabet Branch, NIH, Bethesda, MD 20892 USA. RP Deng, JP (reprint author), NIDDKD, Renal Diagnost & Therapeut Unit, NIH, 10 Ctr Dr MSC 1268, Bethesda, MD 20892 USA. RI Reitman, Marc/B-4448-2013 OI Reitman, Marc/0000-0002-0426-9475 NR 31 TC 10 Z9 10 U1 0 U2 1 PU AMER ASSOC LABORATORY ANIMAL SCIENCE PI MEMPHIS PA 9190 CRESTWYN HILLS DR, MEMPHIS, TN 38125 USA SN 0023-6764 J9 COMPARATIVE MED JI Comparative Med. PD DEC PY 2000 VL 50 IS 6 BP 628 EP 632 PG 5 WC Veterinary Sciences; Zoology SC Veterinary Sciences; Zoology GA 391PD UT WOS:000166363600012 PM 11200569 ER PT J AU Gohagan, JK AF Gohagan, JK TI The prostate, lung, colorectal and ovarian (PLCO) cancer screening trial - Preface SO CONTROLLED CLINICAL TRIALS LA English DT Editorial Material C1 NCI, Div Canc Prevent, Early Detect Res Grp, Bethesda, MD 20892 USA. RP Gohagan, JK (reprint author), NCI, Div Canc Prevent, Early Detect Res Grp, Bethesda, MD 20892 USA. NR 0 TC 34 Z9 34 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0197-2456 J9 CONTROL CLIN TRIALS JI Controlled Clin. Trials PD DEC PY 2000 VL 21 IS 6 SU S BP 249S EP 250S DI 10.1016/S0197-2456(00)00096-9 PG 2 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 385DT UT WOS:000165987600001 ER PT J AU Gohagan, JK Prorok, PC Hayes, RB Kramer, BS AF Gohagan, JK Prorok, PC Hayes, RB Kramer, BS CA PLCO Project Team TI The Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial of the National Cancer Institute: History, organization, and status SO CONTROLLED CLINICAL TRIALS LA English DT Article DE randomized trial; screening; prostate cancer; colorectal cancer; lung cancer; ovarian cancer; minorities; etiology; biomarkers ID SURVEILLANCE SERIES; INTERPRETING TRENDS; MORTALITY AB The Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial is enrolling 148,000 men and women ages 55-74 at ten screening centers nationwide with balanced randomization to intervention and control arms. For prostate cancer, men receive a digital rectal examination and a blood test for prostate-specific antigen. For lung cancer, men and women receive a posteroanterior view chest X-ray. For colorectal cancer, men and women undergo a 60-cm flexible sigmoidoscopy. For ovarian cancer, women receive a blood test for the CA125 tumor marker and transvaginal ultrasound. Members of the control arm continue with their usual care. Follow-up in both groups will continue for at least 13 years from randomization to assess health status and cause of death. The primary endpoint is mortality from the four PLCO cancers, which accounts for about 53% of all cancer deaths in men and 41% of cancer deaths in women in the United States each year. Blood specimens are collected from screened participants, buccal cell DNA from controls, and histology slides from cases; these are maintained in a biorepository. Participants complete a baseline questionnaire (covering health status and risk factors) and a dietary questionnaire. More than 12,000 participants were enrolled in the pilot phase (concluded in September 1994). Changes in the eligibility criteria followed. As of April 2000, enrollment exceeded 144,500. Data are scanned into designated on-site computers for uploading by participant identification number to the coordinating center for quality checks, archival storage, and preparation of analysis datasets for use by the National Cancer Institute (NCI). Scientific direction is provided by NCI scientists, trial investigators, external consultants, and an independent data safety and monitoring board. Performance and data quality are monitored via data edits, site visits, random record audits, and teleconferences. The PLCO trial is formally endorsed by the American Cancer Society and has been ranked by the American Urological Association as one of the most important prostate cancer studies being conducted. Special efforts to enroll black participants are cosponsored by the U.S. Centers for Disease Control and Prevention. Control Clin Trials 2000;21:251S-272S (C) Elsevier Science Inc. 2000. C1 NCI, Div Canc Prevent, Early Detect Res Grp, EPN 330, Bethesda, MD 20892 USA. NCI, Div Canc Epidemiol, Bethesda, MD 20892 USA. RP Gohagan, JK (reprint author), NCI, Div Canc Prevent, Early Detect Res Grp, EPN 330, 6130 Execut Blvd, Bethesda, MD 20892 USA. NR 27 TC 174 Z9 177 U1 0 U2 4 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0197-2456 J9 CONTROL CLIN TRIALS JI Controlled Clin. Trials PD DEC PY 2000 VL 21 IS 6 SU S BP 251S EP 272S DI 10.1016/S0197-2456(00)00097-0 PG 22 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 385DT UT WOS:000165987600002 PM 11189683 ER PT J AU Prorok, PC Andriole, GL Bresalier, RS Buys, SS Chia, D Crawford, ED Fogel, R Gelmann, EP Gilbert, F Hasson, MA Hayes, RB Johnson, CC Mandel, JS Oberman, A O'Brien, B Oken, MM Rafla, S Reding, D Rutt, W Weissfeld, JL Yokochi, L Gohagan, JK AF Prorok, PC Andriole, GL Bresalier, RS Buys, SS Chia, D Crawford, ED Fogel, R Gelmann, EP Gilbert, F Hasson, MA Hayes, RB Johnson, CC Mandel, JS Oberman, A O'Brien, B Oken, MM Rafla, S Reding, D Rutt, W Weissfeld, JL Yokochi, L Gohagan, JK CA PLCO Project Team TI Design of the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial SO CONTROLLED CLINICAL TRIALS LA English DT Review DE randomized trial; screening; design; prostate cancer; colorectal cancer; lung cancer; ovarian cancer ID FECAL-OCCULT-BLOOD; DIGITAL RECTAL EXAMINATION; SERUM CA-125 LEVELS; RANDOMIZED CONTROLLED TRIAL; TRANSVAGINAL SONOGRAPHY; ULTRASOUND EXAMINATION; RADICAL PROSTATECTOMY; DETECTION-PROJECT; DECISION-ANALYSIS; NATURAL-HISTORY AB The objectives of the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial are to determine in screenees ages 55-74 at entry whether screening with flexible sigmoidoscopy (60-cm sigmoidoscope) can reduce mortality from colorectal cancer, whether screening with chest X-ray can reduce mortality from lung cancer, whether screening men with digital rectal examination (DRE) plus serum prostate-specific antigen (PSA) can reduce mortality from prostate cancer, and whether screening women with CA125 and transvaginal ultrasound (TVU) can reduce mortality from ovarian cancer. Secondary objectives are to assess screening variables other than mortality for each of the interventions including sensitivity, specificity, and positive predictive value; to assess incidence, stage, and survival of cancer cases; and to investigate biologic and/or prognostic characterizations of tumor tissue and biochemical products as intermediate endpoints. The design is a multicenter, two-armed, randomized trial with 37,000 females and 37,000 males in each of the two arms. In the intervention arm, the PSA and CA125 tests are performed at entry, then annually for 5 years. The DRE, TW, and chest X-ray exams are performed at entry and then annually for 3 years. Sigmoidoscopy is performed at entry and then at the 5-year point. Participants in the control arm follow their usual medical care practices. Participants will be followed for at least 13 years from randomization to ascertain all cancers of the prostate, lung, colorectum, and ovary, as well as deaths from all causes. A pilot phase was undertaken to assess the randomization, screening, and data collection procedures of the trial and to estimate design parameters such as compliance and contamination levels. This paper describes eligibility, consent, and other design features of the trial, randomization and screening procedures, and an outline of the follow-up procedures. Sample-size calculations are reported, and a data analysis plan is presented. Control Clin Trials 2000;21:273S-309S (C) Elsevier Science Inc. 2000. C1 NCI, Div Canc Prevent, Early Detect Res Grp, EPN 330, Bethesda, MD 20892 USA. NCI, Div Canc Epidemiol, Bethesda, MD 20892 USA. Washington Univ, Sch Med, St Louis, MO USA. Henry Ford Hlth Syst, Detroit, MI USA. Univ Utah, Hlth Sci Ctr, Salt Lake City, UT USA. Univ Calif Los Angeles, Tissue Typing Lab, Los Angeles, CA USA. Univ Colorado, Hlth Sci Ctr, Denver, CO USA. Georgetown Univ, Washington, DC USA. Pacific Hlth Res Inst, Honolulu, HI USA. Westat Inc, Rockville, MD USA. Univ Minnesota, Minneapolis, MN USA. Univ Alabama, Birmingham, AL USA. Virginia Piper Canc Ctr, Minneapolis, MN USA. Canc Inst Brooklyn, Brooklyn, NY USA. Marshfield Med Res & Educ Fdn, Marshfield, WI USA. Univ Pittsburgh, Inst Canc, Pittsburgh, PA USA. RP Prorok, PC (reprint author), NCI, Div Canc Prevent, Early Detect Res Grp, EPN 330, 6130 Execut Blvd, Bethesda, MD 20892 USA. NR 139 TC 266 Z9 269 U1 1 U2 21 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0197-2456 J9 CONTROL CLIN TRIALS JI Controlled Clin. Trials PD DEC PY 2000 VL 21 IS 6 SU S BP 273S EP 309S DI 10.1016/S0197-2456(00)00098-2 PG 37 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 385DT UT WOS:000165987600003 PM 11189684 ER PT J AU O'Brien, B Nichaman, L Browne, JEH Levin, DL Prorok, PC Gohagan, JK Sullivan, D AF O'Brien, B Nichaman, L Browne, JEH Levin, DL Prorok, PC Gohagan, JK Sullivan, D CA PLCO Project Team TI Coordination and management of a large multicenter screening trial: The Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial SO CONTROLLED CLINICAL TRIALS LA English DT Article DE multicenter clinical trials; data collection; screening; cancer; coordination; operations; management AB The Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial is a large and complex multi-institutional, multifaceted organization with a 20-year horizon. The implementation of the trial began with the creation of an organizational structure that supports strong leadership, cooperation, and effective communication among the trial collaborators including an operational framework for the development, review, and pretest of instruments, data collection, and management procedures; the setting of high-quality standards for training of trial staff; and the development of a comprehensive assessment plan for evaluation of all trial activities. This paper describes the process and methods used in the coordination and management of the PLCO trial. These include the role of the steering committee and its subcommittees and working groups, the establishment of regular and ad hoc communications among collaborators, the training of screening center coordinators and examiners, the PLCO manual of operations and procedures, and the development and implementation of a comprehensive quality assurance plan. Control Clin Trials 2000;21:310S-328S (C) Elsevier Science Inc. 2000. C1 NCI, Div Canc Prevent, Early Detect Res Grp, EPN 330, Bethesda, MD 20892 USA. NCI, Div Canc Prevent, Biometry Res Grp, Bethesda, MD 20892 USA. Westat Inc, Rockville, MD USA. RP Sullivan, D (reprint author), NCI, Div Canc Prevent, Early Detect Res Grp, EPN 330, 6130 Execut Blvd, Bethesda, MD 20892 USA. NR 4 TC 12 Z9 12 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0197-2456 J9 CONTROL CLIN TRIALS JI Controlled Clin. Trials PD DEC PY 2000 VL 21 IS 6 SU S BP 310S EP 328S DI 10.1016/S0197-2456(00)00099-4 PG 19 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 385DT UT WOS:000165987600004 PM 11189685 ER PT J AU Hasson, MA Fagerstrom, RM Kahane, DC Walsh, JH Myers, MH Caughman, C Wenzel, B Haralson, JC Flickinger, LM Turner, LM AF Hasson, MA Fagerstrom, RM Kahane, DC Walsh, JH Myers, MH Caughman, C Wenzel, B Haralson, JC Flickinger, LM Turner, LM CA PLCO Project Team TI Design and evolution of the data management systems in the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial SO CONTROLLED CLINICAL TRIALS LA English DT Article DE software design; database management systems; automated data processing; data collection AB This paper describes the design and evolution of the data management systems developed in support of the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial. These systems span platforms from stand-alone computers to distributed systems on local area net-works to mainframes. Allowing all of these systems to share appropriate information electronically introduces integration, synchronization, testing, and support challenges. For each platform, applications were developed to handle data entry, editing, trial management, reporting, telecommunications, and data sharing. Approaches to issues such as level of data access, integration with other, existing applications, and handling the expansion of the protocol are discussed. Control Clin Trials 2000;21:329S-348S (C) Elsevier Science Inc. 2000. C1 NCI, Div Canc Prevent, Biometry Res Grp, Bethesda, MD 20892 USA. Westat Inc, Rockville, MD USA. NOVA Res Co, Bethesda, MD USA. Marshfield Med Res & Educ Fdn, Marshfield, WI USA. Henry Ford Hlth Syst, Detroit, MI USA. Pacific Hlth Res Inst, Honolulu, HI USA. RP Hasson, MA (reprint author), NCI, Div Canc Prevent, Early Detect Res Grp, EPN 330, 6130 Execut Blvd, Bethesda, MD 20892 USA. NR 34 TC 21 Z9 22 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0197-2456 J9 CONTROL CLIN TRIALS JI Controlled Clin. Trials PD DEC PY 2000 VL 21 IS 6 SU S BP 329S EP 348S DI 10.1016/S0197-2456(00)00100-8 PG 20 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 385DT UT WOS:000165987600005 PM 11189686 ER PT J AU Hayes, RB Reding, D Kopp, W Subar, AF Bhat, N Rothman, N Caporaso, N Ziegler, RG Johnson, CC Weissfeld, JL Hoover, RN Hartge, P Palace, C Gohagan, JK AF Hayes, RB Reding, D Kopp, W Subar, AF Bhat, N Rothman, N Caporaso, N Ziegler, RG Johnson, CC Weissfeld, JL Hoover, RN Hartge, P Palace, C Gohagan, JK CA PLCO Project Team TI Etiologic and early marker studies in the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial SO CONTROLLED CLINICAL TRIALS LA English DT Article DE etiology; genetic markers; health questionnaire; risk factors; dietary questionnaire AB The Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial, which is randomizing 74,000 screening arm participants (37,000 men, 37,000 women; ages 55-74) and an equal number of nonscreened controls, is a unique setting for the investigation of the etiology of cancer and other diseases and for the evaluation of potential molecular markers of early disease. At entry, baseline information is collected by questionnaire on dietary intake, tobacco and alcohol use, reproductive history (for women), family history of cancer, use of selected drugs, and other selected risk factors. Blood samples collected at the baseline screening exam are aliquoted to serum, plasma, red blood cell, and buffy coat fractions. At the next two annual screening visits, serum samples are collected. At the third annual reexamination, cryopreserved whole blood is obtained, in addition to serum, plasma, red blood cell, and buffy coat fractions. At the fourth and fifth years, serum, plasma, and buffy coat are collected. All blood samples are shipped to a central repository for long-term storage at -70 degreesC. Dietary questionnaires and buccal cells for DNA analysis are obtained from nonscreened controls. Cancer cases are identified through annual follow-up questionnaires, and all deaths are identified through vital status tracing mechanisms. Procedures are being developed to obtain archival pathologic material for selected cases of cancer and related diseases. Initial investigations are focusing on the etiology of colorectal cancer and on the operative characteristics of tests for the early detection of colorectal and prostate cancer. Control Clin Trials 2000;21:349S-355S (C) Elsevier Science Inc. 2000. C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Marshfield Med Res & Educ Fdn, Marshfield, WI USA. Sci Applicat Int Corp, Frederick, MD USA. NCI, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. Henry Ford Hlth Syst, Detroit, MI USA. Univ Pittsburgh, Inst Canc, Pittsburgh, PA USA. Westat Inc, Rockville, MD USA. NCI, Div Canc Prevent, Early Detect Res Grp, EPN 330, Bethesda, MD 20892 USA. RP Hayes, RB (reprint author), NCI, Div Canc Prevent, Early Detect Res Grp, EPN 330, 6130 Execut Blvd, Bethesda, MD 20892 USA. NR 6 TC 60 Z9 60 U1 0 U2 4 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0197-2456 J9 CONTROL CLIN TRIALS JI Controlled Clin. Trials PD DEC PY 2000 VL 21 IS 6 SU S BP 349S EP 355S DI 10.1016/S0197-2456(00)00101-X PG 7 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 385DT UT WOS:000165987600006 PM 11189687 ER PT J AU Simpson, NK Johnson, CC Ogden, SL Gamito, E Trocky, N McGuire, C Martin, J Barrow, S Lamerato, L Flickinger, LM Broski, KG Engelhard, D Hilke, C Bonk, J Gahagan, B Gren, LH Childs, J Lappe, K Fouad, M Thompson, J Sullivan, D AF Simpson, NK Johnson, CC Ogden, SL Gamito, E Trocky, N McGuire, C Martin, J Barrow, S Lamerato, L Flickinger, LM Broski, KG Engelhard, D Hilke, C Bonk, J Gahagan, B Gren, LH Childs, J Lappe, K Fouad, M Thompson, J Sullivan, D CA PLCO Project Team TI Recruitment strategies in the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial: The first six years SO CONTROLLED CLINICAL TRIALS LA English DT Article DE recruitment; patient selection; cancer screening; randomized clinical trial ID HEALTH; PARTICIPATION AB The Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial has a total enrollment goal of almost 150,000 participants. These participants are being recruited at ten screening centers across the United States. All screening centers tested recruitment methodologies during a 1-year pilot phase. The main phase of recruitment was planned to take place over a 3-year period. The majority of participants are being recruited during the main phase of the study. Each of the screening centers tailors recruitment to its individual catchment area. Recruitment strategies in the PLCO trial are described. As the trial began, several protocol changes were made to help to increase enrollment. The National Cancer Institute (NCI) initiated recruitment efforts at the national level. The individual screening centers describe some of the specific recruitment experiences encountered. As the study progressed, the NCI implemented special initiatives to increase the enrollment of minority participants. Control Clin Trials 2000; 21:356S-378S (C) Elsevier Science Inc. 2000. C1 NCI, Div Canc Prevent, Early Detect Res Grp, EPN 330, Bethesda, MD 20892 USA. Henry Ford Hlth Syst, Detroit, MI USA. Univ Colorado, Hlth Sci Ctr, Denver, CO USA. Georgetown Univ, Washington, DC USA. Pacific Hlth Res Inst, Honolulu, HI USA. Univ Minnesota, Minneapolis, MN USA. Washington Univ, Sch Med, St Louis, MO USA. Univ Pittsburgh, Inst Canc, Pittsburgh, PA USA. Univ Utah, Hlth Sci Ctr, Salt Lake City, UT USA. Marshfield Med Res & Educ Fdn, Marshfield, WI USA. Univ Alabama, Birmingham, AL USA. RP Sullivan, D (reprint author), NCI, Div Canc Prevent, Early Detect Res Grp, EPN 330, 6130 Execut Blvd, Bethesda, MD 20892 USA. EM ds255j@nih.gov NR 9 TC 34 Z9 35 U1 0 U2 2 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0197-2456 J9 CONTROL CLIN TRIALS JI Controlled Clin. Trials PD DEC PY 2000 VL 21 IS 6 SU S BP 356S EP 378S DI 10.1016/S0197-2456(00)00102-1 PG 23 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 385DT UT WOS:000165987600007 PM 11189688 ER PT J AU Stallings, FL Ford, ME Simpson, NK Fouad, M Jernigan, JC Trauth, JM Miller, DS AF Stallings, FL Ford, ME Simpson, NK Fouad, M Jernigan, JC Trauth, JM Miller, DS CA PLCO Project Team TI Black participation in the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial SO CONTROLLED CLINICAL TRIALS LA English DT Article DE screening; cancer; patient selection; minority groups ID CLINICAL-TRIALS; AFRICAN-AMERICANS; HEALTH-CARE; PREVENTION; WOMEN; SURVIVAL; RACE; REPRESENTATION; RECRUITMENT; ENROLLMENT AB The primary goal of the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial is to learn whether widespread use of screening tests to detect these cancers will reduce associated mortality. Blacks have the highest age-adjusted cancer incidence and mortality rates of any population group in the United States, but several barriers to their participation in clinical research such as the PLCO trial exist. These barriers involve sociocultural, economic, and individual factors, as well as factors inherent in trial designs. Population diversity in the PLCO trial is necessary to preserve scientific validity and generalizability of trial results. Therefore, the National Cancer Institute and the Centers for Disease Control and Prevention are collaborating to ensure adequate representation of blacks in the PLCO trial. For example, the agencies have funded several new activities designed to better understand and overcome barriers to participation in the trial. These activities include the African American Men Project, a randomized trial designed to evaluate the efficacy of three-increasingly intensive recruitment interventions in recruiting black men; the establishment of a minority-focused PLCO trial screening center, a study to identify factors that influenced the decisions of black women recruited to participate in the PLCO trial; and a study to examine the psychosocial factors that influence blacks' decision making to engage in cancer screening and participation in research similar to the PLCO trial. The results of these activities will allow for a more thorough examination of cancer-related issues of importance to blacks and will help shed light on factors that influence their decisions to participate in cancer screening and prevention clinical trials. Control Clin Trials 2000;21:379S-389S (C) Elsevier Science Inc. 2000. C1 NCI, Div Canc Prevent, Early Detect Res Grp, EPN 330, Bethesda, MD 20892 USA. Ctr Dis Control & Prevent, Div Canc Prevent & Control, Natl Ctr Chron Dis Prevent & Hlth Promot, Atlanta, GA USA. Henry Ford Hlth Syst, Ctr Med Treatment Effectiveness Programs Diverse, Detroit, MI USA. Univ Alabama, Birmingham, AL USA. Univ Pittsburgh, Grad Sch Publ Hlth, Pittsburgh, PA USA. RP Stallings, FL (reprint author), NCI, Div Canc Prevent, Early Detect Res Grp, EPN 330, 6130 Execut Blvd, Bethesda, MD 20892 USA. NR 45 TC 36 Z9 36 U1 2 U2 3 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0197-2456 J9 CONTROL CLIN TRIALS JI Controlled Clin. Trials PD DEC PY 2000 VL 21 IS 6 SU S BP 379S EP 389S DI 10.1016/S0197-2456(00)00093-3 PG 11 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 385DT UT WOS:000165987600008 PM 11189689 ER PT J AU Weissfeld, JL Fagerstrom, RM O'Brien, B AF Weissfeld, JL Fagerstrom, RM O'Brien, B CA PLCO Project Team TI Quality control of cancer screening examination procedures in the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial SO CONTROLLED CLINICAL TRIALS LA English DT Article DE quality control; cancer screening; procedures; data analysis ID TRANSVAGINAL SONOGRAPHY; POSTMENOPAUSAL WOMEN; VOLUME; HOSPITALS AB Investigators for the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial describe quality control procedures for the digital rectal examination, ovarian palpation examination, transvaginal ultrasound, chest X-ray, and flexible sigmoidoscopy. These cancer screening tests are subjective and difficult to standardize. PLCO quality control procedures aim to measure and, where possible, reduce variation, across examiner and screening center, with respect to cancer screening test performance. Initial protocols stressed examiner qualifications, experience, and training; equipment specifications; examination procedures; and definitions for positive tests. The PLCO quality assurance subcommittee developed a final quality assurance plan, which included central approval and registration of PLCO examiners, direct observation of screening test performance during periodic site visits by the National Cancer Institute and coordinating center auditors, periodic analysis of screening test data, and procedures for independently duplicating or reviewing selected examinations. For each modality, the periodic data analyses examine the test-positive and the test-inadequate proportions and aim to identify divergent centers or examiners. Procedures for duplicating examinations specify feasible sample sizes for precise estimates of agreement between examiners, at each center, for each screening test modality, and over a 1-year period. These quality control procedures will help characterize the consistency and reliability of the PLCO cancer screening tests. Control Clin Trials 2000;21:390S-399S (C) Elsevier Science Inc. 2000. C1 NCI, Div Canc Prevent, Biometry Res Grp, Bethesda, MD 20892 USA. Univ Pittsburgh, Inst Canc, Pittsburgh, PA USA. Westat Inc, Rockville, MD USA. RP Weissfeld, JL (reprint author), NCI, Div Canc Prevent, Early Detect Res Grp, EPN 330, 6130 Execut Blvd, Bethesda, MD 20892 USA. NR 19 TC 10 Z9 10 U1 0 U2 5 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0197-2456 J9 CONTROL CLIN TRIALS JI Controlled Clin. Trials PD DEC PY 2000 VL 21 IS 6 SU S BP 390S EP 399S DI 10.1016/S0197-2456(00)00094-5 PG 10 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 385DT UT WOS:000165987600009 PM 11189690 ER PT J AU Miller, AB Yurgalevitch, S Weissfeld, JL AF Miller, AB Yurgalevitch, S Weissfeld, JL CA PLCO Project Team TI Death review process in the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial SO CONTROLLED CLINICAL TRIALS LA English DT Article DE screening trial; death review ID MORTALITY AB The procedures that have been adopted in the Prostate, Lung, Colorectal and Ovarian (PLCO) Cancer Screening Trial are described. These procedures have been designed to ensure unbiased decisions on the underlying cause of death for all confirmed or suspected PLCO cancers. A death review committee with a nonvoting chair and three experienced reviewers as members has been appointed. After an initial pilot study, the procedures have been instituted and are working well. Control Clin Trials 2000;21:400S-406S (C) Elsevier Science Inc. 2000. C1 Deutsch Krebsforschungszentrum, Div Clin Epidemiol, D-6900 Heidelberg, Germany. Westat Inc, Rockville, MD USA. Univ Pittsburgh, Inst Canc, Pittsburgh, PA USA. RP Miller, AB (reprint author), NCI, Div Canc Prevent, Early Detect Res Grp, EPN 330, 6130 Execut Blvd, Bethesda, MD 20892 USA. NR 5 TC 40 Z9 40 U1 1 U2 3 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0197-2456 J9 CONTROL CLIN TRIALS JI Controlled Clin. Trials PD DEC PY 2000 VL 21 IS 6 SU S BP 400S EP 406S DI 10.1016/S0197-2456(00)00095-7 PG 7 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 385DT UT WOS:000165987600010 PM 11189691 ER PT J AU Proschan, MA Waclawiw, MA AF Proschan, MA Waclawiw, MA TI Practical guidelines for multiplicity adjustment in clinical trials SO CONTROLLED CLINICAL TRIALS LA English DT Article DE multiple comparisons; familywise error rate ID PREVENTION; DISEASE AB Multiplicity in clinical trials may appear under several different guises: multiple endpoints, multiple treatment arm comparisons, and multiple looks at the data during interim monitoring, to name a few. It is well recognized by statisticians and nonstatisticians alike that multiplicity inflates the type I error rate of the experiment, and this has prompted the development of many multiple comparison adjustment procedures. What has remained one of the thornier and more controversial points of contention among trialists today is the philosophy surrounding the need for multiplicity adjustment in clinical trials. This paper provides guidelines on how to deal with this complex issue in a practical manner. Through a series of scenarios and examples, we illustrate the fundamental issues surrounding the concept of multiplicity and point to some key questions one should ask when deliberating on the necessity and extent of adjustment for multiple comparisons. (C) Elsevier Science Inc. 2000. C1 NHLBI, Off Biostat Res, Bethesda, MD 20892 USA. RP Proschan, MA (reprint author), Rockledge Ctr 2, 6701 Rockledge Dr,Room 8222,MSC 7938, Bethesda, MD 20892 USA. NR 21 TC 95 Z9 96 U1 0 U2 4 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0197-2456 J9 CONTROL CLIN TRIALS JI Controlled Clin. Trials PD DEC PY 2000 VL 21 IS 6 BP 527 EP 539 DI 10.1016/S0197-2456(00)00106-9 PG 13 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 384KF UT WOS:000165940800001 PM 11146147 ER PT J AU Alexander, E Susla, GM Burstein, AH Brown, D Ognibene, FP AF Alexander, E Susla, GM Burstein, AH Brown, D Ognibene, FP TI Retrospective evaluation of commonly used equations to predict energy expenditure in mechanically ventilated critically ill patients SO CRITICAL CARE MEDICINE LA English DT Meeting Abstract C1 Univ Tennessee, Reg Med Ctr, Memphis, TN USA. NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0090-3493 J9 CRIT CARE MED JI Crit. Care Med. PD DEC PY 2000 VL 28 IS 12 SU S MA 272 BP A105 EP A105 PG 1 WC Critical Care Medicine SC General & Internal Medicine GA 388ZD UT WOS:000166212200273 ER PT J AU Cui, XZ Parent, C Banks, S Gerstenberger, E Fitz, Y Danner, RL Natanson, C Eichacker, PQ AF Cui, XZ Parent, C Banks, S Gerstenberger, E Fitz, Y Danner, RL Natanson, C Eichacker, PQ TI Compared to its use alone, TNF soluble receptor (TNFR : Fc) with fluid therapy reduces survival in septic rats SO CRITICAL CARE MEDICINE LA English DT Meeting Abstract C1 NIH, Bethesda, MD 20892 USA. NR 0 TC 1 Z9 1 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0090-3493 J9 CRIT CARE MED JI Crit. Care Med. PD DEC PY 2000 VL 28 IS 12 SU S MA 372 BP A132 EP A132 PG 1 WC Critical Care Medicine SC General & Internal Medicine GA 388ZD UT WOS:000166212200372 ER PT J AU Ezzeddine, MA Oliveira, J Sheik, S Flibotte, J Greer, D Ougorets, I Rosand, J Ogilvy, C Carter, B Rordorf, G Koroshetz, WJ McDonald, CT AF Ezzeddine, MA Oliveira, J Sheik, S Flibotte, J Greer, D Ougorets, I Rosand, J Ogilvy, C Carter, B Rordorf, G Koroshetz, WJ McDonald, CT TI Serum amylase correlates with the clinical severity of subarachnoid hemorrhage SO CRITICAL CARE MEDICINE LA English DT Meeting Abstract C1 NIH, Bethesda, MD 20892 USA. Massachusetts Gen Hosp, Boston, MA 02114 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0090-3493 J9 CRIT CARE MED JI Crit. Care Med. PD DEC PY 2000 VL 28 IS 12 SU S MA 647 BP A207 EP A207 PG 1 WC Critical Care Medicine SC General & Internal Medicine GA 388ZD UT WOS:000166212200647 ER PT J AU Ezzeddine, MA Oleiveira, J Flibotte, J Greer, D Ougoretz, I Rosand, J Rordorf, G Schwamm, L Buonnano, F Koroshetz, WJ Mcdonald, CT AF Ezzeddine, MA Oleiveira, J Flibotte, J Greer, D Ougoretz, I Rosand, J Rordorf, G Schwamm, L Buonnano, F Koroshetz, WJ Mcdonald, CT TI Predictors of extubation failure of mechanically ventilated patients in the neuroscience intensive care unit SO CRITICAL CARE MEDICINE LA English DT Meeting Abstract C1 NIH, Bethesda, MD 20892 USA. Massachusetts Gen Hosp, Boston, MA 02114 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0090-3493 J9 CRIT CARE MED JI Crit. Care Med. PD DEC PY 2000 VL 28 IS 12 SU S MA 251 BP A99 EP A99 PG 1 WC Critical Care Medicine SC General & Internal Medicine GA 388ZD UT WOS:000166212200252 ER PT J AU Nanavaty, UB Gladwin, MT Cowan, MJ Pawliczak, R Logun, C Doniger, J Shelhamer, JH AF Nanavaty, UB Gladwin, MT Cowan, MJ Pawliczak, R Logun, C Doniger, J Shelhamer, JH TI Investigations into pathways leading to oxidative stress induced cell death in lung epithelial cells. SO CRITICAL CARE MEDICINE LA English DT Meeting Abstract C1 Ctr Clin, Bethesda, MD USA. NR 0 TC 0 Z9 0 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0090-3493 J9 CRIT CARE MED JI Crit. Care Med. PD DEC PY 2000 VL 28 IS 12 SU S MA 4 BP A27 EP A27 PG 1 WC Critical Care Medicine SC General & Internal Medicine GA 388ZD UT WOS:000166212200005 ER PT J AU Illei, GG Lipsky, PE AF Illei, GG Lipsky, PE TI Novel, non-antigen-specific therapeutic approaches to autoimmune/inflammatory diseases SO CURRENT OPINION IN IMMUNOLOGY LA English DT Review ID NECROSIS-FACTOR-ALPHA; CHIMERIC MONOCLONAL-ANTIBODY; INTERLEUKIN-1 RECEPTOR ANTAGONIST; RHEUMATOID-ARTHRITIS; CROHNS-DISEASE; FUSION PROTEIN; TRIAL; CA2; INFLIXIMAB; METHOTREXATE AB Anti-TNF therapy has made a major impact on the treatment of inflammatory arthritides and Crohn's disease. It leads to prompt and prolonged clinical response, even in patients refractory to conventional therapy. Moreover, the progression of joint damage noted in patients with rheumatoid arthritis treated with methotrexate was prevented by an anti-TNF-alpha antibody, suggesting a genuine disease-modifying potential of TNF-alpha blockade. C1 NIAMSD, NIH, Bethesda, MD 20892 USA. RP Illei, GG (reprint author), NIAMSD, NIH, 9000 Rockville Pike,Bldg 10, Bethesda, MD 20892 USA. NR 40 TC 38 Z9 38 U1 0 U2 0 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0952-7915 J9 CURR OPIN IMMUNOL JI Curr. Opin. Immunol. PD DEC PY 2000 VL 12 IS 6 BP 712 EP 718 DI 10.1016/S0952-7915(00)00167-9 PG 7 WC Immunology SC Immunology GA 372DA UT WOS:000165218000016 PM 11102777 ER PT J AU Winters, TA AF Winters, TA TI Gene targeted agents: New opportunities for rational drug development SO CURRENT OPINION IN MOLECULAR THERAPEUTICS LA English DT Review DE DNA damage; DNA repair; gene therapy; recombination; transcription; TFO; triple-helix ID TRIPLE-HELIX FORMATION; PEPTIDE NUCLEIC-ACID; CHIMERIC RNA/DNA OLIGONUCLEOTIDES; HUMAN RHODOPSIN GENE; PHOTO-CROSS-LINKING; FORMING OLIGONUCLEOTIDES; MAMMALIAN-CELLS; TRANSCRIPTION ELONGATION; IN-VITRO; MINOR-GROOVE AB In addition to traditional drug development methods designed to modulate the activity of protein targets, knowledge of disease gene DNA sequences provides an opportunity for the highly rational design of therapeutic agents that act at the DNA level through sequence-specific interactions. Among the ligands capable of binding DNA in a precise, sequence-specific manner are oligonucleotides, peptide nucleic acids and polyamides. Various strategies employing these agents to either transiently or permanently alter gene expression have been investigated over the past decade. During the past two to three years, important steps have been taken to illustrate the therapeutic potential of these ligands. Triple-helix (triplex) forming oligonucleotides have been particularly effective DNA-targeting agents with a wide range of applications, including the positive and negative transcriptional regulation of target genes, as well as the controlled delivery of site-specific mutations. This review will focus upon recent advances involving the use of sequence-specific DNA-binding ligands to modify gene expression and/or structure, with particular emphasis on the use of triplex-forming oligonucleotides in these roles. C1 NIH, Dept Nucl Med, Bethesda, MD 20892 USA. RP Winters, TA (reprint author), NIH, Dept Nucl Med, Bethesda, MD 20892 USA. NR 127 TC 25 Z9 27 U1 0 U2 4 PU PHARMAPRESS LTD PI LONDON PA MIDDLESEX HOUSE, 34-42 CLEVELAND ST, LONDON W1P 6LB, ENGLAND SN 1464-8431 J9 CURR OPIN MOL THER JI Curr. Opin. Mol. Ther. PD DEC PY 2000 VL 2 IS 6 BP 670 EP 681 PG 12 WC Biotechnology & Applied Microbiology; Medicine, Research & Experimental SC Biotechnology & Applied Microbiology; Research & Experimental Medicine GA 388BH UT WOS:000166157200008 PM 11249745 ER PT J AU Hurley, JH Tsujishita, Y Pearson, MA AF Hurley, JH Tsujishita, Y Pearson, MA TI Floundering about at cell membranes: a structural view of phospholipid signaling SO CURRENT OPINION IN STRUCTURAL BIOLOGY LA English DT Article ID PHOSPHATIDYLETHANOLAMINE-BINDING PROTEIN; PHOSPHATIDYLINOSITOL PHOSPHATE KINASE; CRYSTAL-STRUCTURE; FYVE DOMAIN; SUGGESTS; FAMILY; A(2) AB Structures are now available for the majority of the enzyme families involved in the phosphorylation, dephosphorylation and hydrolysis of signaling phospholipids. Lipid kinase and phosphatase structures recapitulate catalytic motifs involved in protein phosphorylation and dephosphorylation, whereas cytosolic phospholipase Ap manifests novel catalytic geometry. Structures have been determined for most known intracellular phospholipid 'receptor' domains, both those that bind membrane-embedded phospholipids and those that bind lipid monomers. C1 NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Hurley, JH (reprint author), NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. NR 31 TC 25 Z9 25 U1 0 U2 3 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0959-440X J9 CURR OPIN STRUC BIOL JI Curr. Opin. Struct. Biol. PD DEC PY 2000 VL 10 IS 6 BP 737 EP 743 PG 7 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 394DQ UT WOS:000166509100016 PM 11114512 ER PT J AU Dorman, SE Holland, SM AF Dorman, SE Holland, SM TI Interferon-gamma and interleukin-12 pathway defects and human disease SO CYTOKINE & GROWTH FACTOR REVIEWS LA English DT Review DE interferon-gamma; interferon-gamma receptor deficiency; interleukin-12; tuberculosis; mycobacteria ID BACILLE CALMETTE-GUERIN; CELL STIMULATORY FACTOR; MYCOBACTERIUM-AVIUM INFECTION; VIRUS TYPE-1 INFECTION; CD4+ T-CELLS; IFN-GAMMA; RECEPTOR DEFICIENCY; TYROSINE PHOSPHORYLATION; MURINE MACROPHAGES; LEISHMANIA-MAJOR AB A genetic component to human mycobacterial disease susceptibility has long been postulated. Over the past five years, mutations in the interferon-gamma (IFN gamma) receptor, IL-12 receptor beta 1 (IL-12R beta 1), and IL-12 p40 genes have been recognized. These mutations are associated with heightened susceptibility to disease caused by intracellular pathogens including nontuberculous mycobacteria, vaccine-associated bacille Calmette Guerin (BCG), Salmonella species, and some viruses. We describe the genotype-phenotype correlations in IFN gamma receptor, IL-12R beta 1, and IL-12 p40 deficiency, and discuss how study of these diseases has enhanced knowledge of human host defense against mycobacteria and other intracellular pathogens. Published by Elsevier Science Ltd. C1 NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. RP Holland, SM (reprint author), NIAID, Host Def Lab, NIH, Bldg 10,Room 11N103,10 Ctr Dr,MSC 1886, Bethesda, MD 20892 USA. NR 95 TC 212 Z9 224 U1 0 U2 7 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 1359-6101 J9 CYTOKINE GROWTH F R JI Cytokine Growth Factor Rev. PD DEC PY 2000 VL 11 IS 4 BP 321 EP 333 DI 10.1016/S1359-6101(00)00010-1 PG 13 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 357AL UT WOS:000089476300005 PM 10959079 ER PT J AU Yamada, M Murata, T Hirose, S Lavorgna, G Suzuki, E Ueda, H AF Yamada, M Murata, T Hirose, S Lavorgna, G Suzuki, E Ueda, H TI Temporally restricted expression of transcription factor beta FTZ-F1: significance for embryogenesis, molting and metamorphosis in Drosophila melanogaster SO DEVELOPMENT LA English DT Article DE FTZ-F1; nuclear hormone receptor; Drosophila melanogaster; ecdysone; molting; metamorphosis; BR-C ID STEROID-HORMONE ECDYSONE; HOMEODOMAIN PROTEIN FTZ; REGULATED GENE DISRUPT; NUCLEAR RECEPTOR; PUPAL CUTICLE; BROAD-COMPLEX; SALIVARY-GLANDS; BINDING PROTEIN; FUSHI-TARAZU; BOMBYX-MORI AB FTZ-F1, a member of the nuclear receptor superfamily, has been implicated in the activation of the segmentation gene fushi tarazu during early embryogenesis of Drosophila melanogaster, We found that an isoform of FTZ-F1, beta FTZ-F1, is expressed in the nuclei of almost all tissues slightly before the first and second larval ecdysis and before pupation, Severely affected ftz-f1 mutants display an embryonic lethal phenotype, but can be rescued by ectopic expression of beta FTZ-F1 during the period of endogenous beta FTZ-F1 expression in the wild type. The resulting larvae are not able to molt, but this activity is rescued again by forced expression of beta FTZ-F1, allowing progression to the next larval instar stage. On the other hand, premature expression of beta FTZ-F1 in wild-type larvae at mid-first instar or mid-second instar stages causes defects in the molting process. Sensitive periods were found to be around the time of peak ecdysteroid levels and slightly before the start of endogenous beta FTZ-F1 expression. A hypomorphic ftz-f1 mutant that arrests in the prepupal stage can also be rescued by ectopic, time-specific expression of beta FTZ-F1, Failure of salivary gland histolysis, one of the phenotypes of the ftz-f1 mutant, is rescued by forced expression of the ftz-f1 downstream gene BR-C during the late prepupal period, These results suggest that beta FTZ-F1 regulates genes associated with ecdysis and metamorphosis, and that the exact timing of its action in the ecdysone-induced gene cascade is important for proper development. C1 Grad Univ Adv Studies, Dept Genet, Mishima, Shizuoka 4118540, Japan. Natl Inst Genet, Dept Dev Genet, Mishima, Shizuoka 4118540, Japan. NCI, Mol Cell Biol Lab, NIH, Bethesda, MD 20892 USA. Univ Tokyo, Inst Med Sci, Dept Fine Morphol, Minato Ku, Tokyo 1088639, Japan. Japan Sci & Technol Corp, CREST, Osaka, Japan. RP Ueda, H (reprint author), Grad Univ Adv Studies, Dept Genet, Mishima, Shizuoka 4118540, Japan. RI UEDA, Hitoshi/B-1455-2011 NR 43 TC 79 Z9 82 U1 1 U2 13 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0950-1991 J9 DEVELOPMENT JI Development PD DEC PY 2000 VL 127 IS 23 BP 5083 EP 5092 PG 10 WC Developmental Biology SC Developmental Biology GA 384UA UT WOS:000165962900007 PM 11060234 ER PT J AU Clifford, R Lee, MH Nayak, S Ohmachi, M Giorgini, F Schedl, T AF Clifford, R Lee, MH Nayak, S Ohmachi, M Giorgini, F Schedl, T TI FOG-2, a novel F-box containing protein, associates with the GLD-1 RNA binding protein and directs male sex determination in the C-elegans hermaphrodite germline SO DEVELOPMENT LA English DT Article DE translational repression; GLD-1; FOG-2; F-box; germline sex determination; tra-2 mRNA; Caenorhabditis elegans ID MULTIPLE SEQUENCE ALIGNMENT; 3' UNTRANSLATED REGION; DETERMINING GENE TRA-2; OF-FUNCTION MUTATIONS; SPERM-OOCYTE SWITCH; CAENORHABDITIS-ELEGANS; SIGNAL-TRANSDUCTION; 2-HYBRID SYSTEM; KH DOMAIN; LINE AB Male sex determination in the Caenorhabditis elegans hermaphrodite germline requires translational repression of tra-2 mRNA by the GLD-1 RNA binding protein. We cloned fog-2 by finding that its gene product physically interacts with GLD-1, forming a FOG-2/GLD-1/tra-2 3'untranslated region ternary complex. FOG-2 has an N-terminal F-box and a novel C-terminal domain called FTH, Canonical F-box proteins act as bridging components of the SCF ubiquitin ligase complex; the N-terminal F-box binds a Skp1 homolog, recruiting ubiquination machinery, while a C-terminal protein-protein interaction domain binds a specific substrate for degradation. However, since both fog-2 and gld-1 are necessary for spermatogenesis, FOG-2 cannot target GLD-1 for ubiquitin-mediated degradation. We propose that FOG-2 also acts as a bridge, bringing GLD-1 bound to tra-2 mRNA into a multiprotein translational repression complex, thus representing a novel function for an F-box protein. fog-2 is a member of a large, apparently rapidly evolving, C. elegans gene family that has expanded, in part, by local duplications; fog-2 related genes have not been found outside nematodes. fog-2 may have arisen during evolution of self-fertile hermaphroditism from an ancestral female/male species. C1 Washington Univ, Sch Med, Dept Genet, St Louis, MO 63110 USA. NCI, Lab Populat Genet, Bethesda, MD 20892 USA. Univ Washington, Dept Genet, Seattle, WA 98195 USA. RP Washington Univ, Sch Med, Dept Genet, Campus Box 8232,4566 Scott Ave, St Louis, MO 63110 USA. EM ts@genetics.wustl.edu FU NICHD NIH HHS [R01 HD025614, HD25614]; NIGMS NIH HHS [GM15622] NR 66 TC 100 Z9 105 U1 0 U2 11 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0950-1991 EI 1477-9129 J9 DEVELOPMENT JI Development PD DEC PY 2000 VL 127 IS 24 BP 5265 EP 5276 PG 12 WC Developmental Biology SC Developmental Biology GA 388VY UT WOS:000166204800005 PM 11076749 ER PT J AU Hou, L Panthier, JJ Arnheiter, H AF Hou, L Panthier, JJ Arnheiter, H TI Signaling and transcriptional regulation in the neural crest-derived melanocyte lineage: interactions between KIT and MITF SO DEVELOPMENT LA English DT Article DE receptor tyrosine kinase; basic-helix-loop-helix-leucine zipper protein; dautochrome tautomerase; pMel 17/silver; tyrosinase-related protein-1; tyrosinase; cholera toxin; neural crest cell culture ID MICROPHTHALMIA GENE-PRODUCT; RECEPTOR TYROSINE KINASE; ENDOTHELIN-B RECEPTOR; STEM-CELL FACTOR; W-LOCUS; PROTO-ONCOGENE; MOUSE EMBRYO; MUTATIONS; EXPRESSION; DIFFERENTIATION AB Genetic and cell culture analyses have shown that the development of melanocytes from neural crest-derived precursor cells critically depends on the tyrosine kinase receptor KIT and the basic-helix-loop-helix-leucine zipper transcription factor MITF, KIT and MITF show complex interactions in that MITF is needed for the maintenance of Kit expression in melanoblasts and KIT signaling modulates MTF activity and stability in melanocyte cell lines, Using primary neural crest cell cultures from embryos homozygous for a Kit null allele marked by an inserted LacZ gene (Kit(W-LacZ)), show that the onset of Mitf expression in melanoblasts does not require KIT, In fact, provided that the melanocyte growth factor endothelin-3 is present, a small number of MITF/beta -Gal-positive cells can be maintained for at least 2 weeks in Kit(W-LacZ)/Kit(W-LacZ) cultures. These cells express several pigment cell-specific genes that are thought or have been shown to be activated by MITF, including dautochrome tautomerase, pMel 17/Silver and tyrosinase-related protein-1, but lack expression of the MITF target gene tyrosinase, which encodes the rate-limiting enzyme in melanin synthesis. Consequently, the cells remain unpigmented, Addition of cholera toxin, which elevates cAMP levels and mimics part of the KIT signaling pathway, increases the number of MITF-positive cells in Kit(W-LacZ)/Kit(W-LacZ) cultures, leads to tyrosinase expression, and induces the differentiation of melanoblasts into mature, pigmented melanocytes, Even when added on day 5-6 of culture, cholera toxin still rescues tyrosinase expression and differentiation. The results thus demonstrate that the presence of MITF is not sufficient for tyrosinase expression in melanoblasts and that KIT signaling influences gene expression during melanocyte. development in a gene-selective manner. C1 NINDS, Lab Dev Neurogenet, NIH, Bethesda, MD 20892 USA. Ecole Natl Vet Alfort, UMR 955 INRA, Maisons Alfort, France. RP Arnheiter, H (reprint author), NINDS, Lab Dev Neurogenet, NIH, Bethesda, MD 20892 USA. RI PANTHIER, Jean-Jacques/I-4366-2014; OI PANTHIER, Jean-Jacques/0000-0002-7966-0663; Hou, Ling/0000-0003-0705-8099 NR 55 TC 107 Z9 110 U1 0 U2 3 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0950-1991 J9 DEVELOPMENT JI Development PD DEC PY 2000 VL 127 IS 24 BP 5379 EP 5389 PG 11 WC Developmental Biology SC Developmental Biology GA 388VY UT WOS:000166204800015 PM 11076759 ER PT J AU Denham, SA Workman, E Cole, PM Weissbrod, C Kendziora, KT Zahn-Waxler, C AF Denham, SA Workman, E Cole, PM Weissbrod, C Kendziora, KT Zahn-Waxler, C TI Prediction of externalizing behavior problems from early to middle childhood: The role of parental socialization and emotion expression SO DEVELOPMENT AND PSYCHOPATHOLOGY LA English DT Article ID MANAGE PRESCHOOL BOYS; REFERRED PROBLEM 3-YEAR-OLDS; FAMILY-INTERACTION PATTERNS; CHILDRENS CONDUCT PROBLEMS; FOLLOW-UP; REARING PRACTICES; COMPETENCE; PERSPECTIVE; DELINQUENCY; ADOLESCENTS AB Parental emotions and behaviors that contribute to continuity and change in preschool children's externalizing problems were examined. Mothers and fathers were observed interacting with their children, and child-rearing styles were reported. Teachers, mothers, and children reported children's antisocial, oppositional behavior. Externalizing problems showed strong continuity 2 and 4 years later. Proactive parenting (i.e., supportive presence, clear instruction, and limit setting) predicted fewer behavior problems over time, after controlling for initial problems; the converse was true for parental anger. In contrast, the hypothesized ameliorative contribution of parents' positive emotion was not found. Parental contributions were most influential for children whose initial problems were in the clinical range. In particular, parental anger predicted continuation of problems over time. Paternal, as well as maternal, influences were identified. Examination of parental emotions and inclusion of fathers is important to research and intervention with young antisocial children. C1 George Mason Univ, Dept Psychol, Fairfax, VA 22030 USA. NIMH, Sect Dev Psychopathol, Bethesda, MD 20892 USA. Penn State Univ, University Pk, PA 16802 USA. American Univ, Washington, DC 20016 USA. RP Denham, SA (reprint author), George Mason Univ, Dept Psychol, 4400 Univ Dr,Mailstop 3F5, Fairfax, VA 22030 USA. NR 73 TC 194 Z9 198 U1 7 U2 61 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0954-5794 J9 DEV PSYCHOPATHOL JI Dev. Psychopathol. PD WIN PY 2000 VL 12 IS 1 BP 23 EP 45 DI 10.1017/S0954579400001024 PG 23 WC Psychology, Developmental SC Psychology GA 301AJ UT WOS:000086285700002 PM 10774594 ER PT J AU Dasso, JF Obiakor, H Bach, H Anderson, AO Mage, RG AF Dasso, JF Obiakor, H Bach, H Anderson, AO Mage, RG TI A morphological and immunohistological study of the human and rabbit appendix for comparison with the avian bursa SO DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY LA English DT Article DE human appendix; rabbit appendix; lymphoid development; chicken bursa; germinal center; immunohistochemistry; T-cell; B-cell repertoire ID B-CELL REPERTOIRE; SOMATIC DIVERSIFICATION; GENE CONVERSION; TONSILLECTOMIZED CHILDREN; IMMUNE-SYSTEM; V-H; CHICKEN; HYPERMUTATION; SEQUENCES; FABRICIUS AB Diversification of the primary antibody repertoire occurs in young rabbit appendix, As a prelude to molecular investigation of whether human appendix has a similar role, we compared the lymphoid morphology and distribution of common B- anal T-cell subsets in frozen and/or parafiin-embedded normal appendix specimens at various ages, IgA, IgM and IgG staining patterns were similar in frozen human and rabbit appendices. The elongated follicles of the young human and rabbit appendices regressed with age to resemble Peyer's patches. Although similar in morphology to the bursa, human and rabbit appendix follicles differ in that they do not involute completely with age and contain significant numbers of germinal center (GC) T cells although the number is low early in life. If the human appendix functions as a primary lymphoid organ. it may occur during the first few months of age when the GC T-cell density is low. Published by Elsevier Science Ltd. C1 NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. USA, Med Res Inst Infect Dis, Frederick, MD 21702 USA. RP Dasso, JF (reprint author), NIAID, Immunol Lab, NIH, Bldg 10,Room 11N311,10 Ctr Dr MSC 1892, Bethesda, MD 20892 USA. NR 60 TC 29 Z9 32 U1 1 U2 12 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0145-305X J9 DEV COMP IMMUNOL JI Dev. Comp. Immunol. PD DEC PY 2000 VL 24 IS 8 BP 797 EP 814 DI 10.1016/S0145-305X(00)00033-1 PG 18 WC Immunology; Zoology SC Immunology; Zoology GA 338HM UT WOS:000088413300008 PM 10906392 ER PT J AU Robb, L Hartley, L Begley, CG Brodnicki, TC Copeland, NG Gilbert, DJ Jenkins, NA Elefanty, AG AF Robb, L Hartley, L Begley, CG Brodnicki, TC Copeland, NG Gilbert, DJ Jenkins, NA Elefanty, AG TI Cloning, expression analysis, and chromosomal localization of murine and human homologues of a Xenopus Mix gene SO DEVELOPMENTAL DYNAMICS LA English DT Article DE embryogenesis; gastrulation; primitive streak; mesoderm; Mm1; Mix.1; Mix.2; Bix; CMIX; mixer; tail-bud; embryoid body ID ANTERIOR PRIMITIVE ENDODERM; TGF-BETA SUPERFAMILY; HOMEOBOX GENE; MESODERM INDUCTION; HOMEODOMAIN PROTEINS; DNA-SEQUENCES; EMBRYOS; ACTIVIN; CELLS; ZEBRAFISH AB We report the cloning and chromosomal localization of murine and human Mix genes, members of a subclass of paired-like homeobox genes of which the Xenopus laevis Mix.1 gene is the founding member. The murine Mix gene was mapped to the distal region of chromosome 1 and the human region to the syntenic region 1q41-42. Northern analysis and RT-PCR of murine adult and embryonic tissues demonstrated that Mix expression was restricted to the early embryo. Whole-mount in situ hybridization revealed patchy but symmetrical Mix expression in visceral endoderm of embryonic day (E)5.5 embryos. In slightly older embryos, the expression was skewed to one side of the embryo and by E6.5, at the onset of gastrulation, expression was seen in the epiblast, visceral endoderm, nascent mesoderm, and the primitive streak. This expression pattern was maintained in mid- and late-streak embryos. In early bud-stage embryos, expression was strongest in the proximal two thirds of the streak, extending to the base of the allantois. By the headfold-stage, expression was confined to the remnant of the primitive streak in the caudal region of the embryo and, after E8.0, in the caudal notochord and tail bud mesoderm. Mix transcripts were no longer detectable after embryonic day 9.5. (C) 2000 Wiley-Liss, Inc. C1 Walter & Eliza Hall Inst Med Res, Rotary Bone Marrow Res Labs, Melbourne, Vic 3050, Australia. NCI, Mammalian Genet Lab, ABL Basic Res Program, Frederick Canc Res & Dev Ctr, Frederick, MD 21701 USA. RP Robb, L (reprint author), Walter & Eliza Hall Inst Med Res, Rotary Bone Marrow Res Labs, PO Royal Melbourne Hosp, Melbourne, Vic 3050, Australia. RI Elefanty, Andrew/A-6066-2008 NR 39 TC 60 Z9 63 U1 0 U2 2 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 1058-8388 J9 DEV DYNAM JI Dev. Dyn. PD DEC PY 2000 VL 219 IS 4 BP 497 EP 504 DI 10.1002/1097-0177(2000)9999:9999<::AID-DVDY1070>3.0.CO;2-O PG 8 WC Anatomy & Morphology; Developmental Biology SC Anatomy & Morphology; Developmental Biology GA 377EE UT WOS:000165498300004 PM 11084649 ER PT J AU SanGiovanni, JP Allred, EN Mayer, DL Stewart, JE Herrera, MG Leviton, A AF SanGiovanni, JP Allred, EN Mayer, DL Stewart, JE Herrera, MG Leviton, A TI Reduced visual resolution acuity and cerebral white matter damage in very-low birthweight infants SO DEVELOPMENTAL MEDICINE AND CHILD NEUROLOGY LA English DT Article ID LOW-BIRTH-WEIGHT; INTRAVENTRICULAR HEMORRHAGE; MONOCULAR ACUITY; CARD PROCEDURE; CHILDREN; MRI; ULTRASOUND; BRAIN; LEUKOMALACIA; IMPAIRMENT AB Neonatal cerebral white matter echolucencies predict visual resolution acuity deficits in very-low-birthweight (VLBW) infants. We examined maternal sociodemographic, lifestyle, intrapartum, infant birth/perinatal, and ocular motor/refractive characteristics to determine whether they accounted for this association in infants who were tested once between postnatal age 25 and 56 weeks (corrected for gestational age at birth). Granial ultrasound scans were read by consensus to identify echolucency in a population of VLBW infants with no known ocular abnormalities. Visual resolution acuity was measured with the Acuity card Procedure (ACP) in 14 infants with echolucency and compared with that of 81 VLBW infants born in the same hospitals with normal ultrasound scans. Is time-oriented logistic regression models, echolucency remained a consistent predictor of abnormal visual resolution acuity after adjustment for covariates in three developmental periods (pre-, peri-, and postnatal). Odds ratios ranged from 19.3 (95% confidence interval, 4.5 to 82.2;p=0.001) to 10.4 (95% confidence interval, 1.3 to 81.9;p=0.03). Reduced visual resolution acuity in VLBW infants appears to be due to cerebral white matter damage. C1 NEI, Div Epidemiol & Clin Res, NIH, Bethesda, MD 20892 USA. Childrens Hosp, Dept Neurol, Neuroepidemiol Unit, Boston, MA 02115 USA. Childrens Hosp, Dept Ophthalmol, Boston, MA 02115 USA. Beth Israel Deaconess Med Ctr, Dept Neonatol, Boston, MA 02215 USA. Harvard Univ, Sch Publ Hlth, Dept Nutr, Boston, MA 02115 USA. RP SanGiovanni, JP (reprint author), NEI, Div Epidemiol & Clin Res, NIH, Bldg 131,31 Ctr Dr,MSC 2510, Bethesda, MD 20892 USA. RI SanGiovanni, John Paul/A-7605-2008 FU NINDS NIH HHS [NS 27306] NR 38 TC 10 Z9 10 U1 0 U2 0 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 USA SN 0012-1622 J9 DEV MED CHILD NEUROL JI Dev. Med. Child Neurol. PD DEC PY 2000 VL 42 IS 12 BP 809 EP 815 PG 7 WC Clinical Neurology; Pediatrics SC Neurosciences & Neurology; Pediatrics GA 384YQ UT WOS:000165975200004 PM 11132254 ER PT J AU Weyer, C Hanson, RL Tataranni, PA Bogardus, C Pratley, RE AF Weyer, C Hanson, RL Tataranni, PA Bogardus, C Pratley, RE TI A high fasting plasma insulin concentration predicts type 2 diabetes independent of insulin resistance - Evidence for a pathogenic role of relative hyperinsulinemia SO DIABETES LA English DT Article ID JAPANESE-AMERICAN MEN; BODY-WEIGHT GAIN; PIMA-INDIANS; BETA-CELL; GLUCOSE-TRANSPORTER; NATURAL-HISTORY; FOLLOW-UP; SECRETORY DYSFUNCTION; RISK-FACTORS; NIDDM AB Fasting hyperinsulinemia is a widely used surrogate measure of insulin resistance and predicts type 2 diabetes in various populations, whether fasting hyperinsulinemia predicts diabetes independent of insulin resistance is unknown. In 319 Pima Indians with. normal glucose tolerance, fasting plasma insulin concentration and insulin-stimulated glucose disposal (M) (hyperinsulinemic clamp) were inversely related, but at any given M, there was substantial variation, with some subjects being hyperinsulinemic and others being hypoinsulinemic relative to their degree of insulin sensitivity. In 262 of the 319 subjects followed prospectively over 6.4 +/- 3.9 years, a high fasting plasma insulin concentration was a significant independent predictor of diabetes, in addition to low M and low acute insulin response (AIR) (intravenous glucose challenge). In 161 of the 319 subjects with follow-up measurements of M and AIR (5.1 +/- 3.9 years), a high relative fasting plasma insulin concentration predicted a decline in AIR but not in M before the onset of diabetes. The adjusted fasting plasma insulin concentration was a familial trait (heritability of 0.52) and in a genome-wide scan, there was suggestive evidence of linkage (logarithm of odds score 1.77) to a region on chromosome 3q, which harbors the gene encoding GLUT2. These results provide the first prospective evidence in humans that fasting hyperinsulinemia itself has a primary role in the pathogenesis of diabetes, independent of insulin resistance. Whether amelioration of basal insulin hypersecretion will prevent diabetes remains to be elucidated. C1 NIDDKD, Clin Diabet & Nutr Sect, NIH, Phoenix, AZ 85016 USA. RP Weyer, C (reprint author), NIDDKD, Clin Diabet & Nutr Sect, NIH, 4212 N 16th St,Rm 5-41, Phoenix, AZ 85016 USA. RI Hanson, Robert/O-3238-2015 OI Hanson, Robert/0000-0002-4252-7068 NR 55 TC 133 Z9 139 U1 0 U2 4 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0012-1797 J9 DIABETES JI Diabetes PD DEC PY 2000 VL 49 IS 12 BP 2094 EP 2101 DI 10.2337/diabetes.49.12.2094 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 378FC UT WOS:000165573500017 PM 11118012 ER PT J AU Meigs, JB Cupples, LA Wilson, PWF AF Meigs, JB Cupples, LA Wilson, PWF TI Parental transmission of type 2 diabetes - The Framingham Offspring Study SO DIABETES LA English DT Article ID IMPAIRED GLUCOSE-TOLERANCE; MATERNAL HISTORY; UNITED-STATES; NIDDM; MELLITUS; PREVALENCE; DISEASE; EXCESS; FAMILIES; OBESITY AB Study of parental transmission of diabetes provides insight into the relative contributions of underlying maternal and paternal influences. We estimated risk for type 2 diabetes and milder degrees of glucose intolerance associated with parental diabetes among subjects of the population-based Framingham Offspring Study, in which participants are primarily Caucasian and at relatively low risk for diabetes and for which both parental and offspring phenotypes were ascertained by direct examination. Parental diabetes, assessed over 40 years of biennial follow-up, was defined by use of hypoglycemic drug therapy or a casual plasma glucose level greater than or equal to 11.1 mmol/l at any examination. Offspring glucose tolerance, assessed over 20 years of quadrennial follow-up, was defined by fasting plasma glucose levels greater than or equal to7.8 mmol/l at any two examinations, use of hypoglycemic drug therapy at any examination, or with a 75-g oral glucose tolerance test (1980 World Health Organization criteria) at the most recent examination. We calculated odds ratios (ORs) and 95% CIs for offspring glucose tolerance status using generalized estimating equations to account fdr differential correlations within and between families. The 2,527 offspring came from 1,303 nuclear families, of which 77.6% had two or more siblings per family and in which the prevalence of parental diabetes was 24.6%. The mean offspring age was 54 years (range 26-82), 53% were women, 8.6% had diabetes, 11.4% had impaired glucose tolerance, 76.3% had no parental diabetes, 10.5% had maternal diabetes, 11.5% had paternal diabetes, and 1.7% had bilineal diabetes. Relative to individuals without parental diabetes, the age-adjusted ORs (95% CI) for offspring type 2 diabetes or abnormal glucose tolerance (fasting plasma glucose greater than or equal to6.1 mmol/l or. 2-h postchallenge glucose tolerance greater than or equal to7,8 mmol/l) among individuals with maternal diabetes were 3.4 (2.3-4.9) and 2.7 (2.0-3.7), respectively; among individuals with paternal diabetes were 3.5 (2.3-5.2) and 1.7 (1.2-2.4), respectively; and among individuals with bilineal diabetes were 6.1 (2.9-13.0) and 5.2 (2.6-10.5), respectively. Although maternal and paternal diabetes conferred equivalent risk for offspring type 2 diabetes, offspring with maternal diabetes were slightly more likely to have abnormal glucose tolerance compared with those with paternal diabetes (OR 1.6, 95% CI 1.1-2.4). Offspring with maternal diabetes and an age Of onset of <50 years had marked increased risk for both type 2 diabetes (9.7, 4.3-22.0) and abnormal glucose tolerance (9.0, 4.2-19.7). We conclude that risk ratios for offspring type 2 diabetes are consistent with a simple additive risk model, where risk when both parents are affected equals the sum of risk when either parent is affected. For maternal diabetes to confer excess risk for mild but not overt glucose intolerance, offspring of diabetic fathers may transit abnormal to impaired glucose tolerance relatively quickly, or diabetic mothers may transmit risk for a mild slowly progressive form of abnormal glucose tolerance in addition to overt diabetes. Very high risk for abnormal glucose homeostasis among offspring with young age-of-onset maternal diabetes is consistent with hypotheses that perinatal exposures increase diabetes risk. Given equivalent risk ratios for type 2 diabetes, fathers may transmit unique paternal genetic factors of similar strength to maternal environmental factors. C1 Massachusetts Gen Hosp, Gen Internal Med Unit, Boston, MA 02114 USA. Massachusetts Gen Hosp, Div Gen Med, Boston, MA 02114 USA. Massachusetts Gen Hosp, Dept Med, Boston, MA 02114 USA. Harvard Univ, Sch Med, Boston, MA USA. Boston Univ, Sch Publ Hlth, Dept Epidemiol & Biostat, Boston, MA USA. Boston Univ, Sch Med, Boston, MA 02118 USA. NHLBI, Framingham Heart Study, Framingham, MA USA. RP Meigs, JB (reprint author), Harvard Univ, Massachusetts Gen Hosp, Gen Internal Med Unit S50 9, 50 Staniford St,9th Floor, Boston, MA 02114 USA. FU NHLBI NIH HHS [N01-HC-38083] NR 43 TC 250 Z9 258 U1 3 U2 11 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0012-1797 J9 DIABETES JI Diabetes PD DEC PY 2000 VL 49 IS 12 BP 2201 EP 2207 DI 10.2337/diabetes.49.12.2201 PG 7 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 378FC UT WOS:000165573500031 PM 11118026 ER PT J AU Dabelea, D Hanson, RL Lindsay, RS Pettitt, DJ Imperatore, G Gabir, MM Roumain, J Bennett, PH Knowler, WC AF Dabelea, D Hanson, RL Lindsay, RS Pettitt, DJ Imperatore, G Gabir, MM Roumain, J Bennett, PH Knowler, WC TI Intrauterine exposure to diabetes conveys risks for type 2 diabetes and obesity - A study of discordant sibships SO DIABETES LA English DT Article ID PIMA INDIAN WOMEN; INSULIN-SECRETION; ADULT-RATS; MELLITUS; MOTHERS; PREGNANCY; HYPERGLYCEMIA; INHERITANCE; PREVALENCE; CHILDREN AB Intrauterine exposure to diabetes is associated with an excess of diabetes and obesity in the offspring, but the effects of intrauterine exposure are confounded by genetic factors. To determine the role of the intrauterine diabetic environment per se, the prevalence of diabetes and the mean BMI were compared in siblings born before and after their mother was recognized as having diabetes. Nuclear families in which at least one sibling was born before and one after the mother was diagnosed with type 2 diabetes were selected. Consequently, the siblings born before and after differed in their exposure to diabetes in utero. A total of 58 siblings from 19 families in which at least one sibling had diabetes were examined at similar ages (within 3 years). The risk of diabetes was significantly higher in siblings born after the mother developed diabetes than in those born before the mother's diagnosis of diabetes (odds ratio 3.7, P = 0.02). In 52 families, among 183 siblings without diabetes, the mean BMI was 2.6 kg/m(2) higher in offspring of diabetic than in offspring of nondiabetic pregnancies (P = 0.003). In contrast, there were no significant differences in risk of diabetes or BMI between offspring born before and after the father was diagnosed with diabetes. Intrauterine exposure to diabetes per se conveys a high risk for the development of diabetes and obesity in offspring in excess of risk attributable to genetic factors alone. C1 NIDDKD, Diabet & Arthrit Epidemiol Sect, NIH, Phoenix, AZ 85014 USA. RP Knowler, WC (reprint author), NIDDKD, Diabet & Arthrit Epidemiol Sect, NIH, 1550 E Indian Sch Rd, Phoenix, AZ 85014 USA. RI Hanson, Robert/O-3238-2015 OI Hanson, Robert/0000-0002-4252-7068 NR 30 TC 537 Z9 551 U1 7 U2 28 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0012-1797 J9 DIABETES JI Diabetes PD DEC PY 2000 VL 49 IS 12 BP 2208 EP 2211 DI 10.2337/diabetes.49.12.2208 PG 4 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 378FC UT WOS:000165573500032 PM 11118027 ER PT J AU Weyer, C Foley, JE Bogardus, C Tataranni, PA Pratley, RE AF Weyer, C Foley, JE Bogardus, C Tataranni, PA Pratley, RE TI Enlarged subcutaneous abdominal adipocyte size, but not obesity itself, predicts Type II diabetes independent of insulin resistance SO DIABETOLOGIA LA English DT Article DE obesity; adipose tissue; glucose intolerance; predictors; prospective study ID HUMAN ADIPOSE-TISSUE; BODY-FAT DISTRIBUTION; PIMA-INDIANS; CELL SIZE; SECRETORY DYSFUNCTION; GROWTH-HORMONE; RISK-FACTORS; PPAR-GAMMA; MELLITUS; GLUCOSE AB Aims/hypothesis. Cross-sectional studies indicate that enlarged subcutaneous abdominal adipocyte size is associated with hyperinsulinaemia, insulin resistance and glucose intolerance. To further explore the pathophysiological significance of these associations, we examined prospectively whether enlarged subcutaneous abdominal adipocyte size predicts Type II (non-insulin-dependent) diabetes mellitus. Methods. Body composition (hydrodensitometry), mean subcutaneous abdominal adipocyte size (fat biopsy), insulin sensitivity (hyperinsulinaemic clamp) and the acute insulin secretory response (25-g i.v. GTT) were assessed in 280 Pima Indians with either normal (NGT), impaired (IGT) or diabetic glucose tolerance (75-g OGTT). Subjects with NGT were then followed prospectively. Results. After adjusting for age, sex and per cent body fat, mean subcutaneous abdominal adipocyte size was 19% and 11% higher in subjects with diabetes and IGT, compared with those with NGT (p < 0.001). Insulin sensitivity was inversely correlated with mean subcutaneous abdominal adipocyte size (r = -0.53, p < 0.0001), even after adjusting for per cent body fat (r = -0.31, p < 0.001). In 108 NGT subjects followed over 9.3 +/- 4.1 years (33 of whom developed diabetes), enlarged mean subcutaneous abdominal adipocyte size but not high per cent body fat, was an independent predictor of diabetes, in addition to a low insulin sensitivity and acute insulin secretory response [relative hazard 10(th) vs 90(th) centile (95% CI): 5.8 (1.7-19.6), p < 0.005]. In 28 NGT subjects with a 9% weight gain over 2.7 +/- 1.3 years, changes in insulin sensitivity were inversely and independently related to changes in mean subcutaneous abdominal adipocyte size and per cent body fat. Conclusion/interpretation. Although enlarged mean subcutaneous abdominal adipocyte size is associated with insulin resistance cross-sectionally, prospectively, both abnormalities are independent and additive predictors of Type II diabetes. C1 NIDDKD, Clin Diabet & Nutr Sect, NIH, Phoenix, AZ 85016 USA. RP Weyer, C (reprint author), NIDDKD, Clin Diabet & Nutr Sect, NIH, 4212 N 16th St,Rm 5-14, Phoenix, AZ 85016 USA. NR 61 TC 345 Z9 356 U1 2 U2 11 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0012-186X J9 DIABETOLOGIA JI Diabetologia PD DEC PY 2000 VL 43 IS 12 BP 1498 EP 1506 DI 10.1007/s001250051560 PG 9 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 381GT UT WOS:000165754500006 PM 11151758 ER PT J AU Bears, M Cartwright, R Mercer, P AF Bears, M Cartwright, R Mercer, P TI Masochistic dreams: A gender-related diathesis for depression revisited SO DREAMING LA English DT Article DE dreaming; depression; gender role; gender; REM sleep ID REM-SLEEP AB Masochistic dreams, as defined by Beck (1967), ave reportedly more prevalent among women and individuals with past or present depression. However it is unclear whether these prevalence differences are a function of depressogenic personality traits or functioning mood symptoms. In the present study, 30 men and 30 women without histories of major depression slept two consecutive nights in a sleep laboratory and reported their dreams from each REM period on the second night. Dream content from this sample was compared to that of 60 depressed participants who were studied previously under the same protocol. Analyses did not support a heightened prevalence of masochistic dreams among women or depressed individuals. interestingly, the masochistic dreams of the non-depressed sample were equally distributed across the night, whereas depressed individuals tend to report masochistic dreams closer to morning. This hypothesized pattern suggests that masochistic dreams may be pathognomic of depression in that their occurrence near the end of the night affects morning mood with negative dream residue. C1 Rush Presbyterian St Lukes Med Ctr, Sleep Disorder Serv, Chicago, IL 60612 USA. Rush Presbyterian St Lukes Med Ctr, Res Ctr, Dept Psychol, Chicago, IL 60612 USA. RP Bears, M (reprint author), Univ Florida, Ctr Study Emot & Attent, NIMH Ctr, POB 100165, Gainesville, FL 32610 USA. NR 21 TC 3 Z9 3 U1 1 U2 2 PU KLUWER ACADEMIC-HUMAN SCIENCES PRESS PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013-1578 USA SN 1053-0797 J9 DREAMING JI Dreaming PD DEC PY 2000 VL 10 IS 4 BP 211 EP 219 DI 10.1023/A:1009480907418 PG 9 WC Psychology, Multidisciplinary SC Psychology GA 370LB UT WOS:000165123600003 ER PT J AU Joshi, BH Husain, SR Puri, RK AF Joshi, BH Husain, SR Puri, RK TI Preclinical studies with IL-13PE38QQR for therapy of malignant glioma SO DRUG NEWS & PERSPECTIVES LA English DT Article ID ENDOTHELIAL GROWTH-FACTOR; HUMAN B-CELLS; PSEUDOMONAS EXOTOXIN; INTERLEUKIN-13 RECEPTOR; CHIMERIC PROTEIN; BINDING SUBUNIT; IL-13 RECEPTOR; UP-REGULATION; GAMMA-CHAIN; EXPRESSION AB To develop novel therapeutic agents for the treatment of brain tumors, we have been investigating the expression of unique tumor-associated receptors or antigens on the tumor cell surface. About six years ago, we discovered that human solid tumor cell lines, including human malignant glioma, express high- to intermediate-affinity receptors (R) for a Th2 cell-derived cytokine, interleukin-13 (IL-13). Analysis of the subunit composition of IL-13R in primary explants of malignant glioma cells has demonstrated that IL-13R is composed of three different chains (IL-13R alpha1, IL-13R alpha2 and IL-4R alpha, also known as IL-13R alpha', alpha and IL-4R beta respectively) and that IL-13R alpha2 chain is overexpressed on these cells. Normal brain tissues express IL-13R alpha1 and IL-4R alpha chains, but show only marginal expression of IL-13R alpha2 chain. Thus IL-13R alpha2 chain appears to be overex pressed on glioma cells and may serve as a novel tumor biomarker or a target for receptor-directed therapeutic agents for brain tumors. To target IL-13 receptors, we have produced a recombinant fusion protein composed of IL-13 and a mutated form of Pseudomonas exotoxin (PE). This cytotoxin, termed IL-13PE38QQR or IL-13 cytotoxin, is highly and specifically cytotoxic to a spectrum of human glioma cell lines. In preclinical models of human glioblastoma tumors growing subcutaneously in immunodeficient mice, IL-13 cytotoxin has been found to have remarkable antitumor activity. The data that emerged from these studies reveal that localized or systemic administration of IL-13 cytotoxin can produce nontoxic drug levels and that IL-13 cytotoxin is potently effective against established glioblastoma tumors. On the basis of these and other preclinical studies, we have begun a phase I clinical trial using IL-13PE38QQR for therapy of recurrent malignant glioma. (C) 2000 Prous Science. All rights reserved. C1 US FDA, Lab Mol Tumor Biol, Div Cellular & Gene Therapies, Ctr Biol Evaluat & Res,NIH, Bethesda, MD 20892 USA. US FDA, Lab Mol Tumor Biol, Div Cellular & Gene Therapies, Ctr Biol Evaluat & Res,NIH, Bethesda, MD 20892 USA. NR 56 TC 10 Z9 10 U1 0 U2 1 PU PROUS SCIENCE, SA PI BARCELONA PA PO BOX 540, PROVENZA 388, 08025 BARCELONA, SPAIN SN 0214-0934 J9 DRUG NEWS PERSPECT JI Drug News Perspect. PD DEC PY 2000 VL 13 IS 10 BP 599 EP 605 DI 10.1358/dnp.2000.13.10.858450 PG 7 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 412PY UT WOS:000167564600004 PM 12879131 ER PT J AU Schmalzing, D Buonocore, S Piggee, C AF Schmalzing, D Buonocore, S Piggee, C TI Capillary electrophoresis-based immunoassays SO ELECTROPHORESIS LA English DT Review DE capillary electrophoresis; immunoassays; microchips; review ID LASER-INDUCED FLUORESCENCE; COMPETITIVE IMMUNOASSAY; MONOCLONAL-ANTIBODY; LABELED PEPTIDES; PROTEIN; POLARIZATION; SEPARATION; METHAMPHETAMINE; CONFIRMATION; SYSTEMS AB This review covers the progress and developments in the field of capillary electrophoresis immunoassay (CEIA) over the past three years. Because many excellent descriptions of the principles of these methods are available (e.g., in the reviews listed in this article), no elementary introduction is given to the field of immunoassays (IAs) or CEIAs. This report focuses exclusively on experimental results, dividing the CEIA papers into the categories of direct, indirect, and microchip electrophoretic immunoassays. in the last section, a brief summary of the current status of the CEIA field is presented. C1 Whitehead Inst Biomed Res, Cambridge Ctr 9, Cambridge, MA 02142 USA. NIMH, Lab Neurotoxicol, NIH, Bethesda, MD 20892 USA. RP Schmalzing, D (reprint author), Whitehead Inst Biomed Res, Cambridge Ctr 9, Cambridge, MA 02142 USA. NR 43 TC 60 Z9 61 U1 1 U2 9 PU WILEY-V C H VERLAG GMBH PI BERLIN PA PO BOX 10 11 61, D-69451 BERLIN, GERMANY SN 0173-0835 J9 ELECTROPHORESIS JI Electrophoresis PD DEC PY 2000 VL 21 IS 18 BP 3919 EP 3930 DI 10.1002/1522-2683(200012)21:18<3919::AID-ELPS3919>3.0.CO;2-F PG 12 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 387CN UT WOS:000166102700005 PM 11192116 ER PT J AU Wolfgang, M van Putten, JPM Hayes, SF Dorward, D Koomey, M AF Wolfgang, M van Putten, JPM Hayes, SF Dorward, D Koomey, M TI Components and dynamics of fiber formation define a ubiquitous biogenesis pathway for bacterial pili SO EMBO JOURNAL LA English DT Article DE general secretion pathway; pilus retraction; secretin; twitching motility; type IV pili ID EXTRACELLULAR PROTEIN SECRETION; TWITCHING MOTILITY GENE; SOCIAL GLIDING MOTILITY; NEISSERIA-GONORRHOEAE; PSEUDOMONAS-AERUGINOSA; OUTER-MEMBRANE; IV PILI; CHAPERONE/USHER PATHWAY; NATURAL TRANSFORMATION; MYXOCOCCUS-XANTHUS AB Type IV pili (Tfp) are a unique class of multifunctional surface organelles in Gram-negative bacteria, which play important roles in prokaryotic cell biology. Although components of the Tfp biogenesis machinery have been characterized, it is not clear how they function or interact. Using Neisseria gonorrhoeae as a model system, we report here that organelle biogenesis can be resolved into two discrete steps: fiber formation and translocation of the fiber to the cell surface. This conclusion is based on the capturing of an intermediate state in which the organelle is retained within the cell owing to the simultaneous absence of the secretin family member and biogenesis component PilQ and the twitching motility/pilus retraction protein PilT. This finding is the first demonstration of a specific translocation defect associated with loss of secretin function, and additionally confirms the role of PilT as a conditional antagonist of stable pilus fiber formation. These findings have important implications for Tfp structure and function and are pertinent to other membrane translocation systems that utilize a highly related set of components. C1 Univ Oslo, Biotechnol Ctr Oslo, N-0316 Oslo, Norway. Univ Oslo, Inst Pharm, N-0316 Oslo, Norway. Univ Michigan, Sch Med, Dept Microbiol & Immunol, Ann Arbor, MI 48109 USA. NIAID, Rocky Mt Labs, Microbial Struct & Funct Lab, NIH, Hamilton, MT 59840 USA. NIAID, Rocky Mt Labs, Microscopy Branch, NIH, Hamilton, MT 59840 USA. RP Koomey, M (reprint author), Univ Oslo, Biotechnol Ctr Oslo, N-0316 Oslo, Norway. OI van Putten, Jos/0000-0002-4126-8172 FU NIAID NIH HHS [AI27837]; NIGMS NIH HHS [2 T32 GM07315] NR 56 TC 189 Z9 193 U1 0 U2 10 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD DEC 1 PY 2000 VL 19 IS 23 BP 6408 EP 6418 DI 10.1093/emboj/19.23.6408 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 381LF UT WOS:000165763800014 PM 11101514 ER PT J AU Sattlegger, E Hinnebusch, AG AF Sattlegger, E Hinnebusch, AG TI Separate domains in GCN1 for binding protein kinase GCN2 and ribosomes are required for GCN2 activation in amino acid-starved cells SO EMBO JOURNAL LA English DT Article DE eIF2 alpha kinase; GCN20; paromomycin; ribosomal A-site; translational control ID INITIATION FACTOR-II; SACCHAROMYCES-CEREVISIAE; EIF-2-ALPHA KINASE; TRANSFER-RNA; TRANSLATIONAL ACTIVATOR; DROSOPHILA-MELANOGASTER; PHENOTYPIC SUPPRESSION; EIF2-ALPHA KINASE; YEAST GCN2; HOMOLOG AB GCN2 stimulates GCN4 translation in amino acid-starved cells by phosphorylating the alpha -subunit of translation initiation factor 2. GCN2 function in vivo requires the GCN1/GCN20 complex, which binds to the N-terminal domain of GCN2, A C-terminal segment of GCN1 (residues 2052-2428) was found to be necessary and sufficient for binding GCN2 in vivo and in vitro. Overexpression of this fragment in wild-type cells impaired association of GCN2 with native GCN1 and had a dominant Gcn(-) phenotype, dependent on Arg2259 in the GCN1 fragment. Substitution of Arg2259 with Ala in full-length GCN1 abolished complex formation with native GCN2 and destroyed GCN1 regulatory function. Consistently, the Gcn(-) phenotype of gcn1-R2259A, but not that of gcn1 Delta, was suppressed by overexpressing GCN2. These findings prove that GCN2 binding to the C-terminal domain of GCN1, dependent on Arg2259, is required for high level GCN2 function in who. GCN1 expression conferred sensitivity to paromomycin in a manner dependent on its ribosome binding domain, supporting the idea that GCN1 binds near the ribosomal acceptor site to promote GCN2 activation by uncharged tRNA. C1 NICHHD, Lab Eukaryot Gen Rehabil, NIH, Bethesda, MD 20892 USA. RP Hinnebusch, AG (reprint author), NICHHD, Lab Eukaryot Gen Rehabil, NIH, 6 Ctr Dr,Bldg 6A,Room B1A-13, Bethesda, MD 20892 USA. NR 43 TC 49 Z9 53 U1 1 U2 10 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD DEC 1 PY 2000 VL 19 IS 23 BP 6622 EP 6633 DI 10.1093/emboj/19.23.6622 PG 12 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 381LF UT WOS:000165763800034 PM 11101534 ER PT J AU Salomon, DS Bianco, C Ebert, AD Khan, NI De Santis, M Normanno, N Wechselberger, C Seno, M Williams, K Sanicola, M Foley, S Gullick, W Persico, G AF Salomon, DS Bianco, C Ebert, AD Khan, NI De Santis, M Normanno, N Wechselberger, C Seno, M Williams, K Sanicola, M Foley, S Gullick, W Persico, G TI The EGF-CFC family: novel epidermal growth factor-related proteins in development and cancer SO ENDOCRINE-RELATED CANCER LA English DT Review ID MAMMARY EPITHELIAL-CELLS; LEFT-RIGHT ASYMMETRY; ONE-EYED PINHEAD; LEFT-RIGHT AXIS; DIFFERENTIAL IMMUNOHISTOCHEMICAL DETECTION; MEDIATED MESODERM INDUCTION; GASTRULATING MOUSE EMBRYO; CERVICAL-CARCINOMA CELLS; HUMAN GASTRIC CARCINOMAS; BETA SIGNALING PATHWAYS AB The EGF-CFC gene family encodes a group of structurally related proteins that serve as important competence factors during early embryogenesis in Xenopus, zebrafish, mice and humans. This multigene family consists of Xenopus FRL-1, zebrafish one-eyed-pinhead (oep), mouse cripto (Cr-l) and cryptic, and human cripto (CR-1) and criptin. FRL-1, oep and mouse cripto are essential for the formation of mesoderm and endoderm and for correct establishment of the anterior/posterior axis. In addition, oep and cryptic are important for the establishment of left-right (UR) asymmetry, In zebrafish, there is strong genetic evidence that oep functions as an obligatory cc-factor for the correct signaling of a transforming growth factor-beta (TGF beta)-related gene, nodal, during gastrulation and during UR asymmetry development. Expression of Cr-l and cryptic is extinguished in the embryo after day 8 of gestation except for the developing heart where Cr-l expression is necessary for myocardial development. In the mouse, cryptic is not expressed in adult tissues whereas Cr-l is expressed at a low level in several different tissues including the mammary gland, In the mammary gland, expression of Cr-l in the ductal epithelial cells increases during pregnancy and lactation and immunoreactive and biologically active Cr-l protein can be detected in human milk. Overexpression of Cr-l in mouse mammary epithelial cells can facilitate their in vitro transformation and in vivo these Cr-l-transduced cells produce ductal hyperplasias in the mammary gland. Recombinant mouse or human cripto can enhance cell motility and branching morphogenesis in mammary epithelial cells and in some human tumor cells. These effects are accompanied by an epithelial-mesenchymal transition which is associated with a decrease in beta -catenin function and an increase in vimentin expression. Expression of cripto is increased several-fold in human colon, gastric, pancreatic and lung carcinomas and in a variety of different types of mouse and human breast carcinomas. More importantly, this increase can first be detected in premalignant lesions in some of these tissues. Although a specific receptor for the EGF-CFC proteins has not yet been identified, oep depends upon an activin-type RIIB and RIB receptor system that functions through Smad-e. Mouse and human cripto have been shown to activate a ras/raf/MAP kinase signaling pathway in mammary epithelial cells. Activation of phosphatidylinositol 3-kinase and Akt are also important for the ability of CR-1 to stimulate cell migration and to block lactogenic hormone-induced expression of p-casein and whey acidic protein. In mammary epithelial cells, part of these responses may depend on the ability of CR-1 to transactivate erb B-4 and/or fibroblast growth factor receptor 1 through an src-like tyrosine kinase. C1 NCI, Tumor Immunol & Biol Lab, Tumor Growth Factor Sect, NIH, Bethesda, MD 20892 USA. Free Univ Berlin, UK Benjamin Franklin, Frauenklin, D-12200 Berlin, Germany. Inst Oncol Sperimentale, Naples, Italy. Okayama Univ, Okayama, Japan. Biogen Inc, Cambridge, MA 02142 USA. Univ Kent, Dept Biol Sci, Canterbury, Kent, England. Int Inst Genet & Biofis, Naples, Italy. RP Salomon, DS (reprint author), NCI, Tumor Immunol & Biol Lab, Tumor Growth Factor Sect, NIH, Bethesda, MD 20892 USA. RI SENO, Masaharu /B-2092-2011; OI SENO, Masaharu /0000-0001-8547-6259; Normanno, Nicola/0000-0002-7158-2605 NR 218 TC 94 Z9 98 U1 0 U2 4 PU SOC ENDOCRINOLOGY PI BRISTOL PA 17/18 THE COURTYARD, WOODLANDS, BRADLEY STOKE, BRISTOL BS32 4NQ, ENGLAND SN 1351-0088 J9 ENDOCR-RELAT CANCER JI Endocr.-Relat. Cancer PD DEC PY 2000 VL 7 IS 4 BP 199 EP 226 DI 10.1677/erc.0.0070199 PG 28 WC Oncology; Endocrinology & Metabolism SC Oncology; Endocrinology & Metabolism GA 450LA UT WOS:000169743300001 PM 11174844 ER PT J AU Chen, JR Cheng, JG Shatzer, T Sewell, L Hernandez, L Stewart, CL AF Chen, JR Cheng, JG Shatzer, T Sewell, L Hernandez, L Stewart, CL TI Leukemia inhibitory factor can substitute for nidatory estrogen and is essential to inducing a receptive uterus for implantation but is not essential for subsequent embryogenesis SO ENDOCRINOLOGY LA English DT Article ID EPIDERMAL GROWTH-FACTOR; MOUSE UTERUS; BLASTOCYST IMPLANTATION; UTERINE EXPRESSION; SIGNALING PATHWAYS; HUMAN ENDOMETRIUM; MICE LACKING; EGF-RECEPTOR; FACTOR LIF; GENE AB A stage critical in mammalian development is embryo implantation. At this point, the blastocyst establishes a close interaction with the uterine tissues, a step necessary for its continued embryonic development. In many mammalian species, including man, uterine expression of the cytokine, leukemia inhibitory factor (LIF) is coincident with the onset of implantation and in mice LIF is essential to this process. The reasons for implantation failure have not been established. Here we show in LIF-deficient mice that up to the onset of implantation, changes in uterine cell proliferation, hormone levels, blastocyst localization, as well as expression of lactoferrin and Muc-1, do not differ from wild-types. However, the uterus fails to respond to the presence of embryos or to artificial stimuli by decidualizing. In mice, implantation and decidualization are induced by nidatory estrogen. We show that uterine expression of LIF is up-regulated by estrogen and LIF can replace nidatory estrogen at inducing both implantation and decidualization in ovariectomized mice. Implantation of LIF-deficient embryos in the LIF-deficient females, with normal development to term is rescued by ip injection of LIF. Transient expression of LIF on D4 of pregnancy is therefore only required to induce a state of receptivity in the uterus permitting embryo implantation and decidualization. LIF is neither required by the embryo for development nor for the maintenance of pregnancy. C1 NCI, Frederick Canc Res & Dev Ctr, Lab Canc & Dev Biol, ABL Basic Res Program, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, Sci Applicat Int Corp, Frederick, MD 21702 USA. RP Stewart, CL (reprint author), NCI, Frederick Canc Res & Dev Ctr, Lab Canc & Dev Biol, ABL Basic Res Program, POB B, Frederick, MD 21702 USA. FU PHS HHS [N01-C0-56000] NR 48 TC 153 Z9 175 U1 0 U2 5 PU ENDOCRINE SOC PI BETHESDA PA 4350 EAST WEST HIGHWAY SUITE 500, BETHESDA, MD 20814-4110 USA SN 0013-7227 J9 ENDOCRINOLOGY JI Endocrinology PD DEC PY 2000 VL 141 IS 12 BP 4365 EP 4372 DI 10.1210/en.141.12.4365 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 374TR UT WOS:000165360900006 PM 11108244 ER PT J AU Liu, JL Yakar, S LeRoith, D AF Liu, JL Yakar, S LeRoith, D TI Mice deficient in liver production of insulin-like growth factor I display sexual dimorphism in growth hormone-stimulated postnatal growth SO ENDOCRINOLOGY LA English DT Article ID GH-BINDING PROTEIN; RECEPTOR GENE; BODY GROWTH; MALE-RATS; IGF-I; SECRETION; EXPRESSION; MOUSE; HYPOPHYSECTOMY; PROFILES AB Insulin-Like growth factor I (IGF-I) is essential for normal intrauterine and postnatal growth and development. Using the Cre/loxP-induced conditional knockout system, we have established a liver-specific IGF-I-deficient (LID) mouse model. Circulating TGF-I levers were decreased by approximately 75% without any apparent effect on their growth and development. To determine the role of extra-hepatic IGF-I in GH-induced postnatal growth, we tested the effects of GH on growth rates in these mice. Female, but not male, LID mice displayed significantly accelerated growth rates in response to daily injections of GH for 5 weeks. The GH-induced peripubertal growth in female LID mice was not affected by ovariectomy, nor did castration change the growth pattern in male LID mice. Thus, factors other than gonadal steroids mediate this sexual dimorphism. We postulate that the sexual dimorphic response to GH observed in LID mice may be related to genetically programmed differences in GH secretion patterns. C1 NIDDK, CEB, NIH, Bethesda, MD 20892 USA. RP LeRoith, D (reprint author), NIDDK, CEB, NIH, Bldg 10,Room 8D12,10 Ctr Dr, Bethesda, MD 20892 USA. NR 41 TC 47 Z9 48 U1 1 U2 4 PU ENDOCRINE SOC PI BETHESDA PA 4350 EAST WEST HIGHWAY SUITE 500, BETHESDA, MD 20814-4110 USA SN 0013-7227 J9 ENDOCRINOLOGY JI Endocrinology PD DEC PY 2000 VL 141 IS 12 BP 4436 EP 4441 DI 10.1210/en.141.12.4436 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 374TR UT WOS:000165360900014 PM 11108252 ER PT J AU Moore, JP Wray, S AF Moore, JP Wray, S TI Luteinizing hormone-releasing hormone (LHRH) biosynthesis and secretion in embryonic LHRH neurons SO ENDOCRINOLOGY LA English DT Article ID HYPOTHALAMIC GT1-7 NEURONS; SLICE EXPLANT CULTURES; OLFACTORY PLACODE; GENE-EXPRESSION; MESSENGER-RNA; RHESUS-MONKEY; MOUSE; BRAIN; GABA; HYPOGONADISM AB Evidence indicates that LH-releasing hormone (LHRH) neurons can exhibit neuroendocrine secretory properties before entrance into the central nervous system. In this study, we evaluated LHRH biosynthesis and secretion in embryonic LHRH neurons maintained in nasal explants. Using ELISA and calcium imaging techniques, peptide content and single neuron activities mere examined. LHRH neurons maintained for 7-10 days in vitro mere found to possess a similar amount of LHRH/cell as the equivalent aged LHRH cells in vivo (postnatal day ). LHRH peptide was measured in the medium of these relatively young cultures, and 40 mM KCl stimulated a 4-fold increase in LHRH secretion. KCI enhanced medium also resulted in a significant increase in LHRH content per culture (24.5 pg vs. 32.3). A similar effect was observed after muscimol-enhanced media (32.2 pg). Both agents also stimulated a substantial rise in intracellular calcium. Pretreatment of cultures with tetrodotoxin partially blacked the affects of muscimol on both peptide content and calcium activity, but not KCl. Calcium-depleted medium blacked the effects of KCl yet only attenuated the effects of muscimol. Treatment of cultures with cycloheximide blocked the effects of both muscimol and KCI. These results indicate that developing LHRH neurons are capable of synthesizing, secreting, and rapidly replenishing stores of LHRH peptide. C1 NINDS, Cellular & Dev Neurobiol Sect, NIH, Bethesda, MD 20895 USA. RP Wray, S (reprint author), NINDS, Cellular & Dev Neurobiol Sect, NIH, Bldg 36,Room 5A-25, Bethesda, MD 20895 USA. OI wray, susan/0000-0001-7670-3915 NR 33 TC 34 Z9 37 U1 0 U2 1 PU ENDOCRINE SOC PI BETHESDA PA 4350 EAST WEST HIGHWAY SUITE 500, BETHESDA, MD 20814-4110 USA SN 0013-7227 J9 ENDOCRINOLOGY JI Endocrinology PD DEC PY 2000 VL 141 IS 12 BP 4486 EP 4495 DI 10.1210/en.141.12.4486 PG 10 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 374TR UT WOS:000165360900021 PM 11108259 ER PT J AU Perfetti, R Zhou, J Doyle, ME Egan, JM AF Perfetti, R Zhou, J Doyle, ME Egan, JM TI Glucagon-like peptide-1 induces cell proliferation and pancreatic-duodenum homeobox-1 expression and increases endocrine cell mass in the pancreas of old, glucose-intolerant rats SO ENDOCRINOLOGY LA English DT Article ID TRANSCRIPTION FACTOR; DIABETES-MELLITUS; GENE; TOLERANCE; EXENDIN-4; MUTATIONS; DECLINE; PDX-1; MICE AB Glucose homeostasis in mammals is maintained by insulin secretion from the beta -cells of the islets of Langerhans. Type 2 diabetes results either from primary beta -cell failure alone and/or a failure to secrete enough insulin to overcome insulin resistance. Here, we show that continuous infusion of glucagon-like peptide-1 (7-36) (GLP-1; an insulinotropic agent), to young and old animals, had effects on the beta -cell of the pancreas other than simply on the insulin secretory apparatus. Our previous studies on a rodent model of glucose intolerance, the aging Wistar rat, show that a plateau in islet size, insulin content, and beta -cell mass is reached at 13 months, despite a continuing increase in body weight. Continuous sc infusion of GLP-1 (1.5 pM/kg.min), over 5 days, resulted in normal glucose tolerance. Our current results in both young and old rats demonstrate that treatment caused an up-regulation of pancreatic-duodenum homeobox-1 (PDX-1) expression in islets and total pancreas, induced pancreatic cell proliferation, and beta -cell neogenesis, The effects on levels of PDX-1 messenger RNA were abrogated by simultaneous infusion of Exendin (9-39), a specific antagonist of GLP-1. PDX-1 protein levels increased 4-fold in whole pancreata and 6-fold in islets in response to treatment. beta -cell mass increased to 7.2 +/- 0.58 from 4.88 +/- 0.38 mg, treated vs. control, respectively, P < 0.02. Total pancreatic insulin content also increased from 0.55 +/- 0.02 to 1.32 +/- 0.11 g/mg total pancreatic protein. Therefore, GLP-1 would seem to be a unique therapy that can stimulate pancreatic cell proliferation and beta -cell differentiation in the pancreas of rodents. C1 NIA, Ctr Gerontol Res, Diabet Sect, NIH, Baltimore, MD 21224 USA. Cedars Sinai Med Ctr, Div Endocrinol, Los Angeles, CA 90048 USA. RP Egan, JM (reprint author), NIA, Ctr Gerontol Res, Diabet Sect, NIH, Box 23,5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 22 TC 296 Z9 309 U1 0 U2 8 PU ENDOCRINE SOC PI BETHESDA PA 4350 EAST WEST HIGHWAY SUITE 500, BETHESDA, MD 20814-4110 USA SN 0013-7227 J9 ENDOCRINOLOGY JI Endocrinology PD DEC PY 2000 VL 141 IS 12 BP 4600 EP 4605 DI 10.1210/en.141.12.4600 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 374TR UT WOS:000165360900035 PM 11108273 ER PT J AU Ficca, SA Chyun, YD Ebrahimi, M Kutlak, F Memarzadeh, F AF Ficca, SA Chyun, YD Ebrahimi, M Kutlak, F Memarzadeh, F TI Activities of the National Institutes of Health relating to energy efficiency and pollution prevention SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article; Proceedings Paper CT Leadership Conference on Biomedical Research and the Environment CY NOV 01-02, 1999 CL BETHESDA, MARYLAND SP Natl Assoc Physicians Environm, Assoc Higher Educ Facil Officers, US Natl Inst Environm Hlth Sci, W Alton Jones Fdn, Mitchell Kapor Fdn, Shulsky Fdn, Hahn Family Fdn, Baxter Int Inc DE biomedical laboratory design; biomedical research; environmental stewardship; hospital design; laboratory energy use; research energy use; research facility construction; research support infrastructure AB The National Institutes of Health (NIH) is one of the world's premier biomedical research centers. Although NIH owns and operates more than 1,300 acres and 197 buildings across the country, the main campus is in Bethesda, Maryland. This campus consists of over 312 acres and 75 laboratories and other buildings, which consume vast amounts of energy. Aware of the NIH role in setting biomedical research agendas and priorities, its administrators strive to set good examples in energy efficiency and pollution prevention. Three current projects are presented as "best practices" examples of meeting the stated commitment of NIH to leadership in environmental stewardship: a) design and current construction of a 250-bed clinical research hospital designed to allow conversion of patient care units to research laboratories and vice-versa; b) design and construction of a six-story research laboratory that combines energy-saving innovations with breakthroughs in research technologies; and c) a massive, $200-million modernization of the campus utility infrastructure that involves generation systems for steam and chilled water and distribution systems for chilled water, steam, potable water, electricity, communications and computer networking, compressed air, and natural gas. Based on introduction of energy-efficiency measures, millions of dollars in savings for energy needs are projected; already the local electric utility has granted several million dollars in rebates. The guiding principles of NIH environmental stewardship help to ensure that energy conservation measures maximize benefits versus cost and also balance expediency with efficiency within available funding resources. This is a committee report for the Leadership Conference: Biomedical Research and the Environment held 1-2 November 1999 at the National Institutes of Health, Bethesda, Maryland. Key words: biomedical laboratory design, biomedical research, environmental stewardship, hospital design, laboratory energy use, research energy use, research facility construction, research support infrastructure. C1 NIH, Off Res Serv, Bethesda, MD 20892 USA. RP Ficca, SA (reprint author), NIH, Off Res Serv, 9000 Rockville Pike 21, Bethesda, MD 20892 USA. NR 1 TC 1 Z9 1 U1 0 U2 5 PU US DEPT HEALTH HUMAN SCIENCES PUBLIC HEALTH SCIENCE PI RES TRIANGLE PK PA NATL INST HEALTH, NATL INST ENVIRONMENTAL HEALTH SCIENCES, PO BOX 12233, RES TRIANGLE PK, NC 27709-2233 USA SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD DEC PY 2000 VL 108 SU 6 BP 939 EP 944 PG 6 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 386TP UT WOS:000166077900002 PM 11121359 ER PT J AU Barker, LF Rau, EH Pfister, EA Calcagni, J AF Barker, LF Rau, EH Pfister, EA Calcagni, J TI Development of a pollution prevention and energy efficiency clearinghouse for biomedical research facilities SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article; Proceedings Paper CT Leadership Conference on Biomedical Research and the Environment CY NOV 01-02, 1999 CL BETHESDA, MARYLAND SP Natl Assoc Physicians Environm, Assoc Higher Educ Facil Officers, US Natl Inst Environm Hlth Sci, W Alton Jones Fdn, Mitchell Kapor Fdn, Shulsky Fdn, Hahn Family Fdn, Baxter Int Inc DE biomedical research facilities; clearinghouse; energy efficiency; laboratory; pollution prevention AB This is the report of the National Association of Physicians for the Environment Committee on Development of a Pollution Prevention and Energy Efficiency Clearinghouse for Biomedical Research Facilities from the Leadership Conference on Biomedical Research and the Environment held at the National Institutes of Health in Bethesda. Maryland, on 1-2 November 1999. A major goal of the conference was the establishment of a World Wide Web-based clearinghouse, which would lend tremendous resources to the biomedical research community by providing accessto a database of peer-reviewed articles and references dealing with a host of aspects of biomedical research relating to energy efficiency, pollution prevention, and waste reduction. A temporary website has been established with the assistance of the U.S. Environmental Protection Agency (EPA) Regions ill and IV, where a pilot site provides access to the EPA's existing databases on these topics. A system of peer review for articles and promising techniques still must be developed, but a glimpse of topics and search engines is available for comment and review on the EPA Region IV-supported website (http://wrrc.p2pays.org/). C1 Sequelle Global TB Fdn, Rockville, MD 20850 USA. NIH, Off Res Serv, Div Safety, Environm Protect Branch, Bethesda, MD USA. US EPA, Waste Reduct Resource Ctr, Raleigh, NC USA. RP Barker, LF (reprint author), Sequelle Global TB Fdn, 9610 Med Ctr Dr,Suite 220, Rockville, MD 20850 USA. NR 2 TC 1 Z9 3 U1 0 U2 2 PU US DEPT HEALTH HUMAN SCIENCES PUBLIC HEALTH SCIENCE PI RES TRIANGLE PK PA NATL INST HEALTH, NATL INST ENVIRONMENTAL HEALTH SCIENCES, PO BOX 12233, RES TRIANGLE PK, NC 27709-2233 USA SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD DEC PY 2000 VL 108 SU 6 BP 949 EP 951 PG 3 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 386TP UT WOS:000166077900004 PM 11121361 ER PT J AU Rau, EH Alaimo, RJ Ashbrook, PC Austin, SM Borenstein, N Evans, MR French, HM Gilpin, RW Hughes, J Hummel, SJ Jacobsohn, AP Lee, CY Merkle, S Radzinski, T Sloane, R Wagner, KD Weaner, LE AF Rau, EH Alaimo, RJ Ashbrook, PC Austin, SM Borenstein, N Evans, MR French, HM Gilpin, RW Hughes, J Hummel, SJ Jacobsohn, AP Lee, CY Merkle, S Radzinski, T Sloane, R Wagner, KD Weaner, LE TI Minimization and management of wastes from biomedical research SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article; Proceedings Paper CT Leadership Conference on Biomedical Research and the Environment CY NOV 01-02, 1999 CL BETHESDA, MARYLAND SP Natl Assoc Physicians Environm, Assoc Higher Educ Facil Officers, US Natl Inst Environm Hlth Sci, W Alton Jones Fdn, Mitchell Kapor Fdn, Shulsky Fdn, Hahn Family Fdn, Baxter Int Inc DE biohazardous waste; biomedical research facilities; chemical waste; drug waste; hazardous waste; laboratories; medical waste; minimization; mixed waste; multihazardous waste; pollution prevention; radioactive waste; recycling; solid waste; training; waste disposal ID ISOLATED HEPATIC PERFUSION; TUMOR-NECROSIS-FACTOR; ANTIBIOTIC-RESISTANCE; MEDICAL WASTE; ESTROGENIC CHEMICALS; AQUATIC ENVIRONMENT; DOMESTIC SEWAGE; WATER SAMPLES; STW EFFLUENT; IDENTIFICATION AB Several committees were established by the National Association of Physicians for the Environment to investigate and report on various topics at the National Leadership Conference on Biomedical Research and the Environment held at the 1-2 November. 1999 at the National Institutes of Health in Bethesda, Maryland. This is the report of the Committee on Minimization and Management of Wastes from Biomedical Research. Biomedical research facilities contribute a small fraction of the total amount of wastes generated in the United States, and the rate of generation appears to be decreasing. Significant reductions in generation of hazardous, radioactive, and mixed wastes have recently been reported, even at facilities with rapidly expanding research programs. Changes in the focus of research, improvements in laboratory techniques, and greater emphasis on waste minimization (voiume and toxicity reduction) explain the declining trend in generation. The potential for uncontrolled releases of wastes from biomedical research facilities and adverse impacts on the general environment from these wastes appears to be low. Wastes are subject to numerous regulatory requirements and are contained and managed in a manner protective of the environment. Most biohazardous agents, chemicals, and radionuclides that find significant use in research are not likely to be persistent, bioaccumulative, or toxic if they are released. Today, the primary motivations for the ongoing efforts by facilities to improve minimization and management of wastes are regulatory compliance and avoidance of the high disposal costs and liabilities associated with generation of regulated wastes. The committee concluded that there was no evidence suggesting that the anticipated increases in biomedical research will significantly increase generation of hazardous wastes or have adverse impacts on the general environment. This conclusion assumes the positive. countervailing trends of enhanced pollution prevention efforts by facilities and reductions in waste generation resulting from improvements in research methods will continue. C1 NIH, Environm Protect Branch, Div Safety, Off Res Serv, Bethesda, MD 20892 USA. Procter & Gamble Pharmaceut, Norwich, NY USA. Univ Missouri, Columbia, MO USA. NIH, Radiat Safety Branch, Div Safety, Bethesda, MD 20892 USA. US EPA, Philadelphia, PA USA. NIH, Ctr Clin, Bethesda, MD 20892 USA. Univ Virginia, MERCI Program, Ctr Hlth Sci Operating Room, Charlottesville, VA USA. Johns Hopkins Univ, Baltimore, MD USA. Univ Maryland, Baltimore, MD 21201 USA. Natl Inst Environm Hlth Sci, Div Extramural Res & Training, Educ & Training Program, Res Triangle Pk, NC USA. Environm Ind Assoc, Washington, DC USA. Natl Inst Environm Hlth Sci, Off Management, Safety & Hlth Branch, Res Triangle Pk, NC USA. Natl Inst Environm Hlth Sci, Toxicol Lab, Div Intramural Res, Res Triangle Pk, NC USA. PRIZIM Inc, Gaithersburg, MD USA. RW Johnson Pharmaceut Res Inst, Spring House, PA 19477 USA. RP Rau, EH (reprint author), NIH, Environm Protect Branch, Div Safety, Off Res Serv, 13 South Dr,MSC 5746, Bethesda, MD 20892 USA. NR 182 TC 5 Z9 5 U1 2 U2 9 PU US DEPT HEALTH HUMAN SCIENCES PUBLIC HEALTH SCIENCE PI RES TRIANGLE PK PA NATL INST HEALTH, NATL INST ENVIRONMENTAL HEALTH SCIENCES, PO BOX 12233, RES TRIANGLE PK, NC 27709-2233 USA SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD DEC PY 2000 VL 108 SU 6 BP 953 EP 977 PG 25 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 386TP UT WOS:000166077900005 PM 11121362 ER PT J AU Wilson, SH Merkle, S Brown, D Moskowitz, J Hurley, D Brown, D Bailey, BJ McClain, M Misenhimer, M Buckalew, J Burks, T AF Wilson, SH Merkle, S Brown, D Moskowitz, J Hurley, D Brown, D Bailey, BJ McClain, M Misenhimer, M Buckalew, J Burks, T TI Biomedical research leaders: Report on needs, opportunities, difficulties, education and training, and evaluation SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article; Proceedings Paper CT Leadership Conference on Biomedical Research and the Environment CY NOV 01-02, 1999 CL BETHESDA, MARYLAND SP Natl Assoc Physicians Environm, Assoc Higher Educ Facil Officers, US Natl Inst Environm Hlth Sci, W Alton Jones Fdn, Mitchell Kapor Fdn, Shulsky Fdn, Hahn Family Fdn, Baxter Int Inc DE best practices; biomedical; database; environmental health; environmental stewardship; laboratory; research; sustainable development AB The National Association of Physicians for the Environment (NAPE) has assumed a leadership role in protecting environmental health in recent years. The Committee of Biomedical Research Leaders was convened at the recent NAPE Leadership Conference: Biomedical Research and the Environment held on 1-2 November 1999, at the National Institutes of Health, Bethesda, Maryland. This report summarizes the discussion of the committee and its recommendations The charge to the committee was to raise and address issues that will promote and sustain environmental health, safety, and energy efficiency within the biomedical community. Leaders from every important research sector (industry laboratories, academic health centers and institutes, hospitals and care facilities, Federal laboratories, and community-based research facilities) were gathered in this committee to discuss issues relevant to promoting environmental health. The conference and this report focus on the themes of environmental stewardship, sustainable development and "best greening practices." Environmental stewardship, an emerging theme within and outside the biomedical community, symbolizes the effort to provide an integrated, synthesized, and concerted effort to protect the health of the environment in both the present and the future. The primary goal established by the committee is to promote environmentally responsible leadership in the biomedical research community. Key outcomes of the committee's discussion and deliberation were a) the need for a central organization to evaluate, promote, and oversee efforts in environmental stewardship; and b) immediate need to facilitate efficient information transfer relevant to protecting the global environment through a database/clearinghouse. Means to fulfill these needs are discussed in this report. C1 NIEHS, Res Triangle Pk, NC 27709 USA. Wake Forest Univ, Winston Salem, NC 27109 USA. Univ Texas, Med Branch, Galveston, TX 77550 USA. Hoffmann La Roche Inc, Nutley, NJ 07110 USA. Univ So Calif, Los Angeles, CA USA. Powers Pyles Sutter & Verville, Washington, DC USA. Univ Texas, Hlth Sci Ctr, Houston, TX USA. RP Wilson, SH (reprint author), NIEHS, POB 12233, Res Triangle Pk, NC 27709 USA. NR 0 TC 3 Z9 3 U1 0 U2 1 PU US DEPT HEALTH HUMAN SCIENCES PUBLIC HEALTH SCIENCE PI RES TRIANGLE PK PA NATL INST HEALTH, NATL INST ENVIRONMENTAL HEALTH SCIENCES, PO BOX 12233, RES TRIANGLE PK, NC 27709-2233 USA SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD DEC PY 2000 VL 108 SU 6 BP 979 EP 995 PG 17 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 386TP UT WOS:000166077900006 PM 11121363 ER PT J AU Hodgson, M Brodt, W Henderson, D Loftness, V McCrone, R Roselle, G Rosenfeld, A Woods, J Wright, R AF Hodgson, M Brodt, W Henderson, D Loftness, V McCrone, R Roselle, G Rosenfeld, A Woods, J Wright, R TI Needs and opportunities for improving the health, safety, and productivity of medical research facilities SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article; Proceedings Paper CT Leadership Conference on Biomedical Research and the Environment CY NOV 01-02, 1999 CL BETHESDA, MARYLAND SP Natl Assoc Physicians Environm, Assoc Higher Educ Facil Officers, US Natl Inst Environm Hlth Sci, W Alton Jones Fdn, Mitchell Kapor Fdn, Shulsky Fdn, Hahn Family Fdn, Baxter Int Inc DE best practices; built environment; construction; emerging pathogens; healthy buildings; indoor environment; medical research; occupational health; preventive strategies; productivity; sick building; standards ID SICK-BUILDING SYNDROME; SYMPTOMS; TRANSMISSION; ASTHMA; OFFICE; RISK AB Medical research facilities, indeed all the nation's constructed facilities, must be designed, operated, and maintained in a manner that supports the health, safety, and productivity of the occupants. The National Construction Goals, established by the National Science and Technology Council, envision substantial improvements in occupant health and worker productivity. The existing research and best practices case studies support this conclusion, but too frequently building industry professionals lack the knowledge to design, construct, operate, and maintain facilities at these optimum levels. There is a need for more research and more collaborative efforts between medical and facilities engineering researchers and practitioners in order to attain the National Construction Goals. Such collaborative efforts will simultaneously support attainment of the National Health Goals. This article is the summary report of the Healthy Buildings Committee for the Leadership Conference: Biomedical Facilities and the Environment sponsored by the National institutes of Health, the National Association of Physicians for the Environment, and the Association of Higher Education Facilities Officers on 1-2 November 1999 in Bethesda, Maryland, USA. C1 NIH, Bethesda, MD 20892 USA. Dept Vet Affairs, Washington, DC USA. Carnegie Mellon Univ, Pittsburgh, PA 15213 USA. Dept Vet Affairs, Cincinnati, OH USA. Dept Energy, Washington, DC USA. HP Woods Res Inst, Herndon, VA USA. Natl Inst Stand & Technol, Gaithersburg, MD 20899 USA. RP Brodt, W (reprint author), NIH, Bldg 13,Rm 2W48, Bethesda, MD 20892 USA. NR 44 TC 1 Z9 1 U1 0 U2 0 PU US DEPT HEALTH HUMAN SCIENCES PUBLIC HEALTH SCIENCE PI RES TRIANGLE PK PA NATL INST HEALTH, NATL INST ENVIRONMENTAL HEALTH SCIENCES, PO BOX 12233, RES TRIANGLE PK, NC 27709-2233 USA SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD DEC PY 2000 VL 108 SU 6 BP 1003 EP 1008 PG 6 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 386TP UT WOS:000166077900008 PM 11124125 ER PT J AU Goldman, M Hedetniemi, JN Herbert, ER Sassaman, JS Walker, BC AF Goldman, M Hedetniemi, JN Herbert, ER Sassaman, JS Walker, BC TI Community outreach at biomedical research facilities SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article; Proceedings Paper CT Leadership Conference on Biomedical Research and the Environment CY NOV 01-02, 1999 CL BETHESDA, MARYLAND SP Natl Assoc Physicians Environm, Assoc Higher Educ Facil Officers, US Natl Inst Environm Hlth Sci, W Alton Jones Fdn, Mitchell Kapor Fdn, Shulsky Fdn, Hahn Family Fdn, Baxter Int Inc DE active and informed public outreach; community involvement; defusing public outrage; public/private partnerships AB For biomedical researchers to fulfill their responsibility for protecting the environment, they must do more than meet the scientific challenge of reducing the number and volume of hazardous materials used in their laboratories and the engineering challenge of reducing pollution and shifting to cleaner energy sources. They must also meet the public relations challenge of informing and involving their neighbors in these efforts. The experience of the Office of Community Liaison of the National Institutes of Health (NIH) in meeting the latter challenge offers a model and several valuable lessons for other biomedical research facilities to follow. This paper is based on presentations by an expert panel during the Leadership Conference on Biomedical Research and the Environment held 1-2 November 1999 at NIH, Bethesda, Maryland. The risks perceived by community members are often quite different from those identified by officials at the biomedical research facility. The best antidote for misconceptions is more and better information. If community organizations are to be informed participants in the decision-making process, they need a simple but robust mechanism for identifying and evaluating the environmental hazards in their community. Local government can and should be an active and fully informed partner in planning and emergency preparedness. In some cases this can reduce the regulatory burden on the biomedical research facility. In other cases it might simplify and expedite the permitting process or help the facility disseminate reliable information to the community. When a particular risk, real or perceived, is of special concern to the community, community members should be involved in the design, implementation, and evaluation of targeted risk assessment activities. Only by doing so will the community have confidence in the results of those activities. NIH has involved community members in joint efforts to deal with topics as varied as recycling and soil testing. These ad hoc efforts are more likely to succeed if community members and groups have also been included in larger and longer term advisory committees. These committees institutionalize the outreach process. This can provide the facility with vocal and influential allies who create an independent line of communication with the larger community. C1 NIH, Off Community Liaison, Bethesda, MD 20892 USA. Dept Environm Protect, Rockville, MD USA. Opus Grp LLC, Chapel Hill, NC USA. Howard Univ, Med Ctr, Off Environm & Occupat Med, Washington, DC 20059 USA. RP Hedetniemi, JN (reprint author), NIH, Off Community Liaison, Bldg 1,Rm 259, Bethesda, MD 20892 USA. NR 1 TC 0 Z9 0 U1 1 U2 1 PU US DEPT HEALTH HUMAN SCIENCES PUBLIC HEALTH SCIENCE PI RES TRIANGLE PK PA NATL INST HEALTH, NATL INST ENVIRONMENTAL HEALTH SCIENCES, PO BOX 12233, RES TRIANGLE PK, NC 27709-2233 USA SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD DEC PY 2000 VL 108 SU 6 BP 1009 EP 1013 PG 5 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 386TP UT WOS:000166077900009 PM 11124126 ER PT J AU Dong, WM Kari, FW Selgrade, MK Gilmour, MI AF Dong, WM Kari, FW Selgrade, MK Gilmour, MI TI Attenuated allergic responses to house dust mite antigen in feed-restricted rats SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article DE allergy; asthma; dust mites; eosinophil; feed restriction; IgE; immune response; inflammation; lung; T lymphocyte; tumor necrosis factor-alpha ID TUMOR-NECROSIS-FACTOR; COLONY-STIMULATING-FACTOR; TNF-ALPHA; FOOD RESTRICTION; CALORIE RESTRICTION; EPITHELIAL-CELLS; GUINEA-PIG; INFLAMMATION; EXPRESSION; GLUCOCORTICOIDS AB Caloric restriction has been shown to alter a broad range of immunological end points in both experimental animals and humans. The objective of this study was to investigate the effect of short-term moderate feed restriction (25% reduction) on allergic immune responses in Brown Norway rats. After 3 weeks of acclimation to their feed regimens, rats were sensitized and 2 weeks later challenged with house dust mite (HDM) antigen via intratracheal instillation. Feed restriction resulted in lower levels of antigen-specific IgE in serum and reduced antigenspecific lymphoproliferative activity in pulmonary lymph nodes. Feed restriction also attenuated pulmonary inflammation, as evidenced by lower levels of lactate dehydrogenase and total protein, decreased infiltration of neutrophils and eosinophils, and reduced secretion of pro-inflammatory cytokine tumor necrosis factor (TNF)-alpha in bronchoalveolar lavage fluid. In addition, feed restriction decreased TNF-alpha secretion in serum and decreased mRNA expression of TNF-alpha and interleukin-6 in pulmonary lymph nodes. We conclude that feed restriction strongly dampened the allergic immune responses to HDM in rats and that this attenuation was associated with decreased expression and secretion of pro-inflammatory cytokines. C1 NIEHS, Lab Environm Carcinogenesis & Mutagenesis, Res Triangle Pk, NC 27709 USA. US EPA, Immunotoxicol Branch, Res Triangle Pk, NC 27711 USA. RP Kari, FW (reprint author), NIEHS, Lab Environm Carcinogenesis & Mutagenesis, MD F1-05,POB 12233,111 TW Alexander Dr, Res Triangle Pk, NC 27709 USA. NR 32 TC 12 Z9 12 U1 1 U2 2 PU US DEPT HEALTH HUMAN SCIENCES PUBLIC HEALTH SCIENCE PI RES TRIANGLE PK PA NATL INST HEALTH, NATL INST ENVIRONMENTAL HEALTH SCIENCES, PO BOX 12233, RES TRIANGLE PK, NC 27709-2233 USA SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD DEC PY 2000 VL 108 IS 12 BP 1125 EP 1131 DI 10.2307/3434823 PG 7 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 385LJ UT WOS:000166005700022 PM 11133391 ER PT J AU Gomez, E Peguero, M Sanchez, J Castellanos, PL Feris, J Pena, C Brudzinski-LaClaire, L Levine, OS AF Gomez, E Peguero, M Sanchez, J Castellanos, PL Feris, J Pena, C Brudzinski-LaClaire, L Levine, OS TI Population-based surveillance for bacterial meningitis in the Dominican Republic: implications for control by vaccination SO EPIDEMIOLOGY AND INFECTION LA English DT Article ID HAEMOPHILUS-INFLUENZAE; YOUNG-CHILDREN; DISEASE BURDEN; VACCINES; IMPLEMENTATION; PREVENTION; FRAMEWORK; COUNTRIES; INFANTS; CHILE AB Quantifying the local burden of disease is an important step towards the introduction of new vaccines, such as Haemophilus influenzae type b (Hib) conjugate vaccine. We adapted a generic protocol developed by the World Health Organization for population-based surveillance of bacterial meningitis. All hospitals that admit paediatric patients with meningitis in the National District, Dominican Republic were included in the system and standard laboratory methods were used. The system identified 111 cases of confirmed bacterial meningitis. Hib was the leading cause of bacterial meningitis, followed by group B streptococcus, S. pneumoniae, and N. meningitidis. Unlike hospital-based case series, this population-based system was able to calculate incidence rates. The incidence of Hib meningitis was 13 cases per 100 000 children < 5 years old. The data from this study were used by the Ministry of Health to support the introduction of routine Hib vaccination and will be used to monitor its effectiveness. C1 Secretaria Estado Salud Publ & Asistencia Social, Santo Domingo, Dominican Rep. Clin Infantil Dr Robert Reid Cabral, Dept Infectol, Santo Domingo, Dominican Rep. Pan Amer Hlth Org, Santo Domingo, Dominican Rep. Ctr Dis Control & Prevent, Resp Dis Branch, Div Bacterial & Mycot Dis, Natl Ctr Infect Dis, Atlanta, GA USA. RP Levine, OS (reprint author), NIAID, DMID, RDB, 6700-B Rockledge Dr,Room 3255,MSC 7630, Bethesda, MD 20892 USA. NR 20 TC 6 Z9 7 U1 0 U2 2 PU CAMBRIDGE UNIV PRESS PI PORT CHESTER PA 110 MIDLAND AVE, PORT CHESTER, NY 10573-4930 USA SN 0950-2688 J9 EPIDEMIOL INFECT JI Epidemiol. Infect. PD DEC PY 2000 VL 125 IS 3 BP 549 EP 554 DI 10.1017/S0950268800004830 PG 6 WC Public, Environmental & Occupational Health; Infectious Diseases SC Public, Environmental & Occupational Health; Infectious Diseases GA 422BF UT WOS:000168097500010 PM 11218205 ER PT J AU Lo, SC Levin, L Ribas, J Chung, R Wang, RYH Wear, D Shih, JWK AF Lo, SC Levin, L Ribas, J Chung, R Wang, RYH Wear, D Shih, JWK TI Lack of serological evidence for Mycoplasma fermentans infection in army Gulf War veterans: a large scale case-control study SO EPIDEMIOLOGY AND INFECTION LA English DT Article ID AIDS; ASSOCIATION; INCOGNITUS; PENETRANS; HIV AB Mycoplasma fermentans is suspected in the development of 'Gulf War illness' in veterans of Operation Desert Storm. We conducted a matched case-control study for the prevalence of M. fermentans-specific antibodies before and after the operation, as well as seroconversion rates in veterans with and without complaints of 'Gulf War illness'. Cases consisted of Gulf War veterans, who complained of Various illnesses and were enrolled in the second phase of the health evaluation by the Army Comprehensive Clinical Examination Program (CCEP). Controls were selected from Gulf War veterans who did not participate in the registry and did not request a health evaluation by the CCEP. Before operation deployment, 34 out of 718 of the cases (4.8%) and 116 out of 2233 of the controls (5.2%) tested positive for M. fermentans-specific antibodies. There was no difference in rates of seroconversion between cases and controls (1.1 vs. 1.2%) to M. fermentans during Operation Desert Storm. Thus, there is no serological evidence that suggests infection by M, fermentans is associated with development of 'Gulf War illness'. C1 Armed Forces Inst Pathol, Dept Infect & Parasit Dis Pathol, Washington, DC 20306 USA. Walter Reed Army Med Ctr, Walter Reed Army Inst Res, Dept Epidemiol, Div Prevent Med, Washington, DC 20307 USA. Walter Reed Army Med Ctr, Infect Dis Serv, Washington, DC 20307 USA. NIH, Warren Grant Magnuson Clin Ctr, Dept Transfus Med, Bethesda, MD 20892 USA. RP Lo, SC (reprint author), Armed Forces Inst Pathol, Dept Infect & Parasit Dis Pathol, 14th St & Alaska Ave NW, Washington, DC 20306 USA. NR 25 TC 5 Z9 5 U1 0 U2 0 PU CAMBRIDGE UNIV PRESS PI PORT CHESTER PA 110 MIDLAND AVE, PORT CHESTER, NY 10573-4930 USA SN 0950-2688 J9 EPIDEMIOL INFECT JI Epidemiol. Infect. PD DEC PY 2000 VL 125 IS 3 BP 609 EP 616 DI 10.1017/S0950268800004891 PG 8 WC Public, Environmental & Occupational Health; Infectious Diseases SC Public, Environmental & Occupational Health; Infectious Diseases GA 422BF UT WOS:000168097500017 PM 11218212 ER PT J AU Postma, T Krupp, E Li, XL Post, RM Weiss, SRB AF Postma, T Krupp, E Li, XL Post, RM Weiss, SRB TI Lamotrigine treatment during amygdala-kindled seizure development fails to inhibit seizures and diminishes subsequent anticonvulsant efficacy SO EPILEPSIA LA English DT Article DE epilepsy; kindling; amygdala; lamotrigine; rats ID GLUTAMATE-RECEPTOR ANTAGONISTS; PIMOZIDE BLOCKS ESTABLISHMENT; FIBER SYNAPTIC REORGANIZATION; LONG-TERM POTENTIATION; ANTIEPILEPTIC DRUGS; PHARMACOLOGICAL CHARACTERIZATION; CLINICAL-EXPERIENCE; STATUS EPILEPTICUS; BIPOLAR DISORDER; CALCIUM CURRENTS AB Purpose: Lamotrigine (LTG) is an anticonvulsant that is currently in use for the treatment of various seizure disorders and that shows promise in the treatment of affective illness. LTG is also effective in the suppression of amygdala-kindled seizures. Because many drugs show a differential efficacy profile as a function of the phase of kindling evolution, we evaluated LTG for its potential antiepileptogenic effects on the development of amygdala-kindled seizures. Methods: In two separate studies, LTG (5 or 15 mg/kg versus vehicle) was administered before each daily amygdala stimulation (biphasic square wave pulses, 100 pulse pairs per second for a total of 0.5 second, I-millisecond pulse width) at an intensity of 50 muA over the AD threshold. Seizure development was assessed, as well as the effect of this pretreatment on subsequent efficacy of LTG on completed kindled seizures. Results: LTG at 5 mg/kg failed to block seizure development. At 15 mg/kg, LTG paradoxically enhanced seizure development and produced running fits in four of the nine animals tested. Animals previously treated with either dose of LTG during kindling development showed a diminished response to the anticonvulsant effects of LTG on fully kindled seizures compared with the vehicle-treated controls. Conclusions: Although LTG possesses potent anticonvulsant effects on completed amygdala-kindled seizures, it is either without effect (5 mg/kg) or facilitates (15 mg/kg) the initial phase of kindling development. In addition, exposure to LTG during kindled seizure development leads to a reduced subsequent response to the drug in fully kindled animals. These observations parallel those with carbamazepine and suggest that different stages of kindling (epileptogenesis versus fully manifest seizures) may have different underlying neural mechanisms that require distinct pharmacotherapies. C1 NIMH, BPB, Bethesda, MD 20892 USA. RP Weiss, SRB (reprint author), NIMH, BPB, Bldg 10-3S239,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 74 TC 49 Z9 49 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0013-9580 J9 EPILEPSIA JI Epilepsia PD DEC PY 2000 VL 41 IS 12 BP 1514 EP 1521 PG 8 WC Clinical Neurology SC Neurosciences & Neurology GA 382RG UT WOS:000165835600003 PM 11114208 ER PT J AU Li, B Cao, DJ Xu, HW Chang, JL Zhou, GS Tian, JX Li, DJ Theze, J Wu, CY AF Li, B Cao, DJ Xu, HW Chang, JL Zhou, GS Tian, JX Li, DJ Theze, J Wu, CY TI Interleukin-12 induces genes expression in interleukin-2 stimulated human T lymphocytes SO EUROPEAN CYTOKINE NETWORK LA English DT Article DE IL-12; IL-2; T lymphocytes; C-type lectin; glucose transporter-like protein ID HELPER CELL-DIFFERENTIATION; GLUT3 EXPRESSION; IL-12 RECEPTOR; IFN-GAMMA; HUMAN NK; ACTIVATION; PROTEIN; PHOSPHORYLATION; TRANSPORT; COMPLEX AB IL-12 is a critical immunoregulatory cytokine that promotes cell-mediated immune responses by inducing the differentiation of Th1 cells. To better clarify the molecular basis of IL-12 action, we compared the gene expression in human T lymphocytes activated by IL-2 and IL-12. mRNAs from T lymphocytes activated by either IL-2 alone or IL-2 plus IL-12 were transcribed into cDNAs. A differential mRNA display was conducted. As a result, differential display of five cDNA fragments was obtained. Sequence analysis suggests that they had high homology with recorded genes as found by a computer search against GenBank. Two full genes of the five fragments were cloned, which activation-induced C-type lectin and glucose transporter-like protein. Interestingly, these proteins were expressed in the T cells stimulated by IL-2 and IL-12, but not in the T cells stimulated by IL-2 alone. These results suggest that C-type lectin and glucose transporter-like protein may play an important role in the T lymphocyte activation induced by IL-12. C1 Harbin Med Univ, Dept Immunol, Harbin 150086, Peoples R China. Natl Key Lab Vet Biotechnol, Harbin 150001, Peoples R China. Inst Pasteur, Div Cell Immunol & Immunogen, Paris, France. NIH, Natl Inst Allergy & Infect Dis, Lab Clin Invest, Bethesda, MD USA. RP Li, B (reprint author), Harbin Med Univ, Dept Immunol, Harbin 150086, Peoples R China. NR 37 TC 3 Z9 3 U1 0 U2 0 PU JOHN LIBBEY EUROTEXT LTD PI MONTROUGE PA 127 AVE DE LA REPUBLIQUE, 92120 MONTROUGE, FRANCE SN 1148-5493 J9 EUR CYTOKINE NETW JI Eur. Cytokine Netw. PD DEC PY 2000 VL 11 IS 4 BP 602 EP 607 PG 6 WC Biochemistry & Molecular Biology; Cell Biology; Immunology SC Biochemistry & Molecular Biology; Cell Biology; Immunology GA 393NW UT WOS:000166477400021 PM 11125303 ER PT J AU Marx, C Bornstein, SR Wolkersdorfer, GW AF Marx, C Bornstein, SR Wolkersdorfer, GW TI Cellular immune-endocrine interaction in adrenocortical tissues SO EUROPEAN JOURNAL OF CLINICAL INVESTIGATION LA English DT Article; Proceedings Paper CT Conference on Adrenal 2000 - From Molecular Basics to New Clinical Dimensions CY 2000 CL HAMBURG, GERMANY DE adrenal gland; androgen production; immune-endocrine interaction ID CLASS-II EXPRESSION; RELEVANCE; APOPTOSIS AB Background The detection of important immunocompetence-like features on endocrine steroid cells raises questions about direct intercellular communication between the adrenal and immune systems. This article summarizes our recent work and new data on immune-adrenal interactions. Materials and methods RT-PCR and immunohistochemistry were performed to examine MHC class II (HLA-DR) expression in adrenocortical rumours. Coculture systems of NCI-H295 adrenocortical carcinoma cells and HLA-marched lymphocytes were used to examine effects on steroid production and survival of lymphocytes. Results HLA-DR m-RNA is found in both benign and malignant adrenals, except the NCI-H295 cell line. Under direct coculture conditions with NCI-H295 cell line, spontaneous apoptosis of immune cells was reduced. Synthesis of cortisol and especially of dehydroepiandrosterone production of tumour cells was markedly increased. Differences by separating CD++ and CD8+ T cells were not detected. Conclusions Direct cellular contact between lymphocytes and adrenocortical cells seems to be involved in the peripheral regulation of androgen synthesis in the adrenal. The molecular basis of this interaction is no tknown. With regard to normal adrenals, ligation of MHC class II antigens could be a potential mechanism for a peripheral regulation of androgen secretion. C1 NICHHD, NIH, Ctr Clin, Bethesda, MD 20814 USA. Univ Leipzig, Dept Internal Med 3, Leipzig, Germany. RP Bornstein, SR (reprint author), NICHHD, NIH, Ctr Clin, Bldg 10,Room 10N262, Bethesda, MD 20814 USA. NR 12 TC 8 Z9 10 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0014-2972 J9 EUR J CLIN INVEST JI Eur. J. Clin. Invest. PD DEC PY 2000 VL 30 SU 3 BP 1 EP 5 DI 10.1046/j.1365-2362.2000.0300s3001.x PG 5 WC Medicine, General & Internal; Medicine, Research & Experimental SC General & Internal Medicine; Research & Experimental Medicine GA 405RD UT WOS:000167172200002 PM 11281360 ER PT J AU Glasow, A Bornstein, SR AF Glasow, A Bornstein, SR TI Leptin and the adrenal gland SO EUROPEAN JOURNAL OF CLINICAL INVESTIGATION LA English DT Article; Proceedings Paper CT Conference on Adrenal 2000 - From Molecular Basics to New Clinical Dimensions CY 2000 CL HAMBURG, GERMANY DE adrenal gland; leptin ID LASER CAPTURE MICRODISSECTION; ACTIVATED PROTEIN-KINASE; IN-SITU HYBRIDIZATION; PANCREATIC BETA-CELLS; OBESE GENE-EXPRESSION; ADIPOSE-TISSUE; PLASMA LEPTIN; INSULIN-RESISTANCE; CUSHINGS-SYNDROME; MESSENGER-RNA AB Background Leptin is involved in the maintenance of energy balance acting on food intake, thermogenesis and energy expenditure. Via its receptor in the hypothalamus, leptin modulates the functioning of the hypothalamic-pituitary-adrenal axis and the systemic sympathetic/adrenomedullary system, which are closely linked to the regulation of energy balance and body weight. In regard of potential interactions of leptin and adrenal hormones this study intended to characterize the role of leptin in the human adrenal gland. Materials and methods A novel technique of laser capture microdissection was used to separate cortical and chromaffin cells for mRNA expression studies of leptin receptor isoforms and leptin mRNA in adrenal tissue and cell line NCI-H295. Immunostaining was used to localize leptin receptor in human adrenal slices. The influence of leptin on basal and ACTH-stimulated steroid hormone secretion and enzyme expression was assessed. The effect of leptin on proliferation and viability of adrenal cells in primary culture and of the NCI-H295 cell line was studied by the WST-1 assay and by H-1-thymidine test. Results Our data demonstrate that leptin can regulate the human adrenal function directly, via its receptors on adrenocortical cells. Leptin decreased the corticotropin-stimulated release of steroid hormones In vitro without any effect on cell proliferation. No influence of leptin on the expression of cytochrome P450(17 alpha) and P450(SCC) mRNA was detected. Conclusions The adipo-adrenal interaction mediated by leptin further underscores the close link of metabolism and stress regulation in humans. C1 Univ Leipzig, Dept Internal Med 3, D-7010 Leipzig, Germany. NICHHD, NIH, Bethesda, MD 20892 USA. RP Glasow, A (reprint author), Inst Canc Res, Chester Beatty Labs, Leukaemia Res Fund Ctr, 237 Fulham Rd, London SW3 6JB, England. NR 93 TC 41 Z9 43 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0014-2972 J9 EUR J CLIN INVEST JI Eur. J. Clin. Invest. PD DEC PY 2000 VL 30 SU 3 BP 39 EP 45 DI 10.1046/j.1365-2362.2000.0300s3039.x PG 7 WC Medicine, General & Internal; Medicine, Research & Experimental SC General & Internal Medicine; Research & Experimental Medicine GA 405RD UT WOS:000167172200009 PM 11281366 ER PT J AU Path, G Scherbaum, WA Bornstein, SR AF Path, G Scherbaum, WA Bornstein, SR TI The role of interleukin-1 in the human adrenal gland SO EUROPEAN JOURNAL OF CLINICAL INVESTIGATION LA English DT Article; Proceedings Paper CT Conference on Adrenal 2000 - From Molecular Basics to New Clinical Dimensions CY 2000 CL HAMBURG, GERMANY DE adrenocortical steroids; HPA axis; IL-6 receptor; IL-6; stress ID MIGRATION-INHIBITORY FACTOR; PITUITARY-CELLS-INVITRO; TUMOR NECROSIS FACTOR; RECOMBINANT INTERLEUKIN-6; SIGNAL TRANSDUCER; ADRENOCORTICAL-CELLS; ANTITUMOR-ACTIVITY; ENDOCRINE SYSTEMS; RELEASE; RECEPTOR AB Interleukin (IL)-6 is a potent activator of the hypothalamic-pituitary-adrenal (HPA) axis on all levels in humans, and appears to play a pathogenic role in conditions related to chronic stress and physiological ageing; with physiological ageing showing a similar hormonal and immunological pattern to chronic stress. IL-6 and its receptor IL-6R are coexpressed at similar sites in the human adrenal gland, which seems to be an important source of IL-6 production. In vitro, in primary cultures of adrenal gland cells, chronic exposure to IL-6 stimulates adrenocortical steroid release in a time- and dose-dependent manner. This explains the high systemic cortisol levels in the absence of adequate plasma concentrations of corticotropin (ACTH) observed in patients after long-term treatment with IL-6. It could therefore be concluded that in situations of prolonged stress, when corticotropin-releasing hormone and ACTH release are suppressed by feedback inhibition due to circulating glucocorticoids, IL-6 maintains the elevated glucocorticoid levels by direct stimulation of adrenocortical steroidogenesis via autocrine/paracrine mechanisms. C1 Univ Dusseldorf, Diabet Forschungsinst, D-4000 Dusseldorf, Germany. NIH, Bethesda, MD 20892 USA. RP Path, G (reprint author), Univ Hosp, Baldingerstr, D-35033 Marburg, Germany. NR 64 TC 39 Z9 44 U1 0 U2 1 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0014-2972 J9 EUR J CLIN INVEST JI Eur. J. Clin. Invest. PD DEC PY 2000 VL 30 SU 3 BP 91 EP 95 DI 10.1046/j.1365-2362.2000.0300s3091.x PG 5 WC Medicine, General & Internal; Medicine, Research & Experimental SC General & Internal Medicine; Research & Experimental Medicine GA 405RD UT WOS:000167172200019 PM 11281377 ER PT J AU Semba, RD Kumwenda, N Hoover, DR Taha, TE Mtimavalye, L Broadhead, R Eisinger, W Miotti, PG Chiphangwi, JD AF Semba, RD Kumwenda, N Hoover, DR Taha, TE Mtimavalye, L Broadhead, R Eisinger, W Miotti, PG Chiphangwi, JD TI Assessment of iron status using plasma transferrin receptor in pregnant women with and without human immunodeficiency virus infection in Malawi SO EUROPEAN JOURNAL OF CLINICAL NUTRITION LA English DT Article DE pregnancy; iron; iron deficiency anemia; transferrin receptor; ferritin; hemoglobin; human immunodeficiency virus; Malawi ID DEFICIENCY ANEMIA; CHRONIC DISEASE; SERUM FERRITIN; PATHOGENESIS; UTILITY AB Background: Although anemia is highly prevalent during pregnancy and is common during human immunodeficiency virus (HIV) infection, anemia and iron status have not been well characterized in HIV-infected pregnant women. Objective: To gain insight into iron status in HIV-infected pregnant women using plasma transferrin receptor and related indicators of anemia. Study design: Plasma transferrin receptor, ferritin, alpha (1)-acid glycoprotein, C-reactive protein and hemoglobin concentrations were measured in pregnant women, gestational age 18-28 weeks, seen in an urban antenatal clinic in Blantyre, Malawi. Results: The prevalence of anemia among 662 HIV-positive and 190 HIV-negative pregnant women was 73.1% and 50.0%, respectively (P < 0.0001). Among HIV-positive and HIV-negative women, median plasma transferrin receptor concentrations were 24.4 and 24.1 nmol/l (P = 0.5), respectively, and median plasma ferritin concentrations were 17.8 and 20.8 g/l (P < 0.05), respectively. There was a large overlap in plasma transferrin receptor concentrations among women with and without anemia. Using the combination of hemoglobin and ferritin as a standard, the sensitivity and specificity of plasma transferrin receptor in diagnosing iron deficiency anemia was estimated at 45.9% and 68.1%, respectively. Conclusion: The use of plasma transferrin receptor concentrations as an indicator of iron deficiency anemia may be limited in pregnant women with chronic inflammation and infection. C1 Johns Hopkins Univ, Sch Med, Dept Ophthalmol, Baltimore, MD USA. Johns Hopkins Univ, Sch Med, Dept Epidemiol, Baltimore, MD USA. Johns Hopkins Univ, Sch Hyg & Publ Hlth, Dept Epidemiol, Baltimore, MD USA. Johns Hopkins Univ, Sch Hyg & Publ Hlth, Dept Ophthalmol, Baltimore, MD USA. Rutgers State Univ, Dept Stat, Piscataway, NJ 08855 USA. Univ Malawi, Coll Med, Dept Obstet & Gynaecol, Blantyre, Malawi. Univ Malawi, Coll Med, Dept Paediat & Child Hlth, Blantyre, Malawi. NIAID, Bethesda, MD 20892 USA. RP Semba, RD (reprint author), 550 N Broadway,Suite 700, Baltimore, MD 21205 USA. FU NIAID NIH HHS [N01-AI-35173-117]; NICHD NIH HHS [HD32247, HD30042] NR 28 TC 19 Z9 19 U1 0 U2 2 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0954-3007 J9 EUR J CLIN NUTR JI Eur. J. Clin. Nutr. PD DEC PY 2000 VL 54 IS 12 BP 872 EP 877 DI 10.1038/sj.ejcn.1601106 PG 6 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 384VA UT WOS:000165965600003 PM 11114684 ER PT J AU Egan, CA Yancey, KB AF Egan, CA Yancey, KB TI The clinical and immunopathological manifestations of anti-epiligrin cicatricial pemphigoid, a recently defined subepithelial autoimmune blistering disease SO EUROPEAN JOURNAL OF DERMATOLOGY LA English DT Review DE autoimmunity; epidermal basement membrane; extracellular matrix; IgG; laminin ID EPIDERMOLYSIS-BULLOSA ACQUISITA; BASEMENT-MEMBRANE; EXTRACELLULAR DOMAIN; AUTOANTIBODIES REACT; ANCHORING FILAMENTS; GASTRIC-CARCINOMA; VII COLLAGEN; HUMAN SKIN; LAMININ-5; BP180 AB Cicatricial pemphigoid (CP) is a rare, acquired, autoimmune. subepithelial blistering disease. It primarily affects mucous membranes but it also may involve the skin. Morbidity is associated with the propensity for scar formation and may be especially severe when mucosal surfaces such as the conjunctivae, larynx, esophagus, or urethra are involved. On direct immunofluorescence microscopy, CP is characterized by the linear deposition of immunoreactants, principally IgG and C3, along epithelial basement membranes. Over the last 10 years, studies in a number of laboratories have shown that circulating autoantibodies in patients with CP may target one of several different autoantigens. One subset of patients with the CP-phenotype have IgG anti-basement membrane autoantibodies against laminin 5 (alpha3 beta3 gamma2) (i.e., patients with anti-epiligrin CP [AECP]). This review discusses recent advances in the understanding of CP and emphasizes salient features of AECP pathophysiology. C1 NCI, Dermatol Branch, Div Clin Sci, NIH, Bethesda, MD 20892 USA. RP Egan, CA (reprint author), NCI, Dermatol Branch, Div Clin Sci, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 58 TC 24 Z9 24 U1 0 U2 0 PU JOHN LIBBEY EUROTEXT LTD PI MONTROUGE PA 127 AVE DE LA REPUBLIQUE, 92120 MONTROUGE, FRANCE SN 1167-1122 J9 EUR J DERMATOL JI Eur. J. Dermatol. PD DEC PY 2000 VL 10 IS 8 BP 585 EP 589 PG 5 WC Dermatology SC Dermatology GA 387ZX UT WOS:000166153900001 PM 11125317 ER PT J AU Marth, T Ring, S Schulte, D Klensch, N Strober, W Kelsall, BL Stallmach, A Zeitz, M AF Marth, T Ring, S Schulte, D Klensch, N Strober, W Kelsall, BL Stallmach, A Zeitz, M TI Antigen-induced mucosal T cell activation is followed by Th1 T cell suppression in continuously fed ovalbumin TCR-transgenic mice SO EUROPEAN JOURNAL OF IMMUNOLOGY LA English DT Article DE oral tolerance; IL-12; IL-10; IFN-gamma; Peyer's patch ID EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS; MYELIN BASIC-PROTEIN; DOSE ORAL TOLERANCE; IFN-GAMMA; CHOLERA-TOXIN; INDUCTION; MECHANISMS; RESPONSES; IMMUNIZATION; APOPTOSIS AB We investigated kinetics and dose-dependent features of mucosal and peripheral immune responses following oral antigen application in a TCR-transgenic mouse model. Ovalbumin (OVA) TCR-transgenic mice were fed OVA at different doses (5-250 mg) and various frequencies tone to seven times, or continuous feeding). Low- and medium-dose (10, 100 mg) OVA feeding resulted in priming of immune responses, i.e. increased antigen-specific proliferation as well as IL-2, IL-4 and IFN-1 secretion upon in vitro restimulation in Peyer's patches and spleen. Immune responses were suppressed with doses of one or three times 250 mg OVA feeding in the spleen. However, only the highest OVA feeding doses (7x250 mg OVA) or continuous feeding (5 mg daily in the drinking water over a 12-week period) actively suppressed immune responses and were associated with production of TGF-beta and IL-10 in the spleen and Peyer's patches. Thus, the cell population generated by continuous antigen feeding was characterized by production of suppressive cytokines and seems to be based on a counter-regulation with Th1 cytokines. These data further define the regulation of suppressive immune functions following antigen feeding in the periphery and the mucosal immune system. C1 Univ Saarlandes Kliniken, D-66421 Homburg, Germany. NIH, Mucosal Immun Sect, Bethesda, MD 20892 USA. RP Marth, T (reprint author), Univ Saarlandes Kliniken, D-66421 Homburg, Germany. NR 36 TC 32 Z9 34 U1 0 U2 0 PU WILEY-V C H VERLAG GMBH PI BERLIN PA PO BOX 10 11 61, D-69451 BERLIN, GERMANY SN 0014-2980 J9 EUR J IMMUNOL JI Eur. J. Immunol. PD DEC PY 2000 VL 30 IS 12 BP 3478 EP 3486 DI 10.1002/1521-4141(2000012)30:12<3478::AID-IMMU3478>3.0.CO;2-A PG 9 WC Immunology SC Immunology GA 384YU UT WOS:000165975500017 PM 11093167 ER PT J AU von Stebut, E Belkaid, Y Nguyen, BV Cushing, M Sacks, DL Udey, MC AF von Stebut, E Belkaid, Y Nguyen, BV Cushing, M Sacks, DL Udey, MC TI Leishmania major-infected murine Langerhans cell-like dendritic cells from susceptible mice release IL-12 after infection and vaccinate against experimental cutaneous Leishmaniasis SO EUROPEAN JOURNAL OF IMMUNOLOGY LA English DT Article DE dendritic cell; Langerhans cell; Leishmania major; IL-12; Th1 ID CD4(+) T-CELLS; TGF-BETA; IMMUNE-RESPONSE; BALB/C MICE; IFN-GAMMA; TH2 CELLS; EXPRESSION; MACROPHAGES; INTERLEUKIN-12; INDUCTION AB Leishmania major-infected C57BL/6 skin-dendritic cells (DC) are activated and release cytokines (including IL-12 p70), and likely initiate protective Th1 immunity in vivo (von Stebut, E. et al., J. Exp. Med. 188: 1547-1552). To characterize differences in DC function in mice that are genetically susceptible (BALB/c) and resistant (C57BL/6) to cutaneous leishmaniasis, eve analyzed the effects of L. major on Langerhans cell-like, fetal skin-derived DC (FSDDC) from both strains. BALB/c- and C57BL/6-FSDDC ingested similar numbers of amastigotes, but did not ingest metacyclic promastigotes. Like C57BL/6-FSDDC, infection of BALB/c-FSDDC led to up-regulation of MHC class I and II antigens, CD40, CD54, and CD86 within 18 h. L. major-induced BALB/c DC activation also led to the release of TNF-alpha, IL-6 and IL-12 p40 into 18-h supernatants. Infected BALB/c- and C57BL/6-DC both released small amounts of IL-12 p70 within 72 h. Additional stimulation with IFN-gamma and/or anti-CD40 induced the release of more IL-12 p70 from infected BALB/c-DC than C57BL/6-DC. Coculture of control or infected BALB/c- and C57BL/6-DC with naive syngeneic CD4(+) T cells and soluble anti-CD3 resulted in mixed, IFN-gamma -predominant responses after restimulation with immobilized anti-CDS. Finally syngeneic L. major-infected DC effectively vaccinated BALB/c mice against cutaneous leishmaniasis. Genetic susceptibility to L. major that results from induction of Th2 predominant immune responses after infection does not appear to reflect failure of skin DC to internalize or respond to parasites, or the inability of BALB/c T cells to mount a Th1 response to DC-associated Leishmania antigens. C1 NCI, Dermatol Branch, NCL, NIH, Bethesda, MD 20892 USA. NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. RP Udey, MC (reprint author), NCI, Dermatol Branch, NCL, NIH, Bldg 10,Rm 12N238, Bethesda, MD 20892 USA. OI Cushing, Melissa/0000-0001-8042-1494 NR 45 TC 87 Z9 90 U1 0 U2 1 PU WILEY-V C H VERLAG GMBH PI BERLIN PA PO BOX 10 11 61, D-69451 BERLIN, GERMANY SN 0014-2980 J9 EUR J IMMUNOL JI Eur. J. Immunol. PD DEC PY 2000 VL 30 IS 12 BP 3498 EP 3506 DI 10.1002/1521-4141(2000012)30:12<3498::AID-IMMU3498>3.0.CO;2-6 PG 9 WC Immunology SC Immunology GA 384YU UT WOS:000165975500019 PM 11093169 ER PT J AU Sehgal, D Schiaffella, E Anderson, AO Mage, RG AF Sehgal, D Schiaffella, E Anderson, AO Mage, RG TI Generation of heterogeneous rabbit anti-DNP antibodies by gene conversion and hypermutation of rearranged V-L and V-H genes during clonal expansion of B cells in splenic germinal centers SO EUROPEAN JOURNAL OF IMMUNOLOGY LA English DT Article DE rabbit; gene conversion; germinal center; immunoglobulin light chain; spleen ID SOMATIC DIVERSIFICATION; REGION SEQUENCES; COMBINING SITE; RECEPTOR; APPENDIX; MECHANISMS; EXPRESSION; REPERTOIRE; ALLOTYPES; DIVERSITY AB The mechanisms described here account for development of the heterogeneous high-affinity anti-DNP antibodies that rabbits can produce. Rearranged immunoglobulin light and heavy chain genes from single DNP-specific splenic germinal center B cells were amplified by PCR. We found that in clonal lineages, rearranged V-K and V-H are further diversified by gene conversion and somatic hypermutation. The positive and negative selection of amino acids in complementarity-determining regions observed allows emergence of a variety of different combining site structures. A by-product of the germinal center reaction may be cells with sequences altered by gene conversion that no longer react with the immunizing antigen but are a source of new repertoire. The splenic germinal center would thus play an additional role in adults similar to that of the appendix and other gut-associated lymphoid tissues of young rabbits. C1 NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. USA, Med Res Inst Infect Dis, Ft Detrick, MD 21702 USA. RP Mage, RG (reprint author), NIAID, Immunol Lab, NIH, Bldg 10,Rm 11N311,10 Ctr Dr,MSC 1892, Bethesda, MD 20892 USA. NR 40 TC 15 Z9 15 U1 0 U2 1 PU WILEY-V C H VERLAG GMBH PI BERLIN PA PO BOX 10 11 61, D-69451 BERLIN, GERMANY SN 0014-2980 J9 EUR J IMMUNOL JI Eur. J. Immunol. PD DEC PY 2000 VL 30 IS 12 BP 3634 EP 3644 DI 10.1002/1521-4141(200012)30:12<3634::AID-IMMU3634>3.0.CO;2-7 PG 11 WC Immunology SC Immunology GA 384YU UT WOS:000165975500035 PM 11169406 ER PT J AU Khaing, ZZ Weickert, CS Weinberger, DR Lipska, BK AF Khaing, ZZ Weickert, CS Weinberger, DR Lipska, BK TI Differential DNA damage in response to the neonatal and adult excitotoxic hippocampal lesion in rats SO EUROPEAN JOURNAL OF NEUROSCIENCE LA English DT Article DE animal model; cell death; ibotenic acid; schizophrenia ID METHYL-D-ASPARTATE; DEVELOPING CEREBRAL-CORTEX; CELL-DEATH; VENTRAL HIPPOCAMPUS; PREFRONTAL CORTEX; IMMATURE BRAIN; NEURONAL DEATH; APOPTOSIS; FRAGMENTATION; APOMORPHINE AB We examined the developmental profile of excitotoxin-induced nuclear DNA fragmentation using the transferase dUTP nick-end labelling (TUNEL) technique, as a marker of DNA damage and cell death in rats with neonatal and adult excitotoxic lesions of the ventral hippocampus. We hypothesized that infusion of neurotoxin may result in a differential pattern of cell death in neonatally and adult lesioned rats, both in the infusion site and in remote brain regions presumably involved in mediating behavioural changes observed in these animals. Brains of rats lesioned at 7 days of age and in adulthood were collected at several survival times 1-21 days after the lesion. In the lesioned neonates 1-3 days postlesion, marked increases in TUNEL-positive cells occurred in the ventral hippocampus, the site of neurotoxin infusion, and in a wide surrounding area. Adult lesioned brains showed more positive cells than controls only at the infusion site. In the lesioned neonates, TUNEL-labelled cells were also present in the striatum and nucleus accumbens I day postlesion but not at later survival times. Our findings indicate that cell death in remote regions is more prominent in immature than adult brains, that it may lead to distinct alterations in development of these brain:regions, and thus may be responsible for functional differences between neonatally and adult lesioned rats. C1 NIMH, IRP, Clin Brain Disorders Branch, Bethesda, MD 20892 USA. RP Lipska, BK (reprint author), NIMH, IRP, Clin Brain Disorders Branch, Bldg 10,Room 4N306, Bethesda, MD 20892 USA. RI Shannon Weickert, Cynthia/G-3171-2011; Lipska, Barbara/E-4569-2017 NR 47 TC 19 Z9 19 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0953-816X J9 EUR J NEUROSCI JI Eur. J. Neurosci. PD DEC PY 2000 VL 12 IS 12 BP 4424 EP 4433 DI 10.1046/j.0953-816X.2000.01320.x PG 10 WC Neurosciences SC Neurosciences & Neurology GA 387FV UT WOS:000166113000026 PM 11122353 ER PT J AU Pogun, S Demirgoren, S Taskiran, D Kanit, L Yilmaz, O Koylu, EO Balkan, B London, ED AF Pogun, S Demirgoren, S Taskiran, D Kanit, L Yilmaz, O Koylu, EO Balkan, B London, ED TI Nicotine modulates nitric oxide in rat brain SO EUROPEAN NEUROPSYCHOPHARMACOLOGY LA English DT Article DE nicotine; nitric oxide; glutamate; hippocampus; striatum; sex differences ID CANINE CEREBRAL-ARTERIES; L-ARGININE; SYNAPTIC TRANSMISSION; HIPPOCAMPAL SLICES; INDUCED RELAXATION; BIOLOGICAL-FLUIDS; SEX-DIFFERENCES; NERVE FUNCTION; H-3 DOPAMINE; CELL LOSS AB Nicotine exerts its central actions by regulating cationic fluxes through nicotinic acetylcholine receptors (nAChRs). By this effect, the drug likely also modifies events occurring beyond the nAChR, including the regulation of nitric oxide (NO) synthesis. The present study was undertaken to assess the effects of acute and chronic nicotine administration (0.4 mg/kg, s.c.) on levels of NO2- + NO3- stable metabolites of NO, in brain regions of male and female rats. Nicotine increased levels of the metabolites, and therefore presumably of NO, with sex differences in the degree of stimulation, the brain regions affected, and the variance between the effects of acute and chronic administration. Prior inhibition of NO synthase eliminated the effect of nicotine in all regions studied. While nicotine appeared to increase NO indirectly via glutamate receptors in the cortex and hippocampus, this was not true of the corpus striatum, where blocking NMDA-type glutamate receptors with MK-801 had no effect. The findings support the view that NO is likely involved in some of the central effects of nicotine. (C) 2000 Elsevier Science BN. All rights reserved. C1 Ege Univ, Ctr Brain Res, Sch Med, Dept Physiol, TR-35100 Izmir, Turkey. TUBITAK, Basic Neurosci Res Unit, TR-35100 Izmir, Turkey. NIDA, Brain Imaging Ctr, Intramural Res Program, NIH, Baltimore, MD 21224 USA. RP Pogun, S (reprint author), Ege Univ, Ctr Brain Res, Sch Med, Dept Physiol, TR-35100 Izmir, Turkey. RI Koylu, Ersin/A-4539-2011; Pogun, Sakire/A-5816-2010; OI Taskiran, Dilek/0000-0002-4505-0939 NR 60 TC 33 Z9 33 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0924-977X J9 EUR NEUROPSYCHOPHARM JI Eur. Neuropsychopharmacol. PD DEC PY 2000 VL 10 IS 6 BP 463 EP 472 DI 10.1016/S0924-977X(00)00116-4 PG 10 WC Clinical Neurology; Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA 388WM UT WOS:000166206100007 PM 11115736 ER PT J AU Mossner, R Albert, D Persico, AM Hennig, T Bengel, D Holtmann, B Schmitt, A Keller, F Simantov, R Murphy, D Seif, I Deckert, J Lesch, KP AF Mossner, R Albert, D Persico, AM Hennig, T Bengel, D Holtmann, B Schmitt, A Keller, F Simantov, R Murphy, D Seif, I Deckert, J Lesch, KP TI Differential regulation of adenosine A(1) and A(2A) receptors in serotonin transporter and monoamine oxidase A-deficient mice SO EUROPEAN NEUROPSYCHOPHARMACOLOGY LA English DT Article DE serotonin transporter; adenosine A(1) receptor; adenosine A(2A) receptor; dorsal raphe nucleus; accumbens nucleus; monoamine oxidase A; knockout mouse; autoradiography ID BRAIN-SEROTONIN; PANIC DISORDER; LACKING; LOCALIZATION; HIPPOCAMPUS; ACTIVATION; RELEASE; NUCLEUS; DISEASE; GENE AB The serotonin (5HT) transporter (5HTT) removes 5HT from the synaptic cleft and is thus critical to the control of serotonergic neurotransmission. Mice with a targeted inactivation of the 5HTT represent a novel and unique tool to study serotonergic system functioning. Because the release of 5HT is regulated by adenosine, we investigated 5HTT-deficient mice for possible adaptive changes of adenosine A(1) and A(2A) receptors. A(1) and A(2A) receptors were studied by means of quantitative autoradiography using the radioligands [H-3]8-cyclopentyl-1,3-dipropylxanthine and [H-3]CGS 21680, respectively. A comparison of 5HTT knockout versus control mice revealed upregulation of A(1) receptors in the dorsal raphe nucleus (DRN, +21%), but not in any of the serotonergic projection areas, and downregulation of A(2A) receptors in basal ganglia. The adaptive changes of A(1) and A(2A) receptors in 5HTT-deficient mice are likely to represent a compensatory neuroprotective effect mediated by the adenosinergic modulatory system. For comparison, these receptors were also studied in monoamine oxidase A (MAOA) knockout mice and in 5HTT/MAOA double knockout mice. 5HTT/MAOA double knockout mice showed adaptive changes of adenosine A(1) and A(2A) receptors similar to 5HTT knockout mice, while investigation of MAOA-deficient mice revealed an upregulation of A(2A) receptors, which may relate to a role of both MAOA and adenosine A(2A) receptors in anxiety. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Univ Wurzburg, Dept Psychiat & Psychotherapy, D-97080 Wurzburg, Germany. Neurosci Lab, I-00155 Rome, Italy. Univ Wurzburg, Dept Neurol, D-97078 Wurzburg, Germany. Weizmann Inst Sci, Dept Mol Genet, IL-76100 Rehovot, Israel. NIMH, Clin Sci Lab, Bethesda, MD 20892 USA. CNRS, UMR 146, F-91405 Orsay, France. Univ Munster, Dept Psychiat, D-48129 Munster, Germany. RP Mossner, R (reprint author), Univ Wurzburg, Dept Psychiat & Psychotherapy, Fuchsleinstr 15, D-97080 Wurzburg, Germany. RI Lesch, Klaus-Peter/J-4906-2013 OI Lesch, Klaus-Peter/0000-0001-8348-153X NR 20 TC 12 Z9 13 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0924-977X J9 EUR NEUROPSYCHOPHARM JI Eur. Neuropsychopharmacol. PD DEC PY 2000 VL 10 IS 6 BP 489 EP 493 DI 10.1016/S0924-977X(00)00119-X PG 5 WC Clinical Neurology; Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA 388WM UT WOS:000166206100010 PM 11115739 ER PT J AU Jucker, M Bondolfi, L Calhoun, ME Long, JM Ingram, DK AF Jucker, M Bondolfi, L Calhoun, ME Long, JM Ingram, DK TI Structural brain aging in inbred mice: potential for genetic linkage SO EXPERIMENTAL GERONTOLOGY LA English DT Article; Proceedings Paper CT 5th International Symposium on the Neurobiology and Neuroendocrinology of Aging CY JUL 23-28, 2000 CL BREGENZ, AUSTRIA DE aging; brain; CNS; neuron; glia; inclusions; mouse; strain; hippocampus; linkage; genetics; stereology; neurodegeneration; neurogenesis ID NEURODEGENERATIVE DISEASES; NEURONAL INCLUSIONS; MURINE MODELS; DENTATE GYRUS; C57BL/6 MICE; MOUSE; AGE; NEUROGENESIS; PHENOTYPES; THALAMUS AB To identify genetic factors involved in brain aging, we have initiated studies assessing behavioral and structural changes with aging among inbred mouse strains. Cognitive performance of C57BL/6J mice is largely maintained with aging, and stereological analysis revealed no significant age-related change in neuron number, synaptic bouton number or glial number in the hippocampus. Moreover, no change in cortical neuron number and cholinergic basal forebrain neuron number has been found in this strain. 129Sv/J mice have more pronounced age-related cognitive deficits, although hippocampal and basal cholinergic forebrain neuron number also appear unchanged with aging. Differences in neurogenesis and neuron vulnerability in the adult CNS of C57BL/6, 129/Sv and other inbred strains have been reported, which in turn may have important consequences for brain aging. Age-related lesions, such as thalamic eosinophilic inclusions and hippocampal clusters of polyglucosan bodies also vary greatly among inbred strains although the functional significance of these lesions is not clear. The continued assessment of such age-related structural and behavioral changes among inbred mouse strains offers the potential to identify genes that control age-related changes in brain structure and function. (C) 2000 Elsevier Science Inc. All rights reserved. C1 Univ Basel, Inst Pathol, CH-4003 Basel, Switzerland. Mt Sinai Sch Med, Neurobiol Aging Labs, New York, NY USA. NIA, Behav Neurosci Sect, Neurosci Lab, Gerontol Res Ctr,NIH, Baltimore, MD USA. RP Jucker, M (reprint author), Univ Basel, Inst Pathol, Schonbeinstr 40, CH-4003 Basel, Switzerland. NR 34 TC 20 Z9 21 U1 0 U2 3 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0531-5565 J9 EXP GERONTOL JI Exp. Gerontol. PD DEC PY 2000 VL 35 IS 9-10 SI SI BP 1383 EP 1388 DI 10.1016/S0531-5565(00)00190-X PG 6 WC Geriatrics & Gerontology SC Geriatrics & Gerontology GA 384XQ UT WOS:000165972900022 PM 11113616 ER PT J AU Ikonomi, P Rivera, CE Riordan, M Washington, G Schechter, AN Noguchi, CT AF Ikonomi, P Rivera, CE Riordan, M Washington, G Schechter, AN Noguchi, CT TI Overexpression of GATA-2 inhibits erythroid and promotes megakaryocyte differentiation SO EXPERIMENTAL HEMATOLOGY LA English DT Article DE GATA-2; erythroid progenitors; K562; megakaryocyte; differentiation ID TRANSCRIPTION FACTOR; MEGAKARYOBLASTIC DIFFERENTIATION; HEMATOPOIETIC DEFECT; BINDING PROTEINS; GENE-EXPRESSION; MESSENGER-RNA; CELL-LINES; STEM-CELLS; LEUKEMIA; EOSINOPHILS AB Objective. GATA-1 and GATA-2 transcription factors are required for effective hematopoiesis. These regulatory proteins present overlapping yet distinct patterns of expression in hematopoietic cells. Absence of GATA-2 leads to defective hematopoiesis and an embryonic lethal phenotype. Disruption of GATA-1 results in a compensatory increase in GATA-2 in early erythroid cells and incomplete erythropoiesis with embryos dying at 11.5 days. We examine the specific role of GATA-2 later in hematopoiesis, during erythroid differentiation. Materials and Methods. Stable K562 cell lines expressing various levels of GATA-2 were generated using a GATA-2 expression plasmid, Overexpression of GATA-2 transcripts was determined by quantitative polymerase chain reaction (PCR). Cytospin smears, growth curve analysis, PCR, and flow cytometry were used to examine the effects of increased levels of GATA-2 in altering cell phenotype and activation of megakaryocytic markers. Human progenitor erythroid cells also were transfected with a GATA-2 expression vector. Growth curve analysis, benzidine staining, and high-performance liquid chromatographic analysis were used to study the effects of GATA-2 on erythroid maturation and proliferation. Results. K562/GATA-2 cell lines expressing high levels of GATA-2 mRNA showed a marked decrease in proliferation and a shift in phenotype toward the megakaryocyte lineage. Ploidy analyses showed that these cell lines developed a multinuclear phenotype, including tetraploids and octaploids. PCR analysis showed activation of megakaryocyte-specific genes including thrombopoietin receptor (c-mpl). Surface expression of platelet glycoprotein receptors Ib/IX (CD42b/CD42a) and IIb/IIIa (CD41/CD61) also was demonstrated by flow cytometry. In primary human adult erythroid cultures transfected with a GATA-2 expression vector, production of total hemoglobin and cell proliferation decreased in a dose-dependent manner. Conclusions. GATA-2 plays an important role in deciding cell lineage throughout hematopoiesis, and increased expression of GATA-2 determines megakaryocytic differentiation. Downregulation of GATA-2 is required for erythroid differentiation. (C) 2000 International Society for Experimental Hematology. Published by Elsevier Science Inc. C1 NIDDK, Biol Chem Lab, NIH, Bethesda, MD 20892 USA. NIH, Hematol Serv, Dept Clin Pathol, Ctr Clin, Bethesda, MD USA. US FDA, Div Cellular & Gene Therapies, Ctr Biol Evaluat & Res, Rockville, MD 20857 USA. RP Noguchi, CT (reprint author), NIDDK, Biol Chem Lab, NIH, Bldg 10,Room 9N-307,10 Ctr Dr,MSC 1822, Bethesda, MD 20892 USA. OI Schechter, Alan N/0000-0002-5235-9408 NR 34 TC 64 Z9 71 U1 2 U2 3 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0301-472X J9 EXP HEMATOL JI Exp. Hematol. PD DEC PY 2000 VL 28 IS 12 BP 1423 EP 1431 DI 10.1016/S0301-472X(00)00553-1 PG 9 WC Hematology; Medicine, Research & Experimental SC Hematology; Research & Experimental Medicine GA 385XB UT WOS:000166029500016 PM 11146164 ER PT J AU Chian, R Young, S Danilkovitch-Miagkova, A Ronnstrand, L Leonard, E Ferrao, P Ashman, L Linnekin, D AF Chian, R Young, S Danilkovitch-Miagkova, A Ronnstrand, L Leonard, E Ferrao, P Ashman, L Linnekin, D TI P13 kinase mediates transformation of hematopoietic cells by the V816 c-Kit mutant SO EXPERIMENTAL HEMATOLOGY LA English DT Meeting Abstract C1 NCI, Frederick Canc Res & Dev Ctr, BRL, Frederick, MD USA. NCI, Frederick Canc Res & Dev Ctr, LI, DBS, Frederick, MD USA. Hanson Ctr Canc Res, Adelaide, SA, Australia. Univ Adelaide, Adelaide, SA, Australia. Biomedicum, Ludwig Inst Canc Res, Uppsala, Sweden. RI Ronnstrand, Lars/A-2429-2011; ASHMAN, LEONIE/G-7631-2013 OI ASHMAN, LEONIE/0000-0003-3559-3611 NR 0 TC 5 Z9 5 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0301-472X J9 EXP HEMATOL JI Exp. Hematol. PD DEC PY 2000 VL 28 IS 12 MA 901 BP 1491 EP 1491 DI 10.1016/S0301-472X(00)00562-2 PG 1 WC Hematology; Medicine, Research & Experimental SC Hematology; Research & Experimental Medicine GA 385XB UT WOS:000166029500026 ER PT J AU Sredni, B Longo, DL Catane, R Shani, A Albeck, M Kalechman, Y AF Sredni, B Longo, DL Catane, R Shani, A Albeck, M Kalechman, Y TI Synergistic antitumoral effect of the immunomodulator AS101+paclitaxel (Taxol): Association with ras-dependent signal transduction pathways SO EXPERIMENTAL HEMATOLOGY LA English DT Meeting Abstract C1 Bar Han Univ, Fac Life Sci, CAIR Inst, Ramat Gan, Israel. NIA, NIH, Baltimore, MD 21224 USA. Shaare Zedek Med Ctr, Jerusalem, Israel. Kaplan Hosp, IL-76100 Rehovot, Israel. NR 0 TC 1 Z9 1 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0301-472X J9 EXP HEMATOL JI Exp. Hematol. PD DEC PY 2000 VL 28 IS 12 MA 916 BP 1496 EP 1496 DI 10.1016/S0301-472X(00)00630-5 PG 1 WC Hematology; Medicine, Research & Experimental SC Hematology; Research & Experimental Medicine GA 385XB UT WOS:000166029500040 ER PT J AU Lubet, RA Zhang, ZQ Wiseman, RW You, M AF Lubet, RA Zhang, ZQ Wiseman, RW You, M TI Use of p53 transgenic mice in the development of cancer models for multiple purposes SO EXPERIMENTAL LUNG RESEARCH LA English DT Article; Proceedings Paper CT 3rd International Mouse Lung Tumorigenesis Symposium CY JUN 16-17, 2000 CL MED COLL OHIO, TOLEDO, OHIO SP Ohio State Univ, NCI, Natl Inst Environm Hlth Sci, Proctor & Gamble, Novartis HO MED COLL OHIO DE cancer; carcinogenesis; chemoprevention; chemotherapy; colon; lung; p53 ID METHYL-N-NITROSOUREA; INDUCED LUNG-TUMORS; KI-RAS; MUTATIONS; CARCINOGENESIS; CARCINOMAS; GENE; N-BUTYL-N-(4-HYDROXYBUTYL)NITROSAMINE; PROGRESSION; MOUSE AB The tumor suppressor gene p53 is perhaps the most commonly mutated gene in human cancer, being mutated in a high percentage of colon, breast, skin, bladder, and many cancers of the aerodigestive tract. Individuals with Li-Fraumeni syndrome, who routinely have a germline mutation in the p53 tumor suppressor gene, are at high risk for lung cancer, confirming its intimate role in lung tumorigenesis in humans. In contrast, the majority of chemically induced or spontaneous cancers in rodents do not contain mutations in p53. Therefore, we examined a transgenic mouse that contains a dominant negative mutation (Arg135Val) in the p53 gene placed under the control of its own endogenous promoter. The resulting mice have 3 copies of the mutated transgene as well as 2 normal p53 alleles. In the chemical carcinogenesis studies, we employed mice containing the mutated p53 gene to examine for carcinogen susceptibility. We found that mice with the p53 mutation, on an A/J F1 background, were more susceptible to a number of potential lung carcinogens, including N-methyl-N-nitrosourea (MNU) and the known tobacco carcinogens 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and benzo(a)pyrene (BP). Mice with a mutant p53 developed larger tumors and roughly 3 times as many tumors, emphasizing the potential effects of a p53 mutation both on tumor initiation and progression. In addition, we examined 2 nonlung carcinogens, 1,2-dimethylhydrazine (DMH), a colon carcinogen, and N-butyl-N-(4-hydroxybutyl)-nitrosamine (OHBBN), a bladder carcinogen. Interestingly a germline p53 mutation increased the incidence of DMH-induced colon, lung, hepatic, and uterine tumors, while having limited effects on OHBBN-induced bladder tumors. Because of its heightened susceptibility we are examining the use of this model in smoke-induced tumorigenesis in A/J mice as well. Employing the lung adenomas induced by NNK, we found that mice with or without a p53 mutation were equally susceptible to the chemopreventive effects of dexamethasone plus myo-inisitol and green tea. The tumors, which arise in a highly reproducible manner in p53 transgenic mice following carcinogen treatment, have mutations in both p53 and the K-ras oncogene. Thus, this model appears useful for examining for potential chemotherapeutic agents, p53 mutated or wild-type mice were equally susceptible to the therapeutic effects of Taxol or Adriamycin. Interestingly, piroxicam was similarly effective in inhibiting colon tumor formation by DMH in mice with or without a mutation in the p53 tumor suppressor gene. In contrast, lung and uterine tumors developing in these mice were not susceptible to the chemopreventive effects of piroxicam. In summary, mice with mutations in the p53 tumor suppressor gene appear to be particularly applicable for basic mechanistic studies, for screening for potential carcinogens, and for screening for chemopreventive or chemotherapeutic agents. C1 NCI, CADRG, Chemoprevent Branch, Bethesda, MD 20852 USA. Ohio State Univ, James Canc Ctr, Columbus, OH 43210 USA. RP Lubet, RA (reprint author), NCI, CADRG, Chemoprevent Branch, Execut Plaza N,Room 201,6130 Execut Blvd, Bethesda, MD 20852 USA. NR 27 TC 24 Z9 26 U1 0 U2 7 PU HEMISPHERE PUBL CORP PI BRISTOL PA 1900 FROST ROAD, SUITE 101, BRISTOL, PA 19007-1598 USA SN 0190-2148 J9 EXP LUNG RES JI Exp. Lung Res. PD DEC PY 2000 VL 26 IS 8 BP 581 EP 593 PG 13 WC Respiratory System SC Respiratory System GA 388WH UT WOS:000166205700003 PM 11195457 ER PT J AU Linnoila, RI Sahu, A Miki, M Ball, DW DeMayo, FJ AF Linnoila, RI Sahu, A Miki, M Ball, DW DeMayo, FJ TI Morphometric analysis of CC10-hASH1 transgenic mouse lung: A model for bronchiolization of alveoli and neuroendocrine carcinoma SO EXPERIMENTAL LUNG RESEARCH LA English DT Article; Proceedings Paper CT 3rd International Mouse Lung Tumorigenesis Symposium CY JUN 16-17, 2000 CL MED COLL OHIO, TOLEDO, OHIO SP Ohio State Univ, NCI, Natl Inst Environm Hlth Sci, Proctor & Gamble, Novartis HO MED COLL OHIO DE bronchioloalveolar metaplasia; human achaete-scute homolog-1 (hASH1); lung; carcinogenesis; morphometric analysis; neuroendocrine tumors; Simian virus 40 large T antigen (TAg); transgenic mice ID SV40 T-ANTIGEN; DIFFERENTIATION; EXPRESSION; CANCER; CELLS; MORPHOGENESIS; OZONE; MICE AB Constitutive expression of human achaete-scute homolog-1 (hASH1) alone or in combination with Simian virus 40 (SV40) large T antigen (TAg) under the Clara cell 10-kDa secretory protein (CC10) promoter results in bronchiolization of alveoli and enhanced tumorigenesis, respectively. A novel morphometric system composed of series of fixed distance concentric rings originating at the bronchioloalveolar junction was used to determine spatial growth patterns. hASH1 mice exhibited progressive airway epithelial hyperplasia near this junction, and minimal changes further away in the alveolar compartment. TAg animals shared this increase, but exhibited variable distance-dependent growth. By 2 months TAg/hASH1 animals showed highly increased growth at all points measured. Remarkably, TAg/hASH1 animals expressed both CC10 and extensive neuroendocrine differentiation in airways and tumors. The results suggest that these transgenic mice provide a useful model with many similarities to human lung carcinogenesis, which originates in airway epithelium, and often reveals neuroendocrine differentiation. C1 NCI, Cell & Canc Biol Dept, Med Branch, Div Clin Sci,NIH, Rockville, MD 20850 USA. Johns Hopkins Univ, Sch Med, Ctr Oncol, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Dept Med, Baltimore, MD 21205 USA. Baylor Univ, Sch Med, Dept Mol & Cell Biol, Houston, TX 77030 USA. RP Linnoila, RI (reprint author), NCI, Cell & Canc Biol Dept, Med Branch, Div Clin Sci,NIH, 9610 Med Ctr Dr,Suite 300, Rockville, MD 20850 USA. NR 22 TC 18 Z9 19 U1 0 U2 1 PU HEMISPHERE PUBL CORP PI BRISTOL PA 1900 FROST ROAD, SUITE 101, BRISTOL, PA 19007-1598 USA SN 0190-2148 J9 EXP LUNG RES JI Exp. Lung Res. PD DEC PY 2000 VL 26 IS 8 BP 595 EP 615 PG 21 WC Respiratory System SC Respiratory System GA 388WH UT WOS:000166205700004 PM 11195458 ER PT J AU Zhang, ZQ Lin, L Liu, GJ Wang, M Hill, J Wang, Y You, M Devereux, TR AF Zhang, ZQ Lin, L Liu, GJ Wang, M Hill, J Wang, Y You, M Devereux, TR TI Fine mapping and characterization of candidate lung tumor resistance genes for the Par2 locus on mouse chromosome 18 SO EXPERIMENTAL LUNG RESEARCH LA English DT Article; Proceedings Paper CT 3rd International Mouse Lung Tumorigenesis Symposium CY JUN 16-17, 2000 CL MED COLL OHIO, TOLEDO, OHIO SP Ohio State Univ, NCI, Natl Inst Environm Hlth Sci, Proctor & Gamble, Novartis HO MED COLL OHIO DE chromosome 18; genetics; lung tumor susceptibility; mouse; pulmonary adenoma resistance gene ID IN-VIVO ALTERATIONS; DCC-GENE; PULMONARY ADENOMA; SUSCEPTIBILITY LOCUS; DPC4 GENE; CANCER; MICE; EXPRESSION; CARCINOMA; RECEPTOR AB In number of recent studies, a lung tumor resistance locus designated either Par2 or Pas7 was mapped to distal chromosome 18 in crosses between susceptible A/J and more resistant BALB/c mice. This locus is important in that it accounts for as much as 60% of the difference in lung tumor susceptibility between the A/J and BALB/c mice, both of which contain the susceptible allele of Kras2, a marker and strong candidate for the major lung tumor susceptibility gene on mouse chromosome 6. We have now fine-mapped the Par2 locus by using congenic mice that were constructed by placing part of chromosome 18 from the susceptible A/J onto the genetic background of lung tumor-resistant BALB/c mice. After 7 generations of backcrossing, N7 mice that carried 28 cM of the A/J quantitative trait locus (QTL) region were crossed to the BALB/c to generate the N8 generation. Congenic strains (N8) that contain various QTL regions were generated. N9 mice, generated from N8 males x 3 BALB/c females, were genotyped in the region of the Par2 locus and treated with an initiating dose of urethane and allowed to form lung tumors over 6 months. The mice were killed and the lung tumors counted. With this cross the Par2 locus was narrowed to a 6-cM region. Potential candidate genes in this region include Smad4, Smad2 and Dcc. Previously, we excluded Smad4 and Smad2 as candidates for Par2 based on the lack of functional polymorphism(s) and differential expression in lungs from A/J and BALB/c mice. In this study, no polymorphism of the coding sequence of Dcc was observed between A/J and BALB/c mice. Further, fine mapping and positional cloning are required for the identification of the Par2 gene. C1 NIEHS, Mol Carcinogenesis Lab, Res Triangle Pk, NC 27709 USA. Ohio State Univ, James Canc Ctr, Div Human Canc Genet, Columbus, OH 43210 USA. Med Coll Ohio, Dept Pathol, Toledo, OH 43699 USA. RP Devereux, TR (reprint author), NIEHS, Mol Carcinogenesis Lab, Mail Drop D4-04,POB 12233, Res Triangle Pk, NC 27709 USA. FU NCI NIH HHS [CA78797, CA58554] NR 31 TC 9 Z9 9 U1 0 U2 1 PU HEMISPHERE PUBL CORP PI BRISTOL PA 1900 FROST ROAD, SUITE 101, BRISTOL, PA 19007-1598 USA SN 0190-2148 J9 EXP LUNG RES JI Exp. Lung Res. PD DEC PY 2000 VL 26 IS 8 BP 627 EP 639 PG 13 WC Respiratory System SC Respiratory System GA 388WH UT WOS:000166205700006 PM 11195460 ER PT J AU Liu, JF Gu, P Bergman, G Kelloff, GJ Boone, CW You, M Wang, Y AF Liu, JF Gu, P Bergman, G Kelloff, GJ Boone, CW You, M Wang, Y TI Detection of genetic alterations tn mouse lung adenocarcinomas by two-dimensional gel electrophoresis SO EXPERIMENTAL LUNG RESEARCH LA English DT Article; Proceedings Paper CT 3rd International Mouse Lung Tumorigenesis Symposium CY JUN 16-17, 2000 CL MED COLL OHIO, TOLEDO, OHIO SP Ohio State Univ, NCI, Natl Inst Environm Hlth Sci, Proctor & Gamble, Novartis HO MED COLL OHIO DE 2-dimensional gel electrophoresis; genetic alterations; lung tumors; mouse ID TUMOR-SUPPRESSOR GENE; DNA METHYLATION; CANCER; CHROMOSOME-4 AB DNA from 14 mouse lung adenocarcinomas and 7 normal lungs were examined by 2- dimensional gel electrophoresis (2-DGE) for genetic alterations. 2-DGE profiles from tumor samples were compared with those profiles from normal lung tissues through a computer-assisted color overlay system. Compared to the profiles in normal lung DNA, 6 spots were amplified and 16 spots were partially reduced in their intensity in tumors. Two spots were detectable only in tumor tissues. These altered spots indicate genetic changes in mouse lung tumor development. The identification of these genetic alterations is probably important in understanding mouse lung carcinogenesis. C1 Ohio State Univ, Sch Publ Hlth, James Canc Ctr, Columbus, OH 43210 USA. Med Coll Ohio, Dept Pathol, Toledo, OH 43699 USA. NCI, Chemoprevent Branch, NIH, Bethesda, MD 20892 USA. RP Wang, Y (reprint author), Ohio State Univ, Sch Publ Hlth, James Canc Ctr, 1148 CHRI,300 W 10th Ave, Columbus, OH 43210 USA. FU NCI NIH HHS [CA58554] NR 19 TC 1 Z9 1 U1 0 U2 0 PU HEMISPHERE PUBL CORP PI BRISTOL PA 1900 FROST ROAD, SUITE 101, BRISTOL, PA 19007-1598 USA SN 0190-2148 J9 EXP LUNG RES JI Exp. Lung Res. PD DEC PY 2000 VL 26 IS 8 BP 651 EP 658 PG 8 WC Respiratory System SC Respiratory System GA 388WH UT WOS:000166205700008 PM 11195462 ER PT J AU Ramakrishna, G Sithanandam, G Cheng, RYS Fornwald, LW Smith, GT Diwan, BA Anderson, LM AF Ramakrishna, G Sithanandam, G Cheng, RYS Fornwald, LW Smith, GT Diwan, BA Anderson, LM TI K-ras p21 expression and activity in lung and lung tumors SO EXPERIMENTAL LUNG RESEARCH LA English DT Article; Proceedings Paper CT 3rd International Mouse Lung Tumorigenesis Symposium CY JUN 16-17, 2000 CL MED COLL OHIO, TOLEDO, OHIO SP Ohio State Univ, NCI, Natl Inst Environm Hlth Sci, Proctor & Gamble, Novartis HO MED COLL OHIO DE cell division; Erk; growth arrest; K-ras; raf; tumor size ID ALVEOLAR EPITHELIAL-CELLS; GENE-EXPRESSION; GROWTH-FACTOR; ACTIVATION; MUTATIONS; ONCOGENES; PROTEINS; RECEPTOR; ALLELE; MICE AB Although K-ras is mutated in many human and mouse lung adenocarcinomas, the function of K-ras p21 in lung is not known. We sought evidence for the prevalent hypothesis that K-ras p21 activates raf, which in turn passes the signal through the extracellular signal regulated Kinases (Erks) to stimulate cell division, and that this pathway is upregulated when K-ras is mutated. Results from both mouse lung tumors and immortalized cultured E10 and C10 lung type II cells failed to substantiate this hypothesis. Lung tumors did not have more total K-ras p21 or K-ras p21 GTP than normal lung tissue, nor were high levels of these proteins found in tumors with mutant K-ras. Activated K-ras p21-GTP levels did not correlate with proliferating cell nuclear antigen. Special features of tumors with mutant K-ras included small size of carcinomas compared with carcinomas lacking this mutation, and correlation of proliferating cell nuclear antigen with raf-1. In nontransformed type II cells in culture, both total and activated K-ras p21 increased markedly at confluence but not after serum stimulation, whereas both Erk1/2 and the protein kinase AKt were rapidly activated by the serum treatment. Reverse transcriptase-polymerase chain reaction (RT-PCR) assays of K-ras mRNA indicated an increase in confluent and especially in postconfluent cells. Together the findings indicate that normal K-ras p21 activity is associated with growth arrest of lung type II cells, and that the exact contribution of mutated K-ras p21 to tumor development remains to be discovered. C1 NCI, Comparat Carcinogenesis Lab, Frederick, MD 21701 USA. SAIC Frederick Inc, Frederick Canc Res & Dev Ctr, Frederick, MD 21701 USA. RP Anderson, LM (reprint author), Bldg 538,Room 205 B,Ft Detrick, Frederick, MD 21702 USA. NR 39 TC 18 Z9 19 U1 1 U2 3 PU HEMISPHERE PUBL CORP PI BRISTOL PA 1900 FROST ROAD, SUITE 101, BRISTOL, PA 19007-1598 USA SN 0190-2148 J9 EXP LUNG RES JI Exp. Lung Res. PD DEC PY 2000 VL 26 IS 8 BP 659 EP 671 DI 10.1080/01902140150216747 PG 13 WC Respiratory System SC Respiratory System GA 388WH UT WOS:000166205700009 PM 11195463 ER PT J AU Kang, Y Prentice, MA Mariano, JM Davarya, S Linnoila, RI Moody, TW Wakefield, LM Jakowlew, SB AF Kang, Y Prentice, MA Mariano, JM Davarya, S Linnoila, RI Moody, TW Wakefield, LM Jakowlew, SB TI Transforming growth factor-beta 1 and its receptors in human lung cancer and mouse lung carcinogenesis SO EXPERIMENTAL LUNG RESEARCH LA English DT Article; Proceedings Paper CT 3rd International Mouse Lung Tumorigenesis Symposium CY JUN 16-17, 2000 CL MED COLL OHIO, TOLEDO, OHIO SP Ohio State Univ, NCI, Natl Inst Environm Hlth Sci, Proctor & Gamble, Novartis HO MED COLL OHIO DE carcinogenesis; human; lung cancer; mouse; receptor; TGF-beta ID TGF-BETA-RECEPTOR; II RECEPTOR; CELL-LINES; PULMONARY ADENOCARCINOMA; HUMAN BREAST; DOWN-REGULATION; MCF-7 CELLS; EXPRESSION; CARCINOMA; PROTEIN AB The transforming growth factor-betas (TGF-betas) are multifunctional proteins that inhibit the proliferation of many epithelial cells through a set of cell protein receptors that includes the TGF-beta type I (RI) and type II (RII) receptors. Loss of growth inhibition by TGF-beta is thought to contribute to the development of many types of tumors. In the present study, we have examined expression of the proteins and mRNAs for TGF-beta1, TGF-beta RI, and TGF-beta RII in normal human lung, well-characterized non-small cell lung cancer (NSCLC) cell lines, and primary NSCLC specimens. Immunohistochemical staining for TGF-beta1, TGF-beta RI, and TGF-beta RII using specific antibodies in normal human lung showed expression of the 3 proteins in the epithelium of bronchi and bronchioles as well as in alveoli. Differential expression of TGF-beta RI and TGF-beta RII proteins was detected in 5 NSCLC cell lines using Western blot analysis, with reduced levels in 3 cell lines. A panel of 45 formalin-fixed and paraffin-embedded NSCLC specimens showed positive immunostaining for TGF-beta1, TGF-beta RI, and TGF-beta RII, with reduced TGF-beta RII in poorly differentiated adenocarcinomas and squamous cell carcinomas and some moderately differentiated adenocarcinomas. In situ hybridization studies conducted with specific riboprobes for TGF-beta1, TGF-beta RI, and TGF-beta RII showed corresponding localization of expression of the mRNAs in the specimens that showed positive immunostaining for the proteins. To investigate the roles of TGF-beta1, TGF-beta RI, and TGF-beta RII in chemically induced mouse lung tumorigenesis, we examined the expression of their proteins and mRNAs in 2 mouse model systems. Whereas expression of the proteins and mRNAs for TGF-beta1 and TGF-beta RI was comparable in lung adenomas and bronchioles of A/J mice treated with benzo(alpha )pyrene, decreased immunostaining and hybridization for TGF-beta RII protein and mRNA was detected in 50% of lung adenomas in these mice. Interestingly, expression of TGF-beta1 and the TGF-beta receptor proteins was similar to that of bronchioles in C57B1/6 mice and their littermates heterozygous for deletion of the TGF-beta1 gene treated with diethylnitrosamine. These data show that reduced levels of expression of TGF-beta RII occur in some, but not all, human and mouse lung tumors. This suggests that different mechanisms of action, some of which may involve the TGF-beta signaling pathway, may contribute to the progression of lung tumorigenesis. C1 NCI, Med Branch, Dept Cell & Canc Biol, Rockville, MD 20850 USA. NCI, Lab Cell Regulat & Carcinogenesis, Bethesda, MD 20892 USA. RP Jakowlew, SB (reprint author), NCI, Med Branch, Dept Cell & Canc Biol, 9610 Med Ctr Dr,Suite 300, Rockville, MD 20850 USA. NR 48 TC 21 Z9 24 U1 0 U2 1 PU HEMISPHERE PUBL CORP PI BRISTOL PA 1900 FROST ROAD, SUITE 101, BRISTOL, PA 19007-1598 USA SN 0190-2148 J9 EXP LUNG RES JI Exp. Lung Res. PD DEC PY 2000 VL 26 IS 8 BP 685 EP 707 PG 23 WC Respiratory System SC Respiratory System GA 388WH UT WOS:000166205700011 PM 11195465 ER PT J AU Gunning, WT Kramer, PM Lubet, RA Steele, VE Pereira, MA AF Gunning, WT Kramer, PM Lubet, RA Steele, VE Pereira, MA TI Chemoprevention of vinyl carbamate-induced lung tumors in strain a mice SO EXPERIMENTAL LUNG RESEARCH LA English DT Article; Proceedings Paper CT 3rd International Mouse Lung Tumorigenesis Symposium CY JUN 16-17, 2000 CL MED COLL OHIO, TOLEDO, OHIO SP Ohio State Univ, NCI, Natl Inst Environm Hlth Sci, Proctor & Gamble, Novartis HO MED COLL OHIO DE carcinogenesis; chemoprevention; dexamethasone; DMSO; green tea; lung tumors; piroxicam ID FEMALE A/J MICE; CLARA CELL ADENOMA; GREEN TEA; ETHYL CARBAMATE; MOUSE LUNG; BLACK TEA; N-NITROSODIETHYLAMINE; TOBACCO-SMOKE; ALPHA-DIFLUOROMETHYLORNITHINE; PULMONARY CARCINOGENESIS AB The ability of potential chemopreventive agents to prevent vinyl carbamate-induced lung turners was determined in 2 different experiments. Female strain A mice adminstered intraperitoneally either a single injection of 60 mg/kg vinyl carbamate that induced 24.0 +/- 1.72 tumors/mouse at 24 weeks or 2 injections of 16 mg/kg vinyl carbamate each (32 mg/kg total dose) that induced 43.2 +/- 3.2 tumors/mouse at 20 weeks. Lung carcinomas were Sound as early as 16 weeks. Dexamethasone and piroxicam provided in the diet were found to significantly inhibit lung tumors induced by 60 mg/kg vinyl carbamate at 24 weeks whereas myo-inositol also provided in the diet, did not significantly inhibit tumor formation. In animals given 6 16-mg/kg doses of vinyl carbamate, tumor multiplicity was reduced roughly 25% by alpha -difluoromethylornithine and green tea and reduced 50% by dexamethasone and piroxicam. Combinations of these agents were also tested using a total dose of 32 mg/kg of vinyl carbamate. Although alpha -difluoromethylornithine and green tea did not result in a significant inhibition of lung tumor formation if used alone, the combination of alpha -difluoromethylornithine and green tea resulted in a significant reduction of tumor multiplicity. The combinations of alpha -difluoromethylornithine or green tea with either dexamethasone or piroxicam or the combination of dexamethasone and piroxicam did not decrease tumor multiplicity greater than achieved by dexamethasone and piroxicam alone. In summary, selected chemopreventive agents previously shown to inhibit lung tumors by other chemical carcinogens also inhibited vinyl carbamate-induced lung tumors. C1 Med Coll Ohio, Dept Pathol, Toledo, OH 43614 USA. NCI, Div Canc Prevent, Bethesda, MD 20892 USA. RP Gunning, WT (reprint author), Med Coll Ohio, Dept Pathol, Block Hlth Sci Bldg,3035 Arlington Ave, Toledo, OH 43614 USA. RI Gunning, William/E-4681-2010 FU NCI NIH HHS [N01-CN-75104, N01-CN-75102] NR 51 TC 33 Z9 33 U1 0 U2 2 PU HEMISPHERE PUBL CORP PI BRISTOL PA 1900 FROST ROAD, SUITE 101, BRISTOL, PA 19007-1598 USA SN 0190-2148 J9 EXP LUNG RES JI Exp. Lung Res. PD DEC PY 2000 VL 26 IS 8 BP 757 EP 772 PG 16 WC Respiratory System SC Respiratory System GA 388WH UT WOS:000166205700015 PM 11195469 ER PT J AU Lantry, LE Zhang, ZQ Crist, KA Wang, Y Hara, M Zeeck, A Lubet, RA You, M AF Lantry, LE Zhang, ZQ Crist, KA Wang, Y Hara, M Zeeck, A Lubet, RA You, M TI Chemopreventive efficacy of promising farnesyltransferase inhibitors SO EXPERIMENTAL LUNG RESEARCH LA English DT Article; Proceedings Paper CT 3rd International Mouse Lung Tumorigenesis Symposium CY JUN 16-17, 2000 CL MED COLL OHIO, TOLEDO, OHIO SP Ohio State Univ, NCI, Natl Inst Environm Hlth Sci, Proctor & Gamble, Novartis HO MED COLL OHIO DE chemoprevention; farnesyltransferase; lung tumors; ras ID PROTEIN TRANSFERASE BLOCKS; SELECTIVE-INHIBITION; PERILLYL ALCOHOL; RAS ONCOGENES; MOLECULAR ANALYSIS; HUMAN CANCER; TRANSFORMATION; FARNESYL; CELLS; DEHYDROEPIANDROSTERONE AB The studies presented were designed to test the efficacy of farnesyltransferase inhibitors (FTIs) as potential chemopreventive compounds in the mouse lung tumor model, and in tumor cell lines. The compounds included manumycin, gliotoxin, dihydroepiandrosterone (DHEA) perillyl alcohol (POH), and FTI-276. Each of these compounds had the potential, based on in vitro and limited in vivo evidence, to inhibit mouse lung tumorigenesis. In vitro studies were conducted with both K-ras-transformed NIH-3T3 cells and mouse lung tumor epithelial cell lines. We utilized 2 primary mouse lung tumor models that reliably produce lung tumors with an oncogenic K-ras mutation when induced by 4-(methylnitrosoamino)-1-(3-pyridyl)-1-butanone (NNK). Manumycin, gliotoxin, DHEA, and POH were administered 3 times per week peritoneally (IP), starting 1 week prior to carcinogen treatment, and throughout the test period (4.5 months). FTI-276 was delivered daily for 4 months by a time-release pellet method. Both the manumycin and gliotoxin treatment groups demonstrated 100% incidence and an increase in tumor multiplicity over control, of 66% and 58% increase respectively (P < .05). Although DHEA showed no significant chemopreventive effect, POH treatment demonstrated a 22% reduction in tumor incidence (P < .05) and a 58% reduction in tumor multiplicity (P < .05). Finally, FTI-276 reduced both the tumor multiplicity by 41.7% (P < .005), and the total tumor volume/burden per mouse by 79.4% (P < .0001). The apoptotic index in FTI-276-treated tumors showed an increase of 77% over control tumors (P < .05). In vitro, all compounds demonstrated growth inhibition at a dose-response manner; however, manumycin, gliotoxin, and DHEA demonstrated an initial increase in growth rate at lower doses. In summary, we have shown that POH and FTI-276 are chemopreventive in a primary mouse lung tumor model. In contrast, DHEA was not significantly chemopreventive at the dosage utilized and treatment of an immunocompetent host with manumycin or gliotoxin demonstrated a significant increase in tumorigenicity over carcinogen control. C1 Ohio State Univ, James Canc Ctr, Div Human Canc Genet, Columbus, OH 43210 USA. Med Coll Ohio, Dept Pathol & Surg, Toledo, OH 43699 USA. Kyowa Hakko Kogyo Co Ltd, Tokyo Res Labs, Tokyo 194, Japan. Univ Gottingen, Inst Organ Chem, D-3400 Gottingen, Germany. NCI, Chemoprevent Branch, Rockville, MD USA. RP You, M (reprint author), Ohio State Univ, James Canc Ctr, Div Human Canc Genet, 646 Med Res Facil,420 W 12th Ave, Columbus, OH 43210 USA. FU NCI NIH HHS [CN55184, CA58554] NR 39 TC 19 Z9 19 U1 0 U2 2 PU HEMISPHERE PUBL CORP PI BRISTOL PA 1900 FROST ROAD, SUITE 101, BRISTOL, PA 19007-1598 USA SN 0190-2148 J9 EXP LUNG RES JI Exp. Lung Res. PD DEC PY 2000 VL 26 IS 8 BP 773 EP 790 PG 18 WC Respiratory System SC Respiratory System GA 388WH UT WOS:000166205700016 PM 11195470 ER PT J AU Chang, CF Niu, KC Hoffer, BJ Wang, Y Borlongan, CV AF Chang, CF Niu, KC Hoffer, BJ Wang, Y Borlongan, CV TI Hyperbaric oxygen therapy for treatment of postischemic stroke in adult rats SO EXPERIMENTAL NEUROLOGY LA English DT Article DE cerebral ischemia; reperfusion; hyperoxygenation; hyperbaric pressure; plasma pH; arterial pressure; motor asymmetry ID MIDDLE CEREBRAL-ARTERY; HUNTINGTONS-DISEASE; FUNCTIONAL RECOVERY; BRAIN ISCHEMIA; CELL-LINE; OCCLUSION; DAMAGE; CORTEX; TRANSPLANTATION; GLUTAMATE AB The efficacy of hyperbaric oxygen (HBO) therapy for treatment of stroke remains to be validated in the laboratory. We report here that adult rats subjected to occlusion of the middle cerebral artery and subsequently exposed to HBO (3 atm, 2 x 90 min at a 24-h intervals; animals terminated shortly after the second treatment) or hyperbaric pressure (HBP; 3 atm, 2 x 90 min at a 24-h interval; animals terminated shortly after the second treatment) immediately after the ischemia or after a 60-min delay generally displayed recovery from motor deficits at 2.5 and 24 h of reperfusion, as well as a reduction in cerebral infarction at 24 h of reperfusion compared to ischemic animals subjected to normal atmospheric pressure. While both HBO and HBP treatments promoted beneficial effects, HBO produced more consistent protection than HBP. Treatment with HBO immediately or 60 min after reperfusion equally produced significant attenuations of cerebral infarction and motor deficits. In contrast, protective effects of HBP treatment against ischemia were noted only when administered immediately after ischemia, which resulted in a significantly reduced infarction volume, but only produced a trend toward decreased behavioral deficits. The present results demonstrate that HBO and, to some extent, HBP reduced ischemic brain damage and behavioral dysfunctions. C1 NIDA, Intramural Res Program, NIH, Baltimore, MD 21224 USA. Natl Def Med Ctr, Grad Inst Med Sci, Taipei, Taiwan. Chi Mei Hosp, Tainan, Taiwan. RP Borlongan, CV (reprint author), NIDA, Intramural Res Program, NIH, Baltimore, MD 21224 USA. OI Borlongan, Cesar/0000-0002-2966-9782 NR 55 TC 50 Z9 57 U1 1 U2 2 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0014-4886 J9 EXP NEUROL JI Exp. Neurol. PD DEC PY 2000 VL 166 IS 2 BP 298 EP 306 DI 10.1006/exnr.2000.7506 PG 9 WC Neurosciences SC Neurosciences & Neurology GA 380PB UT WOS:000165709300010 PM 11085895 ER PT J AU Lee, J Herman, JP Mattson, MP AF Lee, J Herman, JP Mattson, MP TI Dietary restriction selectively decreases glucocorticoid receptor expression in the hippocampus and cerebral cortex of rats SO EXPERIMENTAL NEUROLOGY LA English DT Article DE aging; Alzheimer's disease; caloric restriction; corticosterone; mineralocorticoid ID MESSENGER-RNA EXPRESSION; CHRONIC FOOD RESTRICTION; CALORIC RESTRICTION; GENE-EXPRESSION; OXIDATIVE INJURY; STRESS PROTEINS; BRAIN-DAMAGE; AGED RATS; CORTICOSTERONE; NEURONS AB Dietary restriction (DR) can extend life span and reduce the incidence of age related disease in rodents and primates. DR can be considered as a metabolic stress and might therefore be expected to modify neuroendocrine systems that regulate stress responses. We now report that maintenance of adult rats on a DR regimen results in a significant decrease in the levels of glucocorticoid receptor mRNA and protein in the hippocampus and cerebral cortex, without a change in levels of mineralocorticoid receptors. These findings suggest that DR can alter the responsiveness of brain cells to glucocorticoids, an adaptation that may contribute to beneficial effects of DR on neuronal plasticity and survival demonstrated in recent studies. (C) 2000 Academic Press. C1 NIA, Neurosci Lab, Baltimore, MD 21224 USA. Univ Kentucky, Dept Anat & Neurobiol, Lexington, KY 40536 USA. Univ Kentucky, Sanders Brown Ctr Aging, Lexington, KY 40536 USA. Univ Cincinnati, Med Ctr, Dept Psychiat, Cincinnati, OH 45267 USA. RP Lee, J (reprint author), NIA, Neurosci Lab, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012; Lee, Jaewon/N-9064-2013; Herman, James/D-4960-2015 OI Herman, James/0000-0003-3571-2406 NR 53 TC 37 Z9 38 U1 1 U2 3 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0014-4886 J9 EXP NEUROL JI Exp. Neurol. PD DEC PY 2000 VL 166 IS 2 BP 435 EP 441 DI 10.1006/exnr.2000.7512 PG 7 WC Neurosciences SC Neurosciences & Neurology GA 380PB UT WOS:000165709300023 PM 11085908 ER PT J AU Cragg, GM Newman, DJ AF Cragg, GM Newman, DJ TI Antineoplastic agents from natural sources: achievements and future directions SO EXPERT OPINION ON INVESTIGATIONAL DRUGS LA English DT Review DE cancer chemotherapy; natural products; pharmacophores ID NATIONAL-CANCER-INSTITUTE; DRUG DISCOVERY; POLYKETIDE SYNTHASES; ANTICANCER DRUG; PRODUCTS; ANGIOGENESIS; ECTEINASCIDIN-743; ELEUTHEROBIN; DOXORUBICIN; COMMUNITIES AB The influence of natural products upon anticancer drug discovery and design cannot be overestimated. Approximately 60% of all drugs now in clinical trials for the multiplicity of cancers are either natural products, compounds derived from natural products, contain pharmacophores derived from active natural products or are 'old drugs in new clothes', where (modified) natural products are attached to targeting systems. This review covers those materials that the authors are aware of as being in clinical trials through early 2000 and demonstrates how, even today, in the presence of massive numbers of agents from combinatorial libraries, the compounds produced by 'Mother Nature' are still in the forefront of cancer chemotherapeutics as sources of active chemotypes. C1 NCI, Nat Prod Branch, Dev Therapeut Program, Div Canc Treatment & Diag,Frederick Fairview Ctr, Frederick, MD 21702 USA. RP Cragg, GM (reprint author), NCI, Nat Prod Branch, Dev Therapeut Program, Div Canc Treatment & Diag,Frederick Fairview Ctr, POB B, Frederick, MD 21702 USA. EM cragg@dtpax2.ncifcrf.gov NR 88 TC 52 Z9 56 U1 0 U2 6 PU INFORMA HEALTHCARE PI LONDON PA TELEPHONE HOUSE, 69-77 PAUL STREET, LONDON EC2A 4LQ, ENGLAND SN 1354-3784 J9 EXPERT OPIN INV DRUG JI Expert Opin. Investig. Drugs PD DEC PY 2000 VL 9 IS 12 BP 2783 EP 2797 DI 10.1517/13543784.9.12.2783 PG 15 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 386UW UT WOS:000166080800003 PM 11093353 ER PT J AU Turpin, JA Howard, OMZ AF Turpin, JA Howard, OMZ TI Inhibitors of HIV cellular fusion SO EXPERT OPINION ON THERAPEUTIC PATENTS LA English DT Review DE AIDS; chemokine receptors; fusion inhibitors; gp41; HIV ID HUMAN-IMMUNODEFICIENCY-VIRUS; CHEMOKINE RECEPTOR CXCR4; IN-VIVO; SMALL-MOLECULE; ANTI-HIV-1 ACTIVITY; CCR5 ANTAGONIST; HIGHLY POTENT; INFECTION; TYPE-1; REPLICATION AB HIV infection continues to be a major global health problem. Current anti-HIV therapies targeting reverse transcriptase and protease enzymes suffer from high cost, a high probability of engendering resistance and adverse side effects following prolonged use. Thus, we are faced with the need to develop new antiviral strategies with more potent compounds and/or novel antiviral targets. The recent characterisation of the HIV cell-fusion mechanism and initial mapping of the interactions of the proteins involved in this process has provided an opportunity to identify and take advantage of chemokine co-receptors as new antiviral targets. The HIV fusogenic particle consists of virus-derived gp120, gp41 and cell-derived CD4 and chemokine co-receptors, all of which must interact in a concerted fashion to allow entry of the virus into the cell. The structural analysis of these components has resulted in the identification of a number of new antiviral fusion targets that are distinct from gp120:CD4 binding. Three types of fusogenic particle antagonists have emerged: (1) ribozyme based gene therapy targeting the chemokine co-receptors; (2) peptide-based antagonists targeting either domains of gp41 or chemokine co-receptors; and (3) small molecule inhibitors targeting the virus:co-receptor interaction. In summary, HIV fusion inhibitors, like the current clinically approved agents, will need to be used in combinations consisting of antivirals that target all aspects of the HIV replication cycle and components of the fusogenic particle, to obtain optimum therapeutic effect. C1 NCI, Mol Immunoregulat Lab, Div Basic Sci, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. So Res Inst, Infect Dis Res Dept, Frederick, MD 21701 USA. RP Howard, OMZ (reprint author), NCI, Mol Immunoregulat Lab, Div Basic Sci, Frederick Canc Res & Dev Ctr, Bld 560,Rm 31 19, Frederick, MD 21702 USA. RI Howard, O M Zack/B-6117-2012 OI Howard, O M Zack/0000-0002-0505-7052 NR 49 TC 4 Z9 4 U1 2 U2 3 PU ASHLEY PUBLICATIONS LTD PI LONDON PA UNITEC HOUSE, 3RD FL, 2 ALBERT PLACE FINCHLEY CENTRAL, LONDON N3 1QB, ENGLAND SN 1354-3776 J9 EXPERT OPIN THER PAT JI Expert Opin. Ther. Patents PD DEC PY 2000 VL 10 IS 12 BP 1899 EP 1909 DI 10.1517/13543776.10.12.1899 PG 11 WC Chemistry, Medicinal; Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 378YX UT WOS:000165614700007 ER PT J AU Eiden, LE AF Eiden, LE TI The vesicular neurotransmitter transporters: current perspectives and future prospects SO FASEB JOURNAL LA English DT Review ID CHOLINERGIC GENE LOCUS; CAENORHABDITIS-ELEGANS; ACETYLCHOLINE TRANSPORTER; MONOAMINE TRANSPORTER; CHROMAFFIN GRANULES; SYNAPTIC VESICLES; MOLECULAR-BIOLOGY; RELEASE; STORAGE; GABA C1 NIMH, Mol Neurosci Sect, Lab Cellular & Mol Regulat, Bethesda, MD 20892 USA. RP Eiden, LE (reprint author), NIMH, Mol Neurosci Sect, Lab Cellular & Mol Regulat, 9000 Rockville Pike,Bldg 36,Rm 2A-11, Bethesda, MD 20892 USA. OI Eiden, Lee/0000-0001-7524-944X NR 45 TC 52 Z9 53 U1 0 U2 0 PU FEDERATION AMER SOC EXP BIOL PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0892-6638 J9 FASEB J JI Faseb J. PD DEC PY 2000 VL 14 IS 15 BP 2396 EP 2400 DI 10.1096/fj.00-0817rev PG 5 WC Biochemistry & Molecular Biology; Biology; Cell Biology SC Biochemistry & Molecular Biology; Life Sciences & Biomedicine - Other Topics; Cell Biology GA 380VD UT WOS:000165723400009 PM 11099457 ER PT J AU Weihe, E Eiden, LE AF Weihe, E Eiden, LE TI Chemical neuroanatomy of the vesicular amine transporters SO FASEB JOURNAL LA English DT Review DE aminergic neuron; endocrine cell; CNS; amine-handling cell; autonomic; immune; inflammatory ID CENTRAL-NERVOUS-SYSTEM; CHOLINERGIC GENE LOCUS; ACETYLCHOLINE TRANSPORTER; MONOAMINE TRANSPORTER; ENDOCRINE-CELLS; NEUROTRANSMITTER PHENOTYPE; TYROSINE-HYDROXYLASE; ALZHEIMERS-DISEASE; SYNAPTIC VESICLES; MESSENGER-RNAS AB Acetylcholine, catecholamines, serotonin, and histamine are classical neurotransmitters, These small molecules also play important roles in the endocrine and immune/inflammatory systems, Serotonin secreted from enterochromaffin cells of the gut epithelium regulates gut motility; histamine secreted from basophils and mast cells is a major regulator of vascular permeability and skin inflammatory responses; epinephrine is a classical hormone released from the adrenal medulla, Each of these molecules is released from neural, endocrine, or immune/inflammatory cells only in response to specific physiological stimuli. Regulated secretion is possible because amines are stored in secretory vesicles and released via a stimulus-dependent exocytotic event. Amine storage-at concentrations orders of magnitude higher than in the cytoplasm-is accomplished in turn by specific secretory vesicle transporters that recognize the amines and move them from the cytosol into the vesicle. Immunohistochemical visualization of specific vesicular amine transporters (VATs) in neuronal, endocrine, and inflammatory cells provides important new information about how amine-handling cell phenotypes arise during development and how vesicular transport is regulated during homeostatic response events, Comparison of the chemical neuroanatomy of VATs and amine biosynthetic enzymes has also revealed cell groups that express vesicular transporters but not enzymes for monoamine synthesis, and vice versa: their function and regulation is a new topic of investigation in mammalian neurobiology, The chemical neuroanatomy of the vesicular amine transporters is reviewed here. These and similar data emerging from the study of the localization of the recently characterized vesicular inhibitory and excitatory amino acid transporters will contribute to understanding chemically coded synaptic circuitry in the brain, and amine-handling neuroendocrine and immune/inflammatory cell regulation. C1 Univ Marburg, Dept Mol Neuroimmunol, Inst Anat & Cell Biol, D-35033 Marburg, Germany. NIMH, Mol Neurosci Sect, Lab Cellular & Mol Regulat, Bethesda, MD 20892 USA. RP Weihe, E (reprint author), Univ Marburg, Dept Mol Neuroimmunol, Inst Anat & Cell Biol, Robt Koch Str 6, D-35033 Marburg, Germany. OI Eiden, Lee/0000-0001-7524-944X NR 74 TC 100 Z9 101 U1 2 U2 6 PU FEDERATION AMER SOC EXP BIOL PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0892-6638 J9 FASEB J JI Faseb J. PD DEC PY 2000 VL 14 IS 15 BP 2435 EP 2449 DI 10.1096/fj.00-0202rev PG 15 WC Biochemistry & Molecular Biology; Biology; Cell Biology SC Biochemistry & Molecular Biology; Life Sciences & Biomedicine - Other Topics; Cell Biology GA 380VD UT WOS:000165723400013 PM 11099461 ER PT J AU Uhl, GR Li, S Takahashi, N Itokawa, K Lin, ZC Hazama, M Sora, I AF Uhl, GR Li, S Takahashi, N Itokawa, K Lin, ZC Hazama, M Sora, I TI The VMAT2 gene in mice and humans: amphetamine responses, locomotion, cardiac arrhythmias, aging, and vulnerability to dopaminergic toxins SO FASEB JOURNAL LA English DT Review DE VMAT2; serotonin; histamine; dopamine; norepinephrine; synaptic vesicle; Parkinsonism; MPTP ID VESICULAR MONOAMINE TRANSPORTER-2; CHROMAFFIN GRANULES; MOLECULAR-CLONING; KNOCKOUT MICE; COCAINE; ADDICTION; TOXICITY; DEATH AB Monoamine compartmentalization in monoaminergic neurons uses serial action of the plasma membrane and vesicular monoamine (VAMT2) transporters. We can now define the sequences of the genes encoding these transporters in mice and humans, examine influences of deletions of this gene and alteration in its expression levels in transgenic mice, and identify sequence polymorphisms in the human VMAT2 gene. Examination of VMAT2 variants can provide potential insights into roles for allelic variants at these loci in variant drug responses and in diseases linked to monoaminergic systems, including substance abuse and Parkinson's disease. C1 NIDA, Mol Neurobiol Branch, IRP, NIH, Baltimore, MD 21224 USA. RP Uhl, GR (reprint author), NIDA, Mol Neurobiol Branch, IRP, NIH, Rm 302,5500 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 20 TC 35 Z9 36 U1 0 U2 4 PU FEDERATION AMER SOC EXP BIOL PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0892-6638 J9 FASEB J JI Faseb J. PD DEC PY 2000 VL 14 IS 15 BP 2459 EP 2465 DI 10.1096/fj.00-0205rev PG 7 WC Biochemistry & Molecular Biology; Biology; Cell Biology SC Biochemistry & Molecular Biology; Life Sciences & Biomedicine - Other Topics; Cell Biology GA 380VD UT WOS:000165723400015 PM 11099463 ER PT J AU Paul, RK Jabbar, A Rashid, MA AF Paul, RK Jabbar, A Rashid, MA TI Antiulcer activity of Mikania cordata SO FITOTERAPIA LA English DT Article DE Mikania cordata; alkaloids; antiulcer activity ID GERMACRANOLIDES; DILACTONES AB The alkaloidal fraction obtained from an ethanolic extract of the leaves of Mikania cordata exhibited significant in vivo antiulcer activity in diclofenac sodium-induced gastric erosions in Long Evans rats. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Univ Dhaka, Fac Pharm, Phytochem Res Lab, Dhaka 1000, Bangladesh. RP Rashid, MA (reprint author), NCI, SAIC Frederick, Frederick Canc Res & Dev Ctr, Bldg 560,Rm 32-63B,POB B, Frederick, MD 21702 USA. NR 13 TC 11 Z9 12 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0367-326X J9 FITOTERAPIA JI Fitoterapia PD DEC PY 2000 VL 71 IS 6 BP 701 EP 703 DI 10.1016/S0367-326X(00)00216-1 PG 3 WC Chemistry, Medicinal; Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 379NM UT WOS:000165649100015 PM 11077180 ER PT J AU Arrington, ED Caldwell, MC Kumaravel, TS Lohani, A Joshi, A Evans, MK Chen, HT Nussenzweig, A Holbrook, NJ Gorospe, M AF Arrington, ED Caldwell, MC Kumaravel, TS Lohani, A Joshi, A Evans, MK Chen, HT Nussenzweig, A Holbrook, NJ Gorospe, M TI Enhanced sensitivity and long-term G2 arrest in hydrogen peroxide-treated Ku80-Null cells are unrelated to DNA repair defects SO FREE RADICAL BIOLOGY AND MEDICINE LA English DT Article DE Ku80; hydrogen peroxide; FACS; gamma irradiation; comet assay; cytotoxicity; free radicals ID DEPENDENT PROTEIN-KINASE; HUMAN KU AUTOANTIGEN; STRAND BREAK REPAIR; RNA-POLYMERASE-I; TOPOISOMERASE-II; OXIDATIVE STRESS; DAMAGE; CYCLE; GADD45; EXPRESSION AB While the Ku complex, comprised of Ku70 and Ku80, is primarily involved in the repair of DNA double-strand breaks, it is also believed to participate in additional cellular processes. Here, treatment of embryo fibroblasts (MEFs) derived from either wild-type or Ku80-null (Ku80(-/-)) mice with various stress agents revealed that hydrogen peroxide (H(2)O(2)) was markedly more cytotoxic for Ku80(-/-) MEFs and led to their long-term accumulation in the G2 phase. This differential response was not due to differences in DNA repair, since H(2)O(2)-triggered DNA damage was repaired with comparable efficiency in both Wt and Ku80-/- MEFs, but was associated with differences in the expression of important cell cycle regulatory genes. Our results support the notion that Ku80-mediated cytoprotection and G2-progression are not only dependent on the cell's DNA repair but also may reflect Ku80's influence on additional cellular processes such as gene expression. (C) 2000 Elsevier Science Inc. C1 NIA, Biol Chem Lab, NIH, Baltimore, MD 21224 USA. NIA, Mol Genet Lab, NIH, Baltimore, MD 21224 USA. NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. RP Gorospe, M (reprint author), NIA, Biol Chem Lab, NIH, Box 12,5600 Nathan Shock Dr, Baltimore, MD 21224 USA. EM myriam-gorospe@nih.gov NR 46 TC 23 Z9 24 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0891-5849 J9 FREE RADICAL BIO MED JI Free Radic. Biol. Med. PD DEC 1 PY 2000 VL 29 IS 11 BP 1166 EP 1176 DI 10.1016/S0891-5849(00)00439-1 PG 11 WC Biochemistry & Molecular Biology; Endocrinology & Metabolism SC Biochemistry & Molecular Biology; Endocrinology & Metabolism GA 378RU UT WOS:000165598100015 PM 11121725 ER PT J AU Brant, SR Panhuysen, CIM Bailey-Wilson, JE Rohal, PM Lee, S Mann, J Ravenhill, G Kirschner, BS Hanauer, SB Cho, JH Bayless, TM AF Brant, SR Panhuysen, CIM Bailey-Wilson, JE Rohal, PM Lee, S Mann, J Ravenhill, G Kirschner, BS Hanauer, SB Cho, JH Bayless, TM TI Linkage heterogeneity for the IBD1 locus in Crohn's disease pedigrees by disease onset and severity SO GASTROENTEROLOGY LA English DT Article ID INFLAMMATORY BOWEL-DISEASE; GENOME-WIDE SEARCH; SUSCEPTIBILITY LOCI; CLINICAL CHARACTERISTICS; ALZHEIMERS-DISEASE; PROVIDES EVIDENCE; CHROMOSOME-16; FAMILIES; AGE; CONCORDANCE AB Background & Aims: There is evidence for the IBD1 Crohn's disease'(CD) susceptibility locus on chromosome 16 in several but not all populations studied. Genetic and phenotypic heterogeneity may underlie ability to replicate IBD1. We determined if age and severity stratification could identify a clinical subgroup at risk for IBD1. Methods: Linkage analysis at microsatellites spanning chromosome 16 was performed in 2 groups of CD pedigrees: group 1, 57 pedigrees with at least one affected relative classified as having "severe" disease, by history of surgical resection or immunomodulator therapy, and with disease diagnosed before age 22; and group 2, 33 pedigrees with no history of early-onset, severe CD. Results: Group 1 pedigrees demonstrated genomewide significant linkage evidence for the IBD1 locus (nonparametric multipoint logarithm of the odds [Mlod], 3.84; P = 1.3 x 10(-5)) with linkage evidence greater than all 90 pedigrees (Mlod, 2.12; P = 9.0 x 10(-4)). Group 2 pedigrees had near zero nonparametric 2-point and Mlod scores for the IBD1 region. Heterogeneity between groups 1 and 2 was significant (P = 0.002). Conclusions: Presence of early-onset, more severe CD identifies pedigrees at high risk for IBD1. These pedigrees will have move power to refine the IBD1 locus and identify the causative gene. C1 Johns Hopkins Univ, Sch Med, Harvey M & Lyn P Meyerhoff Inflammatory Bowel Dis, Dept Med, Baltimore, MD 21205 USA. NHGRI, NIH, Baltimore, MD USA. Boston Univ, Sch Med, Genet Program, Dept Med, Boston, MA 02118 USA. Univ Chicago Hosp, Dept Med, Gastroenterol Sect, Martin Boyer Labs, Chicago, IL 60637 USA. RP Brant, SR (reprint author), Johns Hopkins Univ, Sch Med, Harvey M & Lyn P Meyerhoff Inflammatory Bowel Dis, Dept Med, 918 Ross Res Bldg,720 Rutland Ave, Baltimore, MD 21205 USA. OI Bailey-Wilson, Joan/0000-0002-9153-2920 FU NIDDK NIH HHS [K08 DK 02560, P30 DK42086, R01 DK 55731] NR 38 TC 54 Z9 56 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0016-5085 J9 GASTROENTEROLOGY JI Gastroenterology PD DEC PY 2000 VL 119 IS 6 BP 1483 EP 1490 DI 10.1053/gast.2000.20245 PG 8 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 382QM UT WOS:000165833800011 PM 11113069 ER PT J AU Atchison, CR West, AB Balakumaran, A Hargus, SJ Pohl, LR Daiker, DH Aronson, JF Hoffmann, WE Shipp, BK Treinen-Moslen, M AF Atchison, CR West, AB Balakumaran, A Hargus, SJ Pohl, LR Daiker, DH Aronson, JF Hoffmann, WE Shipp, BK Treinen-Moslen, M TI Drug enterocyte adducts: Possible causal factor for diclofenac enteropathy in rats SO GASTROENTEROLOGY LA English DT Article ID NONSTEROIDAL ANTIINFLAMMATORY DRUGS; LIVER PROTEIN ADDUCTS; IMMUNOCHEMICAL DETECTION; SMALL-BOWEL; INDOMETHACIN; COVALENT; INJURY; ACID; GLUCURONIDE; INTESTINE AB Background & Aims: Enteropathy is a frequent complication of diclofenac and other nonsteroidal anti-inflammatory drugs, yet little is known about the underlying mechanism. One possibility is that reactive metabolites of diclofenac form adducts with enterocyte macromolecules, as previously shown for liver. We addressed this possibility by using immunohistochemistry to detect diclofenac adducts. Methods: Rats were treated ovally with diclofenac (10-100 mg/kg) and killed after 1-24 hours, and their gastrointestinal (GI) tracts were evaluated for ulcer number and area. Adduct distribution and intensity were assessed by immunohistochemistry by using a technique to simultaneously process and stain multiple intestinal rings. Results: Drug treatment led to dose-dependent formation of both adducts and ulcers only in small intestine and only in animals with intact enterohepatic circulation. Adducts formed within enterocytes by 1 hour, translocated to the brush border, preceded ulceration and vascular protein leakage, and were intense at sites of ulceration. Adducts and ulcers exhibited a parallel distribution within intestinal quintiles: 3rd > 5th >> 1st. Conclusions: Diclofenac treatment resulted in the formation of drug adducts in enterocytes. Because this molecular change occurred before ulceration, was dose dependent, and exhibited concordant distribution with extent of ulceration, the results suggest a causal role for drug adduct formation in diclofenac enteropathy. C1 Univ Texas, Med Branch, Toxicol Program, Dept Pathol, Galveston, TX 77555 USA. NYU Med Ctr, Dept Pathol, New York, NY USA. 3M Pharmaceut, Pathol & Toxicol, St Paul, MN USA. NHLBI, Lab Mol Immunol, NIH, Bethesda, MD 20892 USA. Univ Illinois, Coll Vet Med, Dept Vet Pathobiol, Urbana, IL 61801 USA. Univ Texas, Med Branch, Dept Prevent Med & Community Hlth, Galveston, TX 77550 USA. RP Treinen-Moslen, M (reprint author), Univ Texas, Med Branch, Toxicol Program, Dept Pathol, Galveston, TX 77555 USA. FU NIDDK NIH HHS [DK 34806]; NIEHS NIH HHS [T32 ES07254] NR 34 TC 47 Z9 49 U1 0 U2 2 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0016-5085 J9 GASTROENTEROLOGY JI Gastroenterology PD DEC PY 2000 VL 119 IS 6 BP 1537 EP 1547 DI 10.1053/gast.2000.20186 PG 11 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 382QM UT WOS:000165833800017 PM 11113075 ER PT J AU Franchimont, D Bouma, G Galon, J Wolkersdorfer, GW Haidan, A Chrousos, GP Bornstein, SR AF Franchimont, D Bouma, G Galon, J Wolkersdorfer, GW Haidan, A Chrousos, GP Bornstein, SR TI Adrenal cortical activation in murine colitis SO GASTROENTEROLOGY LA English DT Article ID INFLAMMATORY BOWEL-DISEASE; TUMOR-NECROSIS-FACTOR; ESTABLISHED EXPERIMENTAL COLITIS; RECOMBINANT INTERLEUKIN-6; LYMPHOCYTIC INFILTRATION; ADRENOCORTICAL TUMORS; IMMUNE-RESPONSES; AXIS ACTIVATION; CELLS; MICE AB Background & Aims: Proper adrenal glucocorticoid secretion is crucial in the course of inflammatory diseases. However, the function and structure of the adrenal glands have not been examined in inflammatory bower diseases. Methods: After induction of trinitrobenzene sulfonic acid (TNBS) colitis in SJL/J mice, plasma hormone and cytokine levels were measured, adrenal structure was analyzed by immunohistochemistry and electron microscopy, and adrenal cytokine/cytokine receptor expression were studied by RNase protection. Results: Adrenals of colitic animals were enlarged and hypervascularized. These animals had a marked increase in plasma corticosterone levels during the course of colitis (270 +/- 34 vs. 16 +/- 11 ng/mL; P < 0.0001) but only a modest elevation of their concurrent adrenocorticotropin levels (57 +/- 13 vs. 29 +/- 9 pmol/L; NS). On electron microscopy, adrenocortical cells showed ultrastructural signs of marked stimulation, and intra-adrenal lymphocytes were frequently found in direct contact with these cells. Concurrent plasma levels of interleukin (IL)-6, the major cytokine activating the hypothalamic-pituitary-adrenal axis, were markedly increased (495 +/- 131 vs. 20 +/- 1.5 pg/mL; P < 0.0001), and this cytokine directly stimulated corticosterone secretion by adrenocortical cells in vitro. Intra-adrenal expression of IL-6 in animals With colitis was increased 80-fold, and the IL-6 receptor subunits Il-6R alpha and gp130 were present in the adrenal cells. Treatment of animals with neutralizing anti-IL-6 antibody reduced the TNBS-induced growth and activation of the adrenal cortices. Conclusions: Colitis is associated with a profound stimulation of adrenocortical cell function and glucocorticoid release. Direct immune-adrenal interactions seem to contribute to this activation of the adrenal glands during colitis. C1 NICHD, Pediat & Reprod Endocrinol Branch, NIH, Bethesda, MD 20892 USA. RP Bornstein, SR (reprint author), NICHD, Pediat & Reprod Endocrinol Branch, NIH, Bldg 10,Room 9D42,10Ctr Dr,MSC 1583, Bethesda, MD 20892 USA. RI bouma, gerd/E-2520-2013 NR 36 TC 32 Z9 33 U1 0 U2 1 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0016-5085 J9 GASTROENTEROLOGY JI Gastroenterology PD DEC PY 2000 VL 119 IS 6 BP 1560 EP 1568 DI 10.1053/gast.2000.20235 PG 9 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 382QM UT WOS:000165833800019 PM 11113077 ER PT J AU Akiyama, Y Watanabe, M Maruyama, K Ruscetti, FW Wiltrout, RH Yamaguchi, K AF Akiyama, Y Watanabe, M Maruyama, K Ruscetti, FW Wiltrout, RH Yamaguchi, K TI Enhancement of antitumor immunity against B16 melanoma tumor using genetically modified dendritic cells to produce cytokines SO GENE THERAPY LA English DT Article DE interleukin (IL)-12; malignant melanoma; genetically modified dendritic cells (DC); intratumoral injection ID RESPONSES IN-VITRO; T-CELLS; INTRATUMORAL INJECTION; CARCINOEMBRYONIC ANTIGEN; IFN-GAMMA; PHASE-I; INTERLEUKIN-12; PEPTIDE; IL-12; VIVO AB Dendritic cells (DC) that have been genetically modified to express cytokine genes may be novel tools for inducing antitumor immune responses. In the present study, the pMX retroviral vector was modified to express the mouse IL-2 (mlL-2pMX) and mouse IL-12 (mlL-12pMX) genes. Supernatants from 293 cells transfected with pMX retroviral vectors were harvested and used to transduce mouse lin bone marrow (BM) progenitor cells. After 48 h co-culture with pseudotype retrovirus, BM cells were cultured for 12 days in the presence of mGM-CSF, mSCF and mTNF-alpha to obtain a DC-enriched fraction. Flow cytometric analysis showed that GFP protein expression in these cultures was 20-40% and that 40-50% of the cultured BM cells were positive for the DC marker, DEC205. About 60% of cells sorted for DEC205 also expressed GFP. The supernatants of DC-mlL-2 and DC-mlL-12 cultured for 48 h contained 5.2 +/- 0.15 and 33.9 +/- 2.6 ng cytokine protein per milliliter, respectively. Intratumoral injection of DC-mlL-2 or DC-mlL-12 on days 8 and 15 after the intradermal injection of 1 x 10(5) B16F10 cells, resulted in a significant reduction in tumor size by day 21, as compared with mice treated with unmodified DC or DC-GFP. Longer term analysis as assessed at day 42 revealed that B16 tumor-bearing mice treated with cytokine gene-modified DC survived significantly longer than mice from other groups. Spleen cells obtained from DC-treated mice were specifically sensitized for the generation of CTL by subsequent restimulation with gene-modified DC. These results suggested that DC genetically modified to express IL-2 or IL-12 can induce potent antitumor responses against well-established, poorly immunogenic B16F10 tumors. C1 Natl Canc Ctr, Res Inst, Div Growth Factor, Chuo Ku, Tokyo 104, Japan. NCI, Expt Therapeut Sect, Expt Immunol Lab, Frederick Canc Res & Dev Ctr, Frederick, MD 21701 USA. NCI, Lab Leukocyte Biol, Frederick Canc Res & Dev Ctr, Frederick, MD 21701 USA. RP Akiyama, Y (reprint author), Natl Canc Ctr, Res Inst, Div Growth Factor, Chuo Ku, 5-1-1 Tsukiji, Tokyo 104, Japan. NR 37 TC 38 Z9 41 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0969-7128 J9 GENE THER JI Gene Ther. PD DEC PY 2000 VL 7 IS 24 BP 2113 EP 2121 DI 10.1038/sj.gt.3301353 PG 9 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity; Research & Experimental Medicine GA 391CD UT WOS:000166335800009 PM 11223993 ER PT J AU Paunu, N Sallinen, SL Karhu, R Miettinen, H Sallinen, P Kononen, J Laippala, P Simola, KOJ Helen, P Haapasalo, H AF Paunu, N Sallinen, SL Karhu, R Miettinen, H Sallinen, P Kononen, J Laippala, P Simola, KOJ Helen, P Haapasalo, H TI Chromosome imbalances in familial gliomas detected by comparative genomic hybridization SO GENES CHROMOSOMES & CANCER LA English DT Article ID SEQUENCE COPY NUMBER; GERM-LINE MUTATIONS; GLIOBLASTOMA-MULTIFORME; TUBEROUS SCLEROSIS; ASTROCYTIC TUMORS; TURCOTS-SYNDROME; P53 MUTATIONS; LINKAGE; LOCUS; GRADE AB Familial occurrence of gliomas, in the absence of well-defined hereditary multisystem disorders, is reported occasionally. We describe 17 families that have been afflicted with two or more gliomas but do not raise suspicion of other inheritable syndromes. The families were identified among 369 consecutive glioma patients operated at the Tampere University Hospital during 1983-1994. We applied comparative genomic hybridization (CGH) analysis on 21 gliomas occurring in these 17 families. The most frequent genetic alterations, detected in over 20% of the tumors, were losses of 6q, 10, 4q, 9p and gains of 7, 19, 20q, Ip. We compared the chromosomal alterations detected in the familial gliomas to those reported previously on 209 sporadic gliomas in nine different CGH studies. In this comparison, the familial gliomas more often showed losses of chromosome arms 4q and 6q and gains of Ip and 22q. The most frequent losses (9/21 tumors) in the familial gliomas resided on chromosome arm 6q (P = 0.005, Fisher's exact test; with Bonferroni correction, P = 0.04). The loss of 6q was also the most common intrafamilial aberration, present in four separate gliomas belonging to two families. The minimal common area of loss on this chromosome resided at 6q14-16. In conclusion, we have found several characteristic aberrations by CGH in the familial gliomas and we present new chromosomal regions possibly involved in the familiar predisposition to gliomas. (C) 2000 Wiley-Liss, Inc. C1 Tampere Univ Hosp, Dept Pathol, FIN-33521 Tampere, Finland. Tampere Univ Hosp, Canc Genet Lab, FIN-33521 Tampere, Finland. NIH, Canc Genet Lab, Natl Ctr Human Genome Res, Bethesda, MD 20892 USA. Univ Tampere, Sch Publ Hlth, FIN-33101 Tampere, Finland. Tampere Univ Hosp, Res Unit, Tampere, Finland. Tampere Univ Hosp, Dept Clin Genet, Tampere, Finland. Tampere Univ Hosp, Dept Neurosurg, Tampere, Finland. RP Paunu, N (reprint author), Tampere Univ Hosp, Dept Pathol, POB 2000, FIN-33521 Tampere, Finland. NR 39 TC 19 Z9 19 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 1045-2257 J9 GENE CHROMOSOME CANC JI Gene Chromosomes Cancer PD DEC PY 2000 VL 29 IS 4 BP 339 EP 346 DI 10.1002/1098-2264(2000)9999:9999<::AID-GCC1049>3.0.CO;2-8 PG 8 WC Oncology; Genetics & Heredity SC Oncology; Genetics & Heredity GA 369JY UT WOS:000165063200008 PM 11066078 ER PT J AU Hunt, KJ Heiss, G Sholinsky, PD Province, MA AF Hunt, KJ Heiss, G Sholinsky, PD Province, MA CA FHS Investigators TI Familial history of metabolic disorders and the Multiple Metabolic Syndrome: The NHLBI Family Heart Study SO GENETIC EPIDEMIOLOGY LA English DT Article DE hypertension; diabetes; dyslipidemia; obesity; family risk score; co-occurrence ID INSULIN-RESISTANCE SYNDROME; MALE TWINS; FASTING INSULIN; GLUCOSE-LEVELS; WOMEN TWINS; HYPERTENSION; DISEASE; POPULATION; OBESITY; PREVALENCE AB A case-control study was conducted to investigate the association between family history of obesity, hypertension, and diabetes and the co-occurrence of metabolic disorders associated with the multiple metabolic syndrome (MMS). Included were 1,448 African and European American men and women aged 48-71 who participated in both the third cohort examination of the Atherosclerosis Risk in Communities study, 1992-1994, and phase I of the Family Heart Study 1993-1995. The joint occurrence of hypertension, dyslipidemia, and diabetes or impaired fasting glucose in an individual determined his/her status of "affected" (MMS: n = 97), while the absence of these three metabolic disorders determined his/her status of "unaffected" (Control: n = 527). First-degree relatives provided the information to calculate family risk scores (FRSs) for the phenotypes under study: obesity, diabetes and hypertension. Although the majority of cases were obese (76.3%), family history of obesity was associated only weakly with the MMS, while family history of diabetes, or hypertension was associated significantly with the MMS (controlling for age, race, gender, and sampling group). Obesity of cases and controls modified the strength of these associations-odds ratios were 2.5 (95% CI:1.1-6.1) and 2.9 (95% CI:1.2-7.0) for the diabetes and hypertension FRSs in the non-obese, while in obese individuals the respective odds ratios were 1.6 (95% CI:0.9-2.8) and 1.7 (95% CI:0.9-3.1). These results may imply that obesity, whether familial or environmental in nature, is associated with the development of the MMS. while in non-obese individuals a family history of diabetes, hypertension, or obesity is a marker of genetic predisposition to components of the MMS. (C) 2000 Wiley-Liss. Inc. C1 Univ N Carolina, CVD Program, Dept Epidemiol, Bank Amer, Chapel Hill, NC 27514 USA. NHLBI, Div Epidemiol & Clin Applicat, Bethesda, MD 20892 USA. Washington Univ, Div Biostat, St Louis, MO USA. RP Hunt, KJ (reprint author), Univ N Carolina, CVD Program, Dept Epidemiol, Bank Amer, 137 E Franklin St,Suite 306, Chapel Hill, NC 27514 USA. FU NHLBI NIH HHS [U01 HL56565, U01 HL56563, U01 HL56564] NR 41 TC 34 Z9 40 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0741-0395 J9 GENET EPIDEMIOL JI Genet. Epidemiol. PD DEC PY 2000 VL 19 IS 4 BP 395 EP 409 DI 10.1002/1098-2272(200012)19:4<395::AID-GEPI10>3.0.CO;2-3 PG 15 WC Genetics & Heredity; Mathematical & Computational Biology SC Genetics & Heredity; Mathematical & Computational Biology GA 376HP UT WOS:000165452400010 PM 11108648 ER PT J AU Vinella, D Cashel, M D'Ari, R AF Vinella, D Cashel, M D'Ari, R TI Selected amplification of the cell division genes ftsQ-ftsA-ftsZ in Escherichia coli SO GENETICS LA English DT Article ID OVERLAPPING TRANSCRIPTIONAL UNITS; PENICILLIN-BINDING PROTEIN-2; GROWTH-RATE CONTROL; GUANOSINE TETRAPHOSPHATE; SALMONELLA-TYPHIMURIUM; RIBOSOMAL-RNA; DNA-SEQUENCE; PPGPP; EXPRESSION; SPOT AB Rapidly growing Escherichia coli is unable to divide in the presence of the antibiotic mecillinam, whose direct target is penicillin-binding protein 2 (PBP2), responsible for the elongation of the cylindrical portion or the cell wall. Division can be restored in the absence of PBP2 activity by increasing the concentration of the cell division proteins FtsQ, FtsA, and FtsZ. We tried to identify regulators of the ftsQ- ftsA-ftsZ operon among mecillinam-resistant mutants, which include strains overexpressing these genes. Dy insertional mutagenesis with mini-Tn10 elements, we selected for insertions that conferred mecillinam resistance. Among 15 such mutants, 7 suppressed the thermosensitivity of the ftsZ84(Ts) mutant, strongly suggesting that they had increased FtsZ activity. In all 7 cases, however, the mutants resulted from a duplication of the ftsQAZ region. These duplications seemed to result from multiple events, suggesting that no simple insertional inactivation can result in a mutant with sufficiently amplified ftsQAZexpression to confer mecillinam resistance. The structure of the duplications suggests a general method for constructing directed duplications of precise sequences. C1 Univ Paris 07, Univ Paris 06, CNRS, Inst Jacques Monod, F-75251 Paris 05, France. NICHHD, Genet Mol Lab, NIH, Bethesda, MD 20892 USA. RP Univ Paris 07, Univ Paris 06, CNRS, Inst Jacques Monod, 2 Pl Jussieu, F-75251 Paris 05, France. EM vinella@ijm.jussieu.fr RI vinella, daniel/G-5504-2012 NR 39 TC 19 Z9 20 U1 0 U2 5 PU GENETICS SOCIETY AMERICA PI BETHESDA PA 9650 ROCKVILLE AVE, BETHESDA, MD 20814 USA SN 0016-6731 EI 1943-2631 J9 GENETICS JI Genetics PD DEC PY 2000 VL 156 IS 4 BP 1483 EP 1492 PG 10 WC Genetics & Heredity SC Genetics & Heredity GA 381AL UT WOS:000165740100004 PM 11102351 ER PT J AU Zhang, J Rosenberg, HF AF Zhang, J Rosenberg, HF TI Sequence variation at two eosinophil-associated ribonuclease loci in humans SO GENETICS LA English DT Article ID SINGLE-NUCLEOTIDE POLYMORPHISMS; RESPIRATORY SYNCYTIAL VIRUS; CATIONIC PROTEIN; GENE FAMILY; ANTIVIRAL ACTIVITY; MOLECULAR-CLONING; INTRONIC ENHANCER; NEUROTOXIN RNS2; RAPID EVOLUTION; SUPERFAMILY AB Host defense against invading pathogens is of great importance to the survival of higher organisms. We hale been studying the evolution of mammalian eosinophil-associated ribonucleases (EARs), which ar-e members of the ribonuclease A super-family with known antipathogen activities. Earlier studies showed that positive: selection promoted rapid diversification of paralogous EAR. genes in both primates and rodents. Intraspecifically, however, it is unknown whether these genes also have divergent alleles. The recent discovery that the gene repertoire of the EAR family is much larger in rodents than in primates has led us to consider the possibility that primates maintain a large number of polymorphic alleles to compensate for a smaller gene repertoire. Here we present sequences of 2417 nucleotides at the two EAR loci, the eosinophil-derived neurotoxin (EDN, RNase 2) and eosinophil cationic protein (ECP, RNase 3), from >50 human individuals. Our data demonstrate dial the nucleotide diversities (0.06-0.11%) at these loci are typical for human nuclear genes, thus permitting us to reject this polymorphism hypothesis. No significant departure from neutrality is noted and no signs of overdominant selection are observed. Similar patterns were observed in a preliminary study of chimpanzees. In summary, our results suggest that the antipathogen functions of the primate EARs are conserved after they are established and that these proteins are not currently undergoing rapid diversification in response to challenge from invading microorganisms. C1 NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. RP Rosenberg, HF (reprint author), NIAID, Host Def Lab, NIH, Bldg 10,Rm 11N104,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 48 TC 45 Z9 49 U1 0 U2 1 PU GENETICS PI BALTIMORE PA 428 EAST PRESTON ST, BALTIMORE, MD 21202 USA SN 0016-6731 J9 GENETICS JI Genetics PD DEC PY 2000 VL 156 IS 4 BP 1949 EP 1958 PG 10 WC Genetics & Heredity SC Genetics & Heredity GA 381AL UT WOS:000165740100039 PM 11102386 ER PT J AU Haoudi, A Mason, JM AF Haoudi, A Mason, JM TI Reverse transcriptase can stabilize or destabilize the genome SO GENOME LA English DT Article DE telomeres; telomerase; retrotransposons; reverse transcriptase ID HUMAN TELOMERIC PROTEIN; DOUBLE-STRAND BREAKS; TRANSPOSON HET-A; DROSOPHILA-MELANOGASTER; CHROMOSOME ENDS; HUMAN-CELLS; SACCHAROMYCES-CEREVISIAE; HYBRID DYSGENESIS; MOLECULAR-BASIS; IMMORTAL CELLS AB Telomeres, the eukaryotic chromosome termini, are deoxyribonucleoprotein structures that distinguish natural chromosome ends from broken DNA. In most organisms, telomeres are extended by a reverse transcriptase (RT) with an integrated RNA template, telomerase; in Drosophila melanogaster, however, telomere-specific retrotransposons, HeT-A and TART, transpose specifically to chromosome ends. Whether telomeres are extended by a telomerase or by retrotransposons, an RT is a key component. RT has been studied extensively, both for its important role in converting RNA genomes to DNA, which has great evolutionary impact, and as a therapeutic target in human retroviral diseases. Here we discuss a few important aspects of RT usage during retrotransposition and telomere elongation. C1 NIEHS, Mol Genet Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Mason, JM (reprint author), NIEHS, Mol Genet Lab, NIH, Res Triangle Pk, NC 27709 USA. NR 97 TC 4 Z9 4 U1 0 U2 0 PU NATL RESEARCH COUNCIL CANADA PI OTTAWA PA RESEARCH JOURNALS, MONTREAL RD, OTTAWA, ONTARIO K1A 0R6, CANADA SN 0831-2796 J9 GENOME JI Genome PD DEC PY 2000 VL 43 IS 6 BP 949 EP 956 DI 10.1139/gen-43-6-949 PG 8 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA 380BC UT WOS:000165677600005 PM 11195348 ER PT J AU Andrews, J Bouffard, GG Cheadle, C Lu, JN Becker, KG Oliver, B AF Andrews, J Bouffard, GG Cheadle, C Lu, JN Becker, KG Oliver, B TI Gene discovery using computational and microarray analysis of transcription in the Drosophila melanogaster testis SO GENOME RESEARCH LA English DT Article ID EXPRESSED SEQUENCE TAGS; PROTEIN GENE; SPERMATOGENESIS; IDENTIFICATION; GENERATION; DATABASE; ENCODES; CLUSTER; GENOME AB Identification and annotation of all the genes in the sequenced Drosophila genome is a work in progress. Wild-type testis function requires many genes and is thus of potentially high value for the identification of transcription units. We therefore undertook a survey of the repertoire of genes expressed in the Drosophila testis by computational and microarray analysis. We generated 3141 high-quality testis expressed sequence tags (ESTs). Testis ESTs computationally collapsed into 1560 cDNA set used for further analysis. OF those, 11% correspond to named genes, and 33% provide biological evidence for a predicted gene. A surprising 47% fail to align with existing ESTs and 16% with predicted genes in the current genome release. EST Frequency and microarray expression profiles indicate that the testis mRNA population is highly complex and shows an extended range of transcript abundance. Furthermore, >80% of the genes expressed in the testis showed onefold overexpression relative to ovaries, or gonadectomized flies. Additionally, >3% showed more than threefold overexpression at p <0.05. Surprisingly, 22% of the genes most highly overexpressed in testis match Drosophila genomic sequence, but not predicted genes. These data strongly support the idea that sequencing additional cDNA libraries From defined tissues, such as testis, will be important tools For refined annotation of the Drosophila genome. Additionally, these data suggest that the number of genes in Drosophila will significantly exceed the conservative estimate of 13,601. C1 NIDDKD, Cellular & Dev Biol Lab, NIH, Bethesda, MD 20892 USA. NHGRI, Bioinformat Grp, Intramural Sequencing Ctr, NIH, Gaithersburg, MD 20877 USA. NIA, DNA Array Unit, NIH, Baltimore, MD 21224 USA. RP Oliver, B (reprint author), NIDDKD, Cellular & Dev Biol Lab, NIH, Bethesda, MD 20892 USA. OI Becker, Kevin/0000-0002-6794-6656 NR 48 TC 151 Z9 151 U1 0 U2 4 PU COLD SPRING HARBOR LAB PRESS PI PLAINVIEW PA 1 BUNGTOWN RD, PLAINVIEW, NY 11724 USA SN 1088-9051 J9 GENOME RES JI Genome Res. PD DEC PY 2000 VL 10 IS 12 BP 2030 EP 2043 DI 10.1101/gr.10.12.2030 PG 14 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 382QN UT WOS:000165833900019 PM 11116097 ER PT J AU Tifft, CJ Proia, RL AF Tifft, CJ Proia, RL TI Stemming the tide: glycosphingolipid synthesis inhibitors as therapy for storage diseases SO GLYCOBIOLOGY LA English DT Review DE glycolipid function; lysosomal storage diseases; neurodegenerative diseases; sphingolipids; substrate deprivation ID PYRAMIDAL NEURON DENDRITOGENESIS; MYELIN-ASSOCIATED GLYCOPROTEIN; GLUCOSYLCERAMIDE SYNTHASE; N-BUTYLDEOXYNOJIRIMYCIN; COMPLEX GANGLIOSIDES; LYSOSOMAL STORAGE; GM2 GANGLIOSIDE; TAY-SACHS; GLYCOLIPID BIOSYNTHESIS; GAUCHER-DISEASE AB Glycosphingolipids (GSLs) are plasma membrane components of every eukaryotic cell. They are composed of a hydrophobic ceramide moiety linked to a glycan chain of variable length and structure, Once thought to be relatively inert, GSLs have now been implicated in a variety of biological processes, Recent studies of animals rendered genetically deficient in various classes of GSLs have demonstrated that these molecules are important for embryonic differentiation and development as well as central nervous system function. A family of extremely severe diseases is caused by inherited defects in the lysosomal degradation pathway of GSLs, In many of these disorders GSLs accumulate in cells, particularly neurons, causing neurodegeneration and a shortened life span. No effective treatment exists for most of these diseases and little is understood about the mechanisms of pathogenesis, This review will discuss the development of a new approach to the treatment of GSL storage disorders that targets the major synthesis pathway of GSLs to stem their cellular accumulation. C1 NIDDKD, Genet Dev & Dis Branch, NIH, Bethesda, MD 20892 USA. Childrens Natl Med Ctr, Dept Med Genet, Washington, DC 20010 USA. RP Proia, RL (reprint author), NIDDKD, Genet Dev & Dis Branch, NIH, Bldg 10,Room 9N314, Bethesda, MD 20892 USA. RI Proia, Richard/A-7908-2012 NR 63 TC 31 Z9 32 U1 1 U2 3 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0959-6658 J9 GLYCOBIOLOGY JI Glycobiology PD DEC PY 2000 VL 10 IS 12 BP 1249 EP 1258 DI 10.1093/glycob/10.12.1249 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 393PV UT WOS:000166479600001 PM 11159916 ER PT J AU Kingsley, PD Ten Hagen, KG Maltby, KM Zara, J Tabak, LA AF Kingsley, PD Ten Hagen, KG Maltby, KM Zara, J Tabak, LA TI Diverse spatial expression patterns of UDP-GalNAc : polypeptide N-acetylgalactosaminyltransferase family member mRNAs during mouse development SO GLYCOBIOLOGY LA English DT Article DE O-glycosylation; mouse development; in situ hybridization; UDP-GalNAc : polypeptide N-acetylgalactosaminyltransferases; epithelial-mesenchymal interactions ID GLYCOCONJUGATE EXPRESSION; LINKED OLIGOSACCHARIDES; EMBRYONIC-DEVELOPMENT; EMBRYOGENESIS; DROSOPHILA; GLYCOPROTEINS; CLONING; COMPLEX; LECTIN; PEANUT AB Cell migration and adhesion during embryonic development are complex processes which likely involve interactions among cell-surface carbohydrates, While considerable work has implicated proteoglycans in a wide range of developmental events, only limited attention has been directed towards understanding the 7role(s) played by the related class of mucin-type O-glycans, The initial step of mammalian mucin-type O-glycosylation is catalyzed by a family of UDP-GalNAc:polypeptide N-acetylgalactosaminyltransferases (ppGaNTases). The spatial expression patterns of the messenger RNAs of seven ppGaNTase family members were investigated from gastrulation through organogenesis stages of mouse development. The seven glycosyltransferases were expressed in unique patterns during embryogenesis. ppGaNTase-T1, -T2, -T4, and -T9 were expressed more ubiquitously than ppGaNTase-T3, -T5, and -T7, Organ systems with discrete accumulation patterns of ppGaNTase family members include the gastrointestinal tract (intestine, liver, stomach, submandibular gland), nervous system (brain, eye), lung, bone, yolk sac, and developing craniofacial region. The pattern in the craniofacial region included differential expression by family members in developing mandible, teeth, tongue and discrete regions of the brain including the pens and migratory, differentiating neurons. Additionally, ppGaNTase-T5 accumulates in a subset of mesenchymal cells at the ventral-most portions of the E12.5 maxilla and mandible underlying the dental lamina, The unique spatiotemporal expression of the different ppGaNTase family members during development suggests unique roles for each of these gene products. C1 Univ Rochester, Aab Inst Biomed Sci, Ctr Oral Biol, Rochester, NY 14642 USA. Univ Rochester, Dept Pediat, Rochester, NY 14642 USA. RP Tabak, LA (reprint author), NIH, NIDCR, 31 Ctr Dr MSC 2290,Bldg 31,Room 2C39, Bethesda, MD 20892 USA. FU NIDCR NIH HHS [DE08108] NR 40 TC 43 Z9 44 U1 0 U2 2 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0959-6658 J9 GLYCOBIOLOGY JI Glycobiology PD DEC PY 2000 VL 10 IS 12 BP 1317 EP 1323 DI 10.1093/glycob/10.12.1317 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 393PV UT WOS:000166479600008 PM 11159923 ER PT J AU Lacey, JV Brinton, LA Mortel, R Berman, ML Wilbanks, GD Twiggs, LB Barrett, RJ AF Lacey, JV Brinton, LA Mortel, R Berman, ML Wilbanks, GD Twiggs, LB Barrett, RJ TI Tubal sterilization and risk of cancer of the endometrium SO GYNECOLOGIC ONCOLOGY LA English DT Article DE endometrial cancer; surgical sterilization; tubal sterilization ID OVARIAN-CANCER; BREAST-CANCER; HYSTERECTOMY; LIGATION; SMOKING AB Objective. Surgical sterilization is a common method of contraception among U.S. women, Most surgical sterilizations are tubal ligations, but few studies have investigated their potential impact on endometrial cancer risk, Methods. A case-control study included 405 women diagnosed with endometrial cancer at 5 U.S. medical centers between 1987 and 1990 and 297 age-, race-, and location-matched controls who were identified by random-digit-dialing. Questionnaires ascertained information on tubal sterilization, and logistic regression models generated odds ratios (ORs) to estimate relative risk. Results. The OR and 95% confidence interval for tubal sterilization, which was reported by 47 cases and 40 controls, was 0.9 (0.6-1.4) before adjustment and 1.4 (0.8-2.4) after adjustment for age, parity, and oral contraceptive use. Age at surgery, years since surgery, or calendar years of surgery were not associated with endometrial cancer, and associations did not vary according to parity or stage of disease at diagnosis. Conclusions. Tubal sterilization is not substantially associated with endometrial cancer. (C) 2000 Academic Press. C1 NCI, Environm Epidemiol Branch, Bethesda, MD 20892 USA. Milton S Hershey Med Ctr, Dept Obstet & Gynecol, Hershey, PA USA. Univ Calif Irvine, Dept Obstet & Gynecol, Irvine, CA 92717 USA. Rush Med Coll, Dept Obstet & Gynecol, Chicago, IL 60612 USA. Univ Minnesota, Sch Med, Dept Obstet & Gynecol, Minneapolis, MN 55455 USA. Wake Forest Univ, Bowman Gray Sch Med, Dept Obstet & Gynecol, Winston Salem, NC 27103 USA. RP Lacey, JV (reprint author), NCI, Environm Epidemiol Branch, Bethesda, MD 20892 USA. RI Brinton, Louise/G-7486-2015 OI Brinton, Louise/0000-0003-3853-8562 NR 20 TC 7 Z9 8 U1 0 U2 1 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0090-8258 J9 GYNECOL ONCOL JI Gynecol. Oncol. PD DEC PY 2000 VL 79 IS 3 BP 482 EP 484 DI 10.1006/gyno.2000.5970 PG 3 WC Oncology; Obstetrics & Gynecology SC Oncology; Obstetrics & Gynecology GA 383ZC UT WOS:000165913700023 PM 11104624 ER PT J AU Kato, T Duffey, DC Ondrey, FG Dong, G Chen, Z Cook, JA Mitchell, JB Van Waes, C AF Kato, T Duffey, DC Ondrey, FG Dong, G Chen, Z Cook, JA Mitchell, JB Van Waes, C TI Cisplatin and radiation sensitivity in human head and neck squamous carcinomas are independently modulated by glutathione and transcription factor NF-kappa B SO HEAD AND NECK-JOURNAL FOR THE SCIENCES AND SPECIALTIES OF THE HEAD AND NECK LA English DT Article DE squamous cell carcinoma; cisplatin; radiation; NF-kappa B; glutathione ID GAMMA-GLUTAMYLCYSTEINE SYNTHETASE; CANCER CELL-LINES; PROINFLAMMATORY CYTOKINE EXPRESSION; ADVANCED LARYNGEAL-CANCER; L-BUTHIONINE SULFOXIMINE; S-TRANSFERASE; INDUCTION CHEMOTHERAPY; INDUCED APOPTOSIS; CONSTITUTIVE ACTIVATION; ORGAN PRESERVATION AB Background. Response to neoadjuvant cisplatin-based chemotherapy has been used to predict overall response to chemoradiation therapy and to select patients with head and neck squamous cell carcinoma (HNSCC) for organ preservation therapy in NCI and VA cooperative group trials. However. differ ent molecular determinants have been reported to contribute to sensitivity of cells to cisplatin and radiation, including glutathione (GSH), and activation of nuclear factor-kappaB (NF-kappaB), a transcription factor that regulates cytoprotective genes. We have reported that NF-kappaB is constitutively activated in HNSCC, but the relationship of NF-kappaB to GSH and to cisplatin and radiation sensitivity in HNSCC is unknown. Methods. We examined human HNSCC lines to define the relationship of cisplatin and radiation sensitivity to intracellular GSH and NF-kappaB and determined whether HNSCC could be sensitized to these modalities by lowering the concentration of glutathione with L-buthionine sulfoximine or inhibiting activation of NF-kappaB by expression of a degradation-resistant mutant inhibitor-kappaB alpha. Results. Cisplatin resistance did not predict radiation resistance in three HNSCC call lines, UM-SCC-9, 11B, and, 38. Resistance to cisplatin correlated with intracellular GSH, and depletion of GSH by treatment with L-BSO sensitized UM-SCC-9 cells to cisplatin but not radiation. conversely, radiation resistance was correlated with activation of NF-kappaB. Expression of a mutant Inhibitor-kappaB after gene transfer inhibited NF-kappaB and sensitized UM-SCC-9 cells to radiation but not cisplatin. Conclusions. GSH and transcription factor NF-alphaB can contribute independently to cisplatin and radiation sensitivity of human HNSCC. These results highlight the need to define molecular determinants of chemotherapy and radiation sensitivity for use in the selection of patients and as novel targets for therapy in future chemoradiation therapy trials for organ preservation in patients with HNSCC. (C) 2000 John Wiley & Sons, Inc. C1 Natl Inst Deafness & Commun Disorders, Tumor Biol Sect, Head & Neck Surg Branch, NIH, Bethesda, MD 20892 USA. NCI, Radiat Biol Branch, NIH, Bethesda, MD 20892 USA. RP Van Waes, C (reprint author), Natl Inst Deafness & Commun Disorders, Tumor Biol Sect, Head & Neck Surg Branch, NIH, Bldg 10,Rm 5D55, Bethesda, MD 20892 USA. FU NIDCD NIH HHS [DC-00016] NR 65 TC 69 Z9 74 U1 0 U2 1 PU JOHN WILEY & SONS INC PI NEW YORK PA 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 1043-3074 J9 HEAD NECK-J SCI SPEC JI Head Neck-J. Sci. Spec. Head Neck PD DEC PY 2000 VL 22 IS 8 BP 748 EP 759 DI 10.1002/1097-0347(200012)22:8<748::AID-HED2>3.0.CO;2-6 PG 12 WC Otorhinolaryngology; Surgery SC Otorhinolaryngology; Surgery GA 375GX UT WOS:000165392100001 PM 11084634 ER PT J AU Yoshiji, H Kuriyama, S Miyamoto, Y Thorgeirsson, UP Gomez, DE Kawata, M Yoshii, J Ikenaka, Y Noguchi, R Tsujinoue, H Nakatani, T Thorgeirsson, SS Fukui, H AF Yoshiji, H Kuriyama, S Miyamoto, Y Thorgeirsson, UP Gomez, DE Kawata, M Yoshii, J Ikenaka, Y Noguchi, R Tsujinoue, H Nakatani, T Thorgeirsson, SS Fukui, H TI Tissue inhibitor of metalloproteinases-1 promotes liver fibrosis development in a transgenic mouse model SO HEPATOLOGY LA English DT Article ID MAMMARY-CARCINOMA CELLS; HUMAN HEPATIC LIPOCYTES; FIBROTIC HUMAN LIVER; MATRIX METALLOPROTEINASES; PROGELATINASE-A; GENE-EXPRESSION; STELLATE CELLS; RNA EXPRESSION; RAT-LIVER; B-CELLS AB Tissue inhibitor of metalloproteinases-l (TIMP-1) has been shown to be increased in liver fibrosis development both in murine experimental models and human samples. However, the direct role of TIMP-1 during liver fibrosis development has not been defined. To address this issue, we developed transgenic mice overexpressing human TIMP-1 (hTIMP-1) in the liver under control of the albumin promoter/enhancer. A model of CCl4-induced hepatic fibrosis was used to assess the extent of fibrosis development in TIMP-1 transgenic (TIMP-Tg) mice and control hybrid (Cont) mice. Without any treatment, overexpression of TIMP-1 itself did not induce liver fibrosis. There were no significant differences of pro-(alpha1)-collagen-I, (alpha2)-collagen-IV, and alpha -smooth muscle actin (alpha -SMA) mRNA expression in the liver between TIMP-Tg and Cent-mice, suggesting that overexpression of TIMP-1 itself did not cause hepatic stellate cell (HSC) activation. After 4-week treatment with CCl4 however, densitometric analysis revealed that TIMP-Tg-mice had a seven-fold increase in liver fibrosis compared with the Cent-mice. The hepatic hydroxyproline content and serum hyaluronic acid were also significantly increased in TIMP-Tg-mice, whereas CCl4-induced liver dysfunction was not altered. An active form of matrix metalloproteinases-2 (MMP-2) level in the liver of TIMP-Tg-mice was decreased relative to that in Cent-mice because of the transgenic TIMP-1. Immunohistochemical analysis revealed that collagen-I and collagen-IV accumulation was markedly increased in the liver of CCl4-treated TIMP-Tg-mice with a pattern similar to that of alpha -SMA positive cells. These results suggest that TIMP-1 does not by itself result in liver fibrosis, but strongly promotes liver fibrosis development. C1 Nara Med Univ, Dept Internal Med 3, Kashihara, Nara 6348522, Japan. NCI, Lab Cellular Carcinogenesis & Promot, Tumor Biol & Carcinogenesis Sect, NIH, Bethesda, MD USA. NCI, Div Basic Sci, Expt Carcinogenesis Lab, NIH, Bethesda, MD USA. Quilmes Natl Univ, Dept Sci & Technol, Mol Oncol Lab, Buenos Aires, DF, Argentina. RP Yoshiji, H (reprint author), Nara Med Univ, Dept Internal Med 3, Shijo Cho, Kashihara, Nara 6348522, Japan. OI Gomez, Daniel E/0000-0002-8629-0787 NR 43 TC 171 Z9 192 U1 0 U2 9 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD DEC PY 2000 VL 32 IS 6 BP 1248 EP 1254 DI 10.1053/jhep.2000.20521 PG 7 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 378AZ UT WOS:000165560300009 PM 11093731 ER PT J AU Kershaw, MH Westwood, JA Zhu, ZP Witte, L Libutti, SK Hwu, P AF Kershaw, MH Westwood, JA Zhu, ZP Witte, L Libutti, SK Hwu, P TI Generation of gene-modified T cells reactive against the angiogenic kinase insert domain-containing receptor (KDR) found on tumor vasculature SO HUMAN GENE THERAPY LA English DT Article ID ENDOTHELIAL GROWTH-FACTOR; TYROSINE KINASE; IN-VITRO; INFILTRATING LYMPHOCYTES; EXPRESSION; INHIBITION; IDENTIFICATION; ANTIBODIES; ANTIGEN; CANCER AB The destruction of newly forming tumor vasculature is a promising approach to inhibit tumor growth. The goal of the present study was to investigate whether human lymphocytes gene modified to express a chimeric receptor specific for the angiogenic endothelial cell receptor, KDR, could react against KDR+ cells. Gene-modified lymphocytes specifically lysed KDR+ cells and secreted cytokines in response to KDR+ target cells including human umbilical vein endothelial cells (HUVECs). Anti-KDR lymphocytes induced HUVECs to secrete the chemokine interleukin 8 and upregulate the adhesion molecules VCAM and E-selectin, which may be important in the recruitment of further immune effector cells to tumor. These KDR-specific lymphocytes may be useful in the adoptive immunotherapy of a broad range of cancers by inducing immune-mediated destruction of tumor neovasculature. C1 NCI, Tumor Immunol Sect, Surg Branch, NIH, Bethesda, MD 20892 USA. ImClone Syst, Dept Mol & Cell Biol, New York, NY 10014 USA. RP Hwu, P (reprint author), NCI, Tumor Immunol Sect, Surg Branch, NIH, Bldg 10,Room 2B42,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 36 TC 24 Z9 24 U1 1 U2 1 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1043-0342 J9 HUM GENE THER JI Hum. Gene Ther. PD DEC PY 2000 VL 11 IS 18 BP 2445 EP 2452 DI 10.1089/10430340050207939 PG 8 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Medicine, Research & Experimental SC Biotechnology & Applied Microbiology; Genetics & Heredity; Research & Experimental Medicine GA 383BU UT WOS:000165863900001 PM 11119416 ER PT J AU Evans, JT Cravens, P Lipsky, PE Garcia, JV AF Evans, JT Cravens, P Lipsky, PE Garcia, JV TI Differentiation and expansion of lentivirus vector-marked dendritic cells derived from human CD34(+) cells SO HUMAN GENE THERAPY LA English DT Article ID CYTOTOXIC T-LYMPHOCYTES; HEMATOPOIETIC STEM-CELLS; MEDIATED GENE-TRANSFER; COLONY-STIMULATING FACTOR; ANTIGEN-PRESENTING CELLS; BONE-MARROW; CORD-BLOOD; IN-VITRO; ANTITUMOR IMMUNITY; PROGENITOR CELLS AB The in vitro genetic manipulation of dendritic cells (DCs) for the expression of foreign proteins or peptides will assist in the development of immunotherapeutic approaches to treat cancer, immunological disorders, and/or infectious diseases. Reports have shown the expansion and differentiation of CD34(+) progenitor cells into mature DCs. In this article we describe the differentiation and expansion of lentivirus vector-marked DCs from umbilical cord blood, bone marrow, and cytokine-mobilized peripheral blood CD34(+) cells in the presence of GM-CSF, TNF-alpha, SCF, Flt-3, and IL-4. Lentivirus-marked DCs expressed high levels of enhanced green fluorescent protein and the characteristic DC surface markers CD1a, CD83, HLA-DR, and CD80. Transduced DCs activated allogeneic CD3(+) T cells as efficiently as control (nontransduced) DCs in mixed lymphocyte reactions. These results demonstrate the potential utility of lentivirus-transduced DCs in future immunotherapy protocols. C1 Univ Texas, SW Med Ctr, Dept Internal Med, Div Infect Dis Y9 206, Dallas, TX 75390 USA. Univ Texas, SW Med Ctr, Dept Internal Med, Harold C Simmons Arthrit Res Ctr, Dallas, TX 75390 USA. NIAMSD, Autoimmun Branch, Bethesda, MD 20892 USA. RP Garcia, JV (reprint author), Univ Texas, SW Med Ctr, Dept Internal Med, Div Infect Dis Y9 206, 532 Harry Hines Blvd, Dallas, TX 75390 USA. FU NCI NIH HHS [5T32CA09082, CA-82055]; NIAID NIH HHS [AI-39416] NR 45 TC 11 Z9 16 U1 0 U2 0 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1043-0342 J9 HUM GENE THER JI Hum. Gene Ther. PD DEC PY 2000 VL 11 IS 18 BP 2483 EP 2492 DI 10.1089/10430340050207975 PG 10 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Medicine, Research & Experimental SC Biotechnology & Applied Microbiology; Genetics & Heredity; Research & Experimental Medicine GA 383BU UT WOS:000165863900005 PM 11119420 ER PT J AU Chute, JP Saini, A Wells, M Clark, W Wu, A St Louis, D Blair, P Harlan, D Kaushal, S AF Chute, JP Saini, A Wells, M Clark, W Wu, A St Louis, D Blair, P Harlan, D Kaushal, S TI Preincubation with endothelial cell monolayers increases gene transfer efficiency into human bone marrow CD34(+) CD38(-) progenitor cells SO HUMAN GENE THERAPY LA English DT Article ID HEMATOPOIETIC STEM-CELLS; EX-VIVO EXPANSION; AMPHOTROPIC RETROVIRUS RECEPTOR; IMMUNE-DEFICIENT MICE; CORD BLOOD-CELLS; IN-VITRO; PERIPHERAL-BLOOD; GROWTH-FACTORS; MESSENGER-RNA; LEUKEMIA-VIRUS AB Retroviral gene transfer studies targeting bone marrow CD34(+) CD38(-) stem cells have been disappointing because of the rarity of these cells, their G(0) cell cycle status, and their low or absent expression of surface retroviral receptors. In this study, we examined whether preincubation of bone marrow CD34(+) CD38(-) stem cells with a hematopoietically supportive porcine microvascular endothelial cell line (PMVECs) could impact the cell cycle status and expression of retroviral receptors in pluripotent CD34(+) CD38(-) cells and the efficiency of gene transfer into these primitive target cells. PMVEC coculture supplemented with GM-CSF + IL-3 + IL-6 + SCF + Flt-3 ligand induced >93% of the CD34(+) CD38(-) population to enter the G(1) or G(2)/S/M phase while increasing this population from 1.4% on day 0 to 6.5% of the total population by day 5. Liquid cultures supplemented with the identical cytokines induced 73% of the CD34(+) CD38(-) population into cell cycle but did not maintain cells with the CD34(+) CD38(-) phenotype over time. We found no significant increase in the levels of AmphoR or GaLVR mRNA in PMVEC-expanded CD34(+) CD38(-) cells after coculture. Despite this, the efficiency of gene transfer using either amphotropic vector (PA317) or GaLV vector (PG13) was significantly greater in PMVEC-expanded CD34(+) CD38(-) cells (11.4 +/- 5.6 and 10.9 +/- 5.2%, respectively) than in either steady state bone marrow CD34(+) CD38(-) cells (0.6 +/- 1.7 and 0.2 +/- 0.6%, respectively; p<0.01 and p<0.01) or liquid culture-expanded CD34(+) CD38(-) cells (1.4 +/- 3.5 and 0.0%, respectively; p<0.01 and p<0.01). Since PMVEC coculture induces a high level of cell cycling in human bone marrow CD34(+) CD38(-) cells and expands hematopoietic cells capable of in vivo repopulation, this system offers potential advantages for application in clinical gene therapy protocols. C1 USN, NIDDK Navy Transplantat & Autoimmun Branch, Stem Cell Biol Lab, Med Res Inst, Bethesda, MD 20889 USA. Uniformed Serv Univ Hlth Sci, Dept Med, Bethesda, MD 20889 USA. RP Chute, JP (reprint author), USN, NIDDK Navy Transplantat & Autoimmun Branch, Stem Cell Biol Lab, Med Res Inst, Bldg 46,Room 2417A,8901 Wisconsin Ave, Bethesda, MD 20889 USA. NR 51 TC 6 Z9 6 U1 0 U2 0 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1043-0342 J9 HUM GENE THER JI Hum. Gene Ther. PD DEC PY 2000 VL 11 IS 18 BP 2515 EP 2528 DI 10.1089/10430340050207993 PG 14 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Medicine, Research & Experimental SC Biotechnology & Applied Microbiology; Genetics & Heredity; Research & Experimental Medicine GA 383BU UT WOS:000165863900007 PM 11119422 ER PT J AU Paniagua, OA Bryant, MB Panza, JA AF Paniagua, OA Bryant, MB Panza, JA TI Transient hypertension directly impairs endothelium-dependent vasodilation of the human microvasculature SO HYPERTENSION LA English DT Article DE endothelium; hypertension, essential; blood vessels ID VASCULAR RELAXATION; ACETYLCHOLINE; CIRCULATION AB Hypertension is associated with decreased endothelium-dependent vasodilation. However, whether endothelial dysfunction is a cause or a consequence of elevated blood pressure is unknown. Therefore, to determine whether hypertension can directly induce endothelial dysfunction, we investigated the effect of increases in intra-arterial pressure on endothelium-dependent vasodilation of the human microvasculature. Small arteries (internal diameter 202+/-75 mum) were isolated from gluteal fat biopsies in 12 healthy normotensive subjects (8 men and 4 women; age, 46+/-10 years). Arteries were cannulated and perfused in chambers oxygenated at 37 degreesC. Endothelium-dependent and -independent responses to acetylcholine (Ach; 10(-9) to 10(-4) mol/L) and sodium nitroprusside (SNP; 10(-9) to 10(-4) mol/L), respectively, were obtained after incubating the vessel with incremental intravascular pressures of 50, 80, and 120 mm Hg for 60 minutes each. The response to Ach was also obtained in different arteries after 3 consecutive incubation periods at 50 mm Hg. Arterial internal diameter was measured directly from amplified digital images. A significant reduction in the vasodilator response to Ach was observed with increases in intravascular pressure (mean vasodilation, 62%, 49%, and 26% at 50, 80, and 120 mm Hg, respectively; P<0.001). In contrast, the response to SNP showed a nonsignificant trend toward greater vasodilation with increases in pressure (mean vasodilation, 40%, 52%, and 57% at 50, 80, and 120 mm Hg, respectively; P=0.10). There was no difference in the consecutive dose-response curves to Ach obtained at the same intravascular pressure (mean vasodilation: 48%, 46%, and 49%; P=0.61). Transient increases in intravascular pressure significantly depress endothelium-dependent vasodilation in human resistance arteries. These findings suggest that elevated blood pressure per se may cause endothelial dysfunction in humans and have implications for the pathophysiology of endothelial dysfunction in hypertension. C1 NHLBI, Cardiol Branch, NIH, Bethesda, MD 20892 USA. RP Panza, JA (reprint author), NHLBI, Cardiol Branch, NIH, 10 Ctr Dr,MSC 1650,Bldg 10,Room 7B-15, Bethesda, MD 20892 USA. NR 17 TC 31 Z9 33 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0194-911X J9 HYPERTENSION JI Hypertension PD DEC PY 2000 VL 36 IS 6 BP 941 EP 944 PG 4 WC Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 384EW UT WOS:000165930700004 PM 11116104 ER PT J AU Rao, FW Keiser, HR O'Connor, DT AF Rao, FW Keiser, HR O'Connor, DT TI Malignant pheochromocytoma - Chromaffin granule transmitters and response to treatment SO HYPERTENSION LA English DT Article DE plasma; chromogranins; norepinephrine; epinephrine ID CHROMOGRANIN-A; GENE-EXPRESSION; PLASMA; STORAGE; RADIOIMMUNOASSAY; DIAGNOSIS; SECRETION; 3,4-DIHYDROXYPHENYLGLYCOL; CATECHOLAMINES; ACTIVATION AB Chromaffin granule transmitters such as chromogranin A and catecholamines have been used in the diagnosis of pheochromocytoma, but the diagnostic and prognostic value of chromogranin A have not been explored in malignant pheochromocytoma. We evaluated these transmitters in patients with pheochromocytoma (n = 27), both benign (n = 13) and malignant (n = 14). Patients with benign pheochromocytoma were studied before and after surgical excision (n = 6), whereas patients with malignant pheochromocytoma were evaluated before and after combination chemotherapy with regular cycles of cyclophosphamide/dacarbazine/vincristine (nonrandomized trial in n = 9). During treatment, patient responses to chemotherapy were divided according to anatomic and clinical criteria: responders (n = 5) versus nonresponders (n = 4). Plasma chromogranin A rose progressively (P < 0.0001) from control subjects (48.0 +/- 3.0 ng/mL) to benign pheochromocytoma (188 +/- 40.5 ng/mL) to malignant pheochromocytoma (2932 +/- 960 ng/mL). Parallel changes were seen for plasma norepinephrine (P < 0.0001), though plasma epinephrine was actually lower in malignant than benign pheochromocytoma (P = 0.0182). In bivariate analyses, chromogranin A, norepinephrine, and epinephrine discriminated between pheochromocytoma and control subjects tall P < 0.0001), whereas in a multivariate analyses, norepinephrine was the best discriminator (P = 0.011). Chromogranin A was significantly different in benign versus malignant pheochromocytoma on both bivariate (P = 0.0003) and multivariate (P = 0.011) analyses. After excision of benign pheochromocytoma, chromogranin A (P = 0.028), norepinephrine (P = 0.047), and epinephrine (P = 0.037) all fell to values near normal. During chemotherapy of malignant :pheochromocytoma (n = 9), plasma chromogranin A (P = 0.047) and norepinephrine (P = 0.02) fell but not epinephrine. In 5 responders to chemotherapy, there were significant declines in chromogranin A (P = 0.03) and norepinephrine (P = 0.03) but not epinephrine; in 4 nonresponders, none of the transmitters changed. Plasma chromogranin A varied longitudinally with tumor response and relapse. We conclude that plasma chromogranin A is an effective tool in the diagnosis of pheochromocytoma, and markedly elevated chromogranin A may point to malignant pheochromocytoma. During chemotherapy of malignant pheochromocytoma, chromogranin A can be used to gauge tumor response and relapse. C1 Univ Calif San Diego, Dept Med, San Diego, CA 92161 USA. Univ Calif San Diego, Ctr Genet Mol, San Diego, CA 92161 USA. NHLBI, Bethesda, MD 20892 USA. VA San Diego Healthcare Syst, San Diego, CA USA. RP O'Connor, DT (reprint author), Univ Calif San Diego, Dept Med, 3350 La Jolla Village Dr, San Diego, CA 92161 USA. NR 32 TC 61 Z9 66 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0194-911X J9 HYPERTENSION JI Hypertension PD DEC PY 2000 VL 36 IS 6 BP 1045 EP 1052 PG 8 WC Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 384EW UT WOS:000165930700023 PM 11116123 ER PT J AU Heim, DA Dunbar, CE AF Heim, DA Dunbar, CE TI Hematopoietic stem cell gene therapy: towards clinically significant gene transfer efficiency SO IMMUNOLOGICAL REVIEWS LA English DT Review ID COLONY-STIMULATING FACTOR; AUTOLOGOUS BONE-MARROW; PERIPHERAL-BLOOD LYMPHOCYTES; LONG-TERM EXPRESSION; GREEN FLUORESCENT PROTEIN; HUMAN ADENOSINE-DEAMINASE; APE LEUKEMIA-VIRUS; EX-VIVO EXPANSION; IN-VIVO; PROGENITOR CELLS AB Early clinical gene therapy and gene marking trials using retroviral vectors to transduce hematopoietic stem cells (HSC) revealed mio major shortcomings of this new treatment modality. One nas insufficient expression or even silencing of the integrated vector sequences, and the second was the low gene transfer efficiency achieved to in vivo repopulating cells. It became clear that neither rodent models nor human in vitro surrogate assays for stem cells were predictive of in rim transgene levels in human target cells. Using the rhesus monkey model we have focused on improving gene transfer efficiency into HSC. Immune rejection of transduced cells has been shown to occur in mature peripheral target cells such as lymphocytes or myocytes. Our studies and those from other investigators suggest that transgenes introduced via HSC induces immunologic tolerance towards the foreign product. In vivo priming of target cells, i.e. mobilization of HSC into the circulation with granulocyte colony stimulating factor and stem cell factor as well as optimization of the in vitro transduction conditions, now allow a stable in vivo gene transfer efficiency of up to 10-15% in both lymphoid and myeloid circulating cells in non-human primates, levels that would be adequate for many clinical applications. C1 NHLBI, Hematol Branch, NIH, Bethesda, MD 20892 USA. RP Dunbar, CE (reprint author), Bldg 10 Room 7C103,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 88 TC 26 Z9 26 U1 0 U2 0 PU MUNKSGAARD INT PUBL LTD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0105-2896 J9 IMMUNOL REV JI Immunol. Rev. PD DEC PY 2000 VL 178 BP 29 EP 38 DI 10.1034/j.1600-065X.2000.17802.x PG 10 WC Immunology SC Immunology GA 394NK UT WOS:000166530000005 PM 11213804 ER PT J AU Notarangelo, LD Giliani, S Mazza, C Mella, P Savoldi, G Rodriguez-Perez, C Mazzolari, E Fiorini, M Duse, M Plebani, A Ugazio, AG Vihinen, M Candotti, F Schumacher, RF AF Notarangelo, LD Giliani, S Mazza, C Mella, P Savoldi, G Rodriguez-Perez, C Mazzolari, E Fiorini, M Duse, M Plebani, A Ugazio, AG Vihinen, M Candotti, F Schumacher, RF TI Of genes and phenotypes: the immunological and molecular spectrum of combined immune deficiency. Defects of the gamma(c)-JAK3 signaling pathway as a model SO IMMUNOLOGICAL REVIEWS LA English DT Review ID SEVERE COMBINED IMMUNODEFICIENCY; RECEPTOR-GAMMA-CHAIN; HEMATOPOIETIC PROGENITOR CELLS; BONE-MARROW TRANSPLANTATION; IN-UTERO TRANSPLANTATION; FLOW CYTOMETRIC ANALYSIS; MICE LACKING JAK3; INTERLEUKIN-2 RECEPTOR; LYMPHOID DEVELOPMENT; T-CELLS AB Cytokines play a major role in lymphoid development. Defects of the common gamma chain (gamma (c)) or of the JAK3 protein in humans have been shown to result in a severe combined immune deficiency (SCID), with a profound defect in T and natural killer (NK)-cell development, whereas B-cell generation is apparently unaffected (T-B+NK- SCID). While extensive molecular and biochemical analysis of these patients has been instrumental in understanding better the biological properties of the gamma (c) and JAK3 protein, an unexpected phenotypic heterogeneity of gamma (c) and JAK3 deficiency has emerged, indicating the need for appropriate and extensive investigations even in patients with atypical presentations. At the same time, characterization of the defects has been instrumental in the development of never therapeutic approaches, from in utero hematopoietic stem cell transplantation to gene therapy. C1 Univ Brescia, Pediat Clin, Ist Med Mol Angelo Nocivelli, Brescia, Italy. Tampere Univ, Hosp Med, FIN-33101 Tampere, Finland. NHGRI, Clin Gene Therapy Branch, NIH, Bethesda, MD 20892 USA. RP Notarangelo, LD (reprint author), Univ Brescia, Spedali Civili, Pediat Clin, Ist Med Mol Angelo Nocivelli, I-25123 Brescia, Italy. RI Plebani, Alessandro/C-8593-2011; Vihinen, Mauno/A-8452-2012; Notarangelo, Luigi/F-9718-2016 OI Vihinen, Mauno/0000-0002-9614-7976; Notarangelo, Luigi/0000-0002-8335-0262 FU Telethon [E.0668] NR 78 TC 56 Z9 58 U1 0 U2 5 PU MUNKSGAARD INT PUBL LTD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0105-2896 J9 IMMUNOL REV JI Immunol. Rev. PD DEC PY 2000 VL 178 BP 39 EP 48 DI 10.1034/j.1600-065X.2000.17812.x PG 10 WC Immunology SC Immunology GA 394NK UT WOS:000166530000006 PM 11213805 ER PT J AU Fowke, KR Kaul, R Rosenthal, KL Oyugi, J Kimani, J Rutherford, WJ Nagelkerke, NJD Ball, TB Bwayo, JJ Simonsen, JN Shearer, GM Plummer, FA AF Fowke, KR Kaul, R Rosenthal, KL Oyugi, J Kimani, J Rutherford, WJ Nagelkerke, NJD Ball, TB Bwayo, JJ Simonsen, JN Shearer, GM Plummer, FA TI HIV-1-specific cellular immune responses among HIV-1-resistant sex workers SO IMMUNOLOGY AND CELL BIOLOGY LA English DT Article DE Africa; AIDS; cellular immunity; cytotoxic T lymphocyte; exposed uninfected; HIV; resistance; T helper cell; vaccine ID IMMUNODEFICIENCY-VIRUS TYPE-1; HIV-SEROPOSITIVE INDIVIDUALS; RECOMBINANT VACCINIA VIRUSES; CD4(+) T-CELLS; DISEASE PROGRESSION; VIRAL-INFECTION; NAIROBI; TRANSMISSION; RECEPTOR; LYMPHOCYTES AB The goal of the present study was to determine whether there were HIV-1 specific cellular immune responses among a subgroup of women within a cohort of Nairobi prostitutes (n = 1800) who, despite their intense sexual exposure to HIV-1, are epidemiologically resistant to HIV-1 infection. Of the 80 women defined to be resistant, 24 were recruited for immunological evaluation. The HIV-1-specific T-helper responses were determined by IL-2 production following stimulation with HIV-1 envelope peptides and soluble gp120. Cytotoxic T lymphocyte responses were determined by lysis of autologous EBV-transformed B cell lines infected with control vaccinia virus or recombinant vaccinia viruses containing the HIV-1 structural genes env, gag and pol. Resistant women had significantly increased HIV-1 specific T-helper responses, as determined by in vitro IL-2 production to HIV-1 envelope peptides and soluble glycoprotein 120, compared with low-risk seronegative and HIV-1-infected controls (P less than or equal to 0.01, Student's t-test). Seven of the 17 (41%) resistant women showed IL-2 stimulation indices greater than or equal to 2.0. HIV-1-specific CTL responses were detected among 15/22 (68.2%) resistant women compared with 0/12 low-risk controls (Chi-squared test, P < 0.001). In the two resistant individuals tested, the CTL activity was mediated by CD8(+) effectors. Many HIV-1-resistant women show evidence of HIV-1-specific T-helper and cytotoxic responses. These data support the suggestion that HIV-1-specific T-cell responses contribute to protection against HIV-1 infection. C1 Univ Manitoba, Dept Med Microbiol, Lab Viral Immunol, Winnipeg, MB R3E 0W3, Canada. McMaster Univ, Dept Pathol, Program Immunol, Hamilton, ON, Canada. Univ Nairobi, Dept Med Microbiol, Nairobi, Kenya. NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. RP Fowke, KR (reprint author), Univ Manitoba, Dept Med Microbiol, Lab Viral Immunol, Room 539,730 William Ave, Winnipeg, MB R3E 0W3, Canada. OI Fowke, Keith/0000-0001-8227-6649 NR 58 TC 84 Z9 84 U1 1 U2 2 PU BLACKWELL SCIENCE ASIA PI CARLTON PA 54 UNIVERSITY ST, P O BOX 378, CARLTON, VICTORIA 3053, AUSTRALIA SN 0818-9641 J9 IMMUNOL CELL BIOL JI Immunol. Cell Biol. PD DEC PY 2000 VL 78 IS 6 BP 586 EP 595 DI 10.1046/j.1440-1711.2000.00944.x PG 10 WC Cell Biology; Immunology SC Cell Biology; Immunology GA 379VD UT WOS:000165662800003 PM 11114968 ER PT J AU Noben-Trauth, N AF Noben-Trauth, N TI Susceptibility to Leishmania major infection in the absence of IL-4 SO IMMUNOLOGY LETTERS LA English DT Article DE Leishmania; IL-4; knockout; Th1/Th2 ID RECEPTOR-GAMMA-CHAIN; CD4+ T-CELLS; INTERFERON-GAMMA; INTERLEUKIN-4 RECEPTOR; MURINE LEISHMANIASIS; FUNCTIONAL COMPONENT; DEFICIENT MICE; LYMPHOKINE; RESISTANCE; CLONING AB Infection with the intracellular parasite Leishmania major is the prototypical model system used to study Th1 and Th2 cytokine responses in vivo. Mouse strains that are resistant to L. major produce high levels of IFN gamma, and Th1 cytokines while susceptible BALB/c mice have elevated levels of IL-4, and Th2-associated cytokines during infection. While antibody neutralization of IL-4 or IFN gamma in vivo alters the disease patterns, infection of mice genetically deficient for IL-4 or IL-4 receptor (IL-4R alpha) yield surprising outcomes. Contrary to the Th1/Th2 paradigm, IL-4 -/- and IL-4R alpha -/- mice remain highly susceptible to L. major parasite substrain LV39. In distinct contrast, another L. major substrain, IR173, the IL-4R alpha -/- mice are highly resistant. These findings indicate a disparity between antibody treatment versus gene deletion, and more generally, challenge the role of IL-4 in promoting susceptibility to L. major. Furthermore, IL-4R alpha -/- mice reveal that the ability of L. major to escape immune clearance depends on the parasite substrain. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIAID, Immunol Lab, NIH, Rockville, MD 20852 USA. RP Noben-Trauth, N (reprint author), NIAID, Immunol Lab, NIH, 12441 Parklawn Dr, Rockville, MD 20852 USA. NR 40 TC 11 Z9 11 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0165-2478 J9 IMMUNOL LETT JI Immunol. Lett. PD DEC 1 PY 2000 VL 75 IS 1 BP 41 EP 44 DI 10.1016/S0165-2478(00)00280-7 PG 4 WC Immunology SC Immunology GA 379QA UT WOS:000165652900008 PM 11163865 ER PT J AU Wang, E Marincola, FM AF Wang, E Marincola, FM TI A natural history of melanoma: serial gene expression analysis SO IMMUNOLOGY TODAY LA English DT Article ID METASTATIC MELANOMA; HUMAN CANCER; T-CELLS; TUMOR; ANTIGENS; PATTERNS; PEPTIDE; IMMUNOTHERAPY; MICROARRAYS; LYMPHOCYTES AB Ena Wang and Francesco Marincola propose a novel strategy whereby tumor-host interactions are studied within the melanoma microwenvironment by serial gene expression analysis. Methodological constraints and ways to circumvent them laue discussed. This approach might improve our understanding of the molecular basis of tumor regression in response to immune manipulation. C1 NCI, Ctr Clin, Surg Branch, NIH, Bethesda, MD 20892 USA. NCI, Ctr Clin, Dept Transfus Med, NIH, Bethesda, MD 20892 USA. RP Wang, E (reprint author), NCI, Ctr Clin, Surg Branch, NIH, Bethesda, MD 20892 USA. NR 29 TC 58 Z9 58 U1 0 U2 2 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 0167-5699 J9 IMMUNOL TODAY JI Immunol. Today PD DEC PY 2000 VL 21 IS 12 BP 619 EP 623 DI 10.1016/S0167-5699(00)01724-2 PG 5 WC Immunology SC Immunology GA 383AB UT WOS:000165858600005 PM 11114422 ER PT J AU Ashwell, JD Vacchio, MS Galon, J AF Ashwell, JD Vacchio, MS Galon, J TI Do glucocorticoids participate in thymocyte development? SO IMMUNOLOGY TODAY LA English DT Letter ID POSITIVE SELECTION; MICE C1 NIH, Lab Immune Cell Biol, Bethesda, MD 20892 USA. NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA. Natl Inst Arthrit & Metab, Lymphocyte Cell Biol Sect, NIH, Bethesda, MD 20892 USA. RP Ashwell, JD (reprint author), NIH, Lab Immune Cell Biol, Bldg 10, Bethesda, MD 20892 USA. NR 9 TC 25 Z9 25 U1 0 U2 2 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 0167-5699 J9 IMMUNOL TODAY JI Immunol. Today PD DEC PY 2000 VL 21 IS 12 BP 644 EP 645 DI 10.1016/S0167-5699(00)01758-8 PG 2 WC Immunology SC Immunology GA 383AB UT WOS:000165858600009 PM 11188796 ER PT J AU Martin, KR Kari, FW Barrett, JC French, JE AF Martin, KR Kari, FW Barrett, JC French, JE TI N-acetyl-L-cysteine simultaneously increases mitogenesis and suppresses apoptosis in mitogen-stimulated B-lymphocytes from p53 haploinsufficient Tg.AC (v-Ha-ras) mice SO IN VITRO & MOLECULAR TOXICOLOGY-A JOURNAL OF BASIC AND APPLIED RESEARCH LA English DT Article ID PROGRAMMED CELL-DEATH; LYMPHOMA CELLS; IMMUNE-SYSTEM; IN-VITRO; T-CELLS; CANCER; ACTIVATION; CHEMOPREVENTION; ANTIOXIDANTS; FIBROBLASTS AB Recent epidemiological evidence suggests that antioxidants may enhance carcinogenesis by promoting cellular proliferation and/or impeding programmed cell death. We examined the effect of N-acetyl-L-cysteine (NAC) on mitogenesis and apoptosis in splenocytes from p53 haploinsufficient Tg.AC (v-Ha-ras) mice. This model contains genetic lesions found frequently in human cancer and is predisposed to develop carcinogen-induced cancer. Splenocytes were incubated with NAC alone ox. with the B- and T-cell-specific mitogens Concanavalin A (Con A) and E. coli lipopolysaccharide (LPS), respectively. Mitogenesis increased 17-fold in mitogen-stimulated cultures and. 10-fold in cultures incubated with NAC alone. Go-incubation with both NAG (1000 mug/mL) and mitogen increased mitogenesis by 33-fold without changing apoptosis rates. Strikingly, incubation with NAC and LPS attenuated LPS-induced apoptosis. Mitogen alone did not affect GSH levels but NAC-induced increases were significantly depleted by co-incubation with mitogen, Furthermore, NAC increased the number of CD45R(+) B cells, but decreased CD3(+) T cells showing enhanced. survival of B cells under these conditions. These results demonstrate concurrent reduced. apoptosis and increased mitogenesis in B lymphocytes that may favor clonal selection of preneoplastic cells. C1 NIEHS, Lab Environm Carcinogenesis & Mutagenesis, Transgen Carcinogenesis Unit, NIH, Res Triangle Pk, NC 27709 USA. NIEHS, Mol Carcinogenesis Lab, NIH, Res Triangle Pk, NC 27709 USA. RP French, JE (reprint author), NIEHS, Lab Environm Carcinogenesis & Mutagenesis, Transgen Carcinogenesis Unit, NIH, POB 12233,111 TW Alexander Dr,MD F1-05, Res Triangle Pk, NC 27709 USA. NR 36 TC 7 Z9 7 U1 1 U2 2 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1097-9336 J9 IN VITRO MOL TOXICOL JI In Vitro Mol. Toxicol.-J. Basic Appl. Res. PD WIN PY 2000 VL 13 IS 4 BP 237 EP 247 PG 11 WC Toxicology SC Toxicology GA 419NP UT WOS:000167955400003 ER PT J AU Lukomski, S Nakashima, K Abdi, I Cipriano, VJ Ireland, RM Reid, SD Adams, GG Musser, JM AF Lukomski, S Nakashima, K Abdi, I Cipriano, VJ Ireland, RM Reid, SD Adams, GG Musser, JM TI Identification and characterization of the scl gene encoding a group a Streptococcus extracellular protein virulence factor with similarity to human collagen SO INFECTION AND IMMUNITY LA English DT Article ID GROUP-A STREPTOCOCCI; HYALURONIC-ACID CAPSULE; FIBRONECTIN-BINDING PROTEIN; PYROGENIC EXOTOXIN-B; CYSTEINE PROTEASE; HUMAN GRANULOCYTES; BACTERIAL SURFACE; EPITHELIAL-CELLS; ESCHERICHIA-COLI; PYOGENES AB Group A Streptococcus (GAS) expresses cell surface proteins that mediate important biological functions such as resistance to phagocytosis, adherence to plasma and extracellular matrix proteins, and degradation of host proteins. An open reading frame encoding a protein of 348 amino acid residues was identified by analysis of the genome sequence available for a serotype M1 strain. The protein has an LPATGE sequence located near the carboxy terminus that matches the consensus sequence (LPXTGX) present in many gram-positive cell wall-anchored molecules. Importantly, the central region of this protein contains 50 contiguous Gly-X-X triplet amino acid motifs characteristic of the structure of human collagen. The structural gene (designated scl for streptococcal collagen-like) was present in all 50 GAS isolates tested, which together express 21 different M protein types and represent the breadth of genomic diversity in the species. DNA sequence analysis of the gene in these 50 isolates found that the number of contiguous Gly-X-X motifs ranged from 14 in serotype M6 isolates to 62 in a serotype M41 organism. M1 and M18 organisms had the identical allele, which indicates very recent horizontal g-ne transfer. The gene was transcribed abundantly in the logarithmic but not stationary phase of growth, a result consistent with the occurrence of a DNA sequence with substantial homology with a consensus Mga binding site immediately upstream of the scl open reading frame. Two isogenic mutant M1 strains created by nonpolar mutagenesis of the scl structural gene were not attenuated for mouse virulence as assessed by intraperitoneal inoculation. In contrast, the isogenic mutant derivative made from the M1 strain representative of the subclone most frequently causing human infections was significantly less virulent when inoculated subcutaneously into mice. In addition, both isogenic mutant strains had significantly reduced adherence to human A549 epithelial cells grown in culture. These studies identify a new extracellular GAS virulence factor that is widely distributed in the species and participates in adherence to host cells and soft tissue pathology. C1 NIAID, Lab Human Bacterial Pahtogenesis, Rocky Mt Labs, NIH, Hamilton, MT 59840 USA. Baylor Coll Med, Dept Pathol, Houston, TX 77030 USA. RP Musser, JM (reprint author), NIAID, Lab Human Bacterial Pahtogenesis, Rocky Mt Labs, NIH, 903 S 4th St, Hamilton, MT 59840 USA. FU NIAID NIH HHS [AI33119] NR 62 TC 109 Z9 112 U1 0 U2 3 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD DEC PY 2000 VL 68 IS 12 BP 6542 EP 6553 DI 10.1128/IAI.68.12.6542-6553.2000 PG 12 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 403AH UT WOS:000167020000005 PM 11083763 ER PT J AU Hisaeda, H Stowers, AW Tsuboi, T Collins, WE Sattabongkot, JS Suwanabun, N Torii, M Kaslow, DC AF Hisaeda, H Stowers, AW Tsuboi, T Collins, WE Sattabongkot, JS Suwanabun, N Torii, M Kaslow, DC TI Antibodies to malaria vaccine candidates Pvs25 and Pvs28 completely block the ability of Plasmodium vivax to infect mosquitoes SO INFECTION AND IMMUNITY LA English DT Article ID MINIMAL VARIATION; TARGET ANTIGEN; SEXUAL STAGE; TRANSMISSION; FALCIPARUM; IMMUNITY; SURFACE; PROTEINS; PFS25; POLYMORPHISM AB Transmission-blocking vaccines are one strategy for controlling malaria, whereby sexual-stage parasites are inhibited from infecting mosquitoes by human antibodies. To evaluate whether the recently cloned Plasmodium vivax proteins Pvs25 and Pvs28 are candidates for a transmission-blocking vaccine, the molecules were expressed in yeast as secreted recombinant proteins. Mice vaccinated with these proteins adsorbed to aluminum hydroxide developed strong antibody responses against the immunogens, although for Pvs28, this response was genetically restricted. Antisera against both recombinant Pvs25 and Pvs28 recognized the corresponding molecules expressed by cultured sexual-stage parasites isolated from patients with P. vivax malaria, The development of malaria parasites in mosquitoes was completely inhibited when these antisera were ingested with the infected blood meal. Pvs25 and Pvs28, expressed in Saccharomyces cerevisiae, are as yet the only fully characterized transmission-blocking vaccine candidates against P. vivax that induce such a potent antiparasite response. C1 NIAID, Malaria Vaccine Dev Unit, Parasit Dis Lab, NIH, Rockville, MD 20852 USA. Univ Tokushima, Sch Med, Dept Parasitol& Immunol, Tokushima 7708503, Japan. Ehime Univ, Sch Med, Dept Mol Parasitol, Shigenobu, Ehime 7910295, Japan. Ctr Dis Control & Prevent, Div Parasit Dis, Chamblee, GA 30341 USA. Ctr Dis Control & Prevent, Anim Resources Branch, Sci Resources Program, Natl Ctr Infect Dis, Chamblee, GA 30341 USA. Armed Forces Res Inst Med Sci, Dept Entomol, Bangkok 10400, Thailand. RP Stowers, AW (reprint author), NIAID, Malaria Vaccine Dev Unit, Parasit Dis Lab, NIH, 12441 Parklawn Dr, Rockville, MD 20852 USA. NR 32 TC 103 Z9 114 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD DEC PY 2000 VL 68 IS 12 BP 6618 EP 6623 DI 10.1128/IAI.68.12.6618-6623.2000 PG 6 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 403AH UT WOS:000167020000015 PM 11083773 ER PT J AU Carroll, JA Cordova, RM Garon, CF AF Carroll, JA Cordova, RM Garon, CF TI Identification of 11 pH-regulated genes in Borrelia burgdorferi localizing to linear plasmids SO INFECTION AND IMMUNITY LA English DT Article ID DECORIN-BINDING-PROTEIN; OUTER SURFACE-PROTEIN; LYME-DISEASE SPIROCHETE; DIFFERENTIAL EXPRESSION; IN-VITRO; MOLECULAR CHARACTERIZATION; MEMBRANE-PROTEINS; IXODES-RICINUS; INFECTION; SEQUENCE AB When Borrelia burgdorferi is transmitted from the tick vector to the mammalian host, the bacterium experiences alterations in its environment, such as changes in temperature and pH. Previously, we observed numerous alterations in the membrane protein profile when B. burgdorferi B31 was grown at pH 7.0 compared to pH 8.0, Here we identify 11 genes localizing to linear plasmids that are up-regulated at pH 7.0 relative to pH 8.0 in vitro. Seven genes (bba03, bba24, bba64, bba66, bbe31, bbj4l/bbi39 [encoding products that are 99% identical], and bbk01) were indirectly identified by proteomic analysis of membrane proteins. Another gene, bba36, was identified by screening a B. burgdorferi B31 genomic library with cross-adsorbed hyperimmune rabbit serum. Two additional genes, bba65 and bba73, were identified by Northern blot analysis. Genes bba64, bba65, bba66, bbj41/bbi39, and bba73 are members of paralogous gene family 54, and bbe31 is a member of the closely related paralogous gene family 60. Gene bba24 is part of a bicistronic operon with bba25 that encodes the well-characterized decorin binding proteins A and B. All 11 genes were transcriptionally regulated, yet the degree of pH regulation varied, with some genes more tightly regulated than others. The regions upstream of these pH-regulated genes appeared to be unrelated, yet many contained dyad repeats ranging from 12 to 25 nucleotides in length that may be involved in the regulation of these genes. C1 NIAID, Rocky Mt Labs, Microscopy Branch, Hamilton, MT 59840 USA. RP Carroll, JA (reprint author), 903 S 4th St, Hamilton, MT 59840 USA. NR 38 TC 76 Z9 76 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD DEC PY 2000 VL 68 IS 12 BP 6677 EP 6684 DI 10.1128/IAI.68.12.6677-6684.2000 PG 8 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 403AH UT WOS:000167020000023 PM 11083781 ER PT J AU Carroll, JA Coleman, SA Smitherman, LS Minnick, MF AF Carroll, JA Coleman, SA Smitherman, LS Minnick, MF TI Hemin-binding surface protein from Bartonella quintana SO INFECTION AND IMMUNITY LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; OUTER-MEMBRANE PROTEIN; CONGO RED BINDING; INFLUENZAE TYPE-B; PORPHYROMONAS-GINGIVALIS; SP-NOV; HAEMOPHILUS-INFLUENZAE; ESCHERICHIA-COLI; SALMONELLA-TYPHIMURIUM; BACILLARY ANGIOMATOSIS AB Bartonella quintana, the agent of trench fever and a cause of endocarditis and bacillary angiomatosis in humans, has the highest reported in vitro hemin requirement for any bacterium. We determined that eight membrane-associated proteins from B. quintana bind hemin and that a similar to 25-kDa protein (HbpA) was the dominant hemin binding protein. Like many outer membrane proteins, HbpA partitions to the detergent phase of a Triton X-114 extract of the cell and is heat modifiable, displaying an apparent molecular mass shift from approximately 25 to 30 kDa when solubilized at 100 degreesC. Immunoblots of purified outer and inner membranes and immunoelectron microscopy with whole cells show that HbpA is strictly located in the outer membrane and surface exposed, respectively. The N-terminal sequence of mature HbpA was determined and used to clone the HbpA-encoding gene (hbpA) from a lambda genomic library. The hbpA gene is 816 bp in length, encoding a predicted immature protein of approximately 29.3 kDa and a mature protein of 27.1 kDa. A Fur box homolog with 53% identity to the Escherichia coli Fur consensus is located upstream of hbpA and may be involved in regulating expression. BLAST searches indicate that the closest homologs to HbpA include the Bartonella henselae phage associated membrane protein, Pap31 (58.4% identity), and the OMP31 porin from Brucella melitensis (31.7% identity). High-stringency Southern blots indicate that all five pathogenic Bartonella spp. possess hbpA homologs. Recombinant HbpA can bind hemin in vitro; however, it does not confer a hemin-binding phenotype upon E. coli. Intact B. quintana treated with purified anti-HbpA Fab fragments show a significant (P < 0.004) dose-dependent decrease in hemin binding relative to controls, suggesting that HbpA plays an active role in hemin acquisition and therefore pathogenesis. HbpA is the first potential virulence determinant characterized from B. quintana. C1 Univ Montana, Div Biol Sci, Missoula, MT 59812 USA. NIAID, Microscopy Branch, Rocky Mt Labs, Hamilton, MT 59840 USA. RP Univ Montana, Div Biol Sci, Missoula, MT 59812 USA. EM minnick@selway.umt.edu FU NIAID NIH HHS [R15 AI045534-01, R15 AI045534] NR 69 TC 44 Z9 46 U1 0 U2 3 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0019-9567 EI 1098-5522 J9 INFECT IMMUN JI Infect. Immun. PD DEC PY 2000 VL 68 IS 12 BP 6750 EP 6757 DI 10.1128/IAI.68.12.6750-6757.2000 PG 8 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 403AH UT WOS:000167020000033 PM 11083791 ER PT J AU Lei, BF Mackie, S Lukomski, S Musser, JM AF Lei, BF Mackie, S Lukomski, S Musser, JM TI Identification and immunogenicity of group A Streptococcus culture supernatant proteins SO INFECTION AND IMMUNITY LA English DT Article ID GROUP-A STREPTOCOCCI; COMPLEMENT RECEPTOR TYPE-3; GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE; BINDING-PROTEIN; ESCHERICHIA-COLI; SECRETED PROTEIN; PLASMIN-BINDING; CLONING; SURFACE; EXPRESSION AB Extracellular proteins made by group A Streptococcus (GAS) play critical roles in the pathogenesis of human infections caused by this bacterium. Although many extracellular GAS proteins have been identified and characterized, there has been no systematic analysis of culture supernatant proteins. Proteins present in the culture supernatant of strains of serotype M1 (MGAS 5005) and M3 (MGAS 315) mutants lacking production of the major extracellular cysteine protease were separated by two-dimensional gel electrophoresis and identified by amino-terminal amino acid sequencing and interrogation of available databases, including a serotype M1 genome sequence. In the aggregate, amino-terminal amino acid sequence data for 66 protein spots were generated, 53 unique sequences were obtained, and 44 distinct proteins were identified. Sixteen of the 44 proteins had apparent secretion signal sequences and 27 proteins did not. Eight of the 16 proteins with apparent secretion signal sequences have not been previously described for GAS. Antibodies against most of the apparently secreted proteins were present in sera from mice infected subcutaneously with MGAS 5005 or MGAS 315. Humans with documented GAS infections (pharyngitis, acute rheumatic fever, and severe invasive disease) also had serum antibodies reacting with many of the apparently secreted proteins, indicating that they were synthesized in the course of GAS-human interaction. The genes encoding four of the eight previously undescribed and apparently secreted culture supernatant proteins were cloned, and the proteins were overexpressed in Escherichia cell. Western blot analysis with these recombinant proteins and sera from GAS-infected mice and humans confirmed the immunogenicity of these proteins. Taken together, the data provide new information about the molecular aspects of GAS-host interactions. C1 NIAID, Lab Human Bacterial Pathogenesis, Rocky Mt Labs, NIH, Hamilton, MT 59840 USA. Baylor Coll Med, Dept Pathol, Houston, TX 77030 USA. RP Musser, JM (reprint author), NIAID, Lab Human Bacterial Pathogenesis, Rocky Mt Labs, NIH, 903 S 4th St, Hamilton, MT 59840 USA. NR 54 TC 118 Z9 120 U1 1 U2 3 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD DEC PY 2000 VL 68 IS 12 BP 6807 EP 6818 DI 10.1128/IAI.68.12.6807-6818.2000 PG 12 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 403AH UT WOS:000167020000041 PM 11083799 ER PT J AU Morrison, SG Su, H Caldwell, HD Morrison, RP AF Morrison, SG Su, H Caldwell, HD Morrison, RP TI Immunity to murine Chlamydia trachomatis genital tract reinfection involves B cells and CD4(+) T cells but not CD8(+) T cells SO INFECTION AND IMMUNITY LA English DT Article ID OUTER-MEMBRANE PROTEIN; NITRIC-OXIDE SYNTHASE; GENE KNOCKOUT MICE; PROTECTIVE MONOCLONAL-ANTIBODIES; MOUSE PNEUMONITIS BIOVAR; ANTI-BACTERIAL ACTIVITY; FEMALE GUINEA-PIGS; DEFICIENT MICE; MEDIATED CYTOLYSIS; INFECTION AB CD4(+) T-helper type 1 (Th1) responses are essential for the resolution of a primary Chlamydia trachomatis genital tract infection; however, elements of the immune response that function in resistance to reinfection are poorly understood. Defining the mechanisms of immune resistance to reinfection is important because the elements of protective adaptive immunity are distinguished by immunological memory and high-affinity antigen recognition, both of which are crucial to the development of efficacious vaccines. Using in vivo antibody depletion of CD4(+) and CD8(+) T cells prior to secondary intravaginal challenge, me identified lymphocyte populations that functioned in resistance to secondary chlamydial infection of the genital tract. Depletion of either CD4(+) or CD8(+) T cells in immune wild-type C57BL/6 mice had a limited effect on resistance to reinfection. However, depletion of CD4(+) T cells, but not CD8(+) T cells, in immune B-cell-deficient mice profoundly altered the course of secondary infection. CD4-depleted B-cell-deficient mice were unable to resolve a secondary infection, shed high levels of infectious chlamydiae, and did not resolve the infection until 3 to 4 weeks following the discontinuation of anti-CD4 treatment. These findings substantiated a predominant role for CD4(+) T cells in host resistance to chlamydial reinfection of the female genital tract and demonstrated that CD8(+) T cells are unnecessary for adaptive immune resistance. More importantly, however, this study establishes a previously unrecognized but very significant role for B cells in resistance to chlamydial reinfection and suggests that B cells and CD4(+) T cells may function synergistically in providing immunity in this model of chlamydial infection. Whether CD4(+) T cells and B cells function independently or dependently is unknown, but definition of those mechanisms is fundamental to understanding optimum protective immunity and to the development of highly efficacious immunotherapies against chlamydial urogenital infections. C1 Montana State Univ, Dept Microbiol, Bozeman, MT 59717 USA. NIAID, Rocky Mt Labs, Intracellular Parasites Lab, NIH, Hamilton, MT 59840 USA. RP Morrison, RP (reprint author), Montana State Univ, Dept Microbiol, Lewis Hall Room 109, Bozeman, MT 59717 USA. FU NIAID NIH HHS [R01 AI038991, AI 38991, R56 AI038991] NR 60 TC 139 Z9 150 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD DEC PY 2000 VL 68 IS 12 BP 6979 EP 6987 DI 10.1128/IAI.68.12.6979-6987.2000 PG 9 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 403AH UT WOS:000167020000064 PM 11083822 ER PT J AU Chen, X Yang, D Shen, W Dong, HF Wang, JM Oppenheim, JJ Howard, OMZ AF Chen, X Yang, D Shen, W Dong, HF Wang, JM Oppenheim, JJ Howard, OMZ TI Characterization of chenodeoxycholic acid as an endogenous antagonist of the G-coupled formyl peptide receptors SO INFLAMMATION RESEARCH LA English DT Article DE chenodeoxycholic acid; chemotaxis; bile acids; formyl peptide receptor; formyl peptide receptor like 1. calcium flux ID N-FORMYLPEPTIDE RECEPTOR; TUMOR-NECROSIS-FACTOR; CHRONIC LIVER-DISEASE; SERUM AMYLOID-A; BILE-ACIDS; OBSTRUCTIVE-JAUNDICE; BILIARY OBSTRUCTION; HUMAN MONOCYTES; NUCLEAR RECEPTOR; NEUTROPHIL CHEMOTAXIS AB Objective and design: To demonstrate the role of bile acids in immune modulation we examined the ability of select bile acids to inhibit leukocyte migration and chemoattractant receptor function. Materials: To elucidate this mechanism, we employed primary human monocytes, neutrophils and cell lines transfected to express either the high affinity fMLP receptor (FPR) or the low affinity fMLP receptor like 1 (FPRL1). Treatment: Cells were treated with chenodeoxycholic acid (CDCA) and related bile acids in a 0-400 micromolar range. Method: Cell viability, chemotaxis and calcium flux analysis were preformed. Results: We observed that pathophysiological levels (less than or equal to 150 micromolar) of CDCA competitively inhibited H-3-fMLP binding to human monocytes, FPR and FPRL1 transfected cells. Additionally, CDCA reduced both the chemotactic and calcium flux responses induced by fMLP or "W" peptide. Further, CDCA inhibited anti-FPR antibody binding to monocytes. Conclusions: CDCA selectively inhibited human leukocyte chemotaxis and calcium flux induced by fMLP, but not other chemoattractants, suggesting a mechanism for inhibition of inflammation and suppression of innate immune response. C1 NCI, Frederick Canc Res & Dev Ctr, Mol Immunoregulat Lab, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, Intraumural Res Support Program, Frederick, MD 21702 USA. RP Howard, OMZ (reprint author), NCI, Frederick Canc Res & Dev Ctr, Mol Immunoregulat Lab, Bld 560,Rm 31-19, Frederick, MD 21702 USA. RI Howard, O M Zack/B-6117-2012; Chen, Xin/I-6601-2015 OI Howard, O M Zack/0000-0002-0505-7052; Chen, Xin/0000-0002-2628-4027 FU NCCIH NIH HHS [Y2-AT-9002]; NCI NIH HHS [N01-CO-56000] NR 57 TC 33 Z9 35 U1 0 U2 6 PU BIRKHAUSER VERLAG AG PI BASEL PA VIADUKSTRASSE 40-44, PO BOX 133, CH-4010 BASEL, SWITZERLAND SN 1023-3830 J9 INFLAMM RES JI Inflamm. Res. PD DEC PY 2000 VL 49 IS 12 BP 744 EP 755 DI 10.1007/s000110050656 PG 12 WC Cell Biology; Immunology SC Cell Biology; Immunology GA 399LN UT WOS:000166814100015 PM 11211928 ER PT J AU Nanki, T Lipsky, PE AF Nanki, T Lipsky, PE TI Lack of correlation between chemokine receptor and T(h)1/T(h)2 cytokine expression by individual memory T cells SO INTERNATIONAL IMMUNOLOGY LA English DT Article DE chemokine receptor; cytokine; human; rheumatoid arthritis; T(h)1; T(h)2 ID MACROPHAGE-DERIVED CHEMOKINE; HUMAN INTERLEUKIN-8 RECEPTOR; PROTEIN-COUPLED RECEPTOR; MOLECULAR-CLONING; FUNCTIONAL EXPRESSION; EOTAXIN RECEPTOR; T-HELPER-2 CELLS; SINGLE-CELL; RHEUMATOID-ARTHRITIS; PEPTIDE RECEPTORS AB Chemokine and chemokine receptor interactions may have important roles in leukocyte migration to specific immune reaction sites. Recently, it has been reported that CXC chemokine receptor (CXCR) 3 and CC chemokine receptor (CCR) 5 were preferentially expressed on T(h)1 cells, and CCR3 and CCR4 were preferentially expressed on T(h)2 cells. To investigate chemokine receptor expression by T-h subsets in vivo, we analyzed cytokine (IL-2, IL-4 and IFN-gamma) and chemokine receptor (CXCR3, CXCR4, CCR3, CCR4 and CCR5) mRNA expression by individual peripheral CD4(+) memory T cells after short-term stimulation, employing a single-cell RT-PCR method. This ex vivo analysis shows that the frequencies of cells expressing chemokine receptor mRNA were not significantly different between T(h)1 and T(h)2 cells in normal peripheral blood. To assess a potential role of in vivo stimulation, we also analyzed unstimulated rheumatoid arthritis synovial CD4(+) memory T cells. CXCR3, CXCR4, CCR3 and CCR5 expression was detected by individual synovial T cells, but the frequencies of chemokine receptor mRNA were not clearly different between T(h)1 and non-T(h)1 cells defined by expression of IFN-gamma or lymphotoxin-a mRNA in all RA patients. These data suggest that chemokine receptor expression does not identify individual memory T cells producing Th-defining cytokines and therefore chemokine receptor expression cannot be a marker for T(h)1 or T(h)2 cells in vivo. C1 Univ Texas, SW Med Ctr, Dept Internal Med, Dallas, TX 75235 USA. Univ Texas, SW Med Ctr, Harold C Simmons Arthrit Res Ctr, Dallas, TX 75235 USA. RP Lipsky, PE (reprint author), NIAMSD, Bldg 10,Room 9N228,10 Ctr Dr MSC 1820, Bethesda, MD 20892 USA. FU NIAMS NIH HHS [AR39169] NR 64 TC 56 Z9 58 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0953-8178 J9 INT IMMUNOL JI Int. Immunol. PD DEC PY 2000 VL 12 IS 12 BP 1659 EP 1667 DI 10.1093/intimm/12.12.1659 PG 9 WC Immunology SC Immunology GA 384WF UT WOS:000165968700004 PM 11099305 ER PT J AU Semba, RD Kumwenda, N Taha, TE Mtimavalye, L Broadhead, R Miotti, PG Eisinger, W Hoover, D Chiphangwi, JD AF Semba, RD Kumwenda, N Taha, TE Mtimavalye, L Broadhead, R Miotti, PG Eisinger, W Hoover, D Chiphangwi, JD TI Plasma and breast milk vitamin A as indicators of vitamin A status in pregnant women SO INTERNATIONAL JOURNAL FOR VITAMIN AND NUTRITION RESEARCH LA English DT Article DE Africa; human immunodeficiency virus infection; milk; pregnancy; retinol; vitamin A ID IMMUNODEFICIENCY-VIRUS INFECTION; BETA-CAROTENE; A-DEFICIENCY; MORTALITY; RETINOL AB Breast milk vitamin A is not well characterized as an indicator of vitamin A status in women with infections. A controlled trial of vitamin A, 3 mg retinol equivalent/day, was conducted among 697 pregnant women with human immunodeficiency virus (HIV) infection in Malawi which allowed comparison of plasma versus breast milk vitamin A as indicators of vitamin A status. Retinol concentrations were measured in plasma at baseline (18-28 weeks) and 38 weeks gestation and breast milk at 6 weeks post-partum. Plasam alpha (1)-acid glycoprotein (ACP) and C-reactive protein (CRP) were measured at baseline. Plasma retinol (geometric mean, SD) at 38 weeks was 0.72(0.44, 1.18) and 0.61 (0.38, 0.98) mu mol/L (P < 0.0002) and breast milk retinol was 1.32 (0.71, 2.43) and 0.95 (0.49, 1.82) mu mol/L ( P < 0.0001) in vitamin A and placebo groups, respectively. Women with elevated acute phase protein (AGP > 1 gm/L and/or CRP > 5 mg/L) at baseline who received vitamin A had significantly higher plasma and breast milk vitamin A at follow-up compared with placebo. Elevated acute phase proteins did not distinguish women with low body stores of vitamin A. Breast milk retinol appears to be a better indicator of vitamin A status than plasma retinol in women with infections. C1 Johns Hopkins Univ, Sch Med, Dept Epidemiol, Baltimore, MD USA. Johns Hopkins Univ, Sch Hyg & Publ Hlth, Baltimore, MD USA. NIAID, Bethesda, MD 20892 USA. Rutgers State Univ, Dept Stat, Piscataway, NJ 08855 USA. Univ Malawi, Coll Med, Dept Obstet & Gynaecol, Blantyre, Malawi. Univ Malawi, Coll Med, Dept Paediat & Child Hlth, Blantyre, Malawi. Johns Hopkins Univ, Sch Med, Dept Ophthalmol, Baltimore, MD 21205 USA. RP Semba, RD (reprint author), 550 N Broadway,Suite 700, Baltimore, MD 21205 USA. FU NIAID NIH HHS [N01-AI-35173-117]; NICHD NIH HHS [HD30042, HD32247] NR 24 TC 17 Z9 18 U1 0 U2 3 PU VERLAG HANS HUBER PI BERN 9 PA LANGGASS-STRASSE 76, CH-3000 BERN 9, SWITZERLAND SN 0300-9831 J9 INT J VITAM NUTR RES JI Int. J. Vitam. Nutr. Res. PD DEC PY 2000 VL 70 IS 6 BP 271 EP 277 DI 10.1024/0300-9831.70.6.271 PG 7 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 397GC UT WOS:000166684100001 PM 11214351 ER PT J AU Patel, V Ensley, JF Gutkind, JS Yeudall, WA AF Patel, V Ensley, JF Gutkind, JS Yeudall, WA TI Induction of apoptosis in head-and-neck squamous carcinoma cells by gamma-irradiation and bleomycin is p53-independent SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article ID TUMOR-SUPPRESSOR P53; WILD-TYPE P53; DNA-DAMAGE; IN-VIVO; ANTICANCER DRUGS; GENE-EXPRESSION; BAX EXPRESSION; UP-REGULATION; X-RAYS; BCL-2 AB We have examined the ability of w gamma -irradiation and bleomycin to induce apoptosis in a model system consisting of cell lines derived from naturally occurring human head-and-neck squamous-cell carcinomas with contrasting p53 status and expression levels of pro- and anti-apoptotic molecules. Following exposure to gamma -irradiation (20 Gy) or bleomycin (3.5 muM) for 0 to 96 hr, cells expressing either transcriptionally inactive mutant p53 (HN6) or a truncated p53 molecule (HN 19) underwent apoptosis, as assessed by fluorescence-activated cell sorting and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling, in contrast to cells that express wild-type p53 (HN30), suggesting that apoptosis induced by these agents occurs by p53-independent mechanisms. Apoptosis in HN6 and HN19 cells was preceded by a G(2)/M cell-cycle block, as analyzed by DNA content and BrdU staining. In contrast, HN30 cells remained blocked in both G(1) and G(2)/M and failed to re enter the cell cycle. Levels of Bcl-2 were elevated in 3 of 10 cell lines, and only marginal differences were observed for Bcl-x(L). Pro-apoptotic proteins bax and Bcl-x(S) were detectable in normal keratinocytes and 4 tumor cell lines. Bax-delta (16 kDa) was highly represented in normal keratinocytes, and levels of bak were variable between cell lines. Elevated expression of Bcl-2 failed to protect HN19 cells from either gamma -irradiation or bleomycin-induced apoptosis. Our data support the existence of p53- and Bcl-2-independent pathways regulating apoptosis in keratinocytes and suggest that efficacy of either radiotherapy or bleomycin treatment for oral squamous-cell neoplasms may not, therefore, be influenced solely by endogenous p53 status. Published 2000 Wiley-Liss, Inc.(dagger). C1 NIDCR, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD 20892 USA. Wayne State Univ, Karmanos Canc Inst, Div Hematol Oncol, Detroit, MI USA. RP Patel, V (reprint author), NIDCR, Oral & Pharyngeal Canc Branch, NIH, 30 Convent Dr,MSC 4340,Bldg 30,Room 211, Bethesda, MD 20892 USA. RI Gutkind, J. Silvio/A-1053-2009 NR 43 TC 21 Z9 22 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD DEC 1 PY 2000 VL 88 IS 5 BP 737 EP 743 DI 10.1002/1097-0215(20001201)88:5<737::AID-IJC9>3.0.CO;2-7 PG 7 WC Oncology SC Oncology GA 370XW UT WOS:000165149400009 PM 11072242 ER PT J AU Fais, F Fronza, G Roncella, S Inga, A Campomenosi, P Cutrona, G Pezzolo, A Fedeli, F Abbondandolo, A Chiorazzi, N Pistoia, V Ferrarini, M AF Fais, F Fronza, G Roncella, S Inga, A Campomenosi, P Cutrona, G Pezzolo, A Fedeli, F Abbondandolo, A Chiorazzi, N Pistoia, V Ferrarini, M TI Analysis of stepwise genetic changes in an AIDS-related Burkitt's lymphoma SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article ID EPSTEIN-BARR-VIRUS; C-MYC ONCOGENE; HUMAN B-CELLS; FUNCTIONAL ASSAY; NUCLEAR ANTIGEN-2; EXPRESSION; MUTATIONS; REGION; ACTIVATION; BLOOD AB In this study, immunoglobulin variable (Ig V) region genes, c-myc re-arrangement and sequence and p53 status were analyzed in clones derived from a Burkitt's lymphoma cell line (LAM) in which it was previously demonstrated that Epstein-Barr virus (EBV) infection occurred late during lymphomagenesis. Such evidence was based on the finding that 2 groups of cellular clones, characterized by the same c-myc re-arrangement but different EBV-fused termini, were obtained from the LAM cell line. The Ig V gene sequences were identical for the 2 groups of clones with different EBV-fused termini. The Ig variable heavy (V-H) gene sequence displayed a substantial accumulation of point mutations (but no intraclonal diversification), whereas the productive Ig V lambda (V-lambda) gene sequence was virtually unmutated. Studies on the Ig V kappa (V-kappa) locus suggested a receptor revision event (with a switch from kappa to lambda chain production) prior to EBV infection. Likewise, it was determined that the mutations observed in both p53 alleles and in the re-arranged c-mye gene occurred before EBV infection. Based on these findings, we present a model for the various steps of lymphomagenesis. It is proposed that stimulation by an antigen or a superantigen initially favored the clonal expansion and accumulation of other cytogenetic changes, including those involved in receptor editing. These events occurred prior to or during the germinal center (GC) phase of B-cell maturation. Thereafter, possibly upon exit of the cells from the GC, EBV infection occurred, further promoting lymphomagenesis. (C) 2000 Wiley-Liss. Inc. C1 Natl Canc Inst, Div Clin Immunol, Genoa, Italy. Natl Canc Inst, Mutagenesis Lab, Genoa, Italy. Univ Genoa, Dept Oncol Biol & Genet, Genoa, Italy. St Andrea Hosp, Div Pathol, La Spezia, Italy. Ist Giannina Gaslini, Lab Oncol, I-16148 Genoa, Italy. N Shore Univ Hosp, Dept Med, Manhasset, NY USA. NYU, Sch Med, Manhasset, NY USA. RP Fais, F (reprint author), IST, Ist Nazl Ric Canc, Serv Immunol Clin, Lgo R Benzi 10, I-16132 Genoa, Italy. RI Campomenosi, Paola/C-9729-2011; Fais, Franco/K-1096-2016; OI Fais, Franco/0000-0002-6643-7083; Campomenosi, Paola/0000-0002-8853-1134; Ferrarini, Manlio/0000-0002-6154-2570; Cutrona, Giovanna/0000-0002-3335-1101 FU NCI NIH HHS [CA81554, R01 CA29088] NR 41 TC 6 Z9 6 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD DEC 1 PY 2000 VL 88 IS 5 BP 744 EP 750 DI 10.1002/1097-0215(20001201)88:5<744::AID-IJC10>3.0.CO;2-E PG 7 WC Oncology SC Oncology GA 370XW UT WOS:000165149400010 PM 11072243 ER PT J AU Holford, TR Zheng, TZ Mayne, ST Zahm, SH Tessari, JD Boyle, P AF Holford, TR Zheng, TZ Mayne, ST Zahm, SH Tessari, JD Boyle, P TI Joint effects of nine polychlorinated biphenyl (PCB) congeners on breast cancer risk SO INTERNATIONAL JOURNAL OF EPIDEMIOLOGY LA English DT Article DE breast cancer; environmental epidemiology; PCB; logistic regression; ridge regression ID ENVIRONMENTAL ORGANOCHLORINE EXPOSURE; ADIPOSE-TISSUE; POLYMORPHISMS; ABUNDANCE; SERUM; WOMEN AB Background Polychlorinated biphenyls (PCB) have been a major environmental health concern because of their wide distribution and persistence in the environment. Estimating joint effects of all congeners in a single analysis is complicated by correlation among exposure levels, and the resulting collinearity makes the results difficult to interpret. Methods Patients with breast-related surgery at Yale-New Haven Hospital were interviewed using a standardized questionnaire, and breast adipose tissue samples were analysed for nine PCB congeners (74, 118, 138, 153, 156, 170, 180, 183, 187). The study recruited 490 women (304 cases and 186 controls) between 1994 and 1997. Logistic ridge regression was used to analyse the instability caused by collinearity. Results Although total PCB did not appear to be associated with breast cancer risk, significant differences in effect were observed among the nine congeners. Logistic ridge regression demonstrated a protective effect on breast cancer risk for a potentially anti-oestrogenic and dioxin-like congener, 156, while two phenobarbital, CYP1A and CYP2B inducers had an adverse effect, 180 and 183. This analysis also suggested that a protective effect for another phenobarbital congener, 153, was largely explained by instability caused by collinearity. Conclusions These results indicate that studies of PCB congeners and health require an in-depth statistical analysis in order to better understand the complex issues related to their collinearity. C1 Yale Univ, Sch Med, Dept Epidemiol & Publ Hlth, New Haven, CT 06520 USA. Natl Canc Inst, Bethesda, MD USA. Colorado State Univ, Dept Environm Hlth, Ft Collins, CO USA. European Inst Oncol, Milan, Italy. RP Holford, TR (reprint author), Yale Univ, Sch Med, Dept Epidemiol & Publ Hlth, 60 Coll St,POB 208034, New Haven, CT 06520 USA. RI Boyle, Peter/A-4380-2014; Zahm, Shelia/B-5025-2015 OI Boyle, Peter/0000-0001-6251-0610; FU NCI NIH HHS [CA-62986] NR 27 TC 45 Z9 46 U1 0 U2 4 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0300-5771 J9 INT J EPIDEMIOL JI Int. J. Epidemiol. PD DEC PY 2000 VL 29 IS 6 BP 975 EP 982 DI 10.1093/ije/29.6.975 PG 8 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 391NA UT WOS:000166361000003 PM 11101537 ER PT J AU Kulldorff, M Sinha, R Chow, WH Rothman, N AF Kulldorff, M Sinha, R Chow, WH Rothman, N TI Comparing odds ratios for nested subsets of dietary components SO INTERNATIONAL JOURNAL OF EPIDEMIOLOGY LA English DT Article DE data interpretation; statistical; diet; epidemiological methods; nutrition assessment ID HEART-DISEASE; COLON CANCER; RISK; CONSUMPTION; ALCOHOL; WINE; BEER; MEAT; FAT AB Background In nutritional epidemiology, it is often of interest to disentangle the risk of disease associated with related foods or nutrients, where the food items are in a nested arrangement within a larger group. To compare odds ratios (OR) derived from a standard quantile-based analysis can be misleading since the amounts consumed may differ substantially for different dietary components. Methods The authors applied different logistic regression models on a case-control study concerning the risk of colorectal adenomas due to meat and its different subsets such as white meat, red meat and well-done red meat. Results By calculating OR for a fixed amount of intake, the authors suggest a method for partitioning the risk of one dietary item into that associated with increasingly detailed sub-components. A graph is presented for illustrating such partitions in terms of both addition and substitution effects. Conclusions Odds ratios based on upper versus lower quantiles or percentiles are useful as they compare the risk between the upper and lower ends of the consumption range. A complimentary set of OR are those based on fixed amounts of consumption. These allow for direct comparisons between nested subgroups of dietary components, in order to disentangle the risk linked to specific groups of foods or nutrients. C1 Univ Connecticut, Sch Med, Dept Community Med, Div Biostat, Farmington, CT 06030 USA. NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. RP Kulldorff, M (reprint author), Univ Connecticut, Sch Med, Dept Community Med, Div Biostat, Farmington, CT 06030 USA. RI Kulldorff, Martin/H-4282-2011; Sinha, Rashmi/G-7446-2015; OI Sinha, Rashmi/0000-0002-2466-7462; Kulldorff, Martin/0000-0002-5284-2993 NR 13 TC 14 Z9 14 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0300-5771 J9 INT J EPIDEMIOL JI Int. J. Epidemiol. PD DEC PY 2000 VL 29 IS 6 BP 1060 EP 1064 DI 10.1093/ije/29.6.1060 PG 5 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 391NA UT WOS:000166361000014 PM 11101548 ER PT J AU Murakami, T Yamamoto, N AF Murakami, T Yamamoto, N TI Roles of chemokines and chemokine receptors in HIV-1 infection SO INTERNATIONAL JOURNAL OF HEMATOLOGY LA English DT Article DE HIV-1; chemokine; chemokine receptor; AIDS; therapeutics ID HUMAN-IMMUNODEFICIENCY-VIRUS; GP120 ENVELOPE GLYCOPROTEIN; CELL-SURFACE ASSOCIATION; SMALL-MOLECULE INHIBITOR; DISEASE PROGRESSION; CD4-INDEPENDENT INFECTION; TROPIC HIV-1; GENETIC RESTRICTION; TYPE-1 INFECTION; CORECEPTOR CCR-5 AB Human immunodeficiency virus type 1 (HIV-1) uses a coreceptor together with CD 1 to enter CD4(+) target cells. The chemokine receptors CXCR4 and CCR5 have been found to be the major coreceptors for T-cell line-tropic and macrophagetropic HIV-1 strains, respectively, although many other chemokine and orphan receptors have also been identified as potential coreceptors for HIV-1. Genetic analyses has revealed the importance of chemokine and chemokine receptor genes in disease progression. The discovery of coreceptors provides a more defined scheme for virus entry in which the HIV-1 envelope glycoprotein sequentially binds CD4 and coreceptor, leading to a membrane fusion reaction between the viral envelope and the plasma membrane of the target cell. It also provides the basis for HIV-1 cell tropism. The identification of HN coreceptors provides new opportunities for the development of anti-HIV therapy. Many coreceptor-based therapeutic approaches have been developed, some of which are currently in clinical trials. Int J Hematol. 2000;72:412-417. (C)2000 The Japanese Society of Hematology. C1 Tokyo Med & Dent Univ, Dept Microbiol, Fac Med, Bunkyo Ku, Tokyo 1138519, Japan. Tokyo Med & Dent Univ, Dept Mol Virol, Fac Med, Bunkyo Ku, Tokyo 1138519, Japan. NIAID, Mol Microbiol Lab, NIH, Bethesda, MD 20892 USA. RP Yamamoto, N (reprint author), Tokyo Med & Dent Univ, Dept Microbiol, Fac Med, Bunkyo Ku, 1-5-45 Yushima, Tokyo 1138519, Japan. NR 76 TC 14 Z9 20 U1 0 U2 5 PU CARDEN JENNINGS PUBL CO LTD PI CHARLOTTESVILLE PA BLAKE CTR, STE 200, 1224 W MAIN ST, CHARLOTTESVILLE, VA 22903 USA SN 0925-5710 J9 INT J HEMATOL JI Int. J. Hematol. PD DEC PY 2000 VL 72 IS 4 BP 412 EP 417 PG 6 WC Hematology SC Hematology GA 391CC UT WOS:000166335700004 PM 11197206 ER PT J AU Nikjoo, H Laughton, CA Terrissol, M Panyutin, IG Goodhead, DT AF Nikjoo, H Laughton, CA Terrissol, M Panyutin, IG Goodhead, DT TI A method for radioprobing DNA structures using Auger electrons SO INTERNATIONAL JOURNAL OF RADIATION BIOLOGY LA English DT Article ID AQUEOUS-SOLUTION; TRIPLE-HELIX; MOLECULAR-DYNAMICS; STRAND BREAKS; DAMAGE; RADIATION; I-125; SIMULATION; I125; D(T)N.D(A)N.D(T)N AB Purpose: To present a new method for radioprobing a DNA triple helix structure by Auger electrons emitted in the decay of I-125 using theoretical/computational approaches. Materials and methods: A Monte Carlo track structure method was used to simulate the damage to a tripler resulting from Auger electrons emitted in the decay of an incorporated I-125 atom in plasmid DNA. Comparison of the theoretical frequency distributions of single-strand breaks induced on the Pu and Py strands with the experimental data and a knowledge of the distances from the strand breaks to the iodine provide information on the structures otherwise difficult to obtain with X-ray crystallography. Results: In comparing theoretical frequency distributions of single-strand breaks with the experimental data it is found that the results are very sensitive to the conformation of the tripler model used. It is found that the best fit to the experimental data results from using a hybrid tripler model, in which the base-step geometry is A-like, while the sugar puckers adopt the B-like C2'-endo conformation. Conclusions: The approach and technique presented here represent a valuable new addition to the methods available for DNA structure determination since they provide information on medium-range structure otherwise difficult to obtain in the absence of X-ray crystallography. It is concluded that currently accepted models for tripler structure are not optimal, and a modified structure is proposed that fits the radioprobing results better, while maintaining agreement with the fibre diffraction and NMR data. Although the method has proved to be very useful for scoring alternative trial solutions, further studies combining experimental data from multiple iodine positions with track structure modelling are required for directing structural optimization. C1 MRC, Radiat & Genome Stabil Unit, Harwell OX11 0RD, Berks, England. Univ Nottingham, Sch Pharmaceut Sci, Nottingham NG7 2RD, England. Univ Toulouse 3, CPAT, F-31062 Toulouse, France. NIH, Dept Nucl Med, Bethesda, MD 20892 USA. RP Nikjoo, H (reprint author), MRC, Radiat & Genome Stabil Unit, Harwell OX11 0RD, Berks, England. RI Laughton, Charles/E-5667-2010 OI Laughton, Charles/0000-0003-4090-3960 NR 36 TC 9 Z9 9 U1 0 U2 1 PU TAYLOR & FRANCIS LTD PI LONDON PA 11 NEW FETTER LANE, LONDON EC4P 4EE, ENGLAND SN 0955-3002 J9 INT J RADIAT BIOL JI Int. J. Radiat. Biol. PD DEC PY 2000 VL 76 IS 12 BP 1607 EP 1615 PG 9 WC Biology; Nuclear Science & Technology; Radiology, Nuclear Medicine & Medical Imaging SC Life Sciences & Biomedicine - Other Topics; Nuclear Science & Technology; Radiology, Nuclear Medicine & Medical Imaging GA 383WF UT WOS:000165906800004 PM 11133042 ER PT J AU Barry, CE Wilson, M Lee, R Schoolnik, GK AF Barry, CE Wilson, M Lee, R Schoolnik, GK TI DNA microarrays and combinatorial chemical libraries: tools for the drug discovery pipeline SO INTERNATIONAL JOURNAL OF TUBERCULOSIS AND LUNG DISEASE LA English DT Article; Proceedings Paper CT 5th Annual Meeting of the North American Region of the International-Union-Against-Tuberculosis-and-Lung-Disease CY FEB 24-26, 2000 CL VANCOUVER, CANADA SP Int Union Against Tuberculosis & Lung Dis, N Amer Reg ID GENE-EXPRESSION PATTERNS; MYCOBACTERIUM-TUBERCULOSIS; COMPARATIVE GENOMICS; HYBRIDIZATION; BCG; IDENTIFICATION; COMPLEX C1 NIAID, TB Res Sect, Host Def Lab, NIH, Rockville, MD 20852 USA. Affymax Res Inst, Santa Clara, CA USA. Div Infect Dis & Geog Med, Stanford, CA USA. RP Barry, CE (reprint author), NIAID, TB Res Sect, Host Def Lab, NIH, Twinbrook 2,Room 239,12441 Parklawn Dr, Rockville, MD 20852 USA. RI Barry, III, Clifton/H-3839-2012; Lee, Richard/J-4997-2013 OI Lee, Richard/0000-0002-2397-0443 FU Intramural NIH HHS [Z01 AI000693-15]; NIAID NIH HHS [AI144826, AI35969] NR 22 TC 9 Z9 10 U1 0 U2 0 PU INT UNION AGAINST TUBERCULOSIS LUNG DISEASE (I U A T L D) PI PARIS PA 68 BOULEVARD SAINT-MICHEL,, 75006 PARIS, FRANCE SN 1027-3719 J9 INT J TUBERC LUNG D JI Int. J. Tuberc. Lung Dis. PD DEC PY 2000 VL 4 IS 12 SU 2 BP S189 EP S193 PG 5 WC Infectious Diseases; Respiratory System SC Infectious Diseases; Respiratory System GA 382KP UT WOS:000165821700017 PM 11144552 ER PT J AU Ginsberg, A Fauci, AS AF Ginsberg, A Fauci, AS TI Eliminating tuberculosis SO ISSUES IN SCIENCE AND TECHNOLOGY LA English DT Editorial Material C1 NIAID, Bethesda, MD 20892 USA. RP Ginsberg, A (reprint author), NIAID, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0748-5492 J9 ISSUES SCI TECHNOL JI Issues Sci. Technol. PD WIN PY 2000 VL 17 IS 2 BP 7 EP 8 PG 2 WC Engineering, Multidisciplinary; Engineering, Industrial; Multidisciplinary Sciences; Social Issues SC Engineering; Science & Technology - Other Topics; Social Issues GA 492MH UT WOS:000172171800005 ER PT J AU Baldwin, W AF Baldwin, W TI Postdoctoral training SO ISSUES IN SCIENCE AND TECHNOLOGY LA English DT Editorial Material C1 NIH, Bethesda, MD 20892 USA. RP Baldwin, W (reprint author), NIH, Bldg 10, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0748-5492 J9 ISSUES SCI TECHNOL JI Issues Sci. Technol. PD WIN PY 2000 VL 17 IS 2 BP 14 EP + PG 2 WC Engineering, Multidisciplinary; Engineering, Industrial; Multidisciplinary Sciences; Social Issues SC Engineering; Science & Technology - Other Topics; Social Issues GA 492MH UT WOS:000172171800012 ER PT J AU Ishikawa, Y Miller, RW Machinami, R Sugano, H Goto, M AF Ishikawa, Y Miller, RW Machinami, R Sugano, H Goto, M TI Atypical osteosarcomas in Werner syndrome (adult progeria) SO JAPANESE JOURNAL OF CANCER RESEARCH LA English DT Article DE osteosarcoma; Werner syndrome; pathology; premature aging; cancer predisposed syndrome ID SYNDROME GENE; JAPANESE; CANCER AB Werner syndrome (WS), adult progeria, is more common in Japan than elsewhere. It predisposes to osteosarcoma (OS) and five other rare tumors. To determine if and how OS is atypical in this genetic disorder, we studied the characteristics of ten Japanese cases with respect to clinical features, pathology, and radiographs, and compared them with a hospital series of 36 skeletal OS with the same atypical age-range, 35-57 years. The anatomic sites were also atypical: seven ankle/foot, two radius and one patella compared with only one at the ankle in the hospital series. The osteoblastic cell-type was about equally frequent in both series, but, among others than the three major subtypes, there was only one in WS as compared with 14 (39%) in the hospital series. The types of mutations were sought in five WS cases with OS. One showed no mutation at any of the ten known loci for Japanese, two were of type 4/4 and two of type 6/6. The mutations 4 and 6 have been found in 66% of alleles of WS cases in Japan. The increased frequency and unusual age and site distributions of OS in WS may be due to increased susceptibility, related to later-life leg ulcers, and weight-bearing on spindly ankles weakened by severe loss of lower limb subcutaneous tissue. C1 Inst Canc, Dept Pathol, Toshima Ku, Tokyo 1708455, Japan. NCI, Clin Genet Branch, Bethesda, MD 20892 USA. Tokyo Metropolitan Otsuka Hosp, Div Rheumat Dis, Toshima Ku, Tokyo 1700005, Japan. RP Ishikawa, Y (reprint author), Inst Canc, Dept Pathol, Toshima Ku, 1-37-1 Kami Ikebukuro, Tokyo 1708455, Japan. NR 30 TC 15 Z9 17 U1 0 U2 0 PU BUSINESS CENTER ACADEMIC SOCIETIES JAPAN PI TOKYO PA 5-16-9 HONKOMAGOME, BUNKYO-KU, TOKYO, 113-8633, JAPAN SN 0910-5050 J9 JPN J CANCER RES JI Jpn. J. Cancer Res. PD DEC PY 2000 VL 91 IS 12 BP 1345 EP 1349 PG 5 WC Oncology SC Oncology GA 387HG UT WOS:000166116700018 PM 11123436 ER PT J AU Wells, KC Pelham, WE Kotkin, RA Hoza, B Abikoff, HB Abramowitz, A Arnold, LE Cantwell, DP Conners, CK Del Carmen, R Elliott, G Greenhill, LL Hechtman, L Hibbs, E Hinshaw, SP Jensen, PS March, JS Swanson, JM Schiller, E AF Wells, KC Pelham, WE Kotkin, RA Hoza, B Abikoff, HB Abramowitz, A Arnold, LE Cantwell, DP Conners, CK Del Carmen, R Elliott, G Greenhill, LL Hechtman, L Hibbs, E Hinshaw, SP Jensen, PS March, JS Swanson, JM Schiller, E TI Psychosocial treatment strategies in the MTA study: Rationale, methods, and critical issues in design and implementation SO JOURNAL OF ABNORMAL CHILD PSYCHOLOGY LA English DT Review DE attention deficit/hyperactivity disorder; psychosocial treatment; parent training; school intervention; summer treatment program ID DEFICIT HYPERACTIVITY DISORDER; SELF-CONTROL THERAPY; MOTHER-ADOLESCENT INTERACTIONS; PARENT-CHILD INTERACTIONS; ADHD CHILDREN; STIMULANT MEDICATION; BEHAVIOR-THERAPY; MATERNAL PSYCHOPATHOLOGY; CLASSROOM PERFORMANCE; MULTIMODAL TREATMENT AB The Collaborative Multimodal Treatment Study of Children with Attention Deficit Hyperactivity Disorder (ADHD), the MTA, is the first multisite, cooperative agreement treatment study of children, and the largest psychiatric/psychological treatment trial ever conducted by the National Institute of Mental Health. It examines the effectiveness of Medication vs. Psychosocial treatment vs. their combination for treatment of ADHD and compares these experimental arms to each other and to routine community care. In a parallel group design, 579 (male and female) ADHD children, aged 7-9 years, Il months, were randomly assigned to one of the four experimental arms, and then received 14 months of prescribed treatment (or community care) with periodic reassessments. After delineating the theoretical and empirical rationales for Psychosocial treatment of ADHD, we describe the MTA's Psychosocial Treatment strategy applied to all children in two of the four experimental arms (Psychosocial treatment alone; Combined treatment). Psychosocial treatment consisted of three major components: a Parent Training component, a two-part School Intervention component, and a child treatment component anchored in an intensive Summer Treatment Program. Components were selected based on evidence of treatment efficacy and because they address comprehensive symptom targets, settings, comorbidities, and functional domains. We delineate key conceptual and logistical issues faced by clinical researchers in design and implementation of Psychosocial research with examples of how these issues were addressed in the MTA study. C1 Duke Univ, Med Ctr, Durham, NC 27710 USA. SUNY Buffalo, Buffalo, NY 14260 USA. Univ Calif Irvine, Irvine, CA USA. Purdue Univ, Lafayette, IN USA. NYU, Med Ctr, New York, NY 10016 USA. Emory Univ, Sch Med, Atlanta, GA USA. Ohio State Univ, Columbus, OH 43210 USA. NIMH, Rockville, MD 20857 USA. Univ Calif San Francisco, San Francisco, CA 94143 USA. Columbia Univ, New York State Psychiat Inst, New York, NY USA. Montreal Childrens Hosp, Montreal, PQ H3H 1P3, Canada. Univ Calif Berkeley, Berkeley, CA 94720 USA. US Dept Educ, Washington, DC USA. RP Wells, KC (reprint author), Duke Univ, Med Ctr, POB 3320, Durham, NC 27710 USA. OI Jensen, Peter/0000-0003-2387-0650 FU NIMH NIH HHS [U01-MH50453, MH50467, U01-MH50461, UO1 MH50440, UO1 MH50447, UO1 MH50461] NR 111 TC 95 Z9 96 U1 9 U2 18 PU KLUWER ACADEMIC/PLENUM PUBL PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 0091-0627 J9 J ABNORM CHILD PSYCH JI J. Abnorm. Child Psychol. PD DEC PY 2000 VL 28 IS 6 BP 483 EP 505 DI 10.1023/A:1005174913412 PG 23 WC Psychology, Clinical; Psychology, Developmental SC Psychology GA 373PZ UT WOS:000165299000002 PM 11104313 ER PT J AU Epstein, JN Conners, CK Erhardt, D Arnold, LE Hechtman, L Hinshaw, SP Hoza, B Newcorn, JH Swanson, JM Vitiello, B AF Epstein, JN Conners, CK Erhardt, D Arnold, LE Hechtman, L Hinshaw, SP Hoza, B Newcorn, JH Swanson, JM Vitiello, B TI Familial aggregation of ADHD characteristics SO JOURNAL OF ABNORMAL CHILD PSYCHOLOGY LA English DT Article DE ADHD; aggregation; MTA; parents ID DEFICIT HYPERACTIVITY DISORDER; RISK-FACTORS; PSYCHIATRIC COMORBIDITY; MAJOR DEPRESSION; ATTENTION; CHILDREN; PATTERNS; CHILDHOOD; RELATIVES; ETIOLOGY AB Patterns of familial aggregation of ADHD symptoms in parents of ADHD and non-ADHD children were examined. Within the ADHD sample, symptom aggregation was examined as a Function of biological relationship, parent and child gender, and children's comorbid diagnoses. Participants consisted of parents of 579 children with ADHD, Combined Type participating in the multimodal treatment study of children with ADHD and parents of 288 normal control participants. Adult symptoms of ADHD were measured by both self-report and report of a significant other. Results indicated that the parents of children with ADHD had higher ratings of inattention/cognitive problems, hyperactivity/restlessness, impulsivity/emotional lability, and lower self-concept than parents of children without ADHD on both self-report and other-report ratings. Within the ADHD sample of children, other-report ratings of inattention/cognitive problems and impulsivity/emotional lability were higher For biological parents compared to nonbiological parents whereas self-ratings were not related to biological status. These findings support previous research documenting familial aggregation of ADHD and appear to strengthen the hypothesis that there is a genetic contribution to ADHD. C1 Duke Univ, Med Ctr, Durham, NC 27710 USA. Pepperdine Univ, Malibu, CA 90265 USA. Ohio State Univ, Columbus, OH 43210 USA. Montreal Childrens Hosp, Montreal, PQ H3H 1P3, Canada. Purdue Univ, W Lafayette, IN 47907 USA. Mt Sinai Med Ctr, New York, NY 10029 USA. Univ Calif Irvine, Irvine, CA USA. NIMH, Rockville, MD 20857 USA. RP Epstein, JN (reprint author), Duke Univ, Med Ctr, Box 3431, Durham, NC 27710 USA. OI Newcorn, Jeffrey /0000-0001-8993-9337 NR 39 TC 45 Z9 45 U1 4 U2 9 PU KLUWER ACADEMIC/PLENUM PUBL PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 0091-0627 J9 J ABNORM CHILD PSYCH JI J. Abnorm. Child Psychol. PD DEC PY 2000 VL 28 IS 6 BP 585 EP 594 DI 10.1023/A:1005187216138 PG 10 WC Psychology, Clinical; Psychology, Developmental SC Psychology GA 373PZ UT WOS:000165299000008 PM 11104319 ER PT J AU Rogers, AS Ellenberg, JH Douglas, SD Henry-Reid, L Peralta, L Wilson, CM AF Rogers, AS Ellenberg, JH Douglas, SD Henry-Reid, L Peralta, L Wilson, CM CA Adolescent Med HIV AIDS Res Networ TI The prevalence of anergy in human immunodeficiency virus-infected adolescents and the association of delayed-type hypersensitivity with subject characteristics SO JOURNAL OF ADOLESCENT HEALTH LA English DT Article DE adolescent; anergy; delayed-type hypersensitivity; HIV infection ID SKIN-TEST ANERGY; HIV-INFECTION; TUBERCULIN POSITIVITY; IMMUNE FUNCTION; SEX-DIFFERENCES; VIRAL LOAD; PROGRESSION; HEMOPHILIA; RISK; AIDS AB Purpose: To examine the prevalence of anergy in HIV-infected adolescents and factors associated with its occurrence. Methods: Anergy was defined as less than 2mm induration to each of three intradermally applied antigens (Candida albicans, tetanus toroid, and mumps) between 24 and 96 hours in a population of HIV-infected adolescents aged 12-18 at entry in a national multicenter study of HIV disease progression. CD4(+) T-cell counts and plasma HIV-1 RNA were measured in quality controlled laboratories. Factors associated with the probability of anergy were examined with contingency table comparisons, tree-structured classification, and logistic regression analyses. Results: Overall prevalence of anergy in this clinic-based population of 167 was 11% [7% in males and 12% in females (p = 0.57)]. The sole significant predictor of anergy was decreased CD4(+) T-cell count (p = 0.005). Conclusion: The prevalence of anergy is low in this HIV-infected population compared to older infected cohorts. The occurrence of differential rates of anergy in particular age and sex groupings that may be related to intrinsic immunologic differences requires further study. (C) Society for Adolescent Medicine, 2000. C1 NICHD, CRMC, PAMAB, Bethesda, MD 20892 USA. Westat, Rockville, MD USA. Childrens Hosp Philadelphia, Philadelphia, PA 19104 USA. Univ Penn, Dept Pediat, Philadelphia, PA 19104 USA. Cook Cty Hosp, Dept Adolescent Med, Chicago, IL 60612 USA. Univ Maryland, Dept Pediat, Baltimore, MD 21201 USA. Univ Alabama, Dept Geog Med, Birmingham, AL USA. RP Rogers, AS (reprint author), NICHD, CRMC, PAMAB, 6100 Execut Blvd,Room 4B11,MSC 7510, Bethesda, MD 20892 USA. NR 37 TC 4 Z9 4 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 1054-139X J9 J ADOLESCENT HEALTH JI J. Adolesc. Health PD DEC PY 2000 VL 27 IS 6 BP 384 EP 390 PG 7 WC Psychology, Developmental; Public, Environmental & Occupational Health; Pediatrics SC Psychology; Public, Environmental & Occupational Health; Pediatrics GA 375EB UT WOS:000165385600002 ER PT J AU Penninx, BWJH Deeg, DJH van Eijk, JTM Beekman, ATF Guralnik, JM AF Penninx, BWJH Deeg, DJH van Eijk, JTM Beekman, ATF Guralnik, JM TI Changes in depression and physical decline in older adults: a longitudinal perspective SO JOURNAL OF AFFECTIVE DISORDERS LA English DT Article DE depression; physical decline; older adults ID CARDIOVASCULAR EVENTS; SERVICE UTILIZATION; IMMUNE-SYSTEM; LATER LIFE; DISABILITY; COMMUNITY; SYMPTOMS; PERFORMANCE; POPULATION; HEALTH AB Background: The impact of chronicity and changes in depression on physical decline over time in older persons has not been elucidated. Methods: This prospective cohort study of 2121 community-dwelling persons aged 55-85 years uses two measurement occasions of depression (CES-D scale) over 3 years to distinguish persons with chronic, remitted, or emerging depression and persons who were never depressed. physical function is assessed by self-reported physical ability as well as by observed performance on a short battery of tests. Results: After adjustment for baseline physical function, health status and sociodemographic factors, chronic depression was associated with significantly greater decline in self-reported physical ability over 3 years when compared to never depressed persons (odds ratio (OR) = 2.83, 95% confidence interval (CI)= 1.86-4.30). In the oldest old, but not in the youngest old, chronic depression was also significantly predictive of greater decline in observed physical performance over 3 years (OR = 2.22, 95% CI = 1.43-3.79). Comparable effects were found for older persons with emerging depression. persons with remitted depression did not have greater decline in reported physical ability or observed performance than persons who were never depressed. Conclusions: Our findings among community-dwelling older persons show that chronicity of depression has: a large impact on physical decline over time. Since persons with remitted depression did not have greater physical decline than never depressed persons, these findings suggest that early recognition and treatment of depression in older persons could be protective for subsequent physical decline. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Free Univ Amsterdam, Inst Res Extramural Med, NL-1081 BT Amsterdam, Netherlands. Free Univ Amsterdam, Dept Psychiat, NL-1081 BT Amsterdam, Netherlands. NIA, Epidemiol Demog & Biometry Program, Bethesda, MD 20892 USA. RP Penninx, BWJH (reprint author), Free Univ Amsterdam, Inst Res Extramural Med, Van der Boechorststr 7, NL-1081 BT Amsterdam, Netherlands. NR 36 TC 102 Z9 109 U1 3 U2 4 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0165-0327 J9 J AFFECT DISORDERS JI J. Affect. Disord. PD DEC PY 2000 VL 61 IS 1-2 BP 1 EP 12 DI 10.1016/S0165-0327(00)00152-X PG 12 WC Clinical Neurology; Psychiatry SC Neurosciences & Neurology; Psychiatry GA 384PW UT WOS:000165954900001 PM 11099735 ER PT J AU Thayer, JF Lane, RD AF Thayer, JF Lane, RD TI A model of neurovisceral integration in emotion regulation and dysregulation SO JOURNAL OF AFFECTIVE DISORDERS LA English DT Article DE neurovisceral integration; emotion regulation and dysregulation ID GENERALIZED ANXIETY DISORDER; HEART-RATE-VARIABILITY; ANTERIOR CINGULATE CORTEX; FACIAL EXPRESSIONS; NEURAL ACTIVATION; NERVOUS-SYSTEM; ATTENTION; AWARENESS; RESPONSES; BEHAVIOR AB In the present paper we present the outlines of a model that integrates autonomic, attentional, and affective systems into a functional and structural network that may help to guide us in our understanding of emotion regulation and dysregulation. We will emphasize the relationship between attentional regulation and affective processes and propose a group of underlying physiological systems that serve to integrate these functions in the service of self-regulation and adaptability of the organism. We will attempt to place this network in the context of dynamical systems models which involve feedback and feedforward circuits with special attention to negative feedback mechanisms, inhibitory processes, and their role in response selection. From a systems perspective, inhibitory processes can be viewed as negative feedback circuits that allow for the interruption of ongoing behavior and the re-deployment of resources to other tasks. When these negative feedback mechanisms are compromised, positive feedback loops may develop as a result (of dis-inhibition), From this perspective, the relative sympathetic activation seen in anxiety disorders may represent dis-inhibition due to faulty inhibitory mechanisms. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIA, Gerontol Res Ctr, Baltimore, MD 21224 USA. Univ Arizona, Dept Psychiat, Tucson, AZ 85724 USA. RP Thayer, JF (reprint author), NIA, Gerontol Res Ctr, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. EM thayer@lpc.grc.nia.nih.gov RI Frank, David/E-8213-2012 NR 72 TC 695 Z9 707 U1 20 U2 101 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0165-0327 J9 J AFFECT DISORDERS JI J. Affect. Disord. PD DEC PY 2000 VL 61 IS 3 BP 201 EP 216 DI 10.1016/S0165-0327(00)00338-4 PG 16 WC Clinical Neurology; Psychiatry SC Neurosciences & Neurology; Psychiatry GA 398FD UT WOS:000166745500006 PM 11163422 ER PT J AU Roca-Ferrer, J Mullol, J Perez, M Xaubet, A Molins, L de Haro, J Shelhamer, J Picado, C AF Roca-Ferrer, J Mullol, J Perez, M Xaubet, A Molins, L de Haro, J Shelhamer, J Picado, C TI Effects of topical glucocorticoids on in vitro lactoferrin glandular secretion: Comparison between human upper and lower airways SO JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY LA English DT Article DE nasal mucosa; bronchial mucosa; lactoferrin; glandular secretion; budesonide; beclomethasone dipropionate; lipocortin 1; mucus hypersecretion; methacholine; glucocorticoids ID BECLOMETHASONE DIPROPIONATE; ALLERGIC RHINITIS; GLYCOCONJUGATE SECRETION; ANTIINFLAMMATORY DRUGS; EOSINOPHIL SURVIVAL; NASAL SECRETIONS; INVITRO; BUDESONIDE; ASTHMA; LIPOCORTIN-1 AB Background: Mucus hypersecretion is a hallmark of upper and lower airway diseases, such as rhinitis, asthma, and chronic obstructive pulmonary disease. Although topical glucocorticoids are widely used to treat mucosal inflammation, their effect on mucus hypersecretion remains uncertain. Objective: The aim of this study was to investigate the effect of budesonide and beclomethasone dipropionate on in vitro lactoferrin glandular secretion from both human nasal and bronchial mucosa and the potential mediating role of lipocortin 1. Methods: Nasal and bronchial explants obtained from patients undergoing surgery mere cultured in a controlled atmosphere. Lactoferrin (ELISA) was measured in culture supernatants, and lipocortin 1 (Western blot) was analyzed in explant tissues, Results: Both budesonide and beclomethasone dipropionate (10(-6) mol/L) decreased spontaneous lactoferrin secretion in nasal and bronchial mucosa, The maximum effect of corticosteroids (10(-6) mol/L) was obtained at day 3 in bronchial mucosa (budesonide: -56% +/- 9%, P < .05; beclomethasone dipropionate: 32% +/- 6%, P < .05) and at day 5 in nasal mucosa (budesonide: 34% +/- 10%, P < .05; beclomethasone dipropionate: -37% +/- 10%, P < .05). Methacholine (10(-4) mol/L) increased lactoferrin secretion in both bronchial (248% +/- 72%, P < .05) and nasal (107% +/- 28%, P < .05) explants, with this effect being completely abrogated by atropine, Budesonide caused a dose-related inhibitory effect on methacholine-induced lactoferrin secretion that was similar in both bronchial (down to -86% at 10(-6) mol/L) and nasal (down to -73% at 10(-6) mol/L) mucosa. Budesonide (10(-6) mol/L) did not show any effect on lipocortin 1 expression. Conclusions: These results suggest that glucocorticoid effects on airway inflammation may include a reduction of mucus hypersecretion in both nasal and bronchial mucosa. C1 Hosp Clin, Inst Clin Pneumol & Cirurgia Torac, Barcelona 08036, Spain. Inst Invest Biomed August Pi Sunyer, IDIBAPS, Barcelona, Spain. Hosp Clin, Serv Otorinolaringol, Barcelona 08036, Spain. Hosp Sargrat Cor, Serv Cirurgia Torac, Barcelona, Spain. Hosp Municipal Badalona, Serv Otorinolaringol, Barcelona, Spain. NIH, Ctr Clin, Dept Crit Care Med, Bethesda, MD 20892 USA. RP Picado, C (reprint author), Hosp Clin, Inst Clin Pneumol & Cirurgia Torac, Villarroel 170, Barcelona 08036, Spain. NR 46 TC 12 Z9 13 U1 0 U2 2 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0091-6749 J9 J ALLERGY CLIN IMMUN JI J. Allergy Clin. Immunol. PD DEC PY 2000 VL 106 IS 6 BP 1053 EP 1062 PG 10 WC Allergy; Immunology SC Allergy; Immunology GA 384ER UT WOS:000165930300005 PM 11112886 ER PT J AU Phipatanakul, W Eggleston, PA Wright, EC Wood, RA AF Phipatanakul, W Eggleston, PA Wright, EC Wood, RA CA Natl Cooperative Inner City Asthma TI Mouse allergen. II. The relationship of mouse allergen exposure to mouse sensitization and asthma morbidity in inner-city children with asthma SO JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY LA English DT Article DE mouse allergen; indoor allergens; inner-city asthma; sensitization; asthma morbidity ID DER-P-I; MITE; RISK; CHILDHOOD; CAT AB Background: Although mouse allergen is known to cause occupational asthma in laboratory workers, its potential significance in home environments has never been studied. Objective: This study was designed to define the prevalence of mouse sensitivity and its relationship to mouse allergen exposure and disease activity in inner-city children with asthma, Methods: A subset Of 499 subjects from the National Cooperative Inner-City Asthma Study had dust samples adequate for mouse allergen analysis, as well as valid puncture skin test (PST) results, Data were analyzed to relate mouse allergen exposure and other risk factors to mouse sensitization and asthma morbidity, Results: Eighty-nine (18%) of the 499 children had a positive mouse skin test response, Children whose homes had mouse allergen levels above the median (1.60 mug/g) in the kitchen had a significantly higher rate of mouse sensitization (23% vs 11%, P = .007). Atopy was also significantly related to mouse sensitization, with 40% of those with more than 4 positive PST responses having mouse sensitivity compared with 4% of those with no other positive PST responses (P < .0001). When atopy and exposure were considered together, 53% of those with more than 4 positive PST responses and allergen levels above the median had a positive PST response to mouse allergen compared with 22% of those with more than 4 positive PST responses and allergen levels below the median (P < .0001). The relationship among mouse allergen exposure, sensitization, and any measures of asthma morbidity was not statistically significant, Conclusions: Mouse allergen may be an important indoor allergen in inner-city children with asthma, with exposure and atopy contributing to mouse sensitization. C1 Harvard Univ, Sch Med, Childrens Hosp, Dept Pediat,Div Allergy & Immunol, Boston, MA USA. Johns Hopkins Univ, Sch Med, Dept Pediat, Div Allergy & Immunol, Baltimore, MD USA. New England Res Inst, Watertown, MA 02172 USA. Albert Einstein Sch Med, Bronx, NY USA. Childrens Mem Hosp, Chicago, IL 60614 USA. Cook Cty Hosp, Chicago, IL 60612 USA. Rainbow Babies & Childrens Hosp, Cleveland, OH 44106 USA. Henry Ford Hosp, Detroit, MI 48202 USA. Wayne State Univ, Med Ctr, Detroit, MI 48202 USA. Mt Sinai Sch Med, New York, NY USA. Washington Univ, Sch Med, St Louis, MO USA. St Louis Univ, Sch Med, St Louis, MO USA. Howard Univ, Washington, DC 20059 USA. NIAID, Program Off, Bethesda, MD 20892 USA. Ctr Occupat Environm Hlth, Irvine, CA USA. New England Res Inst, Watertown, MA 02172 USA. RP Wood, RA (reprint author), Johns Hopkins Hosp, CMSC 1102,600 N Wolfe St, Baltimore, MD 21287 USA. FU NIAID NIH HHS [AI07007]; NIEHS NIH HHS [ES09606] NR 23 TC 147 Z9 148 U1 0 U2 1 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0091-6749 J9 J ALLERGY CLIN IMMUN JI J. Allergy Clin. Immunol. PD DEC PY 2000 VL 106 IS 6 BP 1075 EP 1080 PG 6 WC Allergy; Immunology SC Allergy; Immunology GA 384ER UT WOS:000165930300008 PM 11112889 ER PT J AU Vanchieri, C AF Vanchieri, C TI National Cancer Institute (NCI) invites CAM practitioners to submit their data SO JOURNAL OF ALTERNATIVE AND COMPLEMENTARY MEDICINE LA English DT News Item C1 NCI, Bethesda, MD 20892 USA. RP Vanchieri, C (reprint author), NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1075-5535 J9 J ALTERN COMPLEM MED JI J. Altern. Complement Med. PD DEC PY 2000 VL 6 IS 6 BP 567 EP 568 PG 2 WC Integrative & Complementary Medicine SC Integrative & Complementary Medicine GA 387YR UT WOS:000166151100015 ER PT J AU Vanchieri, C AF Vanchieri, C TI National Center for Complementary and Alternative Medicine (NCCAM) announces intramural research program SO JOURNAL OF ALTERNATIVE AND COMPLEMENTARY MEDICINE LA English DT News Item C1 NCI, Bethesda, MD 20892 USA. RP Vanchieri, C (reprint author), NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1075-5535 J9 J ALTERN COMPLEM MED JI J. Altern. Complement Med. PD DEC PY 2000 VL 6 IS 6 BP 568 EP 569 PG 2 WC Integrative & Complementary Medicine SC Integrative & Complementary Medicine GA 387YR UT WOS:000166151100016 ER PT J AU Horikawa, I Yawata, T Barrett, JC AF Horikawa, I Yawata, T Barrett, JC TI Cellular senescence mechanisms independent of telomere shortening and telomerase: Other barriers to cell immortalization and carcinogenesis SO JOURNAL OF ANTI-AGING MEDICINE LA English DT Article ID HUMAN-DIPLOID FIBROBLASTS; HUMAN TUMOR-CELLS; MEDIATED CHROMOSOME TRANSFER; HUMAN CANCER-CELLS; OXIDATIVE STRESS; GROWTH ARREST; DNA-DAMAGE; REPLICATIVE SENESCENCE; PREMATURE SENESCENCE; CATALYTIC SUBUNIT AB Normal human somatic cells stop dividing after a limited number of cell divisions through the process termed "cellular senescence," which may function as a tumor-suppressive mechanism. Although the regulation of telomerase activity and telomere length plays an important role in cellular senescence and immortalization in human cells, recent findings from the telomerase introduction experiment, chromosome transfer experiment and studies on the inducers of premature senescence suggest that there are multiple mechanisms to induce cellular senescence, some of which are independent of telomerase and telomere regulation. Inactivation of the function of cell cycle regulatory p16(INK4A)/Rb pathway is a key event in immortalization. In rodents, which have long telomeres and telomerase activity, telomere shortening is not correlated with cellular senescence. These models may be useful to elucidate telomere-independent pathways in human cells. Studies in this field will clarify the barriers to human cell immortalization and carcinogenesis. C1 NCI, Lab Biosyst & Canc, NIH, Bethesda, MD 20892 USA. NCI, Canc & Aging Sect, NIH, Bethesda, MD 20892 USA. Natl Inst Environm Hlth Sci, Mol Carcinogenesis Lab, Res Triangle Pk, NC USA. RP Horikawa, I (reprint author), NCI, Lab Biosyst & Canc, NIH, 900 Rockvolle Pke,Bldg 40,Rm 2607, Bethesda, MD 20892 USA. NR 88 TC 6 Z9 6 U1 1 U2 1 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1094-5458 J9 J ANTI-AGING MED JI J. Anti-Aging Med. PD WIN PY 2000 VL 3 IS 4 BP 373 EP 382 DI 10.1089/rej.1.2000.3.373 PG 10 WC Geriatrics & Gerontology SC Geriatrics & Gerontology GA 397LP UT WOS:000166697400004 ER PT J AU Egan, SM Pease, AJ Lang, J Li, X Rao, V Gillette, WK Ruiz, R Ramos, JL Wolf, RE AF Egan, SM Pease, AJ Lang, J Li, X Rao, V Gillette, WK Ruiz, R Ramos, JL Wolf, RE TI Transcription activation by a variety of AraC/XylS family activators does not depend on the class II-specific activation determinant in the N-terminal domain of the RNA polymerase alpha subunit SO JOURNAL OF BACTERIOLOGY LA English DT Article ID SUPEROXIDE-INDUCIBLE GENES; ESCHERICHIA-COLI ADA; DNA-BINDING; SIGMA(70) SUBUNIT; ALKA PROMOTER; PROTEIN; MARA; IDENTIFICATION; OPERON; SOXS AB The N-terminal domain of the RNA polymerase or subunit (alpha -NTD) was tested for a role in transcription activation by a variety of AraC/XylS family members. Based on substitutions at residues 162 to 165 and an extensive genetic screen we conclude that alpha -NTD is not an activation target for these activators. C1 Univ Kansas, Dept Mol Biosci, Lawrence, KS 66045 USA. Univ Maryland Baltimore Cty, Dept Biol Sci, Baltimore, MD 21250 USA. NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. Dept Plant Biochem & Mol & Cellular Biol Plants, CSIC, Estac Expt Zaidin, Granada, Spain. RP Egan, SM (reprint author), Univ Kansas, Dept Mol Biosci, Lawrence, KS 66045 USA. OI Ramos, Juan L./0000-0002-8731-7435 FU NIGMS NIH HHS [GM55099, R01 GM027113, R29 GM055099, R01 GM055099, GM27113] NR 32 TC 13 Z9 14 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0021-9193 J9 J BACTERIOL JI J. Bacteriol. PD DEC PY 2000 VL 182 IS 24 BP 7075 EP 7077 DI 10.1128/JB.182.24.7075-7077.2000 PG 3 WC Microbiology SC Microbiology GA 407WM UT WOS:000167293800029 PM 11092872 ER PT J AU Lin, CY Huang, PH Liao, WL Cheng, HJ Huang, CF Kuo, JC Patton, WA Massenburg, D Moss, J Lee, FJS AF Lin, CY Huang, PH Liao, WL Cheng, HJ Huang, CF Kuo, JC Patton, WA Massenburg, D Moss, J Lee, FJS TI ARL4, an ARF-like protein that is developmentally regulated and localized to nuclei and nucleoli SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ADP-RIBOSYLATION FACTOR; SACCHAROMYCES-CEREVISIAE; CHOLERA-TOXIN; DIFFERENTIAL EXPRESSION; BINDING-PROTEINS; PHOSPHOLIPASE-D; CELLS; FAMILY; IDENTIFICATION; SEGMENTATION AB ADP-ribosylation factors (ARFs) are highly conserved similar to 20-kDa guanine nucleotide-binding proteins that participate in both exocytic and endocytic vesicular transport pathways via mechanisms that are only partially understood. Although several ARF-like proteins (ARLs) are known, their biological functions remain unclear. To characterize its molecular properties, we cloned mouse and human ARL4 (mARL4 and hARL4) cDNA. The appearance of mouse ARL4 mRNA during embryonic development coincided temporally with the sequential formation of somites and the establishment of brain compartmentation. Using ARL4-specific antibody for immunofluorescence microscopy, we observed that endogenous mARL4 in cultured Sertoli and neuroblastoma cells was mainly concentrated in nuclei. When expressed in COS7 cells, ARL4-T34N mutant, predicted to exist with GDP bound, was concentrated in nucleoli. Yeast two-hybrid screening and in vitro protein-interaction assays showed that hARL4 interacted with importin-alpha through its C-terminal NLS region and that the interaction was not nucleotide-dependent. Like ARL2 and -3, recombinant hARL4 did not enhance cholera toxin-catalyzed auto-ADP-ribosylation. Its binding of guanosine 5'-O-(thiotriphosphate) was modified by phospholipid and detergent, and the N terminus of hARL4, like that of ARF, was myristoylated. Our findings suggest that ARL4, with its distinctive nuclear/nucleolar localization and pattern of developmental expression, may play a unique role(s) in neurogenesis and somitogenesis during embryonic development and in the early stages of spermatogenesis in adults. C1 Natl Taiwan Univ, Coll Med, Inst Mol Med, Taipei, Taiwan. Natl Taiwan Univ, Coll Med, Inst Pathol, Taipei, Taiwan. NHLBI, Pulm Crit Care Med Branch, NIH, Bethesda, MD 20892 USA. RP Lee, FJS (reprint author), 7 Chung Shan S Rd, Taipei, Taiwan. OI LEE, FANG-JEN/0000-0002-2167-2426; HUANG, PEI-HSIN/0000-0002-2414-1487 NR 42 TC 32 Z9 38 U1 0 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 1 PY 2000 VL 275 IS 48 BP 37815 EP 37823 DI 10.1074/jbc.M002470200 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 379AL UT WOS:000165618700069 PM 10980193 ER PT J AU Yang, TX Park, JM Arend, L Huang, YN Topaloglu, R Pasumarthy, A Praetorius, H Spring, K Briggs, JP Schnermann, J AF Yang, TX Park, JM Arend, L Huang, YN Topaloglu, R Pasumarthy, A Praetorius, H Spring, K Briggs, JP Schnermann, J TI Low chloride stimulation of prostaglandin E-2 release and cyclooxygenase-2 expression in a mouse macula densa cell line SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID NITRIC-OXIDE SYNTHASE; RENIN SECRETION; RENOVASCULAR HYPERTENSION; RAT-KIDNEY; TRANSPORT; PHOSPHORYLATION; INHIBITION; ACTIVATION; PRESSURE AB Reducing luminal NaCl concentration in the macula densa region of the nephron stimulates renin secretion, and this response is blocked by a specific inhibitor of cyclooxygenase-2 (COX-2) (Traynor, T. R., Smart, A., Briggs, J. P., and Schnermann, J. (1999) Am. J. Physiol. Renal Physiol, 277, F706-710), To study whether low NaCl activates COX-2 activity or expression we clonally derived a macula densa cell line (MMDD1 cells) from SV-40 transgenic mice using fluorescence-activated cell sorting of renal tubular cells labeled with segment-specific fluorescent lectins, MMDD1 cells express COX-2, bNOS, NKCC2, and ROMK, but not Tamm-Horsfall protein, and showed rapid Rb-86(+) uptake that was inhibited by a reduction in NaCl concentration and by bumetanide or furosemide. Isosmotic exposure of MMDD1 cells to low NaCl (60 mM) caused a prompt and time-dependent stimulation of prostaglandin E-2 (PGE(2)) release that was prevented by the COX-2 specific inhibitor NS-398 (10 muM). Reducing NaCl to 60 and 6 mM for 16 h increased COX-2 expression in a chloride-dependent fashion. Low NaCl phosphorylated p38 kinase within 30 min and ERK1/2 kinases within 15 min without changing total MAP kinase levels. Low NaCl-stimulated PGE(2) release and COX-2 expression was inhibited by SE 203580 and PD 98059 (10 muM), inhibitors of p38 and ERK kinase pathways. We conclude that low chloride stimulates PGE(2) release and COX-2 expression in MMDD1 cells through activation of MAP kinases. C1 NIDDK, NIH, Bethesda, MD 20892 USA. NHLBI, NIH, Bethesda, MD 20892 USA. Univ Michigan, Dept Urol, Ann Arbor, MI 48109 USA. Univ Rochester, Dept Pathol & Lab Med, Rochester, NY 14642 USA. RP Schnermann, J (reprint author), NIDDK, NIH, Bldg 10,Rm 4D51,10 Ctr Dr,MSC 1370, Bethesda, MD 20892 USA. RI Briggs, Josephine/B-9394-2009 OI Briggs, Josephine/0000-0003-0798-1190 FU NIDDK NIH HHS [DK37448, DK39255, DK40042] NR 46 TC 103 Z9 107 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 1 PY 2000 VL 275 IS 48 BP 37922 EP 37929 DI 10.1074/jbc.M006218200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 379AL UT WOS:000165618700083 PM 10982805 ER PT J AU Shirakawa, H Herrera, JE Bustin, M Postnikov, Y AF Shirakawa, H Herrera, JE Bustin, M Postnikov, Y TI Targeting of high mobility group-14/-17 proteins in chromatin is independent of DNA sequence SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID GROUP CHROMOSOMAL-PROTEINS; NUCLEOSOME CORES; HMG-17; HISTONE-H1; NUCLEUS; BINDING; GENE AB Chromosomal proteins high mobility group (HMG)-14 and HMG-17 are nucleosomal-binding proteins that unfold the chromatin fiber and enhance transcription from chromatin templates. Their intracellular organization is dynamic and related to both cell cycle and transcription. Here we examine possible mechanisms for targeting HMG-14/-17 to specific regions in chromatin. Chromatin immunoprecipitation assays indicate that HMG-17 protein is not preferentially associated with chromatin regions containing transcriptionally active genes, or any type of specific DNA. We used a modification of the random amplified polymorphic DNA method to analyze DNA in various HMG-14/-17 nucleosome complexes. We found that although HMG-14 or HMG-17 proteins preferentially associate with core particles in which the DNA has a low frequency of CG dinucleotides, the genome does not contain consensus sequences that serve as specific targeting sites for the binding of either HMG-14 or HMG-17 proteins to nucleosomes. We used size exclusion and ion exchange chromatography to demonstrate that nuclei contain a large portion of HMG-17 associated with other proteins in a multiprotein complex. We suggest that these complexes regulate the dynamic organization of HMG-14/-17 in the nucleus and serve to target the proteins to specific sites in chromatin. C1 NCI, Prot Sect, Div Basic Sci, Nihon Univ, Bethesda, MD 20892 USA. RP NCI, Prot Sect, Div Basic Sci, Nihon Univ, Bldg 37,Rm 3D-20,9000 Rockville Pike, Bethesda, MD 20892 USA. EM yupo@helix.nih.gov RI Shirakawa, Hitoshi/D-1406-2009; Bustin, Michael/G-6155-2015 NR 24 TC 33 Z9 33 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 EI 1083-351X J9 J BIOL CHEM JI J. Biol. Chem. PD DEC 1 PY 2000 VL 275 IS 48 BP 37937 EP 37944 DI 10.1074/jbc.M000989200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 379AL UT WOS:000165618700085 PM 10973947 ER PT J AU Back, LH Banerjee, RK AF Back, LH Banerjee, RK TI Estimated flow resistance increase in a spiral human coronary artery segment SO JOURNAL OF BIOMECHANICAL ENGINEERING-TRANSACTIONS OF THE ASME LA English DT Article ID HELICAL PIPE; LAMINAR-FLOW; ATHEROSCLEROSIS; TORSION; MODEL AB Coronary flow estimates were made for a spiral coronary artery segment (identified from a post-mortem replica casting) by using a modified Dean number based on the approximate coil radius of curvature, as suggested earlier. The estimates were found to correlate experimental pressure drop data for helical coiled tubes. Over a physiological range of mean Reynolds numbers from 100 to 400 for blood flow through main coronary arteries, estimates of the flow resistance increase relative to a straight lumen segment ranged from about 20 to 80 percent, and were of similar magnitude to those found in a flow study in a sinuous coronary vessel segment with no spiral. C1 CALTECH, Jet Prop Lab, Pasadena, CA 91109 USA. NIH, Bioengn & Phys Sci Program, Bethesda, MD 20892 USA. RP Back, LH (reprint author), CALTECH, Jet Prop Lab, 4800 Oak Grove Dr, Pasadena, CA 91109 USA. NR 28 TC 10 Z9 10 U1 0 U2 2 PU ASME-AMER SOC MECHANICAL ENG PI NEW YORK PA THREE PARK AVE, NEW YORK, NY 10016-5990 USA SN 0148-0731 J9 J BIOMECH ENG-T ASME JI J. Biomech. Eng.-Trans. ASME PD DEC PY 2000 VL 122 IS 6 BP 675 EP 677 DI 10.1115/1.1319661 PG 3 WC Biophysics; Engineering, Biomedical SC Biophysics; Engineering GA 404QE UT WOS:000167111300017 PM 11192391 ER PT J AU Sass, HJ Musco, G Stahl, SJ Wingfield, PT Grzesiek, S AF Sass, HJ Musco, G Stahl, SJ Wingfield, PT Grzesiek, S TI Solution NMR of proteins within polyacrylamide gels: Diffusional properties and residual alignment by mechanical stress or embedding of oriented purple membranes SO JOURNAL OF BIOMOLECULAR NMR LA English DT Article DE dipolar couplings; HIV-1 Nef; ubiquitin ID DIPOLAR COUPLINGS; HIV-1 NEF; FIELD; MACROMOLECULES; ANISOTROPY; RELAXATION; DISTANCES; UBIQUITIN; PHASE AB The diffusive properties of biomacromolecules within the aqueous phase of polyacrylamide gels are described. High quality NMR spectra can be obtained under such conditions. As compared to water, a fivefold reduction in the translational diffusion constant, but only a 1.6-fold decrease (1.4-fold increase) in amide-N-15 T-2 (T-1) are observed for human ubiquitin within a 10% acrylamide gel. Weak alignment of the solute macromolecules can be achieved within such gels by vertical or radial compression or by the embedding of magnetically oriented purple membrane fragments. The methods are applied to derive residual dipolar couplings for human HIV-1 Nef and ubiquitin. C1 Univ Basel, Bioctr, Dept Biol Struct, CH-4056 Basel, Switzerland. NIAMSD, NIH, Bethesda, MD 20892 USA. RP Grzesiek, S (reprint author), Univ Basel, Bioctr, Dept Biol Struct, Klingelbergstr 70, CH-4056 Basel, Switzerland. RI musco, giovanna/I-7122-2012 OI musco, giovanna/0000-0002-0469-2994 NR 17 TC 246 Z9 250 U1 1 U2 20 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0925-2738 J9 J BIOMOL NMR JI J. Biomol. NMR PD DEC PY 2000 VL 18 IS 4 BP 303 EP 309 DI 10.1023/A:1026703605147 PG 7 WC Biochemistry & Molecular Biology; Spectroscopy SC Biochemistry & Molecular Biology; Spectroscopy GA 381XN UT WOS:000165789500002 PM 11200524 ER PT J AU Bhandari, R Mathew, R Vijayachandra, K Visweswariah, SS AF Bhandari, R Mathew, R Vijayachandra, K Visweswariah, SS TI Tyrosine phosphorylation of the human guanylyl cyclase C receptor SO JOURNAL OF BIOSCIENCES LA English DT Article DE EphB1; guanylyl cyclase C; tyrosine phosphorylation ID HEAT-STABLE ENTEROTOXIN; TRANSMEMBRANE CONDUCTANCE REGULATOR; NATRIURETIC-PEPTIDE RECEPTOR; DEPENDENT PROTEIN-KINASE; CDNA CLONING; EXPRESSION; LOCALIZATION; FAMILY; DEPHOSPHORYLATION; DESENSITIZATION AB Tyrosine phosphorylation events are key components of several cellular signal transduction pathways. This study describes a novel method for identification of substrates for tyrosine kinases. Go-expression of the tyrosine kinase EphB1 with the intracellular domain of guanylyl cyclase C (GCG) in Escherichia coli cells resulted in tyrosine phosphorylation of GCC, indicating that GCC is a potential substrate for tyrosine kinases. Indeed, GGC expressed in mammalian cells is tyrosine phosphorylated, suggesting that tyrosine phosphorylation may play a role in regulation of GCC signalling. This is the first demonstration of tyrosine phosphorylation of any member of the family of membrane-associated guanylyl cyclases. C1 Indian Inst Sci, Dept Mol Reprod Dev & Genet, Bangalore 560012, Karnataka, India. NCI, Cellular Carcinogenesis & Tumor Promot Lab, Bethesda, MD 20892 USA. RP Visweswariah, SS (reprint author), Indian Inst Sci, Dept Mol Reprod Dev & Genet, Bangalore 560012, Karnataka, India. OI Bhandari, Rashna/0000-0003-3101-0204 NR 31 TC 12 Z9 12 U1 0 U2 0 PU INDIAN ACADEMY SCIENCES PI BANGALORE PA P B 8005 C V RAMAN AVENUE, BANGALORE 560 080, INDIA SN 0250-5991 J9 J BIOSCIENCE JI J. Biosci. PD DEC PY 2000 VL 25 IS 4 BP 339 EP 346 DI 10.1007/BF02703787 PG 8 WC Biology SC Life Sciences & Biomedicine - Other Topics GA 383CQ UT WOS:000165865900005 PM 11120586 ER PT J AU Kimes, BW AF Kimes, BW TI The Cancer Education Grant Program of the National Cancer Institute SO JOURNAL OF CANCER EDUCATION LA English DT Editorial Material C1 NCI, Off Ctr Training & Resources, NIH, Bethesda, MD 20892 USA. RP Kimes, BW (reprint author), NCI, Off Ctr Training & Resources, NIH, Bethesda, MD 20892 USA. NR 0 TC 2 Z9 2 U1 0 U2 1 PU HANLEY & BELFUS INC PI PHILADELPHIA PA 210 S 13TH ST, PHILADELPHIA, PA 19107 USA SN 0885-8195 J9 J CANCER EDUC JI J. Cancer Educ. PD WIN PY 2000 VL 15 IS 4 BP 194 EP 195 PG 2 WC Oncology; Education, Scientific Disciplines; Public, Environmental & Occupational Health SC Oncology; Education & Educational Research; Public, Environmental & Occupational Health GA 459DY UT WOS:000170236800002 PM 11199233 ER PT J AU Malide, D Ramm, G Cushman, SW Slot, JW AF Malide, D Ramm, G Cushman, SW Slot, JW TI Immunoelectron microscopic evidence that GLUT4 translocation explains the stimulation of glucose transport in isolated rat white adipose cells SO JOURNAL OF CELL SCIENCE LA English DT Article DE GLUT4; adipose cell; insulin action; protein A-gold (immunogold) ID SKELETAL-MUSCLE; SUBCELLULAR TRAFFICKING; INSULIN; ISOFORM; DISTINCT; LOCALIZATION; COMPARTMENT; EXPRESSION; PHOTOLABEL; VESICLES AB We used an improved cryosectioning technique in combination with quantitative immunoelectron microscopy to study GLUT4 compartments in isolated rat white adipose cells, We provide clear evidence that in unstimulated cells most of the GLUT4 localizes intracellularly to tubulovesicular structures clustered near small stacks of Golgi and endosomes, or scattered throughout the cytoplasm, This localization is entirely consistent with that originally described in brown adipose tissue, strongly suggesting that the GLUT4 compartments in white and brown adipose cells are morphologically similar Furthermore, insulin induces parallel increases (with similar magnitudes) in glucose transport activity, approximately 16-fold, and cell-surface GLUT4, approximately 12-fold, Concomitantly, insulin decreases GLUT4 equally from all intracellular locations, in agreement with the concept that the entire cellular GLUT4 pool contributes to insulin-stimulated exocytosis, In the insulin-stimulated state, GLUT4 molecules are not randomly distributed on the plasma membrane, but neither are they enriched in caveolae, Importantly, the total number of GLUT4 C-terminal epitopes detected by the immuno-gold method is not significantly different between basal and insulin-stimulated cells, thus arguing directly against a reported insulin-induced unmasking effect. These results provide strong morphological evidence (1) that GLUT4 compartments are similar in all insulin-sensitive cells and (2) for the concept that GLUT4 translocation almost fully accounts for the increase in glucose transport in response to insulin. C1 NIDDK, Expt Diabet Metab & Nutr Sect, Diabet Branch, NIH, Bethesda, MD 20892 USA. Univ Utrecht, Sch Med, Dept Cell Biol, NL-3584 CX Utrecht, Netherlands. RP Cushman, SW (reprint author), NIDDK, Expt Diabet Metab & Nutr Sect, Diabet Branch, NIH, Bethesda, MD 20892 USA. NR 30 TC 68 Z9 70 U1 0 U2 0 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0021-9533 J9 J CELL SCI JI J. Cell Sci. PD DEC PY 2000 VL 113 IS 23 BP 4203 EP 4210 PG 8 WC Cell Biology SC Cell Biology GA 384YR UT WOS:000165975300008 PM 11069765 ER PT J AU Quirion, R Hyde, T AF Quirion, R Hyde, T TI Special issue on Human Brain (part 1) - Introduction SO JOURNAL OF CHEMICAL NEUROANATOMY LA English DT Editorial Material C1 McGill Univ, Dept Psychiat, Douglas Hosp, Res Ctr, Verdun, PQ H4H 1R3, Canada. NIMH, Clin Brain Disorders Branch, IRP, NIH, Bethesda, MD 20892 USA. RP Quirion, R (reprint author), McGill Univ, Dept Psychiat, Douglas Hosp, Res Ctr, 6875 LaSalle Blvd, Verdun, PQ H4H 1R3, Canada. NR 0 TC 0 Z9 0 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0891-0618 J9 J CHEM NEUROANAT JI J. Chem. Neuroanat. PD DEC PY 2000 VL 20 IS 3-4 BP 205 EP 205 DI 10.1016/S0891-0618(00)00098-3 PG 1 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 404TP UT WOS:000167117200001 ER PT J AU Phillips, JL Walther, MM Pezzullo, JC Rayford, W Choyke, PL Berman, AA Lineman, WM Doppman, JL Gill, JR AF Phillips, JL Walther, MM Pezzullo, JC Rayford, W Choyke, PL Berman, AA Lineman, WM Doppman, JL Gill, JR TI Predictive value of preoperative tests in discriminating bilateral adrenal hyperplasia from an aldosterone-producing adrenal adenoma SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID PRIMARY HYPER-ALDOSTERONISM; POSTURAL STIMULATION TEST; UPRIGHT POSTURE; HYPERALDOSTERONISM; MANAGEMENT; DIAGNOSIS; LOCALIZATION; INFUSION; CT AB In primary hyperaldosteronism, discriminating bilateral adrenal hyperplasia (BAH) from an aldosterone-producing adenoma (APA) is important because adrenalectomy, which is usually curative in APA, is seldom effective in BAH. We analyzed the results from our most recent 7-yr series to evaluate the predictive value of preoperative noninvasive tests compared with adrenal vein sampling (AVS). Forty-eight patients with hypertensive hyperaldosteronism underwent bed side testing, computed tomography (CT) imaging, and AVS. Those in whom the results of AVS indicated APA underwent adrenalectomy. Twelve (30%) and 14 (34%) of 41 patients with APA had paradoxical falls with ambulation in plasma aldosterone concentration (PAC) and 18-hydroxycorticosterone (18-OH-B), respectively. Twenty-nine (70%) and 26 (65%) APA patients had a rise in PAC and 18-OH-B, respectively, as did all 8 BAH patients. Significant identifiers of BAH were supine PAC values less than 15 ng/dL (P = 0.04), an increase greater than 60% (P = 0.02) in PAC with ambulation, and supine 18-OH-B values less than 60 ng/dL (P = 0.04). CT imaging alone was not predictive for BAH or APA. In our population, patients with a positive bedside test result (e.g. a fall in PAC and/or 18-OH-B) and a unilateral adrenal nodule on CT (10 of 41 patients) could have proceeded directly to adrenalectomy for ATA. However, a positive bedside test result with a negative CT or a negative bedside test result regardless of CT findings required AVS to confirm the diagnosis and site of disease. C1 NCI, Urol Oncol Branch, Bethesda, MD 20892 USA. NHLBI, Hypertens Endocrine Branch, Bethesda, MD 20892 USA. NIH, Walter Magnusson Clin Ctr, Dept Radiol, Bethesda, MD 20892 USA. Georgetown Univ, Dept Pharmacol, Washington, DC USA. RP Phillips, JL (reprint author), NCI, Urol Canc Inst, Bldg 10,Room 2B47, Bethesda, MD 20892 USA. NR 27 TC 96 Z9 102 U1 0 U2 1 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0021-972X J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD DEC PY 2000 VL 85 IS 12 BP 4526 EP 4533 DI 10.1210/jc.85.12.4526 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 390CM UT WOS:000166277800017 PM 11134103 ER PT J AU Stratakis, CA Schussheim, DH Freedman, SM Keil, MF Pack, SD Agarwal, SK Skarulis, MC Weil, RJ Lubensky, IA Zhuang, ZP Oldfield, EH Marx, SJ AF Stratakis, CA Schussheim, DH Freedman, SM Keil, MF Pack, SD Agarwal, SK Skarulis, MC Weil, RJ Lubensky, IA Zhuang, ZP Oldfield, EH Marx, SJ TI Pituitary macroadenoma in a 5-year-old: An early expression of multiple endocrine neoplasia type 1 SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article; Proceedings Paper CT 82nd Annual Meeting of the Endocrine-Society CY JUN 21-24, 2000 CL TORONTO, CANADA SP Endocrine Soc ID TUMOR-SUPPRESSOR GENE; MEN1 GENE; ADENOMAS; MUTATIONS; CHROMOSOME-11; DELETIONS; DISEASE; CYCLASE; GS AB Multiple endocrine neoplasia type 1 (MEN 1) is associated with parathyroid, enteropancreatic, pituitary, and other tumors. The MEN1 gene, a tumor suppressor, is located on chromosome 11. Affected individuals inherit a mutated MEN1 allele, and tumorigenesis in specific tissues follows inactivation of the remaining MEN1 allele. MEN 1-associated endocrine tumors usually become clinically evident in late adolescence or young adulthood, as high levels of PTH, gastrin, or PRL. Because each of these tumors can usually be controlled with medications and/or surgery, MEN 1 has been regarded mainly as a treatable endocrinopathy of adults. Unlike in MEN 2, early testing of children in MEN 1 families is not recommended. We report a 2.3-cm pituitary macroadenoma in a 5-yr-old boy with familial MEN 1. He presented with growth acceleration, acromegaloid features, and hyperprolactinemia. We tested systematically to see whether his pituitary tumor had causes similar to or different from a typical MEN 1 turner. Germ line DNA of the propositus and his affected relatives revealed a heterozygous point mutation in the MEN1 gene, which leads to a His139Asp (H139D) amino acid substitution. The patient had no other detectable germ-line mutations on either MEN1 allele. DNA sequencing and fluorescent in situ hybridization with a MEN1 genomic DNA sequence probe each demonstrated one copy of the MEN1 gene to be deleted in the pituitary tumor and not in normal DNA, proving MEN1 "second hit" as a tumor cause. Gs alpha mutation, common in nonhereditary GH-producing tumors, was not detected in this tumor. We conclude that this pituitary macroadenoma showed molecular genetic features of a typical MEN 1-associated tumor. This patient represents the earliest presentation of any morbid endocrine tumor in MEN 1. A better understanding of early onset MEN 1 disease is needed to formulate recommendations for early MEN 1 genetic testing. C1 NICHHD, Dev Endocrinol Branch, Unit Genet & Endocrinol, NIH, Bethesda, MD 20892 USA. NIDDKD, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. NINDS, Surg Neurol Branch, NIH, Bethesda, MD 20892 USA. Joe DiMaggio Childrens Hosp, Pediat Specialty Ctr, Hollywood, FL 33021 USA. RP Stratakis, CA (reprint author), NICHHD, Dev Endocrinol Branch, Unit Genet & Endocrinol, NIH, Bldg 10,Room 10N262,MSC1862,10 Ctr Dr, Bethesda, MD 20892 USA. EM stratakc@cc1.nichd.nih.gov RI Pack, Svetlana/C-2020-2014; Agarwal, Sunita/D-1428-2016 OI Agarwal, Sunita/0000-0002-7557-3191 NR 43 TC 68 Z9 74 U1 0 U2 4 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0021-972X J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD DEC PY 2000 VL 85 IS 12 BP 4776 EP 4780 DI 10.1210/jc.85.12.4776 PG 5 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 390CM UT WOS:000166277800056 PM 11134142 ER PT J AU Stratakis, CA Kirschner, LS AF Stratakis, CA Kirschner, LS TI Isolated familial somatotropinomas: Does the disease map to 11q13 or to 2p16? SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Letter ID HETEROZYGOSITY; MUTATIONS; TUMORS; GENE C1 NICHHD, Unit Genet & Endocrinol, Dev Endocrinol Branch, NIH, Bethesda, MD 20892 USA. RP Stratakis, CA (reprint author), NICHHD, Unit Genet & Endocrinol, Dev Endocrinol Branch, NIH, Bldg 10,Room 10N262,10 Ctr Dr,MSC1862, Bethesda, MD 20892 USA. NR 7 TC 5 Z9 5 U1 0 U2 0 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0021-972X J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD DEC PY 2000 VL 85 IS 12 BP 4920 EP 4920 DI 10.1210/jc.85.12.4920 PG 1 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 390CM UT WOS:000166277800079 PM 11134164 ER PT J AU Klabunde, CN Potosky, AL Legler, JM Warren, JL AF Klabunde, CN Potosky, AL Legler, JM Warren, JL TI Development of a comorbidity index using physician claims data SO JOURNAL OF CLINICAL EPIDEMIOLOGY LA English DT Article DE comorbidity; claims data; administrative data; Medicare; breast cancer; prostate cancer ID ICD-9-CM ADMINISTRATIVE DATA; BREAST-CANCER; PROSTATE-CANCER; MEDICAL RECORD; CO-MORBIDITY; CARE; POPULATION; ACCURACY; SURVIVAL; DEATH AB Important comorbidities recorded on outpatient claims in administrative datasets may be missed in analyses when only inpatient care is considered. Using the comorbid conditions identified by Charlson and colleagues, we developed a comorbidity index that incorporates the diagnostic and procedure data contained in Medicare physician (Part B) claims. In the national cohorts of elderly prostate (n = 28,868) and breast cancer (n = 14,943) patients assessed in this study, less than 10% of patients had comorbid conditions identified when only Medicare hospital (Part A) claims were examined. By incorporating physician claims, the proportion of patients with comorbid conditions increased to 25%. The new physician claims comorbidity index significantly contributes to models of 2-year noncancer mortality and treatment received in both patient cohorts. We demonstrate the utility of a disease-specific index using an alternative method of construction employing study-specific weights. The physician claims index can be used in conjunction with a comorbidity index derived from inpatient hospital claims, or employed as a stand-alone measure. (C) 2000 Elsevier Science Inc. All rights reserved. C1 NCI, Hlth Serv & Econ Branch, Appl Res Program, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. NCI, Stat Res & Applicat Branch, Surveillance Res Program, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. RP Klabunde, CN (reprint author), NCI, Hlth Serv & Econ Branch, Appl Res Program, Div Canc Control & Populat Sci, Execut Plaza Room 4005,6130 Execut Blvd MSC 7344, Bethesda, MD 20892 USA. NR 42 TC 890 Z9 894 U1 5 U2 17 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0895-4356 J9 J CLIN EPIDEMIOL JI J. Clin. Epidemiol. PD DEC PY 2000 VL 53 IS 12 BP 1258 EP 1267 DI 10.1016/S0895-4356(00)00256-0 PG 10 WC Health Care Sciences & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA 385NG UT WOS:000166010100011 PM 11146273 ER PT J AU Forster, HP AF Forster, HP TI Legal trends in bioethics SO JOURNAL OF CLINICAL ETHICS LA English DT Article C1 NIH, Dept Clin Bioeth, Bethesda, MD 20892 USA. RP Forster, HP (reprint author), NIH, Dept Clin Bioeth, Bldg 10, Bethesda, MD 20892 USA. NR 31 TC 1 Z9 1 U1 0 U2 0 PU UNIV PUBLISHING GROUP PI HAGERSTOWN PA 17100 COLE RD #312, HAGERSTOWN, MD 21740-6901 USA SN 1046-7890 J9 J CLIN ETHIC JI J. Clin. Ethics PD WIN PY 2000 VL 11 IS 4 BP 373 EP 382 PG 10 WC Ethics; Social Sciences, Biomedical SC Social Sciences - Other Topics; Biomedical Social Sciences GA 408BW UT WOS:000167307500012 PM 11252921 ER PT J AU Yun, J Schoneberg, T Liu, J Schulz, A Ecelbarger, CA Promeneur, D Nielsen, S Sheng, H Grinberg, A Deng, CX Wess, J AF Yun, J Schoneberg, T Liu, J Schulz, A Ecelbarger, CA Promeneur, D Nielsen, S Sheng, H Grinberg, A Deng, CX Wess, J TI Generation and phenotype of mice harboring a nonsense mutation in the V2 vasopressin receptor gene SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article ID NEPHROGENIC DIABETES-INSIPIDUS; HUMAN ANTIDIURETIC-HORMONE; WATER CHANNEL; RAT-KIDNEY; AQUAPORIN-2; PATHOPHYSIOLOGY; EXPRESSION; CLONING; HYDRONEPHROSIS; MANIPULATION AB The V2 vasopressin receptor (V2R) plays a key role in the maintenance of a normal body water balance. To generate an in vivo model that allows the physiological and molecular analysis of the role of V2Rs in kidney function, we have created mouse lines that lack functional V2Rs by using targeted mutagenesis in mouse embryonic stem cells. Specifically, we introduced a nonsense mutation known to cause X-linked nephrogenic diabetes insipidus (XNDI) in humans (Glu242stop) into the mouse genome. V2R-deficient hemizygous male pups showed a decrease in basal urine osmolalities and were unable to concentrate their urine. These pups also exhibited an enlargement of renal pelvic space, failed to thrive, and died within the first week after birth due to hypernatremic dehydration. Interestingly, female mice heterozygous for the V2R mutation showed normal growth but displayed an XNDI-like phenotype, characterized by reduced urine concentrating ability of the kidney, polyuria, and polydipsia. Western blot analysis and immunoelectron microscopic studies showed that the loss of functional V2Rs had no significant effect on the basal expression levels of aquaporin-2 and the bumetanide-sensitive Na-K-2Cl cotransporter (BSC-1). The V2R mutant mice described here should serve as highly useful tools for the development of novel therapeutic strategies for the treatment of XNDI. C1 NIDDKD, Bioorgan Chem Lab, Bethesda, MD 20892 USA. Free Univ Berlin, Inst Pharmakol, Fachbereich Humanmed, D-1000 Berlin, Germany. Georgetown Univ, Dept Med, Div Endocrinol & Metab, Washington, DC 20057 USA. Aarhus Univ, Inst Anat, Dept Cell Biol, DK-8000 Aarhus, Denmark. NICHHD, Lab Mammalian Genes & Dev, Bethesda, MD 20892 USA. NIDDKD, Biochem & Metab Lab, Bethesda, MD 20892 USA. RP Wess, J (reprint author), NIDDKD, Bioorgan Chem Lab, Bldg 8A,Room B1A-05,8 Ctr Dr MSC 0810, Bethesda, MD 20892 USA. RI chen, xuanlan/H-4158-2011; deng, chuxia/N-6713-2016 NR 57 TC 63 Z9 63 U1 0 U2 2 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA ROOM 4570 KRESGE I, 200 ZINA PITCHER PLACE, ANN ARBOR, MI 48109-0560 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD DEC PY 2000 VL 106 IS 11 BP 1361 EP 1371 DI 10.1172/JCI9154 PG 11 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 379MZ UT WOS:000165647800008 PM 11104789 ER PT J AU Battey, JF AF Battey, JF TI A genetic approach to understanding inner ear function SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Editorial Material ID UNCONVENTIONAL MYOSIN; DEAFNESS; MUTATION; MOUSE; MICE C1 Natl Inst Deafness & Other Commun Disorders, Bethesda, MD 20892 USA. RP Battey, JF (reprint author), Natl Inst Deafness & Other Commun Disorders, NIH Bldg 31,Room 3C02,31 Convent Dr MSC 2320,9000, Bethesda, MD 20892 USA. NR 17 TC 1 Z9 2 U1 1 U2 3 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA ROOM 4570 KRESGE I, 200 ZINA PITCHER PLACE, ANN ARBOR, MI 48109-0560 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD DEC PY 2000 VL 106 IS 12 BP 1431 EP 1432 DI 10.1172/JCI11763 PG 2 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 383WW UT WOS:000165908500001 PM 11120747 ER PT J AU Lee, MP Ravenel, JD Hu, RJ Lustig, LR Tomaselli, G Berger, RD Brandenburg, SA Litzi, TJ Bunton, TE Limb, C Francis, H Gorelikow, M Gu, H Washington, K Argani, P Goldenring, JR Coffey, RJ Feinberg, AP AF Lee, MP Ravenel, JD Hu, RJ Lustig, LR Tomaselli, G Berger, RD Brandenburg, SA Litzi, TJ Bunton, TE Limb, C Francis, H Gorelikow, M Gu, H Washington, K Argani, P Goldenring, JR Coffey, RJ Feinberg, AP TI Targeted disruption of the Kvlqt1 gene causes deafness and gastric hyperplasia in mice SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article ID BECKWITH-WIEDEMANN-SYNDROME; POTASSIUM CHANNEL GENE; DEVELOPMENTAL-CHANGES; IMPRINTED GENE; PARIETAL-CELLS; MOUSE; MUTATION; HEART; VARIABILITY; EXPRESSION AB The K nu LQT1 gene encodes a voltage-gated potassium channel. Mutations in K nu LQT1 underlie the dominantly transmitted Ward-Romano long QT syndrome, which causes cardiac arrhythmia, and the recessively transmitted Jervell and Lange-Nielsen syndrome, which causes both cardiac arrhythmia and congenital deafness. K nu LQT1 is also disrupted by balanced germline chromosomal rearrangements in patients with Beckwith-Wiedemann syndrome (BWS), which causes prenatal overgrowth and cancer. Because of the diverse human disorders and organ systems affected by this gene, we developed an animal model by inactivating the murine K nu lqt1. No electrocardiographic abnormalities were observed. However, homozygous mice exhibited complete deafness, as well as circular movement and repetitive falling, suggesting imbalance. Histochemical study revealed severe anatomic disruption of the cochlear and vestibular end organs, suggesting that K nu lqt1 is essential for normal development of the inner ear. Surprisingly, homozygous mice also displayed threefold enlargement by weight of the stomach resulting from mucous neck cell hyperplasia. Finally, there were no features of BWS, suggesting that K nu lqt1 is not responsible for BWS. C1 Johns Hopkins Univ, Sch Med, Dept Med, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Inst Geriat Med, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Dept Mol Biol & Genet, Baltimore, MD 21205 USA. NIAID, Immunol Lab, Rockville, MD USA. Johns Hopkins Univ, Sch Med, Dept Otorhinolaryngol, Baltimore, MD USA. Johns Hopkins Univ, Sch Med, Dept Comparat Med, Baltimore, MD USA. Vanderbilt Univ, Med Ctr, Dept Pathol, Nashville, TN USA. Johns Hopkins Univ, Sch Med, Dept Pathol, Baltimore, MD USA. Med Coll Georgia, Inst Mol Med & Genet, Augusta, GA 30912 USA. Vet Affairs Med Ctr, Augusta, GA USA. RP Feinberg, AP (reprint author), Johns Hopkins Univ, Sch Med, Dept Med, 720 Rutland Ave,Ross 1064, Baltimore, MD 21205 USA. NR 37 TC 182 Z9 191 U1 0 U2 3 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA ROOM 4570 KRESGE I, 200 ZINA PITCHER PLACE, ANN ARBOR, MI 48109-0560 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD DEC PY 2000 VL 106 IS 12 BP 1447 EP 1455 DI 10.1172/JCI10897 PG 9 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 383WW UT WOS:000165908500006 PM 11120752 ER PT J AU Al-Mohsen, IZ Sutton, DA Sigler, L Almodovar, E Mahgoub, N Frayha, H Al-Hajjar, S Rinaldi, MG Walsh, TJ AF Al-Mohsen, IZ Sutton, DA Sigler, L Almodovar, E Mahgoub, N Frayha, H Al-Hajjar, S Rinaldi, MG Walsh, TJ TI Acrophialophora fusispora brain abscess in a child with acute lymphoblastic leukemia: Review of cases and taxonomy SO JOURNAL OF CLINICAL MICROBIOLOGY LA English DT Review ID FUNGAL-INFECTIONS; RISK-FACTORS; PHEOHYPHOMYCOSIS; ITRACONAZOLE; CANCER AB A 12-year-old girl with acute lymphoblastic leukemia was referred to King Faisal Specialist Hospital and Research Center. The diagnosis without central nervous system (CNS) involvement was confirmed on admission, and chemotherapy was initiated according to the Children Cancer Group (CCG) 1882 protocol for high-risk-group leukemia. During neutropenia amphotericin B (AMB) (1 mg/kg of body weight/day) was initiated for presumed fungal infection when a computed tomography (CT) scan of the chest revealed multiple nodular densities. After 3 weeks of AMB therapy, a follow-up chest CT revealed progression of the pulmonary nodules. The patient subsequently suffered a seizure, and a CT scan of the brain was consistent with infarction or hemorrhage. Because of progression of pulmonary lesions while receiving AMB, antifungal therapy was changed to liposomal AMB (LAMB) (6 mg/kg/day). Despite 26 days of LAMB, the patient continued to have intermittent fever, and CT and magnetic resonance imaging of the brain demonstrated findings consistent with a brain abscess. Aspiration of brain abscess was performed and the Gomori methenamine silver stain was positive for hyphal elements. Culture of this material grew Acrophialophora fusispora. Lung biopsy showed necrotizing fungal pneumonia with negative culture. The dosage of LAMB was increased, and itraconazole (ITRA) was added; subsequently LAMB was discontinued and therapy was continued with ITRA alone. The patient demonstrated clinical, and radiological improvement. In vitro, the isolate was susceptible to low concentrations of AMB and ITRA. A. fusispora is a thermotolerant, fast-growing fungus with neurotropic potential. We report the first case of human infection involving the CNS. Acrophialophora resembles Paecilomyces but differs in having colonies that become dark and in the development of phialides along the sides or at the tips of echinulate brown conidiophores. Conidia are horne in long chains and are smooth or ornamented with fine-to coarse echinulations, sometimes in spiral bands. The taxonomy of the genus Acrophialophora is reviewed, and Acrophialophora nainiana and Acrophialophora levis are considered as synonyms of A. fusispora. C1 King Faisal Specialist Hosp & Res Ctr, Dept Pediat, Riyadh 11211, Saudi Arabia. Univ Texas, Hlth Sci Ctr, Fungus Testing Lab, San Antonio, TX USA. Univ Alberta, Microfungus Collect, Edmonton, AB, Canada. NCI, Pediat Branch, NIH, Bethesda, MD 20892 USA. RP Al-Mohsen, IZ (reprint author), King Faisal Specialist Hosp & Res Ctr, Dept Pediat, POB 3354, Riyadh 11211, Saudi Arabia. NR 29 TC 19 Z9 19 U1 1 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0095-1137 J9 J CLIN MICROBIOL JI J. Clin. Microbiol. PD DEC PY 2000 VL 38 IS 12 BP 4569 EP 4576 PG 8 WC Microbiology SC Microbiology GA 399ND UT WOS:000166818500043 PM 11101597 ER PT J AU Grem, JL Harold, N Shapiro, J Bi, DQ Quinn, MG Zentko, S Keith, B Hamilton, JM Monahan, BP Donavan, S Grollman, F Morrison, G Takimoto, CH AF Grem, JL Harold, N Shapiro, J Bi, DQ Quinn, MG Zentko, S Keith, B Hamilton, JM Monahan, BP Donavan, S Grollman, F Morrison, G Takimoto, CH TI Phase I and pharmacokinetic trial of weekly oral fluorouracil given with eniluracil and low-dose leucovorin to patients with solid tumors SO JOURNAL OF CLINICAL ONCOLOGY LA English DT Article ID DIHYDROPYRIMIDINE DEHYDROGENASE-ACTIVITY; ADVANCED COLORECTAL-CANCER; 5-ETHYNYLURACIL 776C85; INFUSION FLUOROURACIL; 24-HOUR INFUSION; RANDOMIZED TRIAL; BETA-ALANINE; 5-FLUOROURACIL; MODULATION; INACTIVATION AB Purpose: Fluorouracil (5-FU) given as a weekly, high-dose 24-hour infusion is active and tolerable. We evaluated an oral regimen of eniluracil (which inactivates dihydropyrimidine dehydrogenase [DPD]), 5-FU, and leucovorin ta simulate this schedule. Patients and Methods: Patients received a single 24-hour infusion of 5-FU (2,300 mg/m(2) on day 2) with leucovorin(15 mg orally [PO] bid on days 1 through 3) to provide reference pharmacokinetic data. Two weeks later, patients began treatment with eniluracil (20 mg) and leucovorin (15 mg) (90 bid on days 1 through 3) and 5-FU (10 to 15 mg/m2 PO bid on day 2). Results: Dose-limiting toxicity (diarrhea, neutropenia, and fatigue) was seen with 5-FU 15 mg/m2 PO bid on day 2 given weekly for either 6 of 8 weeks or 3 of 4 weeks, whereas five of seven patients tolerated 5-FU 10 mg/m2 PO bid given weekly for 3 of 4 weeks. Eniluracil led to a 35-fold reduction in 5-FU clearance. Fluoro-beta-alanine, a 5-FU catabolite, was not detected in plasma during oral 5-FU-eniluracil therapy. DPD activity was markedly suppressed in all patients during eniluracil therapy; the inactivation persisted after the last eniluracil dose; percentages of baseline values were 1.8% on day 5, 4.5% on day 12, and 23.6% on day 19. Conclusion: The recommended oral dosage of 5-FU (10 mg/m2 PO bid) given with eniluracil and leucovorin is approximately 115-fold lower than the reference dosage for 24-hour infusional 5-FU. This difference is greater than expected given the reduction in 5-FU clearance. DPD inactivation persisted for several weeks after completion of eniluracil therapy. (C) 2000 by American Society of Clinical Oncology. C1 NCI, Med Branch, Natl Naval Med Ctr, Div Clin Sci, Bethesda, MD 20889 USA. Natl Naval Med Ctr, Dept Internal Med, Hematol Oncol Sect, Bethesda, MD USA. Natl Naval Med Ctr, Dept Radiol, Bethesda, MD USA. RP Grem, JL (reprint author), NCI, Med Branch, Natl Naval Med Ctr, Div Clin Sci, 8901 Wisconsin Ave,Bldg 8,Rm 5101, Bethesda, MD 20889 USA. NR 49 TC 17 Z9 17 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0732-183X J9 J CLIN ONCOL JI J. Clin. Oncol. PD DEC 1 PY 2000 VL 18 IS 23 BP 3952 EP 3963 PG 12 WC Oncology SC Oncology GA 380UJ UT WOS:000165721600010 PM 11099325 ER PT J AU Marshall, JL Hoyer, RJ Toomey, MA Faraguna, K Chang, P Richmond, E Pedicano, JE Gehan, E Peck, RA Arlen, P Tsang, KY Schlom, J AF Marshall, JL Hoyer, RJ Toomey, MA Faraguna, K Chang, P Richmond, E Pedicano, JE Gehan, E Peck, RA Arlen, P Tsang, KY Schlom, J TI Phase I study in advanced cancer patients of a diversified prime-and-boost vaccination protocol using recombinant vaccinia virus and recombinant nonreplicating avipox virus to elicit anti-carcinoembryonic antigen immune responses SO JOURNAL OF CLINICAL ONCOLOGY LA English DT Article ID CYTOTOXIC T-LYMPHOCYTE; STIMULATING FACTOR; EPITOPES; PEPTIDE; CEA; IDENTIFICATION; CARCINOMA; ASSAY; SERUM; CELLS AB Purpose: This trial sought to determine, for the first time, the validity in human vaccinations of using two different recombinant vaccines in diversified prime-and-boost regimens to enhance T-cell responses to a tumor antigen. Patients and Methods: Eighteen patients with advanced tumors expressing carcinoembryonic antigen (CEA) were randomized to receive either recombinant vaccinia (rV)-CEA followed by three avipox-CEA vaccinations, or avipox-CEA (three times) followed by one rV-CEA vaccination. Subsequent vaccinations in both cohorts were with avipox-CEA. Immunologic monitoring was performed using a CEA peptide and the enzyme-linked immunospot assay for interferon gamma production. Results: rV-CEA followed by avipox-CEA was superior ta the reverse order in the generation of CFA-specific T-cell responses. Further increases in CFA-specific T-cell precursors were seen when local granulocyte-macrophage col-any-stimulating factor (GM-CSF) and low-dose interleukin (IL)-2 were given with subsequent vaccinations. The treatment was extremely well tolerated. Limited clinical activity was seen using vaccines alone in this patient population. Antibody production against CEA was also observed in some of the treated patients. Conclusion: rV-CEA was more effective in its role as a primer of the immune system; avipox-CEA could be given up to eight times with continued increases in CEA T-cell precursors. Future trials should use rV-CEA first followed by avipox-CEA. Vaccines specific to CEA are able to generate CEA-specific T-cell responses in patients without significant toxicity. T-cell responses using vaccines alone may be inadequate to generate significant anticancer objective responses in patients with advanced disease. Cytokines such as GM-CSF and IL-2 may play a key role in generating such responses. (C) 2000 by American Society of Clinical Oncology. C1 Georgetown Univ, Med Ctr, Vincent T Lombardi Canc Ctr, Washington, DC 20007 USA. NCI, Div Basic Sci, Tumor Immunol & Biol Lab, NIH, Bethesda, MD 20892 USA. Univ Virginia, Hlth Sci Ctr, Charlottesville, VA USA. RP Marshall, JL (reprint author), Georgetown Univ, Med Ctr, Vincent T Lombardi Canc Ctr, 3800 Reservoir Rd NW, Washington, DC 20007 USA. NR 30 TC 258 Z9 267 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0732-183X J9 J CLIN ONCOL JI J. Clin. Oncol. PD DEC 1 PY 2000 VL 18 IS 23 BP 3964 EP 3973 PG 10 WC Oncology SC Oncology GA 380UJ UT WOS:000165721600011 PM 11099326 ER PT J AU Frye, MA Ketter, TA Kimbrell, TA Dunn, RT Speer, AM Osuch, EA Luckenbaugh, DA Cora-Locatelli, G Leverich, GS Post, RM AF Frye, MA Ketter, TA Kimbrell, TA Dunn, RT Speer, AM Osuch, EA Luckenbaugh, DA Cora-Locatelli, G Leverich, GS Post, RM TI A placebo-controlled study of lamotrigine and gabapentin monotherapy in refractory mood disorders SO JOURNAL OF CLINICAL PSYCHOPHARMACOLOGY LA English DT Article; Proceedings Paper CT 151st Annual Meeting of the American-Psychiatric-Association CY MAY 30-JUN 05, 1998 CL TORONTO, CANADA SP Amer Psychiat Assoc ID CYCLING BIPOLAR DISORDER; DOUBLE-BLIND; ANTIEPILEPTIC DRUG; KINDLED SEIZURES; ACUTE MANIA; LITHIUM; DEPRESSION; EFFICACY; ILLNESS; CARBAMAZEPINE AB There is a pressing need for additional treatment options for refractory mood disorders. This controlled comparative study evaluated the efficacy of lamotrigine (LTG) and gabapentin (GBP) monotherapy versus placebo (PLC). Thirty-one patients with refractory bipolar and unipolar mood disorders participated in a double-blind, randomized, crossover series of three 6-week monotherapy evaluations including LTG, GBP, and PLC. There was a standardized blinded titration to assess clinical efficacy or to determine the maximum tolerated daily dose (LTG 500 mg or GBP 4,800 mg). The primary outcome measure was the Clinical Global Impressions Scale (CGI) for Bipolar Illness as supplemented by other standard rating instruments. The mean doses at week 6 were 274 +/- 128 mg for LTG and 3,987 +/- 856 mg for GBP. Response rates (CGI ratings of much or very much improved) were the following: LTG, 52% (16/31); GBP, 26% (8/31); and PLC, 23% (7/31) (Cochran's Q = 6.952, df = 2, N = 31, p = 0.031). Post hoc Q differences (df = 1, N = 31) were the following: LTG versus GBP (Q, 5.33, p = 0.011); LTG versus PLC (Q(diff) = 4.76, p = 0.022); and GBP versus PLC (Q(diff) = 0.08,p = 0.70). With respect to anticonvulsant dose and gender, there was no difference between the responders and the nonresponders. The agents mere generally well tolerated. This controlled investigation preliminarily suggests the efficacy of LTG in treatment-refractory affectively ill patients. Further definition of responsive subtypes and the role of these medications in the treatment of mood disorders requires additional study. C1 NIMH, Biol Psychiat Branch, NIH, Bethesda, MD 20892 USA. NIMH, Clin Sci Lab, NIH, Bethesda, MD 20892 USA. Univ Calif Los Angeles, Sch Med, Los Angeles, CA USA. Stanford Univ, Sch Med, Stanford, CA 94305 USA. Univ Arkansas, Sch Med, Little Rock, AR 72204 USA. RP Post, RM (reprint author), NIMH, Biol Psychiat Branch, NIH, Bldg 10,Room 3N212,10 Ctr Dr,MSC 1272, Bethesda, MD 20892 USA. RI Osuch, Elizabeth/B-5009-2015 OI Osuch, Elizabeth/0000-0001-5946-1862 NR 62 TC 318 Z9 327 U1 1 U2 12 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0271-0749 J9 J CLIN PSYCHOPHARM JI J. Clin. Psychopharmacol. PD DEC PY 2000 VL 20 IS 6 BP 607 EP 614 DI 10.1097/00004714-200012000-00004 PG 8 WC Pharmacology & Pharmacy; Psychiatry SC Pharmacology & Pharmacy; Psychiatry GA 377AU UT WOS:000165490400003 PM 11106131 ER PT J AU Nahas, Z DeBrux, C Chandler, V Lorberbaum, JP Speer, AM Molloy, MA Liberatos, C Risch, SC George, MS AF Nahas, Z DeBrux, C Chandler, V Lorberbaum, JP Speer, AM Molloy, MA Liberatos, C Risch, SC George, MS TI Lack of significant changes on magnetic resonance scans before and after 2 weeks of daily left prefrontal repetitive transcranial magnetic stimulation for depression SO JOURNAL OF ECT LA English DT Article DE depression; repetitive transcranial magnetic stimulation; magnetic resonance imaging ID MAJOR DEPRESSION; HEARING-LOSS; CORTEX; BRAIN; RTMS; MOOD; SAFETY AB Repetitive transcranial magnetic stimulation (rTMS) is a new technology for exploring brain function. With this method, a small electromagnet is placed on the scalp; by activating and deactivating it, nerve cells in the underlying superficial cortex are depolarized. Several studies have found that prefrontal rTMS has potential efficacy in treating depression, and this technology, in addition to being a research tool, may soon play a role in psychiatric practice. Thus, establishing the safety of this technology is important and has been studied insufficiently. The authors performed T1-weighted three-dimensional volumetric magnetic resonance (MR) imaging on 22 depressed adults (15 active, 7 control) before and after they participated in a 2-week double-blinded, placebo-controlled trial of daily left prefrontal rTMS for the treatment of depression (a total of 16,000 stimuli). Seventeen patients also had paired T2-weighted scans. In a blinded manner, MR scans were qualitatively and quantitatively assessed for structural changes. No qualitative structural differences were observed before and after treatment. In addition, volumetric analysis of the prefrontal lobe showed no changes in the 2 weeks of the study. In conclusion, 10 days of daily prefrontal rTMS at these intensities and frequencies does not cause observable structural changes on MR scans in depressed adults. C1 Med Univ S Carolina, Inst Psychiat, Dept Psychiat, Charleston, SC 29425 USA. Med Univ S Carolina, Dept Radiol, Charleston, SC 29425 USA. Med Univ S Carolina, Dept Neurol, Charleston, SC 29425 USA. NIMH, Biol Psychiat Branch, Bethesda, MD 20892 USA. Ralph H Johnson Vet Adm Med Ctr, Dept Psychiat, Charleston, SC USA. RP Nahas, Z (reprint author), Med Univ S Carolina, Inst Psychiat, Dept Psychiat, 67 President St, Charleston, SC 29425 USA. NR 41 TC 40 Z9 40 U1 1 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1095-0680 J9 J ECT JI J. ECT PD DEC PY 2000 VL 16 IS 4 BP 380 EP 390 DI 10.1097/00124509-200012000-00008 PG 11 WC Behavioral Sciences; Psychiatry SC Behavioral Sciences; Psychiatry GA 382HC UT WOS:000165816000008 PM 11314876 ER PT J AU Vineis, P Miligi, L Crosignani, P Fontana, A Masala, G Nanni, O Ramazzotti, V Rodella, S Stagnaro, E Tumino, R Vigano, C Vindigni, C Costantini, AS AF Vineis, P Miligi, L Crosignani, P Fontana, A Masala, G Nanni, O Ramazzotti, V Rodella, S Stagnaro, E Tumino, R Vigano, C Vindigni, C Costantini, AS TI Delayed infection, family size and malignant lymphomas SO JOURNAL OF EPIDEMIOLOGY AND COMMUNITY HEALTH LA English DT Article ID NON-HODGKINS-LYMPHOMA; DAY-CARE; ASSOCIATION; RESPONSES; DISEASES AB Background-The annual incidence of non-Hodgkin's lymphomas (NHL) is increasing by 3%-4% in different parts of the developed world. Excesses of NHL have been observed in populations exposed to immunosuppressants and to HIV, but these causes do not explain the increasing trends. It is suggested that delayed infection could explain NHL trends, through an impairment of the Th1/Th2 lymphocyte patterns. Methods-In a population-based study on 1388 patients with NHL, 354 with Hodgkin's disease (HD) and 1718 healthy controls, the age of first occurrence of bacterial and viral diseases was investigated. Clinical records were perused in one centre to check the anamnestic data. Findings-The age of occurrence of bacterial and viral diseases was significantly higher among NHL patients than in the controls. The association between later age at first bacterial or viral disease was limited to small families (OR= 1.95; 95% confidence intervals 1.26, 3.00, for age 4-8 at first infection; OR=1.91; 1.19, 3.06, for age 9+, compared with less than 4). The association was more obvious for bacterial diseases (possibly for the lower degree of misclassification). High grade lymphomas showed the strongest association. The later age of occurrence of bacterial or viral diseases in NHL patients is consistent with a higher incidence of lymphomas observed in higher social groups. No clear association was found between HD and age at first bacterial or viral diseases. Interpretation-It is proposed that delayed infection could explain the increasing NHL trends, through an impairment of the Th1/Th2 lymphocyte patterns. The model of delayed infection has been proposed also to explain increasing prevalence rates of asthma. C1 Osped S Giovanni Battista, Serv Epidemiol Tumori, I-10123 Turin, Italy. Univ Turin, I-10123 Turin, Italy. Ctr Studio Prevenz Oncol, Florence, Italy. Natl Canc Inst, I-20133 Milan, Italy. Local Hlth Unit, Novara, Italy. Ist Oncol Romagnolo, Forli, Italy. Canc Registry, Latina, Italy. Local Hlth Unit, Verona, Italy. Natl Canc Inst, Genoa, Italy. Canc Registry, Ragusa, Italy. Univ Siena, Dept Pathol, I-53100 Siena, Italy. RP Vineis, P (reprint author), Osped S Giovanni Battista, Serv Epidemiol Tumori, Via Santena 7, I-10123 Turin, Italy. OI NANNI, ORIANA/0000-0001-7338-2896 FU NCI NIH HHS [CA51086] NR 34 TC 43 Z9 43 U1 0 U2 1 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 0143-005X J9 J EPIDEMIOL COMMUN H JI J. Epidemiol. Community Health PD DEC PY 2000 VL 54 IS 12 BP 907 EP 911 DI 10.1136/jech.54.12.907 PG 5 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 374LE UT WOS:000165346000007 PM 11076986 ER PT J AU Halabi, S Skinner, CS Samsa, GP Strigo, TS Crawford, YS Rimer, BK AF Halabi, S Skinner, CS Samsa, GP Strigo, TS Crawford, YS Rimer, BK TI Factors associated with repeat mammography screening SO JOURNAL OF FAMILY PRACTICE LA English DT Article DE mammography; breast; repeat screening [non-MESH]; vaginal smears; hormone replacement therapy [non-MESH] ID OLDER; WOMEN; POPULATION; PATTERNS; STAGE AB BACKGROUND Even organizations with differing mammography recommendations agree that regular repeat screening is required for mortality reduction. However, most studies have focused on one-time screening rather than repeat adherence. We compare trends in beliefs and health-related behaviors among women screened and adherent to the National Cancer Institute's screening mammography recommendations (on schedule), those screened at least once and nonadherent (off schedule), and those never screened, METHODS Our data are from a baseline telephone interview conducted among 1287 female members of Blue Cross Blue Shield of North Carolina who were aged either 40 to 44 years or 50 to 54 years. RESULTS The 3 groups differed significantly on beliefs and health-related behaviors, with the off-schedule group almost consistently falling bem een the on-schedule and never screened groups. Off-schedule women were more likely than on-schedule women, but less likely than those never screened, to not have a clinical breast examination within 12 months, to be ambivalent about screening mammography, to be confused about screening guidelines, and to not be advised by a physician to get a mammogram in the past 2 years. Off-schedule women perceived their breast cancer risk as lower and were less likely to be up to date with other cancer screening tests. CONCLUSIONS Our findings suggest that women who are off schedule are in need of mammography-promoting interventions, including recommendations from and discussion with their health care providers, Because they are more positive and knowledgeable about mammography than women who have never been screened they may benefit from brief interventions from health care providers that highlight the importance of repeat screening. C1 Duke Univ, Ctr Med, Dept Community & Family Med, Div Biometry, Durham, NC 27710 USA. Duke Univ, Med Ctr, Duke Comprehens Canc Ctr, Canc Prevent Detect & Control Res Program, Durham, NC 27710 USA. NCI, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. RP Halabi, S (reprint author), Duke Univ, Ctr Med, Dept Community & Family Med, Div Biometry, Box 3958, Durham, NC 27710 USA. FU NCI NIH HHS [5U19-CA-72099-03] NR 33 TC 62 Z9 62 U1 2 U2 3 PU DOWDEN PUBLISHING CORP PI MONTVALE PA 110 SUMMIT AVE, MONTVALE, NJ 07645-1712 USA SN 0094-3509 J9 J FAM PRACTICE JI J. Fam. Pract. PD DEC PY 2000 VL 49 IS 12 BP 1104 EP 1112 PG 9 WC Primary Health Care; Medicine, General & Internal SC General & Internal Medicine GA 381DY UT WOS:000165748100006 PM 11132060 ER PT J AU Niemela, O Parkkila, S Juvonen, RO Viitala, K Gelboin, HV Pasanen, M AF Niemela, O Parkkila, S Juvonen, RO Viitala, K Gelboin, HV Pasanen, M TI Cytochromes P450 2A6 2E1, and 3A and production of protein-aldehyde adducts in the liver of patients with alcoholic and non-alcoholic liver diseases SO JOURNAL OF HEPATOLOGY LA English DT Article DE diabetes; ethanol; hepatitis; hepatotoxicity; microsomal metabolism ID ACETALDEHYDE-MODIFIED EPITOPES; ETHANOL OXIDIZING SYSTEM; LOW-DENSITY-LIPOPROTEIN; LIPID-PEROXIDATION; MONOCLONAL-ANTIBODIES; CYP ENZYMES; EXPRESSION; METABOLISM; INJURY; MOUSE AB Background/Aims: Interaction between CYP2E1, ethanol metabolites, and enhanced lipid peroxidation is linked to the pathogenesis of alcoholic liver disease. This study was conducted to compare the expression of various cytochrome enzymes and the appearance of aldehyde adducts in humans. Methods: Acetaldehyde- and lipid peroxidation-derived protein adducts and CYP2A6, 2E1, and 3A4/5 were examined immunohistochemically from liver specimens of 12 alcohol abusers with either mild (n=7) or severe (n=5) liver disease, and from nine nondrinking patients with non-alcoholic steatosis (n=4), or hepatitis (n=5). Results: Ethanol-inducible CYP2E1 was present in all alcoholic livers. While CYP2A6 in zone 3 hepatocytes was also abundant in the alcoholic patients with various degrees of liver disease, CYP3A4/5 was most prominent in alcoholic cirrhosis, The sites of CYP2E1 and CYP2A6 immunoreactivity co-localized with fatty deposits, and with the sites of acetaldehyde and lipid peroxidation-derived protein adducts, The CYP enzymes were also abundant in the centrilobular hepatocytes of patients,vith fatty liver due to obesity or diabetes. Conclusions: Alcohol-induced liver damage is associated with a generalized induction of CYP2A6, CYP2E1 and CYP3A4 and generation of acetaldehyde and lipid peroxidation-derived protein-aldehyde adducts, However, CYP induction also occurred in patients with non-alcoholic steatosis. C1 Oulu Univ, Dept Clin Chem, FIN-90401 Oulu, Finland. EP Cent Hosp Lab, Seinajoki, Finland. Univ Kuopio, Dept Pharmacol, FIN-70211 Kuopio, Finland. NCI, Bethesda, MD 20892 USA. Natl Agcy Med, Helsinki, Finland. RP Pasanen, M (reprint author), Oulu Univ, Dept Pharmacol, PL 5000, FIN-90401 Oulu, Finland. NR 51 TC 82 Z9 97 U1 1 U2 8 PU MUNKSGAARD INT PUBL LTD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0168-8278 J9 J HEPATOL JI J. Hepatol. PD DEC PY 2000 VL 33 IS 6 BP 893 EP 901 DI 10.1016/S0168-8278(00)80120-8 PG 9 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 378QU UT WOS:000165595700005 PM 11131450 ER PT J AU St John, PL Robert, B Wang, RX Abrahamson, DR AF St John, PL Robert, B Wang, RX Abrahamson, DR TI Abnormal laminin synthesis by organ cultured fetal kidneys is corrected by grafting. SO JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY LA English DT Meeting Abstract C1 Univ Kansas, Med Ctr, Lawrence, KS 66045 USA. NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU HISTOCHEMICAL SOC INC PI SEATTLE PA UNIV WASHINGTON, DEPT BIOSTRUCTURE, BOX 357420, SEATTLE, WA 98195 USA SN 0022-1554 J9 J HISTOCHEM CYTOCHEM JI J. Histochem. Cytochem. PD DEC PY 2000 VL 48 IS 12 MA 32 BP 1728 EP 1728 PG 1 WC Cell Biology SC Cell Biology GA 384BV UT WOS:000165919900046 ER PT J AU Muraro, PA Jacobsen, M Necker, A Nagle, JW Gaber, R Sommer, N Oertel, WH Martin, R Hemmer, B AF Muraro, PA Jacobsen, M Necker, A Nagle, JW Gaber, R Sommer, N Oertel, WH Martin, R Hemmer, B TI Rapid identification of local T cell expansion in inflammatory organ diseases by flow cytometric T cell receptor V beta analysis SO JOURNAL OF IMMUNOLOGICAL METHODS LA English DT Article DE T cell receptors; flow cytometry; monoclonal antibodies; cerebrospinal fluid; encephalitis ID TUMOR-INFILTRATING LYMPHOCYTES; PRIMARY IMMUNE-RESPONSE; RHEUMATOID-ARTHRITIS; AUTOIMMUNE-DISEASES; HEALTHY-INDIVIDUALS; MULTIPLE-SCLEROSIS; CLONAL EXPANSIONS; REPERTOIRE; EXPRESSION; GENES AB Oligoclonal expansion of antigen-specific T cells occurs frequently during inflammatory diseases. These cells may persist for a long time at high frequency in the body and be enriched in the affected tissues. As a screening test for expanded cell T cell populations at sites of inflammation, we developed an optimized methodology for flow-cytometry-based quantification of T cell receptor V beta (TCRBV) expression. We first validated the specificity of a TCRBV-specific monoclonal antibody set by direct comparison with PCR-based analysis of mono- and polyclonal T cell samples. This monoclonal antibody (mAb) panel recognized approximately two thirds of the T cell receptor alpha/beta repertoire in agroup of 64 healthy donors and allowed defining TCR usage in the CD4+ and CD8+ subsets. The reliable detection of expanded V beta gene families in T cell populations was confirmed in experiments on superantigen-stimulated T cells. Through differential TCR analysis on T cell subpopulations in cerebrospinal fluid and blood in patients with acute encephalitis, we were able to identify locally expanded CD8+ T cells. The power of this approach affords not only high-throughput comparative TCR analysis for immunological studies in vitro, but also rapid ex vivo identification of cell populations enriched in organ compartments during inflammatory diseases. (C) 2000 Elsevier Science B.V; All rights reserved. C1 Univ G DAnnunzio, Med Sch,Neurol Clin, Osped Clin, Dept Oncol & Neurosci, I-66013 Chieti, Italy. NINDS, Neuroimmunol Branch, NIH, Bethesda, MD 20892 USA. Univ Marburg, Dept Neurol, Clin Neuroimmunol Grp, Marburg, Germany. Immunotech, Antibody Dept, Marseille, France. NINDS, DNA Sequencing Facil, NIH, Bethesda, MD 20892 USA. RP Muraro, PA (reprint author), Univ G DAnnunzio, Med Sch,Neurol Clin, Osped Clin, Dept Oncol & Neurosci, Via Vestini, I-66013 Chieti, Italy. OI Muraro, Paolo/0000-0002-3822-1218 NR 50 TC 38 Z9 38 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0022-1759 J9 J IMMUNOL METHODS JI J. Immunol. Methods PD DEC 1 PY 2000 VL 246 IS 1-2 BP 131 EP 143 DI 10.1016/S0022-1759(00)00309-4 PG 13 WC Biochemical Research Methods; Immunology SC Biochemistry & Molecular Biology; Immunology GA 380JW UT WOS:000165699300013 PM 11121554 ER PT J AU Sproul, TW Malapati, S Kim, J Pierce, SK AF Sproul, TW Malapati, S Kim, J Pierce, SK TI Cutting edge: B cell antigen receptor signaling occurs outside lipid rafts in immature B cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID TRANSDUCTION; ACTIVATION; APOPTOSIS; LYMPHOMAS; TOLERANCE; BCR AB B cell Ag receptor (BCR) signaling changes dramatically during B cell development, resulting in activation in mature B cells and apoptosis, receptor editing, or anergy in immature Ei cells. BCR signaling in mature B cells was shown to be initiated by the translocation of the BCR into cholesterol- and sphingolipid-enriched membrane microdomains that include the Src family kinase Lyn and exclude the phosphatase CD45. Subsequently the BCR is rapidly internalized into the cell. Here we show that the BCR in the immature B cell line, WEHI-231, does not translocate into lipid rafts following cross-linking nor is the BCR rapidly internalized, The immature BCR initiates signaling from outside lipid rafts as evidenced by the immediate induction of an array of phosphoproteins and subsequent apoptosis, The failure of the BCR in immature B cells to enter lipid rafts may Contribute to the dramatic difference in the outcome of signaling in mature and immature B cells. C1 NIAID, Immunogenet Lab, NIH, Rockville, MD 20852 USA. Northwestern Univ, Dept Biochem Mol Biol & Cell Biol, Evanston, IL 60208 USA. RP Pierce, SK (reprint author), NIAID, Immunogenet Lab, NIH, Twinbrook 2,12441 Parklawn Dr,Room 200B,MSC 8180, Rockville, MD 20852 USA. FU NIAID NIH HHS [AI 27957, AI40309, AI18939] NR 18 TC 87 Z9 89 U1 0 U2 0 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD DEC 1 PY 2000 VL 165 IS 11 BP 6020 EP 6023 PG 4 WC Immunology SC Immunology GA 376PZ UT WOS:000165467100002 PM 11086032 ER PT J AU Ostrowski, MA Justement, SJ Ehler, L Mizell, SB Lui, SY Mican, J Walker, BD Thomas, EK Seder, R Fauci, AS AF Ostrowski, MA Justement, SJ Ehler, L Mizell, SB Lui, SY Mican, J Walker, BD Thomas, EK Seder, R Fauci, AS TI The role of CD4(+) T cell help and CD40 ligand in the in vitro expansion of HIV-1-specific memory cytotoxic CD8(+) T cell responses SO JOURNAL OF IMMUNOLOGY LA English DT Article ID ACQUIRED IMMUNODEFICIENCY SYNDROME; PRIMARY IMMUNE-RESPONSE; MHC CLASS-II; DENDRITIC CELLS; LYMPHOCYTE RESPONSES; HIV-1 INFECTION; SOLUBLE-ANTIGEN; VIRUS-INFECTION; PLASMA VIREMIA; ACTIVATION AB CD4(+) T cells have been shown to play a critical role in the maintenance of an effective anti-viral CD8(+) CTL response in murine models. Recent studies have demonstrated that CD4(+) T cells provide help to CTLs through ligation of the CD40 receptor on dendritic cells, The role of CD4(+) T cell help in the expansion of virus-specific CD8(+) memory T cell responses was examined in normal volunteers recently vaccinated to influenza and in HIV-1 infected individuals. In recently vaccinated normal volunteers, CD4(+) T cell help was required for optimal in vitro expansion of influenza specific CTL responses, Also, CD40 ligand trimer (CD40LT) enhanced CTL responses and was able to completely substitute for CD4(+) T cell help in PBMCs from normal volunteers. In HIV-1 infection, CD4(+) T cell help was required for optimal expansion of HIV-l-specific memory CTL in vitro in 9 of 10 patients. CD40LT could enhance CTL in the absence of CD4(+) T cell help in the majority of patients; however, the degree of enhancement of CTL responses was variable such that, in some patients, CD40LT could not completely substitute for CD4(+) T cell help. In those HIV-l-infected patients who demonstrated poor responses to CD40LT, a dysfunction in circulating CD8(+) memory T cells was demonstrated, which was reversed by the addition of cytokines including IL-2, Finally, it was demonstrated that IL-15 produced by CD40LT-stimulated dendritic cells may be an additional mechanism by which CD40LT induces the expansion of memory CTL in CD4(+) T cell-depleted conditions, where IL-2 is lacking. C1 NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. Massachusetts Gen Hosp, Partners AIDS Res Ctr, Charlestown, MA 02129 USA. Massachusetts Gen Hosp, Div Infect Dis, Charlestown, MA 02129 USA. Harvard Univ, Sch Med, Charlestown, MA 02129 USA. Immunex Res & Dev Corp, Seattle, WA 98101 USA. RP Ostrowski, MA (reprint author), Univ Toronto, Div Clin Sci, Room 6271,1 Kings Coll Circle, Toronto, ON M5S 1A8, Canada. EM m.ostrowski@utoronto.ca NR 51 TC 80 Z9 83 U1 0 U2 2 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD DEC 1 PY 2000 VL 165 IS 11 BP 6133 EP 6141 PG 9 WC Immunology SC Immunology GA 376PZ UT WOS:000165467100016 PM 11086046 ER PT J AU Wu, CY Wang, X Gadina, M O'Shea, JJ Presky, DH Magram, J AF Wu, CY Wang, X Gadina, M O'Shea, JJ Presky, DH Magram, J TI IL-12 receptor beta 2 (IL-12R beta 2)-deficient mice are defective in IL-12-mediated signaling despite the presence of high affinity IL-12 binding sites SO JOURNAL OF IMMUNOLOGY LA English DT Article ID ACTIVATED HUMAN LYMPHOBLASTS; STEM-CELL LINES; CD4+ T-CELLS; INTERFERON-GAMMA; DEFICIENT MICE; TH2 CELLS; IN-VITRO; INTERLEUKIN-12; EXPRESSION; COMPONENT AB Two subunits of the IL-12 receptor (IL-12R), IL-12R beta1 and IL-12R beta2, have been identified and cloned. Previous studied dem demonstrated that the IL-12R beta1 subunit was required for mouse T and NK cells to respond to IL-12 in vivo. To investigate the role of IL-12R beta2 in IL-12 signaling, we have generated IL-12R beta2-deficient (IL-12R beta2(-/-)) mice by targeted mutation in embryonic stem (ES) cells, Although Con A-activated splenocytes from IL-12R beta2(-/-) mice still bind IL-12 with both high and low affinity, no IL-12-induced biological functions can be detected, Con A-activated splenocytes of IL-12R beta2(-/-) mice failed to produce IFN-gamma or proliferate in response to IL-12 stimulation. NK lytic activity of IL-12R beta2(-/-) splenocytes was not induced when incubated with IL-12, IL-12R beta2(-/-) splenocytes were deficient in IFN-gamma secretion when stimulated with either Con A or anti-CD3 mAb in vitro, Furthermore, IL-12R beta2(-/-) mice were deficient in vivo in their ability to produce IFN-gamma following endotoxin administration and to generate a type I cytokine response, IL-12-mediated signal transduction was also defective as measured by phosphorylation of STAT4, These results demonstrate that although mouse IL-12R beta1 is the subunit primarily responsible for binding IL-12, IL-12R beta2 plays an essential role in mediating the biological functions of IL-12 in mice. C1 Hoffmann La Roche Inc, Dept Inflammat Autoimmune Dis, Nutley, NJ 07110 USA. NIAMSD, Lymphocyte Cell Biol Sect, Arthrit & Rheumatism Branch, NIH, Bethesda, MD 20892 USA. RP Magram, J (reprint author), OSI Pharmaceut, 777 Old Saw Mill River Rd, Tarrytown, NY 10591 USA. NR 39 TC 115 Z9 129 U1 0 U2 1 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD DEC 1 PY 2000 VL 165 IS 11 BP 6221 EP 6228 PG 8 WC Immunology SC Immunology GA 376PZ UT WOS:000165467100026 PM 11086056 ER PT J AU Lee, JS Monson, NL Lipsky, PE AF Lee, JS Monson, NL Lipsky, PE TI The V lambda J lambda repertoire in human fetal spleen: Evidence for positive selection and extensive receptor editing SO JOURNAL OF IMMUNOLOGY LA English DT Article ID PERIPHERAL B-CELLS; JUNCTIONAL DIVERSITY; GENE REPERTOIRE; PRE-B; PREFERENTIAL UTILIZATION; MOLECULAR MECHANISMS; NEGATIVE SELECTION; LYMPHOCYTES-B; CO-RECEPTORS; EXPRESSION AB V lambdaJ lambda rearrangements obtained from genomic DNA of individual IgM(+) B cells from human fetal spleen were analyzed. A nonrandom pattern of lambda gene rearrangements that differed from the adult VA repertoire was found. The VA distal genes 8A and 4B were absent from the nonproductive fetal repertoire, whereas 2E and 3L were overrepresented;and 1B was underrepresented in the productive fetal repertoire. Positive selection of the V zeta gene, 2E, along with V lambda rearrangements employing homologous V lambdaJ lambda joins were observed in the fetal, but not in the adult V lambda repertoire. Overrepresentation of J lambda distal cluster C genes rearranging to the V lambda distal J segment, J lambda7, in both productive and nonproductive fetal repertoires suggested that receptor editing/replacement was more active in the fetus than in adults. Numerous identical V lambdaJ lambda junctions were observed in both the productive and nonproductive repertoire of the fetus and adult, but were significantly more frequent in the productive repertoire of the fetus, suggesting expansion of B cells expressing particular lambda -light chains in both stages of development, with more profound expansion in the fetal repertoire. Notably, B cells expressing identical lambda -light chains expressed diverse heavy chains. These data demonstrate that three mechanisms strongly influence the shaping of the human fetal lambda -chain repertoire that are less evident in the adult: positive selection, receptor editing, and expansion of B cells expressing specific lambda -light chains. These events imply that the expressed fetal repertoire is shaped by exposure to self Ags. C1 Univ Texas, SW Med Ctr, Dept Internal Med, Dallas, TX 75235 USA. Univ Texas, SW Med Ctr, Harold C Simmons Arthritis Res Ctr, Dallas, TX 75235 USA. RP Lipsky, PE (reprint author), NIAMSD, Autoimmun Branch, NIH, 10 Ctr Dr,MSC 1820, Bethesda, MD 20892 USA. NR 55 TC 25 Z9 25 U1 0 U2 1 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD DEC 1 PY 2000 VL 165 IS 11 BP 6322 EP 6333 PG 12 WC Immunology SC Immunology GA 376PZ UT WOS:000165467100039 PM 11086069 ER PT J AU Rowe, JA Rogerson, SJ Raza, A Moulds, JM Kazatchkine, MD Marsh, K Newbold, CI Atkinson, JP Miller, LH AF Rowe, JA Rogerson, SJ Raza, A Moulds, JM Kazatchkine, MD Marsh, K Newbold, CI Atkinson, JP Miller, LH TI Mapping of the region of complement receptor (CR) 1 required for Plasmodium falciparum resetting and demonstration of the importance of CR1 in resetting in field isolates SO JOURNAL OF IMMUNOLOGY LA English DT Article ID INFECTED ERYTHROCYTES; MONOCLONAL-ANTIBODIES; HUMAN MONOCYTES; C3B RECEPTOR; ACTIVE-SITES; MALARIA; CD35; IDENTIFICATION; PHAGOCYTOSIS; COMPONENT-C3 AB The malaria parasite Plasmodium falciparum induces a number of novel adhesion properties in the erythrocytes that it infects. One of these properties, the ability of infected erythrocytes to bind uninfected erythrocytes to form rosettes, is associated with severe malaria and may play a direct role in the pathogenesis of disease. Previous work has shown that erythrocytes deficient in complement receptor (CR) 1 (CR1, CD35; C3b/C4b receptor) have greatly reduced resetting capacity, indicating an essential role for CR1 in rosette formation. Using deletion mutants and mAbs, we have localized the region of CR1 required for the formation of P. falciparum rosettes to the area of long homologous repeat regions B and C that also acts as the binding site for the activated complement component C3b, This result raises the possibility that C3b could be an intermediary in resetting, bridging between the infected erythrocyte and CR1. We were able to exclude this hypothesis, however, as parasites grown in C3-deficient human serum formed rosettes normally. We have also shown in this report that rosettes can be reversed by mAb J3B11 that recognizes the C3b binding site of CR1. This rosette-reversing activity was demonstrated in a range of laboratory-adapted parasite strains and field isolates from Kenya and Malawi, Thus, we have mapped the region of CR1 required for resetting and demonstrated that the CR1 dependent resetting mechanism occurs commonly in P, falciparum isolates, and could therefore be a potential target for future therapeutic interventions to treat severe malaria. C1 Univ Edinburgh, Inst Cell Anim & Populat Biol, Edinburgh EH9 3JT, Midlothian, Scotland. Univ Malawi, Coll Med, Wellcome Trust Res Labs, Blantyre, Malawi. Med Coll Penn & Hahnemann Univ, Sch Med, Dept Microbiol & Immunol, Philadelphia, PA 19102 USA. Hop Broussais, Inst Natl Sante, Rech Med Unite 430, F-75674 Paris, France. KEMRI Ctr Geog Med Res Coast, Wellcome Trust Program, Kenya Med Res Inst, Kilifi, Kenya. Univ Oxford, Inst Mol Med, Oxford, England. Washington Univ, Sch Med, Dept Med, Div Rheumatol, St Louis, MO 63110 USA. NIAID, NIH, Bethesda, MD 20892 USA. RP Rowe, JA (reprint author), Univ Edinburgh, Inst Cell Anim & Populat Biol, W Mains Rd, Edinburgh EH9 3JT, Midlothian, Scotland. OI Rogerson, Stephen/0000-0003-4287-1982; Newbold, Chris/0000-0002-9274-3789 FU NIAID NIH HHS [AI RO1-42367] NR 38 TC 72 Z9 73 U1 0 U2 2 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD DEC 1 PY 2000 VL 165 IS 11 BP 6341 EP 6346 PG 6 WC Immunology SC Immunology GA 376PZ UT WOS:000165467100041 PM 11086071 ER PT J AU Yen, CH Yang, YC Ruscetti, SK Kirken, RA Dai, RM Li, CCH AF Yen, CH Yang, YC Ruscetti, SK Kirken, RA Dai, RM Li, CCH TI Involvement of the ubiquitin-proteasome pathway in the degradation of nontyrosine kinase-type cytokine receptors of IL-9, IL-2, and erythropoietin SO JOURNAL OF IMMUNOLOGY LA English DT Article ID CELL ANTIGEN RECEPTOR; TYROSINE PHOSPHORYLATION; GAMMA-CHAIN; ENDOPLASMIC-RETICULUM; SIGNAL-TRANSDUCTION; GROWTH-FACTOR; SACCHAROMYCES-CEREVISIAE; MAMMALIAN HOMOLOG; ALPHA-CHAIN; PROTEIN AB The ubiquitin-dependent proteasome-mediated (Ub-Pr) degradation pathway has been shown to regulate a large variety of substrates, including nuclear, cytosolic, and membrane proteins. In mammalian systems, polyubiquitin modification has been identified in a number of cell surface receptors for more than a decade; however, its biological significance has remained unclear until recently, For growth factor receptors vith intrinsic tyrosine kinase domains, polyubiquitination is believed to trigger the internalization and subsequent degradation via the lysosomal pathway, In this study we provide the first evidence that non-tyrosine kinase-type cytokine surface receptors, IL-9R alpha -chain, IL-2 receptor beta -chain, and erythropoietin receptor, can be polyubiquitinated and degraded by proteasomes. The Ub-Pr pathway regulates both the basal level turnover and the ligand-induced degradation of the receptors, A previously identified putative molecular chaperon, valosin-containing protein, undergoes tyrosine phosphorylation in a cytokine dependent manner and associates with the receptor complexes following receptor engagement, suggesting that valosin-containing protein may target the ubiquitinated receptors to the proteasome for degradation. C1 NCI, Frederick Canc Res & Dev Ctr, Intramural Res Support Program, Sci Applicat Int Corp, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, Div Basic Sci, Basic Res Lab, Frederick, MD 21702 USA. Indiana Univ, Sch Med, Dept Med & Biochem Mol Biol, Indianapolis, IN 46202 USA. Univ Texas, Dept Integrat Biol & Pharmacol, Houston, TX 77030 USA. RP Li, CCH (reprint author), NCI, Frederick Canc Res & Dev Ctr, Lab Leukocyte Biol, Sci Applicat Int Corp, POB B,Bldg 567,Room 252, Frederick, MD 21702 USA. FU NCI NIH HHS [N01CO56000] NR 47 TC 41 Z9 41 U1 0 U2 1 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD DEC 1 PY 2000 VL 165 IS 11 BP 6372 EP 6380 PG 9 WC Immunology SC Immunology GA 376PZ UT WOS:000165467100045 PM 11086075 ER PT J AU Sidney, J Dzuris, JL Newman, MJ Johnson, RP Amitinder, K Walker, CM Appella, E Mothe, B Watkins, DI Sette, A AF Sidney, J Dzuris, JL Newman, MJ Johnson, RP Amitinder, K Walker, CM Appella, E Mothe, B Watkins, DI Sette, A TI Definition of the Mamu A*01 peptide binding specificity: Application to the identification of wild-type and optimized ligands from simian immunodeficiency virus regulatory proteins SO JOURNAL OF IMMUNOLOGY LA English DT Article ID SECONDARY ANCHOR RESIDUES; T-LYMPHOCYTE RESPONSES; CLASS-I MOLECULES; CTL EPITOPES; HIV-1; VACCINE; COMPLEX; MOTIFS; NEF; IMMUNOGENICITY AB Single amino acid substitution analogs of the known Mamu A*01 binding peptide gag 181-190 and libraries of naturally occurring sequences of viral or bacterial origin were used to rigorously define the peptide binding motif associated,vith Mamu A*01 molecules. The presence of S or T in position 2, P in position 3, and hydrophobic or aromatic residues at the C terminus is associated with optimal binding capacity, At each of these positions, additional residues are also tolerated but associated with significant decreases in binding capacity, The presence of at least two preferred and one tolerated residues at the three anchor positions is necessary for good Mamu A*01 binding;, optimal ligand size is 8-9 residues. This detailed motif has been used to map potential epitopes from SIVmac239 regulatory proteins and to engineer peptides with increased binding capacity. A total of 13 wild type and 17 analog candidate epitopes were identified. Furthermore, our analysis reveals a significantly lower than expected frequency of epitopes in early regulatory proteins, suggesting a possible evolutionary- and/or immunoselection directed against variants of viral products that contain CTL epitopes. C1 Epimmune Inc, San Diego, CA 92121 USA. New England Reg Primate Res Ctr, Southborough, MA 01772 USA. Ohio State Univ, Coll Med & Publ Hlth, Childrens Res Inst, Columbus, OH 43205 USA. NCI, NIH, Bethesda, MD 20892 USA. Univ Wisconsin, Wisconsin Reg Primate Res Ctr, Madison, WI USA. RP Sette, A (reprint author), Epimmune Inc, 5820 Nancy Ridge Dr,Suite 100, San Diego, CA 92121 USA. OI Dzuris, John/0000-0002-0046-8426 FU NIAID NIH HHS [AI38620, N01-AI-95362, AI38081-03] NR 56 TC 42 Z9 43 U1 0 U2 2 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD DEC 1 PY 2000 VL 165 IS 11 BP 6387 EP 6399 PG 13 WC Immunology SC Immunology GA 376PZ UT WOS:000165467100047 PM 11086077 ER PT J AU Belyakov, IM Ahlers, JD Clements, JD Strober, W Berzofsky, JA AF Belyakov, IM Ahlers, JD Clements, JD Strober, W Berzofsky, JA TI Interplay of cytokines and adjuvants in the regulation of mucosal and systemic HIV-specific CTL SO JOURNAL OF IMMUNOLOGY LA English DT Article ID HEAT-LABILE ENTEROTOXIN; ADP-RIBOSYLTRANSFERASE ACTIVITY; CYTOTOXIC T-LYMPHOCYTES; HUMAN-IMMUNODEFICIENCY-VIRUS; ESCHERICHIA-COLI ENTEROTOXIN; CHOLERA-TOXIN; CLASS-I; A-SUBUNIT; NONTOXIC MUTANT; TH2 CELLS AB We examined the interplay between cytokines and adjuvants to optimize the induction of CTL by a mucosal HIV peptide vaccine. We show synergy between IL-12 and GM-CSF when administered together with the HIV peptide PCLUS3-18IIIB and cholera toxin (CT) in the induction of CTL activity and protection against mucosal viral transmission, Further, we examine the efficacy of mutant Escherichia coli labile toxin, LT(R192G), as a less toxic adjuvant than CT. LT(R192G) was as effective as or more effective than CT at inducing a mucosal CTL response. Moreover, LT(R192G) was as effective without IL-12 as CT was when combined with IL-12, and the response elicited by LT(R192G) with the vaccine was not further enhanced by the addition of IL-12. GM-CSF synergized,vith LT(R192G) without exogenous IL-12, Therefore, LT(R192G) may induce a more favorable cytokine response by not inhibiting IL-12 production, In particular, less IL-4 is made after LT(R192G) than CT immunization, and the response is less susceptible to anti-IL-12 inhibition. Thus, the choice of mucosal adjuvant affects the cytokine environment, and the mucosal response and protection can be enhanced by manipulating the cytokine environment with synergistic cytokine combinations incorporated in the vaccine. C1 NCI, Mol Immunogenet & Vaccine Res Sect, Metab Branch, NIH, Bethesda, MD 20892 USA. Tulane Univ, Sch Med, Dept Microbiol & Immunol, New Orleans, LA 70112 USA. NIAID, Mucosal Immun Sect, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. RP Berzofsky, JA (reprint author), NCI, Mol Immunogenet & Vaccine Res Sect, Metab Branch, NIH, Bldg 10,Room 6B-12,MSC 1578, Bethesda, MD 20892 USA. NR 71 TC 71 Z9 74 U1 0 U2 1 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD DEC 1 PY 2000 VL 165 IS 11 BP 6454 EP 6462 PG 9 WC Immunology SC Immunology GA 376PZ UT WOS:000165467100055 PM 11086085 ER PT J AU Nanki, T Hayashida, K El-Gabalawy, HS Suson, S Shi, KR Girschick, HJ Yavuz, S Lipsky, PE AF Nanki, T Hayashida, K El-Gabalawy, HS Suson, S Shi, KR Girschick, HJ Yavuz, S Lipsky, PE TI Stromal cell-derived factor-1-CXC chemokine receptor 4 interactions play a central role in CD4(+) T cell accumulation in rheumatoid arthritis synovium SO JOURNAL OF IMMUNOLOGY LA English DT Article ID HUMAN INTERLEUKIN-8 RECEPTOR; PROTEIN-COUPLED RECEPTOR; MOLECULAR-CLONING; FUNCTIONAL EXPRESSION; PERIPHERAL-BLOOD; SURFACE EXPRESSION; PEPTIDE RECEPTORS; EOTAXIN RECEPTOR; SINGLE-CELL; LYMPHOCYTES AB Rheumatoid arthritis (RA) is characterized by the accumulation of CD4(+) memory T cells in the inflamed synovium, To address the mechanism, we analyzed chemokine receptor expression and found that the frequency of CXC chemokine receptor (CXCR)4 expressing synovial tissue CD4(+) memory T cells was significantly elevated. CXCR4 expression could be enhanced by IL-15, whereas stromal cell-derived factor (SDF)-1, the ligand of CXCR4, was expressed in the RA synovium and could be increased by CD40 stimulation, SDF-1 stimulated migration of rheumatoid synovial T cells and also inhibited activation-induced apoptosis of T cells. These results indicate that SDP-1-CXCR4 interactions play important roles in CD4(+) memory T cell accumulation in the RA synovium, and emphasize the role of stromal cells in regulating rheumatoid inflammation. C1 NIAMSD, Bethesda, MD 20892 USA. Univ Texas, SW Med Ctr, Dept Internal Med, Dallas, TX 75235 USA. Univ Texas, SW Med Ctr, Harold C Simmons Arthrit Res Ctr, Dallas, TX 75235 USA. Osaka Univ, Sch Med, Dept Orthoped Surg, Osaka, Japan. RP Lipsky, PE (reprint author), NIAMSD, Bldg 10,Room 9N228,10 Ctr Dr,MSC 1820, Bethesda, MD 20892 USA. FU NIAMS NIH HHS [AR-36169] NR 69 TC 312 Z9 332 U1 0 U2 6 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD DEC 1 PY 2000 VL 165 IS 11 BP 6590 EP 6598 PG 9 WC Immunology SC Immunology GA 376PZ UT WOS:000165467100073 PM 11086103 ER PT J AU Zhu, JH Quyyumi, AA Norman, JE Costello, R Csako, G Epstein, SE AF Zhu, JH Quyyumi, AA Norman, JE Costello, R Csako, G Epstein, SE TI The possible role of hepatitis A virus in the pathogenesis of atherosclerosis SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID C-REACTIVE PROTEIN; CORONARY-ARTERY DISEASE; ACUTE MYOCARDIAL-INFARCTION; HEART-DISEASE; RISK; CYTOMEGALOVIRUS; INFECTION; INFLAMMATION; ASSOCIATION; CHLAMYDIA AB The possible association between hepatitis A virus (HAV) infection and coronary artery disease (CAD) was studied. Blood from 391 patients undergoing coronary angiography was tested for serum IgG antibodies to HAV and C-reactive protein (CRP), Of the 391 patients, 205 (52%) had anti-HAV IgG antibodies. CAD prevalence was 74% in HAV-seropositive and 52% in HAV-seronegative patients (P < .0001); significance persisted after adjustment for either traditional CAD risk factors or for risk factors plus other infectious agents (cytomegalovirus, Chlamydia pneumoniae, Helicobacter pylori, and herpes simplex virus). In addition, CRP levels were significantly higher in HAV-seropositive than in HAV-seronegative patients (P = .013) in both univariate and multivariate analyses. Logistic regression analysis demonstrated that HAV seropositivity is an independent predictor of risk for CAD and elevated CRP levels. HAV infection is therefore associated with CAD, which raises the possibility that this virus may play a causal role in atherogenesis. C1 Washington Hosp Ctr, CRI, MedStar Res Inst, Washington, DC 20010 USA. NHLBI, Cardiol Branch, Bethesda, MD 20892 USA. NHLBI, Div Epidemiol & Clin Applicat, Bethesda, MD 20892 USA. NIH, Ctr Clin, Bethesda, MD 20892 USA. RP Epstein, SE (reprint author), Washington Hosp Ctr, CRI, MedStar Res Inst, 110 Irving St NW,Ste 4B-1, Washington, DC 20010 USA. NR 15 TC 64 Z9 67 U1 1 U2 2 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD DEC PY 2000 VL 182 IS 6 BP 1583 EP 1587 DI 10.1086/317613 PG 5 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 378QY UT WOS:000165596100001 PM 11069227 ER PT J AU Frenkel, LM Capparelli, EV Dankner, WM Xu, J Smith, IL Ballow, A Culnane, M Read, JS Thompson, M Mohan, KM Shaver, A Robinson, CA Stempien, MJ Burchett, SK Melvin, AJ Borkowsky, W Petru, A Kovacs, A Yogev, R Goldsmith, J McFarland, EJ Spector, SA AF Frenkel, LM Capparelli, EV Dankner, WM Xu, J Smith, IL Ballow, A Culnane, M Read, JS Thompson, M Mohan, KM Shaver, A Robinson, CA Stempien, MJ Burchett, SK Melvin, AJ Borkowsky, W Petru, A Kovacs, A Yogev, R Goldsmith, J McFarland, EJ Spector, SA CA Pediat AIDS Clin Trials Grp TI Oral ganciclovir in children: Pharmacokinetics, safety, tolerance, and antiviral effects SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID ACTIVE ANTIRETROVIRAL THERAPY; CYTOMEGALOVIRUS DISEASE; TRANSPLANT RECIPIENTS; AIDS PATIENTS; CMV RETINITIS; PREVENTION; RESISTANCE; PLASMA; INFECTION; RISK AB The pharmacokinetics, safety, tolerance, and antiviral effects of ganciclovir (Gcv) administered orally were evaluated in 36 children infected with cytomegalovirus (CMV) who were severely immunocompromised by infection with human immunodeficiency virus type 1, In this dose-escalation study, 30 mg/kg of Gcv administered every 8 h produced serum levels similar to the dose (1 g/8 h) effective for maintenance treatment of CMV retinitis in adults. In older children, serum Gcv concentrations were similar after the administration of capsules and suspension. All doses (10-50 mg/kg/8 h) studied were safe and, except for the volume of suspension or number of pills, were well tolerated. Oral Gcv was associated with a decrease in the detection of CMV by culture or polymerase chain reaction, CMV disease occurred in 3 children during the study: one developed Gcv resistance, another had harbored resistant virus at study entry, and a third had wild-type CMV. C1 Univ Washington, Dept Pediat & Lab Med, Div Infect Dis, Seattle, WA 98105 USA. Univ Washington, Dept Pediat, Seattle, WA 98105 USA. Univ Washington, Dept Lab Med, Seattle, WA 98105 USA. Univ Calif San Diego, Dept Pediat, San Diego, CA 92103 USA. Univ Calif San Diego, Dept Pharm, San Diego, CA 92103 USA. Childrens Hosp, Dept Infect Dis, Oakland, CA 94609 USA. Univ So Calif, Dept Pediat, Los Angeles, CA 90089 USA. Childrens Hosp Los Angeles, Dept Hematol & Oncol, Los Angeles, CA 90027 USA. Roche Global Dev, Palo Alto, CA USA. Harvard Univ, Dept Publ Hlth, Cambridge, MA 02138 USA. Harvard Univ, Dept Pediat, Cambridge, MA 02138 USA. Frontier Sci, Amherst, MA USA. NYU, Dept Pediat, New York, NY 10016 USA. NIAID, Bethesda, MD 20892 USA. NICHHD, Bethesda, MD 20892 USA. Social & Sci Syst, Rockville, MD USA. Northwestern Univ, Dept Pediat, Chicago, IL 60611 USA. Univ Colorado Hlth Sci, Dept Pediat, Denver, CO USA. RP Frenkel, LM (reprint author), Univ Washington, Dept Pediat & Lab Med, Div Infect Dis, 4800 Sand Point Way NE, Seattle, WA 98105 USA. NR 24 TC 17 Z9 17 U1 1 U2 1 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD DEC PY 2000 VL 182 IS 6 BP 1616 EP 1624 DI 10.1086/317600 PG 9 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 378QY UT WOS:000165596100006 PM 11069232 ER PT J AU Blankson, JN Finzi, D Pierson, TC Sabundayo, BP Chadwick, K Margolick, JB Quinn, TC Siliciano, RF AF Blankson, JN Finzi, D Pierson, TC Sabundayo, BP Chadwick, K Margolick, JB Quinn, TC Siliciano, RF TI Biphasic decay of latently infected CD4(+) T cells in acute human immunodeficiency virus type 1 infection SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 7th Conference on Retroviruses and Opportunistic Infections CY JAN 29-FEB 03, 2000 CL SAN FRANCISCO, CALIFORNIA ID ACTIVE ANTIRETROVIRAL THERAPY; PRIMARY HIV-1 INFECTION; COMBINATION THERAPY; IN-VIVO; LYMPHOCYTES; RESERVOIR; PLASMA; ESTABLISHMENT; PERSISTENCE; VIREMIA AB Latent infection of resting CD4(+) T cells represents a major barrier to eradication of human immunodeficiency virus type 1 (HIV-1), The establishment and rate of decay of latent HIV-1 in resting CD+ T cells from 9 acute seroconverters, 7 of whom began to receive highly active antiretroviral therapy (HAART) shortly after presentation, were studied, Before the initiation of therapy, these patients had very high frequencies of latently infected CD4(+) T cells, with a median frequency of 205 infectious units per million resting CD4(+) T cells, These values are greater than or equal to1 log higher than those seen in chronically infected patients who are not undergoing HAART. The number of latently infected cells declined dramatically after initiation of HAART but then tended to level off at a low but stable level, The biphasic decay of latent HIV in resting CD4(+) T cells in acute seroconverters supports current models of pre- and postintegration latency. C1 Johns Hopkins Univ, Sch Med, Dept Med, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Hyg & Publ Hlth, Dept Mol Microbiol & Immunol, Baltimore, MD USA. NIAID, NIH, Bethesda, MD 20892 USA. RP Siliciano, RF (reprint author), Johns Hopkins Univ, Sch Med, Dept Med, 1071S Ross Bldg,720 Rutland Ave, Baltimore, MD 21205 USA. FU NIAID NIH HHS [AI43222, AI10140, AI41532] NR 25 TC 86 Z9 89 U1 0 U2 3 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD DEC PY 2000 VL 182 IS 6 BP 1636 EP 1642 DI 10.1086/317615 PG 7 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 378QY UT WOS:000165596100008 PM 11069234 ER PT J AU Spector, SA Hsia, K Yong, FH Cabral, S Fenton, T Fletcher, CV McNamara, J Mofenson, LM Starr, SE AF Spector, SA Hsia, K Yong, FH Cabral, S Fenton, T Fletcher, CV McNamara, J Mofenson, LM Starr, SE TI Patterns of plasma human immunodeficiency virus type 1 RNA response to highly active antiretroviral therapy in infected children SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID COMBINATION THERAPY; INFANTS; DYNAMICS AB This study examined the rate of decline in plasma human immunodeficiency virus type 1 (HIV-1) RNA levels to <400 and <50 copies/ml in children receiving highly active antiretroviral therapy (HAART) consisting of efavirenz, nelfinavir, and 1 or 2 nucleoside reverse-transcriptase inhibitors. Children receiving HAART achieved a plasma HIV-1 RNA level <400 copies/ mi, by a median of 4 weeks after initiation of therapy and a decline to <50 copies/ml by 20 weeks. Baseline plasma HIV-1 RNA levels affected the likelihood of achieving potent and sustained virus suppression, and children whose CD4 lymphocyte counts increased >70 cells/ muL by 20 weeks on therapy were more likely to achieve durable virological and immunological benefit. These data provide time frames for virus suppression after the initiation of HAART that should be useful in evaluating the potential efficacy and durability of response of newly instituted combination antiretroviral therapy in HIV-1-infected children. C1 Univ Calif San Diego, Dept Pediat, Div Pediat Infect Dis, La Jolla, CA 92093 USA. Harvard Univ, Sch Publ Hlth, Dept Biostat, Boston, MA 02115 USA. Univ Minnesota, Dept Expt & Clin Pharmacol, Minneapolis, MN USA. NIAID, Rockville, MD 20852 USA. NICHD, Rockville, MD USA. Childrens Hosp Philadelphia, Div Immunol & Infect Dis, Philadelphia, PA 19104 USA. RP Spector, SA (reprint author), Univ Calif San Diego, Dept Pediat, Div Pediat Infect Dis, 9500 Gilman Dr, La Jolla, CA 92093 USA. OI Mofenson, Lynne/0000-0002-2818-9808 FU NIAID NIH HHS [AI-27563, AI-39004, AI-41110] NR 13 TC 24 Z9 25 U1 0 U2 1 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD DEC PY 2000 VL 182 IS 6 BP 1769 EP 1773 DI 10.1086/317621 PG 5 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 378QY UT WOS:000165596100026 PM 11069252 ER PT J AU Shearer, WT Israel, RJ Starr, S Fletcher, CV Wara, D Rathore, M Church, J DeVille, J Fenton, T Graham, B Samson, P Staprans, S McNamara, J Moye, J Maddon, PJ Olson, WC AF Shearer, WT Israel, RJ Starr, S Fletcher, CV Wara, D Rathore, M Church, J DeVille, J Fenton, T Graham, B Samson, P Staprans, S McNamara, J Moye, J Maddon, PJ Olson, WC CA Pediat AIDS Clin Trials Grp Proto TI Recombinant CD4-IgG2 in human immunodeficiency virus type 1-infected children: Phase 1/2 study SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 7th Conference on Retroviruses and Opportunistic Infections CY JAN 29-FEB 03, 2000 CL SAN FRANCISCO, CALIFORNIA ID AIDS-RELATED COMPLEX; SOLUBLE CD4; PHARMACOKINETICS; NEUTRALIZATION; SAFETY; CD4-IMMUNOGLOBULIN-G; ANTIBODIES; TRIAL AB The use of recombinant CD4-IgG2 in pediatric human immunodeficiency virus type 1 (HIV-1) infection was evaluated by single and multidose intravenous infusions in 18 children in a phase 1/2 study. The study drug was well tolerated, and dose proportionality was observed in terms of area under time-concentration curve and peak serum concentration. Acute decreases of >0.7 log(10) copies/mL in serum HIV-1 RNA concentration were seen in 4 of the 6 children treated with 4 weekly 10 mg/kg doses. At 14 days after treatment, 3 children had sustained reductions in serum HIV-I RNA; the other children had rebounded to baseline levels or above. By 28 days after therapy, the peak HIV-1 cellular infectious units was reduced in all 6 children, including the 2 who had experienced an earlier transient increase in values. Thus, recombinant CD4-IgG2 treatment of HIV-l-infected children appears to be well tolerated and capable of reducing HIV-1 burden. C1 Baylor Coll Med, Dept Pediat, Houston, TX 77030 USA. Baylor Coll Med, Dept Immunol, Houston, TX 77030 USA. Progen Pharmaceut Inc, Tarrytown, NY USA. Childrens Hosp Philadelphia, Div Infect Dis & Immunol, Philadelphia, PA 19104 USA. Univ Minnesota, Div Clin Pharmacol, Minneapolis, MN USA. Univ Calif San Francisco, Dept Pediat, San Francisco, CA 94143 USA. Childrens Hosp Los Angeles, Childrens AIDS Ctr, Los Angeles, CA 90027 USA. Univ Calif Los Angeles, Dept Pediat, Los Angeles, CA 90024 USA. Univ Florida, Hlth Sci Ctr Jacksonville, Div Infect Dis Immunol, Jacksonville, FL 32209 USA. Frontier Sci & Technol Res Fdn Inc, Stat & Data Anal Ctr, Brookline, MA USA. Stat & data Management Ctr, Amherst, NY USA. Harvard Univ, Sch Publ Hlth, Stat & Data Anal Ctr, Boston, MA 02115 USA. Emory Univ, Sch Med, Dept Microbiol & Immunol, Atlanta, GA 30322 USA. NIAID, Pediat Med Branch, Div Aids, Bethesda, MD 20892 USA. NICHHD, Pediat Adolescent & Maternal AIDS Branch, Ctr Res Mothers & Children, Bethesda, MD 20892 USA. RP Shearer, WT (reprint author), Baylor Coll Med, Dept Pediat Allergy Immunol, 6621 Fannin St,MC 1-3291, Houston, TX 77030 USA. OI moye, john/0000-0001-9976-8586 NR 16 TC 58 Z9 62 U1 0 U2 1 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD DEC PY 2000 VL 182 IS 6 BP 1774 EP 1779 DI 10.1086/317622 PG 6 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 378QY UT WOS:000165596100027 PM 11069253 ER PT J AU Weil, GJ Steel, C Liftis, F Li, BW Mearns, G Lobos, E Nutman, TB AF Weil, GJ Steel, C Liftis, F Li, BW Mearns, G Lobos, E Nutman, TB TI A rapid-format antibody card test for diagnosis of onchocerciasis SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID ANTIGENS; INFECTION AB Improved methods are needed for field diagnosis of onchocerciasis, to support efforts aimed at elimination of the disease. A rapid-format card test was evaluated that detects IgG4 antibodies to recombinant Onchocerca volvulus antigen Ov16 with serum samples from patients with onchocerciasis and with various types of control serum samples. The sensitivity of the test with serum samples from 106 microfilariae-positive subjects was 90.6%. The test was equally sensitive with serum samples obtained from patients in Africa and Latin America. Specificity was excellent; positive tests were observed for 2 of 38 serum samples from patients with other filarial infections and for 1 of 23 serum samples from patients with nonfilarial helminth infections. The 3 "falsepositive" serum samples were from West Africans who could have been coinfected with onchocerciasis. No positive tests were observed with nonendemic serum samples from normal adults, patients with autoimmune disorders, or patients with the hyper-IgE syndrome. This new test holds great promise as a simple tool for diagnosis of onchocerciasis. C1 Barnes Jewish Hosp, Div Infect Dis, St Louis, MO 63110 USA. Washington Univ, Sch Med, St Louis, MO USA. NIH, Parasit Dis Lab, Bethesda, MD 20892 USA. AMRAD ICT, Sydney, NSW, Australia. UCL, London, England. RP Weil, GJ (reprint author), Barnes Jewish Hosp, Div Infect Dis, 216 S Kingshighway, St Louis, MO 63110 USA. EM gweil@im.wustl.edu FU NIAID NIH HHS [AI-22488] NR 15 TC 55 Z9 55 U1 0 U2 5 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 1537-6613 J9 J INFECT DIS JI J. Infect. Dis. PD DEC PY 2000 VL 182 IS 6 BP 1796 EP 1799 DI 10.1086/317629 PG 4 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 378QY UT WOS:000165596100032 PM 11069258 ER PT J AU Gopinath, R Ostrowski, M Justement, SJ Fauci, AS Nutman, TB AF Gopinath, R Ostrowski, M Justement, SJ Fauci, AS Nutman, TB TI Filarial infections increase susceptibility to human immunodeficiency virus infection in peripheral blood mononuclear cells in vitro SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 48th Annual Meeting of the American-Society-of-Tropical-Medicine-and-Hygiene CY NOV 28-DEC 02, 1999 CL WASHINGTON, D.C. SP Amer Soc Trop Med & Hyg ID DISEASE PROGRESSION; TYPE-1 REPLICATION; CCR5 EXPRESSION; TRANSMISSION; ACTIVATION; CYTOKINES; IL-10 AB Because helminth infections and human immunodeficiency virus (HIV) coexist in areas where the spread of AIDS is most dramatic, their in vitro interaction was explored. Cryopreserved peripheral blood mononuclear cells (PBMC) from patients with filarial infections (n = 24) and from unexposed control subjects (n = 12) were depleted of CD8 T cells and were infected with macrophage (M)- and T cell-tropic viruses. A trend toward increased HIV replication in PBMC from filaria-infected patients was observed. Furthermore, PBMC from 6 filaria-infected patients before antifilarial treatment were significantly more susceptible to replication of M-tropic virus than their posttreatment PBMC (P = .03), No intergroup differences were found in the surface expression of HLA-DR, CD25, CCR5, CXCR4, CCR3 on CD4 T cells, or monocytes before infection, PBMC from filaria-infected patients produced less RANTES (P = .02) but more intracellular interleukin-4 than those of control subjects. Thus, PBMC from persons with filarial infections appear to have enhanced susceptibility to HIV-1 infection mediated by an undetermined mechanism. C1 NIAID, Parasit Dis Lab, Helminth Immunol Sect, NIH, Bethesda, MD 20892 USA. NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. RP Gopinath, R (reprint author), NIAID, Parasit Dis Lab, Helminth Immunol Sect, NIH, 4 Ctr Dr,Room 126, Bethesda, MD 20892 USA. NR 15 TC 62 Z9 64 U1 0 U2 2 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD DEC PY 2000 VL 182 IS 6 BP 1804 EP 1808 DI 10.1086/317623 PG 5 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 378QY UT WOS:000165596100034 PM 11069260 ER PT J AU Slor, H Batko, S Khan, SG Sobe, T Emmert, S Khadavi, A Frumkin, A Busch, DB Albert, RB Kraemer, KH AF Slor, H Batko, S Khan, SG Sobe, T Emmert, S Khadavi, A Frumkin, A Busch, DB Albert, RB Kraemer, KH TI Clinical, cellular, and molecular features of an Israeli xeroderma pigmentosum family with a frameshift mutation in the XPC gene: Sun protection prolongs life SO JOURNAL OF INVESTIGATIVE DERMATOLOGY LA English DT Article DE cancer genetics; DNA polymorphism; DNA repair; skin cancer; sun protection ID NUCLEOTIDE EXCISION-REPAIR; GROUP-C PROTEIN; DNA-REPAIR; CELLS; EXPRESSION; CLONING; COMPLEX AB An Ashkenazi Jewish Israeli family with two children affected with severe xeroderma pigmentosum was investigated, A son, XP12TA, developed skin cancer at 2 y and died at 10 y. A daughter, XP25TA, now 24 y old, was sun protected and began developing skin cancers at 10 y. Their cultured skin fibroblasts showed reductions in post-ultraviolet survival (11% of normal), unscheduled DNA synthesis (10% of normal), global genome DNA repair (15% of normal), and plasmid host cell reactivation (5% of normal). Transcription-coupled DNA repair was normal, however, Northern blot analysis revealed greatly reduced xeroderma pigmentosum complementation group C mRNA, A plasmid host cell reactivation assay assigned the cells to xeroderma pigmentosum complementation group C, Cells from both parents and an unaffected child exhibited normal post-ultraviolet-C survival and normal DNA repair. Sequencing the xeroderma pigmentosum complementation group C cDNA of XP12TA and XP25TA revealed a homozygous deletion of two bases (del AT 669-670) in exon 5 with a new termination site 10 codons downstream that is expected to encode a truncated xeroderma pigmentosum complementation group C protein. Sequence analysis of the xeroderma pigmentosum complementation group C cDNA in cells from the parents found identical heterozygous mutations: one allele carries both the exon 5 frameshift and an exon 15 polymorphism and the other allele carries neither alteration. Cells from the unaffected brother had two normal xeroderma pigmentosum complementation group C alleles, This frameshift mutation in the xeroderma pigmentosum complementation group C gene led to reduced DNA repair with multiple skin cancers and early death. Sun protection delayed the onset of skin cancer and prolonged Life in a sibling with the same mutation. C1 NCI, Basic Res Lab, Bethesda, MD 20892 USA. Tel Aviv Univ, Sackler Fac Med, Dept Human Genet & Mol Med, Tel Aviv, Israel. Kaplan Hosp, Dept Dermatol, Rehovot, Israel. Armed Forces Inst Pathol, Dept Environm & Toxicol Pathol, Washington, DC USA. RP Kraemer, KH (reprint author), NCI, Basic Res Lab, Bldg 37 Room 3E24, Bethesda, MD 20892 USA. FU Intramural NIH HHS [Z01 BC004517-31] NR 27 TC 21 Z9 21 U1 0 U2 0 PU BLACKWELL SCIENCE INC PI MALDEN PA 350 MAIN ST, MALDEN, MA 02148 USA SN 0022-202X J9 J INVEST DERMATOL JI J. Invest. Dermatol. PD DEC PY 2000 VL 115 IS 6 BP 974 EP 980 DI 10.1046/j.1523-1747.2000.00190.x PG 7 WC Dermatology SC Dermatology GA 390FT UT WOS:000166285600007 PM 11121128 ER PT J AU Chen, CL Yull, FE Cardwell, N Singh, N Strayhorn, WD Nanney, LB Kerr, LD AF Chen, CL Yull, FE Cardwell, N Singh, N Strayhorn, WD Nanney, LB Kerr, LD TI RAG2-/-, I kappa B-alpha-/- chimeras display a psoriasiform skin disease SO JOURNAL OF INVESTIGATIVE DERMATOLOGY LA English DT Article DE I kappa B-alpha; keratinocytes; lymphocytes; psoriasis ID TRANSGENIC MICE; NUCLEAR-FACTOR; IMMUNE-RESPONSES; T-CELLS; PSORIATIC PLAQUES; CYTOKINE NETWORK; ANIMAL-MODELS; NUDE-MICE; ACTIVATION; EXPRESSION AB Nuclear factor-kappaB, a ubiquitous transcription factor involved in inflammatory and immune responses, is inappropriately activated in several immune-related diseases, such as allograft rejection, or bronchial asthma. As nuclear factor-kappaB activity is regulated by inhibitor of kappaB (I kappaB), the gene encoding I kappaB-alpha was disrupted in mice to observe the in vivo effects of hyperactivation of nuclear factor-kappaB. I kappaB-alpha-/- mice have constitutive nuclear factor-kappaB activity, severe skin disease, and neonatal lethality. To determine the role of I kappaB-alpha deficient immunocytes in the pathogenesis of the skin disease in adult mice, we utilized the RAG2-deficient blastocyst complementation system to generate RAG2-/-, I kappaB-alpha-/- chimeras. These animals display a psoriasiform dermatitis characterized by hyperplastic epidermal keratinocytes and dermal infiltration of immunocytes, including lymphocytes. Skin grafts transferred from diseased chimeras to recipient nude mice produce hyperproliferative psoriasiform epidermal keratinocytes in response to stimulation. Furthermore, adoptive transfer of lymph node cells from diseased chimeras to RAG2-/- recipient mice recapitulates the disease. Taken together, these characterizations provide evidence to suggest that constitutive activation of nuclear factor-kappaB, due to deficiency in I kappaB-alpha, can invoke severe psoriasiform dermatitis in adult mice. C1 Vanderbilt Univ, Sch Med, Vanderbilt Ingram Canc Ctr, Dept Microbiol & Immunol, Nashville, TN 37232 USA. Vanderbilt Univ, Sch Med, Vanderbilt Ingram Canc Ctr, Dept Cell Biol, Nashville, TN 37232 USA. Vanderbilt Univ, Sch Med, Vanderbilt Ingram Canc Ctr, Dept Plast Surg, Nashville, TN 37232 USA. NIH, Transplantat & Immunogenet Sect, Div Allergy Immunol & Infect Dis, Bethesda, MD USA. RP Yull, FE (reprint author), Vanderbilt Univ, Sch Med, Vanderbilt Ingram Canc Ctr, Dept Microbiol & Immunol, A-4320 MCN,1161 21st Ave S, Nashville, TN 37232 USA. FU NCI NIH HHS [CA 68485]; NIAMS NIH HHS [P30AR41943]; NIGMS NIH HHS [R01 GM51249] NR 60 TC 15 Z9 16 U1 0 U2 0 PU BLACKWELL SCIENCE INC PI MALDEN PA 350 MAIN ST, MALDEN, MA 02148 USA SN 0022-202X J9 J INVEST DERMATOL JI J. Invest. Dermatol. PD DEC PY 2000 VL 115 IS 6 BP 1124 EP 1133 DI 10.1046/j.1523-1747.2000.00162.x PG 10 WC Dermatology SC Dermatology GA 390FT UT WOS:000166285600030 PM 11121151 ER PT J AU Frumento, G Ottonello, L Bertolotto, M Franchello, S Melioli, G Dallegri, F AF Frumento, G Ottonello, L Bertolotto, M Franchello, S Melioli, G Dallegri, F TI Spontaneous apoptosis in neutrophils is associated with downregulation of HLA Class I and is prevented by ligation of Class I SO JOURNAL OF LEUKOCYTE BIOLOGY LA English DT Article DE human; neutrophils; age-related; apoptosis ID MAJOR HISTOCOMPATIBILITY COMPLEX; MHC CLASS-I; PROGRAMMED CELL-DEATH; HUMAN POLYMORPHONUCLEAR LEUKOCYTES; GROWTH FACTOR-I; FC-GAMMA-RIII; T-CELLS; MACROPHAGE PHAGOCYTOSIS; SIGNAL-TRANSDUCTION; AGING NEUTROPHILS AB In many types of cells, ligation of human leukocyte antigens (HLA) Class I molecules with specific mAbs results in the transduction of signals that trigger different cell functions. We have investigated the effects of Class I ligation in human neutrophils, After several hours in culture, neutrophils split spontaneously into two subpopulations, one with normal and the other with reduced levels of Class I. The latter subpopulation displayed high binding capacity for Annexin V, showed a hypodiploid peak, electrophoretic DNA fragmentation, and morphological features of apoptotic cells. The addition of drugs known to delay apoptosis (GM-CSF or cAMP) resulted in a reduction of Class I modulation. Furthermore, ligation of surface Class I with F(ab')(2) fragments of the anti-Class I mAb W6/32 resulted in a delay in the progression of apoptosis, These data indicate that this surface Class I molecule is a marker of age-related apoptosis, and the ligation of these molecules results in the transduction of a signal that inhibits apoptosis, Thus, the downregulation of HLA Class I molecules in aging neutrophils prevents their halting the apoptotic process. C1 Natl Inst Canc Res, Immunogenet Lab, Genoa, Italy. Natl Inst Canc Res, Flow Cytometry Unit, Genoa, Italy. Univ Genoa, Dept Internal Med, I-16126 Genoa, Italy. RP Frumento, G (reprint author), Natl Canc Inst, Adv Biotechnol Ctr A2, Immunogenet Lab, Largo Rosanna Benzi 10, I-16132 Genoa, Italy. OI DALLEGRI, FRANCO/0000-0001-9537-4547; Bertolotto, Maria Bianca/0000-0002-4889-8046 NR 48 TC 9 Z9 9 U1 1 U2 3 PU FEDERATION AMER SOC EXP BIOL PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0741-5400 J9 J LEUKOCYTE BIOL JI J. Leukoc. Biol. PD DEC PY 2000 VL 68 IS 6 BP 873 EP 880 PG 8 WC Cell Biology; Hematology; Immunology SC Cell Biology; Hematology; Immunology GA 381FE UT WOS:000165751000012 PM 11129655 ER PT J AU Pratley, RE Baier, L Pan, DA Salbe, AD Storlien, L Ravussin, E Bogardus, C AF Pratley, RE Baier, L Pan, DA Salbe, AD Storlien, L Ravussin, E Bogardus, C TI Effects of an Ala54Thr polymorphism in the intestinal fatty acid-binding protein on responses to dietary fat in humans SO JOURNAL OF LIPID RESEARCH LA English DT Article DE nonesterified fatty acids; triglycerides; insulin; insulin resistance; genetic studies ID INSULIN-RESISTANCE; DIABETES-MELLITUS; PIMA-INDIANS; CODON-54 POLYMORPHISM; OBESE SUBJECTS; GENE; NIDDM; SUBSTITUTION; PATHOGENESIS; SENSITIVITY AB A polymorphism in FABP2 that results in an alanine-to-threonine substitution at amino acid 54 of the intestinal fatty acid-binding protein (IFABP) is associated with insulin resistance in Pima Indians. In vitro, the threonine form (Thr54) has a higher binding affinity for long-chain fatty acids than does the alanine form (Ala54). We tested whether this polymorphism affected metabolic responses to dietary fat, in vivo. Eighteen healthy Pima Indians, half homozygous for the Thr54 form of IFABP and half homozygous for the Ala54 form, were studied. The groups were matched for sex, age, and body mass index. Plasma triglyceride, nonesterified fatty acid (NEFA), glucose, and insulin responses were measured after a mixed meal (35% of daily energy requirements, 50 g of fat) and after a high fat challenge (1362 kcal, 129 g of fat). NEFA concentrations were similar to 15% higher after the mixed meal and peaked earlier and were similar to 20% higher at 7 h in response to the high fat test meal in Thr54 homozygotes compared with Ala54 homozygotes. Insulin responses to the test meals tended to be higher in Thr54 homozygotes, but glucose and triglyceride responses were not different. The results of this study suggest that the Thr54 form of IFABP is associated with higher and prolonged NEFA responses to dietary fat in vivo. Higher NEFA concentrations may contribute to insulin resistance and hyperinsulinemia in individuals with this allele. C1 NIDDKD, Clin Diabet & Nutr Sect, NIH, Phoenix, AZ 85016 USA. NIDDKD, Phoenix Epidemiol & Clin Res Branch, NIH, Phoenix, AZ 85016 USA. Univ Dundee, Dept Biochem, Dundee DD1 4HN, Scotland. Univ Wollongong, Dept Biomed Sci, Wollongong, NSW 2522, Australia. Royal Prince Alfred Hosp, Dept Endocrinol, Camperdown, NSW 2050, Australia. RP Pratley, RE (reprint author), NIDDKD, Clin Diabet & Nutr Sect, NIH, 4212 N 16Th St, Phoenix, AZ 85016 USA. NR 24 TC 50 Z9 56 U1 1 U2 3 PU LIPID RESEARCH INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0022-2275 J9 J LIPID RES JI J. Lipid Res. PD DEC PY 2000 VL 41 IS 12 BP 2002 EP 2008 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 385UG UT WOS:000166021700011 PM 11108733 ER PT J AU Uchida, Y Hara, M Nishio, H Sidransky, E Inoue, S Otsuka, F Suzuki, A Elias, PM Holleran, WM Hamanaka, S AF Uchida, Y Hara, M Nishio, H Sidransky, E Inoue, S Otsuka, F Suzuki, A Elias, PM Holleran, WM Hamanaka, S TI Epidermal sphingomyelins are precursors for selected stratum corneum ceramides SO JOURNAL OF LIPID RESEARCH LA English DT Article DE glucosylceramide; epidermis; Gaucher disease ID CULTURED HUMAN KERATINOCYTES; FETAL-RAT EPIDERMIS; PERMEABILITY BARRIER; ATOPIC-DERMATITIS; PIG EPIDERMIS; LIPID-COMPOSITION; PLASMA-MEMBRANE; MAMMALIAN EPIDERMIS; LAMELLAR GRANULES; ETIOLOGIC FACTOR AB Epidermal ceramides (Cer) comprise a heterogeneous family of seven species, including two unique omega -hydroxylated Cer, that are key components of the stratum corneum (SC) intercellular lamellar membranes responsible for the epidermal permeability barrier. Although both glucosylceramide (GlcCer) and the phospho-sphingolipid sphingomyelin (SM) are potential precursors of SC Cer, based on reported chemical structures of epidermal GlcCer and SC Cer, it is assumed that all major subfractions of SC Cer are generated from lamellar body-derived GlcCer. Yet, we and others have shown that SM-derived Cer are required for normal barrier homeostasis. Moreover, two pools of SM, one from plasma membrane, the other from lamellar body-derived contents, are potentially available for Cer production. To clarify the role of SM as a potential precursor of bulk or specific SC Cer, we compared Cer moieties in epidermal SM, Cer generated from epidermal SM by sphingomyelinase treatment, Cer within SC, and Cer that persist in Gaucher SC, where GlcCer cannot generate Cer due to an absence of beta -glucocerebrosidase. Using gas chromatography-mass spectrometry, fast atom bombardment-mass spectrometry, and nuclear magnetic resonance for Cer characterization, epidermal SM comprise three major subfractions with distinctive amide-linked (N-acyl) fatty acid (FA) compositions: that is, either long-chain FA (SM-1; C22-26), Short-chain FA (SM-2; primarily C-16), and short-chain alpha -hydroxy FA (SM-3; C16-18). In contrast, only trace quantities of omega -hydroxy FA were present. For each SM subfraction, the sphingoid base was either sphingosine or sphinganine, but phytosphingosine was not detected. Comparison of these SM with corresponding sphingomyelinase-generated epidermal Cer and SC Cer revealed that the Cer moieties of SM-1 and SM-3 are equivalent to Cer 2 (NS) and Cer 5 (AS), respectively. Moreover, both Cer 2 and Cer 5 occurred in Gaucher SC, whereas other Cer subfractions did not occur. These results indicate that two epidermal SM, that is, SM-1 and SM-3, are important precursors of two corresponding Cer in mammalian SC, that is, Cer 2 and Cer 5, but other Cer species, including the omega -hydroxy Cer species, do not derive from SM. C1 Univ Calif San Francisco, Dept Dermatol, San Francisco, CA 94143 USA. Univ Calif San Francisco, Dept Pharmaceut Chem, San Francisco, CA 94143 USA. Dept Vet Affairs Med Ctr, Dermatol Serv, San Francisco, CA 94121 USA. Dept Vet Affairs Med Ctr, Res Unit, San Francisco, CA 94121 USA. Kanebo Ltd, Cosmet Lab, Odawara, Kanagawa 2500002, Japan. NIMH, Sect Mol Neurogenet, Bethesda, MD 20892 USA. Univ Tsukuba, Dept Dermatol, Tsukuba, Ibaraki 3050006, Japan. RIKEN, Frontier Res Syst, Suprabiomol Syst Res Grp, Sphingolipid Express Lab, Wako, Saitama 3510198, Japan. RP Holleran, WM (reprint author), Univ Calif San Francisco, Dept Dermatol, San Francisco, CA 94143 USA. OI Inoue, Shintaro/0000-0002-4996-5693 FU NIAMS NIH HHS [AR39448, AR19098] NR 72 TC 121 Z9 121 U1 2 U2 7 PU LIPID RESEARCH INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0022-2275 J9 J LIPID RES JI J. Lipid Res. PD DEC PY 2000 VL 41 IS 12 BP 2071 EP 2082 PG 12 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 385UG UT WOS:000166021700019 PM 11108741 ER PT J AU Frattali, C AF Frattali, C TI On the eve of prospective payment for inpatient rehabilitation SO JOURNAL OF MEDICAL SPEECH-LANGUAGE PATHOLOGY LA English DT Article C1 NIH, Warren G Magnuson Clin Ctr, Bethesda, MD 20892 USA. RP Frattali, C (reprint author), NIH, Warren G Magnuson Clin Ctr, Bldg 10, Bethesda, MD 20892 USA. NR 7 TC 0 Z9 0 U1 0 U2 0 PU SINGULAR PUBLISHING GROUP INC PI SAN DIEGO PA 401 WEST A ST, STE 325, SAN DIEGO, CA 92101-7904 USA SN 1065-1438 J9 J MED SPEECH-LANG PA JI J. Med. Speech-Lang. Pathol. PD DEC PY 2000 VL 8 IS 4 BP XV EP XVII PG 3 WC Audiology & Speech-Language Pathology; Clinical Neurology SC Audiology & Speech-Language Pathology; Neurosciences & Neurology GA 388EU UT WOS:000166165100002 ER PT J AU Steingart, RA Solomon, B Brenneman, DE Fridkin, M Gozes, I AF Steingart, RA Solomon, B Brenneman, DE Fridkin, M Gozes, I TI VIP and peptides related to activity-dependent neurotropic factor protect PC12 cells against oxidative stress SO JOURNAL OF MOLECULAR NEUROSCIENCE LA English DT Article DE ADNP-9; H2O2; NAP; PC12; rat; RT-PCR; VIP ID VASOACTIVE-INTESTINAL-PEPTIDE; ACTING NEUROPROTECTIVE PEPTIDE; PARKINSONS-DISEASE; ALZHEIMERS-DISEASE; NEURITE OUTGROWTH; SURVIVAL; POLYPEPTIDE; ACTIVATION; NEURONS; DEATH AB Oxidative stress is a common associative mechanism that is part of the pathogenesis of many neurodegenerative diseases. Vasoactive intestinal peptide (VIP) is a principal neuropeptide associated with normal development and aging. We have previously reported that VIP induced the secretion of proteins from glial cells, including the novel survival-promoter: activity-dependent neurotrophic factor (ADNF). ADNF-9, a nine amino acid peptide derived from ADNF, protects neurons from death caused by various toxins. In the present study, we examined the neuroprotective effect of VIP against oxidative stress in a pheochromocytoma cell line (PC12). In addition, a Lipophilic derivative of VIP, Stearyl-Nle(17)-VIP (SNV), and two femtomolar-acting peptides: ADNF-9 and a 70% homologous peptide to ADNF-9, NAP were tested as well. PC12 cells were treated with 100 muM H2O2 for 24 h resulting in a reduction in cell survival to 35-50% as compared to controls. Addition of VIP or SNV prior and during the exposure to 100 muM H2O2 increased cell survival to 80-90% of control values. Culture treatment with ADNF-9 or NAP in the presence of 100 muM H2O2 increased cell survival to 75-80% of control values. Messenger RNA expression analysis revealed that incubation with VIP resulted in a twofold increase in VIP mRNA, whereas NAP treatment did not cause any change in VIP expression, implicating different mechanisms of action. Furthermore, addition of an ADNF-9 antibody prevented the ability of VIP to protect against oxidative stress, suggesting that VIP protection is partially mediated via an ADNF-like protein. C1 Tel Aviv Univ, Sackler Sch Med, Dept Clin Biochem, IL-69978 Tel Aviv, Israel. Tel Aviv Univ, George S Wise Fac Life Sci, Dept Mol Microbiol & Biotechnol, IL-69978 Tel Aviv, Israel. NICHHD, Sect Dev & Mol Pharmacol, Dev Neurobiol Lab, NIH, Bethesda, MD 20892 USA. Weizmann Inst Sci, Dept Organ Chem, IL-76100 Rehovot, Israel. RP Gozes, I (reprint author), Tel Aviv Univ, Sackler Sch Med, Dept Clin Biochem, IL-69978 Tel Aviv, Israel. NR 46 TC 67 Z9 67 U1 2 U2 5 PU HUMANA PRESS INC PI TOTOWA PA 999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USA SN 0895-8696 J9 J MOL NEUROSCI JI J. Mol. Neurosci. PD DEC PY 2000 VL 15 IS 3 BP 137 EP 145 DI 10.1385/JMN:15:3:137 PG 9 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 415XR UT WOS:000167749300001 PM 11303778 ER PT J AU Sigalov, E Fridkin, M Brenneman, DE Gozes, I AF Sigalov, E Fridkin, M Brenneman, DE Gozes, I TI VIP-related protection against iodoacetate toxicity in pheochromocytoma (PC12) cells SO JOURNAL OF MOLECULAR NEUROSCIENCE LA English DT Article DE ischemic/hypoxic injury; ischemic/hypoxic tolerance; pre-conditioning; PC12 pheochromocytoma cells; vasoactive intestinal peptide (VIP); Stearyl-Norleucine(17)-VIP (SNV); activity-dependent neuroprotective protein (ADNP); iodoacetate ID VASOACTIVE-INTESTINAL-PEPTIDE; DEPENDENT NEUROTROPHIC FACTOR; NEUROPROTECTIVE PEPTIDE; NEURONAL SURVIVAL; GLUTAMATE RELEASE; DEVELOPING BRAIN; IN-VITRO; CULTURES; HYPOXIA; GLYCOGENOLYSIS AB To evaluate the protective properties of peptides related functionally and/or structurally to vasoactive intestinal peptide (VIP), PC12 cultures were treated with iodoacetate as a model for neuronal ischemic/hypoxic injury. Brain tissue can be pre-conditioned against lethal ischemia by several mechanisms including sub-lethal ischemia, moderate hypoglycemia, heat shock, and growth factors. In the present study, a superactive VIP lipophilic analog (Stearyl-Norleucine(17)-VIP; SNV) was used to pre-condition media of PC12 cells. After removal of the conditioned media, the cultures were exposed to iodoaceate, which inhibits glycolysis. Protective efficacy against iodoacetate-induced injury was assessed by the measurements of lactate dehydrogenase (LDH) activity in the media. Treatment with iodoacetate for 2.5 h produced a twofold increase in LDH activity in the media. The protective effect of SNV had an EC50 of 1 pM. Comparison of the preconditioning time required for full protection by SNV showed no apparent difference between a 15 min and a 2 h incubation period prior to the addition of iodoacetate. Iodoacetate treatment produced a 20% decrease in the RNA transcripts encoding activity-dependent neuroprotective protein (ADNP), a novel glia-derived protein that is regulated by VIP. The iodoacetate-associated reduction in ADNP mRNA was prevented by pre-treatment with SNV. These effects imply that SNV provides a regulatory mechanism for ADNP synthesis during glycolytic stress. Furthermore, a short exposure to SNV provided potent protection from iodoacetate-induced toxicity suggesting that SNV may have therapeutic value in the treatment of ischemic/hypoxic injury. C1 Tel Aviv Univ, Sackler Sch Med, Dept Clin Biochem, IL-69978 Tel Aviv, Israel. Weizmann Inst Sci, Dept Organ Chem, IL-76100 Rehovot, Israel. NICHHD, Dev Neurobiol Lab, Sect Dev & Mol Pharmacol, NIH, Bethesda, MD 20892 USA. RP Sigalov, E (reprint author), Tel Aviv Univ, Sackler Sch Med, Dept Clin Biochem, IL-69978 Tel Aviv, Israel. NR 36 TC 49 Z9 51 U1 1 U2 1 PU HUMANA PRESS INC PI TOTOWA PA 999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USA SN 0895-8696 J9 J MOL NEUROSCI JI J. Mol. Neurosci. PD DEC PY 2000 VL 15 IS 3 BP 147 EP 154 DI 10.1385/JMN:15:3:147 PG 8 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 415XR UT WOS:000167749300002 PM 11303779 ER PT J AU Contreras, MA Greiner, RS Chang, MCJ Myers, CS Salem, N Rapoport, SI AF Contreras, MA Greiner, RS Chang, MCJ Myers, CS Salem, N Rapoport, SI TI Nutritional deprivation of alpha-linolenic acid decreases but does not abolish turnover and availability of unacylated docosahexaenoic acid and docosahexaenoyl-CoA in rat brain SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE docosahexaenoic acid; alpha-linolenic acid; phospholipids; polyunsaturated fatty acids; brain recycling; diet ID N-3 FATTY-ACIDS; SIGNAL-TRANSDUCTION; PHOSPHOLIPASE A(2); NERVOUS-SYSTEM; RHESUS-MONKEYS; DIETARY-FAT; IN-VIVO; DEFICIENCY; CELLS; METABOLISM AB We applied our in vivo fatty acid method to examine concentrations, incorporation, and turnover rates of docosahexaenoic acid (22:6 n-3) in brains of rats subject to a dietary deficiency of a-linolenic acid (18:3 n-3) for three generations. Adult deficient and adequate rats of the F3 generation were infused intravenously with [4,5-H-3]docosahexaenoic acid over 5 min, after which brain uptake and distribution of tracer were measured. Before infusion, the plasma 22:6 n-3 level was 0.2 nmol ml(-1) in 18:3 n3-deficient compared with 10.6 nmol ml(-1) in control rats. Brain unesterified 22:6 n-3 was not detectable, whereas docosahexaenoyl-CoA content was reduced by 95%, and 22:6 n-3 content in different phospholipid classes was reduced by 83-88% in deficient rats. Neither plasma or brain arachidonic acid (20:4 n-6) level was significantly changed with diet. Docosapentaenoic acid (22:5 n-6) reciprocally replaced 22:6 n-3 in brain phospholipids. Calculations using operational equations from our model indicated that 22:6 n-3 incorporation from plasma into brain was reduced 40-fold by 18:3 n-3 deficiency. Recycling of 22:6 n-3 due to deacylation-reacylation within phospholipids was reduced by 30-70% with the deficient diet, but animals nevertheless continued to produce 22:6 n-3 and docosahexaenoyl-CoA for brain function. We propose that functional brain effects of n-3 deficiency reflect altered ratios of n-6 to n-3 fatty acids. C1 NIA, Sect Brain Physiol & Metab, NIH, Bethesda, MD 20892 USA. NIAAA, Lab Membrane Biochem & Biophys, NIH, Bethesda, MD 20892 USA. RP NIA, Sect Brain Physiol & Metab, NIH, Bldg 10,6C-103,9000 Rockville Pike, Bethesda, MD 20892 USA. EM contrera@exchange.nih.gov NR 80 TC 82 Z9 83 U1 0 U2 0 PU WILEY PI HOBOKEN PA 111 RIVER ST, HOBOKEN 07030-5774, NJ USA SN 0022-3042 EI 1471-4159 J9 J NEUROCHEM JI J. Neurochem. PD DEC PY 2000 VL 75 IS 6 BP 2392 EP 2400 DI 10.1046/j.1471-4159.2000.0752392.x PG 9 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 373PY UT WOS:000165298900017 PM 11080190 ER PT J AU Di Marzo, V Breivogel, CS Tao, Q Bridgen, DT Razdan, RK Zimmer, AM Zimmer, A Martin, BR AF Di Marzo, V Breivogel, CS Tao, Q Bridgen, DT Razdan, RK Zimmer, AM Zimmer, A Martin, BR TI Levels, metabolism, and pharmacological activity of anandamide in CB1 cannabinoid receptor knockout mice: Evidence for non-CB1, non-CB2 receptor-mediated actions of anandamide in mouse brain SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE cannabinoid; anandamide; 2-arachidonyl glycerol; tetrahydrocannabinol; CB1 receptor knockout; C57BL/6 mouse ID CENTRAL-NERVOUS-SYSTEM; ACID AMIDE HYDROLASE; RAT-BRAIN; MOLECULAR CHARACTERIZATION; SELECTIVE ANTAGONIST; SR 141716A; OUT MICE; ENZYME; LIGAND; BINDS AB Anandamide [arachidonylethanolamide (AEA)] appears to be an endogenous agonist of brain cannabinoid receptors (CB1), yet some of the neurobehavioral effects of this compound in mice are unaffected by a selective CB1 antagonist. We studied the levels, pharmacological actions, and degradation of AEA in transgenic mice lacking the CB1 gene. We quantified AEA and the other endocannabinoid, 2-arachidonoyl glycerol, in six brain regions and the spinal cord by isotope-dilution liquid chromatography-mass spectrometry. The distribution of endocannabinoids and their inactivating enzyme, fatty acid amide hydrolase, were found to overlap with CB1 distribution only in part. In CB1 knockout homozygotes (CB1-/-), the hippocampus and, to a lesser extent, the striatum exhibited lower AEA levers as compared with wild-type (CB1+/+) controls. These data suggest a ligand/receptor relationship between AEA and CB1 in these two brain regions, where tonic activation of the receptor may tightly regulate the biosynthesis of its endogenous ligand. 2-Arachidonoyl glycerol levels and fatty acid amide hydrolase activity were unchanged in CB1-/- with respect to CB1 +/+ mice in all regions. AEA and Delta (9)-tetrahydrocannabinol (THC) were tested In CB1-/- mice for their capability of inducing analgesia and catalepsy and decreasing spontaneous activity. The effects of AEA, unlike THC, were not decreased in CB1-/- mice. AEA, but not THC, stimulated GTP gammaS binding in brain membranes from CB1-/- mice, and this stimulation was insensitive to CB1 and CB2 antagonists. We suggest that non-CB1, non-CB2 G protein-coupled receptors might mediate in mice some of the neurobehavioral actions of AEA. C1 Virginia Commonwealth Univ, Med Coll Virginia, Dept Pharmacol & Toxicol, Richmond, VA 23298 USA. CNR, Ist Chim Mol Interesse Biol, I-80072 Arco, Italy. Organix Inc, Woburn, MA USA. NIMH, Genet Lab, Bethesda, MD 20892 USA. RP Martin, BR (reprint author), Virginia Commonwealth Univ, Med Coll Virginia, Dept Pharmacol & Toxicol, POB 980613, Richmond, VA 23298 USA. RI Zimmer, Andreas/B-8357-2009; OI Breivogel, Chris/0000-0001-5657-2735; Di Marzo, Vincenzo/0000-0002-1490-3070 FU NIDA NIH HHS [DA 03672, DA 09789] NR 50 TC 296 Z9 305 U1 0 U2 4 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD DEC PY 2000 VL 75 IS 6 BP 2434 EP 2444 DI 10.1046/j.1471-4159.2000.0752434.x PG 11 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 373PY UT WOS:000165298900022 PM 11080195 ER PT J AU Moriguchi, T Greiner, RS Salem, N AF Moriguchi, T Greiner, RS Salem, N TI Behavioral deficits associated with dietary induction of decreased brain docosahexaenoic acid concentration SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE n-3 polyunsaturated fatty acid deficiency; docosahexaenoic acid; brain fatty acid composition; Morris water maze; learning ability; rat ID POLYUNSATURATED FATTY-ACIDS; LINOLENATE-LINOLEATE BALANCE; MORRIS WATER MAZE; VISUAL-ACUITY DEVELOPMENT; TERM INFANTS; LEARNING-ABILITY; LIPID COMPOSITIONS; ARACHIDONIC-ACID; PRETERM INFANTS; N-3 DEFICIENCY AB Docosahexaenoic acid (DHA), an n-3 fatty acid, is rapidly deposited during the period of rapid brain development. The influence of n-3 fatty acid deficiency on learning performance in adult rats over two generations was investigated. Rats were fed either an n-3 fatty acid-adequate (n-3 Adq) or -deficient (n-3 Def) diet for three generations (F1-F3). Levels of total brain n-3 fatty acids were reduced in the n-3 Def group by 83 and 87% in the F2 and F3 generations, respectively. In the Morris water maze, the n-3 Def group showed a longer escape latency and delayed acquisition of this task compared with the n-3 Adq group in both generations. The acquisition and memory levels of the n-3 Def group in the F3 generation seemed to be lower than that of the F2 generation. The 22:5n-6/22:6n-3 ratio in the frontal cortex and dams' milk was markedly increased in the n-3 Def group, and this ratio was significantly higher in the F3 generation compared with the F2 generation. These results suggest that learning and cognitive behavior are related to brain DHA status, which, in turn, is related to the levels of the milk/dietary n-3 fatty acids. C1 NIAAA, Lab Membrane Biochem & Biophys, NIH, Rockville, MD 20852 USA. RP Salem, N (reprint author), NIAAA, Lab Membrane Biochem & Biophys, NIH, 12420 Parklawn Dr,Room 158, Rockville, MD 20852 USA. NR 62 TC 255 Z9 268 U1 2 U2 14 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD DEC PY 2000 VL 75 IS 6 BP 2563 EP 2573 DI 10.1046/j.1471-4159.2000.0752563.x PG 11 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 373PY UT WOS:000165298900037 PM 11080210 ER PT J AU Gressens, P Arquie, C Hill, JM Marret, S Sahir, N Robberecht, P Evrard, P AF Gressens, P Arquie, C Hill, JM Marret, S Sahir, N Robberecht, P Evrard, P TI VIP and PACAP 38 modulate ibotenate-induced neuronal heterotopias in the newborn hamster neocortex SO JOURNAL OF NEUROPATHOLOGY AND EXPERIMENTAL NEUROLOGY LA English DT Article DE cAMP; ectopia; excitatory amino acids; N-methyl-D-aspartate; neuronal migration; RO 25-1553 ID VASOACTIVE-INTESTINAL-PEPTIDE; MURINE DEVELOPING BRAIN; DEVELOPING RAT-BRAIN; PROTEIN-KINASE-C; CEREBRAL-CORTEX; FUNCTIONAL EXPRESSION; EXCITOTOXIC LESIONS; CORTICAL-NEURONS; NMDA RECEPTORS; IN-VITRO AB Intracerebral administration of ibotenate produces, through activation of N-methyl-D-aspartate (NMDA) receptors, neuronal heterotopias in the newborn hamster neocortex: high doses of ibotenate induce periventricular and subcortical neuronal heterotopias, while low doses of ibotenate produce intracortical heterotopias and molecular layer ectopias. Vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) are closely related peptides with neurotrophic properties. They share common VPAC(1) and VPAC(2) receptors, which use cAMP as a second messenger. Previous studies have shown that VIP prevents excitotoxic neuronal death and exacerbates glutamate-induced c-fos neuronal expression. In order to gain new insight into the molecular control of neuronal migration, the present study examined the effects of VIP and PACAP on ibotenate-induced heterotopias in the newborn hamster. Go-treatment with VIP and a high dose of ibotenate produced a pattern of neuronal heterotopias similar to the one observed in animals treated with low doses of ibotenate alone. Pups co-injected with a low dose of ibotenate and a VIP antagonist displayed cortical dysgeneses similar to those observed in animals treated with high doses of ibotenate alone. The modulating effects of VIP on excitotoxin-induced heterotopias were mimicked by forskolin, PACAP, and by a specific VPAC(2) receptor agonist but not by a VPAC(1) agonist, and were blocked by a protein kinase A (PKA) inhibitor. Taken together, these data suggest that VIP and PAGAP can attenuate ibotenate-induced heterotopias in newborn hamster and that this effect is mediated by the VPAC(2) receptor utilizing the cAMP-PKA pathway. C1 Hop Robert Debre, Serv Neuropediat, INSERM E 9935, F-75019 Paris, France. Fac Med & Pharm Rouen, MERCI UPRESA 2122, Rouen, France. NICHD, Dev Neurobiol Lab, Sect Dev & Mol Pharmacol, Bethesda, MD USA. Univ Brussels, Sch Med, Lab Chim Biol & Nutr, Brussels, Belgium. RP Gressens, P (reprint author), Hop Robert Debre, Serv Neuropediat, INSERM E 9935, 48 Blvd Serurier, F-75019 Paris, France. NR 60 TC 16 Z9 17 U1 0 U2 0 PU AMER ASSN NEUROPATHOLOGISTS INC PI LAWRENCE PA 1041 NEW HAMPSHIRE ST, LAWRENCE, KS 66044 USA SN 0022-3069 J9 J NEUROPATH EXP NEUR JI J. Neuropathol. Exp. Neurol. PD DEC PY 2000 VL 59 IS 12 BP 1051 EP 1062 PG 12 WC Clinical Neurology; Neurosciences; Pathology SC Neurosciences & Neurology; Pathology GA 383XR UT WOS:000165910400004 PM 11138925 ER PT J AU Whelan, P Bonnot, A O'Donovan, MJ AF Whelan, P Bonnot, A O'Donovan, MJ TI Properties of rhythmic activity generated by the isolated spinal cord of the neonatal mouse SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID CENTRAL PATTERN GENERATORS; PAW-SHAKE RESPONSE; IN-VITRO; MOTOR PATTERNS; HINDLIMB LOCOMOTION; SCRATCH REFLEX; CHICK-EMBRYO; NEWBORN RAT; 3 FORMS; NMDA AB We examined the ability of the isolated lumbosacral spinal cord of the neonatal mouse (P0-7) to generate rhythmic motor activity under several different conditions. In the absence of electrical or pharmacological stimulation, we recorded several patterns of spontaneous ventral root depolarization and discharge. Spontaneous, alternating discharge between contralateral ventral roots could occur two to three times over a 10-min interval. We also observed other patterns, including left-right synchrony and rhythmic activity restricted to one side of the cord. Trains of stimuli delivered to the lumbar/coccygeal dorsal roots or the sural nerve reliably evoked episodes of rhythmic activity. During these evoked episodes, rhythmic ventral root discharges could occur on one side of the cord or could alternate from side to side. Bath application of a combination of N-methyl-D,L-aspartate (NMA), serotonin, and dopamine produced rhythmic activity that could last for several hours. Under these conditions, the discharge recorded from the left and right L-1-L-3 ventral roots alternated. In the L-4-L-5 segments, the discharge had two peaks in each cycle, coincident with discharge of the ipsilateral and contralateral L-1-L-3 roots. The L-6 ventral root discharge alternated with that recorded from the ipsilateral L-1-L-3 roots. We established that the drug-induced rhythm was locomotor-like by recording an alternating pattern of discharge between ipsilateral flexor and extensor hindlimb muscle nerves. In addition, by recording simultaneously from ventral roots and muscle nerves, we established that ankle flexor discharge was in phase with ipsilateral L-1/L-2 ventral root discharge, while extensor discharge was in phase with ipsilateral L6 ventral root discharge. Rhythmic patterns of ventral root discharge were preserved following mid-sagittal section of the spinal cord, demonstrating that reciprocal inhibitory connections between the left and right sides of the cord are not essential for rhythmogenesis in the neonatal mouse cord. Blocking N-methyl-D-aspartate receptors, in both the intact and the hemisected preparation, revealed that these receptors contribute to but are not essential for rhythmogenesis. In contrast, the rhythm was abolished following blockade of kainate/AMPA receptors with 6-cyano-7-nitroquinoxalene-2,3- dione. These findings demonstrate that the isolated mouse spinal cord can produce a variety of coordinated activities, including locomotor-like activity. The ability to study these behaviors under a variety of different conditions offers promise for future studies of rhythmogenic mechanisms in this preparation. C1 NINDS, Lab Neural Control, Sect Dev Neurobiol, NIH, Bethesda, MD 20892 USA. RP O'Donovan, MJ (reprint author), NINDS, Lab Neural Control, Sect Dev Neurobiol, NIH, Bldg 49,Rm 3A50, Bethesda, MD 20892 USA. RI o'donovan, michael/A-2357-2015 OI o'donovan, michael/0000-0003-2487-7547 NR 53 TC 157 Z9 159 U1 1 U2 5 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3077 J9 J NEUROPHYSIOL JI J. Neurophysiol. PD DEC PY 2000 VL 84 IS 6 BP 2821 EP 2833 PG 13 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 380TN UT WOS:000165719700011 PM 11110812 ER PT J AU Wilson, SM Toth, PT Oh, SB Gillard, SE Volsen, S Ren, DJ Philipson, LH Lee, EC Fletcher, CF Tessarollo, L Copeland, NG Jenkins, NA Miller, RJ AF Wilson, SM Toth, PT Oh, SB Gillard, SE Volsen, S Ren, DJ Philipson, LH Lee, EC Fletcher, CF Tessarollo, L Copeland, NG Jenkins, NA Miller, RJ TI The status of voltage-dependent calcium channels in alpha(1E) knock-out mice SO JOURNAL OF NEUROSCIENCE LA English DT Article DE dorsal root ganglia; cerebellar granule cells; pain; synaptic transmission; voltage-dependent calcium channels; alpha(1E) knock-out mice ID RAT CEREBELLAR NEURONS; SUBUNITS; CURRENTS; CELLS; ALPHA-1E; EXPRESSION; DIVERSITY; CLONING; MEMBER; FAMILY AB It has been hypothesized that R-type Ca currents result from the expression of the alpha (1E) gene. To test this hypothesis we examined the properties of voltage-dependent Ca channels in mice in which the alpha (1E) Ca channel subunit had been deleted. Application of omega -conotoxin GVIA, omega -agatoxin IVA, and nimodipine to cultured cerebellar granule neurons from wild-type mice inhibited components of the whole-cell Ba current, leaving a "residual" R current with an amplitude of similar to 30% of the total Ba current. A minor portion of this R current was inhibited by the alpha (1E)-selective toxin SNX-482, indicating that it resulted from the expression of alpha (1E). However, the majority of the R current was not inhibited by SNX-482. The SNX-482-sensitive portion of the granule cell R current was absent from alpha (1E) knock-out mice. We also identified a subpopulation of dorsal root ganglion (DRG) neurons from wild-type mice that expressed an SNX-482-sensitive component of the R current. However as with granule cells, most of the DRG R current was not blocked by SNX-482. We conclude that there exists a component of the R current that results from the expression of the alpha (1E) Ca channel subunit but that the majority of R currents must result from the expression of other Ca channel alpha subunits. C1 Univ Chicago, Dept Neurobiol Pharmacol & Physiol, Chicago, IL 60637 USA. NCI, Mouse Canc Genet Program, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. Eli Lilly & Co, Lilly Corp Ctr, Indianapolis, IN 46285 USA. Eli Lilly & Co, Windlesham GU20 6PH, Surrey, England. RP Miller, RJ (reprint author), Univ Chicago, Dept Neurobiol Pharmacol & Physiol, 947 E 58th St,MC 0926, Chicago, IL 60637 USA. FU NIDA NIH HHS [DA-02121]; NIMH NIH HHS [MH-40165]; NINDS NIH HHS [NS-33826] NR 27 TC 101 Z9 104 U1 0 U2 2 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD DEC 1 PY 2000 VL 20 IS 23 BP 8566 EP 8571 PG 6 WC Neurosciences SC Neurosciences & Neurology GA 378PD UT WOS:000165592000006 PM 11102459 ER PT J AU Trinidad, JC Fischbach, GD Cohen, JB AF Trinidad, JC Fischbach, GD Cohen, JB TI The agrin/MuSK signaling pathway is spatially segregated from the neuregulin/ErbB receptor signaling pathway at the neuromuscular junction SO JOURNAL OF NEUROSCIENCE LA English DT Article DE neuregulin; erbB receptor; agrin; MuSK; neuromuscular junction; immunohistochemistry ID EPSILON-SUBUNIT GENE; SKELETAL-MUSCLE FIBERS; ACETYLCHOLINE-RECEPTOR; POSTSYNAPTIC SPECIALIZATION; MYOGENIC DIFFERENTIATION; WALLERIAN DEGENERATION; PUTATIVE RECEPTORS; MAMMALIAN MUSCLE; SODIUM-CHANNELS; NERVOUS-SYSTEM AB The neuregulin/erbB receptor and agrin/MuSK pathways are critical for communication between the nerve, muscle, and Schwann cell that establishes the precise topological arrangement at the vertebrate neuromuscular junction (NMJ). ErbB2, erbB3, and erbB4 as well as neuregulin, agrin, and MuSK are known to be concentrated at the NMJ. Here we have examined NMJs from gastrocnemius muscle of adult rat using immunofluorescence confocal microscopy to characterize in detail the distribution of these proteins relative to the distribution of acetylcholine receptors (AChRs). We have determined that erbB2 and erbB4 are enriched in the depths of the secondary junctional folds on the postsynaptic muscle membrane. In contrast, erbB3 at the NMJ was concentrated at presynaptic terminal Schwann cells. This distribution strongly argues that erbB2/erbB4 heterodimers are the functional postsynaptic neuregulin receptors of the NMJ. Neuregulin was localized to the axon terminal, secondary folds, and terminal Schwann cells, where it was in a position to signal through erbB receptors. MuSK was concentrated in the postsynaptic primary gutter region where it was codistributed with AChRs. Agrin was present at the axon terminal and in the basal lamina associated with the primary gutter region, but not in the secondary junctional folds. The differential distributions of the neuregulin and agrin signaling pathways argue against neuregulin and erbB receptors being localized to the NMJ via direct interactions with either agrin or MuSK. C1 Harvard Univ, Sch Med, Dept Neurobiol, Boston, MA 02115 USA. NINDS, Bethesda, MD 20892 USA. RP Cohen, JB (reprint author), Harvard Univ, Sch Med, Dept Neurobiol, 220 Longwood Ave, Boston, MA 02115 USA. FU NINDS NIH HHS [NS18458] NR 58 TC 74 Z9 74 U1 0 U2 1 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD DEC 1 PY 2000 VL 20 IS 23 BP 8762 EP 8770 PG 9 WC Neurosciences SC Neurosciences & Neurology GA 378PD UT WOS:000165592000031 PM 11102484 ER PT J AU Vrij, A Edward, K Roberts, KP Bull, R AF Vrij, A Edward, K Roberts, KP Bull, R TI Detecting deceit via analysis of verbal and nonverbal behavior SO JOURNAL OF NONVERBAL BEHAVIOR LA English DT Article DE detecting deceit; nonverbal behavior; Criteria-Based Content Analysis; Reality Monitoring ID SEXUAL ABUSE; COGNITIVE INTERVIEW; FALSE STATEMENTS; POLICE OFFICERS; HAND MOVEMENTS; DECEPTION; CREDIBILITY; ALLEGATIONS; ABILITY; LIES AB We examined the hypotheses that (1) a systematic analysis of nonverbal behavior could be useful in the detection of deceit and (2) that lie detection would be most accurate ii both verbal and nonverbal indicators of deception are taken into account. Seventy-three nursing students participated in a study about "telling lies" and either told the truth or lied about a film they had just seen. The interviews were videotaped and audiotaped, and the nonverbal behavior (NVB) and speech content of the liars and truth tellers were analyzed, the latter with the Criteria-Based Content Analysis technique (CBCA) and the Reality Monitoring technique IRM). Results revealed several nonverbal and verbal indicators of deception. On the basis of nonverbal behavior alone, 78% of the lies and truths could be correctly classified. An even higher percentage could be correctly classified when all three detection techniques (i.e., NVB, CBCA, RM) were taken into account. C1 Univ Portsmouth, Dept Psychol, Portsmouth PO1 2DY, Hants, England. Natl Inst Child Hlth & Human Dev, Bethesda, MD USA. RP Vrij, A (reprint author), Univ Portsmouth, Dept Psychol, King Henry Bldg,King Henry 1 St, Portsmouth PO1 2DY, Hants, England. EM aldert.vrij@port.ac.uk NR 83 TC 135 Z9 137 U1 9 U2 49 PU SPRINGER PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 0191-5886 J9 J NONVERBAL BEHAV JI J. Nonverbal Behav. PD WIN PY 2000 VL 24 IS 4 BP 239 EP 263 DI 10.1023/A:1006610329284 PG 25 WC Psychology, Social SC Psychology GA 368VC UT WOS:000090137300001 ER PT J AU Govindan, SV Goldenberg, DM Elsamra, SE Griffiths, GL Ong, GL Brechbiel, MW Burton, J Sgouros, G Mattes, MJ AF Govindan, SV Goldenberg, DM Elsamra, SE Griffiths, GL Ong, GL Brechbiel, MW Burton, J Sgouros, G Mattes, MJ TI Radionuclides linked to a CD74 antibody as therapeutic agents for B-cell lymphoma: Comparison of Auger electron emitters with beta-particle emitters SO JOURNAL OF NUCLEAR MEDICINE LA English DT Article DE radiolabeled Abs; in vitro cytotoxicity; B-cell lymphoma; antiCD74 antibody ID MONOCLONAL-ANTIBODIES; INVARIANT CHAIN; IN-VITRO; SURFACE; INTERNALIZATION; BIODISTRIBUTION; TOXICITY; TUMORS; FATE AB We demonstrated previously that human B-cell lymphomas were effectively and specifically killed in vitro by an antibody to CD74 (LL1) linked to (111)ln or other Auger electron emitters. This study was intended to more accurately compare the potency and specificity of 3 Auger electron emitters, (111)ln, Ga-67, and I-125, and to evaluate beta -particle emitters, I-131 and Y-90. The unique property of LL1 is its high level of intracellular uptake. Methods: Raji B-lymphoma cells were incubated with serial dilutions of the radiolabeled Abs for 2 d and then monitored for cell growth by 2 assays: a cell counting assay and a clonogenic assay. The uptake of radioactivity per cell was monitored at various time points, and the radiation dose was calculated using published S values for radioactivity located in the cytoplasm. Both specific and nonspecific toxicity were evaluated. Results: The beta -particle emitters had considerably higher levels of nonspecific toxicity than the Auger electron emitters, but both I-131 and Y-90, and particularly I-131, still had high levels of specificity. Both of these results were consistent with dosimetry calculations. Relative to the delivered disintegrations per cell, I-131 and Ga-67 were the most potent of the radionuclides tested, with I-125 and In-111 being significantly weaker and Y-90 being intermediate. The high potency of Ga-67, together with its low nonspecific toxicity, caused this radionuclide to have the highest specificity index. Conclusion: When delivered by Ab LL1, both Auger electron and beta -particle emitters can produce specific and effective toxicity. The choice of the optimal radionuclide for therapy may depend on the ease and efficiency of labeling, the specific activity obtained, the nature of the tumor being targeted, and other factors, but the high specificity indices of the Auger electron emitters may be an advantage. C1 Ctr Mol Med & Immunol, Garden Canc Ctr, Belleville, NJ 07109 USA. Immuinomed Inc, Morris Plains, NJ USA. NIH, Bethesda, MD 20892 USA. Mem Sloan Kettering Canc Ctr, New York, NY 10021 USA. RP Mattes, MJ (reprint author), Ctr Mol Med & Immunol, Garden Canc Ctr, 520 Belleville Ave, Belleville, NJ 07109 USA. FU NCI NIH HHS [CA39841, CA72324] NR 22 TC 51 Z9 51 U1 0 U2 5 PU SOC NUCLEAR MEDICINE INC PI RESTON PA 1850 SAMUEL MORSE DR, RESTON, VA 20190-5316 USA SN 0161-5505 J9 J NUCL MED JI J. Nucl. Med. PD DEC PY 2000 VL 41 IS 12 BP 2089 EP 2097 PG 9 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 386NG UT WOS:000166068000030 PM 11138697 ER PT J AU Greenwald, P Milner, JA Clifford, CK AF Greenwald, P Milner, JA Clifford, CK TI Creating a new paradigm in nutrition research within the National Cancer Institute SO JOURNAL OF NUTRITION LA English DT Article DE nutrition; cancer research; paradigm; collaboration; training ID PREVENTION; DIET; GENE AB Almost two decades after Doll and Peto (1981) provided evidence that one third of cancer deaths are related to diet, it remains unclear which dietary components may be key in cancer prevention. Although the complexity of the diet can become overwhelming, the National Cancer Institute (NCl) of the National Institutes of Health (NIH) has remained steadfast in its commitment to defining the roles that diet and nutrition have in the development of cancer and has provided increased research and training support to assist in unraveling this interrelationship. Evidence for this sustained commitment is highlighted by a fourfold increase in NCI expenditures for nutrition research and training from 1983 to 1998; this substantial increase reflects a trend that is occurring in some universities and the private sector. More than one third of the nutrition-related NCI research is funded by the Division of Cancer Prevention. Supported investigations cover the gamut from basic mechanisms of action of dietary constituents, methodology development, human metabolic studies, clinical trials of dietary modification and the chemopreventive potential of individual nutrients to population-based studies. C1 NCI, Div Canc Prevent, NIH, Bethesda, MD 20892 USA. RP Greenwald, P (reprint author), NCI, Div Canc Prevent, NIH, Bethesda, MD 20892 USA. EM pg37g@nih.gov NR 13 TC 6 Z9 7 U1 0 U2 1 PU AMER SOC NUTRITIONAL SCIENCE PI BETHESDA PA 9650 ROCKVILLE PIKE, RM L-2407A, BETHESDA, MD 20814 USA SN 0022-3166 J9 J NUTR JI J. Nutr. PD DEC PY 2000 VL 130 IS 12 BP 3103 EP 3105 PG 3 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 383CU UT WOS:000165866200040 PM 11110877 ER PT J AU Hewitt, RE McMarlin, A Kleiner, D Wersto, R Martin, P Tsoskas, M Stamp, GWH Stetler-Stevenson, WG AF Hewitt, RE McMarlin, A Kleiner, D Wersto, R Martin, P Tsoskas, M Stamp, GWH Stetler-Stevenson, WG TI Validation of a model of colon cancer progression SO JOURNAL OF PATHOLOGY LA English DT Article DE metastasis; colon; cell line; xenograft ID CARCINOMA CELL-LINES; GROWTH-FACTOR; IN-VIVO; COLORECTAL-CANCER; EXPRESSION; METASTASIS; MODULATION; INVASION AB A unique feature of SW480 and SW620 colon carcinoma cell lines is that they are derived from primary and secondary tumours resected from a single patient. As such, they may represent a valuable resource for examining genetic changes late in colon cancer progression. In order to verify this, both cell lines have been characterized to determine whether phenotypic differences have been retained despite long-term cell culture in vitro. The primary tumour-derived SW480 cells have an epithelioid morphology in vitro, while metastasis-derived SW620 cells have a fibroblast-like appearance, Xenografts of SW480 cells form gland-like structures irt vivo, while SW620 xenografts form solid sheets of tumour cells, SW620 cells have a higher BrdU labelling index than SW480 cells, and are more highly tumourigenic and metastatic, Furthermore, SW620 cells show less susceptibility to apoptosis induction by TNF alpha and anti-Fas monoclonal, Findings from these investigations therefore indicate that SW480 and SW620 cell lines do show appropriate phenotypic differences and represent an interesting model for studying the genetic events in the late stages of colon cancer progression. Copyright (C) 2000 John Wiley & Sons, Ltd. C1 NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. Hammersmith Hosp, Dept Histopathol, London W12 0NN, England. RP Hewitt, RE (reprint author), King Faisal Specialist Hosp & Res Ctr, MBC 03,POB 3354, Riyadh 11211, Saudi Arabia. RI Stetler-Stevenson, William/H-6956-2012; OI Stetler-Stevenson, William/0000-0002-5500-5808; Kleiner, David/0000-0003-3442-4453 NR 18 TC 112 Z9 114 U1 3 U2 8 PU JOHN WILEY & SONS LTD PI W SUSSEX PA BAFFINS LANE CHICHESTER, W SUSSEX PO19 1UD, ENGLAND SN 0022-3417 J9 J PATHOL JI J. Pathol. PD DEC PY 2000 VL 192 IS 4 BP 446 EP 454 PG 9 WC Oncology; Pathology SC Oncology; Pathology GA 380KE UT WOS:000165700400005 PM 11113861 ER PT J AU Hewitt, RE Brown, KE Corcoran, M Stetler-Stevenson, WG AF Hewitt, RE Brown, KE Corcoran, M Stetler-Stevenson, WG TI Increased expression of tissue inhibitor of metalloproteinases type I (TIMP-I) in a more tumourigenic colon cancer cell line SO JOURNAL OF PATHOLOGY LA English DT Article DE metastasis; colon; cell line; differential display ID COLORECTAL-CANCER; GROWTH-FACTOR; CARCINOMA CELLS; GENE; MUTATIONS; VIVO; TUMORIGENICITY; RESISTANCE; MODULATION; METASTASIS AB Genetic changes occurring in the late stages of colonic tumour progression have received much less attention than those occurring in the early stages, As described in the accompanying paper, SW480 and SW620 cell lines provide a useful model for studying the advanced stages of progression for colon cancer. Comparison of the two cell lines by differential display reveals that SW620 cells express lower levels of the CC3 tumour suppressor gene and also lower levels of the tissue inhibitor of metalloproteinases-3 (TIMP-3) gene. Northern blot analysis for TIMP-3 confirms this finding and shows a similar difference in the expression of TIMP-2, which seems logical since TIMPs inhibit enzymes that play a role in tumour invasion. For this reason, it was surprising to find that TIMP-1 messenger RNA expression is markedly increased in SW620 cells, Consistent with this finding, western blot analysis shows a ten-fold increase in TIMP-1 protein secretion by SW620 cells, It is noteworthy that high TIMP-1 expression is associated with poor prognosis in colorectal cancer. This association between TIMP-1 expression and tumour progression may be related to additional growth factor-like effects described for TIMP-1 in some systems. Copyright (C) 2000 John Wiley & Sons, Ltd. C1 NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. NHLBI, Dept Hematol, NIH, Bethesda, MD 20892 USA. RP Hewitt, RE (reprint author), King Faisal Specialist Hosp & Res Ctr, MBC 03,POB 3354, Riyadh 11211, Saudi Arabia. RI Stetler-Stevenson, William/H-6956-2012 OI Stetler-Stevenson, William/0000-0002-5500-5808 NR 32 TC 41 Z9 43 U1 0 U2 0 PU JOHN WILEY & SONS LTD PI W SUSSEX PA BAFFINS LANE CHICHESTER, W SUSSEX PO19 1UD, ENGLAND SN 0022-3417 J9 J PATHOL JI J. Pathol. PD DEC PY 2000 VL 192 IS 4 BP 455 EP 459 PG 5 WC Oncology; Pathology SC Oncology; Pathology GA 380KE UT WOS:000165700400006 PM 11113862 ER PT J AU Yanovski, JA Sovik, KN Nguyen, TT Sebring, NG AF Yanovski, JA Sovik, KN Nguyen, TT Sebring, NG TI Insulin-like growth factors and bone mineral density in African American and white girls SO JOURNAL OF PEDIATRICS LA English DT Article ID CLINICAL RESEARCH-CENTER; HORMONE GH TREATMENT; FACTOR-I; RACIAL-DIFFERENCES; PREMENOPAUSAL WOMEN; CALCIUM-ABSORPTION; HEALTHY-CHILDREN; BINDING-PROTEIN; LARON SYNDROME; FEMORAL-NECK AB Objectives: African American children have greater bone mineral density (BMD) and bone mineral content (BMC) than white children. We examined the hypothesis that differences in insulin-like growth factors (IGFs) are important determinants of BMD during childhood. Methods: We measured IGFs and IGF binding proteins in 59 African American and 59 white girls matched for age, body mass index, socioeconomic status, and pubertal stage. BMD and BMC were determined by dual emission x-ray absorptiometry, Results: African American girls had greater total BMD (P < .001), BMC (P < .01), total IGF-1 (P < .001),and free IGF-1 (P < .01) than white girls. IGFBP-1, IGFBP-2, and IGFBP-3 were similar in both groups or lower in African Americans. IGF-1 was positively correlated with IGF-2 in white girls (P = .012) but was negatively correlated with IGF-2 in African Americans (P = .015). IGF-1 and free IGF-1 were positively correlated with BMD/BMC. Multiple regression analyses showed 80% of the variance in BMC could be accounted for by the use of body weight, height, and IGF-1 in the model. When IGF-1 was included as a factor, race did not add to the model's predictive power. Conclusion: IGF-1 and free IGF-1 are greater in African American than in white girls and may contribute to the greater BMD of African Americans. C1 NICHHD, Unit Growth & Obes, Dev Endocrinol Branch, Div Nutr Res Coordinat,NIH, Bethesda, MD 20892 USA. NIH, Dept Nutr, Bethesda, MD 20892 USA. NIH, Warren Grant Magnuson Clin Ctr, Bethesda, MD 20892 USA. RP Yanovski, JA (reprint author), NICHHD, Unit Growth & Obes, Dev Endocrinol Branch, Div Nutr Res Coordinat,NIH, Bldg 10,Room 10N262,10 Ctr Dr,MSC 1862, Bethesda, MD 20892 USA. OI Yanovski, Jack/0000-0001-8542-1637 FU NICHD NIH HHS [ZO1-HD-00641] NR 48 TC 35 Z9 36 U1 0 U2 0 PU MOSBY-ELSEVIER PI NEW YORK PA 360 PARK AVENUE SOUTH, NEW YORK, NY 10010-1710 USA SN 0022-3476 EI 1097-6833 J9 J PEDIATR-US JI J. Pediatr. PD DEC PY 2000 VL 137 IS 6 BP 826 EP 832 DI 10.1067/mpd.2000.109151 PG 7 WC Pediatrics SC Pediatrics GA 383HD UT WOS:000165876300016 PM 11113840 ER PT J AU Albandar, JM Streckfus, CE Adesanya, MR Winn, DM AF Albandar, JM Streckfus, CE Adesanya, MR Winn, DM TI Cigar, pipe, and cigarette smoking as risk factors for periodontal disease and tooth loss SO JOURNAL OF PERIODONTOLOGY LA English DT Article DE periodontal diseases, etiology; smoking, adverse effects; smoking cessation; tooth loss, etiology; risk factors ID ALVEOLAR BONE LOSS; ADULTS 30 YEARS; UNITED-STATES; EXPERIMENTAL GINGIVITIS; LUNG-CANCER; TOBACCO USE; SMOKERS; ASSOCIATION; EXPRESSION; PREVALENCE AB Background: Our purpose was to test the hypotheses that cigar and pipe smoking have significant associations with periodontal disease and cigar, pipe, and cigarette smoking is associated with tooth loss. We also investigated whether a history of smoking habits cessation may affect the risk of periodontal disease and tooth loss. Methods: A group of 705 individuals (21 to 92 years-old) who were among volunteer participants in the ongoing Baltimore Longitudinal Study of Aging were examined clinically to assess their periodontal status and tooth loss. A structured interview was used to assess the participants' smoking behaviors with regard to cigarettes, cigar, and pipe smoking status. For a given tobacco product, current smokers were defined as individuals who at the time of examination continued to smoke daily. Former heavy smokers were defined as individuals who have smoked daily for 10 or more years and who had quit smoking. Non-smokers included individuals with a previous history of smoking for less than 10 years or no history of smoking. Results: Cigarette and cigar/pipe smokers had a higher prevalence of moderate and severe periodontitis and higher prevalence and extent of attachment loss and gingival recession than non-smokers, suggesting poorer periodontal health in smokers. In addition, smokers had less gingival bleeding and higher number of missing teeth than non-smokers. Current cigarette smokers had the highest prevalence of moderate and severe periodontitis (25.7%) compared to former cigarette smokers (20.2%), and non-smokers (13.1%). The estimated prevalence of moderate and severe periodontitis in current or former cigar/pipe smokers was 17,6%. A similar pattern was seen for other periodontal measurements including the percentages of teeth with greater than or equal to5 mm attachment loss and probing depth, greater than or equal to3 mm gingival recession, and dental calculus. Current, former, and non- cigarette smokers had 5.1, 3,9, and 2.8 missing teeth, respectively. Cigar/pipe smokers had on average 4 missing teeth. Multiple regression analysis also showed that current tobacco smokers may have increased risks of having moderate and severe periodontitis than former smokers. However, smoking behaviors explained only small percentages (<5%) of the variances in the multivariate models. Conclusion: The results suggest that cigar and pipe smoking may have similar adverse effects on periodontal health and tooth loss as cigarette smoking. Smoking cessation efforts should be considered as a means of improving periodontal health and reducing tooth loss in heavy smokers of cigarettes, cigars, and pipes with periodontal disease. C1 Temple Univ, Sch Dent, Dept Periodontol, Philadelphia, PA 19140 USA. Univ Mississippi, Sch Dent, Dept Diagnost Sci, Jackson, MS 39216 USA. Natl Inst Dent & Craniofacial Res, Div Intramural Res, Bethesda, MD USA. RP Albandar, JM (reprint author), Temple Univ, Sch Dent, Dept Periodontol, 3223 N Broad St, Philadelphia, PA 19140 USA. OI Albandar, Jasim M./0000-0001-7801-3811 NR 32 TC 151 Z9 155 U1 1 U2 16 PU AMER ACAD PERIODONTOLOGY PI CHICAGO PA 737 NORTH MICHIGAN AVENUE, SUITE 800, CHICAGO, IL 60611-2690 USA SN 0022-3492 J9 J PERIODONTOL JI J. Periodont. PD DEC PY 2000 VL 71 IS 12 BP 1874 EP 1881 DI 10.1902/jop.2000.71.12.1874 PG 8 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA 411GT UT WOS:000167490200007 PM 11156044 ER PT J AU Trobst, KK Wiggins, JS Costa, PT Herbst, JH McCrae, RR Masters, HL AF Trobst, KK Wiggins, JS Costa, PT Herbst, JH McCrae, RR Masters, HL TI Personality psychology and problem behaviors: HIV risk and the five-factor model SO JOURNAL OF PERSONALITY LA English DT Article ID 5-FACTOR MODEL; SEXUAL-BEHAVIOR; AIDS; HEALTH; VULNERABILITY; PERCEPTIONS; PREVENTION; INFECTION; OPTIMISM AB Studies of personality and problem behaviors may begin with analyses of the problem and develop hypotheses about personality traits that might be relevant; or they may begin with models of personality and explore links to behavior. Because it is well validated and relatively comprehensive, the Five-Factor Model (FFM) of personality lends itself to systematic exploratory studies that may sometimes lead to unanticipated findings. In this article, we review a program of research in a high-risk, disadvantaged population that illustrates the utility of the FFM in understanding health risk behavior. Previous analyses showed that behavior associated with the risk of HIV infection can be predicted from the personality dispositions of Neuroticism and (low) Conscientiousness. C1 NIA, Gerontol Res Ctr, Lab Personal & Cognit, NIH, Baltimore, MD 21224 USA. HealthSmart PA, Little Rock, AR USA. RP Costa, PT (reprint author), NIA, Gerontol Res Ctr, Lab Personal & Cognit, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. OI Costa, Paul/0000-0003-4375-1712 NR 38 TC 39 Z9 39 U1 0 U2 7 PU BLACKWELL PUBLISHERS PI MALDEN PA 350 MAIN STREET, STE 6, MALDEN, MA 02148 USA SN 0022-3506 J9 J PERS JI J. Pers. PD DEC PY 2000 VL 68 IS 6 BP 1233 EP 1252 DI 10.1111/1467-6494.00133 PG 20 WC Psychology, Social SC Psychology GA 381BF UT WOS:000165741900012 PM 11130739 ER PT J AU Kling, MA Carson, RE Borg, L Zametkin, A Matochik, JA Schluger, J Herscovitch, P Rice, KC Ho, A Eckelman, WC Kreek, MJ AF Kling, MA Carson, RE Borg, L Zametkin, A Matochik, JA Schluger, J Herscovitch, P Rice, KC Ho, A Eckelman, WC Kreek, MJ TI Opioid receptor imaging with positron emission tomography and [(18)F]cyclofoxy in long-term, methadone-treated former heroin addicts SO JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS LA English DT Article ID MESSENGER-RNA LEVELS; MAINTENANCE TREATMENT; F-18 CYCLOFOXY; BINDING-SITES; BRAIN; PET; STRESS; PLASMA AB Stabilized methadone-maintained former heroin addicts (MTPs) treated with effective doses of methadone have markedly reduced drug craving; reduction or elimination of heroin use; normalized stress-responsive hypothalamic-pituitary-adrenal, reproductive, and gastrointestinal function; and marked improvement in immune function and normal responses to pain, all of which are physiological indices modulated in part by endogenous and exogenous opioids directed at the mu and, in some cases, the kappa-opioid systems. This study was performed to explore opioid receptor binding in MTPs. Fourteen normal, healthy volunteers and 14 long-term MTPs in treatment for 2 to 27 years and receiving 30 to 90 mg/day of methadone were studied with positron emission tomography using tracer amounts of [(18)F]cyclofoxy, an opioid antagonist that labels mu and kappa opioid receptors. Imaging was performed in the morning, 22 h after the last dose of methadone in patients, and concurrent plasma levels of methadone were determined. Five brain regions of specific interest for addiction and pain research (thalamus, amygdala, caudate, anterior cingulate cortex, and putamen) were among the six regions of highest [(18)F]cyclofoxy binding. Specific binding of [(18)F]cyclofoxy was lower by 19 to 32% in these regions in MTPs compared with those in normal volunteers. The degree to which specific binding was lower in caudate and putamen correlated with methadone plasma levels (P < .01 and P < .05, respectively), suggesting that these lower levels of binding may be related to receptor occupancy with methadone and that significant numbers of opioid receptors may be available to function in their normal physiological roles. C1 Rockefeller Univ, Lab Biol Addict Dis, New York, NY 10021 USA. NIDDKD, NIH, Bethesda, MD 20892 USA. NIMH, Ctr Clin, PET Dept, NIH, Bethesda, MD 20892 USA. NIDA, NIH, Baltimore, MD USA. RP Kreek, MJ (reprint author), Rockefeller Univ, Lab Biol Addict Dis, 1230 York Ave,Box 171, New York, NY 10021 USA. EM Russos@rockvax.rockefeller.edu RI Kling, Mitchel/F-4152-2010; Carson, Richard/H-3250-2011 OI Kling, Mitchel/0000-0002-2232-1409; Carson, Richard/0000-0002-9338-7966 FU NCRR NIH HHS [M01RR00102]; NIDA NIH HHS [DA-K05-00049, DAP50-05130] NR 36 TC 78 Z9 84 U1 1 U2 5 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3995 USA SN 0022-3565 J9 J PHARMACOL EXP THER JI J. Pharmacol. Exp. Ther. PD DEC PY 2000 VL 295 IS 3 BP 1070 EP 1076 PG 7 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 374HD UT WOS:000165339000026 PM 11082442 ER PT J AU Gasior, M Jaszyna, M Peters, J Goldberg, SR AF Gasior, M Jaszyna, M Peters, J Goldberg, SR TI Changes in the ambulatory activity and discriminative stimulus effects of psychostimulant drugs in rats chronically exposed to caffeine: Effect of caffeine dose SO JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS LA English DT Article ID LOCOMOTOR-ACTIVITY; PHYSICAL-DEPENDENCE; D-AMPHETAMINE; COCAINE; NICOTINE; ADENOSINE; HUMANS; BRAIN; PRETREATMENT; ACQUISITION AB Caffeine is a common psychoactive constituent of coffee, carbonated beverages, and over-the-counter medications. This study examined the effects of chronic caffeine exposure on the behavioral response to acute administrations of psychostimulant drugs on ambulatory activity and on the pharmacological characteristics of nicotine discrimination in rats. Rats were maintained continuously on caffeine added to the drinking water at a concentration of 0.25 or 1.0 mg/ml that resulted in plasma caffeine concentrations ranging from 0.37 to 5.95 mug/ml. Rats maintained on tap water served as control groups. Exposure to the lower caffeine concentration (0.25 mg/ml) potentiated stimulatory effects of nicotine, amphetamine, and cocaine on ambulatory activity and failed to produce tolerance to the acute stimulatory effects of caffeine. In contrast, exposure to the higher caffeine concentration (1.0 mg/ml) did not alter the effects of the psychomotor stimulants on ambulatory behaviors but resulted in the development of complete, insurmountable tolerance to the acute stimulatory effects of caffeine. In the nicotine discrimination paradigm (0.4 mg/kg, training dose, a fixed-ratio 10 schedule of food delivery in a two-lever choice paradigm), rats exposed to the lower, but not to the higher, caffeine concentration acquired the nicotine discrimination significantly faster and were more sensitive to the effects of amphetamine and cocaine in substitution tests than water-drinking rats. Caffeine exposure did not change pharmacokinetic properties of nicotine (i.e., plasma levels, metabolism). In summary, exposure to two different caffeine solutions within a range of plasma levels observed in humans resulted in quantitatively distinct changes in psychostimulant-induced nonoperant and operant measures of behavior. These results suggest that dietary consumption of moderate doses of caffeine may be associated with enhanced reactions to some psychostimulants. C1 NIDA, Preclin Pharmacol Lab, Intramural Res Program, NIH, Baltimore, MD USA. Med Univ Sch, Dept Pharmacol, Lublin, Poland. Med Univ Sch, Dept Internal Med, Lublin, Poland. RP Gasior, M (reprint author), Harvard Univ, McLean Hosp, Sch Med,Alcohol & Drug Abuse Res Ctr, Dept Psychiat,Behav Pharmacol Program, 115 Mill St, Belmont, MA 02178 USA. RI Peters, Jamie/B-4288-2013 NR 42 TC 52 Z9 54 U1 1 U2 5 PU AMER SOC PHARMACOLOGY EXPERIMENTAL THERAPEUTICS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0022-3565 J9 J PHARMACOL EXP THER JI J. Pharmacol. Exp. Ther. PD DEC PY 2000 VL 295 IS 3 BP 1101 EP 1111 PG 11 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 374HD UT WOS:000165339000030 PM 11082446 ER EF