FN Thomson Reuters Web of Science™ VR 1.0 PT J AU Ross, PD Rekharsky, MV AF Ross, PD Rekharsky, MV TI Thermodynamics of hydrogen bond and hydrophobic interactions in cyclodextrin complexes SO BIOPHYSICAL JOURNAL LA English DT Article ID PROTEIN-FOLDING THERMODYNAMICS; ALPHA-CYCLODEXTRIN; INCLUSION COMPLEXES; BETA-CYCLODEXTRIN; AQUEOUS-SOLUTION; HEAT-CAPACITIES; ENTHALPIES; TITRATION; MOLECULES; HYDRATION AB Values of K, Delta G degrees, Delta H degrees, Delta S degrees and Delta C-p degrees for the binding reaction of small organic ligands forming 1:1 complexes with either alpha- Or beta-cyclodextrin were obtained by titration calorimetry from 15 degrees C to 45 degrees C. A hydrogen bond or hydrophobic interaction was introduced by adding a single functional group to the ligand. The thermodynamics of binding with and without the added group are compared to estimate the contribution of the hydrogen bond or hydrophobic interaction. A change in the environment of a functional group is required to influence the binding thermodynamics, but molecular size-dependent solute-solvent interactions have no effect. For phenolic O-H-O hydrogen bond formation, Delta H degrees Varies from -2 to -1.4 kcal mol(-1) from 15 degrees C to 45 degrees C and Delta C-p degrees is increased by 18 cal K-1 mol(-1). The hydrophobic interaction has an opposite effect: in alpha-cyclodextrin, Delta C-p degrees = -13.3 cal K-1 mol(-1) per ligand -CH2-, identical-to values found for the transfer of a -CH2- group from water to a nonpolar environment. At room temperature, the hydrogen bond and the -CH2- interaction each contribute about -600 cal mol(-1) to the stability (Delta G degrees) of the complex, With increased temperature, the hydrogen bond stability decreases (i.e., hydrogen bonds ''melt''), but the stability of the hydrophobic interaction remains essentially constant. RP Ross, PD (reprint author), NIDDKD,MOL BIOL LAB,NIH,BLDG 5,RM 327,9000 ROCKVILLE PIKE,BETHESDA,MD 20892, USA. NR 40 TC 166 Z9 167 U1 1 U2 28 PU BIOPHYSICAL SOCIETY PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 SN 0006-3495 J9 BIOPHYS J JI Biophys. J. PD OCT PY 1996 VL 71 IS 4 BP 2144 EP 2154 PG 11 WC Biophysics SC Biophysics GA VK295 UT WOS:A1996VK29500047 PM 8889190 ER PT J AU Liu, KL Sasisekharan, V Miles, HT Raghunathan, G AF Liu, KL Sasisekharan, V Miles, HT Raghunathan, G TI Structure of Py center dot Pu center dot Py DNA triple helices. Fourier transforms of fiber-type x-ray diffraction of single crystals SO BIOPOLYMERS LA English DT Article ID SYMMETRY; D(T)N.D(A)N.D(T)N; DYNAMICS; MODELS; DUPLEX AB Well-formed hexagonal crystals of oligomeric DNA triple helices exhibit fiber-type x-ray diffraction patterns [cf., Lilt et al. (1994) Nature Struct. Biol. I, II], which can be interpreted in terms of Fourier transforms of these helices. Precession photographs of a triplex formed of DA and dr chains show that it has 13 residues per trim. In contrast, a sequence containing the four natural bases A, G, C, and T has 12 residues per turn. In this sense the triple helices exhibit a sequence-dependent polymorphism, though both have C2'-endo sugar pucker and B rather than A conformation. New models are constructed, using constraints from x-ray diffraction, and Fourier transforms of the models are calculated. Good agreement in the amplitudes and positions of the calculated and observed diffraction intensities confirms the structures for both triple helices. These are the first stereochemically satisfactory DNA triple helices for which coordinates based on adequate experimental data were provided. Sequences for crystallization are designed to achieve unique base alignments and are screened for the presence of sharp bands on gel electrophoresis to assure the absence of multiple species caused by strand slippage. Despite intensive efforts to observe normal crystal diffraction by varying sequences and conditions, all crystals exhibited only fiber-type diffraction. We suggest that this behavior may be an intrinsic property of triple helices and discuss possible reasons for the results. Spectroscopic and chemical experiments establish that the oligonucleotides exist in solution as triple helices under the conditions of crystallization. (C) 1996 John Wiley & Sons, Inc. C1 NCI,NIH,MATH BIOL LAB,BETHESDA,MD 20892. INST PHARMACOL & TOXICOL,BEIJING 100850,PEOPLES R CHINA. RP Liu, KL (reprint author), NIDDKD,MOL BIOL LAB,NIH,BETHESDA,MD 20892, USA. NR 36 TC 17 Z9 17 U1 0 U2 3 PU JOHN WILEY & SONS INC PI NEW YORK PA 605 THIRD AVE, NEW YORK, NY 10158-0012 SN 0006-3525 J9 BIOPOLYMERS JI Biopolymers PD OCT PY 1996 VL 39 IS 4 BP 573 EP 589 DI 10.1002/(SICI)1097-0282(199610)39:4<573::AID-BIP8>3.0.CO;2-U PG 17 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA VG942 UT WOS:A1996VG94200008 PM 8837521 ER PT J AU Magnuson, VL Ally, DS Nylund, SJ Karanjawala, ZE Rayman, JB Knapp, JI Lowe, AL Ghosh, S Collins, FS AF Magnuson, VL Ally, DS Nylund, SJ Karanjawala, ZE Rayman, JB Knapp, JI Lowe, AL Ghosh, S Collins, FS TI Substrate nucleotide-determined non-templated addition of adenine by Taq DNA polymerase: Implications for PCR-based genotyping and cloning SO BIOTECHNIQUES LA English DT Article ID PRODUCTS; VECTORS AB The Applied Biosystems PRISM(TM) florescence-based genotyping system as well as the Invitrogen TA Cloning(R) vector system are influenced by the tendency of Taq DNA polymerase to add an adenine nucleotide to the 3' end of PCR products after extension. Incomplete addition of adenine to a majority of PCR product strands creates problems in allele-calling during genotyping and potentially diminishes the cloning efficiency of such products. Experiments reported here show that certain terminal nucleotides can either inhibit or enhance adenine addition by Taq and that PCR primer design can be used to modulate this activity. The methods we propose can substantially improve allele-calling for problematic microsatellite markers when using GENOTYPER(TM) software. C1 PERKIN ELMER CORP,APPL BIOSYST DIV,FOSTER CITY,CA. RP Magnuson, VL (reprint author), NIH,NATL CTR HUMAN GENOME RES,LAB GENE TRANSFER,49 CONVENT DR,ROOM 3A-14,BETHESDA,MD 20892, USA. NR 9 TC 117 Z9 122 U1 2 U2 10 PU EATON PUBLISHING CO PI NATICK PA 154 E. CENTRAL ST, NATICK, MA 01760 SN 0736-6205 J9 BIOTECHNIQUES JI Biotechniques PD OCT PY 1996 VL 21 IS 4 BP 700 EP 709 PG 10 WC Biochemical Research Methods; Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VL405 UT WOS:A1996VL40500024 PM 8891224 ER PT J AU Molldrem, J Dermime, S Parker, K Jiang, YZ Mavroudis, D Hensel, N Fukushima, P Barrett, AJ AF Molldrem, J Dermime, S Parker, K Jiang, YZ Mavroudis, D Hensel, N Fukushima, P Barrett, AJ TI Targeted T-cell therapy for human leukemia: Cytotoxic T lymphocytes specific for a peptide derived from proteinase 3 preferentially lyse human myeloid leukemia cells SO BLOOD LA English DT Article ID TUMOR-ANTIGENS; BONE-MARROW; RECOGNITION; CANCER; MYELOBLASTIN; IMMUNITY; REGION; SELF AB Proteinase 3 is present in high concentration in the primary granules of acute and chronic myeloid leukemia blasts, and may represent a potential T-cell target antigen. We screened proteinase 3 against the binding motif of HLA-A2.1. Based on its high predicted binding, a 9-mer peptide, ''PR-1,'' was synthesized and tested for binding to HLA-A2.1 using the T2 cell line. PR-1 at 100 mu g/mL significantly increased expression of HLA-A2.1, with median channel of fluorescence increasing from 22 to 294. Binding half-life was determined to be 1,460 minutes by I-125-labeled beta(2) . microglobulin incorporation. HLA-A2.1(+) peripheral blood mononuclear cells from a normal donor were used to generate a T-cell line specific for PR-1. The line demonstrated 85% PR-1-specific lysis at an E:T ratio of 50:1, compared with 20% lysis without PR-1, using T2 cells as targets. It also showed 79% specific lysis to fresh chronic myelogenous leukemia blasts, 54% to fresh acute myelogenous leukemia blasts, and only background lysis (<20%) to HLA-A2.1(+) normal allogeneic marrow cells. The amount of lysis of HLA-A2.1(+) myeloid cells was proportional to cytoplasmic proteinase 3 expression. Thus, HLA-A2.1-restricted cytotoxic T cells, raised against a peptide contained in proteinase 3, preferentially lysed fresh human leukemic cells. This is a US government work, There are no restrictions on its use. C1 NIAID,MOL STRUCT LAB,NIH,BETHESDA,MD 20892. RP Molldrem, J (reprint author), NHLBI,HEMATOL BRANCH,BONE MARROW TRANSPLANT UNIT,NIH,BLDG 10,ROOM 7C103,9000 ROCKVILLE PIKE,BETHESDA,MD 20892, USA. RI Dermime, Said/C-6235-2009; OI Dermime, Said/0000-0002-5526-7496; Parker, Kenneth/0000-0002-6282-2478 NR 40 TC 254 Z9 259 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0006-4971 J9 BLOOD JI Blood PD OCT 1 PY 1996 VL 88 IS 7 BP 2450 EP 2457 PG 8 WC Hematology SC Hematology GA VK747 UT WOS:A1996VK74700008 PM 8839835 ER PT J AU Preas, HL Reda, D Tropea, M Vandivier, RW Banks, SM Agosti, JM Suffredini, AF AF Preas, HL Reda, D Tropea, M Vandivier, RW Banks, SM Agosti, JM Suffredini, AF TI Effects of recombinant soluble type I interleukin-1 receptor on human inflammatory responses to endotoxin SO BLOOD LA English DT Article ID TUMOR-NECROSIS-FACTOR; IL-1 RECEPTOR; INTRAVENOUS ENDOTOXIN; CYTOKINE SYNTHESIS; SEPSIS SYNDROME; HUMAN MONOCYTES; DECOY RECEPTOR; SEPTIC SHOCK; PHASE-I; ANTAGONIST AB Effects of soluble recombinant human type I interleukin-l receptor (sIL-1R(1)) were evaluated in 18 volunteers given intravenous endotoxin and randomized to placebo (n = 6), low-dose (n = 6), or high-dose (n = 6) sIL-1R(1). Soluble IL-1R(1) decreased IL-1 beta (P = .001), but decreased IL-1ra (P = .0001), and resulted in 10-fold and 43-fold dose-related increases in sIL-1R(1)-IL-1ra complexes compared with placebo (P less than or equal to .001). High-dose sIL-1R(1) was associated with increased levels of immunoactive tumor necrosis factor-alpha (P = .02), IL-8 (P = .0001), and cell-associated IL-1 beta (P = .047). C-reactive protein levels were higher after sIL-1R(1) than placebo (P = .035), Soluble IL-1R(1) decreased the severity of chills (P = .03), but did not alter other symptoms, changes in temperature, systemic hemodynamic responses, or changes in leukocyte and platelet number. Thus, sIL-1R(1) had no discernable antiinflammatory effect following endotoxin administration due in part to low levels of circulating IL-1 beta and neutralization of IL-1ra inhibitory function. This latter interaction represents an indirect mechanism of agonist activity elicited by sIL-1R(1), and may contribute to increases in inflammatory mediators, limiting therapy with sIL-1R(1) during endotoxemia. This is a US government work. There are no restrictions on its use. C1 IMMUNEX CORP,SEATTLE,WA. RP Preas, HL (reprint author), NIH,DEPT CRIT CARE MED,WARREN G MAGNUSON CLIN CTR,BLDG 10,ROOM 7D-43,BETHESDA,MD 20892, USA. NR 55 TC 42 Z9 44 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0006-4971 J9 BLOOD JI Blood PD OCT 1 PY 1996 VL 88 IS 7 BP 2465 EP 2472 PG 8 WC Hematology SC Hematology GA VK747 UT WOS:A1996VK74700010 PM 8839837 ER PT J AU Beran, M Kantarjian, H OBrien, S Koller, C AlBitar, M Arbuck, S Pierce, S Moore, M Abbruzzese, JL Andreeff, M Keating, M Estey, E AF Beran, M Kantarjian, H OBrien, S Koller, C AlBitar, M Arbuck, S Pierce, S Moore, M Abbruzzese, JL Andreeff, M Keating, M Estey, E TI Topotecan, a topoisomerase I inhibitor, is active in the treatment of myelodysplastic syndrome and chronic myelomonocytic leukemia SO BLOOD LA English DT Article ID CHRONIC MYELOGENOUS LEUKEMIA; DOSE CYTOSINE-ARABINOSIDE; REVERSE DOT-BLOT; PHASE-II; MYELOID LEUKEMIAS; MUTATIONS; RAS; 5-AZACYTIDINE; CHEMOTHERAPY; CAMPTOTHECIN AB The aim of this study was to evaluate the activity of topotecan in patients with myelodysplastic syndrome (MDS) and chronic myelomonocytic leukemia (CMML). Forty-seven patients with a diagnosis of MDS (n = 22) or CMML (n = 25) were treated. The median age was 66 years. Chromosomal abnormalities were present in 70% and thrombocytopenia less than 50 x 10(3)/mu l in 51%. Evaluation of outcome and of differences among subgroups was performed according to standard methods; the criteria for response were those used for acute leukemia, Topotecan was administered as 2 mg/m(2) by continuous infusion over 24 hours daily for 5 days (10 mg/m(2) per course) every 3 to 4 weeks until remission, then once every month for a maximum of 12 courses. Thirteen patients (28%) achieved a complete response (CR) and six (13%) had hematologic improvement, A CR was achieved in six of 22 patients with MDS (27%) and in seven of 25 with CMML (28%). All eight patients who presented with cytogenetic abnormalities (five chromosome 5 or 7 abnormalities) who achieved CR were cytogenetically normal in CR. Characteristics for which there was evidence of association with a higher response rate were lack of prior chemotherapy, less than 10% marrow monocytes, and absence of RAS oncogene mutations. In contrast, CR rates were similar in patients with or without abnormal karyotypes. Mucositis occurred in 64% of patients (severe in 19%) and diarrhea in 32% (severe in 13%). Febrile episodes occurred in 85% of patients and documented infections in 47%. With a median follow-up duration of 8 months, the 12-month survival rate was 38%, median survival time 10.5 months, and median remission duration 7.5 months. We conclude that topotecan has significant activity in MDS and CMML, with acceptable side effects. Future studies will investigate topotecan combined with topoisomerase II reactive agents, cytarabine, or hypomethylating agents (azacytidine and decitabine). (C) 1996 by The American Society of Hematology. C1 NCI, BETHESDA, MD 20892 USA. RP UNIV TEXAS, MD ANDERSON CANC CTR, DEPT HEMATOL, LEUKEMIA SECT, BOX 61, HOUSTON, TX 77030 USA. FU NCI NIH HHS [CA 56887-01] NR 42 TC 95 Z9 98 U1 0 U2 0 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 2021 L ST NW, SUITE 900, WASHINGTON, DC 20036 USA SN 0006-4971 EI 1528-0020 J9 BLOOD JI Blood PD OCT 1 PY 1996 VL 88 IS 7 BP 2473 EP 2479 PG 7 WC Hematology SC Hematology GA VK747 UT WOS:A1996VK74700011 PM 8839838 ER PT J AU Zhan, SX Vazquez, N Zhan, SL Wientjes, FB Budarf, ML Schrock, E Ried, T Green, ED Chanock, SJ AF Zhan, SX Vazquez, N Zhan, SL Wientjes, FB Budarf, ML Schrock, E Ried, T Green, ED Chanock, SJ TI Genomic structure, chromosomal localization, start of transcription, and tissue expression of the human p40-phox, a new component of the nicotinamide adenine dinucleotide phosphate-oxidase complex SO BLOOD LA English DT Article ID CHRONIC GRANULOMATOUS-DISEASE; RESPIRATORY BURST OXIDASE; PHAGOCYTE NADPH OXIDASE; LEUKEMIA-CELL-LINE; NEUTROPHIL CYTOCHROME-B; 2 CYTOSOLIC COMPONENTS; SRC HOMOLOGY-3 DOMAINS; PHILADELPHIA-CHROMOSOME; 4 COMPONENTS; LIGHT CHAIN AB p40-phox is a newly isolated cytosolic component of the nicotinamide adenine dinucleotide phosphate (NADPH)-oxidase that copurifies with p87-phox, Although its function is not well defined, preliminary evidence indicates that it is a component of the cytosolic complex. We report the characterization of the human p40-phox gene, which is single copy and spans approximately 18 kb with 10 exons. Based on fluorescent in situ hybridization (FISH) studies and analysis of somatic hybrid cell lines, the chromosomal location of p40-phox is human chromosome 22q13.1. The start of transcription has been mapped to bp -156. The expression of p40-phox message is restricted to hematopoietic cells. In addition to identifying the mRNA transcript on Northern blot analysis in cells known to express components of the NADPH-oxidase, polymorphonuclear leukocytes, monocytes, B lymphoblastoid cell lines, and eosinophils, p40-phox is also expressed in two other cell types of white cell lineage, mast cells, and basophils. In addition, the mRNA for p40-phox is expressed in megakaryocytic cells, but not in erythroid cells. (C) 1996 by The American Society of Hematology. C1 NCI, PEDIAT BRANCH, NATL CTR HUMAN GENOME RES, NIH, BETHESDA, MD 20892 USA. UCL, DIV MOL MED, LONDON, ENGLAND. CHILDRENS HOSP PHILADELPHIA, DIV HUMAN GENET & MOL BIOL, PHILADELPHIA, PA 19104 USA. FU NHGRI NIH HHS [HG00425] NR 57 TC 54 Z9 56 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 OCT 1 PY 1996 VL 88 IS 7 BP 2714 EP 2721 PG 8 WC Hematology SC Hematology GA VK747 UT WOS:A1996VK74700040 PM 8839867 ER PT J AU Kerfoot, C Wienecke, R Menchine, M Emelin, J Maize, JC Welsh, CT Norman, MG DeClue, JE Vinters, HV AF Kerfoot, C Wienecke, R Menchine, M Emelin, J Maize, JC Welsh, CT Norman, MG DeClue, JE Vinters, HV TI Localization of tuberous sclerosis 2 mRNA and its protein product tuberin in normal human brain and in cerebral lesions of patients with tuberous sclerosis SO BRAIN PATHOLOGY LA English DT Article ID EKER RAT; GENE; IDENTIFICATION; EPILEPSY; TISSUES; REGION; TSC2 AB Tuberous sclerosis (TSC), an autosomal dominant disorder, is characterized by malformations, hamartomas and tumors in various organs including the brain. TSC is genetically linked to two loci: TSC1 on chromosome 9q34 and TSC2 on 16p13.3. TSC2 has been cloned, sequenced and encodes a protein (tuberin) which functions as a tumor suppressor. We have analyzed the distribution of TSC2 mRNA and tuberin in the brains of TSC patients and non-affected individuals using both autopsy and biopsy material. High levels of transcript and protein expression were observed in choroid plexus epithelium, ependymal cells, most brainstem and spinal cord motor neurons, Purkinje cells and the external granule cell layer of the cerebellum in both TSC and control cases. Individual balloon cells from TSC patients showed very faint expression while other glia showed no expression of either transcript or tuberin. Neocortical and hippocampal neurons expressed high levels of TSC2 transcript, but only modest levels of tuberin. The internal granule cell layer of the cerebellum expressed abundant transcript but low levels of tuberin. These observations suggest either that tuberin expression is controlled at the level of both transcription and translation or the antibody and in-situ hybridization recognize different splice variants of the TSC2 gene. In TSC patients, dysmorphic cytomegalic neurons expressed high levels of tuberin and transcript, particularly when in an 'ectopic' location. individual cells within subependymal giant cell astrocytomas (SEGAs) and hamartomas from TSC patients expressed moderate to high levels of TSC2 transcript and tuberin. While the TSC2 transcript is widely expressed primarily within neurons, tuberin is demonstrable primarily within dysplastic/cytomegalic cells of the cortex and subependymal hamartomas/SEGAs. CNS expression of tuberin is unique in that primarily non-dividing cells express it in this location, whereas extra-CNS expression of tuberin is mainly found in actively proliferating cell types such as epithelium. C1 NCI,CELLULAR ONCOL LAB,BETHESDA,MD 20892. BRITISH COLUMBIA CHILDRENS HOSP,DEPT PATHOL & LAB MED,VANCOUVER,BC V6H 3V4,CANADA. UNIV BRITISH COLUMBIA,VANCOUVER,BC V6H 3V4,CANADA. UNIV CALIF LOS ANGELES,INST BRAIN RES,LOS ANGELES,CA 90095. UNIV CALIF LOS ANGELES,MENTAL RETARDAT RES CTR,LOS ANGELES,CA 90095. RP Kerfoot, C (reprint author), UNIV CALIF LOS ANGELES,SCH MED,DEPT PATHOL & LAB MED,HLTH SCI CTR,ROOM 13-388,LOS ANGELES,CA 90095, USA. FU NINDS NIH HHS [P01 NS 28383] NR 31 TC 50 Z9 51 U1 0 U2 2 PU INT SOC NEUROPATHOLOGY PI ZURICH PA ISN JOURNAL PO BOX, CH-8033 ZURICH, SWITZERLAND SN 1015-6305 J9 BRAIN PATHOL JI Brain Pathol. PD OCT PY 1996 VL 6 IS 4 BP 367 EP 375 DI 10.1111/j.1750-3639.1996.tb00866.x PG 9 WC Clinical Neurology; Neurosciences; Pathology SC Neurosciences & Neurology; Pathology GA VQ897 UT WOS:A1996VQ89700001 PM 8944308 ER PT J AU Campbell, EM Watson, ML Proudfoot, AEI Wells, TNC Yoshimura, T Westwick, J AF Campbell, EM Watson, ML Proudfoot, AEI Wells, TNC Yoshimura, T Westwick, J TI Guinea-pig RANTES activates human, but not guinea-pig, eosinophils SO BRITISH JOURNAL OF PHARMACOLOGY LA English DT Meeting Abstract ID CYTOKINE RANTES C1 UNIV BATH,DEPT PHARMACOL,BATH BA2 7AY,AVON,ENGLAND. GLAXO INST MOL BIOL SA,CH-1211 GENEVA,SWITZERLAND. NCI,FCRDC,BETHESDA,MD 20892. NR 3 TC 0 Z9 0 U1 0 U2 0 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE, HAMPSHIRE, ENGLAND RG21 6XS SN 0007-1188 J9 BRIT J PHARMACOL JI Br. J. Pharmacol. PD OCT PY 1996 VL 119 SU S BP P50 EP P50 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA VK188 UT WOS:A1996VK18800050 ER PT J AU White, AM Westwick, J Smith, AW Yoshimura, T Watson, ML AF White, AM Westwick, J Smith, AW Yoshimura, T Watson, ML TI Guinea-pig tumour necrosis factor-induced airway inflammation: Inhibition by interleukin-13 SO BRITISH JOURNAL OF PHARMACOLOGY LA English DT Meeting Abstract C1 UNIV BATH,DEPT PHARMACOL,BATH BA2 7AY,AVON,ENGLAND. NCI,FREDERICK CANC RES & DEV CTR,FREDERICK,MD. NR 2 TC 0 Z9 0 U1 0 U2 1 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE, HAMPSHIRE, ENGLAND RG21 6XS SN 0007-1188 J9 BRIT J PHARMACOL JI Br. J. Pharmacol. PD OCT PY 1996 VL 119 SU S BP P47 EP P47 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA VK188 UT WOS:A1996VK18800047 ER PT J AU Garraud, O Nutman, TB AF Garraud, O Nutman, TB TI The role of cytokines in human B-cell differentiation into immunoglobulin-secreting cells SO BULLETIN DE L INSTITUT PASTEUR LA English DT Review DE B lymphocyte; cytokine; immunoglobulin; humans; differentiation; isotype switching; antibody production; review ID HUMAN LYMPHOCYTES-B; BLOOD MONONUCLEAR-CELLS; GROWTH-FACTOR-BETA; CD4+ T-CELLS; NECROSIS-FACTOR-ALPHA; HUMAN IGE SYNTHESIS; HUMAN-BONE-MARROW; ANTI-CD40 MONOCLONAL-ANTIBODIES; CD40-CD40 LIGAND INTERACTION; IL-13 INDUCES PROLIFERATION C1 NIAID,PARASIT DIS LAB,NIH,BETHESDA,MD 20892. RP Garraud, O (reprint author), INST PASTEUR,UNITE IMMUNOL,BP 220,DAKAR,SENEGAL. NR 236 TC 28 Z9 28 U1 0 U2 2 PU EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER PI PARIS PA 141 RUE JAVEL, 75747 PARIS, FRANCE SN 0020-2452 J9 B I PASTEUR JI Bull. Inst. Pasteur PD OCT-DEC PY 1996 VL 94 IS 4 BP 285 EP 309 DI 10.1016/S0020-2452(97)87084-4 PG 25 WC Immunology; Microbiology; Virology SC Immunology; Microbiology; Virology GA WL362 UT WOS:A1996WL36200002 ER PT J AU Johnson, FE AF Johnson, FE TI NLM planning grants for the education and training of health sciences librarians - Introduction SO BULLETIN OF THE MEDICAL LIBRARY ASSOCIATION LA English DT Editorial Material RP Johnson, FE (reprint author), NATL LIB MED,DIV EXRAMURAL PROGRAMS,8600 ROCKVILLE PIKE,BETHESDA,MD 20894, USA. NR 2 TC 1 Z9 1 U1 0 U2 0 PU MED LIBRARY ASSN PI CHICAGO PA SUITE 300 6 N MICHIGAN AVE, CHICAGO, IL 60602 SN 0025-7338 J9 B MED LIBR ASSOC JI Bull. Med. Libr. Assoc. PD OCT PY 1996 VL 84 IS 4 BP 514 EP 514 PG 1 WC Information Science & Library Science SC Information Science & Library Science GA VQ510 UT WOS:A1996VQ51000008 ER PT J AU Kuznetsov, S Robey, PG AF Kuznetsov, S Robey, PG TI Species differences in growth requirements for bone marrow stromal fibroblast colony formation in vitro SO CALCIFIED TISSUE INTERNATIONAL LA English DT Article DE marrow; stromal; fibroblast; colony; formation ID CELLS CFU-F; FORMING CELLS; HEMATOPOIETIC MICROENVIRONMENT; DIFFERENTIATION; CULTURES; INVITRO; PROLIFERATION; STIMULATION; EXPRESSION; PROGENY AB The marrow stromal fibroblast (MSF) population has been shown to include precursor cells for at least five types of connective tissue: bone, cartilage, adipose tissue, fibrous tissue, and hematopoiesis-supporting reticular stroma. In this study, growth requirements for MSF colony formation were studied in vitro. In order to exclude the influence of nonadherent cells, after a period of initial adhesion of bone marrow cells in serum-containing medium nonadherent cells were removed. Further cultivation was carried out in either serum-containing or serum-free conditions, with or without feeder cells (irradiated bone marrow cells). This approach revealed differences between animal species in initial MSF growth requirements. In serum-containing conditions, mouse MSF precursor cells (colony-forming units-fibroblast, CFU-Fs) were shown to be feeder cell dependent: MSF colonies were formed only in the pres ence of feeder cells. Guinea pig CFU-Fs were partially feeder cell dependent, whereas human CFU-Fs were feeder cell independent. In serum-free conditions, CFU-Fs of all three species were feeder cell dependent. The difference between the growth requirements for mouse and human MSFs was not caused by serum origin or concentration, feeder cell origin, or differences in the preparation of marrow cell suspensions. RP Kuznetsov, S (reprint author), NIDR,BONE RES BRANCH,NIH,BETHESDA,MD 20892, USA. RI Robey, Pamela/H-1429-2011 OI Robey, Pamela/0000-0002-5316-5576 NR 37 TC 66 Z9 68 U1 0 U2 0 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 SN 0171-967X J9 CALCIFIED TISSUE INT JI Calcif. Tissue Int. PD OCT PY 1996 VL 59 IS 4 BP 265 EP 270 DI 10.1007/s002239900121 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VJ110 UT WOS:A1996VJ11000009 PM 8781050 ER PT J AU Gail, MH Fears, TR Hoover, RN Chandler, DW Donaldson, JL Hyer, MB Pee, D Ricker, WV Siiteri, PK Stanczyk, FZ Vaught, JB Ziegler, RG AF Gail, MH Fears, TR Hoover, RN Chandler, DW Donaldson, JL Hyer, MB Pee, D Ricker, WV Siiteri, PK Stanczyk, FZ Vaught, JB Ziegler, RG TI Reproducibility studies and interlaboratory concordance for assays of serum hormone levels: Estrone, estradiol, estrone sulfate, and progesterone SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Article ID POSTMENOPAUSAL WOMEN; PLASMA PROGESTERONE; SEX-HORMONES; RADIOIMMUNOASSAY; RELIABILITY AB We conducted studies to measure sources of assay variability for estrone, estradiol, estrone sulfate, and progesterone for postmenopausal women (It = 5) and for women in the mid-follicular (n = 5) and mid-luteal (n = 5) phases of the menstrual cycle, A single blood sample from each woman was divided into 2.5-ml aliquots and stored at -70 degrees C, and sets of two aliquots were sent at monthly intervals to each of three laboratories (four for progesterone). Each aliquot was analyzed in duplicate, Thus, within each menstrual category, we were able to estimate the components of variance due to variation among women, variation among aliquots, variation among duplicate measurements, and variation among the 4 analysis days, Using the logarithm of assay measurements, we estimated the percentage of variance attributable to variation among women in each menstrual category, 100 <(rho)over tilde>, where <(rho)over tilde> is the estimated intraclass correlation, For each assay, 100 <(rho)over tilde> exceeded 90% for midfollicular and mid-luteal women, For postmenopausal women, values of 100 <(rho)over tilde> exceed 84% for estrone in two laboratories, Values of 100 <(rho)over tilde> were lower for progesterone in postmenopausal women, although a value of 84% was estimated from one laboratory, These studies indicate that estrogen assays over a period of 3 months permit reliable comparisons among women in a given menstrual category, Progesterone measurements are likewise reliable for women in the mid-follicular and mid-luteal phases but somewhat less satisfactory for postmenopausal women, These assessments of variability pertain only to laboratory techniques and do not allow for secular variation in intra-woman hormone levels, Moreover, although these measurements tend to be reliable enough for making comparisons among women, estimates of coefficients of variation for estrogens are about 10% for mid-follicular and mid-luteal phase women and about 11-20% for postmenopausal women, Coefficients of variation for progesterone are about 10% for mid-luteal, 20% for mid-follicular, and 30% for postmenopausal women. C1 NCI,NIH,ENVIRONM EPIDEMIOL BRANCH,BETHESDA,MD 20892. ENDOCRINE SCI,CALABASAS,CA 91304. INFORMAT MANAGEMENT SERV INC,ROCKVILLE,MD 20852. WOMENS & CHILDRENS HOSP,LOS ANGELES,CA 90033. MICROBIOL ASSOCIATES INC,ROCKVILLE,MD 20850. RP Gail, MH (reprint author), NCI,NIH,BIOSTAT BRANCH,EXECUT PLAZA N,ROOM 431,MSC 7368,6130 EXECUTIVE B,BETHESDA,MD 20892, USA. NR 22 TC 27 Z9 31 U1 0 U2 6 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 1055-9965 J9 CANCER EPIDEM BIOMAR JI Cancer Epidemiol. Biomarkers Prev. PD OCT PY 1996 VL 5 IS 10 BP 835 EP 844 PG 10 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA VM565 UT WOS:A1996VM56500010 PM 8896895 ER PT J AU Wang, TTY Phang, JM AF Wang, TTY Phang, JM TI Effect of N-(4-hydroxyphenyl)retinamide on apoptosis in human breast cancer cells SO CANCER LETTERS LA English DT Article DE apoptosis; Bax; Bcl-2; breast cancer; estradiol; retinoids ID RETINOIC ACID; BCL-2 PROTEIN; ESTROGEN; EXPRESSION; TISSUE; DEATH; CYCLE AB The synthetic retinoid N-(4-hydroxyphenyl)retinamide (4-HPR) has been used in breast cancer prevention and treatment. However, the molecular mechanisms mediating the effects of 4-HPR remain elusive. In the present study, we examined the effects of 4-HPR on components of apoptosis pathway (i.e. Bcl-2 and Bax) and apoptotic death in both estrogen receptor-positive and estrogen receptor-negative cell lines. We found that: (1) 4-HPR treatment resulted in decreased Bcl-2 mRNA but not Bax mRNA levels; (2) the effect of 4-HPR on Bcl-2 mRNA level was different from other retinoids; (3) 4-HPR treatment induced apoptosis in both estrogen receptor-positive and -negative cells. Hence, the breast cancer chemopreventive properties of 4-HPR may involve modulation of apoptosis pathways. RP Wang, TTY (reprint author), NCI, FREDERICK CANC RES & DEV CTR, LAB NUTR & MOL REGULAT,NIH, BLDG 434-2, POB B, FREDERICK, MD 21702 USA. NR 19 TC 39 Z9 39 U1 0 U2 1 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0304-3835 J9 CANCER LETT JI Cancer Lett. PD OCT 1 PY 1996 VL 107 IS 1 BP 65 EP 71 DI 10.1016/0304-3835(96)04344-3 PG 7 WC Oncology SC Oncology GA VP941 UT WOS:A1996VP94100010 PM 8913268 ER PT J AU Kuehl, WM Brents, LA Chesi, M Bergsagel, PL AF Kuehl, WM Brents, LA Chesi, M Bergsagel, PL TI Selective expression of one c-myc allele in two human myeloma cell lines SO CANCER RESEARCH LA English DT Article ID BURKITTS-LYMPHOMA; MULTIPLE-MYELOMA; CODING REGION; MUTATIONS; TRANSLOCATIONS; PROTOONCOGENE; VARIANT; MICE AB Chromosomal translocations or DNA rearrangements affecting c-myc occur in almost all murine plasmacytoma and human Burkitt's lymphoma tumors and are associated with a high incidence of exon 2 missense mutations and selective expression of the affected allele. Screening nine multiple myeloma cell lines, we identified no exon 2 missense mutations but did identify two lines with single, silent mutations in exon 1 and exon 2, respectively. Each of these informative multiple myeloma cell lines selectively expresses only one c-myc allele despite the apparent absence of chromosomal translocations or DNA rearrangements affecting c-myc. C1 NCI,NAVY MED ONCOL BRANCH,NIH,BETHESDA,MD 20889. CORNELL UNIV,COLL MED,DIV HEMATOL ONCOL,NEW YORK,NY 10021. RI Bergsagel, Peter/A-7842-2011 OI Bergsagel, Peter/0000-0003-1523-7388 NR 19 TC 8 Z9 8 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 0008-5472 J9 CANCER RES JI Cancer Res. PD OCT 1 PY 1996 VL 56 IS 19 BP 4370 EP 4373 PG 4 WC Oncology SC Oncology GA VK307 UT WOS:A1996VK30700019 PM 8813127 ER PT J AU Dong, JT Suzuki, H Pin, SS Bova, GS Schalken, JA Isaacs, WB Barrett, JC Isaacs, JT AF Dong, JT Suzuki, H Pin, SS Bova, GS Schalken, JA Isaacs, WB Barrett, JC Isaacs, JT TI Down-regulation of the KAI1 metastasis suppressor gene during the progression of human prostatic cancer infrequently involves gene mutation or allelic loss SO CANCER RESEARCH LA English DT Article ID SYNCYTIUM FORMATION; POLYMORPHISMS; DNA AB The KAI1 gene, located on human chromosome 11p11.2, suppresses tumor metastasis when expressed in certain cancer cells. To evaluate whether dysregulation of KAI1 occurs during the progression of human prostatic cancer, protein expression, mutation, and allelic loss of KAI1 were analyzed using a tissue bank of 98 primary cancers and 32 metastases. By immunohistochemical staining, high levels of KAI1 protein are detected in the epithelial but not stromal compartment of normal prostatic and benign prostatic hyperplasia tissue. In epithelial cells, KAI1 protein is expressed on the plasma membrane. KAI1 protein expression is downregulated in more than 70% of the 49 primary prostatic cancers from untreated patients. In 10 such untreated patients, down-regulation of KAI1 protein occurred in all of the lymph node metastases examined. In 15 patients with metastatic disease who had failed androgen ablation therapy, more than 90% of the primary prostatic cancers had downregulation, with 60% having no KAI1 protein expression. Primers derived from the sequences flanking each exon of KAI1 were used to analyze KAI1 mutation and allelic loss by the method of PCR-single-strand conformational polymorphism. Using this method, no point mutation or allelic loss was detected in metastases from 10 patients. No allelic loss was detected in an additional 34 primary and 12 lymph node metastases via microsatellite analysis using the marker D11S1344, which is located in the region of KAI1. These results demonstrate that KAI1 protein expression is consistently down-regulated during the progression of human prostatic cancer and that this down-regulation does not commonly involve either mutation or allelic loss of the KAI1 gene. C1 JOHNS HOPKINS UNIV,SCH MED,CTR ONCOL,BALTIMORE,MD 21231. JOHNS HOPKINS UNIV,SCH MED,DEPT UROL,BALTIMORE,MD 21231. JOHNS HOPKINS UNIV,SCH MED,JAMES BUCHANAN BRADY UROL INST,BALTIMORE,MD 21231. JOHNS HOPKINS UNIV,SCH MED,DEPT PATHOL,BALTIMORE,MD 21231. UNIV NIJMEGEN,UROL RES LAB,NIJMEGEN,NETHERLANDS. NIEHS,RES TRIANGLE PK,NC 27709. RI Schalken, Jack/B-1277-2014 OI Schalken, Jack/0000-0001-8274-7797 FU NCI NIH HHS [CA 58236] NR 16 TC 231 Z9 260 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 0008-5472 J9 CANCER RES JI Cancer Res. PD OCT 1 PY 1996 VL 56 IS 19 BP 4387 EP 4390 PG 4 WC Oncology SC Oncology GA VK307 UT WOS:A1996VK30700023 PM 8813131 ER PT J AU Bai, RL Schwartz, RE Kepler, JA Pettit, GR Hamel, E AF Bai, RL Schwartz, RE Kepler, JA Pettit, GR Hamel, E TI Characterization of the interaction of cryptophycin 1 with tubulin: Binding in the Vinca domain, competitive inhibition of dolastatin 10 binding, and an unusual aggregation reaction SO CANCER RESEARCH LA English DT Article ID ANTINEOPLASTIC AGENTS; GAMMA-TUBULIN; NATURAL-PRODUCTS; HALICHONDRIN-B; PHOMOPSIN-A; COMPONENT; POLYMERIZATION; CYTOTOXICITY; MICROTUBULES; VINBLASTINE AB The antimitotic depsipeptide cryptophgcin 1 (CP1) was compared to the antimitotic peptide dolastatin 10 (D10) as an antiproliferative agent and in its interactions with purified tubulin. The potent activity of CP1 as an inhibitor of cell growth was confirmed. The agent had an IC50 of 20 pM against L1210 murine leukemia cells versus 0.5 nM for D10. Both drugs were comparable as inhibitors of the glutamate-induced assembly of purified tubulin, with D10 being slightly more potent. CP1, like D10, was a noncompetitive inhibitor of the binding of [H-3]vinblastine to tubulin (apparent K-i, 3.9 mu M); and the depsipeptide was a competitive inhibitor of the binding of [H-3]D10 to tubulin (apparent K-i, 2.1 mu M). CP1 was less potent than D10 as an inhibitor of nucleotide exchange on tubulin, but the two drugs were equivalent in stabilizing the colchicine binding activity of tubulin, CP1, like D10, caused the formation of extensive structured aggregates of tubulin when present in stoichiometric amounts relative to the protein. Whereas at lower concentrations the drugs were equivalent in causing formation of small oligomers detected by gel permeation highperformance liquid chromatography, there were notable differences in the aggregation reactions induced by the two drugs. The electron micrographic appearance of the D10-induced aggregate differed substantially from that of the CP1 induced aggregate. With D10, but not CP1, aggregate morphology was greatly altered in the presence of microtubule-associated proteins. Finally, although CP1 caused the formation of massive aggregates, as did D10, there was little turbidity change with the depsipeptide as opposed to the peptide. C1 NCI,MOL PHARMACOL LAB,DIV BASIC SCI,NIH,BETHESDA,MD 20892. MERCK & CO INC,MERCK SHARP & DOHME RES LABS,RAHWAY,NJ 07065. RES TRIANGLE INST,RES TRIANGLE PK,NC 27709. ARIZONA STATE UNIV,CANC RES INST,TEMPE,AZ 85287. ARIZONA STATE UNIV,DEPT CHEM,TEMPE,AZ 85287. NR 37 TC 75 Z9 78 U1 0 U2 2 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 0008-5472 J9 CANCER RES JI Cancer Res. PD OCT 1 PY 1996 VL 56 IS 19 BP 4398 EP 4406 PG 9 WC Oncology SC Oncology GA VK307 UT WOS:A1996VK30700025 PM 8813133 ER PT J AU Goldwasser, F Shimizu, T Jackman, J Hoki, Y OConnor, PM Kohn, KW Pommier, Y AF Goldwasser, F Shimizu, T Jackman, J Hoki, Y OConnor, PM Kohn, KW Pommier, Y TI Correlations between S and G(2) arrest and the cytotoxicity of camptothecin in human colon carcinoma cells SO CANCER RESEARCH LA English DT Article ID DNA TOPOISOMERASE-I; MAMMALIAN-CELLS; STRAND BREAKS; SACCHAROMYCES-CEREVISIAE; ENHANCED SENSITIVITY; CLEAVABLE COMPLEXES; REPLICATION FORKS; CROSS-LINKING; CYCLE ARREST; LUNG-CANCER AB Previous cell line comparisons indicated that neither S-phase fraction nor topoisomerase I (top1) levels are sufficient to predict camptothecin (CPT) cytotoxicity (F. Goldwasser et al., Cancer Res., 55: 2116-2121, 1995.). To identify new determinants for CPT activity, two mutant p53 human colon cancer cell lines, SW620 and KM12, that were previously reported to have similar top1 levels and differential sensitivity to CPT were studied. No difference in the kinetics of top1-mediated DNA single-strand breaks or DNA synthesis inhibition were observed after 1 h exposure to 1 mu M CPT. Pulse-labeling alkaline elution show;ed deficiency of damaged replicon elongation in the more sensitive SW620 cells. Consistently, flow cytometry analyses showed that KM12 was arrested in G(2), whereas SW620 cells were irreversibly blocked in S phase. Aphidicolin protection was minimal in KM12 and more pronounced in the more sensitive SW620 cells. Thus, CPT appears to have two cytotoxic mechanisms, one protectable by aphidicolin and present in SW620 and the other not protectable by aphidicolin and common to both cell fines. SW620 exhibited also a greater capacity to break through the G(2) checkpoint after DNA damage. Consistently, SW620 cells failed to down-regulate cyclin B-cdc2 kinase activity, whereas KM12 cells down-regulated cyclin B/cdc2 kinase activity within 30 min to 20% of control level after CPT treatment, Analysis of the 7 human colon carcinoma cell lines of the NCI Anticancer Drug Screen showed that defects in replicon elongation and G(2) breakthrough capability correlate with sensitivity to CPT. Our results suggest that misrepair of damaged replicons and/or alterations in DNA damage checkpoints is critical to defining chemosensitivity to CPT-induced top1-cleavable complexes and that CPT appears to hal e tno cytotoxic mechanisms, one protectable by aphidicolin, and the other not. C1 NCI,NIH,MOL PHARMACOL LAB,DIV BASIC SCI,BETHESDA,MD 20892. NR 55 TC 141 Z9 151 U1 1 U2 2 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 0008-5472 J9 CANCER RES JI Cancer Res. PD OCT 1 PY 1996 VL 56 IS 19 BP 4430 EP 4437 PG 8 WC Oncology SC Oncology GA VK307 UT WOS:A1996VK30700029 PM 8813137 ER PT J AU Li, MF Muller, J Xu, F Hearing, VJ Gorelik, E AF Li, MF Muller, J Xu, F Hearing, VJ Gorelik, E TI Inhibition of melanoma-associated antigen expression and ecotropic retrovirus production in B16BL6 melanoma cells transfected with major histocompatibility complex class I genes SO CANCER RESEARCH LA English DT Article ID MURINE LEUKEMIA-VIRUS; TNF-MEDIATED CYTOTOXICITY; B-16 MELANOMA; MONOCLONAL-ANTIBODIES; INCREASED SENSITIVITY; H-2K(B) GENE; H-2 GENES; DNA; ORGANIZATION; METASTASES AB We have previously reported that expression of the melanoma-associated antigen (MAA) recognized by MM2-9B6 monoclonal antibody in B16 melanoma was closely associated with C-type ecotropic retroviral particle production. Our present data show that this MAA is encoded by the env gene of an ecotropic retrovirus produced by B16 melanoma cells. Transfection of H-2K(b) or H-2K(d) genes into two individual clones isolated from B16BL6 melanoma, BL6-8 (H-2K(b)-, H-2D(b)+) and BL6-2- (H-2K(b)-, H-2D(b)-), resulted in a loss of MAA expression. Electron immunohistochemical analysis of melanoma cells and reverse transcriptase assay revealed that the loss of MAA expression in the H-2K gene-transfected cells paralleled the elimination of retroviral particles. In contrast, expression of the endogenous H-2D(b) gene or transfection with the H-2D(d) or H-2L(d) gene had no effect on MAA expression or retrovirus production. Northern blot analysis showed equivalent retroviral messages in retrovirus-producing and -nonproducing BL6 melanoma clones. Southern blot analysis revealed that H-2K(b)-negative BL6 melanoma cells contain at least four different ecotropic retroviruses with different insertion sites that somatically emerged during malignant transformation or progression. Restriction enzyme analysis showed various changes in proviral DNAs from the H-2K(b)- and H-2K(d)-transfected cells that failed to produce retroviral particles. The observed alterations in the patterns of PstI- and HindIII-digested proviral DNA mere found to be due to the appearance of PstI and loss of HindIII restriction sites in the pol region as a result of several nucleotide substitutions. Thus, BL6 melanoma cells produce melanoma-specific ecotropic murine leukemia viruses that encode serologically detectable cell surface MAA. The transfection of BL6 melanoma cells with H-2K(b) or H-2K(d) genes but not H-2D(d) or H-2L(d) genes resulted in a loss of MAA expression that was attributed to the changes in proviral DNA and loss of melanoma-specific ecotropic retrovirus particle production. C1 UNIV PITTSBURGH,INST CANC,PITTSBURGH,PA 15213. UNIV PITTSBURGH,DEPT PATHOL,PITTSBURGH,PA 15213. US FDA,CTR BIOL EVALUAT & RES,DIV VIRAL PROD,BETHESDA,MD 20892. NCI,NIH,CELL BIOL LAB,BETHESDA,MD 20892. FU NCI NIH HHS [CA59903] NR 37 TC 16 Z9 16 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 0008-5472 J9 CANCER RES JI Cancer Res. PD OCT 1 PY 1996 VL 56 IS 19 BP 4464 EP 4474 PG 11 WC Oncology SC Oncology GA VK307 UT WOS:A1996VK30700034 PM 8813142 ER PT J AU Evarts, RP Hu, ZY Omori, N Omori, M Marsden, ER Thorgeirsson, SS AF Evarts, RP Hu, ZY Omori, N Omori, M Marsden, ER Thorgeirsson, SS TI Precursor-product relationship between oval cells and hepatocytes: Comparison between tritiated thymidine and bromodeoxyuridine as tracers SO CARCINOGENESIS LA English DT Article ID LIVER STEM-CELLS; RAT-LIVER; DIFFERENTIATION; 5-BROMO-2'-DEOXYURIDINE; 2-ACETYLAMINOFLUORENE; HEPATOCARCINOGENESIS; PROLIFERATION; ACTIVATION; EXPRESSION; GROWTH AB The expansion and differentiation of oval cells in the acetylaminofluorene (AAF)/partial hepatectomy (PH) model was studied utilizing pulse-chase labeling with both tritiated thymidine ([H-3]TdR) and bromodeoxyuridine (BUdR), Animals in which a significant decrease in serum albumin and increase in alanine aminotransferase and bilirubin were observed demonstrated the most prominent differentiation of oval cells into hepatocytes. Administration of [H-3]TdR or BUdR, either individually or together, to the animals on day 6 after partial hepatectomy resulted in labeling of the majority of the oval cells by days 7 and 9 after PH, A striking difference in the distribution of [H-3]TdR- and BUdR-labeled cells in the double labeling experiments was observed on day 11, at which time the number of [H-3]TdR-labeled cells increased 6-fold and that of double labeled cells decreased 2-fold, Furthermore, on day 11 the basophilic foci were weakly positive for BUdR and negative at later time points in animals receiving BUdR alone or together with [H-3]TdR, In contrast, the cells in basophilic foci as well as transitional cells were positive for [H-3]TdR, Cells heavily labeled with both [H-3]TdR and BUdR were present at all time points, indicating an inhibition of the proliferative activity, Pulse labeling of rat liver epithelial cells with BUdR in vitro demonstrated that immunodetection of BUdR was lost after three or more cell divisions. We conclude that the BUdR tagging method is particularly sensitive to label dilution during cell cycling and may not be suitable for establishment of a precursor-product relationship between cell lineages when the progenitor population proliferates more than three times. RP Evarts, RP (reprint author), NCI,DIV BASIC SCI,EXPT CARCINOGENESIS LAB,BLDG 37,ROOM 3C28,37 CONVENT DR MSC4255,BETHESDA,MD 20892, USA. NR 43 TC 54 Z9 58 U1 0 U2 0 PU OXFORD UNIV PRESS UNITED KINGDOM PI OXFORD PA WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP SN 0143-3334 J9 CARCINOGENESIS JI Carcinogenesis PD OCT PY 1996 VL 17 IS 10 BP 2143 EP 2151 DI 10.1093/carcin/17.10.2143 PG 9 WC Oncology SC Oncology GA VM805 UT WOS:A1996VM80500007 PM 8895481 ER PT J AU Sorensen, IK Mortensen, A Kristiansen, E vanKreijl, C Adamson, RH Thorgeirsson, SS AF Sorensen, IK Mortensen, A Kristiansen, E vanKreijl, C Adamson, RH Thorgeirsson, SS TI Short-term carcinogenicity testing of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and 2-amino-3-methylimidazo[4,5-f] quinoline (IQ) in E mu-pim-1 transgenic mice SO CARCINOGENESIS LA English DT Article ID HETEROCYCLIC AMINES; BROILED SARDINE; BEEF EXTRACT; COOKED FOOD; DNA; IDENTIFICATION; ACTIVATION; LYMPHOMAGENESIS; MUTAGENICITY; INDUCTION AB The usefulness of transgenic E mu-pim-1 mice over-expressing the pim-1 oncogene in lymphoid tissues, as sensitive test organisms was studied in a short-term carcinogenicity study. The mice were fed standard diet Altromin 1314 supplemented either with 0.03% 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) for 7 months or with 0.03% 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) for 6 months, PhIP and IQ are heterocyclic amines formed during cooking of meat and fish and are mutagenic to bacteria and cultured mammalian cells, PhIP is a potent mouse lymphomagen, while IQ is a liver carcinogen and also causes lung tumors and tumors of the forestomach in mice. We found that transgenic E mu-pim-1 mice are highly susceptible to PhIP induced lymphomagenesis but do not respond to the IQ treatment, PhIP feeding of E mu-pim-1 mice not only increased the total number of T-cell lymphomas but also decreased the latency time compared to either transgenic or wild-type controls, The effect was most pronounced in the treated female E mu-pim-1 mice, which showed a higher incidence of PhIP induced T-cell lymphomas than transgenic males and a strongly reduced latency period after PhIP treatment compared to non-transgenic mice. Our results suggest that the transgenic E mu-pim-1 mouse may be a useful model for short-term carcinogenicity screening of potential genotoxic carcinogens having the lymphoid system as target tissue, The carcinogen IQ which does not have the lymphoid system as a target was not recognized in this model. C1 NATL INST PUBL HLTH & ENVIRONM,NL-3720 BA BILTHOVEN,NETHERLANDS. NCI,NIH,BETHESDA,MD 20892. RP Sorensen, IK (reprint author), NATL FOOD AGCY,INST TOXICOL,MORKHOJ BYGADE 19,DK-2860 SOBORG,DENMARK. NR 27 TC 16 Z9 16 U1 0 U2 0 PU OXFORD UNIV PRESS UNITED KINGDOM PI OXFORD PA WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP SN 0143-3334 J9 CARCINOGENESIS JI Carcinogenesis PD OCT PY 1996 VL 17 IS 10 BP 2221 EP 2227 DI 10.1093/carcin/17.10.2221 PG 7 WC Oncology SC Oncology GA VM805 UT WOS:A1996VM80500018 PM 8895492 ER PT J AU Einolf, HJ Amin, S Yagi, H Jerina, DM Baird, WM AF Einolf, HJ Amin, S Yagi, H Jerina, DM Baird, WM TI Benzo[c]phenanthrene is activated to DNA-binding diol epoxides in the human mammary carcinoma cell line MCF-7 but only limited activation occurs in mouse skin SO CARCINOGENESIS LA English DT Article ID TUMOR-INITIATING ACTIVITY; POLYCYCLIC AROMATIC-HYDROCARBONS; FEMALE CD RATS; FJORD-REGION; MAMMALIAN-CELLS; STEREOSELECTIVE METABOLISM; ABSOLUTE-CONFIGURATION; MUTAGENIC SPECIFICITY; POTENT CARCINOGEN; LIVER MICROSOMES AB Benzo[c]phenanthrene (B[c]Ph) is an environmental contaminant with low carcinogenic activity in rodent bioassays, B[c]Ph-3,4-diol-1,2-epoxides (B[c]PhDE), however, are among the most tumorigenic diol epoxides known, To determine whether human cells are capable of activating B[c]Ph to DNA-binding metabolites, cultures of the human mammary cell line, MCF-7, were exposed to 10 mu M B[c]Ph for 48, 72 and 96 h or to 1 mu M [(+/-)-B[c]Ph-3,4-dihydrodiol for 48 h. The B[c]Ph-DNA adducts were analyzed by P-33-postlabeling and reverse-phase HPLC. The major B[c]Ph-DNA adducts were formed by the trans-addition of(4R,3S)-dihydroxy-(2S,1R)-epoxy-1,2,3,4-tetrahydro-B[c]Ph to deoxyadenosine [(-)-B[c]PhDE-2dA(t)] and by the cis- and trans-addition of (4S,3R)-dihydroxy-(2S,1R)-epoxy-1,2,3,4-tetrahydro-B[c]Ph to deoxyadenosine [(-)-B[c]PhDE-2dA(c)] and (+)-B[c]PhDE-1d(t)]. Smaller amounts of the trans-addition of (-)-B[c]PhDE-2 were bound to deoxyguanosine. To determine whether B[c]Ph can be metabolically activated to diol epoxides in mouse epidermis, female SENCAR mice were treated topically with 2 mu mol B[c]Ph for 24, 48 or 72 h or with 0.4 mu mol (+/-)-B[c]Ph-3,4-dihydrodiol for 24 or 48 h, In B[c]Ph-treated mice, only small amounts of three B[c]PhDE-DNA adducts were detected B[c]PhDE-2dA(t), (+)-B[c]PhDE-1dA(t) and (+)-B[c]PhDE-1dA(c)] at 24, 48 and 72 h. In contrast, mice treated topically with 0.4 mu mol(+/-)-B[c]Ph-3,4-dihydrodiol formed B[c]PhDE-DNA adducts at levels 6-fold greater than those observed with B[c]Ph at 48 h. The higher formation of B[c]PhDE-DNA adducts by (+/-)-B[c]Ph-3,4-dihydrodiol correlates with the greater potency of (+/-)-B[c]Ph-3,4-dihydrodiol than of B[c]Ph as a tumor initiator in mouse skin, The low extent of formation of B[c]PhDE from B[c]Ph in mouse epidermis may explain the low tumorigenicity of B[c]Ph in this tissue, These results indicate activation of B[c]Ph in mouse skin and tumorigenesis results in that tissue may not adequately assess the potential capability cells from humans to activate B[c]Ph to ultimate carcinogenic metabolites. C1 PURDUE UNIV,DEPT MED CHEM & MOL PHARMACOL,W LAFAYETTE,IN 47907. AMER HLTH FDN,DIV CHEM CARCINOGENESIS,VALHALLA,NY 10595. NIDDKD,BIOORGAN CHEM LAB,NIH,BETHESDA,MD 20892. FU NCI NIH HHS [CA28825, CA40228] NR 48 TC 32 Z9 32 U1 0 U2 0 PU OXFORD UNIV PRESS UNITED KINGDOM PI OXFORD PA WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP SN 0143-3334 J9 CARCINOGENESIS JI Carcinogenesis PD OCT PY 1996 VL 17 IS 10 BP 2237 EP 2244 DI 10.1093/carcin/17.10.2237 PG 8 WC Oncology SC Oncology GA VM805 UT WOS:A1996VM80500020 PM 8895494 ER PT J AU Fleg, JL AF Fleg, JL TI Doppler echocardiography and prognosis in the elderly SO CARDIOLOGY IN THE ELDERLY LA English DT Editorial Material DE Doppler echocardiography; prognosis; elderly ID LEFT-VENTRICULAR MASS; VALVULAR AORTIC-STENOSIS; VALVE REPLACEMENT; BLOOD-PRESSURE; HEART-DISEASE; POPULATION; MEN; AGE; PREVALENCE; HYPERTROPHY RP Fleg, JL (reprint author), NIA,GERONTOL RES CTR,CARDIOVASC SCI LAB,4940 EASTERN AVE,BALTIMORE,MD 21224, USA. NR 28 TC 0 Z9 0 U1 0 U2 0 PU RAPID SCIENCE PUBLISHERS PI LONDON PA 2-6 BOUNDARY ROW, LONDON, ENGLAND SE1 8NH SN 1058-3661 J9 CARDIOL ELDER JI Cardiol. Elder. PD OCT-DEC PY 1996 VL 4 IS 5-6 BP 219 EP 222 PG 4 WC Cardiac & Cardiovascular Systems; Geriatrics & Gerontology SC Cardiovascular System & Cardiology; Geriatrics & Gerontology GA WT720 UT WOS:A1996WT72000007 ER PT J AU Lane, NJ Balbo, A Rapoport, SI AF Lane, NJ Balbo, A Rapoport, SI TI A fine structural study of the hippocampus and dorsal root ganglion in mouse trisomy 16, a model of Down's syndrome SO CELL BIOLOGY INTERNATIONAL LA English DT Article DE hippocampus; model for Down's syndrome; Alzheimer's disease; mouse trisomy 16; neuropathology; dorsal root ganglion ID ALZHEIMERS-DISEASE NEUROPATHOLOGY; HUMAN CHROMOSOME-21; NEURONS; TRANSPLANTS; FETUS; MICE; ABNORMALITIES; PATHOLOGY; DEMENTIA; SYSTEMS AB Mouse trisomy 16 (Ts16) appears to provide an animal model of Down's syndrome in that a portion of mouse chromosome 16 is syntenic with part of human chromosome 21. Trisomy 21 in human beings leads to the mental retardation of Down's syndrome and in middle age, to some presenile anatomic and clinical features of Alzheimer's disease. Neural tissue from aging Ts16 mice is unavailable, however, as Ts16 mouse embryos die late in utero. We studied these embryos looking at the ultrastructure of neurons from the hippocampus and dorsal root ganglion in normal control mice embryos (diploid) and in Ts16 late embryonic litter mates after day 15 of gestation. The organelles in the Ts16 neurons looked similar to those in control neurons, fixed and processed under similar conditions. No obvious neuropathological structures were observed. These results, when compared to reports on electrophysiological abnormalities of cultured fetal Ts16 neurons and on abnormalities in neurotransmitter markers in the Ts16 fetal brain, lead us to suggest that the mental retardation of Down's syndrome is likely to result from functional and chemical defects not directly related to abnormal neuronal ultrastructure. When related to fine structural studies of transplanted embryonic Ts16 hippocampus which have been maintained for long periods of time, these results indicate that the trisomic mouse brain would not be useful as a structural model for Down's syndrome and hence presenile Alzheimer's disease, as it is not associated with any detectable morphological abnormality. (C) 1996 Academic Press Limited C1 NIA, NEUROSCI LAB, NIH, BETHESDA, MD 20892 USA. DEPT ZOOL, CAMBRIDGE CB2 3EJ, ENGLAND. FU Wellcome Trust NR 38 TC 11 Z9 11 U1 0 U2 0 PU WILEY-BLACKWELL PI HOBOKEN PA 111 RIVER ST, HOBOKEN 07030-5774, NJ USA SN 1065-6995 EI 1095-8355 J9 CELL BIOL INT JI Cell Biol. Int. PD OCT PY 1996 VL 20 IS 10 BP 673 EP 680 DI 10.1006/cbir.1996.0089 PG 8 WC Cell Biology SC Cell Biology GA VV585 UT WOS:A1996VV58500003 PM 8969460 ER PT J AU Malliri, A Yeudall, A Nikolic, M Crouch, DH Parkinson, EK Ozanne, B AF Malliri, A Yeudall, A Nikolic, M Crouch, DH Parkinson, EK Ozanne, B TI Sensitivity to transforming growth factor beta 1-induced growth arrest is common in human squamous cell carcinoma cell lines: c-MYC down-regulation and p21(waf1) induction are important early events SO CELL GROWTH & DIFFERENTIATION LA English DT Article ID TGF-BETA RECEPTOR; CYCLE ARREST; II RECEPTOR; MURINE KERATINOCYTES; GENE-TRANSCRIPTION; TUMOR SUPPRESSION; EPITHELIAL-CELLS; BINDING DOMAINS; CDK INHIBITORS; CANCER CELLS AB Transforming growth factor beta 1 (TGF-beta 1) is a potent inhibitor of keratinocyte proliferation and a potential tumor suppressor of squamous cell carcinomas (SCCs), TGF-beta 1 exerts its antiproliferative effects by inhibiting key transitions required for progression from G(1) to the S phase of the cell cycle, exemplified by a rapid reduction of c-MYC and inhibition of the G(1) cyclin/cyclin-dependent kinases by induction of their inhibitors p21(waf1), p27(Kip1), and p15(INK4B). A significant majority of a new series of human SCC cell lines were found to be as sensitive as primary human epidermal keratinocytes to TGF-beta 1 growth inhibition, Only a minority of cell lines derived from late-stage tumors were resistant. An early and rapid increase in p21(Waf1) and reduction in c-MYC protein levels were important concomitants for TGF-beta 1 growth inhibition; these changes occurred exclusively in each of the sensitive cell lines. Expression of p15(INK4B) was found to be neither necessary nor sufficient for TGF-beta 1 growth arrest in the sensitive and resistant cell lines, respectively. TGF-beta 1 induced alterations in other cell cycle regulatory molecules, cyclin-dependent kinase 4, cyclin D1, pRB, and p27(Kip1), occurred late and were dispensable in some of the sensitive cell lines. Expression of exogenous mycER fusion protein in one of the sensitive cell lines did not render the cells resistant to TGF-beta 1-induced growth arrest nor prevent p21(waf1) induction or down-regulation of both c-MYC and mycER proteins, However, in TGF-beta 1-resistant subclones of sensitive mycER-expressing cells, p21(waf1) was not induced, whereas both c-MYC and mycER protein levels decreased following TGF-beta 1 treatment. We conclude that TGF-beta 1 activates multiple cell cycle inhibitory pathways dependent upon p21(waf1) induction and c-MYC degradation and that it does not function as a tumor suppressor in the majority of SCCs. C1 BEATSON INST CANC RES,CRC BEATSON LABS,GLASGOW G61 1BD,LANARK,SCOTLAND. NIDR,CELLULAR DEV & ONCOL LAB,MOL CARCINOGENESIS GRP,BETHESDA,MD 20892. UNIV DUNDEE,NINEWELLS HOSP & MED SCH,CTR BIOMED RES,DUNDEE DD1 9SY,SCOTLAND. NR 67 TC 58 Z9 59 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 1044-9523 J9 CELL GROWTH DIFFER JI Cell Growth Differ. PD OCT PY 1996 VL 7 IS 10 BP 1291 EP 1304 PG 14 WC Cell Biology SC Cell Biology GA VL649 UT WOS:A1996VL64900002 PM 8891333 ER PT J AU Kwon, TK Buchholz, MA Chrest, FJ Nordin, AA AF Kwon, TK Buchholz, MA Chrest, FJ Nordin, AA TI Staurosporine-induced G(1) arrest is associated with the induction and accumulation of cyclin-dependent kinase inhibitors SO CELL GROWTH & DIFFERENTIATION LA English DT Article ID CELL-CYCLE; RETINOBLASTOMA PROTEIN; G1 PHASE; TRANSFORMED-CELLS; POTENT INHIBITOR; MAMMALIAN-CELLS; CDK INHIBITORS; PHOSPHORYLATION; PROGRESSION; LYMPHOCYTES AB The addition of 10 nM staurosporine (ST) to MDA 361 breast carcinoma cells induces a G(1) arrest, which correlates with the loss of the catalytic activity of the G(1)-associated cyclin-dependent kinases (cdks) and increased levels of underphosphorylated retinoblastoma protein. This treatment resulted in a slight but detectable reduction in the protein levels of cdk6 but did not reduce the levels of cdk2, cdk4, or the D cyclins. The level of cyclin E declined initially but returned to normal levels 24 h after exposure to 10 nM ST. Because the levels of the G(1) cdks and cyclins did not correlate with loss of kinase activity, the role of the cdk inhibitors involved in regulating the activity of the G(1)-associated cdks was investigated. The significant reduction in cdk activity observed in MDA 361 cells treated with ST for 24 h correlated with increased levels of p18 and p27(Kip). The inhibition of kinase activity of preformed cdk2 complexes by lysates of MDA 361 cells that had been treated with 10 nM ST for 24 h was shown to be due to p27(Kip). The reduction in the level of the active phosphorylated form of cdk2 also correlated with an increase in the level of p27(Kip), which has been shown to inhibit the phosphorylation of the activating Thr-160 residue of cdk2. These results indicate that treatment of MDA 361 cells with 10 nM ST induces a significant increase in the levels of several cdk inhibitors that appear to be responsible for the observed G(1) arrest. C1 NIA,NIH,CTR GERONTOL RES,LAB CLIN PHYSIOL,BALTIMORE,MD 21224. NR 45 TC 32 Z9 32 U1 0 U2 1 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 1044-9523 J9 CELL GROWTH DIFFER JI Cell Growth Differ. PD OCT PY 1996 VL 7 IS 10 BP 1305 EP 1313 PG 9 WC Cell Biology SC Cell Biology GA VL649 UT WOS:A1996VL64900003 PM 8891334 ER PT J AU Bourdi, M Chen, WQ Peter, RM Martin, JL Buters, JTM Nelson, SD Pohl, LR AF Bourdi, M Chen, WQ Peter, RM Martin, JL Buters, JTM Nelson, SD Pohl, LR TI Human cytochrome P450 2E1 is a major autoantigen associated with halothane hepatitis SO CHEMICAL RESEARCH IN TOXICOLOGY LA English DT Article ID ENDOPLASMIC-RETICULUM AUTOANTIBODIES; PROTEIN DISULFIDE-ISOMERASE; PRIMARY BILIARY-CIRRHOSIS; SERUM ANTIBODIES; LIVER CYTOCHROME-P-450; CONFORMATIONAL EPITOPE; ADDISONS-DISEASE; IGG SUBCLASS; B-CELLS; INDUCTION AB Autoantibodies against specific human cytochrome P450s have been found in the sera of patients suffering from a variety of diseases, including those caused by drugs. in the cases of tienilic acid- and dihydralazine-induced hepatitis, patients have serum autoantibodies directed against cytochromes P450 2C9 and P450 1A2, respectively. In the present study, we have found that 25 of 56 (45%) patients diagnosed with halothane hepatitis have autoantibodies that react with human cytochrome P450 2E1 that was purified from a baculovirus expression system. The autoantibodies inhibited the activity of cytochrome P450 2E1 and appeared to be directed against mainly conformational epitopes. In addition, because cytochrome P450 2E1 became trifluoroacetylated when it oxidatively metabolized halothane, it is possible that the covalently altered form of cytochrome P450 2E1 may be able to bypass the immunologic tolerance that normally exists against cytochrome P450 2E1. A similar mechanism may explain the formation of autoantibodies that have been found against other cellular targets of the reactive trifluoroacetyl chloride metabolite of halothane. C1 UNIV WASHINGTON,DEPT MED CHEM,SEATTLE,WA 98195. JOHNS HOPKINS MED INST,DEPT ANESTHESIOL & CRIT CARE MED,BALTIMORE,MD 21287. NCI,MOL CARCINOGENESIS LAB,NIH,BETHESDA,MD 20892. RP Bourdi, M (reprint author), NHLBI,LAB MOL IMMUNOL,MOL & CELLULAR TOXICOL SECT,BLDG 10,ROOM 8N110,BETHESDA,MD 20892, USA. RI Buters, Jeroen/G-5070-2011 FU NIEHS NIH HHS [ES 02728]; NIGMS NIH HHS [GM 32165] NR 57 TC 124 Z9 128 U1 0 U2 1 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 SN 0893-228X J9 CHEM RES TOXICOL JI Chem. Res. Toxicol. PD OCT-NOV PY 1996 VL 9 IS 7 BP 1159 EP 1166 DI 10.1021/tx960083q PG 8 WC Chemistry, Medicinal; Chemistry, Multidisciplinary; Toxicology SC Pharmacology & Pharmacy; Chemistry; Toxicology GA VN272 UT WOS:A1996VN27200014 PM 8902272 ER PT J AU Thadani, PV AF Thadani, PV TI NIDA conference report on cardiopulmonary complications of ''crack'' cocaine use - Clinical manifestations and pathophysiology SO CHEST LA English DT Article DE cocaine; crack; cardiovascular; pulmonary; complications; pathophysiology; abused drugs; biological mechanisms ID PLASMA CHOLINESTERASE ACTIVITY; PULMONARY COMPLICATIONS; INTRAVENOUS COCAINE; VASOCONSTRICTION; TOXICITY; ABUSERS; ASTHMA; DRUG; RATS RP Thadani, PV (reprint author), NIDA,DIV BASIC RES,1-A-19,5600 FISHERS LANE,ROCKVILLE,MD 20857, USA. NR 42 TC 21 Z9 22 U1 0 U2 0 PU AMER COLL CHEST PHYSICIANS PI NORTHBROOK PA 3300 DUNDEE ROAD, NORTHBROOK, IL 60062-2348 SN 0012-3692 J9 CHEST JI Chest PD OCT PY 1996 VL 110 IS 4 BP 1072 EP 1076 DI 10.1378/chest.110.4.1072 PG 5 WC Critical Care Medicine; Respiratory System SC General & Internal Medicine; Respiratory System GA VN269 UT WOS:A1996VN26900039 PM 8874270 ER PT J AU Harwood, RL Schoelmerich, A VenturaCook, E Schulze, PA Wilson, SP AF Harwood, RL Schoelmerich, A VenturaCook, E Schulze, PA Wilson, SP TI Culture and class influences on angle and Puerto Rican mothers' beliefs regarding long-term socialization goals and child behavior SO CHILD DEVELOPMENT LA English DT Article ID MODELS; STYLE AB These 2 studies examine culture and socioeconomic status as simultaneous possible sources for group differences in mothers' beliefs regarding desirable and undesirable long-term socialization goals and child behavior. In Study 1, 100 mothers of young toddlers aged 12-24 months from 5 sociocultural groups participated: middle- and lower-class Angle, middle- and lower-class island Puerto Rican, and lower-class migrant Puerto Rican. Results indicate that culture and socioeconomic status contribute independently to group differences, but that cultural effects appear to be stronger. Study 2 examined cultural differences in perceptions of behaviors using middle-class Angle and Puerto Rican mothers only. The findings support those of Study 1, suggesting that Angle and Puerto Rican mothers place differential value on the constructs of Self-Maximization and Proper Demeanor, even when socioeconomic status is controlled for. The findings of these studies have important implications for the culturally sensitive study of the relation between parental beliefs and behaviors. C1 NIH,BETHESDA,MD 20892. UNIV NEW ORLEANS,NEW ORLEANS,LA 70148. RP Harwood, RL (reprint author), UNIV CONNECTICUT,SCH FAMILY STUDIES,U-58,348 MANSFIELD RD,STORRS,CT 06269, USA. RI Schoelmerich, Axel/C-9039-2009 OI Schoelmerich, Axel/0000-0002-9844-3920 FU NIMH NIH HHS [MH 09911] NR 24 TC 59 Z9 68 U1 1 U2 2 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 0009-3920 J9 CHILD DEV JI Child Dev. PD OCT PY 1996 VL 67 IS 5 BP 2446 EP 2461 DI 10.1111/j.1467-8624.1996.tb01867.x PG 16 WC Psychology, Educational; Psychology, Developmental SC Psychology GA WE655 UT WOS:A1996WE65500034 PM 9022250 ER PT J AU ZahnWaxler, C Friedman, RJ Cole, PM Mizuta, I Hiruma, N AF ZahnWaxler, C Friedman, RJ Cole, PM Mizuta, I Hiruma, N TI Japanese and United States preschool children's responses to conflict and distress SO CHILD DEVELOPMENT LA English DT Article ID BEHAVIOR; THAI AB Japanese and U.S. preschool children's responses to hypothetical interpersonal dilemmas were examined as a function of culture, gender, and maternal child-rearing values. U.S. children showed more anger, more aggressive behavior and language, and underregulation of emotion than Japanese children, across different contexts of assessment. Children from the 2 cultures appeared more similar on prosocial and avoidant patterns, though in some contexts U.S. children also showed more prosocial themes. Girls from both cultures expressed more prosocial themes and sometimes more anger than boys. Maternal encouragement of children's emotional expressivity was correlated with anger and aggression in children. It was more characteristic of U.S. than Japanese mothers, while emphasis on psychological discipline (reasoning; guilt and anxiety induction) was more characteristic of Japanese than U.S. mothers. The relevance of a conceptual framework that focuses on differences in Eastern and Western cultures in self-construals regarding independence and interdependence is considered. C1 ARIZONA STATE UNIV,TEMPE,AZ 85287. PENN STATE UNIV,UNIVERSITY PK,PA 16802. OSAKA UNIV,SCH MED,SUITA,OSAKA 565,JAPAN. KEIO UNIV,SCH MED,TOKYO 108,JAPAN. RP ZahnWaxler, C (reprint author), NIMH,BETHESDA,MD 20892, USA. NR 30 TC 76 Z9 78 U1 1 U2 11 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 0009-3920 J9 CHILD DEV JI Child Dev. PD OCT PY 1996 VL 67 IS 5 BP 2462 EP 2477 DI 10.1111/j.1467-8624.1996.tb01868.x PG 16 WC Psychology, Educational; Psychology, Developmental SC Psychology GA WE655 UT WOS:A1996WE65500035 PM 9022251 ER PT J AU Pedersen, FA Huffman, LC delCarmen, R Bryan, YE AF Pedersen, FA Huffman, LC delCarmen, R Bryan, YE TI Prenatal maternal reactivity to infant cries predicts postnatal perceptions of infant temperament and marriage appraisal SO CHILD DEVELOPMENT LA English DT Article ID ABUSE POTENTIAL INVENTORY; ACOUSTIC FEATURES; MARITAL CHANGE; RESPONSES; TRANSITION; PARENTHOOD; DIFFICULTNESS; ADJUSTMENT; DEPRESSION; PREGNANCY AB In a sample of 60 primiparous women, cardiac response and ratings of subjective aversiveness to recordings of unfamiliar infant cries were studied at 32 weeks' gestation. Regression analyses were used to examine relations between cardiac acceleration and subjective aversiveness and 3 groups of postnatal dependent variables: perception of infant temperament, the mother's emotional state, and her appraisal of her marriage. Mothers who prenatally rated the cry recordings as more aversive postnatally described their 3-month-old infants as more fussy/difficult and unpredictable. With statistical control for prenatal variation on the emotional state and marital outcome measures, cardiac acceleration predicted later marital quality. Women who showed greater cardiac acceleration to the cries described their postnatal marital relationships more negatively. C1 NICHHD,BETHESDA,MD. NR 46 TC 8 Z9 8 U1 1 U2 5 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 0009-3920 J9 CHILD DEV JI Child Dev. PD OCT PY 1996 VL 67 IS 5 BP 2541 EP 2552 PG 12 WC Psychology, Educational; Psychology, Developmental SC Psychology GA WE655 UT WOS:A1996WE65500040 PM 9022255 ER PT J AU Lowe, X OHogan, S Moore, D Bishop, J Wyrobek, A AF Lowe, X OHogan, S Moore, D Bishop, J Wyrobek, A TI Aneuploid epididymal sperm detected in chromosomally normal and Robertsonian translocation-bearing mice using a new three-chromosome FISH method SO CHROMOSOMA LA English DT Article ID IN-SITU HYBRIDIZATION; EMBRYOS FERTILIZED INVITRO; METACENTRIC CHROMOSOMES; MOUSE; CELLS AB We present a new method to detect epididymal sperm aneuploidy (ESA) in mice using simultaneous fluorescence in situ hybridization (FISH) with DNA probes specific for mouse chromosomes X, Y and 8. The method was applied to Robertsonian (Rb) translocation (8.14) heterozygotes and homozygotes as well as the chromosomally normal B6C3F1. The sex ratios of sperm did not differ from the expected 1:1 and the hybridization efficiencies were similar to 99.7% for over 60 000 sperm analyzed. Mice heterozygous for Rb (8.14) produced about tenfold higher rates of sperm with chromosome 8 hyperhaploidy than did Rb (8.14) homozygotes or chromosomally normal mice, while frequencies of sperm with hyperhaploidies for chromosomes X and Y were unaffected in all three lines of mice. Hyperhaploid frequencies obtained with the ESA method were consistent with those of the previous testicular FISH method and were validated by published data obtained by conventional cytogenetic analyses (meiotic metaphase II and first cleavage). Thus, the mouse three-chromosome ESA assay together with the previously developed aneuploidy assay for human sperm constitute a promising pair of interspecific biomarkers for comparative studies of the genetic and physiologic mechanisms of the induction and persistence of aneuploidy in male germ cells. C1 LAWRENCE LIVERMORE NATL LAB,BIOL & BIOTECHNOL RES PROGRAM,LIVERMORE,CA 94550. NIEHS,RES TRIANGLE PK,NC 27709. FU NIEHS NIH HHS [IAG Y01-ES-10203-00] NR 39 TC 30 Z9 31 U1 0 U2 0 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 SN 0009-5915 J9 CHROMOSOMA JI Chromosoma PD OCT PY 1996 VL 105 IS 4 BP 204 EP 210 PG 7 WC Biochemistry & Molecular Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Genetics & Heredity GA VT553 UT WOS:A1996VT55300002 PM 8854879 ER PT J AU Stone, PH Chaitman, BR McMahon, RP Andrews, TC MacCallum, G Sharaf, B Frishman, W Deanfield, JE Sopko, G Pratt, C Goldberg, AD Rogers, WJ Hill, J Proschan, M Pepine, CJ Bourassa, HG Conti, CR AF Stone, PH Chaitman, BR McMahon, RP Andrews, TC MacCallum, G Sharaf, B Frishman, W Deanfield, JE Sopko, G Pratt, C Goldberg, AD Rogers, WJ Hill, J Proschan, M Pepine, CJ Bourassa, HG Conti, CR TI Asymptomatic Cardiac Ischemia Pilot (ACIP) study - Relationship between exercise-induced and ambulatory ischemia in patients with stable coronary disease SO CIRCULATION LA English DT Article DE ischemia; electrocardiography; coronary disease; prognosis; trials ID SILENT-MYOCARDIAL-ISCHEMIA; ANGINA-PECTORIS; DAILY-LIFE; ARTERY DISEASE; SEVERITY; FREQUENCY; THERAPY AB Background We investigated whether the presence and frequency of asymptomatic ischemic episodes recorded during ambulatory ECG (AECG) monitoring could be predicted on the basis of clinical characteristics or exercise treadmill test (ETT) performance in patients with stable coronary disease and whether the estimate of ischemia severity was similar between the AECG and ETT. Methods and Results Patients screened for the Asymptomatic Cardiac Ischemia Pilot (ACIP) study were selected for the current analysis if data were available from 48-hour AECG monitoring as well as from an ETT during which the patient developed greater than or equal to 1-mm ST-segment depression. Exercise ECG data were available for 143 of the 910 patients without ischemic episodes and for 659 of the 910 patients with ischemic episodes during AECG monitoring. Angina was more frequent among patients with ambulatory ischemic episodes than among patients without such ischemia (P<.001). Patients with AECG ischemia had a consistently more marked ischemic response on the ETT than patients without AECG ischemia; patients likely to have AECG ischemia could be predicted on the basis of ETT performance characteristics. However, the correlation coefficients between the severity of ischemia estimated by ETT and by AECG were small. Conclusions There are significant relations between ischemia detected by AECG monitoring and by ETT, but the relations are limited, indicating that the two tests are not redundant to characterize coronary patients. A larger study investigating the prognostic significance of the ischemia identified by each modality, with follow-up for clinical events, will be necessary to determine the most appropriate methods to evaluate patients with stable coronary disease. C1 ST LOUIS UNIV, MED CTR, DEPT INTERNAL MED, DIV CARDIOL, ST LOUIS, MO USA. MARYLAND MED RES INST, ACIP CORRDINATING CTR, BALTIMORE, MD USA. WILFORD HALL USAF MED CTR, PSMC, DEPT CARDIOL, AETC, LACKLAND AFB, TX USA. UNIV RHODE ISL, PROVIDENCE, RI 02908 USA. ALBERT EINSTEIN HOSP, BRONX, NY USA. ST BARTHOLOMEWS HOSP, DEPT CARDIOL, LONDON, ENGLAND. NIH, BETHESDA, MD 20892 USA. BAYLOR COLL MED, CARDIOL SECT, HOUSTON, TX 77030 USA. HENRY FORD HOSP, DETROIT, MI 48202 USA. UNIV ALABAMA, MED CTR, BIRMINGHAM, AL 35294 USA. UNIV FLORIDA, GAINESVILLE, FL USA. VET ADM MED CTR, CARDIOL SECT, GAINESVILLE, FL 32602 USA. INST CARDIOL, MONTREAL, PQ, CANADA. RP Stone, PH (reprint author), BRIGHAM & WOMENS HOSP, DIV CARDIOVASC, 75 FRANCIS ST, BOSTON, MA 02115 USA. RI Deanfield, John/C-5178-2008; McMahon, Robert/C-5462-2009; OI Bourassa, Martial G./0000-0002-4439-8650 FU NHLBI NIH HHS [HV-90-08, HV-90-07, HV-91-05] NR 34 TC 18 Z9 19 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 OCT 1 PY 1996 VL 94 IS 7 BP 1537 EP 1544 PG 8 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA VJ979 UT WOS:A1996VJ97900010 PM 8840841 ER PT J AU Baluna, R Sausville, EA Stone, MJ StetlerStevenson, MA Uhr, JW Vitetta, ES AF Baluna, R Sausville, EA Stone, MJ StetlerStevenson, MA Uhr, JW Vitetta, ES TI Decreases in levels of serum fibronectin predict the severity of vascular leak syndrome in patients treated with ricin A chain-containing immunotoxins SO CLINICAL CANCER RESEARCH LA English DT Article ID PHASE-I EVALUATION; B-CELL LYMPHOMA; PLASMA FIBRONECTIN; CONTINUOUS-INFUSION; ENDOTHELIAL-CELLS; CLINICAL-TRIALS; A-CHAIN; GM-CSF; INTERLEUKIN-2; THERAPY AB The major dose-limiting adverse effect of ricin A chain-containing immunotoxin (IT) therapy is vascular leak syndrome (VLS), Since plasma fibronectin (Fn) plays a role in maintaining microcirculatory integrity and since the gradient between plasma and tissue Fn can be altered in various pathological situations, we determined whether the administration of IT-ricin A chain to patients resulted in changes in the levels of serum Fn and, if so, whether these changes correlated with the severity of VLS, We also measured the serum levels of tumor necrosis factor alpha. (TNF alpha), a proinflammatory cytokine which has been implicated in tissue damage and in interleukin 2-mediated VLS, Our results indicate that the most severe manifestations of VLS were associated with the highest pretreatment levels of Fn, the largest decreases in Fn immediately after starting IT therapy, increases in the levels of serum TNF alpha, higher concentrations of circulating IT, and the lowest numbers of circulating tumor cells, These parameters should, therefore, be useful for predicting which patients will have severe VLS. C1 UNIV TEXAS,SW MED CTR,CTR CANC IMMUNOBIOL,DALLAS,TX 75235. UNIV TEXAS,SW MED CTR,DEPT MICROBIOL,DALLAS,TX 75235. NCI,DIV CANC TREATMENT DIAG & CTR,DEV THERAPEUT PROGRAM,BETHESDA,MD 20892. NCI,DIV CLIN SCI,PATHOL LAB,BETHESDA,MD 20892. BAYLOR UNIV,MED CTR,DALLAS,TX 75246. CHARLES A SAMMONS CANC CTR,DALLAS,TX 75246. FU NCI NIH HHS [CA-41081, CA-28149] NR 48 TC 12 Z9 12 U1 0 U2 1 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD OCT PY 1996 VL 2 IS 10 BP 1705 EP 1712 PG 8 WC Oncology SC Oncology GA VL234 UT WOS:A1996VL23400010 PM 9816120 ER PT J AU Belotti, D Rieppi, M Nicoletti, MI Casazza, AM Fojo, T Taraboletti, G Giavazzi, R AF Belotti, D Rieppi, M Nicoletti, MI Casazza, AM Fojo, T Taraboletti, G Giavazzi, R TI Paclitaxel (Taxol(R)) inhibits motility of paclitaxel-resistant human ovarian carcinoma cells SO CLINICAL CANCER RESEARCH LA English DT Article ID CANCER-CELLS; MICROTUBULE INHIBITORS; TUMOR-CELLS; MURINE; XENOGRAFTS; MIGRATION; INVASION; THERAPY; AGENT AB The effect of paclitaxel on the adhesive and motility properties of human ovarian carcinoma cell lines was investigated, Paclitaxel significantly inhibited the motility of OVCAR 5, SK-OV-3, and HOC-1OTC ovarian carcinoma cell lines (IC50 = 2.1 x 10(-8), 2 x 10(-9), and 1.9 x 10(-8) M, respectively) but did not affect the adhesion of these cells to the subendothelial matrix, The association between inhibition of motility and cytotoxic activity was investigated using an A2780 subclone (1A9) and three paclitaxel-resistant variants (designated 1A9/PTX22, 1A9/PTX10, and 1A9/PTX18), Although paclitaxel did not significantly affect the adhesion to subendothelial matrix of the sublines, it completely inhibited their migration, Inhibition of migration was similar in 1A9 cells and the resistant sublines, with an IC50 of 1 x 10(-8) for 1A9 cells and 5.4 x 10(-9), 1.1 x 10(-8), and 5.2 x 10(-9) M for 1A9/PTX22, 1A9/PTX10, and 1A9/PTX18, respectively, Paclitaxel inhibited motility induced by soluble attractant (chemotaxis) and immobilized attractant (haptotaxis), Inhibition of cell motility occurred in the absence of an antiproliferative effect, because higher concentrations of paclitaxel were required to inhibit tumor cell proliferation (IC50 = 1.9 x 10(-7) and 4.6 x 10(-6), 1 x 10(-5), and 3.1 x 10(-6) M for 1A9 and 1A9/PTX22, 1A9/PTX10, and 1A9/PTX18, respectively), These data show that paclitaxel is a potent inhibitor of ovarian carcinoma cell motility and that this activity is independent of its cytotoxic activity. C1 MARIO NEGRI INST PHARMACOL RES,I-24125 BERGAMO,ITALY. BRISTOL MYERS SQUIBB PHARMACEUT RES INST,PRINCETON,NJ 08543. NCI,MED BRANCH,NIH,BETHESDA,MD 20892. NR 19 TC 51 Z9 52 U1 0 U2 1 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD OCT PY 1996 VL 2 IS 10 BP 1725 EP 1730 PG 6 WC Oncology SC Oncology GA VL234 UT WOS:A1996VL23400013 PM 9816123 ER PT J AU Trivers, GE DeBenedetti, VMG Cawley, HL Caron, G Harrington, AM Bennett, WP Jett, JR Colby, TV Tazelaar, H Pairolero, P Miller, RD Harris, CC AF Trivers, GE DeBenedetti, VMG Cawley, HL Caron, G Harrington, AM Bennett, WP Jett, JR Colby, TV Tazelaar, H Pairolero, P Miller, RD Harris, CC TI Anti-p53 antibodies in sera from patients with chronic obstructive pulmonary disease can predate a diagnosis of cancer SO CLINICAL CANCER RESEARCH LA English DT Article ID TUMOR-SUPPRESSOR GENE; CELLULAR PROTEIN P53; HUMAN LUNG-CANCER; LARGE T-ANTIGEN; BREAST-CANCER; POOR-PROGNOSIS; BARRETTS-ESOPHAGUS; IMMUNE-RESPONSE; MUTATIONS; EXPRESSION AB Serum anti-p53 antibodies (p53-Abs) may be surrogate markers for both p53 alterations and preclinical cancer. Ancillary to a prospective trial to abate progressive development of clinical stages of chronic obstructive pulmonary disease, we conducted a retrospective, nested case control study, Twenty-three cases were diagnosed with cancer during the trial, Enzyme immunoassay, immunoblotting, and immunoprecipitation were used to detect p53-Abs in serum, immunohistochemistry (IHC) to detect p53 accumulation, and single-strand conformation polymorphism and DNA sequencing to detect p53 mutations in tumor samples, p53-Abs were detected by three types of assays in five (23%) of the cancer patients, 80% of whom had detectable p53-Abs before diagnosis: 2 lung cancers (7 and 6 months before), 1 prostate cancer (11 months), and 1 breast cancer (5 months), Four Ab-positive patients had IHC-positive tumors. Two of 4 Ab-positive patients and 2 of 14 Ab-negative had p53 missense mutations or base pair deletion and LHC-positive tumors, The 44 noncancer COPD controls, matched with the cancer cases for age, gender, and smoking habits, were negative for p53-Abs, These results indicate that p53-Abs may facilitate the early diagnosis of cancer in a subset of smokers with chronic obstructive pulmonary disease who are at an increased cancer risk. C1 MAYO CLIN,ROCHESTER,MN 55905. RP Trivers, GE (reprint author), NCI,HUMAN CARCINOGENESIS LAB,NIH,BLDG 37,ROOM 2C04,BETHESDA,MD 20892, USA. NR 38 TC 92 Z9 93 U1 0 U2 3 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD OCT PY 1996 VL 2 IS 10 BP 1767 EP 1775 PG 9 WC Oncology SC Oncology GA VL234 UT WOS:A1996VL23400018 PM 9816128 ER PT J AU Hruszkewycz, AM Delgado, RM Khan, MA Bennett, WP AF Hruszkewycz, AM Delgado, RM Khan, MA Bennett, WP TI Semiautomated sequence-specific mutation detection of the human K-ras oncogene using ''cold'' SSCP analysis SO CLINICAL CHEMISTRY LA English DT Letter ID POLYMERASE CHAIN-REACTION; CELL LUNG-CANCER; POINT MUTATIONS; GENE-MUTATIONS; PANCREATIC ADENOCARCINOMA; AMPLIFICATION MASA; SHORTENED SURVIVAL; COLORECTAL TUMORS; P53 MUTATIONS; ALLELE C1 NCI,HUMAN CARCINOGENESIS LAB,BETHESDA,MD 20892. RP Hruszkewycz, AM (reprint author), NIH,CLIN CHEM SERV,DEPT CLIN PATHOL,CTR CLIN,BETHESDA,MD 20892, USA. NR 37 TC 2 Z9 2 U1 0 U2 0 PU AMER ASSOC CLINICAL CHEMISTRY PI WASHINGTON PA 2101 L STREET NW, SUITE 202, WASHINGTON, DC 20037-1526 SN 0009-9147 J9 CLIN CHEM JI Clin. Chem. PD OCT PY 1996 VL 42 IS 10 BP 1717 EP 1719 PG 3 WC Medical Laboratory Technology SC Medical Laboratory Technology GA VL761 UT WOS:A1996VL76100026 PM 8855163 ER PT J AU Avgerinos, PC Nieman, LK Oldfield, EH Cutler, GB AF Avgerinos, PC Nieman, LK Oldfield, EH Cutler, GB TI A comparison of the overnight and the standard metyrapone test for the differential diagnosis of adrenocorticotrophin-dependent Cushing's syndrome SO CLINICAL ENDOCRINOLOGY LA English DT Article ID CORTICOTROPIN-RELEASING HORMONE; DEXAMETHASONE SUPPRESSION TEST; STIMULATION TEST; CORTISOL; 11-DEOXYCORTISOL AB OBJECTIVE We wished to develop optimal criteria for interpreting the single-dose overnight metyrapone test and to compare the diagnostic efficiency of the overnight and the standard 6-dose metyrapone tests for the differential diagnosis of ACTH-dependent Cushing's syndrome. DESIGN Retrospective pilot study based on all patients who completed both metyrapone tests and whose diagnoses were subsequently confirmed surgically. PATIENTS Sixty-three patients, 57 with pituitary tumours and 6 with ectopic ACTH syndrome, were studied. MEASUREMENTS The sensitivity and specificity were determined using stimulation of plasma 11-deoxycortisol and suppression of plasma cortisol as endpoints for the overnight test and stimulation of urine 17-hydroxysteroid and plasma 11-deoxycortisol as endpoints for the standard test. The test response that gave the highest sensitivity for the diagnosis of Cushing's disease, with 100% specificity, was determined for each test endpoint. RESULTS Both the stimulation of plasma 11-deoxycortisol and the suppression of plasma cortisol were expressed as the ratio of the post-metyrapone value to baseline. A plasma 11-deoxycortisol ratio > than 220 or a plasma cortisol ratio > 0.6 was associated with 100% specificity for diagnosis of Cushing's disease by the overnight test. When these two criteria were combined, the sensitivity was 65%, which was significantly higher than that obtained using either steroid alone: 42% for both plasma 11-deoxycortisol and plasma cortisol. The sensitivity of 65% for the overnight test, at 100% specificity, was nearly identical to that of the standard test in the same patients: 67% for the combined criterion of > 70% stimulation of urine 17-hydroxysteroid or > 480-fold increase of plasma 11-deoxycortisol. When the criteria for both the overnight and standard tests were combined, the sensitivity increased to 84%, which was higher than that obtained using the criteria for either test alone (P < 0.02). CONCLUSION The single-dose overnight metyrapone test, which can be performed in 24 hours and which avoids the disadvantages associated with timed urine collections, has a nearly identical sensitivity (for a specificity of 100%) as the 6-dose standard metyrapone test for the diagnosis of Cushing's disease. Furthermore, the diagnostic performance of a criterion that combines the results of both tests is significantly better than the best criterion for either test alone. C1 NICHHD, DEV ENDOCRINOL BRANCH, NIH, BETHESDA, MD 20892 USA. NINCDS, SURG NEUROL BRANCH, NIH, BETHESDA, MD 20892 USA. NR 28 TC 7 Z9 7 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA OSNEY MEAD, OXFORD, OXON, ENGLAND OX2 0EL SN 0300-0664 J9 CLIN ENDOCRINOL JI Clin. Endocrinol. PD OCT PY 1996 VL 45 IS 4 BP 483 EP 491 DI 10.1046/j.1365-2265.1996.8170827.x PG 9 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VU372 UT WOS:A1996VU37200019 PM 8959090 ER PT J AU Li, Q Sun, B Dastgheib, K Chan, CC AF Li, Q Sun, B Dastgheib, K Chan, CC TI Suppressive effect of transforming growth factor beta 1 on the recurrence of experimental melanin protein-induced uveitis: Upregulation of ocular interleukin-10 SO CLINICAL IMMUNOLOGY AND IMMUNOPATHOLOGY LA English DT Article ID AUTOIMMUNE ANTERIOR UVEITIS; EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS; RETINAL-PIGMENT EPITHELIUM; TGF-BETA; MESSENGER-RNA; CILIARY BODY; LATENT FORMS; T-CELLS; INDUCTION; ANTIGEN AB Uveitis is induced in Lewis rats by immunization with bovine melanin protein (BMP) derived from the uvea and retinal pigment epithelium. Recurrence of this experimental melanin protein-induced uveitis (EMIU) develops after footpad injection of a minimal amount of Salmonella typhimurium endotoxin (LPS) following the remission of EMIU. To investigate the effect of transforming growth factor beta 1 (TGF beta 1) on the recurrence of EMIU, 5 mu g LPS booster was given to Lewis rats by footpad injection on Day 45 after BMP immunization. Daily TGF beta 1 or phosphate-buffered saline was administered either from Day 0 to 7 (group 1) or from Day 7 to 13 (group 2) after LPS booster. Delayed-type hypersensitivity (DTH) ear test was conducted on Day 12 after LPS booster and eye and blood were collected on Day 14. The incidence and severity of recurrent uveitis markedly decreased in both groups of TGF beta 1-treated rats. A lower level of serum BMP antibody was also observed by agglutination in these groups. There was no statistical difference in DTH responses between the treated and control groups. Ocular cytokine mRNA of group 1 and controls was analyzed by RT-PCR, Interleukin (IL)-2 and interferon-gamma were not detectable. IL-4 was identified at a similar level in both groups. A higher level of IL-10 was observed in group 1 rats, We conclude that TGF beta 1 suppresses recurrent EMIU, probably through upregulation of IL-10. (C) 1996 Academic Press, Inc. RP Li, Q (reprint author), NEI, IMMUNOL LAB, BLDG 10, BETHESDA, MD 20892 USA. NR 33 TC 15 Z9 15 U1 0 U2 0 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0090-1229 J9 CLIN IMMUNOL IMMUNOP JI Clin. Immunol. Immunopathol. PD OCT PY 1996 VL 81 IS 1 BP 55 EP 61 DI 10.1006/clin.1996.0157 PG 7 WC Immunology; Pathology SC Immunology; Pathology GA VL236 UT WOS:A1996VL23600009 PM 8808642 ER PT J AU Beekmann, SE Fahrner, R Gerberding, JL Koziol, D Henderson, DK AF Beekmann, SE Fahrner, R Gerberding, JL Koziol, D Henderson, DK TI Risk factors for HIV seroconversion in a prospective study of zidovudine (ZDV) post-exposure prophylaxis (PEP). SO CLINICAL INFECTIOUS DISEASES LA English DT Meeting Abstract C1 NIH,CTR CLIN,BETHESDA,MD 20892. SAN FRANCISCO GEN HOSP,SAN FRANCISCO,CA 94110. UNIV IOWA,COLL MED,IOWA CITY,IA 52242. NR 0 TC 0 Z9 0 U1 0 U2 3 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 1058-4838 J9 CLIN INFECT DIS JI Clin. Infect. Dis. PD OCT PY 1996 VL 23 IS 4 BP 26 EP 26 PG 1 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA VN246 UT WOS:A1996VN24600074 ER PT J AU Kenney, R Sacks, D Gam, A AF Kenney, R Sacks, D Gam, A TI Interleukin-12 as an adjuvant for a vaccine against Leishmania amazonensis SO CLINICAL INFECTIOUS DISEASES LA English DT Meeting Abstract C1 US FDA,CTR BIOL EVALUAT & RES,BETHESDA,MD. NIAID,NIH,BETHESDA,MD 20892. NR 0 TC 0 Z9 0 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 1058-4838 J9 CLIN INFECT DIS JI Clin. Infect. Dis. PD OCT PY 1996 VL 23 IS 4 BP 64 EP 64 PG 1 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA VN246 UT WOS:A1996VN24600112 ER PT J AU Pappas, PG Hamill, RJ Kauffman, CA Bradsher, RW Mckinsey, DS Cloud, GW Dismukes, WE AF Pappas, PG Hamill, RJ Kauffman, CA Bradsher, RW Mckinsey, DS Cloud, GW Dismukes, WE TI Treatment of cryptococcal meningitis in non-HIV infected patients - A randomized comparative trial. SO CLINICAL INFECTIOUS DISEASES LA English DT Meeting Abstract C1 UNIV ALABAMA,BIRMINGHAM,AL. NIAID,BETHESDA,MD 20892. PFIZER CORP,NEW YORK,NY. NR 0 TC 0 Z9 0 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 1058-4838 J9 CLIN INFECT DIS JI Clin. Infect. Dis. PD OCT PY 1996 VL 23 IS 4 BP 73 EP 73 PG 1 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA VN246 UT WOS:A1996VN24600121 ER PT J AU Ohl, CA Koles, NL Guelde, G Witherspoon, KJ Mond, JJ Wahl, LM Pollack, M AF Ohl, CA Koles, NL Guelde, G Witherspoon, KJ Mond, JJ Wahl, LM Pollack, M TI Lipopolysaccharide (LPS)-specific monoclonal antibodies (MAbs) regulate LPS uptake and LPS-induced mitogenic responses by human peripheral blood B lymphocytes (PBL). SO CLINICAL INFECTIOUS DISEASES LA English DT Meeting Abstract C1 USUHS,BETHESDA,MD. NIDR,NIH,BETHESDA,MD 20892. NR 0 TC 0 Z9 0 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 1058-4838 J9 CLIN INFECT DIS JI Clin. Infect. Dis. PD OCT PY 1996 VL 23 IS 4 BP 87 EP 87 PG 1 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA VN246 UT WOS:A1996VN24600134 ER PT J AU Domachowske, JB Rosenberg, HF Dyer, KD Malech, HL AF Domachowske, JB Rosenberg, HF Dyer, KD Malech, HL TI Nitric oxide alters gene expression in murine RAW 264 cells. SO CLINICAL INFECTIOUS DISEASES LA English DT Meeting Abstract C1 NIAID,HOST DEF LAB,NIH,BETHESDA,MD 20892. NR 0 TC 0 Z9 0 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 1058-4838 J9 CLIN INFECT DIS JI Clin. Infect. Dis. PD OCT PY 1996 VL 23 IS 4 BP 94 EP 94 PG 1 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA VN246 UT WOS:A1996VN24600141 ER PT J AU Walsh, TJ Martinez, A Peter, J Unsworth, E Cuttitta, F AF Walsh, TJ Martinez, A Peter, J Unsworth, E Cuttitta, F TI Antimicrobial activity of adrenomedullin and its gene-related peptides. SO CLINICAL INFECTIOUS DISEASES LA English DT Meeting Abstract C1 NCI,NIH,BETHESDA,MD 20892. RI Martinez, Alfredo/A-3077-2013 OI Martinez, Alfredo/0000-0003-4882-4044 NR 0 TC 5 Z9 5 U1 0 U2 1 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 1058-4838 J9 CLIN INFECT DIS JI Clin. Infect. Dis. PD OCT PY 1996 VL 23 IS 4 BP 96 EP 96 PG 1 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA VN246 UT WOS:A1996VN24600143 ER PT J AU Piscitelli, SC Flexner, C Minor, JR Polis, MA Masur, H AF Piscitelli, SC Flexner, C Minor, JR Polis, MA Masur, H TI Drug interactions in patients infected with human immunodeficiency virus SO CLINICAL INFECTIOUS DISEASES LA English DT Article ID RIFABUTIN PROPHYLAXIS; KETOCONAZOLE; CLARITHROMYCIN; PHARMACOKINETICS; TERFENADINE; AIDS; BIOAVAILABILITY; FLUCONAZOLE; ZIDOVUDINE; DISEASE AB Patients with AIDS who are receiving optimal medical care, including combination therapy with antiretroviral agents and more effective prophylaxis and therapy for opportunistic infections and neoplasms, are surviving longer. However, the potential for drug interactions in these patients is increased because many of the currently used antibiotics and antiviral agents have profound effects on the hepatic cytochrome P-450 enzyme system, on renal tubular function, and on bone marrow function. In this AIDS Commentary, Dr. Piscitelli and colleagues have succinctly reviewed the current state of our knowledge regarding the potential for additive or synergistic drug interactions that can result in enhanced toxicity or, alternatively, augmented therapeutic benefit, Information on these interactions will become more important as more intensive and effective therapy becomes available for persons with far-advanced infection due to human immunodeficiency virus type 1. C1 NIH,WARREN G MAGNUSON CLIN CTR,DEPT PHARM,BETHESDA,MD 20892. NIH,WARREN G MAGNUSON CLIN CTR,DEPT CRIT CARE MED,BETHESDA,MD 20892. NIH,IMMUNOREGULAT LAB,BETHESDA,MD 20892. JOHNS HOPKINS UNIV,SCH MED,DIV CLIN PHARMACOL,BALTIMORE,MD. NR 58 TC 109 Z9 113 U1 0 U2 2 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 1058-4838 J9 CLIN INFECT DIS JI Clin. Infect. Dis. PD OCT PY 1996 VL 23 IS 4 BP 685 EP 693 PG 9 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA VN246 UT WOS:A1996VN24600002 PM 8909827 ER PT J AU Miller, KD Mican, JAM Davey, RT AF Miller, KD Mican, JAM Davey, RT TI Asymptomatic solitary pulmonary nodules due to Cryptococcus neoformans in patients infected with human immunodeficiency virus SO CLINICAL INFECTIOUS DISEASES LA English DT Article ID AIDS; MANIFESTATIONS; DISEASE; UTILITY; ANTIGEN AB We report the cases of three HIV-positive patients with solitary pulmonary nodules caused by Cryptococcus neoformans. Although human infection with C. neoformans occurs via the respiratory tract, isolated pulmonary infection in HIV-positive patients, in contrast with HIV-negative patients, has been thought to be relatively rare. When isolated pulmonary disease in HIV-infested patients has been described, most of the patients have been symptomatic (symptoms have included fever, cough, and dyspnea). In addition, these patients have had diffuse interstitial infiltrates, alveolar infiltrates, or nodular infiltrates that have often been associated with hilar adenopathy and occasionally with pleural effusions. None of the patients in the previously reported series have had lesions described as small, asymptomatic, isolated pulmonary nodules. C1 NIAID,NIH,IMMUNOREGULAT LAB,BETHESDA,MD 20892. NR 17 TC 17 Z9 17 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 1058-4838 J9 CLIN INFECT DIS JI Clin. Infect. Dis. PD OCT PY 1996 VL 23 IS 4 BP 810 EP 812 PG 3 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA VN246 UT WOS:A1996VN24600024 PM 8909849 ER PT J AU Lucey, DR Clerici, M Shearer, GM AF Lucey, DR Clerici, M Shearer, GM TI Type 1 and type 2 cytokine dysregulation in human infectious, neoplastic, and inflammatory diseases SO CLINICAL MICROBIOLOGY REVIEWS LA English DT Review ID HUMAN-IMMUNODEFICIENCY-VIRUS; T-CELL CLONES; TUMOR-NECROSIS-FACTOR; RESPIRATORY SYNCYTIAL VIRUS; INTERFERON-GAMMA PRODUCTION; MESSENGER-RNA EXPRESSION; SYSTEMIC LUPUS-ERYTHEMATOSUS; COLONY-STIMULATING FACTOR; BLOOD MONONUCLEAR-CELLS; DELAYED-TYPE HYPERSENSITIVITY AB In the mid-1980s, Mosmann, Coffman, and their colleagues discovered that murine CD4(+) helper T-cell clones could be distinguished by the cytokines they synthesized. The isolation of human Th1 and Th2 clones by Romagnani and coworkers in the early 1990s has led to a large number of reports on the effects of Th1 and Th2 on the human immune system. More recently, cells other than CD4(+) T cells, including CD8(+) T cells, monocytes NK cells, B cells, eosinophils, mast cells, basophils, and other cells, have been shown to be capable of producing ''Th1'' and ''Th2'' cytokines. In this review, we examine the literature on human diseases, using the nomenclature of type 1 (Th1-like) and type 2 (Th2-like) cytokines, which includes all cell types producing these cytokines rather than only CD4(+) T cells. Type I cytokines include interleukin-2 (IL-2), gamma interferon, IL-12 and tumor necrosis factor beta, while type 2 cytokines include IL-4, IL-5, IL-6, IL-10, and IL-13. In general, type 1 cytokines favor the development of a strong cellular immune response whereas type 2 cytokines favor a strong humoral immune response. Some of these type 1 and type 2 cytokines are cross-regulatory. For example, gamma interferon on and IL-12 decrease the levels of type 2 cytokines whereas IL-4 and IL-10 decrease the levels of type 1 cytokines. We use this cytokine perspective to examine human diseases including infections due to viruses, bacteria, parasites, and fungi, as well as selected neoplastic, atopic, rheumatologic, autoimmune, and idiopathic-inflammatory conditions. Clinically, type 1 cytokine-predominant responses should be suspected in any delayed-type hypersensitivity-like sensitivity-like granulomatous reactions and in infections with intracellular pathogens, whereas conditions involving hypergammaglobulinemia, increased immunoglobulin E levels, and/or eosinophilia are suggestive of type 2 cytokine-predominant conditions. If this immunologic concept is relevant to human diseases the potential exists for novel cytokine-based therapies and novel cytokine-directed preventive vaccines for such diseases. C1 NCI, EXPT IMMUNOL BRANCH, BETHESDA, MD 20892 USA. RI Liu, Xia/L-9425-2013 NR 431 TC 446 Z9 468 U1 1 U2 7 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0893-8512 J9 CLIN MICROBIOL REV JI Clin. Microbiol. Rev. PD OCT PY 1996 VL 9 IS 4 BP 532 EP + PG 0 WC Microbiology SC Microbiology GA VL539 UT WOS:A1996VL53900007 PM 8894351 ER PT J AU Prell, GD Green, JP Khandelwal, JK Wyatt, RJ Lawson, WB Jaeger, AC Kaufmann, CA Kirch, DG AF Prell, GD Green, JP Khandelwal, JK Wyatt, RJ Lawson, WB Jaeger, AC Kaufmann, CA Kirch, DG TI pros-Methylimidazoleacetic acid in cerebrospinal fluid of patients with chronic schizophrenia: Relationships to ratings of symptoms, ventricular brain ratios, and rates of urine excretion SO CLINICAL NEUROPHARMACOLOGY LA English DT Review DE pros-methylimidazoleacetic acid; cerebrospinal fluid; schizophrenia; ventricular brain ratio; positive symptoms; glutamate ID HISTAMINE METABOLITES; CARNOSINE; MICE; CSF AB Concentrations of pros-methylimidazoleacetic acid (p-MIAA) were measured in cerebrospinal fluid of 30 patients with chronic schizophrenia. Levels of p-MIAA correlated negatively with mean scores of the Psychiatric Symptom Assessment Scale for positive symptoms (r = -0.48), but not negative symptoms, and with ventricular brain ratios (r = -0.48). Patients with abnormal ventricular enlargements had much lower concentrations of p-MIAA than those with normal ventricles, These results suggest that processes that reduce accumulation of p-MIAA in CSF may be associated with increased severity of symptoms among patients with chronic schizophrenia. C1 ST ELIZABETH HOSP,NEUROPSYCHIAT BRANCH,NIMH,WASHINGTON,DC. RP Prell, GD (reprint author), CUNY,MT SINAI SCH MED,DEPT PHARMACOL,BOX 1215,1 GUSTAVE L LEVY PL,NEW YORK,NY 10029, USA. OI Lawson, William/0000-0002-9324-7090 FU NIMH NIH HHS [MH-31805]; NINDS NIH HHS [NS-28012] NR 22 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT-RAVEN PUBL PI PHILADELPHIA PA 227 EAST WASHINGTON SQ, PHILADELPHIA, PA 19106 SN 0362-5664 J9 CLIN NEUROPHARMACOL JI Clin. Neuropharmacol. PD OCT PY 1996 VL 19 IS 5 BP 415 EP 419 DI 10.1097/00002826-199619050-00004 PG 5 WC Clinical Neurology; Pharmacology & Pharmacy SC Neurosciences & Neurology; Pharmacology & Pharmacy GA VH263 UT WOS:A1996VH26300004 PM 8889284 ER PT J AU Parke, RD Ornstein, P Reiser, J ZahnWaxler, C AF Parke, RD Ornstein, P Reiser, J ZahnWaxler, C TI Meacham misses the mark SO CONTEMPORARY PSYCHOLOGY LA English DT Letter C1 UNIV N CAROLINA,CHAPEL HILL,NC. NIMH,ROCKVILLE,MD 20857. RP Parke, RD (reprint author), UNIV CALIF RIVERSIDE,RIVERSIDE,CA 92521, USA. NR 1 TC 0 Z9 0 U1 0 U2 0 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 SN 0010-7549 J9 CONTEMP PSYCHOL JI Comtemp. Psychol. PD OCT PY 1996 VL 41 IS 10 BP 1071 EP 1072 PG 2 WC Psychology, Multidisciplinary SC Psychology GA VN267 UT WOS:A1996VN26700067 ER PT J AU Levine, RJ Esterlitz, JR Raymond, EG DerSimonian, R Hauth, JC BenCuret, L Sibai, BM Catalano, PM Morris, CD Clemens, JD Ewell, MG Friedman, SA Goldenberg, RL Jacobson, SL Joffe, GM Klebanoff, MA Petrulis, AS RigauPerez, JG AF Levine, RJ Esterlitz, JR Raymond, EG DerSimonian, R Hauth, JC BenCuret, L Sibai, BM Catalano, PM Morris, CD Clemens, JD Ewell, MG Friedman, SA Goldenberg, RL Jacobson, SL Joffe, GM Klebanoff, MA Petrulis, AS RigauPerez, JG TI Trial of calcium for preeclampsia prevention (CPEP): Rationale, design, and methods SO CONTROLLED CLINICAL TRIALS LA English DT Article DE clinical trial; calcium; hypertension; proteinuria; preeclampsia; pregnancy ID PREGNANCY-INDUCED HYPERTENSION; SAMPLE-SIZE; SUPPLEMENTATION; POWER; RISK AB The results of ten clinical trials suggest that supplemental calcium may prevent preeclampsia. However, methodologic problems and differences in study design limit the acceptance of the results and their relevance to other patient populations. Many of the trials were conducted in countries where, unlike the United States, the usual daily diet contained little calcium. Moreover, none of the trials has reported the outcome of systematic surveillance for urolithiasis, a potential complication of calcium supplementation. In response to the need for a thorough evaluation of the effects of calcium supplementation for the prevention of preeclampsia in the United States, the trial of Calcium for Preeclampsia Prevention (CPEP) was undertaken at five university medical centers. Healthy nulliparous patients were randomly assigned to receive either 2 g supplemental calcium daily (n = 2295) or placebo (n = 2294) in a double-blind study. Study tablets were administered beginning from 13 to 21 completed weeks of gestation and continued until the termination of pregnancy. CPEP employed detailed diagnostic criteria, standardized techniques of measurement, and systematic surveillance for the major study endpoints and for urolithiasis. The nutrient intake of each patient was assessed at randomization and at 32-33 weeks gestation. This report describes the study rationale, design, and methods. (C) Elsevier Science Inc. C1 EMMES CORP,POTOMAC,MD. UNIV ALABAMA,DEPT OBSTET & GYNECOL,BIRMINGHAM,AL 35294. UNIV NEW MEXICO,DEPT OBSTET & GYNECOL,ALBUQUERQUE,NM 87131. UNIV TENNESSEE,DEPT OBSTET & GYNECOL,MEMPHIS,TN 38103. CASE WESTERN RESERVE UNIV,METROHLTH MED CTR,DEPT REPROD BIOL,CLEVELAND,OH. CASE WESTERN RESERVE UNIV,METROHLTH MED CTR,DEPT NEPHROL,CLEVELAND,OH. OREGON HLTH SCI UNIV,DEPT MED,PORTLAND,OR 97201. OREGON HLTH SCI UNIV,DEPT OBSTET & GYNECOL,PORTLAND,OR 97201. RP Levine, RJ (reprint author), NICHHD,DIV EPIDEMIOL STAT & PREVENT RES,BLDG 6100,ROOM 7B03,BETHESDA,MD 20892, USA. FU NICHD NIH HHS [N01-HD-1-3121, N01-HD-1-3122, N01-HD-1-3123] NR 35 TC 54 Z9 56 U1 0 U2 4 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 SN 0197-2456 J9 CONTROL CLIN TRIALS JI Controlled Clin. Trials PD OCT PY 1996 VL 17 IS 5 BP 442 EP 469 DI 10.1016/S0197-2456(96)00106-7 PG 28 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA VR948 UT WOS:A1996VR94800007 PM 8932976 ER PT J AU Schreuders, PD Smith, ED Cole, KW Valencia, MDP Laughinghouse, A Mazur, P AF Schreuders, PD Smith, ED Cole, KW Valencia, MDP Laughinghouse, A Mazur, P TI Characterization of intraembryonic freezing in Anopheles gambiae embryos SO CRYOBIOLOGY LA English DT Article ID DROSOPHILA EMBRYOS; MOUSE EMBRYOS; CRYOPRESERVATION; VITRIFICATION AB Intraembryonic freezing (IEF) in Anopheles mosquito embryos has been evaluated by differential scanning calorimetry with respect to embryo age, temperature, rate and duration of cooling, and absence or presence of extraembryonic ice. The initial temperatures for intraembryonic ice nucleation were -30.1 +/- 0.3, -28.4 +/- 0.4, and -29.1 +/- 0.2 degrees C for embryos incubated for 15 h at 17 degrees C, 15 h at 26 degrees C, and 24 h at 26 degrees C, respectively, after oviposition. The first value is slightly but significantly lower than the latter two. These values were obtained on embryos in which the surface water was removed by brief drying; however, the values were nearly identical when external water and ice were present. Not only were the embryos of all three ages able to supercool at least transiently to -26 degrees C, but they could remain supercooled for up to 4 h at -20 degrees C after being cooled to -20 degrees C at 10 degrees C/min or (in the case of embryos incubated for 15 h at 26 degrees C) at 100 degrees C/min. The amount of freezable water in single embryos has been calculated from the differential scanning calorimetry measurements to be 3.45 +/- 0.08, 3.46 +/- 0.08, and 3.53 +/- 0.06 mu g for embryos incubated for 15 h at 17 degrees C, 15 h at 26 degrees C, and 24 h at 26 degrees C, respectively. The differences are not significant. The corresponding values for the total water contents for embryos of the three ages were 4.04 +/- 0.20, 3.72 +/- 0.16, and 3.98 +/- 0.10 mu g, values that also did not differ significantly. Water thus makes up similar to 74% of the total weight of the embryo (similar to 5.3 mu g) and about 91% of that water is freezable. Total water contents were determined gravimetrically after extensive air and vacuum drying. The kinetics of dehydration were determined during the air drying. They differed substantially among the three ages. The embryos incubated for 15 h at 17 degrees C lost water at about four times the rate of those incubated for 15 h at 26 degrees C and 10 times the rate of the embryos incubated for 24 h at 26 degrees C. (C) 1996 Academic Press, Inc. C1 OAK RIDGE NATL LAB,DIV BIOL,FUNDAMENTAL & APPL CRYOBIOL GRP,OAK RIDGE,TN 37831. NIH,PARASIT DIS LAB,BETHESDA,MD 20892. RP Schreuders, PD (reprint author), UNIV TENNESSEE,OAK RIDGE GRAD SCH BIOMED SCI,KNOXVILLE,TN, USA. FU NIAID NIH HHS [1RO1AI3624301A1] NR 28 TC 8 Z9 8 U1 0 U2 0 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0011-2240 J9 CRYOBIOLOGY JI Cryobiology PD OCT PY 1996 VL 33 IS 5 BP 487 EP 501 DI 10.1006/cryo.1996.9999 PG 15 WC Biology; Physiology SC Life Sciences & Biomedicine - Other Topics; Physiology GA VQ074 UT WOS:A1996VQ07400001 PM 8893508 ER PT J AU Zhi, L Karesh, WB Janczewski, DN FrazierTaylor, H Sajuthi, D Gombek, F Andau, M Martenson, JS OBrien, SJ AF Zhi, L Karesh, WB Janczewski, DN FrazierTaylor, H Sajuthi, D Gombek, F Andau, M Martenson, JS OBrien, SJ TI Genomic differentiation among natural populations of orang-utan (Pongo pygmaeus) SO CURRENT BIOLOGY LA English DT Article ID MITOCHONDRIAL-DNA; RESTRICTION ENDONUCLEASES; HOMINOID PHYLOGENY; FINGERPRINTS; EVOLUTION; TREES; APES AB Background: Orang-utans exist today in small isolated populations on the islands of Borneo (subspecies Pongo pygmaeus pygmaeus) and Sumatra (subspecies P. p. abelii). Although, on the basis of their morphological, behavioral and cytogenetical characteristics, the Bornean and Sumatran orang-utan populations are generally considered as two separate subspecies, there is no universal agreement as to whether their genetic differentiation is sufficient to consider and manage them as species, subspecies or population level taxonomic units. A more precise phylogenetic description would affect many conservation management decisions about captive and free-ranging orang-utans. Results: We analyzed the amount and patterns of molecular genetic variation in orang-utan populations using cellular DNA from orang-utans from two locations in Sumatra and nine locations - representing four isolated populations - in Borneo, Genetic and phylogenetic analyses of mitochondrial DNA restriction fragment length polymorphisms, nuclear minisatellite (or variable number tandem repeat) loci and mitochondrial 16S ribosomal RNA sequences led to three major findings. First, the genetic distance and phylogenetic differentiation between Sumatran and Bornean orang-utans is large, greater than that between the common chimpanzee, Pan troglodytes, and the pygmy chimpanzee or bonobo, Pan paniscus. The genetic distance suggests that the two island subspecies diverged similar to 1.5-1.7 million years ago, well before the two islands separated and long enough for species-level differentiation. Second, there is considerable endemic genetic diversity within the Bornean and Sumatran orang-utan populations, suggesting that they have not experienced recent bottlenecks or founder effects. And third, there is little genetic differentiation among four geographically isolated populations of Bornean orang-utans, consistent with gene flow having occurred between them until recently. Conclusions: Our results are consistent with the view that the genetic differentiation between Sumatran and Bornean orang-utans has reached the level of distinct species, Furthermore, our findings indicate that there is not a genetic imperative for the separate management of geographically isolated Bornean populations. (C) Current Biology Ltd ISSN 0960-9822 C1 NCI,LAB GENOM DIVERS,FREDERICK,MD 21702. NEW YORK ZOOL SOC,BRONX,NY 10460. WOODLAND PK ZOO,SEATTLE,WA 98103. INST PERTANIAN BOGOR,BOGOR,INDONESIA. NATL PARKS & WILDLIFE DIV,KUCHING,SARAWAK,MALAYSIA. DEPT WILDLIFE,KOTA KINABALU,SABAH,MALAYSIA. NR 55 TC 53 Z9 55 U1 2 U2 22 PU CURRENT BIOLOGY LTD PI LONDON PA 34-42 CLEVELAND STREET, LONDON, ENGLAND W1P 6LB SN 0960-9822 J9 CURR BIOL JI Curr. Biol. PD OCT 1 PY 1996 VL 6 IS 10 BP 1326 EP 1336 DI 10.1016/S0960-9822(02)70719-7 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA VN635 UT WOS:A1996VN63500029 PM 8939569 ER PT J AU Sandkvist, M Bagdasarian, M AF Sandkvist, M Bagdasarian, M TI Secretion of recombinant proteins by gram-negative bacteria SO CURRENT OPINION IN BIOTECHNOLOGY LA English DT Article ID DISULFIDE BOND FORMATION; ESCHERICHIA-COLI; ERWINIA-CHRYSANTHEMI; VIBRIO-CHOLERAE; HETEROLOGOUS PROTEINS; EXTRACELLULAR SECRETION; PERIPLASMIC PROTEIN; BETA-GALACTOSIDASE; ABC-TRANSPORTERS; GENE-EXPRESSION AB During the past few years, significant progress has been made towards our understanding of the molecular mechanisms governing the translocation of proteins through bacterial cell membranes. Successful attempts in promoting the secretion of recombinant proteins by employing this knowledge and by empirical efforts have been registered. However, a further in-depth understanding of membrane-translocation mechanisms is required before predictable manipulations of secretion systems can be made to secrete native recombinant proteins that are not naturally targeted to the extracellular compartment. C1 MICHIGAN STATE UNIV, E LANSING, MI 48824 USA. RP NIDR, NATL INST HLTH, BLDG 30, ROOM 313, BETHESDA, MD 20892 USA. NR 61 TC 28 Z9 30 U1 0 U2 4 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0958-1669 EI 1879-0429 J9 CURR OPIN BIOTECH JI Curr. Opin. Biotechnol. PD OCT PY 1996 VL 7 IS 5 BP 505 EP 511 DI 10.1016/S0958-1669(96)80053-X PG 7 WC Biochemical Research Methods; Biotechnology & Applied Microbiology SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology GA VL302 UT WOS:A1996VL30200005 PM 8939628 ER PT J AU Makalowski, W Recipon, H Baxevanis, AD AF Makalowski, W Recipon, H Baxevanis, AD TI Web alert - Expression systems SO CURRENT OPINION IN BIOTECHNOLOGY LA English DT Article C1 NIH,NATL CTR HUMAN GENOME RES,BETHESDA,MD 20892. RP Makalowski, W (reprint author), NIH,NATL CTR BIOTECHNOL INFORMAT,BLDG 38A,ROOM 8N-805,BETHESDA,MD 20894, USA. RI Makalowski, Wojciech/I-2843-2016 NR 0 TC 0 Z9 0 U1 0 U2 0 PU CURRENT BIOLOGY LTD PI LONDON PA 34-42 CLEVELAND STREET, LONDON, ENGLAND W1P 6LB SN 0958-1669 J9 CURR OPIN BIOTECH JI Curr. Opin. Biotechnol. PD OCT PY 1996 VL 7 IS 5 BP 563 EP 563 DI 10.1016/S0958-1669(96)80063-2 PG 1 WC Biochemical Research Methods; Biotechnology & Applied Microbiology SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology GA VL302 UT WOS:A1996VL30200015 PM 8939633 ER PT J AU Dawid, IB Brown, DD AF Dawid, IB Brown, DD TI Differentiation from bacteria to humans - Editorial overview SO CURRENT OPINION IN GENETICS & DEVELOPMENT LA English DT Editorial Material C1 CARNEGIE INST WASHINGTON,DEPT EMBRYOL,BALTIMORE,MD 21210. RP Dawid, IB (reprint author), NICHHD,GENET MOL LAB,BETHESDA,MD 20892, USA. NR 0 TC 2 Z9 2 U1 0 U2 0 PU CURRENT BIOLOGY LTD PI LONDON PA 34-42 CLEVELAND STREET, LONDON, ENGLAND W1P 6LB SN 0959-437X J9 CURR OPIN GENET DEV JI Curr. Opin. Genet. Dev. PD OCT PY 1996 VL 6 IS 5 BP 523 EP 525 DI 10.1016/S0959-437X(96)80078-5 PG 3 WC Cell Biology; Genetics & Heredity SC Cell Biology; Genetics & Heredity GA VP615 UT WOS:A1996VP61500001 PM 8939731 ER PT J AU Minucci, S Ozato, K AF Minucci, S Ozato, K TI Retinoid receptors in transcriptional regulation SO CURRENT OPINION IN GENETICS & DEVELOPMENT LA English DT Article ID THYROID-HORMONE RECEPTOR; LIGAND-DEPENDENT ACTIVATION; NUCLEAR RECEPTOR; X RECEPTOR; ACID RECEPTORS; CONFORMATIONAL-CHANGES; ESTROGEN-RECEPTOR; CRYSTAL-STRUCTURE; RXR HETERODIMERS; BINDING DOMAIN AB Our understanding of the mechanism of action of retinoids has been greatly expanded by a series of recent findings. First, the three-dimensional structure of the ligand-binding domain of two retinoid receptors has been solved and suggests that ligand binding induces marked allosteric changes. Second, several co-factors interacting with the receptors have been cloned, some of which are capable of regulating the function of receptors. Third, the advent of synthetic retinoids helped define the activities of the receptors. Fourth, the study of the in vivo receptor-DNA interactions has revealed a previously unrecognized role of the ligand in regulating the stability of receptor-DNA complexes. These advances have revealed complex molecular interactions operating at multiple levels, opening new avenues of research for addressing their mechanisms. RP Minucci, S (reprint author), NICHHD,NIH,LAB MOL GROWTH REGULAT,9000 ROCKVILLE PIKE,BETHESDA,MD 20892, USA. RI Minucci, Saverio/J-9669-2012 NR 67 TC 61 Z9 62 U1 0 U2 1 PU CURRENT BIOLOGY LTD PI LONDON PA 34-42 CLEVELAND STREET, LONDON, ENGLAND W1P 6LB SN 0959-437X J9 CURR OPIN GENET DEV JI Curr. Opin. Genet. Dev. PD OCT PY 1996 VL 6 IS 5 BP 567 EP 574 DI 10.1016/S0959-437X(96)80085-2 PG 8 WC Cell Biology; Genetics & Heredity SC Cell Biology; Genetics & Heredity GA VP615 UT WOS:A1996VP61500008 PM 8939720 ER PT J AU Gottesman, MM Pastan, I Ambudkar, SV AF Gottesman, MM Pastan, I Ambudkar, SV TI P-glycoprotein and multidrug resistance SO CURRENT OPINION IN GENETICS & DEVELOPMENT LA English DT Article ID PROTEIN-KINASE-C; BINDING-CASSETTE TRANSPORTER; MUTATIONAL ANALYSIS; TISSUE DISTRIBUTION; STEROID-HORMONES; GENE-EXPRESSION; ATPASE ACTIVITY; DRUG TRANSPORT; LABELING SITES; LINKER REGION AB Although the phenomenon of simultaneous resistance to multiple cytotoxic drugs (multidrug resistance) in cancer cells has been discussed for more than two decades, and the human and mouse genes encoding an energy-dependent transporter (the multidrug transporter or P-glycoprotein) responsible for multidrug resistance were cloned 10 years ago, there is still considerable controversy about the mechanism of action of this efflux pump and its true biological function. This review summarizes the current research on the mechanism of action of the multidrug transporter, including the hydrophobic cleaner and altered partitioning models, the possible function of P-glycoprotein as a chloride and/or ATP channel, the role of phosphorylation in its function and fact and speculation about its physiological role. C1 NCI,NIH,DIV BASIC SCI,MOL BIOL LAB,BETHESDA,MD 20892. RP Gottesman, MM (reprint author), NCI,NIH,DIV BASIC SCI,CELL BIOL LAB,37 CONVENT DR MSC,BETHESDA,MD 20892, USA. RI Ambudkar, Suresh/B-5964-2008 NR 65 TC 432 Z9 440 U1 2 U2 27 PU CURRENT BIOLOGY LTD PI LONDON PA 34-42 CLEVELAND STREET, LONDON, ENGLAND W1P 6LB SN 0959-437X J9 CURR OPIN GENET DEV JI Curr. Opin. Genet. Dev. PD OCT PY 1996 VL 6 IS 5 BP 610 EP 617 DI 10.1016/S0959-437X(96)80091-8 PG 8 WC Cell Biology; Genetics & Heredity SC Cell Biology; Genetics & Heredity GA VP615 UT WOS:A1996VP61500014 PM 8939727 ER PT J AU Rosenberg, SA AF Rosenberg, SA TI Secrecy in research: Cooperation and costimulation in immunology SO CURRENT OPINION IN IMMUNOLOGY LA English DT Editorial Material RP Rosenberg, SA (reprint author), NCI,SURG BRANCH,NIH,BETHESDA,MD 20892, USA. NR 3 TC 0 Z9 0 U1 0 U2 0 PU CURRENT BIOLOGY LTD PI LONDON PA 34-42 CLEVELAND STREET, LONDON, ENGLAND W1P 6LB SN 0952-7915 J9 CURR OPIN IMMUNOL JI Curr. Opin. Immunol. PD OCT PY 1996 VL 8 IS 5 BP 611 EP 612 DI 10.1016/S0952-7915(96)80074-4 PG 2 WC Immunology SC Immunology GA VL883 UT WOS:A1996VL88300001 PM 8902384 ER PT J AU OBrien, SJ Goedert, JJ AF OBrien, SJ Goedert, JJ TI HIV causes AIDS: Koch's postulates fulfilled SO CURRENT OPINION IN IMMUNOLOGY LA English DT Editorial Material ID HUMAN-IMMUNODEFICIENCY-VIRUS; PBL-SCID MICE; FLORIDA DENTIST; NEF GENE; INFECTION; TRANSMISSION; DISEASE; TYPE-1; RISK; PATHOGENICITY C1 NCI,ROCKVILLE,MD 20852. RP OBrien, SJ (reprint author), NCI,FREDERICK,MD 21702, USA. NR 85 TC 16 Z9 17 U1 2 U2 15 PU CURRENT BIOLOGY LTD PI LONDON PA 34-42 CLEVELAND STREET, LONDON, ENGLAND W1P 6LB SN 0952-7915 J9 CURR OPIN IMMUNOL JI Curr. Opin. Immunol. PD OCT PY 1996 VL 8 IS 5 BP 613 EP 618 DI 10.1016/S0952-7915(96)80075-6 PG 6 WC Immunology SC Immunology GA VL883 UT WOS:A1996VL88300002 PM 8902385 ER PT J AU Robbins, PF Kawakami, Y AF Robbins, PF Kawakami, Y TI Human tumor antigens recognized by T cells SO CURRENT OPINION IN IMMUNOLOGY LA English DT Review ID CYTOLYTIC LYMPHOCYTES-T; HUMAN GENE MAGE-3; HUMAN-MELANOMA; INFILTRATING LYMPHOCYTES; IN-VITRO; SYNTHETIC PEPTIDE; HLA-A2 MELANOMAS; PERIPHERAL-BLOOD; IMMUNODOMINANT PEPTIDE; MULTIPLE EPITOPES AB The ability of tumor-reactive T cells to mediate in vivo tumor regression has been demonstrated in murine tumor models and by the clinical responses to adoptive immunotherapy with tumor-infiltrating lymphocytes isolated from human melanomas. Investigations carried out in the past several years have resulted in the isolation of a number of the genes encoding antigens recognized by melanoma-reactive T cells. The ability of these products to serve as tumor regression antigens has now begun to be evaluated in clinical vaccine trials. RP Robbins, PF (reprint author), NCI,SURG BRANCH,DIV CLIN SCI,NIH,BLDG 10,ROOM 2B42,10 CTR DR,BETHESDA,MD 20892, USA. RI Kawakami, Yutaka /E-7429-2013 OI Kawakami, Yutaka /0000-0003-4836-2855 NR 81 TC 168 Z9 175 U1 0 U2 3 PU CURRENT BIOLOGY LTD PI LONDON PA 34-42 CLEVELAND STREET, LONDON, ENGLAND W1P 6LB SN 0952-7915 J9 CURR OPIN IMMUNOL JI Curr. Opin. Immunol. PD OCT PY 1996 VL 8 IS 5 BP 628 EP 636 DI 10.1016/S0952-7915(96)80078-1 PG 9 WC Immunology SC Immunology GA VL883 UT WOS:A1996VL88300005 PM 8902387 ER PT J AU Restifo, NP AF Restifo, NP TI The new vaccines: Building viruses that elicit antitumor immunity SO CURRENT OPINION IN IMMUNOLOGY LA English DT Review ID TUMOR-ASSOCIATED ANTIGEN; T-CELL RESPONSES; IN-VIVO; CARCINOEMBRYONIC ANTIGEN; CTL; IMMUNIZATION; LYMPHOCYTES; CANCER; METASTASES; APOPTOSIS AB Whereas cancer cells are poor immunogens, some viruses are capable of eliciting powerful and lifelong immunity. Recombinant viruses and plasmid DNA encoding tumor-associated antigens can elicit powerful and specific immune responses that can be enhanced by the use of cytokines and costimulatory molecules. These immune responses have destroyed growing tumor cells in experimental animal models. For the first time, immunotherapeutic strategies that employ recombinant viruses are being tested in clinical trials with cancer patients. RP Restifo, NP (reprint author), NCI,SURG BRANCH,NIH,BLDG 10,ROOM 2842,10 CTR DR,BETHESDA,MD 20892, USA. RI Restifo, Nicholas/A-5713-2008; OI Restifo, Nicholas P./0000-0003-4229-4580 FU Intramural NIH HHS [Z01 BC010763-01, Z99 CA999999] NR 54 TC 58 Z9 58 U1 0 U2 0 PU CURRENT BIOLOGY LTD PI LONDON PA 34-42 CLEVELAND STREET, LONDON, ENGLAND W1P 6LB SN 0952-7915 J9 CURR OPIN IMMUNOL JI Curr. Opin. Immunol. PD OCT PY 1996 VL 8 IS 5 BP 658 EP 663 DI 10.1016/S0952-7915(96)80082-3 PG 6 WC Immunology SC Immunology GA VL883 UT WOS:A1996VL88300009 PM 8902391 ER PT J AU Brustle, O McKay, RDG AF Brustle, O McKay, RDG TI Neuronal progenitors as tools for cell replacement in the nervous system SO CURRENT OPINION IN NEUROBIOLOGY LA English DT Article ID REGION-SPECIFIC DIFFERENTIATION; ADULT MAMMALIAN FOREBRAIN; BILATERAL 6-OHDA LESIONS; FIBROBLAST GROWTH-FACTOR; EMBRYONIC STEM-CELLS; HIPPOCAMPAL-NEURONS; MOUSE-BRAIN; DOPAMINERGIC MICROTRANSPLANTS; NIGROSTRIATAL PATHWAY; NEURAL PROGENITORS AB The clinical prospect of using neural precursor cells for reconstructive approaches in the nervous system has received strong impetus from a recent series of important experimental findings. Transplantation studies in the developing brain have demonstrated that migration and differentiation of neural precursor cells are regulated predominantly by environmental signals, Several observations suggest that the mature CNS retains at least some of these guidance cues. These findings, together with recent evidence for the persistence of neural stem cells in the adult mammalian brain, have made precursor cell recruitment a new focus in CNS reconstruction. RP Brustle, O (reprint author), NINCDS,MOL BIOL LAB,NIH,BETHESDA,MD 20892, USA. NR 81 TC 101 Z9 103 U1 8 U2 10 PU CURRENT BIOLOGY LTD PI LONDON PA 34-42 CLEVELAND STREET, LONDON, ENGLAND W1P 6LB SN 0959-4388 J9 CURR OPIN NEUROBIOL JI Curr. Opin. Neurobiol. PD OCT PY 1996 VL 6 IS 5 BP 688 EP 695 DI 10.1016/S0959-4388(96)80104-8 PG 8 WC Neurosciences SC Neurosciences & Neurology GA VR586 UT WOS:A1996VR58600018 PM 8937835 ER PT J AU Moore, DH Epstein, L Reeder, J Wheeless, L Waldman, FM Aamodt, R CordonCardo, C Fradet, Y Grossman, B Hemstreet, G AF Moore, DH Epstein, L Reeder, J Wheeless, L Waldman, FM Aamodt, R CordonCardo, C Fradet, Y Grossman, B Hemstreet, G TI Interlaboratory variability in fluorescence in situ hybridization analysis SO CYTOMETRY LA English DT Article DE fluorescence in situ hybridization; variability; bladder cancer; chi(2); multinomial distribution ID INSITU HYBRIDIZATION; BLADDER-CANCER; ABERRATIONS AB Reliable interpretation of fluorescence in situ hybridization (FISH) data, especially data that have been generated in more than one laboratory, requires knowledge of the sources of variability inherent in FISH analysis, Possible sources of variation may derive from differences in sample preparation, probes used, intrasample heterogeneity, hybridization protocols, counting criteria within and between scorers, fluorescence microscopes, and filters. This study characterized the relative weight of some of these factors in order to determine the degree to which FISH results are comparable between laboratories, We used a hierarchical partitioned chi(2) analysis to measure sources of variation. We found that replicate counts varied no more than expected based on counting statistics (i.e., multinomial variation), However, with replicate hybridizations done in two separate laboratories, the variability increased significantly, Thus, care must be taken when interpreting FISH data that are derived from more than one institution, Previously agreed upon counting criteria as well as standardized FISH hybridization protocols may decrease this variability. (C) 1996 Wiley-Liss, Inc. C1 NCI,BETHESDA,MD 20892. MEM HOSP,NEW YORK,NY. UNIV LAVAL,QUEBEC CITY,PQ,CANADA. ANDERSON MED CTR,HOUSTON,TX. UNIV OKLAHOMA,MED CTR,OKLAHOMA CITY,OK. UNIV ROCHESTER,ROCHESTER,MI. RP Moore, DH (reprint author), UNIV CALIF SAN FRANCISCO,DEPT EPIDEMIOL,DIV BIOSTAT,MCB 230,BOX 0808,SAN FRANCISCO,CA 94143, USA. OI Reeder, Jay/0000-0002-7125-6893 FU NCI NIH HHS [CA47537] NR 8 TC 11 Z9 11 U1 0 U2 2 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC 605 THIRD AVE, NEW YORK, NY 10158-0012 SN 0196-4763 J9 CYTOMETRY JI Cytometry PD OCT 1 PY 1996 VL 25 IS 2 BP 125 EP 132 DI 10.1002/cyto.990250202 PG 8 WC Biochemical Research Methods; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA VK648 UT WOS:A1996VK64800002 PM 8891442 ER PT J AU Jones, LN Steinert, PM AF Jones, LN Steinert, PM TI Hair keratinization in health and disease SO DERMATOLOGIC CLINICS LA English DT Review ID INTERMEDIATE FILAMENT STRUCTURE; EPIDERMOLYSIS-BULLOSA SIMPLEX; RECESSIVE LAMELLAR ICHTHYOSIS; EPITHELIAL CYTOKERATINS; TRANSGENIC MICE; MAMMALIAN-TISSUES; HUMAN KERATIN-14; GENE-CLUSTER; FATTY-ACIDS; EXPRESSION AB The cells of the epidermis and its derivative, the hair follicle, undergo processes of terminal differentiation that involves the synthesis and assembly of classes of protein and enzymes to form the stratum corneum of the epidermis, and the hair fiber and its cuticle. Using genetic linkage and DNA sequencing methods, we now know that mutations in several genes encoding epidermal keratins or a transglutaminase enzyme cause ichthyosis-related diseases. Similar methods have now suggested that mutations in hair keratin genes underlie some cases of monilethrix, and a deficiency in a cuticle lipid metabolizing enzyme causes maple syrup urine disease. It is to be expected that further application of these methods will elucidate the molecular bases of other genetic hair diseases. C1 NIAMSD,SKIN BIOL LAB,NIH,BETHESDA,MD 20892. NR 105 TC 14 Z9 14 U1 0 U2 1 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0733-8635 J9 DERMATOL CLIN JI Dermatol. Clin. PD OCT PY 1996 VL 14 IS 4 BP 633 EP & DI 10.1016/S0733-8635(05)70390-9 PG 19 WC Dermatology SC Dermatology GA VJ549 UT WOS:A1996VJ54900010 PM 9238322 ER PT J AU Nicolucci, A Carinci, F Graepel, JG Hohman, TC Ferris, F Lachin, JM AF Nicolucci, A Carinci, F Graepel, JG Hohman, TC Ferris, F Lachin, JM TI The efficacy of tolrestat in the treatment of diabetic peripheral neuropathy - A meta-analysis of individual patient data SO DIABETES CARE LA English DT Article ID ALDOSE REDUCTASE INHIBITORS; MULTICENTER TRIAL; FOOT ULCERATION; NATURAL-HISTORY; CLINICAL-TRIALS; NERVE FUNCTION; POLYNEUROPATHY; COMPLICATIONS; MELLITUS; PREVENTION AB OBJECTIVE - The aim of this meta-analysis was to review the existent evidence on the effectiveness of tolrestat in the treatment of diabetic peripheral neuropathy. RESEARCH DESIGN AND METHODS - Individual patient data on 738 subjects from the three randomized clinical trials published on this topic were analyzed using changes in motor nerve conduction velocities (NCVs) as endpoints, Nerves investigated included median, ulnar, tibial, and peroneal, RESULTS - The pooled analysis of NCV taken as a continuous measurement showed a significant treatment effect, the magnitude of this benefit being approximately equal to 1 mis for all the nerves investigated. When looking at the proportion of patients experiencing a loss of NCV of at least 1 or 2 mis in at least two out of the four nerves investigated, it emerged that treatment reduced by >40% the risk of such outcomes after adjusting for patients' characteristics. The odds ratios relative to the placebo group were 1.82 (1.30-2.52) and 1.70 (1.15-2.48) for a decrease of 1 and 2 m/s, that is, placebo-treated patients have an 82 and 70% increased risk for a loss of nerve function of 1 and 2 m/s, respectively. No statistically significant difference in treatment effect emerged after stratification according to baseline motor NCV and glycated hemoglobin levels. CONCLUSIONS - After a treatment duration ranging between 24-52 weeks, patients treated with tolrestat had a reduced risk for developing nerve function loss compared with placebo-treated patients. Future long-term trials are needed to evaluate the impact of the treatment on more clinically meaningful endpoints such as the development of foot complications. C1 WYETH INC,RADNOR,PA. WYETH AYERST RES,PRINCETON,NJ 08543. GEORGE WASHINGTON UNIV,CTR BIOSTAT,BETHESDA,MD. NEI,NIH,BETHESDA,MD 20892. RP Nicolucci, A (reprint author), IST RIC FARMACOL MARIO NEGRI,CONSORZIO MARIO NEGRI SUD,DEPT CLIN PHARMACOL & EPIDEMIOL,VIA NAZL,I-66030 SANTA MARIA IMBAR,CH,ITALY. RI Carinci, Fabrizio/A-1349-2012; OI Lachin, John/0000-0001-9838-2841 NR 30 TC 24 Z9 26 U1 0 U2 0 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 SN 0149-5992 J9 DIABETES CARE JI Diabetes Care PD OCT PY 1996 VL 19 IS 10 BP 1091 EP 1096 DI 10.2337/diacare.19.10.1091 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VK153 UT WOS:A1996VK15300007 PM 8886554 ER PT J AU Gregg, EW Kriska, AM Narayan, KMV Knowler, WC AF Gregg, EW Kriska, AM Narayan, KMV Knowler, WC TI Relationship of locus of control to physical activity among people with and without diabetes SO DIABETES CARE LA English DT Article ID HEALTH LOCUS; PIMA-INDIANS; CARDIAC REHABILITATION; CONTROL BELIEFS; WOMEN; MELLITUS; PERSPECTIVE; BEHAVIOR AB OBJECTIVE - To examine the relationship between locus of control (LOG) (internal and external) and physical activity in Pima Indians and to determine whether this relationship is affected by the presence of diabetes. RESEARCH DESIGN AND METHODS - A population-based sample of 580 Pima Indians was recruited from an ongoing research study. LOC was measured on a 1-40 modified Rotter scale, and past year total physical activity (leisure and work physical activity levels combined) was measured by interviewer-administered questionnaire. RESULTS - Among both men and women without diabetes, individuals with an internal LOC (score 1-16) were significantly (P < 0.01) more active than those with an external (score 17-40) LOC (70 vs. 30 median metabolic equivalent [MET] hours per week for men; 12 vs. 5 median MET hours per week for women). Controlled for age and BMI, an internal LOC was significantly associated with a higher level of physical activity among men (P = 0.04) and women (P = 0.001) without diabetes, but, not among those with diabetes. CONCLUSIONS - Nondiabetic Pima Indians with an internal LOC are more physically active than those with an external LOG. Enhancing perceptions of internal control may influence physical activity and thus have implications for diabetes prevention. C1 UNIV PITTSBURGH,GRAD SCH PUBL HLTH,DEPT EPIDEMIOL,PITTSBURGH,PA. NIDDKD,DIABET & ARTHRIT EPIDEMIOL SECT,PHOENIX,AZ. RI Narayan, K.M. Venkat /J-9819-2012 OI Narayan, K.M. Venkat /0000-0001-8621-5405 FU NIA NIH HHS [32AG00181]; NIDDK NIH HHS [DK43394] NR 33 TC 9 Z9 9 U1 0 U2 0 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 SN 0149-5992 J9 DIABETES CARE JI Diabetes Care PD OCT PY 1996 VL 19 IS 10 BP 1118 EP 1121 DI 10.2337/diacare.19.10.1118 PG 4 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VK153 UT WOS:A1996VK15300012 PM 8886559 ER PT J AU DeSilva, MG Jun, HS Yoon, JW Notkins, AL Lan, MS AF DeSilva, MG Jun, HS Yoon, JW Notkins, AL Lan, MS TI Autoantibodies to IA-2 not detected in NOD mice or BB rats SO DIABETOLOGIA LA English DT Letter ID PROTEIN-TYROSINE-PHOSPHATASE; DIABETES-MELLITUS; ANTIBODIES; ANTIGEN; PRECEDE; ONSET C1 NIDR,ORAL MED LAB,NIH,BETHESDA,MD 20892. NR 14 TC 11 Z9 11 U1 0 U2 2 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 SN 0012-186X J9 DIABETOLOGIA JI Diabetologia PD OCT PY 1996 VL 39 IS 10 BP 1237 EP 1238 DI 10.1007/BF02658513 PG 2 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VK991 UT WOS:A1996VK99100016 PM 8897014 ER PT J AU Lin, YI AF Lin, YI TI Day 11 mouse fetal thymus: Phenotype and search for the point of commitment SO DIFFERENTIATION LA English DT Article ID HEMATOPOIETIC STEM-CELLS; COLONY-FORMING CELLS; MURINE BONE-MARROW; RADIOPROTECTIVE ABILITY; SPLEEN COLONIES; CFU-S; EXPRESSION; SUBSETS; THYMOCYTES; ANTIGEN AB This phenotype study of uncommitted hematopoietic cells (UHC) and early T cell precursors (ETCP) in the thymus provides an understanding of the commitment process from UHC to ETCP. The study of genes that are involved in this process depends on the proper identification of these early cells. Nonetheless, most current phenotype studies of these cells are based on the observations from the late stages of fetal or adult thymocytes. Though conclusions drawn from these studies are insightful, cell maturation occurs so fast during thymus development that some important phenotypic nuances go unnoticed if one only looks at late-stage cells. Furthermore, even though early-stage thymocytes are phenotypically similar to those of late stage, they may have different properties. In order to study those thymus populations at the very beginning of their differentiation and commitment, 11-day mouse fetal thymuses were tested. This shows, using day-12 CFU-S as a measure, that at this stage of development there are UHC present. Thymocytes at this very initial stage of development are isolated for the first time, and their phenotypes as well as their differentiation potentials are analyzed. The result shows that UHC are detected only in the Mac-1(-) C-kit(+) subset, which comprises 2% of the total thymus population. Surprisingly, 93% of the C-kit(+) population is Mac-lf, which generates T cells with a frequency of 1/72. This indicates that the Mac-1 molecule is a differentiation marker rather than a lineage-specific marker. In addition, C-kit(+) thymocytes at this stage do not express any T cell markers, such as Thy1.2, IL-2R alpha, CD2 and Mel-14. These thymocytes are very akin to UHC, since they express Sca-1, Wga, and CD34, and are likewise similar to ETCP, as they express Pgp(high), Mhc-I-high, Hsa(low), and FcR(low). Moreover, they express a high level of adhesion molecules such as Lfa-1, Icam-1 and Lpam-1. As expected, these C-kit(+) all contain the hematopoietic cell marker CD45. Low expression of CD4 (the typical marker associated to the earliest T cell precursor in adult thymuses) is also found in 10% of the C-kit(+) population. While the C-kit(+) population at this stage is more homogeneous than at any other stage of fetal thymus development, there are still markers (B220, CD5, Tsa, Mac-1, CD4, and Sca-1) that can split this population into other subsets. However, the majority of these markers are present in the Mac-1(+) C-kit(+) population, indicating that the C-kit(+) population is essentially made of two populations (Mac-1(+) and Mac-1(-) subsets). Interestingly two major single-positive populations appear to emerge from this C-kit(+) population one day later, namely Thy1.2(+) IL-2R alpha(-) and Thy1.2(-) IL-2R alpha(+). These two major single-positive populations seem to be derived directly from the Mac-1(+) rather than from the Mac-1(-) subset of the C-kit(+) population. Thus, these data suggest that important phenotypes are present during the early differentiation process. These phenotypes have never been shown before. Hopefully, this study will open up a new avenue for the study of very early stage T cell sublineages and their relationship to uncommitted thymic hematopoietic cells. C1 NIAID,CELLULAR & MOL IMMUNOL LAB,BETHESDA,MD 20892. NR 31 TC 2 Z9 2 U1 0 U2 0 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 SN 0301-4681 J9 DIFFERENTIATION JI Differentiation PD OCT PY 1996 VL 61 IS 1 BP 53 EP 65 PG 13 WC Cell Biology; Developmental Biology SC Cell Biology; Developmental Biology GA VQ304 UT WOS:A1996VQ30400006 ER PT J AU Zhan, QM Fan, SJ Smith, ML Bae, I Yu, K Alamo, I OConnor, PM Fornace, AJ AF Zhan, QM Fan, SJ Smith, ML Bae, I Yu, K Alamo, I OConnor, PM Fornace, AJ TI Abrogation of p53 function affects gadd gene responses to DNA base-damaging agents and starvation SO DNA AND CELL BIOLOGY LA English DT Article ID CYCLE CHECKPOINT PATHWAY; TUMOR-SUPPRESSOR P53; CELL-CYCLE; GENOTOXIC STRESS; GROWTH ARREST; ATAXIA-TELANGIECTASIA; BAX GENE; ACTIVATION; EXPRESSION; INDUCTION AB The tumor suppressor p53 is required for induction of its downstream effector genes such as GADD45 and CIP1/WAF1 by ionizing radiation (IR), This response is probably mediated through defined p53 binding sites located in the promoter of CIP1/WAF1 and in the third intron of GADD45. In contrast, the gadd gene stress response to base-damaging agents, such as methylmethane sulfonate (MMS) or UV radiation, or medium depletion (starvation) occurs in all mammalian cells examined to date regardless of p53 status for both GADD45 and also GADD153, which is not IR-responsive in many lines with functional p53, These agents strongly induce the p53 protein and raise the possibility that, although p53 is not required for the typical ''gadd'' response to these agents, p53 may contribute to these non-IR stress responses, This possibility was confirmed by the finding that disruption of p53 function by transfection with dominant-negative vectors expressing HPV E6, mutant p53, or SV40 T Ag reduced the induction of GADD45 and GADD153 as measured by increases in mRNA and protein levels in human lines with wild-type p53, Similarly, induction of these genes by MMS or UV radiation was consistently stronger in the parental mouse embryo fibroblasts compared to cells derived from mice where both p53 alleles had been deleted, Similar qualitative responses were also seen for CIP1/WAF1. In agreement with reduced induction of p53-regulated genes, the G(1) checkpoint activated by MMS or UV radiation was markedly abrogated in p53-wt human MCF-7 breast carcinoma cells by E6 expression, Interestingly, induction of reporter constructs driven by the GADD45 or GADD153 promoters was substantially reduced in human cells transfected with mutant p53 or E6 expression vectors or in cells lacking p53 following treatment with MMS, UV radiation, or starvation, Because neither promoter is inducible by IR, and neither contains a strong p53 binding site, these results indicate that p53 has a synergistic or cooperative role in these non-IR stress responses for both GADD45 and GADD153, and that this role is not mediated through identifiable p53-binding sites. C1 NCI,MOL PHARMACOL LAB,DIV BASIC SCI,NIH,BETHESDA,MD 20892. RI Fornace, Albert/A-7407-2008 OI Fornace, Albert/0000-0001-9695-085X NR 49 TC 90 Z9 94 U1 0 U2 3 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 SN 1044-5498 J9 DNA CELL BIOL JI DNA Cell Biol. PD OCT PY 1996 VL 15 IS 10 BP 805 EP 815 DI 10.1089/dna.1996.15.805 PG 11 WC Biochemistry & Molecular Biology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Cell Biology; Genetics & Heredity GA VQ776 UT WOS:A1996VQ77600002 PM 8892753 ER PT J AU Yoshida, K Cleaveland, ES Nagle, JW French, S Yaswen, L Ohshima, T Brady, RO Pentchev, PG Kulkarni, AB AF Yoshida, K Cleaveland, ES Nagle, JW French, S Yaswen, L Ohshima, T Brady, RO Pentchev, PG Kulkarni, AB TI Molecular cloning of the mouse apolipoprotein D gene and its upregulated expression in Niemann-Pick disease type C mouse model SO DNA AND CELL BIOLOGY LA English DT Article ID RNA TISSUE DISTRIBUTION; BREAST-CANCER CELLS; MESSENGER-RNA; CHOLESTEROL ACYLTRANSFERASE; INSITU HYBRIDIZATION; PERIPHERAL-NERVE; A-I; IDENTIFICATION; DIFFERENTIATION; LOCALIZATION AB Apolipoprotein D (ApoD) is a member of the lipocalin superfamily, The primary structure and diverse expression of ApoD suggest that this protein is a multiligand, multifuuctional glycoprotein, Here we report the structure of the mouse ApoD gene, which is composed of six exons spanning approximately 20 kb in length, All the exon-intron splice junctions follow the consensus GT/AG sequence, The 5'-flanking region of the mouse ApoD gene contains several putative regulatory elements, including FSE-2, GRE, SDR, MRE, IL-6RE, and TATA box, Northern blot analysis revealed that ApoD was highly expressed in the brain and adipose tissue in mouse, Lower levels of expression were observed in the heart, lung, thymus, testis, and salivary glands, In situ hybridization for the brain showed that ApoD mRNA was mainly localized in the subarachnoid space including the pia. In the Niemann-Pick disease type C mouse model, ApoD expression was upregulated in many organs such as brain, adipose tissue, heart, and thymus, presumably due to enhanced ApoD synthesis in perivascular fibroblasts. C1 NIDR, GENE TARGETING RES & CORE FACIL, NIH, BETHESDA, MD 20892 USA. NINCDS, DEV & METAB NEUROL BRANCH, GENET SECT, NIH, BETHESDA, MD 20892 USA. NR 36 TC 23 Z9 25 U1 0 U2 1 PU MARY ANN LIEBERT INC PI NEW ROCHELLE PA 140 HUGUENOT STREET, 3RD FL, NEW ROCHELLE, NY 10801 USA SN 1044-5498 J9 DNA CELL BIOL JI DNA Cell Biol. PD OCT PY 1996 VL 15 IS 10 BP 873 EP 882 DI 10.1089/dna.1996.15.873 PG 10 WC Biochemistry & Molecular Biology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Cell Biology; Genetics & Heredity GA VQ776 UT WOS:A1996VQ77600008 PM 8892759 ER PT J AU Acteo, MD Bowman, E Butelman, E English, JA Harris, L Jacobson, AE Mattson, MV Medzihradsky, F Patrick, G Rowlett, JK Smith, CB Winger, G Woods, JH Woolverton, WL AF Acteo, MD Bowman, E Butelman, E English, JA Harris, L Jacobson, AE Mattson, MV Medzihradsky, F Patrick, G Rowlett, JK Smith, CB Winger, G Woods, JH Woolverton, WL TI Zipeprol: Preclinical assessment of abuse potential SO DRUG AND ALCOHOL DEPENDENCE LA English DT Article DE zipeprol; abuse potential; opioid abuse; piperazine ID RHESUS-MONKEYS; SIGMA-RECEPTORS; BINDING; RAT; CLASSIFICATION; PHENCYCLIDINE; ALFENTANIL; DEPENDENCE; MORPHINE; INFUSION AB Zipeprol was evaluated in a number of in vitro and in vivo assays predictive of stimulant, depressant, or opioid abuse potential. Zipeprol had affinity for mu and kappa opioid binding sites as well as sigma binding sites. However, it failed to exert opioid-like agonist actions in rodents, and did not attenuate withdrawal signs in morphine- or pentobarbital-dependent rats. Zipeprol did not substitute for either amphetamine or pentobarbital in drug discrimination assays in rhesus monkeys. On the other hand, it suppressed morphine withdrawal signs in rhesus monkeys in two assays, and it acted as a quadazocine-sensitive reinforcer in monkeys trained to self-inject alfentanil. Zipeprol also acted as a reinforcer in monkeys trained to self-inject methohexital. In a dose range of 10-18 mg/kg, zipeprol induced convulsions in monkeys. Zipeprol appears to have abuse potential and a novel spectrum of action involving both opioid and non-opioid effects. C1 UNIV MICHIGAN,SCH MED,DEPT PHARMACOL,ANN ARBOR,MI 48109. VIRGINIA COMMONWEALTH UNIV,MED COLL VIRGINIA,DEPT PHARMACOL,RICHMOND,VA 23298. UNIV MICHIGAN,DEPT BIOL CHEM,ANN ARBOR,MI 48109. UNIV MICHIGAN,DEPT PSYCHOL,ANN ARBOR,MI 48109. NIDDK,MED CHEM LAB,NIH,BETHESDA,MD 20892. UNIV MISSISSIPPI,MED CTR,DEPT PSYCHIAT & HUMAN BEHAV,JACKSON,MS 39216. FU NIDA NIH HHS [DA-05951, DA-09139, DA-00254] NR 39 TC 1 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 0376-8716 J9 DRUG ALCOHOL DEPEN JI Drug Alcohol Depend. PD OCT PY 1996 VL 42 IS 2 BP 93 EP 104 DI 10.1016/0376-8716(96)01267-7 PG 12 WC Substance Abuse; Psychiatry SC Substance Abuse; Psychiatry GA VH908 UT WOS:A1996VH90800003 PM 8889408 ER PT J AU Karam, WG Goldstein, JA Lasker, JM Ghanayem, BI AF Karam, WG Goldstein, JA Lasker, JM Ghanayem, BI TI Human CYP2C19 is a major omeprazole 5-hydroxylase, as demonstrated with recombinant cytochrome P450 enzymes SO DRUG METABOLISM AND DISPOSITION LA English DT Article ID S-MEPHENYTOIN 4'-HYDROXYLATION; HUMAN-LIVER; GENETIC-POLYMORPHISM; METABOLISM; HYDROXYLATION; IDENTIFICATION; PHARMACOKINETICS; POPULATION AB Omeprazole (OP) is a potent antiulcer drug that is metabolized by liver cytochrome P450 (P450) enzymes, However, the identities of the P450 isoforms responsible for its metabolism have been controversial, 5-Hydroxyomeprazole (5OH-OP) formation cosegregates with the polymorphism of (S)-mephenytoin 4'-hydroxylation in humans, which is now known to be mediated by CYP2C19, Previous in vitro studies have indicated that liver microsomal 5OH-OP formation correlates with both (S)-mephenytoin 4'-hydroxylase and CYP3A content, Inhibitor and CYP2C antibody studies also suggested that both enzymes may be involved in the 5-hydroxylation of OP, whereas CYP3A appears to be the predominant enzyme involved in OP sulfone (OP-S) formation. The present studies assessed the contribution of various CYP2C and CYP3A4 enzymes to OP metabolism by using recombinant human enzymes, CYP2C19, CYP2C8, CYP2C18, and CYP2C9 formed a single metabolite with an HPLC retention time identical to that of 5OH-OP, The turnover number for CYP2C19 was 13.4 +/- 1.4 nmol/min/nmol of P450, whereas those for CYP2C8, CYP2C18, and CYP2C9 were 2.2 +/- 0.1, 1.5 +/- 0.1, and approximate to 0.5 nmol/min/nmol of P450, respectively, Recombinant human CYP3A4 formed 5OH-OP and OP-S with turnover numbers of 5.7 +/- 1.1 and 7.4 +/- 0.9 nmol/min/nmol of P450, respectively, and formed a minor unidentified metabolite, CYP2C19 had a substantially lower K-M for 5OH-OP formation than did CYP3A4, CYP2C8, or CYP2C18, Antibody to CYP2C proteins inhibited approximate to 70% of OP 5-hydroxylation at low substrate concentrations, comparable to those that may be encountered at therapeutically relevant doses, whereas antibody to CYP3A4 inhibited approximate to 30% of the activity, At high substrate concentrations, the contributions of the two enzymes to OP hydroxylation were roughly comparable (40-50%). In contrast, OP-S formation was completely inhibited by antibody to CYP3A4 proteins. The present study provides the first direct confirmation, using human recombinant P450 enzymes and selective antibody inhibition, that CYP2C19 is a major high affinity OP 5-hydroxylase and CYP3A4 is a low affinity OP-hydroxylating enzyme, The current work also shows, for the first time, that other CYP2C enzymes (CYP2C8, CYP2C9, and CYP2C18) may contribute to OP hydroxylation at high substrate concentrations. In contrast, OP-S was formed principally by CYP3A4. C1 NIEHS,NIH,RES TRIANGLE PK,NC 27709. MT SINAI SCH MED,NEW YORK,NY. RI Goldstein, Joyce/A-6681-2012 NR 29 TC 80 Z9 84 U1 0 U2 4 PU WILLIAMS & WILKINS PI BALTIMORE PA 351 WEST CAMDEN ST, BALTIMORE, MD 21201-2436 SN 0090-9556 J9 DRUG METAB DISPOS JI Drug Metab. Dispos. PD OCT PY 1996 VL 24 IS 10 BP 1081 EP 1087 PG 7 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA VM160 UT WOS:A1996VM16000006 PM 8894508 ER PT J AU Mitsuya, H AF Mitsuya, H TI Ritonavir - A viewpoint SO DRUGS LA English DT Editorial Material RP Mitsuya, H (reprint author), NCI,EXPT RETROVIROL SECT,BETHESDA,MD 20892, USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU ADIS INTERNATIONAL LTD PI AUCKLAND PA 41 CENTORIAN DR, PRIVATE BAG 65901, MAIRANGI BAY, AUCKLAND 10, NEW ZEALAND SN 0012-6667 J9 DRUGS JI Drugs PD OCT PY 1996 VL 52 IS 4 BP 547 EP 548 PG 2 WC Pharmacology & Pharmacy; Toxicology SC Pharmacology & Pharmacy; Toxicology GA VL195 UT WOS:A1996VL19500011 ER PT J AU Wassermann, EM Grafman, J Berry, C Hollnagel, C Wild, K Clark, K Hallett, M AF Wassermann, EM Grafman, J Berry, C Hollnagel, C Wild, K Clark, K Hallett, M TI Use and safety of a new repetitive transcranial magnetic stimulator SO ELECTROMYOGRAPHY AND MOTOR CONTROL-ELECTROENCEPHALOGRAPHY AND CLINICAL NEUROPHYSIOLOGY LA English DT Article DE motor cortex; motor performance; memory; cortical inhibition; brain stimulation ID LOW-FREQUENCY STIMULATION; LONG-TERM POTENTIATION; MOTOR CORTEX; NO EVIDENCE; DEPRESSION; EPILEPSY; HIPPOCAMPUS; MECHANISMS; INDUCTION; PERFORMANCE AB In order to test a new repetitive transcranial magnetic stimulator, the Dantec MagPro, we administered transcranial magnetic stimulation (TMS) at 1 Hz and 125% of motor threshold for an average of 204 s (until the coil temperature reached 40 degrees C) and 20 Hz stimulation at 100% of motor threshold for 2 s every minute for 10 min, on different days to 10 healthy volunteers, We stimulated 6 scalp positions (primary motor area (MI) and sites 5 cm anterior and posterior on each hemisphere) with an 8-shaped coil. We tested immediate and delayed memory, verbal fluency, prolactin levels and EEG at the beginning of the study and after stimulation on each day. No abnormalities were found. Motor evoked potentials evoked with 1 Hz stimulation diminished progressively in amplitude, and 1 Hz stimulation of M1 caused inhibition lasting at least 1 min in 3 of 4 subjects who were tested with 0.1 Hz stimulation before and after the 1 Hz stimulation period. This did not occur with 20 Hz stimulation. Finger tapping frequency was tested at the beginning of the study and after TMS at each scalp site. Finger tapping rare data from 6 additional subjects who were stimulated in an identical fashion with a different stimulator were also analyzed, There was an increase in tapping rate after TMS which was independent of scalp site. This was most pronounced with 1 Hz stimulation at 125% of threshold and reached statistical significance in the hand contralateral to the stimulation. The results of this study indicate that rTMS with the MagPro stimulator is safe at specific combinations of intensity, frequency and train duration. C1 NINCDS,HUMAN MOTOR CONTROL SECT,MED NEUROL BRANCH,NIH,BETHESDA,MD 20892. NINCDS,COGNIT NEUROSCI SECT,MED NEUROL BRANCH,NIH,BETHESDA,MD 20892. OI Grafman, Jordan H./0000-0001-8645-4457 NR 39 TC 185 Z9 192 U1 0 U2 8 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0924-980X J9 ELECTROMYOGR MOTOR C JI Electromyogr. Mot. Control-Electroencephalogr. Clin. Neurophysiol. PD OCT PY 1996 VL 101 IS 5 BP 412 EP 417 PG 6 WC Engineering, Biomedical; Neurosciences SC Engineering; Neurosciences & Neurology GA VT831 UT WOS:A1996VT83100007 PM 8913194 ER PT J AU Janini, GM Muschik, GM Issaq, HJ AF Janini, GM Muschik, GM Issaq, HJ TI Electrokinetic chromatography in suppressed electroosmotic flow environment: Use of a charged cyclodextrin for the separation of enantiomers and geometric isomers SO ELECTROPHORESIS LA English DT Article DE electrokinetic chromatography; charged cyclodextrins; geometric isomers; amino acids; dipeptides; chlorinated phenols; aflatoxins; polyacrylamide-coated columns ID CAPILLARY ZONE ELECTROPHORESIS; HOST-GUEST COMPLEXATION; ETHER-BETA-CYCLODEXTRIN; CHIRAL SEPARATIONS; ANIONIC CYCLODEXTRINS; CHLORINATED PHENOLS; MICELLAR SOLUTIONS; RESOLUTION; PH; ELECTROLYTE AB Electrokinetic chromatography (EKC), with negatively-charged cyclodextrins (NCDs) added to the buffer, was conducted in polyacrylamide-coated columns under suppression of electroosmotic flow. The equations of migration and resolution for neutral solutes in this mode of chromatography, which for brevity we term NCD-EKC, are presented. The chiral sulfated cyclodextrin, beta-CD-SBE (IV), used in this study is anionic over the entire pH range accessible to capillary electrophoresis, and the coated columns are stable and provide reproducible performance in the pH range 2.5-8.8. Optimum separation was obtained in the pH range where the solutes are neutral, The incorporation of an alkyl spacer between the sulfate ion and the rim of the cyclodextrin allows an unhindered approach and inclusion of neutral solutes in the cyclodextrin cavity. Solute migration time is inversely proportional to the concentration of the chiral selector. Separation (relative migration time difference) increases with decreasing chiral selector concentration and approaches a maximum, beyond which further decreases in chiral selector concentration result in broad peaks and loss of resolution. A chiral selector concentration of 1% in a 10 mM phosphate buffer produced excellent separation of amino acids and dipeptide enantiomers. In addition to being chiral selectors, cyclodextrins are also known as shape selectors. NCD-EKC is particularly suited for the separation of positional isomers of hydrophobic solutes. The separation of aflatoxin isomers and chlorophenol congeners is presented. In the separation of chlorophenols the more hydrophobic trichlorophenols eluted first and the least hydrophobic, phenol, eluted last. RP Janini, GM (reprint author), NCI,FREDERICK CANC RES & DEV CTR,SAIC FREDERICK,POB B,FREDERICK,MD 21702, USA. NR 48 TC 55 Z9 56 U1 0 U2 3 PU VCH PUBLISHERS INC PI DEERFIELD BEACH PA 303 NW 12TH AVE, DEERFIELD BEACH, FL 33442-1788 SN 0173-0835 J9 ELECTROPHORESIS JI Electrophoresis PD OCT PY 1996 VL 17 IS 10 BP 1575 EP 1583 DI 10.1002/elps.1150171014 PG 9 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA VV594 UT WOS:A1996VV59400013 PM 8957184 ER PT J AU Lin, KH Zhu, XG Shieh, HY Hsu, HC Chen, ST McPhie, P Cheng, SY AF Lin, KH Zhu, XG Shieh, HY Hsu, HC Chen, ST McPhie, P Cheng, SY TI Identification of naturally occurring dominant negative mutants of thyroid hormone alpha 1 and beta 1 receptors in a human hepatocellular carcinoma cell line SO ENDOCRINOLOGY LA English DT Article ID RETINOIC ACID RECEPTORS; CO-REPRESSOR; MESSENGER-RNA; X RECEPTOR; BINDING; RESISTANCE; TRIIODOTHYRONINE; PROTEIN; DNA; 3,3',5-TRIIODO-L-THYRONINE AB To understand the function of thyroid hormone nuclear receptors (TRs) in human hepatocellular carcinoma cells (HCC), we characterized the hormone binding and transactivational activity of TRs in a HCC cell line, J7. TR alpha 1 (J7-TR alpha 1) and TR beta 1 (J7-TR beta 1) complementary DNAs were cloned from this cell line, and the binding activity to the hormone response elements (TREs) and to the thyroid hormone, 3,3',5-triiodo-L-thyronine (T-3) of the expressed TR proteins were evaluated. J7-TR alpha 1 and J7-TR beta 1 bound to TREs similarly as the TRs isolated from other tissues. However, J7-TR alpha 1 did not bind to T-3, and J7-TR beta 1 bound to T-3 with only about 10% the affinity of the wild-type TR beta 1. Sequencing of the complementary DNAs shows a single Met(259)Ile mutation in J7-TR alpha 1 and Met(334)Val in J7-TR beta 1. Using reporters containing TREs, we found that J7-TR alpha 1 and J7-TR beta 1 had virtually lost their transactivational activity. Moreover, these two mutants inhibited the transactivational activity of the wild-type TRs by a dominant negative effect not only on the transfected TRs, but also on endogenous TRs in other two HCC cell lines, SK-Hep-1 and HepG2. The potency of the dominant negative effect of these two mutants inversely correlated with the expression level of endogenous TRs. The present studies identified two novel naturally occurring TR mutants that have potent dominant negative action. The identification of both the alpha and beta dominant negative mutants in J7 made this cell line a useful model system to further understand the molecular mechanism of the dominant negative action of TR mutants. C1 NCI, DIV BASIC SCI, MOL BIOL LAB, GENE REGULAT SECT, BETHESDA, MD 20892 USA. NIDDK, BIOCHEM PHARMACOL LAB, BETHESDA, MD 20892 USA. RP Lin, KH (reprint author), CHANG GUANG COLL MED & TECHNOL, DEPT BIOCHEM, 259 WEN HWA 1ST RD, TAYUAN, TAIWAN. NR 37 TC 45 Z9 45 U1 0 U2 1 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0013-7227 EI 1945-7170 J9 ENDOCRINOLOGY JI Endocrinology PD OCT PY 1996 VL 137 IS 10 BP 4073 EP 4081 DI 10.1210/en.137.10.4073 PG 9 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VN288 UT WOS:A1996VN28800002 PM 8828459 ER PT J AU Usdin, TB Bonner, TI Harta, G Mezey, E AF Usdin, TB Bonner, TI Harta, G Mezey, E TI Distribution of parathyroid hormone-2 receptor messenger ribonucleic acid in rat SO ENDOCRINOLOGY LA English DT Article ID SMOOTH-MUSCLE CELLS; MEDIAL BASAL HYPOTHALAMUS; PROTEIN PTHRP 1-34; HYPOTENSIVE ACTION; HYPERTENSIVE RATS; GUINEA-PIG; PEPTIDE; BONE; EXPRESSION; KIDNEY AB The PTH2 receptor is a recently identified G protein-coupled receptor activated by PTH. Its amino acid sequence is most similar to the PTH/PTHrP receptor, but unlike the PTH/PTHrP receptor, it is activated by PTH and not by PTH-related peptide. We previously demonstrated using Northern blots that expression of PTH2 receptor messenger RNA was greatest within the brain and occurred at lower levels in pancreas, testis, and placenta. We have now obtained a complementary DNA encoding the rat PTH2 receptor and used it to study the distribution of the PTH2 receptor using in situ hybridization histochemistry. PTH2 receptor messenger RNA is abundantly expressed in arterial and cardiac endothelium and at lower levels in vascular smooth muscle. It is also abundant in the lung, both within bronchi and in the parenchyma, and is present within the exocrine pancreas. It is expressed by sperm in the head of the epididymis. A small number of cells associated with the vascular pole of renal glomeruli express the receptor. These data suggest that the PTH2 receptor may be responsible for PTH effects in a number of physiological systems. C1 NINCDS, LAB CLIN NEUROSCI, NIH, BETHESDA, MD 20892 USA. RP Usdin, TB (reprint author), NIMH, CELL BIOL LAB,NIH,BLDG 36,ROOM 3A17, 36 CONVENT DR, MSC4090, BETHESDA, MD 20892 USA. NR 63 TC 94 Z9 95 U1 0 U2 2 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0013-7227 EI 1945-7170 J9 ENDOCRINOLOGY JI Endocrinology PD OCT PY 1996 VL 137 IS 10 BP 4285 EP 4297 DI 10.1210/en.137.10.4285 PG 13 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VN288 UT WOS:A1996VN28800032 PM 8828488 ER PT J AU Yang, YWH Yanagishita, M Rechler, MM AF Yang, YWH Yanagishita, M Rechler, MM TI Heparin inhibition of insulin-like growth factor-binding protein-3 binding to human fibroblasts and rat glioma cells: Role of heparan sulfate proteoglycans SO ENDOCRINOLOGY LA English DT Article ID BREAST-CANCER CELLS; FACTOR IGF BINDING; FACTOR-I; IGFBP-3; RADIOIMMUNOASSAY; POTENTIATION; PURIFICATION; ASSOCIATION; MECHANISMS; RECEPTORS AB The mitogenic action of insulin-like growth factors (IGFs) on target cells is determined by interaction with signaling IGF-I receptors and modulated by interactions with IGF-binding proteins (IGFBPs). IGFBP-3, an abundant IGFBP that binds IGF-I and IGF-II with high affinity, can form soluble inhibitory complexes with the IGFs that prevent them from binding to IGF-I receptors. Alternatively, IGFBP-3 can bind to the cell surface and possibly potentiate IGF action or act independently of the IGFs. Previous studies showed that heparin inhibited IGFBP-3 binding to the cell surface and increased its accumulation in the medium, suggesting that it might act as a competitive inhibitor of IGFBP-3 binding to structurally similar heparan sulfate proteoglycans on the cell surface. We evaluated this hypothesis by binding I-125-labeled recombinant glycosylated human IGFBP-3 to human fetal skin fibroblasts (GM-10) and to C6 rat glioma cells at 12 C. Heparin inhibited [I-125]IGFBP-3 binding more effectively than chondroitin sulfate and dextran sulfate. Complete digestion of cell surface heparan sulfate and chondroitin sulfate glycosaminoglycans using heparitinase and chondroitinase ABC, however, did not significantly decrease IGFBP-3 binding. Quantitative removal was demonstrated by analysis of parallel cultures of cells whose glycosaminoglycans had been biosynthetically labeled using (Na2SO4)-S-35. These results suggested that IGFBP-3 did not bind to heparan sulfate glycosaminoglycans on the cell surface, and that the inhibition of IGFBP-3 binding by heparin most likely resulted from its direct interaction with the heparin-binding domains of IGFBP-3. When [I-125]IGFBP-3 was incubated with GM-10 fibroblasts or C6 glioma cells at 37 C for 4 h, only 10% of the bound ligand remained associated with the cell surface; approximately 90% of the cell-associated radioactivity was internalized and could he recovered in lysates of acid-washed cells. Incubation with IGF-I or heparin decreased the total cell-associated radioactivity, but did not affect internalization. These results suggest that direct interaction of heparin or IGF-I with IGFBP-3 inhibits its ability to bind to the surface of GM-10 fibroblasts and C6 glioma cells. C1 NIDDK, GROWTH & DEV SECT, MOL & CELLULAR ENDOCRINOL BRANCH, NIH, BETHESDA, MD 20892 USA. NIDR, BONE RES BRANCH, NIH, BETHESDA, MD 20892 USA. NR 41 TC 26 Z9 26 U1 0 U2 1 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0013-7227 EI 1945-7170 J9 ENDOCRINOLOGY JI Endocrinology PD OCT PY 1996 VL 137 IS 10 BP 4363 EP 4371 DI 10.1210/en.137.10.4363 PG 9 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VN288 UT WOS:A1996VN28800041 PM 8828497 ER PT J AU Johns, A Freay, AD Fraser, W Korach, KS Rubanyi, GM AF Johns, A Freay, AD Fraser, W Korach, KS Rubanyi, GM TI Disruption of estrogen receptor gene prevents 17 beta estradiol-induced angiogenesis in transgenic mice SO ENDOCRINOLOGY LA English DT Article ID FUNCTIONALLY AGONADAL WOMEN; POLYSILOXANE VAGINAL RINGS; CYLINDERS; DISEASES; MOUSE AB Estrogen is known to modulate angiogenesis, both under physiological and pathological conditions, and has been demonstrated to augment angiogenesis induced by bFGF in a mouse model. We have modified this mouse model and measured the apparent plasma volume in Matrigel plugs containing basic fibroblast growth factor (bFGF) in wild type and estrogen receptor knockout, ovariectomized mice in the presence and absence of exogenous 17 beta estradiol. The apparent plasma volume was determined by measuring the fluorescence of the excised plug 10 min. after injection of fluoroscein labeled dextran 150. In wild type mice exogenous 17 beta estradiol increased the apparent plasma volume of the Matrigel plug and the uterine weight significantly. In the estrogen receptor knockout mice exogenous 17 beta estradiol caused a small, but significant increase in uterine weight but was without effect on the apparent plasma volume of the Matrigel plug. It is concluded that functional estrogen receptors are essential for the augmentation of bFGF-induced angiogenesis by exogenous 17 beta estradiol in female mice. C1 NIEHS, RECEPTOR BIOL SECT, REPROD & DEV TOXICOL LAB, RES TRIANGLE PK, NC 27709 USA. RP Johns, A (reprint author), BERLEX BIOSCI, CARDIOVASC RES, RICHMOND, CA 94804 USA. OI Korach, Kenneth/0000-0002-7765-418X NR 12 TC 99 Z9 103 U1 0 U2 1 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0013-7227 EI 1945-7170 J9 ENDOCRINOLOGY JI Endocrinology PD OCT PY 1996 VL 137 IS 10 BP 4511 EP 4513 DI 10.1210/en.137.10.4511 PG 3 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VN288 UT WOS:A1996VN28800059 PM 8828515 ER PT J AU Olden, K Guthrie, J AF Olden, K Guthrie, J TI Air toxics regulatory issues facing urban settings SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article; Proceedings Paper CT Symposium on Air Toxics - Biomarkers in Environmental Applications CY APR 27-28, 1995 CL HOUSTON, TX SP Mickey Leland Natl Urban Air Tox Res Ctr, Natl Ctr Environm Hlth, Ctr Dis Control & Prevent, NIEHS, US EPA, Soc Toxicol DE air toxics; biomarkers; regulatory standards; risk assessment AB Biomarker research does not exist in isolation. Its usefulness can only be realized when it is translated into prevention strategies to protect public health. In the context of air toxics, these prevention strategies begin with the development of regulatory standards derived from risk assessment schemes. The Clean Air Act Amendments of 1990 list 189 air toxics, including many volatile organics, metals, and pesticides. The National Institute of Environmental Health Sciences (NIEHS), through its affiliation with the National Toxicology Program, has generated toxicity and carcinogenicity data on more than 100 of these air toxics. The NIEHS extramural and intramural research portfolios support a variety of projects that develop and validate biomarkers for use in environmental health science and risk assessment. Biomarkers have a tremendous potential in the areas of regulating air toxics and protecting public health. Risk assessors need data provided by biomarkers of exposure, biomarkers of dose/pharmacokinetics, biomarkers of susceptibility or individual variability, and biomarkers of effects. The greatest benefit would be realized if biomarkers could be employed in four areas of primary and secondary prevention. The first is the use of biomarkers to enhance extrapolation of animal data to human exposure situations in establishing risk standards. The second is the use of biomarkers that assess noncancer. as well as cancer, end points. Important health end points include pulmonary dysfunction, immunotoxicity, and neurotoxicity. Third, biomarkers that serve as early warning signs to detect intermediate effects would enhance our ability to design timely and cost-effective intervention strategies. Finally, biomarkers used to evaluate the effectiveness of intervention strategies, both in clinical and regulatory settings, would enable us to ensure that programs designed to protect public health do, in fact, achieve the desired outcome. C1 NIEHS,RES TRIANGLE PK,NC 27709. NR 7 TC 1 Z9 1 U1 1 U2 2 PU NATL INST ENVIRON HEALTH SCI PI RES TRIANGLE PK PA PO BOX 12233, RES TRIANGLE PK, NC 27709 SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD OCT PY 1996 VL 104 SU 5 BP 857 EP 860 DI 10.2307/3433002 PG 4 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA VU005 UT WOS:A1996VU00500002 PM 8933026 ER PT J AU Poirier, MC Weston, A AF Poirier, MC Weston, A TI Human DNA adduct measurements: State of the art SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article DE polycyclic aromatic hydrocarbons; occupational exposure; ambient benzo[a]pyrene; enzyme-linked immunosorbent assay; P-32-postlabeling; fluorescence spectroscopy; gas chromatography mass spectrometry; biomarkers ID AROMATIC HYDROCARBON-DNA; P-32 POSTLABELING ASSAY; DIOL-EPOXIDE-DNA; WHITE BLOOD-CELLS; COKE-OVEN WORKERS; PERFORMANCE LIQUID-CHROMATOGRAPHY; CANCER-PATIENTS; FOUNDRY WORKERS; MOLECULAR DOSIMETRY; HEMOGLOBIN ADDUCTS AB Human DNA adduct formation (covalent modification of DNA with chemical carcinogens) is a promising biomarker for elucidating the molecular epidemiology of cancer. Classes of compounds for which human DNA adducts have been observed include polycyclic aromatic hydrocarbons (PAHs), nitrosamines, mycotoxins, aromatic amines, heterocyclic amines, ultraviolet light. and alkylating cancer chemotherapeutic agents. Most human DNA adduct exposure monitoring has been performed with either P-32-postlabeling or immunoassays. neither of which is able to chemically characterize specific DNA adducts. Recently developed combinations of methods with chemical and physical end points have allowed identification of specific adducts in human tissues. Studies are presented that demonstrate that high ambient levels of benzo[a]pyrene are associated with high levels of DNA adducts in human blood cell DNA and that the same DNA adduct levels drop when the ambient PAH levels decrease significantly. DNA adduct dosimetry, which has been achieved with some dietary carcinogens and cancer chemotherapeutic agents, is described, as well as studies correlating DNA adducts with other biomarkers. It is likely that some toxic, noncarcinogenic compounds may have genotoxic effects, including oxidative damage, and that adverse health outcomes other than cancer may be correlated with DNA adduct formation. The studies presented here may serve as useful prototypes for exploration of other toxicological end points. C1 MT SINAI SCH MED, DEPT COMMUNITY MED, NEW YORK, NY USA. RP Poirier, MC (reprint author), NCI, CELLULAR CARCINOGENESIS & TUMOR PROMOT LAB, NIH, BLDG 37, ROOM 3B25, BETHESDA, MD 20892 USA. NR 156 TC 59 Z9 60 U1 1 U2 4 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 OCT PY 1996 VL 104 SU 5 BP 883 EP 893 DI 10.2307/3433006 PG 11 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA VU005 UT WOS:A1996VU00500006 PM 8933030 ER PT J AU Talaska, G Underwood, P Maier, A Lewtas, J Rothman, N Jaeger, M AF Talaska, G Underwood, P Maier, A Lewtas, J Rothman, N Jaeger, M TI Polycyclic aromatic hydrocarbons (PAHs), nitro-PAHs and related environmental compounds: Biological markers of exposure and effects SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article DE polycyclic aromatic hydrocarbons; lung cancer; biomarkers; DNA adducts; lymphocytes; urothelial cells ID EXFOLIATED UROTHELIAL CELLS; CARCINOGEN-DNA ADDUCTS; WHITE BLOOD-CELLS; ORGANIC-MATTER; 1-NITROPYRENE; METABOLISM; SMOKING; LUNG; PARTICULATE; MUTAGENS AB Lung cancer caused by polycyclic aromatic hydrocarbons (PAHs), nitro-PAHs and related environmental agents is a major problem in industrialized nations. The high case-fatality rate of the disease, even with the best supportive treatment, underscores the importance of primary lung cancer prevention. Development of biomarkers of exposure and effects to PAHs and related compounds is now underway and includes measurement of urinary metabolites of specific PAHs as well as detection of protein and DNA adducts as indicators of effective dose. Validation of these markers in terms of total environmental dose requires that concurrent measures of air levels and potential dermal exposure be made. In addition, the interrelationships between PAH biomarkers must be determined, particularly when levels of the marker in surrogate molecules (e.g., protein) or markers from surrogate tissues (e.g., lymphocyte DNA) are used to assess the risk to the target organ, the lung. Two approaches to biomarker studies will be reviewed in this article: the progress made using blood lymphocytes as surrogates for lung tissues and the progress made developing noninvasive markers of carcinogen-DNA adduct levels in lung-derived cells available in bronchial-alveolar lavage and in sputum. Data are presented from studies in which exfoliated urothelial cells were used as a surrogate tissue to assess exposure to human urinary bladder carcinogens in occupational groups. C1 US EPA, RES TRIANGLE PK, NC 27711 USA. NCI, BETHESDA, MD 20892 USA. RP Talaska, G (reprint author), UNIV CINCINNATI, SCH MED, DEPT ENVIRONM HLTH, CINCINNATI, OH 45267 USA. FU NIEHS NIH HHS [1P30 ES06096] NR 38 TC 20 Z9 20 U1 5 U2 21 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 OCT PY 1996 VL 104 SU 5 BP 901 EP 906 DI 10.2307/3433008 PG 6 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA VU005 UT WOS:A1996VU00500008 PM 8933032 ER PT J AU Jameson, CW AF Jameson, CW TI Introduction to the Conference on Beryllium-Related Diseases SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Editorial Material RP Jameson, CW (reprint author), NIEHS,POB 12233,RES TRIANGLE PK,NC 27709, USA. NR 8 TC 6 Z9 6 U1 0 U2 0 PU NATL INST ENVIRON HEALTH SCI PI RES TRIANGLE PK PA PO BOX 12233, RES TRIANGLE PK, NC 27709 SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD OCT PY 1996 VL 104 SU 5 BP 935 EP 936 PG 2 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA VU005 UT WOS:A1996VU00500013 PM 8933037 ER PT J AU Bristol, DW Wachsman, JT Greenwell, A AF Bristol, DW Wachsman, JT Greenwell, A TI The NIEHS Predictive-Toxicology Evaluation Project SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Editorial Material DE predictive toxicology; carcinogenesis; decision support; hazard identification; activity classification; risk assessment; pattern recognition; human heuristic; expert system; machine learning; artificial intelligence ID CHEMICAL CARCINOGENESIS; RODENT CARCINOGENICITY; PROGRAM AB The Predictive-Toxicology Evaluation (PTE) project conducts collaborative experiments that subject the performance of predictive-toxicology (PT) methods to rigorous, objective evaluation in a uniquely informative manner. Sponsored by the National Institute of Environmental Health Sciences, it takes advantage of the ongoing testing conducted by the U.S. National Toxicology Program (NTP) to estimate the true error of models that have been applied to make prospective predictions on previously untested, noncongeneric-chemical substances. The PTE project first identifies a group of standardized NTP chemical bioassays either scheduled to be conducted or are ongoing, but not yet complete. The project then announces and advertises the evaluation experiment, disseminates information about the chemical bioassays, and encourages researchers from a wide variety of disciplines to publish their predictions in peer-reviewed journals, using whatever approaches and methods they feel are best. A collection of such papers is published in this Environmental Health Perspectives Supplement, providing readers the opportunity to compare and contrast PT approaches and models, within the context of their prospective application to an actual-use situation. This introduction to this collection of papers on predictive toxicology summarizes the predictions made and the final results obtained for the 44 chemical carcinogenesis bioassays of the first PTE experiment (PTE-1) and presents information that identifies the 30 chemical carcinogenesis bioassays of PTE-2, along with a table of prediction sets that have been published to date. It also provides background about the origin and goals of the PTE project. outlines the special challenge associated with estimating the true error of models that aspire to predict open-system behavior, and summarizes what has been learned to date. C1 NIEHS, ENVIRONM TOXICOL PROGRAM, RES TRIANGLE PK, NC 27709 USA. RP Bristol, DW (reprint author), NIEHS, CANC BIOL GRP, LAB ENVIRONM CARCINOGENESIS & MUTAGENESIS, DIV INTRAMURAL RES,POB 12233, B3-09, RES TRIANGLE PK, NC 27709 USA. NR 28 TC 59 Z9 59 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 EI 1552-9924 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD OCT PY 1996 VL 104 SU 5 BP 1001 EP 1010 PG 10 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA VU005 UT WOS:A1996VU00500024 PM 8933048 ER PT J AU Olden, K AF Olden, K TI Thirty years of environmental health research - And growing SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Editorial Material C1 NIH,NATL TOXICOL PROGRAM,BETHESDA,MD 20892. RP Olden, K (reprint author), NIEHS,POB 12233,RES TRIANGLE PK,NC 27709, USA. NR 0 TC 0 Z9 0 U1 0 U2 2 PU NATL INST ENVIRON HEALTH SCI PI RES TRIANGLE PK PA PO BOX 12233, RES TRIANGLE PK, NC 27709 SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD OCT PY 1996 VL 104 IS 10 BP 1010 EP 1011 DI 10.2307/3433099 PG 2 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA VR978 UT WOS:A1996VR97800001 PM 8930536 ER PT J AU Goyer, RA AF Goyer, RA TI Results of lead research: Prenatal exposure and neurological consequences SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article DE blood lead levels; fetal brain; fetal susceptibility; lead; mechanisms of central nervous system effects; neurological effects; pregnancy; prenatal exposure; risk factors ID PORT-PIRIE COHORT; BLOOD LEAD; NATIONAL-HEALTH; RAT; LEVEL; AGE; NEUROTOXICITY; TOXICITY; SYSTEM AB The history of advances in the understanding of the toxic effects of lead over the past 20 years is an outstanding example of how knowledge learned from research can impact public health. Measures that have had the greatest impact on reducing exposure to lead are reduction of lead from gasoline, elimination of lead solder from canned food, removal of lead from paint, and abatement of housing containing lead-based paint. Nevertheless, continuing factors that enhance risk to lead exposure, particularly during fetal life, are low socioeconomic status, old housing with lead-containing paint, and less than ideal nutrition, particularly low dietary intake of calcium, iron, and zinc. Prenatal exposure may result from endogenous sources such as lead in the maternal skeletal system or maternal exposures from diet and the environment. Experimental studies have shown that the developing nervous system is particularly sensitive to the toxic effects of lead and that a large number of the effects in the nervous system are due to interference of lead with biochemical functions dependent on calcium ions and impairment of neuronal connections dependent on dendritic pruning. There is need for more study to determine whether these effects are a continuum of prenatal lead exposure or whether prenatal exposure to lead produces unique effects. RP Goyer, RA (reprint author), NIEHS,POB 12233,RES TRIANGLE PK,NC 27709, USA. NR 53 TC 110 Z9 118 U1 2 U2 14 PU NATL INST ENVIRON HEALTH SCI PI RES TRIANGLE PK PA PO BOX 12233, RES TRIANGLE PK, NC 27709 SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD OCT PY 1996 VL 104 IS 10 BP 1050 EP 1054 DI 10.2307/3433116 PG 5 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA VR978 UT WOS:A1996VR97800017 PM 8930545 ER PT J AU Reif, JS Hatch, MC Bracken, M Holmes, LB Schwetz, BA Singer, PC AF Reif, JS Hatch, MC Bracken, M Holmes, LB Schwetz, BA Singer, PC TI Reproductive and developmental effects of disinfection by-products in drinking water SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article DE birth defects; chlorination; drinking water; epidemiology; low birth weight; prematurity; reproduction; teratogens; trihalomethanes ID NEURAL-TUBE DEFECTS; PREGNANCY OUTCOMES; DELAYED CONCEPTION; TAP WATER; CONSUMPTION; ASSOCIATION; EXPOSURE; INFANTS AB Recent epidemiologic studies have reported associations between the consumption of chlorinated drinking water and reproductive and development effects. Here we review the available epidemiologic data, assess the hazard potential posed by exposure to disinfection by-products, identify critical data gaps, and offer recommendations for further research. The epidemiologic evidence supporting associations between exposure to water disinfection by-products (DBPs) and adverse pregnancy outcomes is sparse, and positive findings should be interpreted cautiously. The methods used during the early stages of research in this area have been diverse. Variability in exposure assessment and endpoints makes it difficult to synthesize or combine the available data. Exposure misclassification and unmeasured confounding may have lead to bias in risk estimation. Future studies of reproductive outcome and exposure to chlorinated water should use improved methods for exposure assessment to 1) assure selection of appropriate exposure markets, 2) assess seasonal and annual fluctuations in DBPs, 3) assess variability within the distribution system, and 4) assess exposure through multiple routes such as bathing and showering, as well as consumption. Population-based studies should be conducted to evaluate male and female fertility, conception delay, growth retardation, and specific birth defects. The reproductive and development effects of exposure to DBPs could be efficiently explored in ongoing investigations by incorporating valid exposure markers and relevant questionnaire information. Future studies should make use of naturally occurring variability in the concentrations of DBPs and may incorporate biomarkers of exposure and effect in their design. Epidemiologic investigations should be conducted in parallel with laboratory-based and animal studies in a coordinated, multidisciplinary approach. C1 COLORADO STATE UNIV,DEPT ENVIRONM HLTH,FT COLLINS,CO 80523. COLUMBIA UNIV,DIV EPIDEMIOL,NEW YORK,NY 10032. YALE UNIV,DEPT EPIDEMIOL & PUBL HLTH,NEW HAVEN,CT 06510. MASSACHUSETTS GEN HOSP,GENET & TERATOL UNIT,BOSTON,MA 02114. NIEHS,ENVIRONM TOXCIOL PROGRAM,RES TRIANGLE PK,NC 27709. UNIV N CAROLINA,DEPT ENVIRONM SCI & ENGN,CHAPEL HILL,NC 27599. NR 26 TC 64 Z9 64 U1 0 U2 6 PU NATL INST ENVIRON HEALTH SCI PI RES TRIANGLE PK PA PO BOX 12233, RES TRIANGLE PK, NC 27709 SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD OCT PY 1996 VL 104 IS 10 BP 1056 EP 1061 DI 10.2307/3433117 PG 6 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA VR978 UT WOS:A1996VR97800018 PM 8930546 ER PT J AU Albert, RE French, JE Maronpot, R Spalding, J Tennant, R AF Albert, RE French, JE Maronpot, R Spalding, J Tennant, R TI Mechanism of skin tumorigenesis by contact sensitizers: The effect of the corticosteroid fluocinolone acetonide on inflammation and tumor induction by 2,4 dinitro-1-fluorobenzene in the skin of the TG.AC (v-Ha-ras) mouse SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article DE contact sensitization; corticosteroids; dinitrofluorobenzene; fluocinolone acetonide; TG.AC mouse; tumorigenesis; tumor promotion ID PROMOTING ACTIVITY; CARCINOGENICITY; MICE; 2,4-DINITROFLUOROBENZENE; HYPERSENSITIVITY; MUTAGENICITY; CANCER; MODEL AB The effect of the corticosteroid fluocinolone acetonide (FA) on skin tumor induction and inflammation by the contact sensitizer dinitrofluorobenzene (DNFB) was examined. This study broadly relates to the question of whether contact sensitizers, as electrophilic chemicals that produce protein adduction, may constitute an environmental cancer hazard. The specific aim of this study was to evaluate the extent to which the immunogenic inflammatory response to DNFB, in contrast to DNFB cytotoxicity, might be responsible for tumor induction. Experiments were conducted on a transgenic (TG.AC) mouse, incorporating a mutated ras oncogene (v-Ha-ras) that responds rapidly and profusely with skin papillomas to tumor promoters as if it were genetically initiated. Various doses and patterns of DNFB and FA were applied to the skin in a 2-week period; DNFB was given four times and FA was given either with the DNFB or daily. The tumor response to DNFB was completed by 8 weeks from the first dose and was consistent with a dose-squared relationship. FA was not tumorigenic alone; when given with DNFB, it caused only a small reduction in inflammation and tumor yield. When given daily, FA increased ulcerative skin damage, inflammation and tumor yield of tumors. The results suggest that tumorigenesis by DNFB, in the high-dose short-term regimen used here, is mainly due to its cytotoxicity and not contact sensitization. C1 NIEHS,RES TRIANGLE PK,NC 27709. RP Albert, RE (reprint author), UNIV CINCINNATI,MED CTR,DEPT ENVIRONM HLTH,POB 670056,CINCINNATI,OH 45267, USA. NR 44 TC 9 Z9 9 U1 0 U2 1 PU NATL INST ENVIRON HEALTH SCI PI RES TRIANGLE PK PA PO BOX 12233, RES TRIANGLE PK, NC 27709 SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD OCT PY 1996 VL 104 IS 10 BP 1062 EP 1068 DI 10.2307/3433118 PG 7 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA VR978 UT WOS:A1996VR97800019 PM 8930547 ER PT J AU Tennant, RW Spalding, J AF Tennant, RW Spalding, J TI Predictions for the outcome of rodent carcinogenicity bioassays: Identification of trans-species carcinogens and noncarcinogens SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article DE predicting carcinogens; carcinogenicity bioassays; genotoxicity; predictive toxicology; carcinogens ID 44 CHEMICALS; ALPHA-2U-GLOBULIN; NEPHROPATHY; HAZARD AB Thirty chemicals or substances currently undergoing long-term carcinogenicity bioassays in rodents have been used in a project to further evaluate methods and information that may have the capability of predicting potential carcinogens. In our predictions the principal information used includes structural alerts and in vitro test results for Salmonella mutagenicity, relative subchronic toxicity, and the sites and types of pathology found in subchronic (90-day) studies. This group of chemicals differs significantly from those used previously to evaluate predictive methods in that 23 of 30 are defined as nonmutagenic by conventional criteria. The goal of this predictive effort is to identify categorically the chemicals that have the capacity to induce cancers in both rats and mice (trans-species carcinogens) and those that are not carcinogenic in either rats or mice. Chemicals that show properties that may be associated with tumor induction in either species, i.e., species-specific cancers, are categorized as being of ''uncertain predictability.'' This category includes chemicals believed to have limited carcinogenic potential that is manifested principally as a consequence of the genetic background of the test strain of inbred rodent. RP Tennant, RW (reprint author), NIEHS, LAB ENVIRONM CARCINOGENESIS & MUTAGENESIS, POB 12233, RES TRIANGLE PK, NC 27709 USA. NR 15 TC 17 Z9 17 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 EI 1552-9924 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD OCT PY 1996 VL 104 SU 5 BP 1095 EP 1100 DI 10.2307/3433035 PG 6 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA VU005 UT WOS:A1996VU00500035 PM 8933059 ER PT J AU Huff, J Weisburger, E Fung, VA AF Huff, J Weisburger, E Fung, VA TI Multicomponent criteria for predicting carcinogenicity: Dataset of 30 NTP chemicals SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Article DE carcinogenicity; prediction criteria; NTP chemicals ID DROPLET ALPHA-2U-GLOBULIN NEPHROPATHY; RENAL CARCINOGENESIS; CELL-PROLIFERATION; ALTERNATIVE HYPOTHESIS; RISK ASSESSMENT; CANCER; PERSPECTIVE; MECHANISMS; TOXICITY; BENZENE AB This article is in response to the challenge issued to the scientific community by the National Toxicology Program to predict the carcinogenicity potential of 30 chemicals previously selected for long-term carcinogenicity testing. Utilizing the available toxicologic, genetic. and structural information on 30 chemicals previously selected for long-term carcinogenicity testing, we predict that 16 chemicals (53%) would induce some indication of carcinogenic activity in rodents; we further predict that 10 chemicals (33%) would be associated with weak or equivocal carcinogenic responses, and another 4 (13%) would give no indication of carcinogenicity. Our level of certainty is indicated for many of these predictions. Nonetheless, we believe that most instances of guessing whether a chemical would eventually induce cancer in experimental animals and hence represent a carcinogenic hazard to humans are fraught with considerable uncertainty: uncertainty that can only be relieved by long-term testing for carcinogenicity in animals or by conducting an epidemiologic investigation of exposed individuals or groups. We further believe that the day may come when our predictive acumen will be upgraded to such an extent that we might eventually obviate cancer testing. Until then, and in the best interests of public health, however, we urge long term testing of chemicals in animals be continued, at increased pace. C1 NCI, NIH, BETHESDA, MD 20892 USA. RP Huff, J (reprint author), NIEHS, NIH, POB 12233, RES TRIANGLE PK, NC 27709 USA. NR 56 TC 13 Z9 13 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 EI 1552-9924 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD OCT PY 1996 VL 104 SU 5 BP 1105 EP 1112 DI 10.2307/3433037 PG 8 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA VU005 UT WOS:A1996VU00500037 PM 8933061 ER PT J AU Boinski, S Campbell, AF AF Boinski, S Campbell, AF TI The huh vocalization of white-faced capuchins: A spacing call disguised as a food call? SO ETHOLOGY LA English DT Article ID INDIVIDUAL SPATIAL CHOICE; GOLDEN LION TAMARINS; CEBUS-CAPUCINUS; FORAGING GROUPS; SQUIRREL-MONKEYS; TROOP MOVEMENT; VOCAL BEHAVIOR; COMMUNICATION; FIELD; AVAILABILITY AB White-faced capuchins, Cebus capucinus, predictably emit huh vocalizations It high rates within dense fruit patches. We sought to determine why white faced capuchins at the La Selva Biological Station, Costa Rica produce these food-associated calls. Here we analyze the contests in which this intra-group vocalization was emitted, including the spatial responses elicited from other troop members. A cumulative 26.6 h of continuous focal samples and 3314 spectrograms (including 1643 huhs) were analyzed from a study troop with 16 focal subjects. The mean individual rate of huhs was greater (1) during foraging versus nonforaging activities; (2) during fruit foraging compared to both visual searching for foraging sites and foraging for arthropod prey; and (3) when the nearest neighbor was within a 10 m radius of the focal animal compared to when the nearest neighbor was at greater distances. A huh also predicted a significant increase in nearest-neighbour distance; on average, mean nearest-neighbor distance increased 3 m within 2 min following a huh vocalization. Null models of change in mean nearest-neighbor distance over time were generated from the original data set by treating predetermined time points (140 s intervals) in the focal recordings as if those points marked instances at which huhs were produced by the focal subject. No significant alterations in nearest-neighbor distance were detected within time lags up to 100 s in these null models, supporting the conclusion that huhs are causally linked with subsequent increases in nearest-neighbor distances. Huhs were most evident when capuchins were within dense fruit patches, hut these calls were produced across all foraging contests. Our results suggest that huhs may not be food calls in the usual sense (i.e. informing others of the location of food sources to be shared), but may be more appropriately described as spacing calls. Huhs probably act to increase foraging efficiency by reducing overlap in foraging areas with other troop members. C1 UNIV FLORIDA,DIV COMPARAT MED,GAINESVILLE,FL 32611. NATL INST HLTH,COMPARAT ETHOL LAB,BETHESDA,MD. WASHINGTON UNIV,DEPT ANTHROPOL,ST LOUIS,MO 63130. RP Boinski, S (reprint author), UNIV FLORIDA,DEPT ANTHROPOL,1350 TURLINGTON,GAINESVILLE,FL 32611, USA. NR 45 TC 43 Z9 43 U1 0 U2 6 PU BLACKWELL WISSENSCHAFTS-VERLAG GMBH PI BERLIN PA KURFURSTENDAMM 57, D-10707 BERLIN, GERMANY SN 0179-1613 J9 ETHOLOGY JI Ethology PD OCT PY 1996 VL 102 IS 10 BP 826 EP 840 PG 15 WC Psychology, Biological; Behavioral Sciences; Zoology SC Psychology; Behavioral Sciences; Zoology GA VP780 UT WOS:A1996VP78000004 ER PT J AU Lubkowski, J Palm, GJ Gilliland, GL Derst, C Rohm, KH Wlodawer, A AF Lubkowski, J Palm, GJ Gilliland, GL Derst, C Rohm, KH Wlodawer, A TI Crystal structure and amino acid sequence of Wolinella succinogenes L-asparaginase SO EUROPEAN JOURNAL OF BIOCHEMISTRY LA English DT Article DE L-asparaginase; cloning; sequence; crystal structure; Wolinella succinogenes ID ACUTE LYMPHOBLASTIC-LEUKEMIA; ESCHERICHIA-COLI; GLUTAMINASE-ASPARAGINASE; INTENSIVE ASPARAGINASE; VIBRIO-SUCCINOGENES; PURIFICATION; MUTAGENESIS; INDUCTION; THERAPY AB The amino acid sequence and tertiary structure of Wolinella succinogenes L-asparaginase were determined, and were compared with the structures of other type-II bacterial L-asparaginases. Each chain of this homotetrameric enzyme consists of 330 residues. The amino acid sequence is 40-50% identical to the sequences of related proteins from other bacterial sources, and all residues previously shown to be crucial for the catalytic action of these enzymes are identical. Differences between the amino acid sequence of W. succinogenes L-asparaginase and that of related enzymes are discussed in terms of the possible influence on the substrate specificity. The overall fold of the protein subunit is almost identical to that observed for other L-asparaginases. Two fragments in each subunit, a very highly flexible loop (approximate to 20 amino acids) that forms part of the active site, and the N-terminus (two amino acids), are not defined in the structure. The orientation of Thr14, a residue probably involved in the catalytic activity, indicates the absence of ligand in the active-site pocket. The rigid part of the active site, which includes the asparaginase triad Thr93-Lys166-Asp94, is structurally very highly conserved with equivalent regions found in other type-II bacterial L-asparaginases. C1 UNIV MARYLAND,MARYLAND BIOTECHNOL INST,CTR ADV RES BIOTECHNOL,ROCKVILLE,MD 20850. NIST,ROCKVILLE,MD 20850. UNIV MARBURG,INST PHYSIOL CHEM,D-3550 MARBURG,GERMANY. RP Lubkowski, J (reprint author), NCI,FREDERICK CANC RES & DEV CTR,ABL BASIC RES PROGRAM,MACROMOL STRUCT LAB,FREDERICK,MD 21702, USA. NR 39 TC 70 Z9 78 U1 0 U2 4 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 SN 0014-2956 J9 EUR J BIOCHEM JI Eur. J. Biochem. PD OCT PY 1996 VL 241 IS 1 BP 201 EP 207 DI 10.1111/j.1432-1033.1996.0201t.x PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VL748 UT WOS:A1996VL74800028 PM 8898907 ER PT J AU Ambrosone, CB Kato, S Bowman, ED Harrington, AM Blomeke, B Freudenheim, JL Graham, S Marshall, JR Vena, JE Brasure, JR Shields, PG AF Ambrosone, CB Kato, S Bowman, ED Harrington, AM Blomeke, B Freudenheim, JL Graham, S Marshall, JR Vena, JE Brasure, JR Shields, PG TI Molecular epidemiology of lung and breast cancer SO EUROPEAN JOURNAL OF CANCER PREVENTION LA English DT Article; Proceedings Paper CT International Symposium on Causes of Human Cancer CY JUN 03-05, 1996 CL UDINE, ITALY ID P-32 POSTLABELING ASSAY; HYDROCARBON-DNA ADDUCTS; GENETIC POLYMORPHISMS C1 SUNY BUFFALO,DEPT SOCIAL & PREVENT MED,BUFFALO,NY 14260. RP Ambrosone, CB (reprint author), NCI,HUMAN CARCINOGENESIS LAB,BETHESDA,MD 20892, USA. NR 6 TC 3 Z9 3 U1 1 U2 2 PU RAPID SCIENCE PUBLISHERS PI LONDON PA 2-6 BOUNDARY ROW, LONDON, ENGLAND SE1 8NH SN 0959-8278 J9 EUR J CANCER PREV JI Eur. J. Cancer Prev. PD OCT PY 1996 VL 5 IS 5 BP 391 EP 392 PG 2 WC Oncology SC Oncology GA VX947 UT WOS:A1996VX94700023 PM 8972270 ER PT J AU Stojilkovic, SS AF Stojilkovic, SS TI Endothelin receptors and gonadal function: An invited commentary SO EUROPEAN JOURNAL OF ENDOCRINOLOGY LA English DT Editorial Material ID RAT LEYDIG-CELLS; GRANULOSA-CELLS; PROLACTIN SECRETION; INHIBITOR; OVARY RP Stojilkovic, SS (reprint author), NICHHD,ENDOCRINOL & REPROD RES BRANCH,UCS,NIH,BLDG 49,ROOM 6A-36,49 CONVENT DR,MSC 4510,BETHESDA,MD 20892, USA. NR 18 TC 2 Z9 2 U1 0 U2 0 PU SCANDINAVIAN UNIVERSITY PRESS PI OSLO PA PO BOX 2959 TOYEN, JOURNAL DIVISION CUSTOMER SERVICE, N-0608 OSLO, NORWAY SN 0804-4643 J9 EUR J ENDOCRINOL JI Eur. J. Endocrinol. PD OCT PY 1996 VL 135 IS 4 BP 391 EP 393 PG 3 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VR288 UT WOS:A1996VR28800002 PM 8921818 ER PT J AU McDermott, MF SchmidtWolf, G Sinha, AA Koo, M Porter, MA Briant, L CambonThomsen, A Maclaren, NK Fiske, D Bertera, S Trucco, M Amos, CI McDevitt, HO Kastner, DL AF McDermott, MF SchmidtWolf, G Sinha, AA Koo, M Porter, MA Briant, L CambonThomsen, A Maclaren, NK Fiske, D Bertera, S Trucco, M Amos, CI McDevitt, HO Kastner, DL TI No linkage or association of telomeric and centromeric T-cell receptor beta-chain markers with susceptibility to type 1 insulin-dependent diabetes in HLA-DR4 multiplex families SO EUROPEAN JOURNAL OF IMMUNOGENETICS LA English DT Article ID GLUTAMIC-ACID DECARBOXYLASE; MELLITUS IDDM; AUTOIMMUNE-DISEASE; HLA-DQ; GENES; POLYMORPHISMS; COMPLEX; LOCUS; ALPHA; MICE AB The T-cell receptor beta locus (TCRB) on chromosome 7q35 was studied as a candidate region for genetic susceptibility to type 1 insulin-dependent diabetes (IDDM). A highly polymorphic microsatellite marker mapping to the TCRBV6.7 gene and a TCRB C-region RFLP were used to genotype the members of a total of 21 multiplex IDDM families from two different geographical areas. There was no evidence to support linkage to either of these markers with IDDM, and conventional two-point analysis excluded linkage to the telomeric end of the TCRB complex, in the region of the highly informative TCRBV6.7 marker. There was significant linkage of IDDM to the class II HLA-D locus with significant lod scores >3.0 obtained for the HLA-DRB1 and HLA-DQB1 genes. Affected sib-pair (ASP) and transmission disequilibrium (TDT) association tests confirmed these findings. C1 NIAMSD,ARTHRIT & RHEUMATISM BRANCH,BETHESDA,MD. STANFORD UNIV,DEPT MICROBIOL & IMMUNOL,STANFORD,CA 94305. HOP PURPAN,CNRS,UPR 8291,TOULOUSE,FRANCE. UNIV FLORIDA,DEPT PATHOL,GAINESVILLE,FL 32611. CHILDRENS HOSP,RANGOS RES CTR,PITTSBURGH,PA 15213. UNIV TEXAS,MD ANDERSON CANCER CTR,DEPT EPIDEMIOL,HOUSTON,TX 77030. STANFORD UNIV,DEPT MED,STANFORD,CA 94305. FU NICHD NIH HHS [R01 HD 19469-06]; PHS HHS [28860, 23963] NR 44 TC 2 Z9 2 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA OSNEY MEAD, OXFORD, OXON, ENGLAND OX2 0EL SN 0960-7420 J9 EUR J IMMUNOGENET JI Eur. J. Immunogenet. PD OCT PY 1996 VL 23 IS 5 BP 361 EP 370 PG 10 WC Genetics & Heredity; Immunology SC Genetics & Heredity; Immunology GA VN275 UT WOS:A1996VN27500004 PM 8909943 ER PT J AU Montague, JW Cidlowski, JA AF Montague, JW Cidlowski, JA TI Cellular catabolism in apoptosis: DNA degradation and endonuclease activation SO EXPERIENTIA LA English DT Review DE apoptosis; cyclophilin; DNA degradation; NUC18; endonuclease ID CIS-TRANS-ISOMERASE; BINDING-PROTEIN CYCLOPHILIN; CALCIUM-DEPENDENT ENDONUCLEASE; YEAST CYCLOPHILIN; RAT THYMOCYTES; CYCLOSPORINE-A; DEOXYRIBONUCLEASE-II; GEL-ELECTROPHORESIS; NEUROSPORA-CRASSA; SECRETORY PATHWAY AB Recent research has focused on identifying the biochemical events associated with the apoptotic process. These include specific degradation of the chromatin which was described by Wyllie in 1980 [1], with the report of the appearance of discretely sized DNA fragments from apoptotic rat thymocytes. The fragments corresponded in size to strands of DNA that were cleaved at internucleosomal regions and create a 'ladder pattern' when electrophoresed on an agarose gel. Because of its near universality, internucleosomal DNA degradation is considered a diagnostic hallmark of cells undergoing apoptosis. It is of great interest to identify the enzymes involved, and some of the candidates will be discussed. C1 NIEHS,RES TRIANGLE PK,NC 27709. NR 56 TC 42 Z9 45 U1 0 U2 0 PU BIRKHAUSER VERLAG AG PI BASEL PA PO BOX 133 KLOSTERBERG 23, CH-4010 BASEL, SWITZERLAND SN 0014-4754 J9 EXPERIENTIA JI Experientia PD OCT PY 1996 VL 52 IS 10-11 BP 957 EP 962 DI 10.1007/BF01920104 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VQ942 UT WOS:A1996VQ94200005 PM 8917726 ER PT J AU Jones, BE Thompson, EW Hodos, W Waldbillig, RJ Chader, GJ AF Jones, BE Thompson, EW Hodos, W Waldbillig, RJ Chader, GJ TI Scleral matrix metalloproteinases, serine proteinase activity and hydrational capacity are increased in myopia induced by retinal image degradation SO EXPERIMENTAL EYE RESEARCH LA English DT Article DE eye; growth; myopia; sclera; metalloproteinase; serine protease; extracellular matrix; stromelysin; gelatinase ID HUMAN ARTICULAR-CARTILAGE; TISSUE INHIBITOR; INTERSTITIAL COLLAGENASE; INTERGLOBULAR DOMAIN; NEUTROPHIL ELASTASE; DEPRIVATION MYOPIA; CHICKS; IDENTIFICATION; AGGRECAN; GROWTH AB In the avian model of myopia, retinal image degradation quickly leads to ocular enlargement. We now give evidence that, regionally specific changes in ocular size are correlated with both biomechanical indices of scleral remodeling, e.g. hydration capacity and with biochemical changes in proteinase activities. The latter include a 72 kDa matrix metalloproteinase (putatively MMP-2), other gelatin-binding MMPs, an acid pH MMP and a serine protease. Specifically, we have found that increases in scleral hydrational capacity parallel increases in collagen degrading activities. Gelatin zymography reveals that eyes with 7 days of retinal image degradation have elevated levels (1.4-fold) of gelatinolytic activities at 72 and 67 kDa M(r) in equatorial and posterior pole regions of the sclera while, after 14 days of treatment, increases are no longer apparent. Lower M(r) zymographic activities at 50, 46 and 37 kDa M(r) are collectively increased in eyes treated for both 7 and 14 days (1.4- and 2.4-fold respectively) in the equator and posterior pole areas of enlarging eyes. Western blot analyses of scleral extracts with an antibody to human MMP-2 reveals immunoreactive bands at 65, 30 and 25 kDa. Zymograms incubated under slightly acidic conditions reveal that, in enlarging eyes, MMP activities at 25 and 28 kDa M(r) are increased in scleral equator and posterior pole (1.6- and 4.5-fold respectively). A TIMP-like protein is also identified in sclera and cornea by Western blot analysis. Finally, retinal-image degradation also increases (similar to 2.6-fold) the activity of a 23.5 kDa serine proteinase in limbus, equator and posterior pole sclera that is inhibited by aprotinin and soybean trypsin inhibitor. Taken together, these results indicate that eye growth induced by retinal-image degradation involves increases in the activities of multiple scleral proteinases that could modify the biomechanical properties of scleral structural components and contribute to tissue remodeling and growth. (C) 1996 Academic Press Limited C1 NEI,RETINAL CELL & MOL BIOL LAB,NIH,BETHESDA,MD 20892. GEORGETOWN UNIV,SCH MED,DEPT CELL BIOL,WASHINGTON,DC 20007. GEORGETOWN UNIV,SCH MED,VINCENT T LOMBARDI CANC RES CTR,WASHINGTON,DC 20007. GEORGETOWN UNIV,SCH MED,DEPT ORTHOPED SURG,WASHINGTON,DC 20007. UNIV MARYLAND,DEPT PSYCHOL,COLLEGE PK,MD 20742. RI Thompson, Erik/A-1425-2009 OI Thompson, Erik/0000-0002-9723-4924 FU NEI NIH HHS [EY-04742] NR 36 TC 27 Z9 32 U1 0 U2 4 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON, ENGLAND NW1 7DX SN 0014-4835 J9 EXP EYE RES JI Exp. Eye Res. PD OCT PY 1996 VL 63 IS 4 BP 369 EP 381 DI 10.1006/exer.1996.0127 PG 13 WC Ophthalmology SC Ophthalmology GA VQ440 UT WOS:A1996VQ44000003 PM 8944544 ER PT J AU Kamiya, T Zigler, JS AF Kamiya, T Zigler, JS TI Long-term maintenance of monkey lenses in organ culture: A potential model system for the study of human cataractogenesis SO EXPERIMENTAL EYE RESEARCH LA English DT Article DE Rhesus monkey; lens; organ culture; crystallin synthesis; xylose; cataract; diabetes ID REGIONAL DISTRIBUTION; OXIDATIVE STRESS; RAT LENSES; BINDING; QUANTITATION; EPITHELIUM; PROTEINS; INVITRO AB Lenses from Rhesus monkeys were maintained in protein-free medium and both their biochemical and histological changes were examined. Lenses were dissected under a dissecting microscope to ensure minimal contamination by other ocular tissue, which allowed successful observation of whole lens morphology during culture, Lenses in culture showed no acute leakage of protein into medium, indicating that they had not been physically damaged during dissection. Cultured lenses remained transparent for 21 days without any noticeable histological changes. Even at 21 days of culture, S-35- methionine incorporation into lens protein was approximately 60% of the level observed in fresh lenses. The amounts of most protein species were rather constant in the lens capsule/epithelium throughout culture, but the amount of crystallins gradually decreased during the first 14 days of culture due to selective decrease in crystallin synthesis. After day 14, both the amount of crystallins and their synthesis became stable. When 30 mM fructose, present in the control medium, was replaced with 30 mM xylose, lens opacification with some histological abnormalities such as vacuole formation was observed. In the xylose-induced cataract, the protein synthetic activity decreased dramatically. This system would be a valuable tool in investigating the mechanisms of lens homeostasis as well as mechanisms of cataractogenesis in primate lens. (C) 1996 Academic Press Limited C1 NEI,LAB MECHANISMS OCULAR DIS,NIH,BETHESDA,MD 20892. NR 23 TC 7 Z9 7 U1 0 U2 0 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON, ENGLAND NW1 7DX SN 0014-4835 J9 EXP EYE RES JI Exp. Eye Res. PD OCT PY 1996 VL 63 IS 4 BP 425 EP 431 DI 10.1006/exer.1996.0132 PG 7 WC Ophthalmology SC Ophthalmology GA VQ440 UT WOS:A1996VQ44000008 PM 8944549 ER PT J AU Korn, WM Weghuis, DEMO Suijkerbuijk, RF Schmidt, U Otto, T duManoir, S vanKessel, AG Harstrick, A Seeber, S Becher, R AF Korn, WM Weghuis, DEMO Suijkerbuijk, RF Schmidt, U Otto, T duManoir, S vanKessel, AG Harstrick, A Seeber, S Becher, R TI Defection of chromosomal DNA gains and losses in testicular germ cell tumors by comparative genomic hybridization SO GENES CHROMOSOMES & CANCER LA English DT Article ID MOLECULAR CYTOGENETIC ANALYSIS; INSITU HYBRIDIZATION; T-COMPLEX; EXPRESSION; GENE; IDENTIFICATION; AMPLIFICATION; HETEROZYGOSITY; PROTOONCOGENES; SEQUENCES AB To extend the results of conventional cytogenetic analysis of testicular germ cell tumors (TGCTs), we applied the new molecular cytogenetic method of comparative genomic hybridization (CGH), which enables the detection of chromosomal imbalances without the need for dividing cells. DNA from I I TGCTs was studied by CGH. In all rumors examined, gain of 12p, mostly of the whole p arm, could be demonstrated. However, in three tumors, an amplification of 12p material restricted to the chromosomal bands 12p11.2-p12.1 was found. Further fluorescence in situ hybridization (FISH) analysis using a yeast artificial chromosome (YAC) that was previously mapped to that region revealed multiple copies of that chromosomal segment in interphase nuclei of these tumors. This finding is an important clue to the localization of candidate protooncogenes at 12p involved in TGCTs. Gains of small chromosomal regions at 2p, 4q, 6p, and 19p were also detected recurrently. Furthermore, gains of chromosomes 8, 14, 21, and X as well as loss of chromosome 13 were frequent findings. In conclusion, CGH provides new insights into genetic alterations of TGCTs. By using CGH, chromosomal subregions could be identified that may harbor genes involved in the pathogenesis of this malignancy. (C) 1996 Wiley-Liss, Inc. C1 UNIV ESSEN GESAMTHSCH,SCH MED,W GERMAN CANC CTR,ESSEN,GERMANY. UNIV NIJMEGEN HOSP,DEPT HUMAN GENET,NIJMEGEN,NETHERLANDS. NIH,BETHESDA,MD 20892. RI Geurts van Kessel, Ad/A-2810-2010; Olde Weghuis, Danielle/C-4674-2016 NR 38 TC 75 Z9 75 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC 605 THIRD AVE, NEW YORK, NY 10158-0012 SN 1045-2257 J9 GENE CHROMOSOME CANC JI Gene Chromosomes Cancer PD OCT PY 1996 VL 17 IS 2 BP 78 EP 87 DI 10.1002/(SICI)1098-2264(199610)17:2<78::AID-GCC2>3.0.CO;2-Y PG 10 WC Oncology; Genetics & Heredity SC Oncology; Genetics & Heredity GA VP824 UT WOS:A1996VP82400002 PM 8913724 ER PT J AU Mushegian, AR Koonin, EV AF Mushegian, AR Koonin, EV TI Sequence analysis of eukaryotic developmental proteins: Ancient and novel domains SO GENETICS LA English DT Review ID HOMEODOMAIN-DNA COMPLEX; PROGRAMMED CELL-DEATH; HELIX-TURN-HELIX; CRYSTAL-STRUCTURE; BINDING-PROTEINS; PROTOONCOGENE BCL-2; SECONDARY STRUCTURE; FLOWER DEVELOPMENT; CONSERVED REGIONS; MOLECULAR-BIOLOGY AB Most of the genes involved in the development of multicellular eukaryotes encode large, multidomain proteins. To decipher the major trends in the evolution of these proteins and make functional predictions for uncharacterized domains, we applied a strategy of sequence database search that includes construction of specialized data sets and iterative subsequence masking. This computational approach allowed us to detect previously unnoticed but potentially important sequence similarities. Developmental gene products are enriched in predicted nonglobular regions as compared to unbiased sets of eukaryotic and bacterial proteins. Developmental genes that act intracellularly, primarily at the level of transcription regulation, typically code for proteins containing highly conserved DNA-binding domains, most of which appear to have evolved before the radiation of bacteria and eukaryotes. We identified bacterial homologues, namely a protein family that includes the Escherichia coli universal stress protein UspA, for the MADS-box transcription regulators previously described only in eukaryotes. We also show that the FUS6 family of eukaryotic proteins contains a putative DNA-binding domain related to bacterial helix-turn-helix transcription regulators. Developmental proteins that act extracellularly are less conserved and often do not have bacterial homologues. Nevertheless, several provocative similarities between different groups of such proteins were detected. RP Mushegian, AR (reprint author), NIH,NATL LIB MED,NATL CTR BIOTECHNOL INFORMAT,BLDG 38A,8600 ROCKVILLE PIKE,BETHESDA,MD 20894, USA. NR 103 TC 39 Z9 42 U1 1 U2 1 PU GENETICS PI BALTIMORE PA 428 EAST PRESTON ST, BALTIMORE, MD 21202 SN 0016-6731 J9 GENETICS JI Genetics PD OCT PY 1996 VL 144 IS 2 BP 817 EP 828 PG 12 WC Genetics & Heredity SC Genetics & Heredity GA VK261 UT WOS:A1996VK26100033 PM 8889542 ER PT J AU Mock, BA Liu, LM LePaslier, D Huang, S AF Mock, BA Liu, LM LePaslier, D Huang, S TI The B-lymphocyte maturation promoting transcription factor BLIMP1/PRDI-BF1 maps to D6S447 on human chromosome 6q21-q22.1 and the syntenic region of mouse chromosome 10 SO GENOMICS LA English DT Article ID NON-HODGKINS-LYMPHOMA; ZINC-FINGER PROTEIN; 2 DISTINCT REGIONS; CELL-LINES; GENE; MELANOMA; LEUKEMIA; TRANSLOCATIONS; KARYOTYPES; ACTIVATION AB The human PRDI-BF1 or BLIMP1 gene and its mouse homolog Blimp1 are members of the recently realized PR domain family that includes the retinoblastoma interacting zinc finger gene RIZ and the MDS1-EVI1 leukemia cancer gene. The specific high-level expression of Blimp1 in late B and plasma cells, its induction during B-cell differentiation, and its ability to drive B-cell maturation suggest that this gene may play a role in the differentiation and pathogenesis of B cells. We have now mapped the physical location of BLIMP1 near the marker D6S447 on human chromosome 6q21-q22.1; we have also mapped Blimp1 to mouse Chromosome 10 at 14 cM distal to the Myb locus and to a region homologous to the location of BLIMP1. Deletions of the 6q21-q22 region are common in several human malignancies, particularly in B-cell non-Hodgkin lymphoma (B-NRL). The data led us to suggest that BLIMP1 may be a candidate B-NHL tumor suppressor gene. (C) 1996 Academic Press, Inc. C1 LA JOLLA CANC RES CTR,BURNHAM INST,LA JOLLA,CA 92037. NCI,NIH,BETHESDA,MD 20892. FDN JEAN DAUSSET,CTR ETUD POLYMORPHISME HUMAIN,F-75010 PARIS,FRANCE. FU NCI NIH HHS [N01-BC-21075, R01CA57496] NR 29 TC 37 Z9 42 U1 0 U2 0 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0888-7543 J9 GENOMICS JI Genomics PD OCT 1 PY 1996 VL 37 IS 1 BP 24 EP 28 DI 10.1006/geno.1996.0516 PG 5 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA VL371 UT WOS:A1996VL37100004 PM 8921366 ER PT J AU Riedy, MC Dutra, AS Blake, TB Modi, W Lal, BK Davis, J Bosse, A OShea, JJ Johnston, JA AF Riedy, MC Dutra, AS Blake, TB Modi, W Lal, BK Davis, J Bosse, A OShea, JJ Johnston, JA TI Genomic sequence, organization, and chromosomal localization of human JAK3 SO GENOMICS LA English DT Article ID SEVERE COMBINED IMMUNODEFICIENCY; CYTOKINE RECEPTOR SUPERFAMILY; PROTEIN-TYROSINE KINASES; JANUS-KINASE; GAMMA-CHAIN; INTERLEUKIN-2; CELLS; MUTATION; FAMILY; PHOSPHORYLATION AB Members of the Janus (JAK) protein tyrosine kinase family including JAK3 have recently emerged as important components in cytokine signal transduction. Mutations of JAK3 have been found in a number of patients who present with severe combined immunodeficiency. To facilitate the further identification of JAK3-SCID patients and to understand the structure of JAK3 better, we undertook the determination of the genomic sequence, organization, and chromosomal localization of the JAK3 gene, The JAK3 gene was found to consist of 19 exons and 18 introns. Interestingly, the organization of the kinase-(JH1) and pseudokinase-(JH2) domains were found to be dissimilar. In addition, the JAK3 gene was localized to human chromosome 19p13.1. These data should facilitate the identification of patients with this new form of immunodeficiency and will provide insight into the structure of this kinase. (C) 1996 Academic Press, Inc. C1 NIH,NCHGR,BETHESDA,MD 20892. NCI,FREDERICK CANC RES & DEV CTR,ABL,FREDERICK,MD 21702. RP Riedy, MC (reprint author), NIAMS,LYMPHOCYTE CELL BIOL SECT,ARTHRIT & RHEUMATISM BRANCH,BETHESDA,MD 20892, USA. NR 33 TC 32 Z9 34 U1 0 U2 1 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0888-7543 J9 GENOMICS JI Genomics PD OCT 1 PY 1996 VL 37 IS 1 BP 57 EP 61 DI 10.1006/geno.1996.0520 PG 5 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA VL371 UT WOS:A1996VL37100008 PM 8921370 ER PT J AU Usdin, TB Modi, W Bonner, TI AF Usdin, TB Modi, W Bonner, TI TI Assignment of the human PTH2 receptor gene (PTHR2) to chromosome 2q33 by fluorescence in situ hybridization SO GENOMICS LA English DT Article ID PARATHYROID-HORMONE; LINKAGE C1 FREDERICK CANC RES & DEV FACIL,FREDERICK,MD. RP Usdin, TB (reprint author), NIMH,CELL BIOL LAB,36 CONVENT DR MSC4090,BETHESDA,MD 20892, USA. NR 12 TC 13 Z9 14 U1 0 U2 0 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0888-7543 J9 GENOMICS JI Genomics PD OCT 1 PY 1996 VL 37 IS 1 BP 140 EP 141 DI 10.1006/geno.1996.0532 PG 2 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA VL371 UT WOS:A1996VL37100020 PM 8921382 ER PT J AU Orvisky, E VonKollmar, DE Melton, DJ Stone, AL AF Orvisky, E VonKollmar, DE Melton, DJ Stone, AL TI Methods of analysis of heparin-mimetic sulfated xylans by high-performance gel permeation chromatography, polyacrylamide-gel electrophoresis and polyanion-induced metachromacy SO GLYCOBIOLOGY LA English DT Meeting Abstract C1 NIMH,NATL INST HLTH,BETHESDA,MD 20892. NR 1 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS UNITED KINGDOM PI OXFORD PA WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP SN 0959-6658 J9 GLYCOBIOLOGY JI Glycobiology PD OCT PY 1996 VL 6 IS 7 BP 124 EP 124 PG 1 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VT565 UT WOS:A1996VT56500096 ER PT J AU Stone, AL Melton, J Horne, MK AF Stone, AL Melton, J Horne, MK TI Structural basis underlying the heparin-mimetic capacity of sulfated xylans in anticoagulation in vitro SO GLYCOBIOLOGY LA English DT Meeting Abstract C1 NIH,BETHESDA,MD 20892. NICHHD,IRP,BETHESDA,MD 20892. CPD,CC,BETHESDA,MD 20892. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS UNITED KINGDOM PI OXFORD PA WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP SN 0959-6658 J9 GLYCOBIOLOGY JI Glycobiology PD OCT PY 1996 VL 6 IS 7 BP 1607 EP 1607 PG 1 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VT565 UT WOS:A1996VT56500225 ER PT J AU Platt, FM Neises, GR Proia, RL Dwek, RA Butters, TD AF Platt, FM Neises, GR Proia, RL Dwek, RA Butters, TD TI Novel inhibitors of glycosphingolipid biosynthesis SO GLYCOBIOLOGY LA English DT Meeting Abstract C1 MONSANTO CO,ST LOUIS,MO 63198. UNIV OXFORD,DEPT BIOCHEM,GLYCOBIOL INST,OXFORD OX1 3QU,ENGLAND. NIDDKD,NATL INST HLTH,BETHESDA,MD 20892. RI Platt, Frances/G-1004-2010; Proia, Richard/A-7908-2012 NR 0 TC 1 Z9 1 U1 0 U2 0 PU OXFORD UNIV PRESS UNITED KINGDOM PI OXFORD PA WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP SN 0959-6658 J9 GLYCOBIOLOGY JI Glycobiology PD OCT PY 1996 VL 6 IS 7 BP E3 EP E3 PG 1 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VT565 UT WOS:A1996VT56500034 ER PT J AU Hayashi, S GuexCrosier, Y Delvaux, A Velu, T Roberge, FG AF Hayashi, S GuexCrosier, Y Delvaux, A Velu, T Roberge, FG TI Interleukin 10 inhibits inflammatory cells infiltration in endotoxin-induced uveitis SO GRAEFES ARCHIVE FOR CLINICAL AND EXPERIMENTAL OPHTHALMOLOGY LA English DT Article ID TUMOR-NECROSIS-FACTOR; NITRIC-OXIDE SYNTHASE; MESSENGER-RNA LEVELS; INTERFERON-GAMMA; RAT; IL-10; MACROPHAGES; INDUCTION; MOUSE; BETA AB Background: Endotoxin-induced uveitis (EIU) is a model for acute anterior uveitis associated with a variety of pro-inflammatory cytokines and nitric oxide production. Interleukin 10 (IL-10) downregulates these inflammatory mediators. We report a study of the effect of systemic administration of IL-10 on the inflammatory parameters of EIU. Methods. Uveitis was induced in C3H/HeN mice by subcutaneous injection of 200 mu g lipopolysaccharide (LPS) per mouse. Intraocular inflammation was assessed by leukocyte count and measurement of the protein concentration in the aqueous humor (AH). Mouse recombinant IL-10 at 1000 U or its vehicle alone were administered by three intravenous injections given 4.0 h and 0.5 h before and 8.0 h after LPS injection. Results: The inflammatory cell infiltration in the eyes was significantly reduced in four of five experiments from 40% to 64% in the groups treated with IL-10 compared to the control groups (P<0.05). In contrast, the level of protein exudation in the anterior chamber (AC) was not significantly affected by IL-10 treatment. Conclusion: IL-10 reduces the cellular infiltration in the ocular inflammation produced by endotoxin. This result suggests potential usefulness for IL-10 in the treatment of severe anterior uveitis with a strong cellular component. C1 NEI,NATL INST HLTH,BETHESDA,MD 20892. FREE UNIV BRUSSELS,INST RECH INTERDISCIPLINAIRE,B-1070 BRUSSELS,BELGIUM. FREE UNIV BRUSSELS,ERASME HOSP,INST RECH INTERDISCIPLINAIRE,B-1070 BRUSSELS,BELGIUM. FREE UNIV BRUSSELS,ERASME HOSP,DEPT MED GENET,B-1070 BRUSSELS,BELGIUM. NR 23 TC 26 Z9 29 U1 0 U2 0 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 SN 0721-832X J9 GRAEF ARCH CLIN EXP JI Graefes Arch. Clin. Exp. Ophthalmol. PD OCT PY 1996 VL 234 IS 10 BP 633 EP 636 DI 10.1007/BF00185297 PG 4 WC Ophthalmology SC Ophthalmology GA VM741 UT WOS:A1996VM74100008 PM 8897056 ER PT J AU Tiberghien, P Cahn, JY Ferrand, C Milpied, N Contassot, E Angonin, R Sallard, D Robinet, E Reynolds, CW Certoux, JM Rousseau, E Jacob, W Herve, P AF Tiberghien, P Cahn, JY Ferrand, C Milpied, N Contassot, E Angonin, R Sallard, D Robinet, E Reynolds, CW Certoux, JM Rousseau, E Jacob, W Herve, P TI Use of the thymidine-kinase gene of the herpes simplex 1 virus as therapeutic tool in transplantation of blood stem cells SO HEMATOLOGY AND CELL THERAPY LA French DT Article; Proceedings Paper CT 5th Cell Therapy Workshop CY OCT 18, 1996 CL LYON, FRANCE C1 CHU BESANCON,ETS FRANCHE COMTE,SERV ANAT PATHOL,F-25030 BESANCON,FRANCE. CHU NANTES,SERV HEMATOL,F-44035 NANTES 01,FRANCE. NCI,FREDERICK CANC RES & DEV CTR,FREDERICK,MD. GENET THERAPY INC,GAITHERSBURG,MD. RP Tiberghien, P (reprint author), CHU BESANCON,ETS FRANCHE COMTE,SERV HEMATOL,F-25030 BESANCON,FRANCE. NR 0 TC 0 Z9 0 U1 0 U2 0 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 SN 1430-2772 J9 HEMATOL CELL THER JI Hematol. Cell Ther. PD OCT PY 1996 VL 38 IS 5 BP 464 EP 464 DI 10.1007/s00282-996-0464-3 PG 1 WC Oncology; Hematology SC Oncology; Hematology GA VP204 UT WOS:A1996VP20400015 PM 9044823 ER PT J AU Biggar, RJ Rabkin, CS AF Biggar, RJ Rabkin, CS TI The epidemiology of AIDS-related neoplasms SO HEMATOLOGY-ONCOLOGY CLINICS OF NORTH AMERICA LA English DT Article ID ACQUIRED-IMMUNODEFICIENCY-SYNDROME; SARCOMA-ASSOCIATED HERPESVIRUS; NON-HODGKINS-LYMPHOMA; EPSTEIN-BARR VIRUS; KAPOSIS-SARCOMA; HIV-INFECTION; DNA-SEQUENCES; RENAL-TRANSPLANTATION; HOMOSEXUAL MEN; UNITED-STATES AB The magnitude for and risk factors of the two most important AIDS neoplasms, Kaposi's sarcoma and non-Hodgkin's lymphoma, are reviewed in detail. The association between AIDS and other cancers is mostly speculative because surveillance biases tend to favor detecting associations that may be spurious. The overall relative risk of other cancers appears, however, to be only twofold above that in the general population, with associations being most convincing for anal (but not cervical) cancer and leiomyosarcoma and possible also for Hodgkin's disease, testicular cancer, and conjunctival cancers. RP Biggar, RJ (reprint author), NCI,VIRAL EPIDEMIOL BRANCH,6130 EXECUT BLVD,EPN-434,ROCKVILLE,MD 20852, USA. NR 79 TC 114 Z9 115 U1 0 U2 1 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0889-8588 J9 HEMATOL ONCOL CLIN N JI Hematol. Oncol. Clin. North Am. PD OCT PY 1996 VL 10 IS 5 BP 997 EP & DI 10.1016/S0889-8588(05)70380-4 PG 15 WC Oncology; Hematology SC Oncology; Hematology GA VG731 UT WOS:A1996VG73100002 PM 8880192 ER PT J AU Karp, JE Pluda, JM Yarchoan, R AF Karp, JE Pluda, JM Yarchoan, R TI AIDS-related Kaposi's sarcoma - A template for the translation of molecular pathogenesis and targeted therapeutic approaches SO HEMATOLOGY-ONCOLOGY CLINICS OF NORTH AMERICA LA English DT Review ID HUMAN-IMMUNODEFICIENCY-VIRUS; FIBROBLAST GROWTH-FACTOR; COLONY-STIMULATING FACTOR; TRANS-RETINOIC ACID; ENDOTHELIAL-CELL PROLIFERATION; PENTOSAN POLYSULFATE PPS; NECROSIS-FACTOR RECEPTOR; ANGIOGENESIS IN-VIVO; TYPE-1 TAT PROTEIN; ONCOSTATIN-M AB AIDS-related Kaposi's sarcoma (KS) represents a complex interaction of host and viral factors. There are a number of fundamental questions surrounding the interplay between the disparate factors that can contribute to the pathogenesis and pathophysiology of this disease. Targets such as the enhancement of immune function, inhibition of angiogenic factors or immunostimulatory cytokines, inhibition of viral proteins such as Tat, or hormonal manipulations are now or will in the future become the focus of research to develop innovative anti-KS therapy and prevention measures. Antiviral approaches aimed at HIV or other viruses may potentially target a number of steps in KS pathogenesis. This article reviews diverse modalities-cytotoxic, antiviral, gene-directed, growth factor-targeted, and antiangiogenesis-that singly, or more likely in combination, stand to make an impact on the cure and prevention of AIDS-related KS. C1 NCI,CHEMOPREVENT BRANCH,DIV CANC,BETHESDA,MD 20892. NCI,INVESTIGAT DRUG BRANCH,CLIN TRIALS EVALUAT PROGRAM,DIV CANC TREATMENT DIAG & CTR,BETHESDA,MD 20892. NCI,RETROVIRAL DIS SECT,MED BRANCH,DIV CLIN SCI,BETHESDA,MD 20892. NR 131 TC 22 Z9 23 U1 0 U2 2 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0889-8588 J9 HEMATOL ONCOL CLIN N JI Hematol. Oncol. Clin. North Am. PD OCT PY 1996 VL 10 IS 5 BP 1031 EP & DI 10.1016/S0889-8588(05)70383-X PG 20 WC Oncology; Hematology SC Oncology; Hematology GA VG731 UT WOS:A1996VG73100005 PM 8880195 ER PT J AU Mittur, A Kaplowitz, N Kempner, E Ookhtens, M AF Mittur, A Kaplowitz, N Kempner, E Ookhtens, M TI Novel features of the canalicular GSH transporter revealed by radiation target size analysis. SO HEPATOLOGY LA English DT Meeting Abstract C1 UNIV SO CALIF,DEPT MED,LOS ANGELES,CA. UNIV SO CALIF,DEPT BIOMED ENGN,LOS ANGELES,CA 90089. NIH,BETHESDA,MD 20892. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 20 EP 20 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500020 ER PT J AU Heller, T Purcell, RH Klimpel, KR Emerson, SU AF Heller, T Purcell, RH Klimpel, KR Emerson, SU TI Assessment of the function of the third open reading frame protein of the hepatitis E virus SO HEPATOLOGY LA English DT Meeting Abstract C1 NIH,BETHESDA,MD 20892. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 248 EP 248 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500248 ER PT J AU Hoofnagle, JH Lombardero, M Wei, Y Yun, AJ Yang, L Kim, JP AF Hoofnagle, JH Lombardero, M Wei, Y Yun, AJ Yang, L Kim, JP TI Hepatitis G virus (HGV) infection before and after liver transplantation for fulminant hepatic failure (FHF) and cryptogenic cirrhosis. SO HEPATOLOGY LA English DT Meeting Abstract C1 NIH,BETHESDA,MD 20892. UNIV PITTSBURGH,DEPT EPIDEMIOL,PITTSBURGH,PA 15261. GENELABS TECHNOL INC,REDWOOD CITY,CA. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 250 EP 250 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500251 ER PT J AU Marrone, A Shih, JWK Nakatsuji, Y Alter, HJ Lau, D Vergalla, J Hoffnagle, JH AF Marrone, A Shih, JWK Nakatsuji, Y Alter, HJ Lau, D Vergalla, J Hoffnagle, JH TI Hepatitis G virus (HGV) among patients with chronic hepatitis B. SO HEPATOLOGY LA English DT Meeting Abstract C1 NIH,LIVER DIS SECT,BETHESDA,MD 20892. NIH,DIV TRANSFUS MED,BETHESDA,MD 20892. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 393 EP 393 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500393 ER PT J AU Marrone, A Shih, JWK Nakatsuji, Y Alter, HJ Lau, D Herion, D Vergalla, J Hoofnagle, JH AF Marrone, A Shih, JWK Nakatsuji, Y Alter, HJ Lau, D Herion, D Vergalla, J Hoofnagle, JH TI Lack of effect of ribavirin on serum levels of hepatitis C virus (HCV) and hepatitis G virus (HGV) RNA. SO HEPATOLOGY LA English DT Meeting Abstract C1 NIH,LIVER DIS SECT,BETHESDA,MD 20892. NIH,DIV TRANSFUS MED,BETHESDA,MD 20892. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 394 EP 394 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500395 ER PT J AU Tanaka, E Alter, HJ Nakatsuji, Y Shih, JWK Kim, JP Matsumoto, A Kobayashi, M Kiyosawa, K AF Tanaka, E Alter, HJ Nakatsuji, Y Shih, JWK Kim, JP Matsumoto, A Kobayashi, M Kiyosawa, K TI Effect of hepatitis G virus co-infection on patients with chronic hepatitis C. SO HEPATOLOGY LA English DT Meeting Abstract C1 SHINSHU UNIV,SCH MED,DEPT INTERNAL MED 2,MATSUMOTO,NAGANO 390,JAPAN. NIH,DEPT TRANSFUS MED,BETHESDA,MD 20892. GENELABS TECHNOL INC,REDWOOD CITY,CA. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 400 EP 400 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500400 ER PT J AU Nakatsuji, Y Shih, JWK Kobayashi, M Tanaka, E Kiyosawa, K Kim, JP Alter, HJ AF Nakatsuji, Y Shih, JWK Kobayashi, M Tanaka, E Kiyosawa, K Kim, JP Alter, HJ TI Hepatitis G virus (HGV) infection in Japanese patients infected with hepatitis C virus (HCV). SO HEPATOLOGY LA English DT Meeting Abstract C1 NIH,DEPT TRANSFUS MED,BETHESDA,MD 20892. SHINSHU UNIV,MATSUMOTO,NAGANO 390,JAPAN. GENELABS TECHNOL INC,REDWOOD CITY,CA. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 403 EP 403 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500402 ER PT J AU Baumert, TF Ito, S Wong, DT Greenberg, HB Liang, TJ AF Baumert, TF Ito, S Wong, DT Greenberg, HB Liang, TJ TI Synthesis of hepatitis C virus like particles in insect cells. SO HEPATOLOGY LA English DT Meeting Abstract C1 MASSACHUSETTS GEN HOSP,GI UNIT,BOSTON,MA 02114. NIH,LIVER DIS SECT,BETHESDA,MD. HARVARD UNIV,SCH MED,ELECTRON MICROSCOPY FACIL,BOSTON,MA. STANFORD UNIV,SCH MED,PALO ALTO,CA 94304. NR 0 TC 0 Z9 0 U1 0 U2 1 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 500 EP 500 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500499 ER PT J AU Omori, N Omori, M Evarts, RP Teramoto, T Thorgeirsson, SS AF Omori, N Omori, M Evarts, RP Teramoto, T Thorgeirsson, SS TI Compensatory effect of the FLT-3 ligand/receptor system for SCF/c-kit in bile duct ligated SL and W mice. SO HEPATOLOGY LA English DT Meeting Abstract C1 NCI,EXPT CARCINOGENESIS LAB,NIH,BETHESDA,MD 20892. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 504 EP 504 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500503 ER PT J AU Preisegger, KH Factor, V Stumptner, C Denk, H Thorgeirsson, SS AF Preisegger, KH Factor, V Stumptner, C Denk, H Thorgeirsson, SS TI TGF-beta inhibits the proliferation of the potential liver stem cell compartment in the mouse SO HEPATOLOGY LA English DT Meeting Abstract C1 NIH,EXPT CARCINOGENESIS LAB,BETHESDA,MD 20892. GRAZ UNIV,DEPT PATHOL,A-8010 GRAZ,AUSTRIA. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 518 EP 518 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500518 ER PT J AU Sakata, H Takayama, H Sharp, R Taylor, W Kriebel, P Rubin, JS Merlino, G LaRochelle, W AF Sakata, H Takayama, H Sharp, R Taylor, W Kriebel, P Rubin, JS Merlino, G LaRochelle, W TI Hepatocyte growth factor scatter factor overexpression alters liver growth and development in transgenic mice. SO HEPATOLOGY LA English DT Meeting Abstract C1 NCI,CELLULAR & MOL BIOL LAB,BETHESDA,MD 20892. NCI,MOL BIOL LAB,BETHESDA,MD 20892. GUNMA UNIV,SCH MED,DEPT MED 1,GUNMA,JAPAN. NR 0 TC 0 Z9 0 U1 0 U2 1 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 535 EP 535 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500533 ER PT J AU Kawamura, T Furusaka, A Kozie, MJ Wang, TC Liang, TJ Schmidt, EV AF Kawamura, T Furusaka, A Kozie, MJ Wang, TC Liang, TJ Schmidt, EV TI Transgenic expression of hepatitis C virus structural proteins. SO HEPATOLOGY LA English DT Meeting Abstract C1 MASSACHUSETTS GEN HOSP,CTR CANC,CHARLESTOWN,MA. MASSACHUSETTS GEN HOSP,DEPT MED,GASTROINTESTINAL UNIT,BOSTON,MA. HARVARD UNIV,SCH MED,BOSTON,MA. NIDDK,LIVER DIS SECT,NIH,BETHESDA,MD. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 547 EP 547 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500545 ER PT J AU Lau, D Comanor, L Minor, JM Watson, G Hoofnagle, JH AF Lau, D Comanor, L Minor, JM Watson, G Hoofnagle, JH TI Statistical models for predicting a response to alpha interferon in patients with chronic hepatitis B. SO HEPATOLOGY LA English DT Meeting Abstract C1 NIDDK,LIVER DIS SECT,NIH,BETHESDA,MD. CHIRON CORP,EMERYVILLE,CA 94608. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 611 EP 611 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500609 ER PT J AU Bukh, J Wantzin, P Krogsgaard, K Apgar, C Knudsen, F Purcell, RH AF Bukh, J Wantzin, P Krogsgaard, K Apgar, C Knudsen, F Purcell, RH TI Molecular study of GBV-C in dialysis patients. SO HEPATOLOGY LA English DT Meeting Abstract C1 NIAID,HEPATITIS VIRUSES SECT,LID,NIH,BETHESDA,MD. HVIDOVRE UNIV HOSP,DEPT CLIN IMMUNOL,COPENHAGEN,DENMARK. HVIDOVRE UNIV HOSP,DEPT NEPHROL,COPENHAGEN,DENMARK. HVIDOVRE UNIV HOSP,DEPT INFECT DIS,COPENHAGEN,DENMARK. RIGSHOSP,DK-2100 COPENHAGEN,DENMARK. HERLEV HOSP,COPENHAGEN,DENMARK. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 645 EP 645 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500644 ER PT J AU Forns, X Bukh, J Costa, J Olmedo, E Darnell, A Rodes, J Purcell, RH AF Forns, X Bukh, J Costa, J Olmedo, E Darnell, A Rodes, J Purcell, RH TI Detection of GBV-C RNA in RT-PCR assays with primers derived from different genomic regions. SO HEPATOLOGY LA English DT Meeting Abstract C1 HOSP CLIN BARCELONA,LIVER UNIT,BARCELONA,SPAIN. HOSP CLIN BARCELONA,DEPT NEPHROL,BARCELONA,SPAIN. NIAID,HEPATITIS VIRUSES SECT,NIH,LID,BETHESDA,MD 20892. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 654 EP 654 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500654 ER PT J AU Munoz, S Alter, H Liang, TJ Nakatsuji, Y Shih, J Reddy, R Jeffers, L Reid, A Marrone, A Rothstein, K Manzarbeitia, C AF Munoz, S Alter, H Liang, TJ Nakatsuji, Y Shih, J Reddy, R Jeffers, L Reid, A Marrone, A Rothstein, K Manzarbeitia, C TI Hepatitis G virus is present in serum of patients with fulminant hepatitis of unknown etiology. SO HEPATOLOGY LA English DT Meeting Abstract C1 ALBERT EINSTEIN MED CTR,CTR LIVER DIS,PHILADELPHIA,PA 19141. NIH,CTR CLIN,DEPT TRANSFUS MED,BETHESDA,MD 20892. NIH,LIVER DIS SECT,BETHESDA,MD 20892. MASSACHUSETTS GEN HOSP,GI UNIT,BOSTON,MA 02114. UNIV MIAMI,MIAMI,FL 33152. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 665 EP 665 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500664 ER PT J AU Lake, JR Lombardero, MS AF Lake, JR Lombardero, MS TI Evaluating patients with alcoholic liver disease (ALD) for liver transplantation (LT): Differences in clinical status and contraindications (CI). SO HEPATOLOGY LA English DT Meeting Abstract C1 UNIV CALIF SAN FRANCISCO,NIDDK,LIVER TRANSPLANTAT DATABASE LTD,SAN FRANCISCO,CA 94143. UNIV PITTSBURGH,DEPT EPIDEMIOL,PITTSBURGH,PA 15261. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 697 EP 697 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500696 ER PT J AU Farci, P Melpolder, JC Shimoda, A Peddis, G Strazzera, A Munoz, S Purcell, RH Alter, HJ AF Farci, P Melpolder, JC Shimoda, A Peddis, G Strazzera, A Munoz, S Purcell, RH Alter, HJ TI Studies of HCV quasispecies in patients with acute resolving hepatitis compared to those who progress to chronic hepatitis. SO HEPATOLOGY LA English DT Meeting Abstract C1 NIAID,NIH,BETHESDA,MD. UNIV CAGLIARI,DEPT INTERNAL MED,CAGLIARI,ITALY. ALBERT EINSTEIN MED CTR,PHILADELPHIA,PA 19141. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 895 EP 895 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28500893 ER PT J AU Baumert, TF Liang, TJ AF Baumert, TF Liang, TJ TI A novel HBV genetic element important for viral replication is defined by mutations in the core promotor region. SO HEPATOLOGY LA English DT Meeting Abstract C1 HARVARD UNIV,MASSACHUSETTS GEN HOSP,SCH MED,DEPT MED,GI UNIT,BOSTON,MA. NATL INST HLTH,LIVER DIS SECT,BETHESDA,MD. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 1120 EP 1120 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28501119 ER PT J AU Bukh, J Kim, J Govindarajan, S Apgar, C Wages, J Yun, A Piatak, M Purcell, RH AF Bukh, J Kim, J Govindarajan, S Apgar, C Wages, J Yun, A Piatak, M Purcell, RH TI Natural history of hepatitis G virus in chimpanzees. SO HEPATOLOGY LA English DT Meeting Abstract C1 NIAID,HEPATITIS VIRUSES SECT,LID,NIH,BETHESDA,MD 20892. GENELABS TECHNOL INC,REDWOOD CITY,CA. UNIV SO CALIF,RANCHO LOS AMIGOS HOSP,DOWNEY,CA 90242. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 1146 EP 1146 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28501146 ER PT J AU Iimuro, Y Kono, H Conner, HD Bradford, BU Mason, RP Wall, CA Thurman, RG AF Iimuro, Y Kono, H Conner, HD Bradford, BU Mason, RP Wall, CA Thurman, RG TI Increased susceptibility to alcoholic hepatitis in female rats is associated with elevated free radical formation. SO HEPATOLOGY LA English DT Meeting Abstract C1 UNIV N CAROLINA,DEPT PHARMACOL,HEPATOBIOL & TOXICOL LAB,CHAPEL HILL,NC 27599. NIEHS,RES TRIANGLE PK,NC 27709. NR 0 TC 1 Z9 1 U1 1 U2 1 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 1249 EP 1249 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28501247 ER PT J AU Atchison, CR Hargus, SJ Aronson, JF Hoffmann, WE Daiker, DH Pohl, LR West, AB Moslen, MT AF Atchison, CR Hargus, SJ Aronson, JF Hoffmann, WE Daiker, DH Pohl, LR West, AB Moslen, MT TI Diclofenac decreases serum alkaline phosphatase isozymes and forms protein adducts in multiple tissues. SO HEPATOLOGY LA English DT Meeting Abstract C1 UTMB,DEPT PATHOL,GALVESTON,TX. NIH,LAB MOL IMMUNOL,BETHESDA,MD 20892. COLL VET MED,DEPT PATHOBIOL,URBANA,IL. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 1361 EP 1361 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28501359 ER PT J AU Bezerra, JA Bove, K Wang, MH Degen, SJF AF Bezerra, JA Bove, K Wang, MH Degen, SJF TI Hepatocyte growth factor-like protein is expressed preferentially in pericentral hepatocytes and decreases during liver regeneration and auto-immune hepatitis. SO HEPATOLOGY LA English DT Meeting Abstract C1 UNIV CINCINNATI,CHILDRENS HOSP RES FDN,CINCINNATI,OH. UNIV CINCINNATI,DEPT PATHOL,CINCINNATI,OH. NCI,FREDERICK,MD 21701. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 1430 EP 1430 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28501429 ER PT J AU Bonkovsky, H PohFitzpatrick, M Tattrie, C DiBisceglie, A AF Bonkovsky, H PohFitzpatrick, M Tattrie, C DiBisceglie, A TI Porphyria cutanea tarda and hepatitis C in the USA. SO HEPATOLOGY LA English DT Meeting Abstract C1 UNIV MASSACHUSETTS,MED CTR,WORCESTER,MA. COLUMBIA UNIV,NEW YORK,NY. NIH,BETHESDA,MD 20892. NR 0 TC 1 Z9 1 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 1440 EP 1440 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28501438 ER PT J AU Peterson, TC FernandezSalguero, P Gonzalez, FJ AF Peterson, TC FernandezSalguero, P Gonzalez, FJ TI Hepatic fibrosis as assessed by collagen deposition in AHR knockout mice compared to three animal models of fibrosis. SO HEPATOLOGY LA English DT Meeting Abstract C1 DALHOUSIE UNIV,DEPT MED,HALIFAX,NS,CANADA. DALHOUSIE UNIV,DEPT PHARMACOL,HALIFAX,NS B3H 4H7,CANADA. NCI,MOL CARCINOGENESIS LAB,NIH,BETHESDA,MD 20892. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 1740 EP 1740 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28501735 ER PT J AU Ping, A Sun, SH Winkler, C OBrien, SJ AF Ping, A Sun, SH Winkler, C OBrien, SJ TI Mutations in the core promoter and precore regions of HBV in the hepatocellular carcinoma and liver cirrhosis tissues. SO HEPATOLOGY LA English DT Meeting Abstract C1 LANZHOU MED COLL,LANZHOU,PEOPLES R CHINA. NCI,VIRAL CARCINOGENESIS LAB,FREDERICK,MD 21701. 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 SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD OCT PY 1996 VL 24 IS 4 SU S BP 1744 EP 1744 PN 2 PG 1 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VL285 UT WOS:A1996VL28501739 ER PT J AU Stone, D Ning, Y Guan, XY KaiserKupfer, M WynshawBoris, A Biesecker, L AF Stone, D Ning, Y Guan, XY KaiserKupfer, M WynshawBoris, A Biesecker, L TI Characterization of familial partial 10p trisomy by chromosomal microdissection, FISH, and microsatellite dosage analysis SO HUMAN GENETICS LA English DT Article ID POLYMERASE CHAIN-REACTION; INSITU HYBRIDIZATION; MARKER CHROMOSOMES; RAPID GENERATION; ORIGIN; GENE; TRANSLOCATIONS; IDENTIFICATION; PROBES; PCR AB Unbalanced translocations are a frequent cause of multiple congenital anomalies in children. Translocations as small as 2-5 Mb of DNA are detectable by G-banding under optimal conditions. Some of these small translocations are visible but cannot be characterized cytogenetically due to the lack of characteristic banding on Giemsa preparations. We have combined chromosomal microdissection and fluorescence in situ hybridization (FISH) to identify the origin of a small translocated segment in three members of a family with a derivative chromosome 9 and multiple anomalies, including several ophthalmologic anomalies. We microdissected the abnormal region of the derivative 9 chromosome and used this DNA to generate a FISH probe. This probe hybridized to distal 10p on the metaphase spread of the proband, indicating the origin of the translocated segment. A whole 10p FISH probe confirmed the origin by hybridizing to the translocated segment of the derivative chromosome. FISH was then performed with a whole chromosome 9 painting probe and excluded the presence of a reciprocal, balancing translocation. We then studied the chromosome 10 partial duplication with microsatellite markers to better characterize the chromosomal segment that caused these phenotypic features, By examining the involved areas with distal 10p and 9p microsatellite markers, we were able to demonstrate a minimum of 9 Mb of trisomic 10p DNA with a chromosomal breakpoint between 10p14-10p15. We then compared this family's clinical findings to those of individuals with partial 10p trisomy who had been reported in the literature. The clinical phenotypes seen in this family are similar to, but milder than, the phenotypes of persons with the larger partial trisomies of 10p that were diagnosable by cytogenetic analysis alone, This study shows that microdissection and DNA markers can be used to precisely define small translocations that are difficult to identify by conventional G-banded chromosome analysis. C1 NCHGR,LAB GENET DIS RES,NIH,BETHESDA,MD 20892. NCHGR,DIAGNOST DEV BRANCH,NIH,BETHESDA,MD 20892. NCHGR,CANC GENET LAB,NIH,BETHESDA,MD 20892. NEI,NIH,BETHESDA,MD 20892. RI Guan, Xin-Yuan/A-3639-2009 OI Guan, Xin-Yuan/0000-0002-4485-6017 NR 21 TC 17 Z9 17 U1 0 U2 0 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 SN 0340-6717 J9 HUM GENET JI Hum. Genet. PD OCT PY 1996 VL 98 IS 4 BP 396 EP 402 DI 10.1007/s004390050228 PG 7 WC Genetics & Heredity SC Genetics & Heredity GA VD829 UT WOS:A1996VD82900002 PM 8792811 ER PT J AU Carrington, M Wade, J AF Carrington, M Wade, J TI Selection of transplant donors based on MHC microsatellite data SO HUMAN IMMUNOLOGY LA English DT Editorial Material ID CLASS-II REGION; H-Y; SEQUENCE; ANTIGEN; GENES; DNA; IDENTIFICATION; RECOMBINATION; D6S105; MAP C1 UNIV TORONTO,FAC MED,TTH REG HISTOCOMPATIBIL LAB,TORONTO,ON,CANADA. RP Carrington, M (reprint author), NCI,FREDERICK CANC RES & DEV CTR,SAIC FREDERICK,BIOL CARCINOGENESIS & DEV PROGRAM,POB B,FREDERICK,MD 21702, USA. NR 20 TC 8 Z9 8 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 SN 0198-8859 J9 HUM IMMUNOL JI Hum. Immunol. PD OCT PY 1996 VL 50 IS 2 BP 151 EP 154 DI 10.1016/0198-8859(96)00118-8 PG 4 WC Immunology SC Immunology GA VL239 UT WOS:A1996VL23900010 PM 8891740 ER PT J AU Allikmets, R Gerrard, B Hutchinson, A Dean, M AF Allikmets, R Gerrard, B Hutchinson, A Dean, M TI Characterization of the human ABC superfamily: Isolation and mapping of 21 new genes using the expressed sequence tags database SO HUMAN MOLECULAR GENETICS LA English DT Article ID MULTIDRUG-RESISTANCE; CYSTIC-FIBROSIS; TRANSPORTERS; CLONING; PEROXISOME; SUBUNITS; GENETICS; PROGRAMS; COMPLEX; REGION AB As an approach to characterizing all human ATP-binding cassette (ABC) superfamily genes, a search of the human expressed sequence tag (EST) database was performed using sequences from known ABC genes, A total of 105 clones, containing sequences of potential ABC genes, were identified, representing 21 distinct genes, This brings the total number of characterized human ABC genes from 12 to 33, The new ABC genes were mapped by PCR on somatic cell and radiation hybrid panels and yeast artificial chromosomes (YACs), The genes are located on human chromosomes 1, 2, 3, 4, 6, 7, 10, 12, 13, 14, 16, 17 and X; at locations distinct from previously mapped members of the superfamily. The characterized genes display extensive diversity in sequence and expression pattern and this information was utilized to determine potential structural, functional and evolutionary relationships to previously characterized members of the ABC superfamily. C1 NCI, VIRAL CARCINOGENESIS LAB, SAIC FREDERICK, FREDERICK CANC RES & DEV CTR, FREDERICK, MD 21702 USA. NCI, INTRAMURAL RES SUPPORT PROGRAM, SAIC FREDERICK, FREDERICK CANC RES & DEV CTR, FREDERICK, MD 21702 USA. RI Dean, Michael/G-8172-2012 OI Dean, Michael/0000-0003-2234-0631 NR 39 TC 232 Z9 241 U1 0 U2 3 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0964-6906 EI 1460-2083 J9 HUM MOL GENET JI Hum. Mol. Genet. PD OCT PY 1996 VL 5 IS 10 BP 1649 EP 1655 DI 10.1093/hmg/5.10.1649 PG 7 WC Biochemistry & Molecular Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Genetics & Heredity GA VL702 UT WOS:A1996VL70200018 PM 8894702 ER PT J AU Wayne, S DerKaloustian, VM Schloss, M Polomeno, R Scott, DA Hejtmancik, JF Sheffield, VC Smith, RJH AF Wayne, S DerKaloustian, VM Schloss, M Polomeno, R Scott, DA Hejtmancik, JF Sheffield, VC Smith, RJH TI Localization of the Usher syndrome type ID gene (Ush1D) to chromosome 10 SO HUMAN MOLECULAR GENETICS LA English DT Article ID RECESSIVE TRAITS; ASSIGNMENT; DEAFNESS; LINKAGE; COLLAGEN; MUTATION; MAPS; DNA AB The Usher syndromes (USH) are a group of autosomal recessive diseases characterized by progressive pigmentary retinopathy and sensorineural hearing loss, Five USH genes have been mapped and at least one additional gene is known to exist, By homozygosity mapping in a consanguineous family, a sixth USH gene has been localized, Clinical findings in the four affected children are consistent with established diagnostic criteria for Ush1, Linkage to known USH loci was excluded, and using two genomic DNA pools, one from the affected children and the other from the parents, 161 polymorphic markers evenly spaced across the autosomal human genome were screened, The location of the Ush1 D gene was defined by the only region showing homozygosity by descent in the affected siblings, a15 cM interval on chromosome 10q bounded by D10S529 and D10S573. C1 UNIV IOWA,DEPT OTOLARYNGOL,IOWA CITY,IA 52242. UNIV IOWA,DEPT PEDIAT,IOWA CITY,IA 52242. MCGILL UNIV,DEPT PEDIAT,MONTREAL,PQ H3A 2T5,CANADA. MCGILL UNIV,DEPT HUMAN GENET,MONTREAL,PQ H3A 2T5,CANADA. MCGILL UNIV,DEPT OTOLARYNGOL,MONTREAL,PQ H3A 2T5,CANADA. MCGILL UNIV,DEPT OPHTHALMOL,MONTREAL,PQ H3A 2T5,CANADA. NEI,BETHESDA,MD 20892. FU NIDCD NIH HHS [1RO1 DC02046-01] NR 25 TC 53 Z9 53 U1 1 U2 3 PU OXFORD UNIV PRESS UNITED KINGDOM PI OXFORD PA WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP SN 0964-6906 J9 HUM MOL GENET JI Hum. Mol. Genet. PD OCT PY 1996 VL 5 IS 10 BP 1689 EP 1692 DI 10.1093/hmg/5.10.1689 PG 4 WC Biochemistry & Molecular Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Genetics & Heredity GA VL702 UT WOS:A1996VL70200025 PM 8894709 ER PT J AU Myers, RB Srivastava, S Oelschlager, DK Brown, D Grizzle, WE AF Myers, RB Srivastava, S Oelschlager, DK Brown, D Grizzle, WE TI Expression of nm23-H1 in prostatic intraepithelial neoplasia and adenocarcinoma SO HUMAN PATHOLOGY LA English DT Article DE prostatic intraepithelial neoplasia; prostatic adenocarcinoma; nm23-H1 ID HUMAN BREAST-CANCER; REDUCED EXPRESSION; MESSENGER-RNA; CARCINOMA; GENE; METASTASIS; ASSOCIATION; DYSPLASIA; FREQUENCY; CELLS AB The product of the nm23-H1 gene has been reported to be related to the metastatic potential of several tumors. Although several studies have characterized the expression of the nm23-H1 gene product in prostatic adenocarcinoma, little is known of the expression of this protein in prostatic intraepithelial neoplasia (PIN), a putative precancerous lesion, The authors used immunohistochemistry to examine the expression of the nm23-H1 protein in PIN as well as benign and malignant prostatic tissue. A monoclonal antibody to nm23-H1 (Novocastra, Newcastle upon Tyne, UK, clone 37.6) and a biotin/strepavidin detection system were used for antigen localization. Weak to moderate immunostaining was consistently detected in the benign glandular epithelium of 28 radical prostatectomy specimens. In contrast, strong immunostaining was detected in the glandular epithelium in PIN lesions of 19 radical prostatectomy specimens examined, Strong immunostaining was also frequently detected in the malignant cells of 39 localized prostatic adenocarcinomas, as well as 7 metastatic lesions. These findings show a phenotypic similarity of PIN to prostatic adenocarcinoma with respect to nm23-H1 expression. Furthermore, strong expression of nm23-H1 likely represents an early event in the development of prostatic adenocarcinoma. Copyright (C) 1996 by W.B. Saunders Company C1 UNIV ALABAMA,DEPT PATHOL,BIRMINGHAM,AL 35294. UNIV ALABAMA,DEPT EPIDEMIOL,BIRMINGHAM,AL 35294. NCI,BETHESDA,MD 20892. FU NCI NIH HHS [N01-CN-15340-02] NR 34 TC 19 Z9 19 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0046-8177 J9 HUM PATHOL JI Hum. Pathol. PD OCT PY 1996 VL 27 IS 10 BP 1021 EP 1024 DI 10.1016/S0046-8177(96)90278-6 PG 4 WC Pathology SC Pathology GA VN304 UT WOS:A1996VN30400006 PM 8892585 ER PT J AU Nanagara, R Duray, PH Schumacher, HR AF Nanagara, R Duray, PH Schumacher, HR TI Ultrastructural demonstration of spirochetal antigens in synovial fluid and synovial membrane in chronic Lyme disease: Possible factors contributing to persistence of organisms SO HUMAN PATHOLOGY LA English DT Article DE Lyme disease; synovial; ultrastructure ID BORRELIA-BURGDORFERI ANTIGENS; POLYMERASE CHAIN-REACTION; PROTECTIVE IMMUNITY; JOINT FLUID; ARTHRITIS; MICE; INTERLEUKIN-1; BLOOD; FIBROBLASTS; MECHANISMS AB To perform the first systematic electronmicroscopic (EM) and immunoelectron microscopy (DEM) study of the pathological changes and the evidence of spirochete presence in synovial membranes and synovial fluid (SF) cells of patients with chronic Lyme arthritis. EM examination was performed on four synovial membrane and eight SF cell samples from eight patients with chronic Lyme disease. Spirochetal antigens in the samples were sought by DEM using monoclonal antibody to Borrelia burgdorferi outer surface protein A (OspA) as the immunoprobe, Prominent ultrastructural findings were surface fibrin-like material, thickened synovial lining cell layer and signs of vascular injury. Borrelia-like structures were identified in all four synovial membranes and in two of eight SF cell samples. The presence of spirochetal antigens was confirmed by IEM in all four samples studied (one synovial membrane and three SF cell samples), OspA labelling was in perivascular areas, deep synovial stroma among collagen bundles, and in vacuoles of fibroblasts in synovial membranes; and in cytophagosomes of mononuclear cells in SF cell samples. Electron microscopy adds further evidence for persistence of spirochetal antip ens in the joint in chronic Lyme disease, Locations of spirochetes or spirochetal antigens both intracellulary and extracellulary in deep synovial connective tissue as reported here suggest sites at which spirochaetes may elude host immune response and antibiotic treatment. Copyright (C) 1996 by W.B. Saunders Company C1 VET AFFAIRS MED CTR,RHEUMATOL IMMUNOL CTR,ARTHRIT RES 151K,PHILADELPHIA,PA 19104. KHON KAEN UNIV,FAC MED,DEPT MED,ALLERGY IMMUNOL RHEUMATOL DIV,KHON KAEN,THAILAND. UNIV PENN,SCH MED,DEPT MED,PHILADELPHIA,PA 19104. NCI,PATHOL LABS,NIH,BETHESDA,MD. NR 57 TC 20 Z9 20 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0046-8177 J9 HUM PATHOL JI Hum. Pathol. PD OCT PY 1996 VL 27 IS 10 BP 1025 EP 1034 DI 10.1016/S0046-8177(96)90279-8 PG 10 WC Pathology SC Pathology GA VN304 UT WOS:A1996VN30400007 PM 8892586 ER PT J AU Hughes, AL Yeager, M Carrington, M AF Hughes, AL Yeager, M Carrington, M TI Peptide binding function and the paradox of HLA disease associations SO IMMUNOLOGY AND CELL BIOLOGY LA English DT Article DE HLA/disease associations; major histocompatibility complex; peptide binding; recombination ID DEPENDENT DIABETES-MELLITUS; CLASS-I LOCI; NUCLEOTIDE SUBSTITUTION; 3-DIMENSIONAL STRUCTURE; OVERDOMINANT SELECTION; RHEUMATOID-ARTHRITIS; MHC; SUSCEPTIBILITY; ANTIGENS; ALLELES AB Many studies have found associations between HLA loci and susceptibility or resistance to both infectious and autoimmune diseases, but often subsequent studies fail to find the same association. Such inconsistencies are-not surprising if we consider what is known of the population biology and evolution of HLA genes, especially the consequences of natural selection favouring heterozygosity in the peptide binding regions (PER) of HLA molecules. Because of past recombination events and/or convergent evolution, alleles that are not closely related in overall sequence map come to resemble each other in the PBR. Because peptide binding is likely to be important for the role of HLA molecules in both infectious and autoimmune disease, a strategy of searching for HLA disease associations that groups alleles in functional categories based on PER motifs may prove more successful than conventional strategics, Likewise, in developing approaches for molecular typing, it may be advisable to develop methods that group alleles on the basis of shared PER motifs rather than simply on the basis of shared primer sites in less functionally important regions. C1 PENN STATE UNIV,INST MOL EVOLUT GENET,UNIVERSITY PK,PA 16802. NCI,BIOL CARCINOGENESIS & DEV PROGRAM,SAIC FREDERICK,FREDERICK CANC RES & DEV CTR,FREDERICK,MD 21701. RP Hughes, AL (reprint author), PENN STATE UNIV,DEPT BIOL,MUELLER LAB,UNIVERSITY PK,PA 16802, USA. FU NIGMS NIH HHS [K04-GM00614, RD1-GM43940] NR 31 TC 28 Z9 29 U1 1 U2 2 PU BLACKWELL SCIENCE 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 OCT PY 1996 VL 74 IS 5 BP 444 EP 448 DI 10.1038/icb.1996.74 PG 5 WC Cell Biology; Immunology SC Cell Biology; Immunology GA VQ232 UT WOS:A1996VQ23200008 PM 8912007 ER PT J AU Gu, XX Tsai, CM Ueyama, T Barenkamp, SJ Robbins, JB Lim, DJ AF Gu, XX Tsai, CM Ueyama, T Barenkamp, SJ Robbins, JB Lim, DJ TI Synthesis, characterization, and immunologic properties of detoxified lipooligosaccharide from nontypeable Haemophilus influenzae conjugated to proteins SO INFECTION AND IMMUNITY LA English DT Article ID NONTYPABLE HEMOPHILUS-INFLUENZAE; OUTER-MEMBRANE PROTEIN; ACUTE OTITIS-MEDIA; BACTERICIDAL ANTIBODY; BIOLOGIC ACTIVITY; SERUM; LIPOPOLYSACCHARIDES; PROTECTION; CHILDREN; IMMUNIZATION AB Nontypeable Haemophilus influenzae (NTHi) is an important cause of otitis media in children and of pneumonitis in adults with depressed resistance. Lipooligosaccharide (LOS) is a major surface antigen of NTHi and elicits bactericidal and opsonic antibodies, We prepared detoxified LOS (dLOS) protein conjugates from NTHi for use as experimental vaccines. LOS from NTHi 9274 was treated with anhydrous hydrazine and had its toxicity reduced to clinically acceptable levels, dLOS was bound to tetanus toroid (TT) or high-molecular-weight proteins (HMPs) from NTHi through a linker of adipic acid dihydrazide to form dLOS-TT or dLOS-HMP. The molar ratio of the dLOS to protein carriers ranged from 26:1 to 50:1. The antigenicity of the conjugates was similar to that of the LOS alone as determined by double immunodiffusion. Subcutaneous or intramuscular injection of the conjugates elicited a 28- to 486-fold rise in the level of immunoglobulin G antibodies in mice to the homologous LOS after two or three injections and a 169- to 243-fold rise in the level of immunoglobulin G antibodies in rabbits after two injections. The immunogenicity of the conjugates in mice and rabbits was enhanced by formulation with monophosphoryl lipid A plus trehalose dimycolate. In rabbits, conjugate-induced LOS antibodies induced complement-mediated bactericidal activity against the homologous strain 9274 and prototype strain 3189, These results indicate that a detoxified LOS-protein conjugate is a candidate vaccine for otitis media and pneumonitis caused by NTHi. C1 US FDA,CTR BIOL EVALUAT & RES,BETHESDA,MD 20892. NICHHD,BETHESDA,MD 20892. ST LOUIS UNIV,SCH MED,DEPT PEDIAT,ST LOUIS,MO 63104. ST LOUIS UNIV,CARDINAL GLENNON CHILDRENS HOSP,PEDIAT RES INST,ST LOUIS,MO 63104. RP Gu, XX (reprint author), NIDOCD,NIH,VACCINE DEV UNIT,LAB CELLULAR BIOL,5 RES CT,RM 2A31,ROCKVILLE,MD 20850, USA. OI Barenkamp, Stephen/0000-0002-3054-8910 NR 54 TC 43 Z9 44 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD OCT PY 1996 VL 64 IS 10 BP 4047 EP 4053 PG 7 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA VJ794 UT WOS:A1996VJ79400014 PM 8926067 ER PT J AU Cohen, D Ashkenazi, S Green, M Lerman, Y Slepon, R Robin, G Orr, N Taylor, DN Sadoff, JC Chu, CY Shiloach, J Schneerson, R Robbins, JB AF Cohen, D Ashkenazi, S Green, M Lerman, Y Slepon, R Robin, G Orr, N Taylor, DN Sadoff, JC Chu, CY Shiloach, J Schneerson, R Robbins, JB TI Safety and immunogenicity of investigational Shigella conjugate vaccines in Israeli volunteers SO INFECTION AND IMMUNITY LA English DT Article ID LIPOPOLYSACCHARIDE O-ANTIGEN; INFLUENZAE TYPE-B; SERUM ANTIBODIES; CAPSULAR POLYSACCHARIDE; IMMUNOGLOBULIN-A; PERIPHERAL-BLOOD; SONNEI; EPIDEMIOLOGY; TRANSMISSION; IMMUNIZATION AB The safety and immunogenicity of investigational conjugates, composed of the O-specific polysaccharides of Shigella sonnei and Shigella flexneri type 2a covalently bound to Pseudomonas aeruginosa recombinant exoprotein A (rEPA), were evaluated in 192 Israeli soldiers. None had significant local reactions or fever, Fourteen days after injection, 90% of S. sonnei-rEPA recipients and 73 to 77% of S. flexneri-rEPA recipients had a fourfold or greater increase in serum immunoglobulin G (IgG) and IgA anti-lipopolysaccharide (anti-LPS) levels; at 2 years, these remained higher than at prevaccination (P < 0.01), There was a fourfold or greater increase in IgM anti-LPS in 20% of vaccinees at 2 weeks, but levels returned to prevaccination values at 6 to 12 months, IgG was the highest and most sustained class of LPS antibodies. Reinjection at day 42 did not boost antibody levels, Eighteen of 23 (78%) who received S. sonnei-rEPA and 13 of 19 (68%) who received S. flexneri-rEPA had significant IgA-secreting cell responses, Significant IgG antibody-secreting cell responses were detected in 19 of 23 (83%) and 11 of 19 (58%) volunteers following vaccination with S. sonnei-rEPA and S. flexneri 2a-rEPA, respectively, On the basis of these data, further evaluation of the Shigella conjugates for protective efficacy in field trials in Israel was started. C1 TEL AVIV UNIV,SACKLER SCH MED,IL-69978 RAMAT AVIV,TEL AVIV,ISRAEL. WALTER REED ARMY MED CTR,WALTER REED ARMY INST RES,WASHINGTON,DC 20307. NICHHD,DEV & MOL IMMUN LAB,BETHESDA,MD 20892. NIDDK,BIOTECHNOL UNIT,BETHESDA,MD 20892. RP Cohen, D (reprint author), ISRAEL DEF FORCES MED CORPS,MIL POST 02149,JERUSALEM,ISRAEL. NR 25 TC 47 Z9 53 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD OCT PY 1996 VL 64 IS 10 BP 4074 EP 4077 PG 4 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA VJ794 UT WOS:A1996VJ79400018 PM 8926071 ER PT J AU Whittaker, CJ Clemans, DL Kolenbrander, PE AF Whittaker, CJ Clemans, DL Kolenbrander, PE TI Insertional inactivation of an intrageneric coaggregation-relevant adhesin locus from Streptococcus gordonii DL1 (Challis) SO INFECTION AND IMMUNITY LA English DT Article ID ACTINOMYCES-NAESLUNDII PK606; CELL-SURFACE POLYPEPTIDE; BINDING PROTEIN SSAB; NUCLEOTIDE-SEQUENCE; ORAL STREPTOCOCCI; SALIVARY-AGGLUTININ; DNA FRAGMENTS; SANGUIS 12; GENE; CLONING AB Transposon Tn916 was used to insertionally inactivate a coaggregation-relevant locus of Streptococcus gordonii DL1 (Challis). One mutant (F11) was isolated that lost the ability to coaggregate with the streptococcal partners of DL1 but retained the ability to coaggregate with partners belonging to other genera, A probe specific for the region flanking the Tn916 insertion was used to isolate a locus-specific fragment from a chromosomal lambda library, Southern analysis of the resulting phagemids revealed that a 0.5-kb EcoRI fragment hybridized with the F11 probe. Cloning of the 0.5-kb EcoRI fragment into the E. coli-streptococcal insertion vector p Omega yielded pCW4, which was used to insertionally inactivate the putative coaggregation-relevant gene in DL1. Insertion mutants showed altered coaggregation with streptococci but retained wild-type coaggregation properties with other genera of bacteria. Comparison of immunoblots of cell surface proteins showed a 100-kDa protein in DL1 which was not detected in the Tn916 and pCW4 insertion mutants. These results indicate that the 0.5-kb EcoRI fragment is part of an adhesin-relevant locus that is involved in the production of a 100-kDa protein at the cell surface. C1 NIDR,MICROBIAL ECOL LAB,NIH,BETHESDA,MD 20892. NR 49 TC 22 Z9 24 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD OCT PY 1996 VL 64 IS 10 BP 4137 EP 4142 PG 6 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA VJ794 UT WOS:A1996VJ79400027 PM 8926080 ER PT J AU Nagineni, CN Pardhasaradhi, K Martins, MC Detrick, B Hooks, JJ AF Nagineni, CN Pardhasaradhi, K Martins, MC Detrick, B Hooks, JJ TI Mechanisms of interferon-induced inhibition of Toxoplasma gondii replication in human retinal pigment epithelial cells SO INFECTION AND IMMUNITY LA English DT Article ID GAMMA-INTERFERON; OCULAR TOXOPLASMOSIS; HUMAN-FIBROBLASTS; HUMAN MACROPHAGES; BETA-INTERFERON; GENE-EXPRESSION; PROTECTIVE ROLE; IFN-GAMMA; GROWTH; CYTOKINES AB Inflammation associated with retinochoroiditis is a major complication of ocular toxoplasmosis in infants and immunocompetent individuals. Moreover, Toxoplasma gondii-induced retinal disease causes serious complications in patients with AIDS and transplant patients. The retinal pigment epithelial (RPE) cell is an important regulatory cell within the retina and is one of the cells infected with T. gondii in in vivo. We have developed a human RPE (HRPE) cell in vitro model system to evaluate T. gondii replication and the regulation of this replication by cytokines. T. gondii replication was quantitated by counting the foci of infection (plaque formation) and the numbers of tachyzoites released into the supernatant fluids. Pretreatment of cultures with recombinant human tumor necrosis factor alpha, alpha interferon (IFN-alpha), IFN-beta, or IFN-gamma for 24 h prior to inoculation inhibited T. gondii replication in a dose-dependent manner. Of these cytokines, IFN-gamma was the most potent, and T. gondii replication was completely inhibited at a concentration of 100 U/ml. The anti-toxoplasmotic activity of IFN-gamma was significantly blocked by monoclonal antibody to IFN-gamma. Treatment of the cultures with IFN-gamma from day 1 or 2 postinoculation with T. gondii also offered protection against the parasite. The anti toxoplasmotic activity of tumor necrosis factor alpha or IFN-alpha, -beta, or -gamma in these cultures was found to be independent of the nitric oxide (NO) pathway, since NO production was not found in HRPE cells treated with these cytokines. However, addition of tryptophan to IFN-gamma-treated cells significantly reversed the inhibitory effects of IFN-gamma, suggesting that IFN-gamma acts by depleting cellular tryptophan. This effect was further confirmed by reverse transcription-PCR and Northern (RNA) blot analysis, which indicated induction of indoleamine 2,3-dioxygenase (IDO), an enzyme that converts tryptophan to kynurenine. These results indicated that interferons inhibited T. gondii replication in HRPE by NO-independent but IDO-dependent mechanisms. This in vitro model of T. gondii replication in HRPE may be useful in evaluating the effects of cytokines and drugs on T. gondii replication within the retina. C1 NEI,IMMUNOL LAB,IMMUNOL & VIROL SECT,NIH,BETHESDA,MD 20892. WALTER REED ARMY MED CTR,DIV BIOCHEM,WASHINGTON,DC 20307. GEORGE WASHINGTON UNIV,MED CTR,DEPT PATHOL,WASHINGTON,DC 20037. NR 54 TC 103 Z9 108 U1 1 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD OCT PY 1996 VL 64 IS 10 BP 4188 EP 4196 PG 9 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA VJ794 UT WOS:A1996VJ79400034 PM 8926087 ER PT J AU Rockey, DD Fischer, ER Hackstadt, T AF Rockey, DD Fischer, ER Hackstadt, T TI Temporal analysis of the developing Chlamydia psittaci inclusion by use of fluorescence and electron microscopy SO INFECTION AND IMMUNITY LA English DT Article ID INFECTED-CELLS; HOST-CELLS; TRACHOMATIS; MEMBRANE; PROTEIN; SPHINGOLIPIDS; FUSION AB The chlamydiae are obligate intracellular parasites that develop and multiply within a vacuole (termed an inclusion) that does not fuse with lysosomes, Inclusion morphology varies dramatically among the different chlamydiae, particularly within the species Chlamydia psittaci. Some strains develop within a single vacuole, while the mature inclusion of other strains consists of several distinct lobes, each filled with chlamydial developmental forms, The development of this lobed structure was investigated in HeLa cells infected with the guinea pig inclusion conjunctivitis (GPIC) strain of C. psittaci, We employed two recently described probes for the chlamydial inclusion to study the development of these unique lobed structures, The novel probes were an antiserum directed at a protein localized to the GPIC inclusion membrane (anti-IncA) and the fluorescent sphingolipid {N-[7-(4-nitrobenzo-2-oxa-1,3-)]} aminocaproyl sphingosine (NBD-ceramide). Lobed inclusions developed in cells infected at very low multiplicities of infection, suggesting that the structure is not a function of infection by more than one elementary body (EB), Double-label fluorescent-antibody analysis with anti-IncA and an antibody directed at a chlamydial outer membrane protein showed that, prior to 18 h postinfection (p.i.), the inclusion membrane and the chlamydial membrane were tightly associated, After 18 to 20 h p.i., the lobes began to expand and fill with developmental forms and the inclusion membrane and chlamydial membrane became distinct. At times from 8 to 48 h p.i., GPIC inclusions were shown to receive fluorescent derivatives of NBD-ceramide and to he localized to the perinuclear region of the host cell. Labeled lectins with affinity for carbohydrate moieties localized to the Golgi apparatus showed that the lobes of mature inclusions surround the Golgi apparatus, Labeling with NBD-ceramide and the Golgi apparatus-specific lectins therefore demonstrated a functional and physical association of the inclusion with the Golgi apparatus throughout the developmental cycle, Collectively, these results lead to a model for the development of the lobed chlamydial inclusion, We propose that the lobed structure is a result of division of inclusions occurring in parallel with the multiplication of reticulate bodies (RE) early in the developmental cycle, The division of inclusions slows or stops in mid-cycle, and dividing RE accumulate within the enlarging lobes, The RE then differentiate to EBs, the inclusion and cell are lysed, and EBs are freed to infect another cell. RP Rockey, DD (reprint author), NIAID,ROCKY MT LABS,HAMILTON,MT 59840, USA. NR 25 TC 53 Z9 54 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD OCT PY 1996 VL 64 IS 10 BP 4269 EP 4278 PG 10 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA VJ794 UT WOS:A1996VJ79400046 PM 8926099 ER PT J AU Sugimura, Y Foster, BA Hom, YK Lipschutz, JH Rubin, JS Finch, PW Aaronson, SA Hayashi, N Kawamura, J Cunha, GR AF Sugimura, Y Foster, BA Hom, YK Lipschutz, JH Rubin, JS Finch, PW Aaronson, SA Hayashi, N Kawamura, J Cunha, GR TI Keratinocyte growth factor (KGF) can replace testosterone in the ductal branching morphogenesis of the rat ventral prostate SO INTERNATIONAL JOURNAL OF DEVELOPMENTAL BIOLOGY LA English DT Article DE keratinocyte growth factor; mesenchymal-epithelial; interactions; prostate; testosterone; branching morphogenesis; epithelial growth ID ANDROGEN RECEPTOR EXPRESSION; MESENCHYMAL-EPITHELIAL INTERACTIONS; DNA SYNTHETIC ACTIVITY; FEMINIZED TFM/Y MICE; IN-VIVO; SEMINAL-VESICLE; MOUSE PROSTATE; TESTICULAR FEMINIZATION; POSTNATAL-DEVELOPMENT; ABNORMAL-DEVELOPMENT AB Prostatic growth occurs through ductal elongation and branching into the mesenchyme. Ductal branching morphogenesis in the prostate is elicited by androgens via mesenchymal-epithelial interactions mediated by paracrine influences from mesenchyme. The role of keratinocyte growth factor (KGF) was investigated in the developing prostate as KGF has been suggested to be a paracrine acting factor. KGF transcripts were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) in neonatal rat Ventral prostates (VPs) in vivo, in VPs cultured in vitro, and in isolated VP mesenchyme. KGF receptor was detected in VP's by RT-PCR and was localized specifically to the epithelium by in situ hybridization. KGF was investigated as a potential paracrine mediator during androgen-induced prostatic development by examining neonatal rat VPs cultured for 6 days under serum-free conditions using a basal medium supplemented only with insulin and transferrin. When testosterone (10(-9) to 10(-8) M) was added to the basal medium, VPs grew and underwent ductal branching morphogenesis similar to that in situ. Neutralization of endogenous KGF with a monoclonal antibody to KGF (anti-KGF) or a soluble KGF receptor peptide inhibited androgen-stimulated VP growth (DNA content) and reduced the number of ductal end buds after 6 days of culture. When KGF (50 or 100 ng/ml) was added to the basal medium in the absence of testosterone. VP growth and ductal branching morphogenesis were stimulated. The number of ductal end buds was about 70% of that obtained with an optimal dose of testosterone (10(-8) M), and DNA content of VP's cultured with 100 ng/ml KGF was equivalent to that of glands cultured with testosterone. The stimulatory effect of KGF was partially blocked by cyproterone acetate, a steroidal anti-androgen. These data imply that KGF plays an important role as a mesenchymal paracrine mediator of androgen-induced epithelial growth and ductal branching morphogenesis in the rat VP. C1 UNIV CALIF SAN FRANCISCO, DEPT ANAT, SAN FRANCISCO, CA 94143 USA. AICHI CANC CTR, DEPT UROL, NAGOYA, AICHI 464, JAPAN. NCI, CELLULAR & MOL BIOL LAB, BETHESDA, MD 20892 USA. MT SINAI SCH MED, RUTTENBERG CANC CTR, NEW YORK, NY USA. MIE UNIV, SCH MED, DEPT UROL, TSU, MIE, JAPAN. FU NCI NIH HHS [CA 59831]; NIDDK NIH HHS [DK 45861 +, DK 32157] NR 76 TC 130 Z9 133 U1 0 U2 1 PU U B C PRESS PI BILBAO PA UNIV BASQUE COUNTRY, EDITORIAL SERVICES, PO BOX 1397, E-48080 BILBAO, SPAIN SN 0214-6282 J9 INT J DEV BIOL JI Int. J. Dev. Biol. PD OCT PY 1996 VL 40 IS 5 BP 941 EP 951 PG 11 WC Developmental Biology SC Developmental Biology GA VR175 UT WOS:A1996VR17500003 PM 8946242 ER PT J AU Stoll, J Galdzicki, Z AF Stoll, J Galdzicki, Z TI Reduced expression of voltage-gated sodium channels in neurons cultured from trisomy 16 mouse hippocampus SO INTERNATIONAL JOURNAL OF DEVELOPMENTAL NEUROSCIENCE LA English DT Article DE saxitoxin; sodium channel; trisomy; neuronal cell culture; gene expression ID DORSAL-ROOT GANGLION; CAMP-DEPENDENT PHOSPHORYLATION; RAT-BRAIN; ALPHA-SUBUNIT; MESSENGER-RNAS; SKELETAL-MUSCLE; MODEL; HEART; CELLS AB Voltage-gated sodium channels are responsible for the initial depolarizing phase of the action potential. In hippocampal neurons cultured from trisomy 16 (Ts16) mice (a model for Down's syndrome), the maximum inward conductance mediated by these channels was reduced 47% relative to control diploid neurons. This reduced conductance was reflected in a 35% decrease in binding of radiolabeled saxitoxin, a sodium channel-specific ligand, indicating expression of fewer channels in these neurons. The mRNAs encoding the alpha and beta 1 subunits were, however, present at the same levels in Ts16 neurons and control diploid neurons. Thus, the altered regulation of voltage-gated sodium channels in Ts16 neurons is apparently a post-transcriptional event and possible mechanisms are discussed. C1 NIA,NEUROSCI LAB,BETHESDA,MD 20892. NR 38 TC 9 Z9 9 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, ENGLAND OX5 1GB SN 0736-5748 J9 INT J DEV NEUROSCI JI Int. J. Dev. Neurosci. PD OCT PY 1996 VL 14 IS 6 BP 749 EP 760 DI 10.1016/S0736-5748(96)00051-2 PG 12 WC Developmental Biology; Neurosciences SC Developmental Biology; Neurosciences & Neurology GA VX897 UT WOS:A1996VX89700007 PM 8960982 ER PT J AU Kafadar, K Freedman, LS Goodall, CR Tukey, JW AF Kafadar, K Freedman, LS Goodall, CR Tukey, JW TI Urbanicity-related trends in lung cancer mortality in US counties: White females and white males, 1970-1987 SO INTERNATIONAL JOURNAL OF EPIDEMIOLOGY LA English DT Article DE geographical epidemiology; urbanicity; smoothing; Poisson; exploratory data analysis ID UNITED-STATES; RATES; STATISTICS; PATTERNS; MAPS AB Background. The effect of urbanization on age-adjusted lung cancer mortality rates in US counties is investigated, The data come from National Cancer Institute, and urban trends are estimated in time periods 1970-1979 and 1980-1987, for both white males and white females. To account for possibly different gradients in different parts of the country, the 48 contiguous states are divided into seven regions. Methods. A measure of urbanness, urbanicity, is defined and is used to stratify counties. A multiplicative model is proposed that relates county mortality rates to urbanicity. The residuals from this multiplicative model serve as age- and urban-adjusted rates. Results. Urban-rural gradients are significant for nearly all regions for both white males and white females, diminishing slightly in the latter time period for white males but becoming stronger for white females. Conclusions. The age- and urban-adjusted rates may be used in mapping to investigate geographical patterns that remain after removal of the urban factor. C1 NCI,DIV CANC PREVENT & CONTROL,BIOMETRY BRANCH,BETHESDA,MD 20892. PENN STATE UNIV,DEPT STAT,UNIVERSITY PK,PA 16802. PRINCETON UNIV,DEPT STAT,PRINCETON,NJ 08544. NR 33 TC 3 Z9 3 U1 0 U2 0 PU OXFORD UNIV PRESS UNITED KINGDOM PI OXFORD PA WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP SN 0300-5771 J9 INT J EPIDEMIOL JI Int. J. Epidemiol. PD OCT PY 1996 VL 25 IS 5 BP 918 EP 932 DI 10.1093/ije/25.5.918 PG 15 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA VR165 UT WOS:A1996VR16500003 PM 8921476 ER PT J AU Murphy, EL Wilks, R Hanchard, B Cranston, B Figueroa, JP Gibbs, WN Murphy, JYN Blattner, WA AF Murphy, EL Wilks, R Hanchard, B Cranston, B Figueroa, JP Gibbs, WN Murphy, JYN Blattner, WA TI A case-control study of risk factors for seropositivity to human T-Lymphotropic virus type I (HTLV-I) in Jamaica SO INTERNATIONAL JOURNAL OF EPIDEMIOLOGY LA English DT Article DE HTLV-I infection; risk factors; intrauterine device; sexual transmission; Jamaica ID CELL LEUKEMIA-VIRUS; SEXUAL TRANSMISSION; DETERMINANTS; JAPAN; SEROCONVERSION; SEROPREVALENCE; TRANSFUSION; INFECTION; LYMPHOMA; ANTIBODY AB Background. We investigated behavioural and environmental risk factors for seropositivity to human T-lymphotropic virus type I (HTLV-I). Methods. A nested case-control study of 201 HTLV-I seropositive subjects and 225 age: and sex-matched seronegative controls was performed using questionnaire data from the enrolment visit of a cohort study in 1987-1988. HTLV-I serostatus was confirmed using enzyme-linked immunosorbent assay (ELISA) and Western blot. Results. Among women, the number of lifetime sexual partners (P < 0.05, chi(2) trend) and the number of different men fathering a child by the woman (P < 0.06, chi(2) trend) were associated with HTLV-I seropositivity. Use by the female subject of an intrauterine device (IUD) was associated with an increased risk of seropositivity (odds ratio (OR) = 2.67, 95% confidence interval (CI) : 1.13-6.23); condom use was rare in this population. Among male subjects, a larger number of lifetime sexual partners was also associated with HTLV-I seropositivity (P < 0.05, chi(2) trend). No association was found between HTLV-I seropositivity and educational attainment, income, or occupation. Having been breastfed as a child or receipt of a blood transfusion had elevated but imprecise OR due to very high and low prevalence of the risk factors, respectively. Several variables relating to insect or animal exposure showed no association with HTLV-I seropositivity. Conclusions. These data confirm that heterosexual intercourse is a major route of HTLV-I transmission, but do not support suggestions of insect or environmental vectors. C1 UNIV CALIF SAN FRANCISCO,DEPT MED & EPIDEMIOL & BIOSTAT,SAN FRANCISCO,CA 94143. UNIV W INDIES,TROP METAB RES UNIT,KINGSTON 7,JAMAICA. UNIV W INDIES,DEPT PATHOL,KINGSTON 7,JAMAICA. UNIV W INDIES,DEPT MED,KINGSTON 7,JAMAICA. MINIST HLTH,KINGSTON,JAMAICA. WHO,CH-1211 GENEVA,SWITZERLAND. RES TRIANGLE INST,WASHINGTON,DC. NCI,BETHESDA,MD 20892. RP Murphy, EL (reprint author), UNIV CALIF SAN FRANCISCO,DEPT LAB MED,BOX 0884,SAN FRANCISCO,CA 94143, USA. NR 22 TC 21 Z9 22 U1 0 U2 0 PU OXFORD UNIV PRESS UNITED KINGDOM PI OXFORD PA WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP SN 0300-5771 J9 INT J EPIDEMIOL JI Int. J. Epidemiol. PD OCT PY 1996 VL 25 IS 5 BP 1083 EP 1089 DI 10.1093/ije/25.5.1083 PG 7 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA VR165 UT WOS:A1996VR16500025 PM 8921498 ER PT J AU Murphy, EL Wilks, R Morgan, OS Hanchard, B Cranston, B Figueroa, JP Gibbs, WN Murphy, JYN Blattner, WA AF Murphy, EL Wilks, R Morgan, OS Hanchard, B Cranston, B Figueroa, JP Gibbs, WN Murphy, JYN Blattner, WA TI Health effects of human T-lymphotropic virus type I (HTLV-I) in a Jamaican cohort SO INTERNATIONAL JOURNAL OF EPIDEMIOLOGY LA English DT Article DE HTLV-I infection; HTLV-I associated myelopathy; tropical spastic paraparesis; anaemia; eosinophilia; body mass index ID CELL LEUKEMIA-LYMPHOMA; MYELOPATHY; CARRIERS; INFECTION; RISK; ANTIBODIES; PATIENT; JAPAN; ATL AB Background. Other than adult T-cell leukaemia (ATL) and HTLV-I associated myelopathy (HAM), the health effects of infection with human T-lymphotropic virus type I (HTLV-I) are not well defined. Method A cohort of 201 confirmed HTLV-I seropositive Jamaican food service workers and 225 seronegative controls of similar age and sex from the same population was examined. A health questionnaire, physical examination, and laboratory tests were performed at enrolment into the cohort in 1987-1988. Results. One of 201 HTLV-I seropositives, but no controls were diagnosed with HAM, for a prevalence of 0.5% (95% confidence interval) (CI) 0.01-2.7%); no cases of ATL were diagnosed. While there was no difference in current symptoms, the HTLV-I seropositive group was more likely to report a past medical history of hepatitis or jaundice (OR = 3.49, 95% CI : 0.93-13.08), malaria (OR = 2.13, 95% CI : 0.96-1.73), and dengue fever (OR = 1.37, 95% CI : 0.82-2.29); however, these differences were of borderline statistical significance. Low income HTLV-I seropositive women had lower body weight (P < 0.01) and body mass index (P < 0.009) than their seronegative counterparts; similar differences were seen in the smaller male group. A trend toward higher prevalence of severe anaemia (haemoglobin < 10 g/dl) (12.6% versus 7.7%, P < 0.105) and a significantly lower prevalence of eosinophilia (1.0% versus 6.3%, P < 0.004) was seen among HTLV-I seropositives compared to controls. Conclusions. Although most HTLV-I seropositives are asymptomatic, HAM may be diagnosed in approximately 0.5% of carriers. Chronic HTLV-I infection may also exert subtle effects on body mass and haematological parameters. C1 UNIV CALIF SAN FRANCISCO,DEPT MED,SAN FRANCISCO,CA 94143. UNIV CALIF SAN FRANCISCO,DEPT EPIDEMIOL & BIOSTAT,SAN FRANCISCO,CA 94143. UNIV W INDIES,TROP METAB RES UNIT,KINGSTON 7,JAMAICA. UNIV W INDIES,DEPT PATHOL,KINGSTON 7,JAMAICA. UNIV W INDIES,DEPT MED,KINGSTON 7,JAMAICA. MINIST HLTH,KINGSTON,JAMAICA. RES TRIANGLE INST,WASHINGTON,DC. NCI,BETHESDA,MD 20892. RP Murphy, EL (reprint author), UNIV CALIF SAN FRANCISCO,DEPT LAB MED,BOX 0884,SAN FRANCISCO,CA 94143, USA. NR 34 TC 25 Z9 25 U1 0 U2 0 PU OXFORD UNIV PRESS UNITED KINGDOM PI OXFORD PA WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP SN 0300-5771 J9 INT J EPIDEMIOL JI Int. J. Epidemiol. PD OCT PY 1996 VL 25 IS 5 BP 1090 EP 1097 DI 10.1093/ije/25.5.1090 PG 8 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA VR165 UT WOS:A1996VR16500026 PM 8921499 ER PT J AU Brenner, DJ Hahnfeldt, P Amundson, SA Sachs, RK AF Brenner, DJ Hahnfeldt, P Amundson, SA Sachs, RK TI Interpretation of inverse dose-rate effects for mutagenesis by sparsely ionizing radiation SO INTERNATIONAL JOURNAL OF RADIATION BIOLOGY LA English DT Article ID FISSION-SPECTRUM NEUTRONS; ONCOGENIC TRANSFORMATION; CELL-CYCLE; NEOPLASTIC TRANSFORMATION; L5178Y CELLS; MODEL; INDUCTION; MUTANTS; LETHAL; IRRADIATION AB An inverse dose-rate effect has sometimes been observed for mutagenesis in cells exposed to gamma-rays. We model such data quantitatively with the key assumption that the effect is caused in cycling cells by correlated variations in sensitivity across the cell cycle, for both mutation and killing. We quantify this approach using the LQR (linear-quadratic + resensitization) formalism, which describes the response to radiation of a heterogeneous cell population. This model is applied to an exponentially growing population. We compare its predictions with dose- and dose-rate dependent mutation data and show that it can well fit the observed inverse dose-rate effect, as well as providing an explanation of why inverse dose-rate effects have been seen in some experiments, but not in others. The actual values of the model parameters emerging from the analysis are reasonable in magnitude, based on their biological interpretations. We conclude that the LQR model can quantify cell-cycle redistribution effects without over-parameterization, and that the data favour a correlation explanation of inverse dose-rate effects for mutagenesis by low-LET radiation. It is less clear that this explanation is appropriate to high-LET radiation-induced oncogenic transformation, although all potential explanations of inverse dose-rate effects predict that, at appropriately low doses, no dose-rate effects of any kind are expected. C1 HARVARD UNIV, SCH MED, JOINT CTR RADIAT THERAPY, BOSTON, MA 02115 USA. NATL INST HLTH, MOL PHARMACOL LAB, BETHESDA, MD 20892 USA. UNIV CALIF BERKELEY, DEPT MATH, BERKELEY, CA 94720 USA. UNIV CALIF BERKELEY, DEPT PHYS, BERKELEY, CA 94720 USA. RP Brenner, DJ (reprint author), COLUMBIA UNIV, CTR RADIOL RES, 630 W 168TH ST, NEW YORK, NY 10032 USA. FU NCI NIH HHS [CA-63897, CA-24232] NR 42 TC 25 Z9 26 U1 0 U2 1 PU TAYLOR & FRANCIS LTD PI LONDON PA ONE GUNDPOWDER SQUARE, LONDON, ENGLAND EC4A 3DE SN 0955-3002 J9 INT J RADIAT BIOL JI Int. J. Radiat. Biol. PD OCT PY 1996 VL 70 IS 4 BP 447 EP 458 PG 12 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 VN071 UT WOS:A1996VN07100009 PM 8862456 ER PT J AU Pollack, JD Banzon, J Donelson, K Tully, JG Davis, JW Hackett, KJ Agbanyim, C Miles, RJ AF Pollack, JD Banzon, J Donelson, K Tully, JG Davis, JW Hackett, KJ Agbanyim, C Miles, RJ TI Reduction of benzyl viologen distinguishes genera of the class Mollicutes SO INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY LA English DT Article ID SPIROPLASMA; METABOLISM; OXIDASE AB We tested the ability of 62 growing strains belonging to the class Mollicutes to reduce the redox indicator and free-radical generator 1,1'-dibenzyl-4,4'-bipyridinium dichloride (benzylviologen [BV]) to a blue-violet-purple color, BV was reduced by 12 Acholeplasma species but not by Acholeplasma multiforme PN525(T) (T = type strain), BV was also reduced by five of nine Mesoplasma species and by four of six Entomoplasma species, BV was not reduced by 19 Mycoplasma species, six Spiroplasma species, five unnamed Spiroplasma strains belonging to different serogroups, three Ureaplasma species, and one unnamed Ureaplasma strain, The BV-reducing ability was localized in the membrane of Acholeplasma laidlawii B-PG9 and was dependent on NADH, Reduction of BV could be expressed in mixed cultures, and this activity may be useful for recognizing the contaminating presence of an Acholeplasma species, The reductive BV response may have phylogenetic value, We believe that the test described in this paper readily distinguishes all Acholeplasma species and some Mesoplasma and Entomoplasma species from all Mycoplasma, Spiroplasma, and Ureaplasma species tested. C1 NIAID,MYCOPLASMA SECT,FREDERICK CANC RES FACIL,FREDERICK,MD 21702. CUNY,BRONX COMMUNITY COLL,DEPT BIOL & MED LAB TECHNOL,BRONX,NY 10453. USDA ARS,BELTSVILLE AGR RES CTR,INSECT BIOCONTROL LAB,BELTSVILLE,MD 20705. UNIV LONDON KINGS COLL,DIV LIFE SCI,LONDON W8 7AH,ENGLAND. RP Pollack, JD (reprint author), OHIO STATE UNIV,DEPT MED MICROBIOL & IMMUNOL,333 W 10TH AVE,COLUMBUS,OH 43210, USA. FU NIGMS NIH HHS [SO6-GM08174]; PHS HHS [R01-A133193] NR 24 TC 6 Z9 6 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0020-7713 J9 INT J SYST BACTERIOL JI Int. J. Syst. Bacteriol. PD OCT PY 1996 VL 46 IS 4 BP 881 EP 884 PG 4 WC Microbiology SC Microbiology GA VL867 UT WOS:A1996VL86700007 PM 8863413 ER PT J AU Pollack, JD Williams, MV Banzon, J Jones, MA Harvey, L Tully, JG AF Pollack, JD Williams, MV Banzon, J Jones, MA Harvey, L Tully, JG TI Comparative metabolism of Mesoplasma, Entomoplasma, Mycoplasma, and Acholeplasma SO INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY LA English DT Article ID DEPENDENT NUCLEOSIDE KINASE; TRICARBOXYLIC-ACID CYCLE; MYCOIDES SUBSP MYCOIDES; URACIL-DNA GLYCOSYLASE; GROUP-A STREPTOCOCCI; LAIDLAWII B-PG9; GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE; PHYLOGENETIC ANALYSIS; PURINE METABOLISM; CLASS MOLLICUTES AB Cytoplasmic fractions from species of the Mollicutes genera Entomoplasma, Mesoplasma, Mycoplasma, and Acholeplasma were assayed for NADH oxidase (NADH ox), ATP- and PPi-dependent phosphofructokinase (PFK), ATP- and PPi-dependent deoxyguanosine kinase (dGUOK), thymidine kinase (TK), TMP kinase (TMPK), glucose-6-phosphate dehydrogenase (G6Pde), lactate dehydrogenase (LDH), malate dehydrogenase (MDH), phosphoenolpyruvate carboxylase, hypoxanthine-guanine phosphoribosyl transferase, dUTPase, and uracil-DNA glycosylase (UNG) activities. Membrane fractions were also examined for NADH ox activity. These activities were used as indicators of the presence and relative activities of major Mollicutes metabolic and DNA repair pathways. This was the first study to determine the presence of these enzymes in members of the genera Entomoplasma and Mesoplasma. Using the data obtained, we constructed a preliminary scheme for distinguishing genera of the class Mollicutes on the basis of the results of signature functional enzyme assays. This scheme includes phylogenetic relationships deduced from rRNA analyses, but is more informative with respect to metabolic potential. The criteria used include the presence of PPi-dependent PFK, urease, dUTPase, and dGUOK activities. Entomoplasma ellychniae ELCN-1(T) (T = type strain), Entomoplasma melaleucae M-1(T), Mesoplasma seiffertii F7(T), Mesoplasma entomophilum TAC(T), Mesoplasma florum L1(T), Mycoplasma fermentans PG18(T), and Acholeplasma multilocale PN525(T) were similar in most respects. NADH ox activity was localized in the cytoplasm of these organisms. These strains had ATP-dependent PFK, MDH, LDH, ATP- and PPi-dependent dGUOK, and UNG activities, but not dUTPase or G6Pde activities. In contrast, Acholeplasma equifetale C112(T), Acholeplasma oculi 19L(T), Acholeplasma hippikon C1(T) Acholeplasma modicum PG49(T), and Acholeplasma morum 72-043(T) had membrane-localized NADH ox activity, PPi-dependent PFK, G6Pde, and dUTPase activities, and significantly lower MDH and LDH activities and exhibited a faster rate with PPi than with ATP in the dGUOK reaction. All of the members of the Mollicutes tested had hypoxanthine-guanine phosphoribosyl transferase, phosphoenolpyruvate carboxylase, and (except for Mesoplasma entomophilum TAC(T)) UNG activities. All of the Acholeplasma strains except Acholeplasma multilocale PN525(T) had TK, TMPK, and UNG activities. Mesoplasma entomophilum TAC(T) was distinguished by having no detectable dUTPase, UNG, TK, and TMPK activities, indicating that there is a severe restriction in or an absence of a synthetic route to dTTP. Our data also suggest that A. multilocale PN525(T) is a member of an unrecognized metabolic subgroup of the genus Acholeplasma or is not an Acholeplasma strain. C1 NIAID, MYCOPLASMA SECT, FREDERICK CANC RES FACIL, FREDERICK, MD 21702 USA. RP Pollack, JD (reprint author), OHIO STATE UNIV, DEPT MED MICROBIOL & IMMUNOL, 333 W 10TH AVE, COLUMBUS, OH 43210 USA. FU PHS HHS [R01-A133193] NR 42 TC 20 Z9 20 U1 2 U2 4 PU SOC GENERAL MICROBIOLOGY PI READING PA MARLBOROUGH HOUSE, BASINGSTOKE RD, SPENCERS WOODS, READING RG7 1AG, BERKS, ENGLAND SN 0020-7713 J9 INT J SYST BACTERIOL JI Int. J. Syst. Bacteriol. PD OCT PY 1996 VL 46 IS 4 BP 885 EP 890 PG 6 WC Microbiology SC Microbiology GA VL867 UT WOS:A1996VL86700008 PM 8863414 ER PT J AU Hackett, KJ Whitcomb, RF Clark, TB Henegar, RB Lynn, DE Wagner, AG Tully, JG Gasparich, GE Rose, DL Carle, P Bove, JM Konai, M Clark, EA Adams, JR Williamson, DL AF Hackett, KJ Whitcomb, RF Clark, TB Henegar, RB Lynn, DE Wagner, AG Tully, JG Gasparich, GE Rose, DL Carle, P Bove, JM Konai, M Clark, EA Adams, JR Williamson, DL TI Spiroplasma leptinotarsae sp nov, a mollicute uniquely adapted to its host, the Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae) SO INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY LA English DT Article ID PARASITE RELATIONSHIPS; GENUS SPIROPLASMA; DEFORMATION; DIVERSITY; GROWTH AB Spiroplasma strain LD-1(T) (T = type strain), which was isolated from the gut of a Colorado potato beetle (Leptinotarsa decemlineata) larva collected in Maryland, was serologically distinct from other spiroplasmas. Similar isolates were obtained from other L. decemlineata specimens collected in various parts of North America, in Poland, and in other eastern European countries and from Leptinotarsa texana specimens collected in Texas. Cells of strain LD-1(T), which in early passages were spiral, exhibited exceptionally rapid translational motility. This rapid motility and the spiral shape were lost after extended passage in culture, The organism required serum for growth. Originally isolated in coculture with insect cells in DCCM medium, strain LD-1(T) adapted to several media in the absence of cocultured cells. Use of anaerobic conditions allowed primary isolation in a variety of media. The organism did not grow in serum-free media containing 2% serum fraction, Optimal growth in M1D medium occurred at 30 to 37 degrees C (doubling time, 7.2 h). On solid M1D medium containing 2.0% Noble agar (pH 6.25) at 30 degrees C, strain LD-1(T) produced discrete colonies with numerous satellites. Strain LD-1(T) hydrolyzed arginine, but did not utilize urea; there was evidence of weak fermentation of glucose, The guanine-plus-cytosine content of the DNA was determined to be 25 +/- 1 mol%, and the genome size was 1,085 kb. The results of extensive studies of the ecology of this spiroplasma suggest that it is host specific for Leptinotarsa beetles, Strain LD-1 (= ATCC 43213) is designated the type strain of a new species, Spiroplasma leptinotarsa. C1 USDA ARS,BELTSVILLE AGR RES CTR,INSECT BIOCONTROL LAB,BELTSVILLE,MD 20705. NIAID,MYCOPLASMA SECT,MOL MICROBIOL LAB,FREDERICK CANC RES FACIL,FREDERICK,MD 21702. INSERM,LAB BIOL CELLULAIRE & MOL,F-33883 VILLENAVE DORNON,FRANCE. SUNY STONY BROOK,DEPT ANAT SCI,STONY BROOK,NY 11794. NR 35 TC 17 Z9 18 U1 0 U2 2 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0020-7713 J9 INT J SYST BACTERIOL JI Int. J. Syst. Bacteriol. PD OCT PY 1996 VL 46 IS 4 BP 906 EP 911 PG 6 WC Microbiology SC Microbiology GA VL867 UT WOS:A1996VL86700011 ER PT J AU Hackett, KJ Clark, EA Whitcomb, RF Camp, M Tully, JG AF Hackett, KJ Clark, EA Whitcomb, RF Camp, M Tully, JG TI Amended data on arginine utilization by Spiroplasma species SO INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY LA English DT Article ID CLASS MOLLICUTES; CLASSIFICATION; GROWTH AB Hydrolysis of arginine is a classical diagnostic test for species in the mollicute order Entomoplasmatales. In this paper we report data for arginine utilization by spiroplasmas, as determined by standard methods. In addition, modified methods sere developed for fastidious spiroplasmas, such as strain LD-1(T) (T = type strain), the Colorado potato beetle spiroplasma, Twenty-one spiroplasma strains representing 13 groups or subgroups and eight ungrouped spiroplasmas (seven of which represent putative groups) were studied, The arginine reactions of eight strains were the same as the reactions reported previously, but previously reported positive tests for spiroplasma subgroups I-5 and I-6 (Spiroplasma insolitum) could not be repeated, and the data for the latter taxa are corrected. Although other workers have reported that addition of carbohydrate to media may be necessary for the utilization of arginine, the presence of glucose tended to obscure arginine hydrolysis In our studies. C1 NIAID,MYCOPLASMA SECT,MOL MICROBIOL LAB,FREDERICK CANC RES FACIL,FREDERICK,MD 21701. RP Hackett, KJ (reprint author), USDA ARS,BELTSVILLE AGR RES CTR,INSECT BIOCONTROL LAB,ROOM 214,BLDG 011A,BARC-W,BELTSVILLE,MD 20705, USA. NR 21 TC 8 Z9 8 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0020-7713 J9 INT J SYST BACTERIOL JI Int. J. Syst. Bacteriol. PD OCT PY 1996 VL 46 IS 4 BP 912 EP 915 PG 4 WC Microbiology SC Microbiology GA VL867 UT WOS:A1996VL86700012 ER PT J AU Hackett, KJ Whitcomb, RF French, FE Tully, JG Gasparich, GE Rose, DL Carle, P Bove, JM Henegar, RB Clark, TB Konai, M Clark, EA Williamson, DL AF Hackett, KJ Whitcomb, RF French, FE Tully, JG Gasparich, GE Rose, DL Carle, P Bove, JM Henegar, RB Clark, TB Konai, M Clark, EA Williamson, DL TI Spiroplasma corruscae sp nov, from a firefly beetle (Coleoptera: Lampyridae) and tabanid flies (Diptera: Tabanidae) SO INTERNATIONAL JOURNAL OF SYSTEMATIC BACTERIOLOGY LA English DT Article ID STEROL-REQUIRING MOLLICUTES; FAM-NOV; DEFORMATION; INSECTS; FLOWERS; ECOLOGY AB Spiroplasma strain EC-1(T) (T = type strain), which was isolated from the gut of a lampyrid beetle (Ellychnia corrusca) in Maryland, was serologically distinct from other spiroplasma species and groups. Similar strains were obtained from other E. corrusca specimens, and, later, numerous isolates of similar or partially related strains were obtained from several species of tabanid flies. Cells of strain EC-1(T) were helical, motile filaments that were bound by a single cytoplasmic membrane, and there was no evidence of a cell wall. The cells were filterable through 220-nm-pore-size membrane filters but not through 100-nm-pore-size membrane filters. The organism was absolutely resistant to penicillin (1,000 U/ml) and required sterol for growth. Strain EC-1(T) grew well in M1D and SP-4 liquid media and could be cultivated in the Edward formulation of conventional mycoplasma medium and in 1% serum fraction medium. Optimal growth occurred at 32 degrees C (doubling time, 1.5 h). Strain EC-1(T) multiplied at 10 to 41 degrees C, but not at 5 or 43 degrees C. This organism produced acid from glucose, but did not hydrolyze arginine or utilize urea. The guanine-plus-cytosine content of the DNA was determined to be 26.3 mol% by the melting temperature method and 27.0 mol% by the buoyant density method. As a result of our studies, strain EC-1 (= ATCC 43212) is designated the type strain of a new species, Spiroplasma corruscae. C1 USDA ARS,BELTSVILLE AGR RES CTR,INSECT BIOCONTROL LAB,BELTSVILLE,MD 20705. GEORGIA SO UNIV,DEPT BIOL,STATESBORO,GA 30460. NIAID,MYCOPLASM SECT,MOL MICROBIOL LAB,FREDERICK CANC RES FACIL,FREDERICK,MD 21702. INRA,BIOL CELLULAIRE & MOL LAB,F-33883 VILLENAVE DORNON,FRANCE. SUNY STONY BROOK,DEPT ANAT SCI,STONY BROOK,NY 11794. NR 47 TC 8 Z9 8 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0020-7713 J9 INT J SYST BACTERIOL JI Int. J. Syst. Bacteriol. PD OCT PY 1996 VL 46 IS 4 BP 947 EP 950 PG 4 WC Microbiology SC Microbiology GA VL867 UT WOS:A1996VL86700018 PM 8863421 ER PT J AU Wurtz, RH AF Wurtz, RH TI Vision for the control of movement - The Friedenwald Lecture SO INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE LA English DT Article ID MONKEY SUPERIOR COLLICULUS; SACCADE-RELATED ACTIVITY; PARIETAL ASSOCIATION CORTEX; SUPPLEMENTARY EYE FIELD; ORIENTING GAZE SHIFTS; ALERT RHESUS-MONKEY; HEAD-FREE CAT; TECTORETICULOSPINAL SYSTEM; FUNCTIONAL-PROPERTIES; NEURONAL-ACTIVITY RP Wurtz, RH (reprint author), NEI,SENSORIMOTOR RES LAB,BLDG 49,ROOM 2A50,BETHESDA,MD 20892, USA. NR 57 TC 50 Z9 51 U1 0 U2 0 PU LIPPINCOTT-RAVEN PUBL PI PHILADELPHIA PA 227 EAST WASHINGTON SQ, PHILADELPHIA, PA 19106 SN 0146-0404 J9 INVEST OPHTH VIS SCI JI Invest. Ophthalmol. Vis. Sci. PD OCT PY 1996 VL 37 IS 11 BP 2131 EP 2145 PG 15 WC Ophthalmology SC Ophthalmology GA VK656 UT WOS:A1996VK65600002 ER PT J AU Sartani, G Silver, PB Rizzo, LV Chan, CC Wiggert, B MAstorakos, G Caspi, RR AF Sartani, G Silver, PB Rizzo, LV Chan, CC Wiggert, B MAstorakos, G Caspi, RR TI Anti-tumor necrosis factor alpha therapy suppresses the induction of experimental autoimmune uveoretinitis in mice by inhibiting antigen priming SO INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE LA English DT Article DE antibodies; autoimmune disease; experimental autoimmune uveitis (EAU); mouse; tumor necrosis factor (TNF); uveitis ID MULTIPLE-SCLEROSIS; FACTOR TNF; T-CELLS; LYMPHOTOXIN; CYTOKINES; ADHESION; INVIVO; RAT; ENCEPHALOMYELITIS; PATHOGENESIS AB Purpose. Experimental autoimmune uveoretinitis (EAU) serves as a model for several immune-mediated diseases that affect the eye in humans. Previous studies indicated that tumor necrosis factor alpha (TNF-alpha) has an important proinflammatory role in EAU and possibly in human uveitis. In this study, the authors investigated the effect of anti-TNF-alpha therapy on EAU in mice. Methods. Experimental autoimmune uveoretinitis was induced in B10.A mice by immunization with interphotoreceptor retinoid-binding protein (IRBP). The mice were treated with 100 or 300 mu l rabbit antiserum or polyclonal antibodies to human TNF-alpha. The treatment spanned either the afferent or the efferent stage of EAU (days-1, 1, 3, 5, 7, or days 8, 10, 12, 14, 16, respectively). Control animals were injected with preimmune rabbit serum at the corresponding times or were not treated. Three weeks after immunization, EAU was assessed by clinical evaluation and by histopathology. Immunologic responses were assessed by delayed-type hypersensitivity (DTH), lymphocyte proliferation to IRBP, and relative abundance of IRBP-primed splenocytes. Results. The treatment with rabbit anti-TNF-alpha serum significantly ameliorated disease when given during the afferent stage but had no effect when given during the efferent stage of EAU. The effect on DTH, lymphocyte proliferation, and abundance of antigen-reactive cells roughly paralleled the effect on disease. Conclusions. Neutralization of systemic TNF ameliorates EAU. The effectiveness of afferent treatment in comparison to the treatment during the efferent stage, together with the reduced proliferation and the reduced abundance of IRBP-responsive cells, suggest that interference with afferent-acting processes such as antigen priming is important to achieve protection from EAU by anti-TNF treatment. C1 NEI,IMMUNOL LAB,NIH,BETHESDA,MD 20892. NICHHD,NIH,BETHESDA,MD 20892. RI Rizzo, Luiz Vicente/B-4458-2009 NR 48 TC 71 Z9 73 U1 0 U2 0 PU LIPPINCOTT-RAVEN PUBL PI PHILADELPHIA PA 227 EAST WASHINGTON SQ, PHILADELPHIA, PA 19106 SN 0146-0404 J9 INVEST OPHTH VIS SCI JI Invest. Ophthalmol. Vis. Sci. PD OCT PY 1996 VL 37 IS 11 BP 2211 EP 2218 PG 8 WC Ophthalmology SC Ophthalmology GA VK656 UT WOS:A1996VK65600009 PM 8843907 ER PT J AU Lizak, MJ Mori, K Ceckler, TL Balaban, RS Kador, PF AF Lizak, MJ Mori, K Ceckler, TL Balaban, RS Kador, PF TI Quantitation of galactosemic cataracts in dogs using magnetization transfer contrast-enhanced magnetic resonance imaging SO INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE LA English DT Article DE cataract; dog; lens; magnetic resonance imaging (MRI); magnetization transfer contrast (MTC) ID SUGAR CATARACT; RABBIT LENS; WATER; SPECTROSCOPY; RELAXATION; DIFFUSION; T1 AB Purpose. Magnetic resonance imaging (MRI) is becoming increasingly important for the diagnosis and characterization of ocular pathologies. A drawback to this technique is that image contrast between different regions of tissue can be obscured because of the similarity of their nuclear magnetic resonance relaxation parameters. This problem is addressed by magnetization transfer contrast (MTC) enhancement, a MRI technique that generates high-contrast images based on characteristic tissue differences resulting from the interaction of water and macromolecules. The purpose of this study was to investigate the feasibility-of using MTC-enhanced imaging to monitor quantitatively the lens changes associated with sugar cataract formation in galactose-fed dogs. Methods. Male beagles fed a diet containing 30% galactose were periodically examined by MRI for changes in tissue character. Each examination included a gradient recalled echo image (M(0)), an MTC-enhanced gradient recalled echo image (M(s)), a T-1 image determined from a one-shot T-1 imaging sequence, and a T-1-weighted image taken from the raw T-1 data. Average values were obtained for several regions of interest and tabulated. These were correlated with cataractous stages visually observed by slit lamp biomicroscopy and retroillumination photography. Results. Enhanced image details of the lens and anterior segment that documented osmotic changes from initial cortical vacuole formation to cortical and nuclear changes associated with advanced sugar cataracts were characterized from measurements of parameters obtained from M(0), M(s), T-1-weighted, and T-1 images. Changes in the cross-sectional areas of lenses during sugar cataract formation also were documented. The magnetic resonance images showed visible changes from the onset of cortical vacuole formation. Region of interest (ROI) analysis of the images showed tissue changes occurring throughout the cataract progression. Conclusions. The MTC-enhanced MRI technique is well suited to detecting lens changes associated with cataractogenesis. All but the earliest changes were readily apparent from the images with no further analysis. Graphic ROI analysis was able to detect regional changes associated the cataract progression for all degrees of severity. Furthermore, the images demonstrated changes in size and shape that would not be detectable by visual inspection. C1 NHLBI,CARDIAC ENERGET LAB,NIH,BETHESDA,MD 20892. RP Lizak, MJ (reprint author), NEI,LAB OCULAR THERAPEUT,10 CTR DR,MSC 1850,BETHESDA,MD 20892, USA. RI Balaban, Robert/A-7459-2009 OI Balaban, Robert/0000-0003-4086-0948 NR 17 TC 13 Z9 14 U1 1 U2 1 PU LIPPINCOTT-RAVEN PUBL PI PHILADELPHIA PA 227 EAST WASHINGTON SQ, PHILADELPHIA, PA 19106 SN 0146-0404 J9 INVEST OPHTH VIS SCI JI Invest. Ophthalmol. Vis. Sci. PD OCT PY 1996 VL 37 IS 11 BP 2219 EP 2227 PG 9 WC Ophthalmology SC Ophthalmology GA VK656 UT WOS:A1996VK65600010 PM 8843908 ER PT J AU Miyake, M Hirayama, T Wada, A Nagata, H Makiyama, K Moss, J Kato, I Noda, M AF Miyake, M Hirayama, T Wada, A Nagata, H Makiyama, K Moss, J Kato, I Noda, M TI ADP-ribosyltransferase activity detected in Helicobacter pylori cell extract SO JAPANESE JOURNAL OF MEDICAL SCIENCE & BIOLOGY LA English DT Article C1 NAGASAKI UNIV,INST TROP MED,DEPT BACTERIOL,NAGASAKI 852,JAPAN. NAGASAKI UNIV,DEPT INTERNAL MED 2,NAGASAKI 852,JAPAN. NHLBI,PULM CRIT CARE MED BRANCH,NIH,BETHESDA,MD. RP Miyake, M (reprint author), CHIBA UNIV,SCH MED,DEPT MICROBIOL 2,CHUO KU,CHIBA 260,JAPAN. NR 0 TC 0 Z9 0 U1 0 U2 0 PU NATL INST INFECTIOUS DISEASES PI TOKYO PA C/O JPN J MED SCI BIOL, TOYAMA 1-23-1, SHINJUKU-KU, TOKYO 162, JAPAN SN 0021-5112 J9 JPN J MED SCI BIOL JI Jpn. J. Med. Sci. Biol. PD OCT-DEC PY 1996 VL 49 IS 5-6 BP 253 EP 254 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA WJ360 UT WOS:A1996WJ36000011 ER PT J AU Mishima, K Satoh, K Hishikawa, Y AF Mishima, K Satoh, K Hishikawa, Y TI Effects of melatonin on circadian time keeping system. SO JAPANESE JOURNAL OF NEUROPSYCHOPHARMACOLOGY LA Japanese DT Article ID CORE BODY-TEMPERATURE; BLIND MAN; JET-LAG; SLEEP; HUMANS; LIGHT; SUPPRESSION; ENTRAINMENT; RHYTHMS; RAT C1 AKITA UNIV,SCH MED,DEPT NEUROPSYCHIAT,AKITA 010,JAPAN. NIMH,NCNP,BETHESDA,MD 20892. NR 41 TC 0 Z9 0 U1 0 U2 0 PU SEIWA SHOTEN PUBL LTD PI TOKYO PA 2-5 KAMITAKAIDO 1-CHOME, SUGINAMI-KU, TOKYO 168, JAPAN SN 0388-7588 J9 JPN J NEUROPSYCHOPH JI Jpn. J. Neuropsychopharmacol. PD OCT PY 1996 VL 18 IS 10 BP 711 EP 718 PG 8 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA VW278 UT WOS:A1996VW27800005 ER PT J AU Arnold, LE AF Arnold, LE TI Sex differences in ADHD: Conference summary SO JOURNAL OF ABNORMAL CHILD PSYCHOLOGY LA English DT Article ID ATTENTION-DEFICIT HYPERACTIVITY; TEACHER RATINGS; DISORDER; CHILDREN; GIRLS; CHILDHOOD; BOYS; ADD; GENDER; PEER AB Clinical samples of attention deficit hyperactivity disorder (ADHD) have been dominated by males. Consequently, female manifestations and sex differences have been relatively neglected in the extensive ADHD research. Because ADHD is so common (3% to 5% of school children) and chronic (lifelong in many cases), even a small proportion of females multiplied by such a large base means hundreds of thousands of girls and women with ADHD, a significant public health problem. An NIMH conference concluded that research is needed not only on sex differences related to ADHD, but also on manifestations of ADHD in females as such. Areas of focus should include differences in life course (sex-differential age effects); effects of hormones; effects of ADHD parenting (in utero and postnatal) on the next generation; response to and implications for design of psychosocial treatment; effects of differential comorbidity; normative ''background'' sex differences that influence the manifestation of ADHD; differences in development of verbal fluency and social behavior; possible interactions of sex and ethnicity; a prospective study of both sex offspring of ADHD adults; and such methodological issues as appropriate instruments and diagnostic thresholds, power to prevent false negatives, valid impairment measures, validity and reliability of child self-reports, and more inclusive samples (all three subtypes: inattentive, hyperactive-impulsive, and combined). C1 NIMH,CHILD & ADOLESCENT DISORDERS RES BRANCH,ROCKVILLE,MD 20906. NR 55 TC 151 Z9 154 U1 3 U2 14 PU PLENUM PUBL CORP PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 SN 0091-0627 J9 J ABNORM CHILD PSYCH JI J. Abnorm. Child Psychol. PD OCT PY 1996 VL 24 IS 5 BP 555 EP 569 DI 10.1007/BF01670100 PG 15 WC Psychology, Clinical; Psychology, Developmental SC Psychology GA VW876 UT WOS:A1996VW87600002 PM 8956084 ER PT J AU Dickie, P Mounts, P Purcell, D Miller, G Fredrickson, T Chang, LJ Martin, MA AF Dickie, P Mounts, P Purcell, D Miller, G Fredrickson, T Chang, LJ Martin, MA TI Myopathy and spontaneous Pasteurella pneumotropica-induced abscess formation in an HIV-1 transgenic mouse model SO JOURNAL OF ACQUIRED IMMUNE DEFICIENCY SYNDROMES AND HUMAN RETROVIROLOGY LA English DT Article DE Pasteurella pneumotropica; abscess; HIV-1; mouse model ID HUMAN-IMMUNODEFICIENCY-VIRUS; LONG TERMINAL REPEAT; MURINE IMMUNE-SYSTEM; POLYCLONAL ACTIVATION; NERVOUS-SYSTEM; TAT GENE; MICE; EXPRESSION; CELLS; RETROVIRUS AB In an effort to augment human immunodeficiency virus type 1 (HIV-1) gene expression in transgenic mice, an infectious proviral DNA clone was modified by deleting the two NF kappa B binding sites and some adjacent upstream LTR sequences and replacing them with the core enhancer of Moloney murine leukemia virus (MLV). Two independent lines of MLV/HIV transgenic mice were established that expressed HIV-1-specific RNA in lymphoid tissue, striated skeletal muscle, and the eye lens. Heterozygous animals from each transgenic line spontaneously developed an inflammatory disease of the eye associated with the production of copious amounts of purulent lacrimal secretions beginning at 2 weeks of age. Periorbital abscess formation became grossly apparent by 2 months of age and Pasteurella pneumotropica was cultured from the harderian glands and conjunctival surfaces of many of the MLV/HIV animals but not their nontransgenic, cohabiting littermates. This gram-negative commensal bacterium has been previously associated with a similar disease phenotype in immunocompromised (e.g., nude mice) rodent colonies. MLV/HIV mice developed normally until 15 weeks of age, when weight loss and wasting occurred, culminating in premature death (as earlier as 6 months of age). The cachexia was associated with an initially focal and subsequently progressive myopathy, coinciding with age-related increases of HIV gene expression in muscle. C1 NIAID,MOL MICROBIOL LAB,NIH,BETHESDA,MD 20892. NIH,NATL CTR RES RESOURCES,VET RESOURCES PROGRAM,BETHESDA,MD 20892. NCI,DIV CANC ETIOL,NIH,BETHESDA,MD 20892. MACFARLANE BURNET CTR MED RES,FAIRFIELD,VIC,AUSTRALIA. RI Purcell, Damian/G-5068-2011 NR 41 TC 13 Z9 13 U1 0 U2 2 PU LIPPINCOTT-RAVEN PUBL PI PHILADELPHIA PA 227 EAST WASHINGTON SQ, PHILADELPHIA, PA 19106 SN 1077-9450 J9 J ACQ IMMUN DEF SYND JI J. Acquir. Immune Defic. Syndr. Hum. Retrovirol. PD OCT 1 PY 1996 VL 13 IS 2 BP 101 EP 116 PG 16 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA VL576 UT WOS:A1996VL57600001 PM 8862275 ER PT J AU Flamand, L Zeman, RA Bryant, JL LunardiIskandar, Y Gallo, RC AF Flamand, L Zeman, RA Bryant, JL LunardiIskandar, Y Gallo, RC TI Absence of human herpesvirus 8 DNA sequences in neoplastic Kaposi's sarcoma cell lines SO JOURNAL OF ACQUIRED IMMUNE DEFICIENCY SYNDROMES AND HUMAN RETROVIROLOGY LA English DT Article DE Kaposi's sarcoma; HHV-8; malignancy ID LONG-TERM CULTURE; HOMOSEXUAL MEN; GROWTH-FACTOR; AIDS; INFECTION AB The recent detection of herpesvirus-like DNA sequences in Kaposi's sarcoma (KS) lesions has led to numerous speculations regarding the role of this new agent in KS pathogenesis. However, recent studies indicate a far wider distribution of such viral sequences, shadowing the potential etiologic role of this agent in KS. In this report we show that malignant KS cell lines do not harbor such viral sequences while B cells, CD14(+) and CD34(+) cells do, suggesting that if a KS malignancy originates from infection with HHV-8, the virus can be lost and is not necessary for maintenance of the neoplastic state. Alternatively, HHV-8 may be a ''passenger'' in KS. C1 UNIV MARYLAND,INST BIOTECHNOL,INST HUMAN VIROL,BALTIMORE,MD 21201. NCI,TUMOR CELL BIOL LAB,NIH,BETHESDA,MD 20892. NR 27 TC 42 Z9 43 U1 0 U2 0 PU LIPPINCOTT-RAVEN PUBL PI PHILADELPHIA PA 227 EAST WASHINGTON SQ, PHILADELPHIA, PA 19106 SN 1077-9450 J9 J ACQ IMMUN DEF SYND JI J. Acquir. Immune Defic. Syndr. Hum. Retrovirol. PD OCT 1 PY 1996 VL 13 IS 2 BP 194 EP 197 PG 4 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA VL576 UT WOS:A1996VL57600011 PM 8862285 ER PT J AU Santelli, J Kouzis, A Newcomer, S AF Santelli, J Kouzis, A Newcomer, S TI School-based health centers and adolescent use of primary care and hospital care SO JOURNAL OF ADOLESCENT HEALTH LA English DT Article DE school-based health centers; emergency rooms; hospitalization; primary health care; adolescence ID PROGRAM AB Purpose: Little is known about the impact of school-based primary care on adolescents' use of hospital and emergency room care. Methods: Students (grades 6-12) in nine Baltimore schools with school-based health centers and four schools without health centers were surveyed in May 1991 using an anonymous classroom questionnaire. Self-reported use of primary care services and emergency rooms and hospitalization were examined over the academic year. Logistic regression was used to assess factors influencing use of health care including the presence of a school health center. Results: Students (n = 3,258) in health center schools and comparison schools reported similar rates of chronic health conditions. Students from schools with health centers were more likely to report seeing a social worker or counselor and more likely to report the use of certain health services in the past 4 years. Self-reported emergency room use (38%) and hospitalization (19%) were common. Students in schools with health centers were less likely to report hospitalization (OR = 0.80, 95% CI = 0.66-0.98). Emergency roam use was also lower but only for students attending the school with a health center for more than 1 year (OR = 0.78, 95% CI = 0.62-0.99). Significant predictors of hospital care included reporting one or more chronic health condition, having health insurance, being of African-American race, of older age, and lower grade. Conclusions: Access to school-based, primary health care for adolescents was associated with increased use of primary care, reduced use of emergency rooms, and fewer hospitalizations. These findings have implications for both access to primary care and funding of school-based primary care. C1 JOHNS HOPKINS UNIV,SCH HYG & PUBL HLTH,DEPT MATERNAL & CHILD HLTH,BALTIMORE,MD. JOHNS HOPKINS UNIV,SCH HYG & PUBL HLTH,DEPT MENTAL HYG,BALTIMORE,MD 21218. NICHHD,DEMOG & BEHAV SCI BRANCH,BETHESDA,MD 20892. NR 14 TC 42 Z9 43 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 SN 1054-139X J9 J ADOLESCENT HEALTH JI J. Adolesc. Health PD OCT PY 1996 VL 19 IS 4 BP 267 EP 275 DI 10.1016/S1054-139X(96)00088-2 PG 9 WC Psychology, Developmental; Public, Environmental & Occupational Health; Pediatrics SC Psychology; Public, Environmental & Occupational Health; Pediatrics GA VM699 UT WOS:A1996VM69900005 PM 8897104 ER PT J AU Fishman, ML Rodriguez, L Chau, HK AF Fishman, ML Rodriguez, L Chau, HK TI Molar masses and sizes of starches by high-performance size-exclusion chromatography with on-line multi-angle laser light scattering detection SO JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY LA English DT Article DE starch; molar mass; size; HPSEC; light scattering ID AMYLOSE; AMYLOPECTIN AB Starch was characterized for analysis by high-performance size-exclusion chromatography (HPSEC) with detection by multiple-angle laser light-scattering and refractive index. Corn starches (amylopectin-to-amylose ratios of 1:0, 3:1, 1:1, and 3:7), presolubilized potato starch, and potato starch granules were analyzed. For corn starches, analysis by HPSEC revealed that weight average molar mass (M(w)) and z-average root mean square radius of gyration (R(gz)) decreased with increasing percentage of amylose. For potato starches, granules had a much higher M(w) and R(gz) than presolubilized starch. All chromatograms were polymodal in nature with at least two high M(w) amylopectin components. Except for high amylose corn starch, R(gz) values from this study were comparable with results from an earlier HPSEC/viscometry study. The R(gz) values from microscopic studies were lower than from chromatographic studies. C1 NEI, LAB OCULAR THERAPEUT, BETHESDA, MD 20892 USA. RP Fishman, ML (reprint author), USDA ARS, EASTERN REG RES CTR, 600 E MERMAID LANE, WYNDMOOR, PA 19038 USA. NR 14 TC 55 Z9 57 U1 2 U2 7 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0021-8561 EI 1520-5118 J9 J AGR FOOD CHEM JI J. Agric. Food Chem. PD OCT PY 1996 VL 44 IS 10 BP 3182 EP 3188 DI 10.1021/jf9600162 PG 7 WC Agriculture, Multidisciplinary; Chemistry, Applied; Food Science & Technology SC Agriculture; Chemistry; Food Science & Technology GA VN644 UT WOS:A1996VN64400049 ER PT J AU Joseph, RE Su, TP Cone, EJ AF Joseph, RE Su, TP Cone, EJ TI In vitro binding studies of drugs to hair: Influence of melanin and lipids on cocaine binding to Caucasoid and Africoid hair SO JOURNAL OF ANALYTICAL TOXICOLOGY LA English DT Article ID MASS-SPECTROMETRY; INTEGRAL LIPIDS; IN-VITRO; METABOLITES; BENZOYLECGONINE; IDENTIFICATION; ABUSE C1 NIDA,DIV INTRAMURAL RES,ADDICT RES CTR,BALTIMORE,MD 21224. NR 30 TC 58 Z9 59 U1 0 U2 3 PU PRESTON PUBLICATIONS INC PI NILES PA 7800 MERRIMAC AVE PO BOX 48312, NILES, IL 60648 SN 0146-4760 J9 J ANAL TOXICOL JI J. Anal. Toxicol. PD OCT PY 1996 VL 20 IS 6 BP 338 EP 344 PG 7 WC Chemistry, Analytical; Toxicology SC Chemistry; Toxicology GA VK231 UT WOS:A1996VK23100002 PM 8889667 ER PT J AU Kuhlman, JJ Lalani, S Magluilo, J Levine, B Darwin, WD Johnson, RE Cone, EJ AF Kuhlman, JJ Lalani, S Magluilo, J Levine, B Darwin, WD Johnson, RE Cone, EJ TI Human pharmacokinetics of intravenous, sublingual, and buccal buprenorphine SO JOURNAL OF ANALYTICAL TOXICOLOGY LA English DT Article ID OPIOID DEPENDENCE; HEROIN-ADDICTS; CLINICAL-TRIAL; METHADONE; DETOXIFICATION; PHARMACOLOGY; MAINTENANCE; OPIATE C1 NIDA,DIV INTRAMURAL RES,NIH,BALTIMORE,MD. RP Kuhlman, JJ (reprint author), ARMED FORCES INST PATHOL,DIV FORENS TOXICOL,WASHINGTON,DC 20306, USA. NR 26 TC 137 Z9 139 U1 1 U2 5 PU PRESTON PUBLICATIONS INC PI NILES PA 7800 MERRIMAC AVE PO BOX 48312, NILES, IL 60648 SN 0146-4760 J9 J ANAL TOXICOL JI J. Anal. Toxicol. PD OCT PY 1996 VL 20 IS 6 BP 369 EP 378 PG 10 WC Chemistry, Analytical; Toxicology SC Chemistry; Toxicology GA VK231 UT WOS:A1996VK23100007 PM 8889672 ER PT J AU Cone, EJ Jufer, R Darwin, WD Needleman, SB AF Cone, EJ Jufer, R Darwin, WD Needleman, SB TI Forensic drug testing for opiates .7. Urinary excretion profile of intranasal (snorted) heroin SO JOURNAL OF ANALYTICAL TOXICOLOGY LA English DT Article ID POPPY SEED INGESTION; GAS-CHROMATOGRAPHY; MORPHINE; CODEINE; URINALYSIS; 6-ACETYLMORPHINE; CONSUMPTION C1 USN,DRUG SCREENING LAB,GREAT LAKES,IL 60088. RP Cone, EJ (reprint author), NIDA,ADDICT RES CTR,NIH,POB 5180,BALTIMORE,MD 21224, USA. NR 19 TC 24 Z9 25 U1 0 U2 3 PU PRESTON PUBLICATIONS INC PI NILES PA 7800 MERRIMAC AVE PO BOX 48312, NILES, IL 60648 SN 0146-4760 J9 J ANAL TOXICOL JI J. Anal. Toxicol. PD OCT PY 1996 VL 20 IS 6 BP 379 EP 392 PG 14 WC Chemistry, Analytical; Toxicology SC Chemistry; Toxicology GA VK231 UT WOS:A1996VK23100008 PM 8889673 ER PT J AU Huestis, MA Mitchell, JM Cone, EJ AF Huestis, MA Mitchell, JM Cone, EJ TI Urinary excretion profiles of 11-nor-9-carboxy-Delta(9)-tetrahydrocannabinol in humans after single smoked doses of marijuana SO JOURNAL OF ANALYTICAL TOXICOLOGY LA English DT Article ID GAS-CHROMATOGRAPHY; GC-MS; TETRAHYDROCANNABINOL; METABOLITES; CANNABINOIDS; USERS; DELTA-1-TETRAHYDROCANNABINOL; DISPOSITION C1 USN,AIR STN,NAVY DRUG SCREENING LAB,JACKSONVILLE,FL 32212. RP Huestis, MA (reprint author), NIDA,ADDICT RES CTR,NIH,BALTIMORE,MD 21224, USA. NR 21 TC 69 Z9 75 U1 2 U2 7 PU PRESTON PUBLICATIONS INC PI NILES PA 7800 MERRIMAC AVE PO BOX 48312, NILES, IL 60648 SN 0146-4760 J9 J ANAL TOXICOL JI J. Anal. Toxicol. PD OCT PY 1996 VL 20 IS 6 BP 441 EP 452 PG 12 WC Chemistry, Analytical; Toxicology SC Chemistry; Toxicology GA VK231 UT WOS:A1996VK23100016 PM 8889681 ER PT J AU Oyler, J Darwin, WD Preston, KL Suess, P Cone, EJ AF Oyler, J Darwin, WD Preston, KL Suess, P Cone, EJ TI Cocaine disposition in meconium from newborns of cocaine-abusing mothers and urine of adult drug users SO JOURNAL OF ANALYTICAL TOXICOLOGY LA English DT Article ID CHROMATOGRAPHY MASS-SPECTROMETRY; CLINICAL IMPLICATIONS; COCAETHYLENE; METABOLITES; ETHANOL; INFANTS; PHARMACOKINETICS; BENZOYLECGONINE; IDENTIFICATION; QUANTITATION C1 NIDA,ADDICT RES CTR,NIH,BALTIMORE,MD 21224. RI Preston, Kenzie/J-5830-2013 OI Preston, Kenzie/0000-0003-0603-2479 NR 42 TC 52 Z9 53 U1 0 U2 3 PU PRESTON PUBLICATIONS INC PI NILES PA 7800 MERRIMAC AVE PO BOX 48312, NILES, IL 60648 SN 0146-4760 J9 J ANAL TOXICOL JI J. Anal. Toxicol. PD OCT PY 1996 VL 20 IS 6 BP 453 EP 462 PG 10 WC Chemistry, Analytical; Toxicology SC Chemistry; Toxicology GA VK231 UT WOS:A1996VK23100017 PM 8889682 ER PT J AU Heishman, SJ Singleton, EG Crouch, DJ AF Heishman, SJ Singleton, EG Crouch, DJ TI Laboratory validation study of drug evaluation and classification program: Ethanol, cocaine, and marijuana SO JOURNAL OF ANALYTICAL TOXICOLOGY LA English DT Article ID IONIZATION MASS-SPECTROMETRY; GAS-CHROMATOGRAPHY; MAJOR METABOLITES; PERFORMANCE; ALCOHOL; DRIVERS; HUMANS; ABUSE; URINE C1 JOHNS HOPKINS UNIV,SCH MED,DEPT PSYCHIAT & BEHAV SCI,BALTIMORE,MD 21205. MORGAN STATE UNIV,DEPT PSYCHOL,BALTIMORE,MD 21239. UNIV UTAH,CTR HUMAN TOXICOL,SALT LAKE CITY,UT. RP Heishman, SJ (reprint author), NIDA,ADDICT RES CTR,BIOL DEPENDENCE SECT,POB 5180,BALTIMORE,MD 21221, USA. OI Singleton, Edward G./0000-0003-3442-877X NR 43 TC 19 Z9 19 U1 2 U2 6 PU PRESTON PUBLICATIONS INC PI NILES PA 7800 MERRIMAC AVE PO BOX 48312, NILES, IL 60648 SN 0146-4760 J9 J ANAL TOXICOL JI J. Anal. Toxicol. PD OCT PY 1996 VL 20 IS 6 BP 468 EP 483 PG 16 WC Chemistry, Analytical; Toxicology SC Chemistry; Toxicology GA VK231 UT WOS:A1996VK23100019 PM 8889684 ER PT J AU Nussenblatt, RB Gery, I AF Nussenblatt, RB Gery, I TI Experimental autoimmune uveitis and its relationship to clinical ocular inflammatory disease SO JOURNAL OF AUTOIMMUNITY LA English DT Review ID RETINOID-BINDING-PROTEIN; T-CELL RECEPTOR; PIGMENT EPITHELIAL-CELLS; IMMUNE DEVIATION ACAID; GROWTH-FACTOR-BETA; S-ANTIGEN; ANTERIOR-CHAMBER; LEWIS RATS; AQUEOUS-HUMOR; SYMPATHETIC OPHTHALMIA RP Nussenblatt, RB (reprint author), NEI, IMMUNOL LAB, BLDG 10, ROOM 10 N202, 10 CTR LANE, BETHESDA, MD 20892 USA. NR 131 TC 37 Z9 37 U1 0 U2 0 PU ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0896-8411 J9 J AUTOIMMUN JI J. Autoimmun. PD OCT PY 1996 VL 9 IS 5 BP 575 EP 585 DI 10.1006/jaut.1996.0077 PG 11 WC Immunology SC Immunology GA VQ255 UT WOS:A1996VQ25500001 PM 8933273 ER PT J AU Notkins, AL Lu, J Li, Q VanderVegt, FP Wasserfall, C Maclaren, NK Lan, MS AF Notkins, AL Lu, J Li, Q VanderVegt, FP Wasserfall, C Maclaren, NK Lan, MS TI IA-2 and IA-2 beta are major autoantigens in IDDM and the precursors of the 40 kDa and 37 kDa tryptic fragments SO JOURNAL OF AUTOIMMUNITY LA English DT Article DE autoantigen; islet; IA-2/IA-2 beta; type I diabetes; autoantibodies; 37/40 kDa tryptic fragments ID PROTEIN-TYROSINE-PHOSPHATASE; ISLET-CELL ANTIGEN-512; GLUTAMATE-DECARBOXYLASE; DIABETES-MELLITUS; HUMAN INSULINOMA; IDENTIFICATION; ANTIBODIES; CDNA AB By subtraction strategy and polymerase chain reaction amplification, two novel cDNAs, designated IA-2 and IA-2 beta, were cloned, sequenced and expressed. Both are transmembrane proteins belonging to the protein tyrosine phosphatase family and are expressed in pancreatic islets. Serological studies revealed that a high percentage of patients with IDDM have autoantibodies to IA-2/IA-2 beta and that the presence of these autoantibodies in otherwise normal individuals is highly predictive in identifying those at risk of ultimately developing clinical diabetes. Moreover, many patients who are ICA positive, but who do not have Abs to GAD(65), have Abs to IA-2/IA-2 beta. Enzymatic cleavage of IA-2/IA-2 beta and serological analysis showed that IA-2 is the precursor of the 40 kDa tryptic fragment and IA-2 beta is the precursor of the 37 kDa tryptic fragment, both previously shown to be autoantigens. It is concluded that IA-2/IA-2 beta are major autoantigens in IDDM and together with GAD(65) are responsible for much of the reactivity of ICA with pancreatic islets. Tests for the detection of autoantibodies to recombinant IA-2/IA-2 beta and recombinant GAD(65) are likely to replace the ICA immunofluorescence test for population screening. (C) 1996 Academic Press Limited C1 UNIV FLORIDA,COLL MED,DEPT PATHOL & LAB MED,GAINESVILLE,FL 32610. RP Notkins, AL (reprint author), NIDR,ORAL MED LAB,NIH,30 CONVENT DR,MSC 4322,BLDG 30,ROOM 721,BETHESDA,MD 20892, USA. NR 18 TC 25 Z9 25 U1 0 U2 2 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON, ENGLAND NW1 7DX SN 0896-8411 J9 J AUTOIMMUN JI J. Autoimmun. PD OCT PY 1996 VL 9 IS 5 BP 677 EP 682 DI 10.1006/jaut.1996.0088 PG 6 WC Immunology SC Immunology GA VQ255 UT WOS:A1996VQ25500012 PM 8933284 ER PT J AU Lunsford, RD London, J AF Lunsford, RD London, J TI Natural genetic transformation in Streptococcus gordonii: comX imparts spontaneous competence on strain wicky SO JOURNAL OF BACTERIOLOGY LA English DT Article ID MOLECULAR-CLONING; CONTROL LOCUS; PNEUMONIAE; SYSTEM; SEQUENCES; PROTEINS; FAMILY; MUTANS AB Streptococcus gordonii Wicky becomes competent only after stimulation with conditioned medium from strain Challis as a source of competence factor (CF). A 3.2-kbp genomic fragment from Challis was found to impart spontaneous competence on Wicky by a complementation assay. Wicky clones containing the fragment secreted a heat-sensitive activity that induced competence in Wicky and in a comA insertion mutant of Challis. Activity was localized to a putative open reading frame, comX, with the potential to encode a 52-amino-acid peptide. comX had no similarity to known sequences, and a comX::ermAM insertion mutant of Challis transformed normally and secreted CF. These data suggest that a CF-independent pathway for competence induction exists in S. gordonii. RP Lunsford, RD (reprint author), NIDR,NIH,MICROBIAL ECOL LAB,30 CONVENT DR MSC 4350,BETHESDA,MD 20892, USA. NR 24 TC 38 Z9 41 U1 1 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0021-9193 J9 J BACTERIOL JI J. Bacteriol. PD OCT PY 1996 VL 178 IS 19 BP 5831 EP 5835 PG 5 WC Microbiology SC Microbiology GA VK789 UT WOS:A1996VK78900043 PM 8824638 ER PT J AU Rosa, P Samuels, DS Hogan, D Stevenson, B Casjens, S Tilly, K AF Rosa, P Samuels, DS Hogan, D Stevenson, B Casjens, S Tilly, K TI Directed insertion of a selectable marker into a circular plasmid of Borrelia burgdorferi SO JOURNAL OF BACTERIOLOGY LA English DT Article ID LYME-DISEASE AGENT; OUTER SURFACE PROTEIN; POLYMERASE CHAIN-REACTION; BACILLUS-SUBTILIS; LINEAR PLASMIDS; ENZYMATIC AMPLIFICATION; ANTIGENIC VARIATION; TRANSPORT-SYSTEM; CAUSATIVE AGENT; ETIOLOGIC AGENT AB Studies of the biology of Borrelia burgdorferi and the pathogenesis of Lyme disease are severely limited by the current lack of genetic tools. As an initial step toward facile genetic manipulation of this pathogenic spirochete, we have investigated gene inactivation by allelic exchange using a mutated borrelial gyrB gene that confers resistance to the antibiotic coumermycin A(1) as a selectable marker. We have transformed B. burgdorferi by electroporation with a linear fragment of DNA in which this selectable marker was flanked by sequences from a native borrelial 26-kb circular plasmid. We have identified coumermycin A(1)-resistant transformants in which gyrB had interrupted the targeted site on the 26-kb plasmid via homologous recombination with the flanking sequences. Antibiotic resistance conferred by the mutated gyrB gene on the plasmid is dominant, and transformed spirochetes carrying this plasmid do not contain any unaltered copies of the plasmid. Coumermycin A(1) resistance can be transferred to naive B. burgdorferi by transformation with borrelial plasmid DNA from the initial transformants. This work represents the first example of a directed mutation in B. burgdorferi whereby a large segment of heterologous DNA (gyrB) has been inserted via homologous recombination with flanking sequences, thus demonstrating the feasibility of specific gene inactivation by allelic exchange. C1 UNIV MONTANA,DIV BIOL SCI,MISSOULA,MT 59812. UNIV UTAH,MED CTR,DEPT ONCOL SCI,DIV MOL BIOL & GENET,SALT LAKE CITY,UT 84132. RP Rosa, P (reprint author), NIAID,ROCKY MT LABS,MICROBIAL STRUCT & FUNCT LAB,903 S 4TH ST,HAMILTON,MT 59840, USA. RI Samuels, D Scott/B-7549-2012 OI Samuels, D Scott/0000-0001-8352-7593 NR 61 TC 63 Z9 64 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0021-9193 J9 J BACTERIOL JI J. Bacteriol. PD OCT PY 1996 VL 178 IS 20 BP 5946 EP 5953 PG 8 WC Microbiology SC Microbiology GA VL472 UT WOS:A1996VL47200014 PM 8830691 ER PT J AU Tycko, R AF Tycko, R TI Prospects for resonance assignments in multidimensional solid-state NMR spectra of uniformly labeled proteins SO JOURNAL OF BIOMOLECULAR NMR LA English DT Article DE solid-state NMR; magic angle spinning; chemical shift correlation; resonance assignment ID NUCLEAR-MAGNETIC-RESONANCE; ECHO DOUBLE-RESONANCE; ANGLE-SPINNING NMR; DIPOLAR CORRELATION NMR; ROTATING SOLIDS; CHEMICAL-SHIFTS; CORRELATION SPECTROSCOPY; AB-INITIO; ORGANIC-SOLIDS; QUANTUM-WELLS AB The feasibility of assigning the backbone N-15 and C-13 NMR chemical shifts in multidimensional magic angle spinning NMR spectra of uniformly isotopically labeled proteins and peptides in unoriented solid samples is assessed by means of numerical simulations. The goal of these simulations is to examine how the upper limit on the size of a peptide for which unique assignments can be made depends on the spectral resolution, i.e., the NMR line widths. Sets of simulated three-dimensional chemical shift correlation spectra for artificial peptides of varying length are constructed from published liquid-state NMR chemical shift data for ubiquitin, a well-characterized soluble protein. Resonance assignments consistent with these spectra to within the assumed spectral resolution are found by a numerical search algorithm. The dependence of the number of consistent assignments on the assumed spectral resolution and on the length of the peptide is reported. If only three-dimensional chemical shift correlation data for backbone N-15 and C-13 nuclei are used, and no residue-specific chemical shift information, information from amino acid side-chain signals, and proton chemical shift information are available, a spectral resolution of 1 ppm or less is generally required for a unique assignment of backbone chemical shifts for a peptide of 30 amino acid residues. RP Tycko, R (reprint author), NIDDKD,CHEM PHYS LAB,NIH,5 CTR DR,BETHESDA,MD 20892, USA. NR 49 TC 46 Z9 46 U1 1 U2 6 PU ESCOM SCI PUBL BV PI LEIDEN PA PO BOX 214, 2300 AE LEIDEN, NETHERLANDS SN 0925-2738 J9 J BIOMOL NMR JI J. Biomol. NMR PD OCT PY 1996 VL 8 IS 3 BP 239 EP 251 DI 10.1007/BF00410323 PG 13 WC Biochemistry & Molecular Biology; Spectroscopy SC Biochemistry & Molecular Biology; Spectroscopy GA VW192 UT WOS:A1996VW19200002 PM 8953215 ER PT J AU Tjandra, N Wingfield, P Stahl, S Bax, A AF Tjandra, N Wingfield, P Stahl, S Bax, A TI Anisotropic rotational diffusion of perdeuterated HIV protease from N-15 NMR relaxation measurements at two magnetic SO JOURNAL OF BIOMOLECULAR NMR LA English DT Article DE N-15 NMR; rotational diffusion; chemical shift anisotropy; dynamics; magnetic field dependence; deuteration ID MODEL-FREE APPROACH; BACKBONE DYNAMICS; STAPHYLOCOCCAL NUCLEASE; RESONANCE RELAXATION; CROSS-CORRELATION; SPECTROSCOPY; MACROMOLECULES; PROTEINS; DIPOLAR; RANGE AB N-15 NMR relaxation times in perdeuterated HIV-1 protease, complexed with the sub-nanomolar inhibitor DMP323, have been measured at 600 and 360 MHz H-1 frequency. The relative magnitudes of the principal components of the inertia tensor, calculated from the X-ray coordinates of the protein-drug complex, are 1.0:0.85:0.44. The relation between the T-1/T-2 ratios observed for the individual backbone amides and their N-H orientation within the 3D structure of the protease dimer yields a rotational diffusion tensor oriented nearly collinear to the inertia tenser. The relative magnitudes of its principal components (1.00:1.11:1.42) are also in good agreement with hydrodynamic modeling results. The orientation and magnitude of the diffusion tensors derived from relaxation data obtained at 360 and 600 MHz are nearly identical. The anisotropic nature of the rotational diffusion has little influence on the order parameters derived from the N-15 T-1 and T-2 relaxation times; however, if anisotropy is ignored, this can result in erroneous identification of either exchange broadening or internal motions on a nanosecond time scale. The average ratio of the T-1 values measured at 360 and 600 MHz is 0.50+/-0.015, which is slightly larger than the value of 0.466 expected for an isotropic rigid rotor with tau(c)=10.7 ns. The average ratio of the T-2 values measured at 360 and 600 MHz is 1.14+/-0.04, which is also slightly larger than the expected ratio of 1.11, This magnetic field dependence of the T-1 and T-2 relaxation times suggests that the spectral density contribution from fast internal motions is not negligible, and that the chemical shift anisotropy of peptide backbone amides, on average, is larger than the 160 ppm value commonly used in N-15 relaxation studies of proteins. C1 NIAMSD,PROT EXPRESS LAB,NIH,BETHESDA,MD 20892. RP Tjandra, N (reprint author), NIDDKD,CHEM PHYS LAB,NIH,BLDG 2,BETHESDA,MD 20892, USA. NR 39 TC 201 Z9 201 U1 0 U2 6 PU ESCOM SCI PUBL BV PI LEIDEN PA PO BOX 214, 2300 AE LEIDEN, NETHERLANDS SN 0925-2738 J9 J BIOMOL NMR JI J. Biomol. NMR PD OCT PY 1996 VL 8 IS 3 BP 273 EP 284 DI 10.1007/BF00410326 PG 12 WC Biochemistry & Molecular Biology; Spectroscopy SC Biochemistry & Molecular Biology; Spectroscopy GA VW192 UT WOS:A1996VW19200005 PM 8953218 ER PT J AU Blumenthal, R Sarkar, DP Durell, S Howard, DE Morris, SJ AF Blumenthal, R Sarkar, DP Durell, S Howard, DE Morris, SJ TI Dilation of the influenza hemagglutinin fusion pore revealed by the kinetics of individual cell-cell fusion events SO JOURNAL OF CELL BIOLOGY LA English DT Article ID VIRAL ENVELOPE PROTEIN; INDUCED MEMBRANE-FUSION; RED BLOOD-CELLS; NEUROTRANSMITTER RELEASE; SURFACE-DENSITY; INITIAL-STAGES; MAST-CELLS; VIRUS; FLUORESCENCE; EXOCYTOSIS AB We have monitored kinetics of fusion between cell pairs consisting of a single influenza hemagglutinin (HA)-expressing cell and a single erythrocyte (RBC) that had been labeled with both a fluorescent lipid (DiI) in the membrane and a fluorescent solute (calcein) in the aqueous space. Initial fusion pore opening between the RBC and HA-expressing cell produced a change in RBC membrane potential (Delta psi) that was monitored by a decrease in DiI fluorescence. This event was followed by two distinct stages of fusion pore dilation: the flux of fluorescent lipid (phi(L)) and the flux of a large aqueous fluorescent dye (phi(S)). We have analyzed the kinetics of events that occur as a result of transitions between a fusion pore (FP) and a solute permissive fusion pore (FPS). Our data are consistent with a fusion pore comprising six HA trimers. C1 NCI,MOL STRUCT SECT,DIV BASIC SCI,NIH,BETHESDA,MD 20892. UNIV DELHI,DEPT BIOCHEM,NEW DELHI,INDIA. UNIV MISSOURI,SCH BIOL SCI,DIV MOL BIOL & BIOCHEM,KANSAS CITY,MO 64110. RP Blumenthal, R (reprint author), NCI,SECT MEMBRANE STRUCT & FUNCT,DIV BASIC SCI,NIH,10 CTR DR,MSC 1350,BLDG 10 ROOM 4A01,BETHESDA,MD 20892, USA. NR 49 TC 186 Z9 188 U1 2 U2 6 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 222 E 70TH STREET, NEW YORK, NY 10021 SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD OCT PY 1996 VL 135 IS 1 BP 63 EP 71 DI 10.1083/jcb.135.1.63 PG 9 WC Cell Biology SC Cell Biology GA VM028 UT WOS:A1996VM02800007 PM 8858163 ER PT J AU Guinet, F Dvorak, JA Fujioka, H Keister, DB Muratova, O Kaslow, DC Aikawa, M Vaidya, AB Wellems, TE AF Guinet, F Dvorak, JA Fujioka, H Keister, DB Muratova, O Kaslow, DC Aikawa, M Vaidya, AB Wellems, TE TI A developmental defect in Plasmodium falciparum male gametogenesis SO JOURNAL OF CELL BIOLOGY LA English DT Article ID TRANSMISSION-BLOCKING IMMUNITY; ALPHA-TUBULIN-II; MALARIA PARASITES; GENES; GAMETOCYTES; EXPRESSION; RESISTANCE; INVITRO; PROTEIN; GAMETOCYTOGENESIS AB Asexually replicating populations of Plasmodium parasites, including those from cloned lines, generate both male and female gametes to complete the malaria life cycle through the mosquito. The generation of these sexual forms begins with the induction of gametocytes from haploid asexual stage parasites in the blood of the vertebrate host. The molecular processes that govern the differentiation and development of the sexual forms are largely unknown. Here we describe a defect that affects the development of competent male gametocytes from a mutant clone of P. falciparum (Dd2). Comparison of the Dd2 clone to the predecessor clone from which it was derived (W2'82) shows that the defect is a mutation that arose during the long-term cultivation of asexual stages in vitro. Light and electron microscopic images, and indirect immunofluorescence assays with male-specific anti-alpha-tubulin II antibodies, indicate a global disruption of male development at the gametocyte level, with at least a 70-90% reduction in the proportion of mature male gametocytes by the Dd2 clone relative to W2'82. A high prevalence of abnormal gametocyte forms, frequently containing multiple and unusually large vacuoles, is associated with the defect. The reduced production of mature male gametocytes may reflect a problem in processes that commit a gametocyte to male development or a progressive attrition of viable male gametocytes during maturation. The defect is genetically linked to an almost complete absence of male gamete production and of infectivity to mosquitoes. This is the first sex-specific developmental mutation identified and characterized in Plasmodium. C1 NIAID,PARASIT DIS LAB,NIH,BETHESDA,MD 20892. CASE WESTERN RESERVE UNIV,INST PATHOL,CLEVELAND,OH 44106. MED COLL PENN & HAHNEMANN UNIV,PHILADELPHIA,PA 19102. TOKAI UNIV,SCH MED,RES INST MED SCI,ISEHARA,KANAGAWA 25911,JAPAN. FU NIAID NIH HHS [AI-35827, R01 AI028398] NR 45 TC 37 Z9 38 U1 0 U2 3 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 222 E 70TH STREET, NEW YORK, NY 10021 SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD OCT PY 1996 VL 135 IS 1 BP 269 EP 278 DI 10.1083/jcb.135.1.269 PG 10 WC Cell Biology SC Cell Biology GA VM028 UT WOS:A1996VM02800023 PM 8858179 ER PT J AU Marks, MS Woodruff, L Ohno, H Bonifacino, JS AF Marks, MS Woodruff, L Ohno, H Bonifacino, JS TI Protein targeting by tyrosine- and di-leucine-based signals: Evidence for distinct saturable components SO JOURNAL OF CELL BIOLOGY LA English DT Article ID EPIDERMAL GROWTH-FACTOR; TRANS-GOLGI NETWORK; LOW-DENSITY-LIPOPROTEIN; LYSOSOMAL MEMBRANE GLYCOPROTEIN; MANNOSE 6-PHOSPHATE RECEPTOR; INVARIANT CHAIN COMPLEXES; CELL ANTIGEN RECEPTOR; COATED PIT ADAPTINS; CLASS-II MOLECULES; CYTOPLASMIC DOMAIN AB Targeting of transmembrane proteins to lysosomes, endosomal compartments, or the trans-Golgi network is largely dependent upon cytoplasmically exposed sorting signals. Among the most widely used signals are those that conform to the tyrosine-based motif, YXXempty set (where Y is tyrosine, X is any amino acid, and empty set is an amino acid with a bulky hydrophobic group), and to the di-leucine (or LL) motif. Signals conforming to both motifs have been implicated in protein localization to similar post-Golgi compartments. We have exploited the saturability of sorting to ask whether different YXXempty set or LL signals use shared components of the targeting machinery. Chimeric proteins containing various cytoplasmic domains indoor targeting signals were overexpressed in HeLa cells by transient transfection. Endogenous transferrin receptor and lysosomal proteins accumulated at the cell surface upon overexpression of chimeric proteins containing functional YXXempty set targeting signals, regardless of the compartmental destination imparted by the signal. Furthermore, overexpression of these chimeric proteins compromised YXXempty set-mediated endocytosis and lysosomal delivery. These activities were ablated by mutating the signals or by appending sequences that conformed to the YXXempty set motif but lacked targeting activity. Interestingly, overexpression of chimeric proteins containing cytoplasmic LL signals failed to induce surface displacement of endogenous YXXempty set-containing proteins, but did displace other proteins containing LL motifs. Our data demonstrate that: (a) Protein targeting and internalization mediated by either YXXempty set or LL motifs are saturable processes; (b) common saturable components are used in YXXempty set-mediated protein internalization and targeting to different post-Golgi compartments; and (c) YXXempty set-and LL-mediated targeting mechanisms use distinct saturable components. C1 NICHHD,CELL BIOL & METAB BRANCH,NIH,BETHESDA,MD 20892. UNIV PENN,SCH MED,DEPT PATHOL & LAB MED,PHILADELPHIA,PA 19104. RI Ohno, Hiroshi/L-7899-2014; OI Ohno, Hiroshi/0000-0001-8776-9661; Marks, Michael/0000-0001-7435-7262; Bonifacino, Juan S./0000-0002-5673-6370 NR 80 TC 251 Z9 256 U1 0 U2 4 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 222 E 70TH STREET, NEW YORK, NY 10021 SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD OCT PY 1996 VL 135 IS 2 BP 341 EP 354 DI 10.1083/jcb.135.2.341 PG 14 WC Cell Biology SC Cell Biology GA VP229 UT WOS:A1996VP22900005 PM 8896593 ER PT J AU Muresan, V Godek, CP Reese, TS Schnapp, BJ AF Muresan, V Godek, CP Reese, TS Schnapp, BJ TI Plus-end motors override minus-end motors during transport of squid axon vesicles on microtubules SO JOURNAL OF CELL BIOLOGY LA English DT Article ID MEMBRANOUS ORGANELLES INVIVO; SINGLE KINESIN MOLECULES; CYTOPLASMIC DYNEIN; HEAVY-CHAIN; GIANT-AXON; SYNAPTIC VESICLES; ANTEROGRADE MOTOR; MONOMERIC MOTOR; SOLUBLE FACTORS; PROTEIN AB Plus- and minus-end vesicle populations from squid axoplasm were isolated from each other by selective extraction of the minus-end vesicle motor followed by 5'-adenylyl imidodiphosphate (AMP-PNP)induced microtubule affinity purification of the plus-end vesicles. In the presence of cytosol containing both plus- and minus-end motors, the isolated populations moved strictly in opposite directions along microtubules in vitro. Remarkably, when treated with trypsin before incubation with cytosol, purified plus-end vesicles moved exclusively to microtubule minus ends instead of moving in the normal plus-end direction. This reversal in the direction of movement of trypsinized plus-end vesicles, in light of further observation that cytosol promotes primarily minus-end movement of liposomes, suggests that the machinery for cytoplasmic dynein-driven, minus-end vesicle movement can establish a functional interaction with the lipid bilayers of both vesicle populations. The additional finding that kinesin overrides cytoplasmic dynein when both are bound to bead surfaces indicates that the direction of vesicle movement could be regulated simply by the presence or absence of a tightly bound, plus-end kinesin motor; being processive and tightly bound, the kinesin motor would override the activity of cytoplasmic dynein because the latter is weakly bound to vesicles and less processive. In support of this model, it was found that (a) only plus-end vesicles copurified with tightly bound kinesin motors; and (b) both plus- and minus-end vesicles bound cytoplasmic dynein from cytosol. C1 HARVARD UNIV,SCH MED,DEPT CELL BIOL,BOSTON,MA 02115. NINCDS,NEUROBIOL LAB,NIH,BETHESDA,MD 20892. FU NINDS NIH HHS [NS-26846] NR 73 TC 59 Z9 59 U1 1 U2 1 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 222 E 70TH STREET, NEW YORK, NY 10021 SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD OCT PY 1996 VL 135 IS 2 BP 383 EP 397 DI 10.1083/jcb.135.2.383 PG 15 WC Cell Biology SC Cell Biology GA VP229 UT WOS:A1996VP22900008 PM 8896596 ER PT J AU Schnaper, HW Kopp, JB Poncelet, AC Hubchak, SC StetlerStevenson, WG Klotman, PE Kleinman, HK AF Schnaper, HW Kopp, JB Poncelet, AC Hubchak, SC StetlerStevenson, WG Klotman, PE Kleinman, HK TI Increased expression of extracellular matrix proteins and decreased expression of matrix proteases after serial passage of glomerular mesangial cells SO JOURNAL OF CELL SCIENCE LA English DT Article DE glomerulosclerosis; mesangial cell; collagen; extracellular matrix; extracellular matrix protease ID PLASMINOGEN-ACTIVATOR INHIBITOR; IV COLLAGENASE; TISSUE INHIBITOR; CDNA CLONING; GLOMERULOSCLEROSIS; LAMININ; RAT; METALLOPROTEINASES; FIBROBLASTS; SCLEROSIS AB The cellular events causing pathological extracellular matrix (ECM) accumulation in vivo are not well understood, Prolonged serial passage of several cell types in culture leads to increased production of extracellular matrix (ECM) proteins, but the mechanism for these putative fibrotic changes is not known. Here, human fetal glomerular mesangial cells were subjected to serial passage (P) in culture and the expression of ECM proteins, proteases and protease inhibitors was comprehensively evaluated, From P11 through P14, a series of phenotypic changes occurred. Steady-state expression of mRNA for alpha 1 chains of type III and type IV (but not type I) collagen, and for laminin beta 1 and gamma 1, increased 2- to 8-fold, while expression of mRNA for interstitial collagenase (MMP-1) and gelatinase A (MMP-2) virtually ceased, Expression of tissue-type plasminogen activator (tPA) mRNA also decreased markedly, Expression of mRNA for the tissue inhibitor of metalloproteinases (TIMP)-1, and of the smaller of two mRNA species for the PA inhibitor PAI-1, ceased by P14, There was a switch in expression of the two species of TIMP-2 mRNA: whereas the ratio of signal intensity comparing the 3.5 kb mRNA species to the 1.0 lib species was 5:1 up to P11, it was reversed (1:5) at P14 and later. Serial passage also led to changes in protein expression, with increased type IV collagen and laminin, but decreased interstitial collagenase and gelatinase A. The cells showed a progressive increase in staining for type IV collagen. These findings define the appearance of a matrix-accumulating phenotype in later-passage mesangial cells, Matrix expansion in vivo has been associated with increased transforming growth factor (TGF)-beta synthesis; the cells were found to show at least 5-fold increased expression of TGF-beta 1 mRNA from P8 to P16. However, treatment of P9 or P10 cells with graded doses of TGF-beta 1 increased expression of both collagen IV and gelatinase A mRNA and did not alter the ratio of signal intensity for TIMP-2 mRNA species. Thus, assumption of a matrix-accumulating phenotype by these cultured fetal glomerular mesangial cells is not accelerated by exogenous TGF-beta, These data describe an in vitro model of mesangial cell matrix turnover in which matrix accumulation could result from a concerted increase in ECM synthesis and decrease in ECM degradation. C1 NIDR,DEV BIOL LAB,BETHESDA,MD 20892. NIDDK,METAB DIS BRANCH,BETHESDA,MD. NCI,PATHOL LAB,BETHESDA,MD 20892. MT SINAI HOSP,DEPT MED,NEW YORK,NY 10029. RP Schnaper, HW (reprint author), NORTHWESTERN UNIV,SCH MED,DEPT PEDIAT,303 E CHICAGO AVE,CHICAGO,IL 60611, USA. RI Stetler-Stevenson, William/H-6956-2012; OI Stetler-Stevenson, William/0000-0002-5500-5808; Kopp, Jeffrey/0000-0001-9052-186X FU NIDDK NIH HHS [R01 DK049362, R01-DK49362] NR 46 TC 50 Z9 53 U1 0 U2 0 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE, CAMBS, ENGLAND CB4 4DL SN 0021-9533 J9 J CELL SCI JI J. Cell Sci. PD OCT PY 1996 VL 109 BP 2521 EP 2528 PN 10 PG 8 WC Cell Biology SC Cell Biology GA VQ251 UT WOS:A1996VQ25100011 PM 8923213 ER PT J AU Pompetti, F Rizzo, P Simon, RM Freidlin, B Mew, DJ Pass, HI Picci, P Levine, AS Carbone, M AF Pompetti, F Rizzo, P Simon, RM Freidlin, B Mew, DJ Pass, HI Picci, P Levine, AS Carbone, M TI Oncogene alterations in primary, recurrent, and metastatic human bone tumors SO JOURNAL OF CELLULAR BIOCHEMISTRY LA English DT Article DE osteosarcoma; chondrosarcoma; GCT; oncogene alterations ID SOFT-TISSUE SARCOMAS; T-DELETION MUTANTS; P53 GENE; RETINOBLASTOMA GENE; OSTEO-SARCOMA; BREAST-CANCER; EXPRESSION; AMPLIFICATION; MUTATION; OSTEOSARCOMAS AB We investigated the structure and the expression of various oncogenes in three of the most common human bone tumors-osteosarcoma (36 samples from 34 patients), giant cell tumor (10 patients), and chondrosarcoma (18 patients)-in an attempt to identify the genetic alterations associated with these malignancies. Alterations of RE and p53 were detected only in osteosarcomas. Alterations of c-myc, N-myc, and c-fos were detected in osteosarcomas and giant cell tumors. Ras alterations (H-ras, Ki-ras, N-ras) were rare. Chondrosarcomas did not contain any detectable genetic alterations. Our results suggest that alterations of c-myc, N-myc, and c-fos oncogenes occur in osteosarcomas, in addition to those previously described for the tumor suppressor genes RE and p53. Moreover, statistical analyses indicate that c-fos alterations occur more frequently in osteosarcoma patients with recurrent or metastatic disease. (C) 1996 Wiley-Liss, Inc. C1 UNIV CHICAGO,DEPT PATHOL,CHICAGO,IL 60637. NICHHD,SECT DNA REPLICAT REPAIR & MUTAGENESIS,BETHESDA,MD 20892. NCI,BIOMETR RES BRANCH,DIV CANC TREATMENT,BETHESDA,MD 20892. EMMES CORP,BETHESDA,MD 20892. NCI,THORAC ONCOL SECT,BETHESDA,MD 20892. IST ORTOPEDICI RIZZOLI,LAB ONCOL,I-40121 BOLOGNA,ITALY. RI Picci, Piero/J-5979-2016 OI Picci, Piero/0000-0002-8519-4101 NR 39 TC 84 Z9 89 U1 0 U2 2 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC 605 THIRD AVE, NEW YORK, NY 10158-0012 SN 0730-2312 J9 J CELL BIOCHEM JI J. Cell. Biochem. PD OCT PY 1996 VL 63 IS 1 BP 37 EP 50 PG 14 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA VK547 UT WOS:A1996VK54700003 PM 8891902 ER PT J AU Borre, A Cultraro, CM Segal, S AF Borre, A Cultraro, CM Segal, S TI C-Myc inactivation by mutant Max alters growth and morphology of NCI-H-630 colon cancer cells SO JOURNAL OF CELLULAR PHYSIOLOGY LA English DT Article ID MURINE ERYTHROLEUKEMIA-CELLS; DNA-BINDING; TRANSCRIPTIONAL ACTIVATION; ERYTHROID DIFFERENTIATION; EXTRACELLULAR-MATRIX; CYCLE PROGRESSION; EXPRESSION; PROTEIN; P21; INHIBITION AB The myc gene family has been implicated in multiple cell processes including proliferation, differentiation, tumorigenesis, and apoptosis. For its cellular growth promoting function, Myc must heterodimerize with Max. To study the effect of Myc inactivation on the growth and differentiation properties of epithelial tumor cells, we transfected the H-630 human colon cancer cell line with bm-max; a mutant Max protein in which DNA-binding activity has been abolished. Cells expressing high levels of bm-Max grow poorly, and the morphology of both colonies and single cells is altered. Moreover, increased bm-Max Expression results in a prolonged G(0)/G(1) phase accompanied by induced expression of p21 (WAF1/CIP1), elevated levels of alkaline phosphatase (ALP) activity, and accumulation of large far granuli within the cells. These distinctive cell characteristics are associated with differentiation processes in numerous malignant cell lines. The results of this study support a model in which sequestering of endogenous Myc and Max proteins into ''basic mutant'' dimers lacking DNA-binding activity is sufficient both to inhibit proliferation and to induce changes in cell behavior consistent with differentiation. (C) 1996 Wiley-Liss, Inc.** C1 BETHESDA NAVAL HOSP,NCI,NAVY MED ONCOL BRANCH,BETHESDA,MD 20889. NIH,BETHESDA,MD 20889. UNIFORMED SERV UNIV HLTH SCI,BETHESDA,MD 20889. NR 67 TC 7 Z9 7 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC 605 THIRD AVE, NEW YORK, NY 10158-0012 SN 0021-9541 J9 J CELL PHYSIOL JI J. Cell. Physiol. PD OCT PY 1996 VL 169 IS 1 BP 200 EP 208 PG 9 WC Cell Biology; Physiology SC Cell Biology; Physiology GA VJ730 UT WOS:A1996VJ73000020 PM 8841436 ER PT J AU Laue, L Merke, DP Jones, JV Barnes, KM Hill, S Cutler, GB AF Laue, L Merke, DP Jones, JV Barnes, KM Hill, S Cutler, GB TI A preliminary study of flutamide, testolactone, and reduced hydrocortisone dose in the treatment of congenital adrenal hyperplasia SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID 21-HYDROXYLASE DEFICIENCY; RADIOIMMUNOASSAY; ADOLESCENCE; ESTROGEN; MUTATION; CORTISOL AB Treatment outcome in congenital adrenal hyperplasia is often suboptimal due to hyperandrogenism, treatment-induced hypercortisolism, or both. As a new approach, we hypothesized that the effects of androgen could be blocked by an antiandrogen (flutamide) and an inhibitor of androgen to estrogen conversion (testolactone), thus allowing the hydrocortisone dose to be reduced. We conducted a short term pilot study in 12 children with congenital adrenal hyperplasia in a randomized cross-over open design to determine whether flutamide, testolactone, reduced hydrocortisone dose, and fludrocortisone are more effective than hydrocortisone and fludrcortisone treatment in normalizing linear growth, weight gain, and bone maturation. Each regimen was administered for 6 months, with a 3-month washout period, consisting of hydrocortisone and fludrocortisone treatment, between regimens. Compared to hydrocortisone and fludrocortisone treatment, the regimen of flutamide, testolactone, reduced hydrocortisone dose (from 12.9 to 7.9 mg/m(2) . day), and fludrocortisone produced an increase in plasma 17-hydroxyprogesterone levels (P < 0.05) and a decline in urinary cortisol (P < 0.01), Linear growth rate (-0.9 +/- 0.5 vs. 1.4 +/- 0.6 SD U; P = 0.003), weight velocity (-0.80 +/- 0.4 vs. 0.6 +/- 0.4 SD U; P = 0.01), and bone maturation (0.6 +/- 0.6 vs. 1.4 +/- 0.9 yr bone age/yr chronological age; P = 0.02). Although no important adverse effects were observed, the known potential for flutamide-induced hepatotoxicity made frequent monitoring essential. We conclude that the regimen of flutamide, testolactone, reduced hydrocortisone dose, and fludrocortisone improves the short term control of growth and bone maturation in children with congenital adrenal hyperplasia. Long term studies are required to determine whether this approach can improve these children's growth and development. C1 NICHHD, DEV ENDOCRINOL BRANCH, NIH, BETHESDA, MD 20892 USA. NIH, DEPT RADIOL, BETHESDA, MD 20892 USA. GEORGETOWN UNIV, CHILDRENS MED CTR, DEPT PEDIAT, AS, NORWAY. NR 26 TC 41 Z9 41 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 EI 1945-7197 J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD OCT PY 1996 VL 81 IS 10 BP 3535 EP 3539 DI 10.1210/jc.81.10.3535 PG 5 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VL566 UT WOS:A1996VL56600017 PM 8855797 ER PT J AU Cizza, G Nieman, LK Doppman, JL Passaro, MD Czerwiec, FS Chrousos, GP Cutler, GB AF Cizza, G Nieman, LK Doppman, JL Passaro, MD Czerwiec, FS Chrousos, GP Cutler, GB TI Factitious Cushing syndrome SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID DIFFERENTIAL-DIAGNOSIS; STIMULATION TEST; DISEASE; DEXAMETHASONE; TIME AB There have been few reports of factitious Cushing syndrome. To characterize the clinical and laboratory features leading to this unusual diagnosis, we describe 6 patients (5 women, 1 man), ages 31-44, identified retrospectively among 860 patients evaluated for hypercortisolism at the National Institutes of Health Clinical Center. All six patients had multiple surgeries unrelated to Cushing syndrome and a history of depression or anxiety. Four patients had close contact with the medical profession, three a history of drug abuse, and three had undergone previous treatment for Cushing syndrome. The physical features of Cushing syndrome were variable and not helpful in the differential diagnosis with endogenous Cushing syndrome. Four patients had striking variability in urine-free cortisol (UFC) and 17-hydroxysteroid (17-OHCS) values from low to high. Adrenal computed tomography, performed in two patients, showed small adrenal glands (n = 1) or a left-sided mass (n = 1), and adrenal magnetic resonance imaging, performed in one patient, showed atrophic glands. Pituitary magnetic resonance imaging, carried out in four patients, was either normal (n = 1) or exhibited questionable signs of microadenoma (n = 3). Determination of synthetic glucocorticoids by high pressure liquid chromatography (HPLC) was positive in the four patients in whom it was performed. Factitious Cushing syndrome is a difficult diagnosis. To conserve time and resources, high pressure liquid chromatography analysis of urine steroids, the most definitive test for the factitious disorder, should be performed whenever there is clinical suspicion of glucocorticoid abuse. C1 NIMH, CLIN NEUROENDOCRINOL BRANCH, BETHESDA, MD 20892 USA. NIH, WARREN G MAGNUSON CLIN CTR, DEPT DIAGNOST RADIOL, BETHESDA, MD 20892 USA. RP Cizza, G (reprint author), NICHHD, DEV ENDOCRINOL BRANCH,NIH,NLDG 10,RM 10N-262, 10 CTR DR, BETHESDA, MD 20892 USA. NR 19 TC 34 Z9 34 U1 0 U2 2 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0021-972X EI 1945-7197 J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD OCT PY 1996 VL 81 IS 10 BP 3573 EP 3577 DI 10.1210/jc.81.10.3573 PG 5 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VL566 UT WOS:A1996VL56600023 PM 8855803 ER PT J AU Muesing, RA Forman, MR Graftbard, BI Beecher, GR Lanza, E McAdam, PA Campbell, WS Olson, BR AF Muesing, RA Forman, MR Graftbard, BI Beecher, GR Lanza, E McAdam, PA Campbell, WS Olson, BR TI Cyclic changes in lipoproprotein and apolipoprotein levels during the menstrual cycle in healthy premenopausal women on a controlled diet SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID HIGH-DENSITY-LIPOPROTEIN; PLASMA-LIPIDS; ORAL-CONTRACEPTIVES; CHOLESTEROL; PHASE; SUBCLASSES; ESTRADIOL; FRACTIONS; ESTROGEN; LIPASE AB Lipoprotein, apolipoprotein (ape), and hormone levels were measured in 12 healthy women over three consecutive menstrual cycles; one free-living and two under controlled dietary conditions. Serum hormone levels were measured to identify menstrual cycle phases (menses, early follicular, late follicular, and midluteal). After stabilization for one cycle on the controlled diet, ANOVA modeling of the second controlled-diet cycle revealed that low-density lipoprotein (LDL) cholesterol levels in the midluteal phase were significantly lower (by 7%) than in the early follicular phase. High-density lipoprotein (HDL) cholesterol levels during the late follicular phase were higher (by 6%) than menses levels. Differences in the HDL-cholesterol and apoA-I fluctuations resulted in a higher proportion of HDL-cholesterol to apoA-I during the late follicular phase than that during the menses phase. The ratios of LDL cholesterol/HDL cholesterol and apoB/apoA-I in the early follicular phase were greater by 5.6% and 6.0%, respectively, than those in the midluteal phase. Fluctuations in total cholesterol, triglyceride, apoA-I, and apoB did not reach significance. Thus, the cyclic fluctuations of LDL and HDL cholesterol need to be considered in the screening and medical monitoring of women with borderline lipoprotein levels, as well as in the design and the interpretation of results of studies involving premenopausal women. C1 NCI, NIH, CANC PREVENT & CONTROL BRANCH, BETHESDA, MD 20892 USA. GEORGE WASHINGTON UNIV, MED CTR, DEPT MED, LIPID RES CLIN, WASHINGTON, DC 20037 USA. USDA, HUMAN NUTR RES CTR, BELTSVILLE, MD 20705 USA. MOYNAHAN MED CTR, WATERBURY, CT 06702 USA. NR 32 TC 43 Z9 45 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 OCT PY 1996 VL 81 IS 10 BP 3599 EP 3603 DI 10.1210/jc.81.10.3599 PG 5 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VL566 UT WOS:A1996VL56600028 PM 8855808 ER PT J AU Stratakis, CA Jenkins, RB Pras, E Mitsiadis, CS Raff, SB Stalboerger, PG Tsigos, C Carney, JA Chrousos, GP AF Stratakis, CA Jenkins, RB Pras, E Mitsiadis, CS Raff, SB Stalboerger, PG Tsigos, C Carney, JA Chrousos, GP TI Cytogenetic and microsatellite alterations in tumors from patients with the syndrome of myxomas, spotty skin pigmentation, and endocrine overactivity (Carney complex) SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID PSAMMOMATOUS MELANOTIC SCHWANNOMA; FAMILIAL CARDIAC MYXOMA; SPECIAL ASSOCIATIONS; MALIGNANT-MELANOMA; CUTANEOUS MYXOMAS; COLORECTAL-CANCER; FREQUENT LOSS; HETEROZYGOSITY; MULTIPLE; GENE AB Carney complex (CC) is a familial multiple neoplasia and lentiginosis syndrome, transmitted in an autosomal dominant manner. It is the only familial form of cardiac and skin myxomas known and in eludes endocrine neoplasms causing Cushing's syndrome [primary pigmented nodular adrenocortical disease (PPNAD)] and acromegaly (GH-producing adenoma). The molecular defect leading to CC remains unknown, but was recently mapped to chromosome 2p16 by linkage analysis. This region has exhibited cytogenetic aberrations in atrial myxomas from patients with CC and harbors the hMSH2 and hMSH6 genes, which are involved in the preservation of microsatellite length stability of replicating human cells. In the present study, we examined 15 tumor and normal tissue specimens from 13 patients with CC [GH-producing adenoma (n = 1), adrenal tumors (PPNAD: n = 8), thyroid cancer (n = 1), normal adrenal gland (n = 1)] and 4 cultured cell lines [heart myxoma (n = 3) and eyelid myxoma(n = 1)]. Chromosome analysis was obtained by standard cytogenetic techniques. One of the myxoma cell lines and 3 PPNAD specimens contained multiple telomeric associations (tas). The normal adrenocortical tissue from a patient with PPNAD contained no apparent chromosomal anomalies, whereas the neighboring PPNAD tissue demonstrated tas. DNA was extracted from peripheral blood, tumor cell lines, and frozen or paraffin-embedded tissues and subjected to PCR amplification with primers from 64 microsatellite locations covering chromosomes 1 and 3-22 and 14 loci covering chromosome 2. The alterations detected were loss and gain of heterozygosity (LOH and GOH; 49% and 26%, respectively), deletions of both alleles (DEL; 10%), and microsatellite length instability (15%). GOH and LOH were the most frequent changes, with telomeric markers significantly overrepresented (P < 0.05). Chromosomes 6, 11, 22, 10, and 19 demonstrated mostly LOH, GOH, or DEL in over 40% of the informative loci tested (73%, 59%, 47%, 46%, and 44%, respectively), whereas markers on chromosome 2 showed only microsatellite length instability (10%). The degree of genomic instability and its type were independent of tumor type (P > 0.1). We conclude that tumors and tumor cell lines from patients with CC demonstrate significant genomic, but not microsatellite length, instability. Thus, the CC gene(s) on chromosome 2p16 is different from the hMSH2 and hMSH6 genes and has dominant, rather than recessive, tumorigenic function. This gene(s) appears to be involved in the regulation of genomic stability of dividing cells, in particular the structure of telomeres in replicating chromosomes and/or the function of the mitotic apparatus. C1 MAYO CLIN, DEPT LAB MED & PATHOL, ROCHESTER, MN 55905 USA. UNIV CONNECTICUT, ENDOCRINOL SECT, NEW BRITAIN GEN HOSP, STORRS, CT 06050 USA. NIAMSD, GENET SECT, ARTHRIT & RHEUMATISM BRANCH, BETHESDA, MD 20892 USA. RP Stratakis, CA (reprint author), NICHHD, NIH,SECT PEDIAT ENDOCRINOL,DEV ENDOCRINOL BRANCH, BLDG 10,ROOM 10N 262, 10 CTR DR, BETHESDA, MD 20892 USA. NR 54 TC 71 Z9 73 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 EI 1945-7197 J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD OCT PY 1996 VL 81 IS 10 BP 3607 EP 3614 DI 10.1210/jc.81.10.3607 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VL566 UT WOS:A1996VL56600030 PM 8855810 ER PT J AU Schuppert, F Taniguchi, S Schroder, S AF Schuppert, F Taniguchi, S Schroder, S TI In vivo and in vitro evidence for iodide regulation of major histocompatibility complex class I and class II expression in Graves' disease SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID AUTOIMMUNE THYROID-DISEASE; SYSTEMIC LUPUS-ERYTHEMATOSUS; HLA-DR EXPRESSION; T-CELL CLONES; MHC CLASS-II; GENE-EXPRESSION; THYROTROPIN RECEPTOR; ANTIGEN EXPRESSION; INTERFERON; INDUCTION AB Increases in thyroid cell major histocompatibility complex (MHC) class I and class II expression have been suggested to be an important factor in the development or perpetuation of Graves' disease. It is hypothesized that elevations result in abnormal presentation of thyroid antigens to immune cells, and that iodide and/or methimazole (MMI) are effective therapeutic agents because, at least in part, of their suppression of MHC expression. In this report, we show that Graves' patients pretreated with iodide only 4 days before surgery have lower levels of MHC class I and class II RNA levels in their thyroid tissue than do patients with no iodide pretreatment (P < 0.001 and 0.03, respectively). Because patients in both groups are treated with MMI and because the change is independent of the amounts of MMI used to treat patients, the class I and class II changes cannot be ascribed to MMI. The iodide action to decrease MHC class I and class II RNA levels was duplicated using cultured human thyroid cells in vitro; the iodide effect was dependent on the iodide concentration, was not duplicated by chloride, was not associated with an alteration in cAMP levels or with a change in thyrotropin receptor RNA levels, and was evident in gamma-interferon-treated cells. The data suggest, therefore, that the therapeutic action of iodide in Graves' patients is associated with decreased MHC gene expression, that this action is a direct effect of high concentrations of iodide on the thyroid cells, and that altered MHC gene expression in the target tissue may well be associated with the development or perpetuation of Graves' disease. C1 NIDDK, NIH, METAB DIS BRANCH, BETHESDA, MD USA. INST IMMUNOL PATHOL & MOL BIOL, HAMBURG, GERMANY. UNIV HALLE WITTENBERG, DEPT SURG, HALLE, GERMANY. RP Schuppert, F (reprint author), HANNOVER MED SCH, DEPT CLIN ENDOCRINOL, KONSTANTY GUTSCHOW STR 8, D-30625 HANNOVER, GERMANY. NR 33 TC 18 Z9 20 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 OCT PY 1996 VL 81 IS 10 BP 3622 EP 3628 DI 10.1210/jc.81.10.3622 PG 7 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VL566 UT WOS:A1996VL56600033 PM 8855812 ER PT J AU Reincke, M Wachenfeld, C Mora, P Thumser, A JaurschHancke, C Abdelhamid, S Chrousos, GP Allolio, B AF Reincke, M Wachenfeld, C Mora, P Thumser, A JaurschHancke, C Abdelhamid, S Chrousos, GP Allolio, B TI p53 mutations in adrenal tumors: Caucasian patients do not show the exon 4 ''hot spot'' found in Taiwan SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID SUPPRESSOR GENE; POLYMERASE; NEOPLASMS; CANCERS; DNA AB Mutations in the p53 tumor suppressor gene are frequently present in human cancers but have rarely been described in benign tumors. We previously reported mutations in the ''hot spots'' between exons 5-8 of the p53 gene in adrenocortical carcinomas but not in adenomas. Recently, a previously unknown hot spot in exon 4 of the p53 gene was described in adrenal adenomas and pheochromocytomas of Taiwanese patients. We, therefore, investigated whether these mutations are also present in Caucasian patients from the U.S. and Europe. We analyzed tumor tissue of 12 aldosterone-producing adenomas, 7 cortisol-producing adenomas, and 6 pheochromocytomas. Overexpression of the p53 protein was investigated by immunohistochemistry. Point mutations within exon 4 were identified by polymerase chain reaction (PCR) amplification and direct sequencing of the PCR product. The pYNZ22 microsatellite located on chromosome 17p, close to the p53 gene, was used to screen for allelic loss (LOH) of the p53 gene. Overexpression of p53 was not identified in any of the adenomas and pheochromocytomas. Point mutations within exon 4 were found in 0/25 tumors. LOH was present in 1/13 informative adenomas and 0/2 informative pheochromocytomas. We conclude that p53 mutations do not play a major role in the tumorigenesis of adrenal adenomas and pheochromocytomas of Caucasian patients. Thus, ethnic and environmental factors may be responsible for the mutational spectrum found in Taiwanese patients. C1 NICHHD, NIH, DEV ENDOCRINOL BRANCH, BETHESDA, MD 20892 USA. RP Reincke, M (reprint author), UNIV WURZBURG, MED KLIN, DTSCH KLIN DIAG, JOSEF SCHNEIDER STR 2, D-97080 WURZBURG, GERMANY. NR 20 TC 42 Z9 43 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 EI 1945-7197 J9 J CLIN ENDOCR METAB JI J. Clin. Endocrinol. Metab. PD OCT PY 1996 VL 81 IS 10 BP 3636 EP 3638 DI 10.1210/jc.81.10.3636 PG 3 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VL566 UT WOS:A1996VL56600035 PM 8855814 ER PT J AU Ferrucci, L Guralnik, JM Salive, ME Fried, LP BandeenRoche, K Brock, DB Simonsick, EM Corti, MC Zeger, SL AF Ferrucci, L Guralnik, JM Salive, ME Fried, LP BandeenRoche, K Brock, DB Simonsick, EM Corti, MC Zeger, SL TI Effect of age and severity of disability on short-term variation in walking speed: The women's health and aging study SO JOURNAL OF CLINICAL EPIDEMIOLOGY LA English DT Article DE walking; performance; aging; physical function; disability; autocorrelation ID LOWER-EXTREMITY FUNCTION; PHYSICAL PERFORMANCE; BUTTON TEST; RHEUMATOID-ARTHRITIS; STANDING BALANCE; GRIP STRENGTH; HEART-FAILURE; OLDER ADULTS; MORTALITY; DISEASE AB Standardized objective measures of human performance have been introduced in clinical and epidemiologic studies of older populations. Reliability of these measures has usually been estimated by comparing two measures obtained in the same person. However, no information is available on variability of multiple measures collected serially over short time intervals. This study uses data from the Weekly Disability Study, a component of the Women's Health and Aging Study, to describe fluctuations in physical performance over multiple, consecutive time intervals. Walking speed was measured weekly over a 6 month period in 99 older women affected by mild to severe disability. Overall, 2120 observations were explored using techniques developed for the analysis of repeated measures. Results showed that the correlations between observations in the same person were inversely related to their separation in time. The decay in the autocorrelation function was steeper in the least disabled. However, even with 20-week separations in assessments, correlations remained above 0.6 in all age and severity of disability subgroups. Changes over time in performance differed somewhat between disability subgroups, but the relative performance across subgroups remained stable over the entire course of the study. A clear learning effect was found only in those in the middle disability subgroup. Results support the utilization of repeated measures of physical performance in research that evaluates older persons over time. C1 NATL RES INST,DEPT GERIATR,FLORENCE,ITALY. JOHNS HOPKINS MED INST,SCH MED,DEPT MED,BALTIMORE,MD 21205. JOHNS HOPKINS MED INST,SCH MED,DEPT EPIDEMIOL,BALTIMORE,MD 21205. JOHNS HOPKINS MED INST,SCH HYG & PUBL HLTH,DEPT BIOSTAT,BALTIMORE,MD 21205. RP Ferrucci, L (reprint author), NIA,EPIDEMIOL DEMOG & BIOMETRY PROGRAM,7201 WISCONSIN AVE,GATEWAY BLDG,SUITE 3C-309,BETHESDA,MD 20892, USA. NR 38 TC 31 Z9 31 U1 4 U2 5 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, ENGLAND OX5 1GB SN 0895-4356 J9 J CLIN EPIDEMIOL JI J. Clin. Epidemiol. PD OCT PY 1996 VL 49 IS 10 BP 1089 EP 1096 DI 10.1016/0895-4356(96)00231-4 PG 8 WC Health Care Sciences & Services; Public, Environmental & Occupational Health SC Health Care Sciences & Services; Public, Environmental & Occupational Health GA VK718 UT WOS:A1996VK71800004 PM 8826987 ER PT J AU Lazarova, Z Yee, C Darling, T Briggaman, RA Yancey, KB AF Lazarova, Z Yee, C Darling, T Briggaman, RA Yancey, KB TI Passive transfer of anti-laminin 5 antibodies induces subepidermal blisters in neonatal mice SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article DE autoimmunity; laminin; animal models ID JUNCTIONAL EPIDERMOLYSIS-BULLOSA; EPIDERMAL BASEMENT-MEMBRANE; MONOCLONAL-ANTIBODY; HERPES GESTATIONIS; GENE LAMC2; HG-FACTOR; KERATINOCYTES; ADHESION; AUTOANTIBODIES; EPILIGRIN AB Patients with a recently identified subepithelial blistering disease have IgG anti-laminin 5 autoantibodies. To determine if such antibodies can be pathogenic in vivo, we developed and characterized rabbit anti-laminin 5 IgG, and passively transferred these antibodies to neonatal mice. Immune rabbit IgG specifically bound human and murine epidermal basement membranes, immunoblotted and immunoprecipitated all laminin 5 subunits from extracts of human and murine keratinocytes, and showed no reactivity to other keratinocyte proteins or epithelial basement membranes that do not contain laminin 5. Mice (n=29) receiving purified anti-laminin 5 IgG developed, in a dose-related fashion, circulating anti-laminin 5 antibodies, deposits of rabbit IgG and murine C3 in epidermal basement membranes, and subepidermal blisters of skin and mucous membranes. No alterations developed in controls (n=14) receiving identical amounts of normal rabbit IgG. Passive transfer of antilaminin 5 (but not control) IgG to neonatal C5- (n=3) or mast cell-deficient (n=3) mice produced subepidermal blisters with the same clinical, histologic, and immunopathologic features as those documented in BALB/c mice. These studies establish an animal model of a human blistering disease that can be used to define disease mechanisms and treatment modalities. C1 NCI,DERMATOL BRANCH,NIH,BETHESDA,MD 20892. UNIV N CAROLINA,DEPT DERMATOL,CHAPEL HILL,NC 27514. OI Darling, Thomas/0000-0002-5161-1974 NR 33 TC 115 Z9 116 U1 0 U2 2 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 222 E 70TH STREET, NEW YORK, NY 10021 SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD OCT 1 PY 1996 VL 98 IS 7 BP 1509 EP 1518 DI 10.1172/JCI118942 PG 10 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA VL112 UT WOS:A1996VL11200004 PM 8833897 ER PT J AU Elliott, EA McFarland, HI Nye, SH Cofiell, R Wilson, TM Wilkins, JA Squinto, SP Matis, LA Mueller, JP AF Elliott, EA McFarland, HI Nye, SH Cofiell, R Wilson, TM Wilkins, JA Squinto, SP Matis, LA Mueller, JP TI Treatment of experimental encephalomyelitis with a novel chimeric fusion protein of myelin basic protein and proteolipid protein SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article DE autoimmunity; apoptosis; T cells; tolerance; immunomodulators ID EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS; EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS; MULTIPLE-SCLEROSIS PATIENTS; T-CELL CLONES; LYMPH-NODE CELLS; ADOPTIVE TRANSFER; ENCEPHALITOGENIC EPITOPE; DEMYELINATING DISEASES; HEALTHY-INDIVIDUALS; CEREBROSPINAL-FLUID AB It has been shown that peripheral T cell tolerance can be induced by systemic antigen administration. We have been interested in using this phenomenon to develop antigen-specific immunotherapies for T cell-mediated autoimmune diseases. In patients with the demyelinating disease multiple sclerosis (MS), multiple potentially autoantigenic epitopes have been identified on the two major proteins of the myelin sheath, myelin basic protein (MBP) and proteolipid protein (PLP). To generate a tolerogenic protein for the therapy of patients with MS, we have produced a protein fusion between the 21.5-kD isoform of MBP (MBP21.5) and a genetically engineered form of PLP (Delta PLP4). In this report, we describe the effects of treatment with this agent (MP4) on clinical disease in a murine model of demyelinating disease, experimental autoimmune encephalomyetitis (EAE). Treatment of SJL/J mice with MP4 after induction of EAE either by active immunization or by adoptive transfer of activated T cells completely prevented subsequent clinical paralysis. Importantly, the administration of MP4 completely suppressed the development of EAE initiated by the cotransfer of both MBP- and PLP-activated T cells. Prevention of clinical disease after the intravenous injection of MP4 was paralleled by the formation of long-lived functional peptide-MHC complexes in vivo, as well as by a significant reduction in both MBP- and PLP-specific T cell proliferative responses. Mice treated with MP4 were resistant to disease when rechallenged with an encephalitogenic PLP peptide emulsified in CFA, indicating that MP4 administration had a prolonged effect in vivo. Administration of MP4 was also found to markedly ameliorate the course of established clinical disease. Finally, MP4 therapy was equally efficacious in mice defective in Fas expression, These results support the conclusion that MP4 protein is highly effective in suppressing disease caused by multiple neuroantigen epitopes in experimentally induced demyelinating disease. C1 ALEX PHARMACEUT INC,DEPT IMMUNOBIOL,NEW HAVEN,CT 06511. NIAID,IMMUNOL LAB,NIH,BETHESDA,MD 20892. ALEX PHARMACEUT INC,DEPT MOL DEV,NEW HAVEN,CT 06511. ALEX PHARMACEUT INC,DEPT PROC DEV,NEW HAVEN,CT 06511. RI McFarland, Hugh/K-1503-2016 OI McFarland, Hugh/0000-0002-3322-038X NR 68 TC 71 Z9 71 U1 0 U2 0 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 222 E 70TH STREET, NEW YORK, NY 10021 SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD OCT 1 PY 1996 VL 98 IS 7 BP 1602 EP 1612 DI 10.1172/JCI118954 PG 11 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA VL112 UT WOS:A1996VL11200016 PM 8833909 ER PT J AU Schwan, TG Schrumpf, ME Hinnebusch, BJ Anderson, DE Konkel, ME AF Schwan, TG Schrumpf, ME Hinnebusch, BJ Anderson, DE Konkel, ME TI GlpQ: An antigen for serological discrimination between relapsing fever and Lyme borreliosis SO JOURNAL OF CLINICAL MICROBIOLOGY LA English DT Article ID LINKED IMMUNOSORBENT ASSAYS; D-BINDING PROTEIN; ESCHERICHIA-COLI; HAEMOPHILUS-INFLUENZAE; SURFACE PROTEIN; GLYCEROPHOSPHODIESTER PHOSPHODIESTERASE; NUCLEOTIDE-SEQUENCE; ANTIBODY-RESPONSE; CHAIN-REACTION; DNA INVITRO AB Tick-borne relapsing fever is caused by numerous Borrelia species maintained in nature by Ornithodoros tick-mammal cycles. Serological confirmation is based on either an immunofluorescence assay or an enzyme-linked immunosorbent assay using whole cells or sonicated Borrelia hermsii as the antigen, However, antigenic variability of this bacterium's outer surface proteins and antigens shared with the Lyme disease spirochete (B. burgdorferi), may cause both false-negative and false-positive results when testing sera of patients suspected to have either relapsing fever or Lyme disease, To develop a specific serological test for relapsing fever, we created a genomic DNA library of B. hermsii, screened transformed Escherichia coli cells for immunoreactivity with high-titered (greater than or equal to 1:2,048) human anti-B. hermsii antiserum, and selected an immunoreactive clone (pSPR75) expressing a 39-kDa protein, DNA sequencing, subcloning, and serum adsorption experiments identified the immunoreactive protein as a homolog of GlpQ, a glycerophosphodiester phosphodiesterase identified previously in E. coli, Haemophilus influenzae, and Bacillus subtilis. Serum samples from humans and mice infected with B. hermsii or other species of relapsing fever spirochetes contained antibodies recognizing GlpQ, whereas serum samples from Lyme disease and syphilis patients were nonreactive. Serologic tests based on this antigen will identify people exposed previously to relapsing fever spirochetes and help clarify the distribution of relapsing fever and Lyme disease in situations in which the occurrence of their causative agents is uncertain. C1 SACRED HEART MED CTR,SPOKANE,WA 99220. WASHINGTON STATE UNIV,DEPT MICROBIOL,PULLMAN,WA 99164. RP Schwan, TG (reprint author), NIAID,ROCKY MT LABS,NIH,MICROBIAL STRUCT & FUNCT LAB,HAMILTON,MT 59840, USA. NR 54 TC 82 Z9 87 U1 0 U2 2 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0095-1137 J9 J CLIN MICROBIOL JI J. Clin. Microbiol. PD OCT PY 1996 VL 34 IS 10 BP 2483 EP 2492 PG 10 WC Microbiology SC Microbiology GA VK787 UT WOS:A1996VK78700029 PM 8880505 ER PT J AU Kuttesch, JF Wexler, LH Marcus, RB Fairclough, D WeaverMcClure, L White, M Mao, L Delaney, TF Pratt, CB Horowitz, ME Kun, LE AF Kuttesch, JF Wexler, LH Marcus, RB Fairclough, D WeaverMcClure, L White, M Mao, L Delaney, TF Pratt, CB Horowitz, ME Kun, LE TI Second malignancies after Ewing's sarcoma: Radiation dose-dependency of secondary sarcomas SO JOURNAL OF CLINICAL ONCOLOGY LA English DT Article ID COMBINED MODALITY THERAPY; CHILDHOOD-CANCER; MULTIMODAL THERAPY; BONE SARCOMAS; LOCAL-CONTROL; YOUNG-ADULTS; NEOPLASMS; CHILDREN; RISK; LEUKEMIA AB Background: An excess risk of second malignancies has been reported in survivors of Ewing's sarcoma. We examined a multiinstitutional data base to reevaluate the risk among survivors of Ewing's sarcoma and to identify possible causal factors. Methods: Information was derived from a data base that included 266 survivors of Ewing's sarcoma. Cumulative incidence rates of second malignancies were calculated. Contributions of clinical features, type and dose of chemotherapy, and cumulative radiation dose to the risk of second malignancies were evaluated. Results: After a median follow-up duration of 9.5 years (range, 3.0 to 30), 16 patients have developed second malignancies, which included 10 sarcomas (five osteosarcomas, three fibrosarcomas, and two malignant fibrous histiocytomas) and six other malignancies (acute myeloblastic leukemia, acute lymphoblastic leukemia, meningioma, bronchioalveolar carcinoma, basal cell carcinoma, and carcinoma-in-situ of the cervix), The median latency to the diagnosis of the second malignancy was 7.6 years (range, 3.5 to 25.7). The estimated cumulative incidence rates at 20 years for any second malignancy and for secondary sarcoma were 9.2% (SD = 2.7%) and 6.5% (SD = 2.4%), respectively. The cumulative incidence rate of secondary sarcoma was radiation dose-dependent (P = .002). No secondary sarcomas developed among patients who had received less than 48 Gy, while the absolute risk of secondary sarcoma was 130 cases per 10,000 person-years of observation among patients who had received greater than or equal to 60 Gy. Conclusion: The overall risk of second malignancies after Ewing's sarcomas is similar to that associated with treatment for other childhood cancers. The radiation dose-dependency of secondary sarcomas justifies modification in therapy to reduce radiation doses. (C) 1996 by American Society of Clinical Oncology. C1 UNIV TENNESSEE, COLL MED, DEPT PEDIAT, MEMPHIS, TN USA. UNIV TENNESSEE, COLL MED, DEPT RADIAT ONCOL, MEMPHIS, TN USA. UNIV FLORIDA, SCH MED, DEPT RADIAT ONCOL, GAINESVILLE, FL USA. NCI, PEDIAT BRANCH, BETHESDA, MD 20892 USA. ST JUDE CHILDRENS RES HOSP, DEPT HEMATOL & ONCOL, MEMPHIS, TN 38105 USA. ST JUDE CHILDRENS RES HOSP, DEPT BIOSTAT, MEMPHIS, TN 38105 USA. ST JUDE CHILDRENS RES HOSP, DEPT RADIAT ONCOL, MEMPHIS, TN 38105 USA. RI Mao, Li/C-7570-2011 OI Mao, Li/0000-0001-7263-3358 FU NCI NIH HHS [CA 23099, P30 CA-21765] NR 40 TC 176 Z9 185 U1 0 U2 3 PU AMER SOC CLINICAL ONCOLOGY PI ALEXANDRIA PA 2318 MILL ROAD, STE 800, ALEXANDRIA, VA 22314 USA SN 0732-183X J9 J CLIN ONCOL JI J. Clin. Oncol. PD OCT PY 1996 VL 14 IS 10 BP 2818 EP 2825 PG 8 WC Oncology SC Oncology GA VN185 UT WOS:A1996VN18500026 PM 8874344 ER PT J AU Bast, RC Bates, S Bredt, AB Desch, CE Fritsche, H Fues, L Hayes, DF Kemeny, NE Kragen, M Jessup, J Locker, GY Macdonald, JS Mennel, RG Norton, L Ravdin, P Smith, TJ Taube, S Winn, RJ AF Bast, RC Bates, S Bredt, AB Desch, CE Fritsche, H Fues, L Hayes, DF Kemeny, NE Kragen, M Jessup, J Locker, GY Macdonald, JS Mennel, RG Norton, L Ravdin, P Smith, TJ Taube, S Winn, RJ TI Clinical practice guidelines for the use of tumor markers in breast and colorectal cancer SO JOURNAL OF CLINICAL ONCOLOGY LA English DT Review ID CARCINOEMBRYONIC ANTIGEN CEA; FLOW-CYTOMETRIC ANALYSIS; S-PHASE FRACTION; ESTROGEN-RECEPTOR ANALYSIS; PARAFFIN-EMBEDDED TISSUE; SERUM CA-125 LEVELS; LONG-TERM SURVIVAL; C-ERBB-2 ONCOPROTEIN EXPRESSION; INDEPENDENT PROGNOSTIC VALUE; SURGICAL ADJUVANT BREAST AB Purpose: The primary objective was to determine clinical practice guidelines for the use of tumor marker tests in the prevention, screening, treatment, and surveillance of breast and colorectal cancers. These guidelines are intended for use in the care of patients outside of clinical trials. Methods: Six tumor markers for colorectal cancer: and seven for breast cancer were considered. The could be recommended or not for routine use or for special circumstances. In general, the significant health outcomes identified for use in making clinical practice guidelines (overall survival, disease-free survival, quality of life, lesser toxicity, and cost-effectiveness) were used when data were available. Expert consensus was used to inform the recommendations for use of tumor markers when published evidence was insufficient. A computerized literature search was performed using Medline. In addition to reports collected by individual panel members, all articles published in the English-speaking literature from January 1989 to April 1994 were collected for review and distributed to all members of the Panel. Values for use, utility, and levels of evidence were assigned by the expert reviewers and approved by the Panel. Tumor markers were assigned benefit if they had prognostic or predictive value that would lead to the favorable outcomes listed above. Harms considered were inappropriate disease management, and excess cost without definable benefit. Costs were considered but were never the sole determinant of a recommendation. Results and Conclusion: For colorectal cancer, it is recommended that carcinoembryonic antigen (CEA) levels be measured preoperatively if it would change surgical management. It is recommended that CEA levels be monitored every 2 to 3 months for greater than or equal to 2 years, if resection of liver metastasis would be clinically indicated. The data are insufficient to recommend the routine use of lipid-associated sialic acid (LASA), CA 19-9, DNA index, DNA flow cytometric proliferation analysis, 953 tumor suppressor gene, and ras oncogene. For breast cancer, estrogen receptor and progesterone receptor are recommended to be measured on every primary specimen, but on subsequent specimens only if it would lead to a change in management. The data Fire insufficient to recommend the routine use of DNA index, DNA flow cytometric proliferation analysis, CA 15-3, CEA, c-erbs-2, 953 or cathepsin-D. In the absence of readily measurable disease, CA 15-3 and CEA levels can be used to document treatment failure. New markers and new evidence will be evaluated by annual update of these guidelines. (C) 1996 by American Society of Clinical Oncology. C1 UNIV TEXAS,MD ANDERSON CANC CTR,HOUSTON,TX. NIH,BETHESDA,MD 20892. SO CALIF KAISER PERMANENTE,NORTHRIDGE,CA. VIRGINIA COMMONWEALTH UNIV,MED COLL VIRGINIA,MASSEY CANC CTR,RICHMOND,VA 23298. DANA FARBER CANC INST,BOSTON,MA 02115. MEM SLOAN KETTERING CANC CTR,NEW YORK,NY 10021. NEW ENGLAND DEACONESS HOSP,BOSTON,MA 02215. NORTHWESTERN UNIV,EVANSTON HOSP,EVANSTON,IL 60201. TEMPLE UNIV,CTR CANC,PHILADELPHIA,PA 19122. UNIV TEXAS,HLTH SCI CTR,SAN ANTONIO,TX. NCI,CANC DIAG BRANCH,DIV CANC BIOL DIAG & CTR,ROCKVILLE,MD. RP Bast, RC (reprint author), AMER SOC CLIN ONCOL,HLTH SERV RES,225 REINEKERS LANE,SUITE 650,ALEXANDRIA,VA 22314, USA. RI Bast, Robert/E-6585-2011 OI Bast, Robert/0000-0003-4621-8462 NR 380 TC 392 Z9 395 U1 4 U2 17 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0732-183X J9 J CLIN ONCOL JI J. Clin. Oncol. PD OCT PY 1996 VL 14 IS 10 BP 2843 EP 2877 PG 35 WC Oncology SC Oncology GA VN185 UT WOS:A1996VN18500029 ER PT J AU Klausner, RD AF Klausner, RD TI The future of cancer research and the role of the National Cancer Institute SO JOURNAL OF CLINICAL ONCOLOGY LA English DT Article RP Klausner, RD (reprint author), NCI,BLDG 31,ROOM 1A48,9000 ROCKVILLE PIKE,BETHESDA,MD 20892, USA. NR 0 TC 3 Z9 3 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0732-183X J9 J CLIN ONCOL JI J. Clin. Oncol. PD OCT PY 1996 VL 14 IS 10 BP 2878 EP 2883 PG 6 WC Oncology SC Oncology GA VN185 UT WOS:A1996VN18500030 PM 8874348 ER PT J AU Murphy, WJ Muroi, M Zhang, CX Suzuki, T Russell, SW AF Murphy, WJ Muroi, M Zhang, CX Suzuki, T Russell, SW TI Both basal and enhancer kappa B elements are required for full induction of the mouse inducible nitric oxide synthase gene SO JOURNAL OF ENDOTOXIN RESEARCH LA English DT Article ID TRANSCRIPTION FACTOR; INTERFERON-GAMMA; CELL-LINE; BACTERIAL LIPOPOLYSACCHARIDE; INTERLEUKIN-6 GENE; NUCLEAR FACTOR; T-CELLS; MACROPHAGES; ACTIVATION; EXPRESSION AB The transcriptional regulatory region of the mouse inducible nitric oxide synthase (iNOS) gene has two kappa B elements, one enhancer-linked (kappa BII) and the other (kappa BI) proximal to its core promoter. Mutation of kappa BII substantially reduced the extent to which the iNOS promoter could be induced by IFS and interfered with augmented responsiveness of the promoter to LPS+IFN-gamma. Mutation of kappa BI had a quantitatively less dramatic negative effect on LPS responsiveness and this construct still showed augmented responsiveness to LPS+IFN-gamma. When both kappa B elements were mutated, inducibility by LPS and, in particular, by LPS+IFN-gamma was paradoxically restored, compared with the mutated kappa BII alone, suggesting cooperative interactions among the transcription factors that trans-activate the iNOS gene. In vivo footprint analysis showed that both kappa B elements were bound by protein complexes when macrophages were stimulated with LPS +/- IFN-gamma. Furthermore, kappa BI was bound even in untreated cells, suggesting that kappa B binding proteins might also have a negative influence on expression of the gene. Both kappa BI and kappa BII were bound by NF-kappa B/Rel proteins found in nuclear extracts prepared from macrophages treated with LPS + IFN-gamma, although the specificity of binding to each element was different. Our results show that, while NF-kappa B/Rel proteins are required for maximal expression of the iNOS gene, alone they are not alone sufficient. Furthermore, the results reported here show that the augmentative effect of IFN-gamma on the LPS-induced expression of the INOS gene is not mediated through increased activation of NF-kappa B/Rel. C1 UNIV KANSAS,MED CTR,DEPT PATHOL & LAB MED,KANSAS CITY,KS 66160. UNIV KANSAS,MED CTR,DEPT MICROBIOL MOL GENET & IMMUNOL,KANSAS CITY,KS 66160. NCI,FREDERICK CANC RES & DEV CTR,ABL BASIC RES PROGRAM,FREDERICK,MD. RP Murphy, WJ (reprint author), KANSAS CANC INST,WILKINSON LAB,1008 WAHL HALL W,3901 RAINBOW BLVD,KANSAS CITY,KS 66160, USA. NR 51 TC 29 Z9 30 U1 0 U2 4 PU CHURCHILL LIVINGSTONE PI EDINBURGH PA JOURNAL PRODUCTION DEPT, ROBERT STEVENSON HOUSE, 1-3 BAXTERS PLACE, LEITH WALK, EDINBURGH EH1 3AF, MIDLOTHIAN, SCOTLAND SN 0968-0519 J9 J ENDOTOXIN RES JI J. Endoxtin Res. PD OCT PY 1996 VL 3 IS 5 BP 381 EP 393 PG 13 WC Biochemistry & Molecular Biology; Immunology; Medicine, Research & Experimental; Microbiology SC Biochemistry & Molecular Biology; Immunology; Research & Experimental Medicine; Microbiology GA WA532 UT WOS:A1996WA53200002 ER PT J AU Plaksin, D Chacko, S McPhie, P Bax, A Padlan, EA Margulies, DH AF Plaksin, D Chacko, S McPhie, P Bax, A Padlan, EA Margulies, DH TI A T cell receptor V alpha domain expressed in bacteria: Does it dimerize in solution? SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Article ID CLASS-I MOLECULE; ESCHERICHIA-COLI; CRYSTAL-STRUCTURE; ANTIGEN RECEPTOR; 3-DIMENSIONAL STRUCTURE; IMMUNOGLOBULIN DOMAIN; MONOCLONAL-ANTIBODY; CIRCULAR-DICHROISM; FAB-FRAGMENTS; PEPTIDE AB To evaluate the potential for dimerization through a particular T cell receptor (TCR) domain, we have cloned the cDNA encoding a TCR V alpha from a hybridoma with specificity for the human immunodeficiency virus (HIV) envelope glycoprotein 120-derived peptide P18-I10 (RGPGRAFVTI) bound to the murine major histocompatibility complex (MHC) class I molecule, H-2D(d). This cDNA was then expressed in a bacterial vector, and protein, as inclusion bodies, was solubilized, refolded, and purified to homogeneity. Yield of the refolded material was from 10 to 50 mg per liter of bacterial culture, the protein was soluble at concentrations as high as 25 mg/ml, and it retained a high level of reactivity with an anti-V alpha 2 monoclonal antibody. This domain was monomeric both by size exclusion gel chromatography and by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Circular dichroism spectra indicated that the folded V alpha domain had secondary structure similar to that of single immunoglobulin or TCR domains, consisting largely of beta sheet. Conditions for crystallization were established, and at least two crystal geometries were observed: hexagonal bipyramids that failed to diffract beyond similar to 6 Angstrom, and orthorhombic crystals that diffracted to 2.5 Angstrom. The dimerization of the V alpha domain was investigated further by solution nuclear magnetic resonance spectroscopy, which indicated that dimeric and monomeric forms of the protein were about equally populated at a concentration of 1 mM. Thus, models of TCR-mediated T cell activation that invoke TCR dimerization must consider that some V alpha domains have little tendency to form homodimers or multimers. C1 NIAID,MOL BIOL SECT,IMMUNOL LAB,NIH,BETHESDA,MD 20892. NIADDKD,MOL BIOL LAB,NIH,BETHESDA,MD 20892. NIADDKD,LAB BIOMOL PHARMACOL,NIH,BETHESDA,MD 20892. NIADDKD,CHEM PHYS LAB,NIH,BETHESDA,MD 20892. RI Margulies, David/H-7089-2013; OI Margulies, David/0000-0001-8530-7375 NR 53 TC 15 Z9 15 U1 0 U2 0 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 222 E 70TH STREET, NEW YORK, NY 10021 SN 0022-1007 J9 J EXP MED JI J. Exp. Med. PD OCT 1 PY 1996 VL 184 IS 4 BP 1251 EP 1258 DI 10.1084/jem.184.4.1251 PG 8 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA VP230 UT WOS:A1996VP23000007 PM 8879196 ER PT J AU Pospisil, R Fitts, MG Mage, RG AF Pospisil, R Fitts, MG Mage, RG TI CD5 is a potential selecting ligand for B cell surface immunoglobulin framework region sequences SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Article ID CLONAL DELETION; LYMPHOCYTE-B; T-CELLS; RABBIT; EXPRESSION; TOLERANCE; ANTIBODY; REPERTOIRE; DIVERSITY; ALLOTYPES AB In rabbits nearly all B lymphocytes express the glycoprotein CD5, in contrast to mice and humans, where only a small proportion of B cells express this molecule (Raman, C., and K.L. Knight. 1992. J. Immunol. 149:3858-3864). CD5(+) B cells appear to develop early in ontogeny and be maintained throughout life by self-renewal. The function of CD5 on B cells is still unknown. We showed earlier that ''positive'' selection occurs during B lymphocyte development in the rabbit appendix. This selection favors B cells expressing surface immunoglobulins with V(H)a2 structures in the first and third framework regions (Pospisil, R., G.O. Young-Cooper, and R.G. Mage. 1995. Proc. Natl. Acad. Sci. USA. 92:6961-6965). Here we report that F(ab')(2) fragments, especially those bearing V(H)a2 framework region determinants, specifically interact with the B cell-surface glycoprotein CD5. This interaction can be inhibited by anti-CD5 antibodies. Furthermore, immobilized F(ab')(2) fragments selectively bind CD5 molecules in appendix cell lysates. Interactions of V-H framework region structures with CD5 may affect maintenance and selective expansion of particular B cells and thus contribute to autostimulatory growth of autoimmune or transformed cells. C1 NIAID,IMMUNOL LAB,NIH,BETHESDA,MD 20892. RI Pospisil, Richard/B-7467-2012 NR 34 TC 66 Z9 66 U1 0 U2 0 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 222 E 70TH STREET, NEW YORK, NY 10021 SN 0022-1007 J9 J EXP MED JI J. Exp. Med. PD OCT 1 PY 1996 VL 184 IS 4 BP 1279 EP 1284 DI 10.1084/jem.184.4.1279 PG 6 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA VP230 UT WOS:A1996VP23000010 PM 8879199 ER PT J AU Bendelac, A Hunziker, RD Lantz, O AF Bendelac, A Hunziker, RD Lantz, O TI Increased interleukin 4 and immunoglobulin E production in transgenic mice overexpressing NK1T cells SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Article ID NK1.1(+) T-CELLS; CLASS-I MOLECULES; RECEPTOR-ALPHA/BETA(+) CELLS; POSITIVE SELECTION; THYMIC SELECTION; ALPHA-CHAIN; GENE FAMILY; RECEPTOR; THYMOCYTES; ANTIGEN AB Natural Killer (NK)1.1(+) (NK1) T cells are a specialized subset of alpha/beta T cells that coexpress surface receptors that are normally associated with the NK cell lineage of the innate immune system. On recognition of the conserved, major histocompatibility complex class I-like CD1 molecule, these cells are able to release explosive bursts or interleukin 4 (IL-l), a cytokine that promotes the T helper type 2 (Th2) effector class of an immune response, A unique feature of their T cell receptor (TCR) repertoire is the expression of an invariant TCR alpha chain, V alpha 14-J alpha 281, and of a restricted but polyclonal set of V beta gene families, V beta 8, V beta 7, and V beta 2. Here, we show that transgenic expression of this TCR a chain during thymic development is sufficient information to bias the differentiation of mainstream thymocytes towards the NK1 developmental pathway. It markedly increases the frequency of cells with the NK1 pattern of T cell differentiation and also has drastic consequences for the selection of the V beta repertoire. Transgenic CD4 cells exhibited a 10-100-fold increase in IL-4 production on mitogen stimulation in vitro and in vivo, and baseline levels of the Th2-controlled serum immunoglobulin isotypes, IgE and IgG1, were also selectively elevated in vivo. C1 NIAID,TRANSGEN MOUSE FACIL,FREDERICK CANC RES & DEV CTR,CELLULAR & MOL IMMUNOL LAB,NIH,FREDERICK,MD 21702. HOP PAUL BROUSSE,INSERM,U267,F-94807 VILLEJUIF,FRANCE. RP Bendelac, A (reprint author), PRINCETON UNIV,DEPT MOL BIOL,WASHINGTON RD,PRINCETON,NJ 08544, USA. RI Lantz, Olivier/J-4960-2012 OI Lantz, Olivier/0000-0003-3161-7719 NR 40 TC 201 Z9 202 U1 0 U2 0 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 222 E 70TH STREET, NEW YORK, NY 10021 SN 0022-1007 J9 J EXP MED JI J. Exp. Med. PD OCT 1 PY 1996 VL 184 IS 4 BP 1285 EP 1293 DI 10.1084/jem.184.4.1285 PG 9 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA VP230 UT WOS:A1996VP23000011 PM 8879200 ER PT J AU Brown, DR Fowell, DJ Corry, DB Wynn, TA Moskowitz, NH Cheever, AW Locksley, RM Reiner, SL AF Brown, DR Fowell, DJ Corry, DB Wynn, TA Moskowitz, NH Cheever, AW Locksley, RM Reiner, SL TI beta 2-microglobulin-dependent NK1.1(+) T cells are not essential for T helper cell 2 immune responses SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Article ID PROMPTLY PRODUCE INTERLEUKIN-4; SCHISTOSOMA-MANSONI; INTERFERON-GAMMA; LEISHMANIA-MAJOR; IN-VIVO; DEFICIENT MICE; NIPPOSTRONGYLUS-BRASILIENSIS; PHENOTYPE DEVELOPMENT; MURINE LEISHMANIASIS; TRANSGENIC MICE AB A number of investigations have established the critical role of interleukin 4 (IL-4) in mediating the development of T helper (Th)2 effector cells in vitro and in vivo. Despite intensive study, the origin of the IL-4 required for Th2. priming and differentiation remains unclear. Natural killer (NK)1.1(+) alpha/beta T cell receptor(+) T (NT) cells, a unique lineage of cells capable of producing large amounts of IL-4 after activation in vivo, are important candidates for directing Th2 priming. These cells are selected by the nonpolymorphic major histocompatibility complex (MHC) class I molecule, CD1, and are deficient in beta 2-microglobulin (beta 2m)-null mice. We used beta 2m-deficient mice on both BALB/c and C57BL/6 backgrounds to examine their capacity to mount Th2 immune responses after challenge with a number of well-characterized antigens administered by a variety of routes. As assessed by immunization with protein antigen, infection with Leishmania major, embolization with eggs of Schistosoma mansoni, intestinal infection with Nippostrongylus brasiliensis, or induction of airway hyperreactivity to aerosolized antigen, beta 2m-deficient mice developed functional type 2 immune responses that were not substantially different than those in wild-type mice. Production of IL-4 and the generation of immunoglobulin E (IgE) and eosinophil responses were preserved as assessed by a variety of assays. Collectively, these results present a comprehensive analysis of type 2 immune responses in beta 2m-deficient mice, and indicate that beta 2m-dependent NT cells are not required for Th2 development in vivo. C1 UNIV CHICAGO,GWEN KNAPP CTR LUPUS & IMMUNOL RES,COMM IMMUNOL,CHICAGO,IL 60637. UNIV CHICAGO,DEPT MED,CHICAGO,IL 60637. UNIV CALIF SAN FRANCISCO,DEPT MED,SAN FRANCISCO,CA 94143. UNIV CALIF SAN FRANCISCO,DEPT MICROBIOL IMMUNOL,SAN FRANCISCO,CA 94143. NIAID,IMMUNOL & CELL BIOL SECT,PARASIT DIS LAB,NIH,BETHESDA,MD 20892. NIAID,HOST PARASITE RELAT SECT,PARASIT DIS LAB,NIH,BETHESDA,MD 20892. RI Wynn, Thomas/C-2797-2011 FU NIAID NIH HHS [AI-01309, AI-07090, AI-26918] NR 61 TC 174 Z9 175 U1 0 U2 0 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 222 E 70TH STREET, NEW YORK, NY 10021 SN 0022-1007 J9 J EXP MED JI J. Exp. Med. PD OCT 1 PY 1996 VL 184 IS 4 BP 1295 EP 1304 DI 10.1084/jem.184.4.1295 PG 10 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA VP230 UT WOS:A1996VP23000012 PM 8879201 ER PT J AU Vodovotz, Y Lucia, MS Flanders, KC Chesler, L Xie, QW Smith, TW Weidner, J Mumford, R Webber, R Nathan, C Roberts, AB Lippa, CF Sporn, MB AF Vodovotz, Y Lucia, MS Flanders, KC Chesler, L Xie, QW Smith, TW Weidner, J Mumford, R Webber, R Nathan, C Roberts, AB Lippa, CF Sporn, MB TI Inducible nitric oxide synthase in tangle-bearing neurons of patients with Alzheimer's disease SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Article ID GROWTH-FACTOR-BETA; MOLECULAR-CLONING; EXPRESSION; CYTOTOXICITY; TGF-BETA-1; INDUCTION; PROTEIN; BRAINS; CELLS; GAMMA AB In Alzheimer's disease (AD), affected neurons accumulate beta amyloid protein, components of which can induce mouse microglia to express the high-output isoform of nitric oxide synthase (NOS2) in vitro. Products of NOS2 can be neurotoxic. In mice, NOS2 is normally suppressed by transforming growth factor beta 1 (TGF-beta 1). Expression of TGF-beta 1 is decreased in brains from AD patients, a situation that might be permissive for accumulation of NOS2. Accordingly, we investigated the expression of NOS2 in patients with AD, using three monospecific antibodies: a previously described polyclonal and two new monoclonal antibodies. Neurofibrillary tangle-bearing neurons and neuropil threads contained NOS2 in brains from each of 11 AD patients ranging in age from 47 to 81 years, NOS2 was undetectable in brains from 6 control subjects aged 23-72 years, but was expressed in small amounts in 3 control subjects aged 77-87 years. Thus, human neurons can express NOS2 in vivo. The high-output pathway of NO production may contribute to pathogenesis in AD. C1 CORNELL UNIV MED COLL, BEATRICE & SAMUEL A SEAVER LAB, DEPT MED, NEW YORK, NY 10021 USA. UNIV MASSACHUSETTS, MED CTR, DEPT NEUROL, WORCESTER, MA 01655 USA. UNIV MASSACHUSETTS, MED CTR, DEPT PATHOL, WORCESTER, MA 01655 USA. MERCK RES LABS, RAHWAY, NJ 07065 USA. RES & DIAGNOST ANTIBODIES, RICHMOND, CA 94806 USA. MED COLL PENN & HAHNEMANN UNIV, DEPT NEUROL, PHILADELPHIA, PA 19129 USA. RP Vodovotz, Y (reprint author), NCI, RADIAT ONCOL BRANCH,NIH,CHEMOPREVENT LAB,BLDG 10, ROOM B3B69, 9000 ROCKVILLE PIKE, BETHESDA, MD 20892 USA. NR 32 TC 228 Z9 236 U1 0 U2 6 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0022-1007 J9 J EXP MED JI J. Exp. Med. PD OCT 1 PY 1996 VL 184 IS 4 BP 1425 EP 1433 DI 10.1084/jem.184.4.1425 PG 9 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA VP230 UT WOS:A1996VP23000025 PM 8879214 ER PT J AU Chitnis, CE Chaudhuri, A Horuk, R Pogo, AO Miller, LH AF Chitnis, CE Chaudhuri, A Horuk, R Pogo, AO Miller, LH TI The domain on the Duffy blood group antigen for binding Plasmodium vivax and P-knowlesi malarial parasites to erythrocytes SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Article ID GROUP SYSTEM; RECEPTOR; FALCIPARUM; GLYCOPROTEIN; EXPRESSION; PROTEINS; INVASION; GENE; IDENTIFICATION; MEROZOITES AB Plasmodium vivax and the related simian malarial parasite P. krowlesi use the Duffy blood group antigen as a receptor to invade human erythrocytes and region II of the parasite ligands for binding to this erythrocyte receptor. Here, we identify the peptide within the Duffy blood group antigen of human and rhesus erythrocytes to which the P. vivax and P. knowlesi ligands bind. Peptides from the NH2-terminal extracellular region of the Duffy antigen were tested for their ability to block the binding of erythrocytes to transfected Cos cells expressing on their surface region II of the Duffy-binding ligands. The binding site on the human Duffy antigen used by both the P. vivax and P. knowlesi ligands maps to a 35-amino acid region. A 34-amino acid peptide from the equivalent region of the rhesus Duffy antigen blocked the binding of P. vivax to human erythrocytes, although the P. vivax ligand expressed on Cos cells does not bind rhesus erythrocytes. The binding of the rhesus peptide, but not the rhesus erythrocyte, to the P. vivax ligand was explained by interference of carbohydrate with the binding process. Rhesus erythrocytes, treated with N-glycanase, bound specifically to P. vivax region II. Thus, the interaction of P. vivax ligand with human and rhesus erythrocytes appears to be mediated by a peptide-peptide interaction. Glycosylation of the rhesus Duffy antigen appears to block binding of the P. vivax ligand to rhesus erythrocytes. C1 NIAID,PARASIT DIS LAB,NIH,BETHESDA,MD 20892. NEW YORK BLOOD CTR,DEPT CELL BIOL,NEW YORK,NY 10021. BERLEX BIOSCI INC,DEPT IMMUNOL,RICHMOND,CA 94804. FU NHLBI NIH HHS [HL 53927] NR 20 TC 112 Z9 115 U1 0 U2 4 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 222 E 70TH STREET, NEW YORK, NY 10021 SN 0022-1007 J9 J EXP MED JI J. Exp. Med. PD OCT 1 PY 1996 VL 184 IS 4 BP 1531 EP 1536 DI 10.1084/jem.184.4.1531 PG 6 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA VP230 UT WOS:A1996VP23000036 PM 8879225 ER PT J AU Peruzzi, M Wagtmann, N Long, EO AF Peruzzi, M Wagtmann, N Long, EO TI A p70 killer cell inhibitory receptor specific for several HLA-B allotypes discriminates among peptides bound to HLA-B*2705 SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Article ID MHC CLASS-I; NK CLONES; RECOGNITION; MOLECULES; LYSIS; EXPRESSION; PROTECTION; ALLELES; CLONING; EPITOPE AB Natural killer (NK) cells express a repertoire of killer cell inhibitory receptors (KIR) for major histocompatibility complex (MHC) class I molecules. KIR specificity for MHC class I can be broad, as in the case of a single p70 KIR that can recognize several HLA-B allotypes, including HLA-B*2705. On the other hand, recognition of MHC class I can also be highly specific, as in the case of NK clones that recognize HLA-B*2705 in a peptide-specific manner. Most NK cells express multiple KIR sequences. To determine whether the broad and specific types of HLA-B recognition by NK cells reflect the use of different receptors or a property of a single KIR we analyzed the recognition of HLA-B*2705 by the p70 KIR-11, known to recognize several HLA-B allotypes. Vaccinia virus-mediated expression of KIR-11 in NK clones resulted in inhibition by HLA-B*2705 molecules on wild type but not on target cells deficient in the transporter for antigen presentation (TAP). Two peptides (FRYNGLIHR and RRSKEITVR) loaded onto HLA-B*2705 molecules on TAP-deficient cells provided protection from lysis by NK cells expressing KIR-11 but three other B27-specific peptides did not. As the five peptides bound to HLA-B*2705 with similar stability, these data demonstrate that a single KIR specific for several HLA-B allotypes recognizes a subset of peptides bound to HLA-B*2705. C1 NIAID,IMMUNOGENET LAB,NIH,ROCKVILLE,MD 20852. RI Long, Eric/G-5475-2011 OI Long, Eric/0000-0002-7793-3728 NR 26 TC 102 Z9 104 U1 0 U2 5 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 222 E 70TH STREET, NEW YORK, NY 10021 SN 0022-1007 J9 J EXP MED JI J. Exp. Med. PD OCT 1 PY 1996 VL 184 IS 4 BP 1585 EP 1590 DI 10.1084/jem.184.4.1585 PG 6 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA VP230 UT WOS:A1996VP23000045 PM 8879234 ER PT J AU Fujita, K Maldarelli, F Silver, J AF Fujita, K Maldarelli, F Silver, J TI Bimodal down-regulation of CD4 in cells expressing human immunodeficiency virus type 1 Vpu and Env SO JOURNAL OF GENERAL VIROLOGY LA English DT Article ID MURINE LEUKEMIA-VIRUS; FV-4 RESISTANCE GENE; ENVELOPE PROTEIN; CYTOPLASMIC DOMAIN; INTERFERENCE; DEGRADATION; MICE; INFECTIVITY; RETROVIRUS; MEMBRANE AB We analysed clones of HeLa cells stably expressing the human immunodeficiency virus (HIV-1) envelope gene (env) and the HIV-1 receptor, CD4, Surprisingly, individual clones were found to consist of two distinct populations of cells differing by about 10-fold in the level of surface CD4, When high and low CD4-expressing cells were separated by FACS, each subpopulation gave rise to a mixture of high and low CD4-expressing cells after several days in culture, High and low CD4-expressing subpopulations did not differ with respect to the amount of intracellular Env, but there was an inverse correlation between CD4 and another HIV-1 protein encoded by the same segment of the HIV genome, Vpu, High surface CD4 cells had high levels of intracellular CD4, largely in the perinuclear region, and low levels of Vpu with a diffuse staining pattern, Conversely, low surface CD4 cells had low levels of intracellular CD4 with a diffuse staining pattern, and high levels of Vpu, largely in the perinuclear region, Vectors containing mutant versions of either Env or Vpu failed to down-regulate surface CD4, The phenomenon of bimodal expression of a surface protein in cells derived from single clones provides a simple model of differentiation in vitro, We show how a hypothetical interaction between CD4 and a multimer of Vpu, the multimerization of which is cooperative, would lead to bimodal expression of CD4, This model may be generalized and could explain other cellular 'switches'. C1 NIAID,MOL MICROBIOL LAB,NIH,BETHESDA,MD 20892. NR 29 TC 6 Z9 6 U1 0 U2 0 PU SOC GENERAL MICROBIOLOGY PI READING PA HARVEST HOUSE 62 LONDON ROAD, READING, BERKS, ENGLAND RG1 5AS SN 0022-1317 J9 J GEN VIROL JI J. Gen. Virol. PD OCT PY 1996 VL 77 BP 2393 EP 2401 DI 10.1099/0022-1317-77-10-2393 PN 10 PG 9 WC Biotechnology & Applied Microbiology; Virology SC Biotechnology & Applied Microbiology; Virology GA VL319 UT WOS:A1996VL31900002 PM 8887470 ER PT J AU Russell, P Johnson, DH AF Russell, P Johnson, DH TI Enzymes protective of oxidative damage present in all decades of life in the trabecular meshwork, as detected by two-dimensional gel electrophoresis protein maps SO JOURNAL OF GLAUCOMA LA English DT Article DE oxidative damage; enzymes; protection; age-related differences; two-dimensional gel electrophoresis; trabecular meshwork; proteins; cornea; sclera ID AGE-RELATED-CHANGES; AQUEOUS-HUMOR; HYDROGEN-PEROXIDE; OUTFLOW FACILITY; CALF; GLUTATHIONE; LOCALIZATION; GLAUCOMA; EYES AB Our purpose was to determine whether there are age-related differences in the presence of enzymes protective against oxidative damage in normal human trabecular meshwork (TM). The study was also done to determine whether there are significant changes in proteins related to age, gender, or position around the circumference of the meshwork. Human TM was dissected from both eyes of 16 normal donors ranging in age from 9 to 91 years. Proteins were mapped with two-dimensional (2D) gel electrophoresis. The microanalytical method used allowed analysis of samples as small as one fourth of a single meshwork. Selected proteins were identified by Western blot. Superoxide dismutase (SOD) and glutathione reductase were present in all ages on the 2D maps. Age-related changes in the TM were not apparent, although slight differences may exist between juvenile and adult TM, since differences in the relative intensities of some polypeptides were evident. In any single quadrant of the TM, greater than or equal to 160 protein spots could be counted visually. The 2D maps of the various quadrants were very similar, and the protein patterns were analogous between fellow eyes. Distinct differences were noted between maps from TM and adjacent cornea and sclera. Other protein spots identified included actin, albumin, transferrin, and vimentin. The TM samples had a mean cellular protein content of 66.3 +/- 15.4 mu g. C1 MAYO CLIN,ROCHESTER,MN. RP Russell, P (reprint author), NATL INST HLTH,MSC 2735,BLDG 6,RM 228,BETHESDA,MD 20892, USA. NR 29 TC 28 Z9 29 U1 0 U2 1 PU LIPPINCOTT-RAVEN PUBL PI PHILADELPHIA PA 227 EAST WASHINGTON SQ, PHILADELPHIA, PA 19106 SN 1057-0829 J9 J GLAUCOMA JI J. Glaucoma PD OCT PY 1996 VL 5 IS 5 BP 317 EP 324 PG 8 WC Ophthalmology SC Ophthalmology GA VL342 UT WOS:A1996VL34200006 PM 8897231 ER PT J AU Seder, RA Kelsall, BL Jankovic, D AF Seder, RA Kelsall, BL Jankovic, D TI Differential roles for IL-12 in the maintenance of immune responses in infectious versus autoimmune disease SO JOURNAL OF IMMUNOLOGY LA English DT Article ID IFN-GAMMA; LEISHMANIA-MAJOR; INTERFERON-GAMMA; CELL RESPONSE; T-CELLS; B-CELLS; INTERLEUKIN-12; MICE; ENCEPHALOMYELITIS; PROLIFERATION AB IL-12 has been shown to be important in the generation of a functional Th1 response in animal models of both infectious and autoimmune disease, Furthermore, the role of IL-12 in the maintenance of the immune responses in these diseases is now emerging, Herein, we discuss the idea that memory responses for certain infections may be IL-12 independent, whereas memory responses for specific autoimmune diseases still require IL-12 to maintain a pathogenic response. C1 NIAID,PARASIT DIS LAB,NIH,BETHESDA,MD 20892. RP Seder, RA (reprint author), NIAID,CLIN INVEST LAB,NIH,BLDG 10,ROOM 11C215,9000 ROCKVILLE PIKE,BETHESDA,MD 20892, USA. NR 24 TC 98 Z9 99 U1 0 U2 0 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD OCT 1 PY 1996 VL 157 IS 7 BP 2745 EP 2748 PG 4 WC Immunology SC Immunology GA VL106 UT WOS:A1996VL10600001 PM 8816374 ER PT J AU Bader, T Nettesheim, P AF Bader, T Nettesheim, P TI Tumor necrosis factor-alpha modulates the expression of its p60 receptor and several cytokines in rat tracheal epithelial cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID EPIDERMAL GROWTH-FACTOR; NITRIC-OXIDE SYNTHASE; INTERFERON-GAMMA; DOWN-REGULATION; HUMAN-FIBROBLASTS; MESSENGER-RNA; TNF RECEPTORS; LUNG-TISSUE; MURINE; INDUCTION AB The epithelium of the conducting airways is frequently the target of toxic chemical and microbial agents causing inflammation, hypersecretion, and epithelial necrosis. TNF-alpha is a prototypical inflammatory cytokine released by macrophages and other inflammatory cells. The purpose of this study was to characterize TNF-alpha receptors in fully differentiated rat tracheal epithelial (RTE) cells in culture and to examine the effects of TNF-alpha on this epithelium. We demonstrated the presence of approximately 250 TNF-alpha receptors per RTE cell. Both known receptor types, p60 (TNF-RI, CD 120a) and p80 (TNF-RII, CD 120b), were expressed. The level of p80 mRNA was unaffected by TNF-alpha treatment, whereas p60 mRNA was down-regulated, and soluble TNF-RI was shed from cells within 30 min. Treatment of RTE cultures with TNF-alpha (1000 U) caused no cytotoxicity (as determined by lactate dehydrogenase release). However, TNF-alpha exposure of the cultures induced the expression of several inflammatory mediators, as determined by reverse transcription-PCR and ELISA. Low levels of IFN-gamma mRNA became detectable after 4 h. Increased levels of TNF-alpha mRNA and protein were found, which peaked after 6 h of TNF-alpha treatment, but neither IL-1 alpha nor IL-1 beta was detectable. Calcium-independent nitric oxide;synthase transcripts were elevated two- to threefold within 2 to 6 h of TNF-alpha treatment. These findings suggest that the airway epithelium may actively participate in the pathogenesis of airway inflammation through the production of mediators similar to those found in a Th1 response. RP Bader, T (reprint author), NIEHS,111 TW ALEXANDER DR,RES TRIANGLE PK,NC 27709, USA. NR 57 TC 25 Z9 25 U1 0 U2 1 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD OCT 1 PY 1996 VL 157 IS 7 BP 3089 EP 3096 PG 8 WC Immunology SC Immunology GA VL106 UT WOS:A1996VL10600046 PM 8816419 ER PT J AU Thomas, DL Shih, JW Alter, HJ Vlahov, D Cohn, S Hoover, DR Cheung, L Nelson, KE AF Thomas, DL Shih, JW Alter, HJ Vlahov, D Cohn, S Hoover, DR Cheung, L Nelson, KE TI Effect of human immunodeficiency virus on hepatitis C virus infection among injecting drug users SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID NON-B-HEPATITIS; SEXUALLY-TRANSMITTED DISEASES; POLYMERASE CHAIN-REACTION; RNA LEVELS; NON-A; HEMODIALYSIS-PATIENTS; HEMOPHILIA PATIENTS; NATURAL-HISTORY; LIVER-DISEASE; HCV INFECTION AB To assess the effect of human inmunodeficiency virus (HIV) immunosuppression on ongoing hepatitis C virus (HCV) infection, CD4 lymphocyte counts and serum concentrations of HCV RNA, HIV RNA, and alanine aminotransferase (ALT) were evaluated among members of a cohort of injecting drug users (IDUs). With 100 participants randomly selected at various stages of HIV-related immunosuppression, serum HCV RNA concentrations increased with age (P = .007) and were higher in HIV-positive IDUs with 201-500 (P = .026) and 51-200 (P = .004) CD4 cells/mL than in HN-negative participants. Among 27 HCV-infected IDUs who acquired HIV infection, serum HCV RNA concentrations varied between semiannual visits by a mean of 0.45 logs, increasing by 0.60 logs after HIV seroconversion (P < .0001), by 0.12 logs each subsequent year (P = .006), and by 0.36 logs per log increase in CD4 cells (P = .01). Serum ALT levels were similar between HIV-positive (40.1 IU/mL) and HIV-negative (45.4 IU/mL) patients (P > .10). While HIV infection and possibly HIV progression are associated with increased HCV RNA levels, other factors appear to affect biochemical and virologic markers of HCV infection in some dually infected persons. C1 JOHNS HOPKINS UNIV,SCH MED,DIV INFECT DIS,BALTIMORE,MD 21205. JOHNS HOPKINS UNIV,SCH HYG & PUBL HLTH,DEPT EPIDEMIOL,BALTIMORE,MD. NIH,DEPT TRANSFUS MED,BALTIMORE,MD. FU NIDA NIH HHS [DA-23201, DA-04334, DA-05911] NR 43 TC 178 Z9 180 U1 2 U2 2 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD OCT PY 1996 VL 174 IS 4 BP 690 EP 695 PG 6 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA VK045 UT WOS:A1996VK04500002 PM 8843204 ER PT J AU Welles, SL Jackson, JB YenLieberman, B Demeter, L Japour, AJ Smeaton, LM Johnson, VA Kuritzkes, DR DAquila, RT Reichelderfer, PA Richman, DD Reichman, R Fischl, M Dolin, R Coombs, RW Kahn, JO McLaren, C Todd, J Kwok, S AF Welles, SL Jackson, JB YenLieberman, B Demeter, L Japour, AJ Smeaton, LM Johnson, VA Kuritzkes, DR DAquila, RT Reichelderfer, PA Richman, DD Reichman, R Fischl, M Dolin, R Coombs, RW Kahn, JO McLaren, C Todd, J Kwok, S TI Prognostic value of plasma human immunodeficiency virus type 1 (HIV-1) RNA levels in patients with advanced HIV-1 disease and with little or no prior zidovudine therapy SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID SYNCYTIUM-INDUCING PHENOTYPE; REVERSE-TRANSCRIPTASE; CLINICAL-TRIALS; PROGRESSION; RESISTANCE; INFECTION; QUANTITATION; DIDANOSINE; SENSITIVITY; MUTATION AB The association of plasma human immunodeficiency virus type 1 (HIV-1) RNA level at study entry and over time with clinical progression was evaluated in 187 patients from AIDS Clinical Trials Group protocol 116A who had little or no prior zidovudine treatment. Three-fold-higher HIV-1 RNA levels at study entry and 3-fold increases by week 8 were associated with progression (relative hazard [RH], 1.67; 95% confidence limits [CL], 1.20, 2.32; and RH, 1.45; CL, 1.02, 2.05, respectively). Having 3-fold-higher CD4 cell count at entry was independently associated with a 52% reduction in risk for progression (adjusted RH, 0.48; CL, 0.33, 0.70). then stratified by length of prior zidovudine therapy, RNA level was predictive in drug-naive patients (adjusted RH, 1.87; CL, 1.23, 2.85) but not predictive in patients with up to 16 weeks of prior therapy (adjusted RH, 1.11; CL, 0.70, 1.76). Analysis suggests that the acquisition of mutations at HIV-1 reverse transcriptase codons 215 and 74 is associated with subsequent increases in HIV-1 RNA level (relative risk, 7.00; CL, 0.86, 56.90). C1 HARVARD UNIV,SCH PUBL HLTH,CTR BIOSTAT AIDS RES,BOSTON,MA 02115. HARVARD UNIV,SCH PUBL HLTH,DEPT BIOSTAT,BOSTON,MA 02115. HARVARD UNIV,BETH ISRAEL HOSP,SCH MED,BOSTON,MA. HARVARD UNIV,MASSACHUSETTS GEN HOSP,SCH MED,BOSTON,MA. CASE WESTERN RESERVE UNIV,INST PATHOL,CLEVELAND,OH 44106. UNIV HOSP CLEVELAND,CLEVELAND,OH. CLEVELAND CLIN FDN,CLEVELAND,OH 44195. UNIV ROCHESTER,DEPT MED,ROCHESTER,NY. UNIV ALABAMA,DEPT MED,DIV INFECT DIS,BIRMINGHAM,AL 35294. VET AFFAIRS MED CTR,BIRMINGHAM,AL. UNIV COLORADO,HLTH SCI CTR,DENVER,CO. VET AFFAIRS MED CTR,DENVER,CO. NIAID,NATL INST HLTH,DIV AIDS,BETHESDA,MD 20892. UNIV CALIF SAN DIEGO,SAN DIEGO,CA 92103. SAN DIEGO VET AFFAIRS MED CTR,LA JOLLA,CA. UNIV CALIF SAN FRANCISCO,SAN FRANCISCO,CA 94143. SAN FRANCISCO GEN HOSP,AIDS PROGRAM,SAN FRANCISCO,CA 94110. CHIRON CORP,EMERYVILLE,CA 94608. ROCHE MOL SYST,ALAMEDA,CA. UNIV MIAMI,DEPT MED,MIAMI,FL. UNIV WASHINGTON,DEPT LAB MED,SEATTLE,WA 98195. BRISTOL MYERS SQUIBB,WALLINGFORD,CT. FU NIAID NIH HHS [AI-25879, AI-01101, AI-05030] NR 34 TC 82 Z9 83 U1 0 U2 2 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD OCT PY 1996 VL 174 IS 4 BP 696 EP 703 PG 8 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA VK045 UT WOS:A1996VK04500003 PM 8843205 ER PT J AU Coombs, RW Welles, SL Hooper, C Reichelderfer, PS DAquila, RT Japour, AJ Johnson, VA Kuritzkes, DR Richman, DD Kwok, S Todd, J Jackson, JB DeGruttola, V Crumpacker, CS Kahn, J AF Coombs, RW Welles, SL Hooper, C Reichelderfer, PS DAquila, RT Japour, AJ Johnson, VA Kuritzkes, DR Richman, DD Kwok, S Todd, J Jackson, JB DeGruttola, V Crumpacker, CS Kahn, J TI Association of plasma human immunodeficiency virus type 1 RNA level with risk of clinical progression in patients with advanced infection SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID POLYMERASE CHAIN-REACTION; DISEASE PROGRESSION; ANTIRETROVIRAL THERAPY; ZIDOVUDINE RESISTANCE; COMBINATION THERAPY; HIV-1 INFECTION; DIDANOSINE; VIREMIA; AIDS; QUANTITATION AB Human immunodeficiency virus (HIV)-1 RNA level in plasma was evaluated as a surrogate marker for disease progression in a clinical trial of advanced HIV-1 infection, Baseline MV-I RNA level was an independent predictor of disease progression (relative hazard [RH] for each doubling of HIV-1 RNA level, 1.26; 95% confidence interval [CI], 1.03-1.54; P = .02), after adjusting for the week 4 change in HIV-1 RNA level, baseline CD4 cell count, syncytium-inducing phenotype, clinical status at study entry, and therapy randomization. A 50% reduction in HIV-1 RNA level was associated with a 27% decrease in the adjusted risk of disease progression during the study (RH, 0.73; 95% CI, 0.52-1.02; P = .07). The partial validation of HIV-1 RNA as a predictor for clinical end points has implications for the use of HIV-1 RNA in clinical trials and practice. C1 HARVARD UNIV,SCH PUBL HLTH,CTR BIOSTAT AIDS RES,BOSTON,MA 02115. HARVARD UNIV,MASSACHUSETTS GEN HOSP,SCH MED,BOSTON,MA. HARVARD UNIV,BETH ISRAEL HOSP,SCH MED,BOSTON,MA. NIAID,NATL INST HLTH,DIV AIDS,BETHESDA,MD 20892. UNIV ALABAMA,SCH MED,BIRMINGHAM,AL. VET AFFAIRS MED CTR,BIRMINGHAM,AL. UNIV COLORADO,HLTH SCI CTR,DENVER,CO. VET AFFAIRS MED CTR,DENVER,CO. UNIV CALIF SAN DIEGO,SAN DIEGO,CA 92103. VET AFFAIRS MED CTR,SAN DIEGO,CA 92161. UNIV CALIF SAN FRANCISCO,SAN FRANCISCO,CA 94143. ROCHE MOL SYST,ALAMEDA,CA. CHIRON CORP,EMERYVILLE,CA 94608. CASE WESTERN RESERVE UNIV,CLEVELAND,OH 44106. UNIV HOSP CLEVELAND,CLEVELAND,OH 44106. RP Coombs, RW (reprint author), UNIV WASHINGTON,VIROL OFF,ROOM 9301,PACIFIC MED CTR,1200 12TH AVE,SEATTLE,WA 98144, USA. FU NIAID NIH HHS [AI-27664, AI-27757, AI-05030] NR 50 TC 151 Z9 153 U1 0 U2 4 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD OCT PY 1996 VL 174 IS 4 BP 704 EP 712 PG 9 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA VK045 UT WOS:A1996VK04500004 PM 8843206 ER PT J AU Bertolli, J StLouis, ME Simonds, RJ Nieburg, P Kamenga, M Brown, C Tarande, M Quinn, T Ou, CY AF Bertolli, J StLouis, ME Simonds, RJ Nieburg, P Kamenga, M Brown, C Tarande, M Quinn, T Ou, CY TI Estimating the timing of mother-to-child transmission of human immunodeficiency virus in a breast-feeding population in Kinshasa, Zaire SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID POLYMERASE CHAIN-REACTION; HIV-1; TYPE-1 AB Breast-fed infants born to human immunodeficiency virus (HIV)-infected mothers in Kinshasa, Zaire, were monitored a mean of 18 months. HIV infection in infants was determined by polymerase chain reaction (PCR), HIV culture, or ELISA. PCR test results for HIV DNA on venous blood drawn from children ages 0-2 days and 3-5 months were used to estimate proportions of mother-to-child transmission and transmission risks during the intrauterine, intrapartum/early postpartum, and late postpartum periods. Among 69 HIV-infected children (26% of the cohort), 23% (95% confidence interval [CI], 14%-35%) were estimated to have had intrauterine, 65% (CI, 53%-76%) intrapartum/early postpartum, and 12% (CI, 5%-22%) late postpartum transmission. The estimated risks for intrauterine, intrapartum/early postpartum, and late postpartum infection, respectively, were 6% (16/261; CI, 4%-10%), 18% (45/245; CI, 14%-24%), acid 4% (8/189; CI, 2%-8%). These results support earlier studies indicating that most transmission occurs during labor and delivery or in the early postpartum period and that the risk of HIV transmission through breast-feeding during the postpartum period is substantial. C1 CTR DIS CONTROL & PREVENT,NATL CTR HIV SEXUALLY TRANSMIT DIS & TB PREVENT,ATLANTA,GA 30341. CTR DIS CONTROL & PREVENT,NATL CTR INFECT DIS,DIV AIDS SEXUALLY TRANSMITTED DIS & TB LAB RES,ATLANTA,GA. PROJET SIDA,KINSHASA,ZAIRE. NIAID,NATL INST HLTH,BETHESDA,MD 20892. RP Bertolli, J (reprint author), CTR DIS CONTROL & PREVENT,DIV HIV AIDS PREVENT,1600 CLIFTON RD,MS E-45,ATLANTA,GA 30333, USA. NR 9 TC 126 Z9 133 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD OCT PY 1996 VL 174 IS 4 BP 722 EP 726 PG 5 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA VK045 UT WOS:A1996VK04500006 PM 8843208 ER PT J AU Weiss, SM Roblin, PM Gaydos, CA Quinn, TC Hammerschlag, MR AF Weiss, SM Roblin, PM Gaydos, CA Quinn, TC Hammerschlag, MR TI Detection of Chlamydia pneumoniae in atheroma specimens - Reply SO JOURNAL OF INFECTIOUS DISEASES LA English DT Letter ID CORONARY-ARTERY DISEASE; HEART-DISEASE; TWAR; ASSOCIATION; INFECTION C1 SUNY HLTH SCI CTR,DEPT PEDIAT,BROOKLYN,NY 11203. JOHNS HOPKINS UNIV,SCH MED,DEPT MED,DIV INFECT DIS,BALTIMORE,MD 21205. NIAID,IMMUNOREGULAT LAB,BETHESDA,MD 20892. RP Weiss, SM (reprint author), SUNY HLTH SCI CTR,DEPT MED,DIV INFECT DIS,450 CLARKSON AVE,BOX 56,BROOKLYN,NY 11203, USA. RI Gaydos, Charlotte/E-9937-2010 NR 11 TC 2 Z9 2 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 S WOODLAWN AVE, CHICAGO, IL 60637 SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD OCT PY 1996 VL 174 IS 4 BP 895 EP 896 PG 2 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA VK045 UT WOS:A1996VK04500040 ER PT J AU Luo, H Lu, YD Mao, Y Shi, XL Dalal, NS AF Luo, H Lu, YD Mao, Y Shi, XL Dalal, NS TI Role of chromium(IV) in the chromium(VI)-related free radical formation, dG hydroxylation, and DNA damage SO JOURNAL OF INORGANIC BIOCHEMISTRY LA English DT Article ID ONE-ELECTRON REDUCTION; HYDROGEN-PEROXIDE; GLUTATHIONE-REDUCTASE; SODIUM CHROMATE(VI); CARCINOGEN CHROMATE; PARAMAGNETIC CHROMIUM; METAL CARCINOGENESIS; PRINCIPAL REDUCTANT; CR(V) COMPLEXES; ASCORBIC-ACID AB A reactive Cr(IV) ester was synthesized from a reaction of CrO3 with 2,4-dimethyl-2,4-pentanediol as verified by electron spin resonance (ESR) and magnetic susceptibility measurements. ESR spin trapping studies demonstrate that the Cr(IV) ester is capable of generating hydroxyl free radical ((OH)-O-.) through a Fenton-like mechanism (Cr(IV) + H2O2 --> Cr(V)+ (OH)-O-. + OH-) with a concomitant generation of Cr(V) species (g = 1.9787). Cr(IV) caused DNA strand breaks as measured by electrophoretic assays. H2O2 enhanced the DNA strand breaks via (OH)-O-. formation by a Cr(IV)-mediated Fenton-like reaction. In the Cr(IV)/H2O2 system, formate did not block Cr(V) formation, but prevented DNA damage, indicating that (OH)-O-. radicals, and not Cr(V), caused the DNA damage. Reaction of Cr(VI) with ascorbate was also used as a source of Cr(IV). Incubation of Cr(VI), ascorbate, and DNA caused DNA strand breaks. A free radical trap, 5,5-dimethyl-1-pyrroline (DMPO), only slightly inhibited the DNA damage. Addition of Mn(II), which inhibited Cr(IV), caused significant protection. H2O2 enhanced the DNA damage via Cr(IV)-mediated (OH)-O-. radical generation and Mn(II) inhibited the damage, again showing that Cr(IV) and its related (OH)-O-. generation caused DNA strand breaks. HPLC measurements showed that (OH)-O-. radicals generated by a Cr(IV)-mediated Fenton-like reaction generated 8-hydroxy-2'-deoxyguanosine from 2'-deoxyguanosine. The results demonstrate that Cr(IV) and its generated (OH)-O-. radicals are capable of damaging DNA. Moreover, in comparison with Cr(V), Cr(IV) is a more potent DNA damaging agent. C1 NCI,LAB EXPT PATHOL,BETHESDA,MD 20892. W VIRGINIA UNIV,DEPT CHEM,MORGANTOWN,WV 26506. RI Shi, Xianglin/B-8588-2012 NR 49 TC 32 Z9 33 U1 1 U2 4 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 SN 0162-0134 J9 J INORG BIOCHEM JI J. Inorg. Biochem. PD OCT PY 1996 VL 64 IS 1 BP 25 EP 35 DI 10.1016/0162-0134(95)00241-3 PG 11 WC Biochemistry & Molecular Biology; Chemistry, Inorganic & Nuclear SC Biochemistry & Molecular Biology; Chemistry GA VJ787 UT WOS:A1996VJ78700003 PM 8837499 ER PT J AU Moriwaki, SI Stefanini, M Lehmann, AR Hoeijmakers, JHJ Robbins, JH Rapin, I Botta, E Tanganelli, B Vermeulen, W Broughton, BC Kraemer, KH AF Moriwaki, SI Stefanini, M Lehmann, AR Hoeijmakers, JHJ Robbins, JH Rapin, I Botta, E Tanganelli, B Vermeulen, W Broughton, BC Kraemer, KH TI DNA repair and ultraviolet mutagenesis in cells from a new patient with xeroderma pigmentosum group G and Cockayne syndrome resemble xeroderma pigmentosum cells SO JOURNAL OF INVESTIGATIVE DERMATOLOGY LA English DT Article DE shuttle vector plasmid; skin cancer; transfection; photoproducts ID COMPLEMENTATION GROUP-G; EXCISION-REPAIR; INCREASED SENSITIVITY; CELLULAR-SENSITIVITY; DIMER PHOTOPRODUCTS; CLINICAL-FEATURES; UV IRRADIATION; RNA-SYNTHESIS; GENE; DEFECT AB Xeroderma pigmentosum (XP)/Cockayne syndrome (CS) complex is a combination of clinical features of two rare genetic disorders in one individual, A sun-sensitive boy (XP20BE) who had severe symptoms of CS, with dwarfism, microcephaly, retinal degeneration, and mental impairment, had XP-type pigmentation and died at 6 y with marked cachexia (weight 14.5 lb) without skin cancers, We evaluated his cultured cells for characteristic CS or XP DNA-repair abnormalities. The level of ultraviolet (UV)-induced unscheduled DNA synthesis was less than 5% of normal, characteristic of the excision-repair defect of XP. Cell fusion studies indicated that his cells were in XP complementation group G, His cells were hypersensitive to killing by UV, and their post-UV recovery of RNA synthesis was abnormally low, features of both CS and XP, Post-UV survival of plasmid pSP189 in his cells was markedly reduced, and post-UV plasmid mutation frequency was higher than with normal cells, as in both CS and XP, Sequence analysis of the mutated plasmid marker gene showed normal frequency of plasmids with multiple base substitutions, as in CS, and an abnormally increased frequency of G:C --> A:T mutations, a feature of XP. Transfection of UV-treated pRSV cat with or without photoreactivation revealed that his cells, like XP cells, could not repair either cyclobutane pyrimidine dimers or non-dimer photoproducts. These results indicate that the DNA-repair features of the XP20BE (XP-G/CS) cells are phenotypically more like XP cells than CS cells, whereas clinically the CS phenotype is more prominent than XP. C1 NCI,MOL CARCINOGENESIS LAB,BETHESDA,MD 20892. NCI,DERMATOL BRANCH,BETHESDA,MD 20892. CNR,IST GENET BIOCHIM EVOLUZ,PAVIA,ITALY. UNIV SUSSEX,MRC,CELL MUTAT UNIT,BRIGHTON BN1 9RR,E SUSSEX,ENGLAND. ERASMUS UNIV ROTTERDAM,MED GENET CTR,DEPT CELL BIOL & GENET,ROTTERDAM,NETHERLANDS. ALBERT EINSTEIN COLL MED,DEPT NEUROL,BRONX,NY 10467. RI Botta, Elena/B-9065-2015 FU Telethon [E.0197] NR 49 TC 42 Z9 43 U1 0 U2 1 PU BLACKWELL SCIENCE INC PI CAMBRIDGE PA 238 MAIN ST, CAMBRIDGE, MA 02142 SN 0022-202X J9 J INVEST DERMATOL JI J. Invest. Dermatol. PD OCT PY 1996 VL 107 IS 4 BP 647 EP 653 DI 10.1111/1523-1747.ep12584287 PG 7 WC Dermatology SC Dermatology GA VJ503 UT WOS:A1996VJ50300023 PM 8823375 ER PT J AU Johnston, JA Bacon, CM Riedy, MC OShea, JJ AF Johnston, JA Bacon, CM Riedy, MC OShea, JJ TI Signaling by IL-2 and related cytokines: JAKs, STATs, and relationship to immunodeficiency SO JOURNAL OF LEUKOCYTE BIOLOGY LA English DT Review DE cytokine signaling; signal transduction; IL-4; IL-12; IL-15; severe combined immunodeficiency; receptor; Janus; JAK3; STAT5 ID PROTEIN-TYROSINE KINASE; RECEPTOR-GAMMA-CHAIN; HUMAN INTERLEUKIN-4 RECEPTOR; GROWTH-HORMONE RECEPTOR; DNA-BINDING ACTIVITY; MICE LACKING JAK3; C-MYC INDUCTION; BETA-CHAIN; JANUS KINASE; TRANSDUCTION PATHWAY AB Cytokines that bind to the interleukin-2 (IL-2) receptor common gamma chain (gamma c), including IL-2, IL-4, IL-7, IL-9, and IL-15, are important for the growth and differentiation of T and B lymphocytes, natural killer cells, macrophages, and monoctyes. These cytokines have overlapping biological effects that in part result from the use of the shared receptor subunit gamma c. Recently it has become clear that these cytokines activate a number of important intracellular signaling molecules, including the Janus kinases JAK1 and JAK3 and members of the transcription factor family of signal transducers and activators of transcription (STATs). The discovery of these signaling pathways has led to important new insights into their role in lymphocyte maturation, as it has emerged that mutations in the genes encoding both gamma c and JAK3 result in similar forms of severe combined immunodeficiency (SCID), In this review we examine the structure and function of cytokine receptors and the signaling pathways involved in their regulation of gene expression. Furthermore, we discuss recent advances that have led to a better understanding of how cytokines elicit intracellular responses, as well as their role in normal lymphoid development. RP Johnston, JA (reprint author), NIAMSD,LYMPHOCYTE CELL BIOL SECT,NIA,BLDG 10,ROOM 9N228,10 CTR DR,MSC 1820,BETHESDA,MD 20892, USA. NR 156 TC 82 Z9 82 U1 0 U2 0 PU FEDERATION AMER SOC EXP BIOL PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 SN 0741-5400 J9 J LEUKOCYTE BIOL JI J. Leukoc. Biol. PD OCT PY 1996 VL 60 IS 4 BP 441 EP 452 PG 12 WC Cell Biology; Hematology; Immunology SC Cell Biology; Hematology; Immunology GA VM935 UT WOS:A1996VM93500002 PM 8864127 ER PT J AU Leonard, EJ Skeel, A AF Leonard, EJ Skeel, A TI Hepatic catabolism of intravenously administered pro-macrophage-stimulating protein in mice SO JOURNAL OF LEUKOCYTE BIOLOGY LA English DT Article DE pharmacokinetics; plasma; clearance; radioactivity; proteolysis ID HEPATOCYTE GROWTH-FACTOR; AMINO-ACID-SEQUENCE; PLASMA-CLEARANCE; TYROSINE KINASE; HUMAN-SERUM; MSP; RECEPTOR; MOUSE; IDENTIFICATION; BINDING AB We injected I-125-pro-macrophage-stimulating protein (pro-MSP) intravenously into normal mice to determine its clearance from the circulation and to test for conversion of pro-MSP to the biologically active heterodimer in the absence of inflammation or tissue injury, Pro-MSP was cleared from the circulation with a half-life of approximately 100 min, This rapid clearance was not peculiar to I-125-pro- MSP, since clearance rates of unlabeled pro-MSP and of I-125-bovine serum albumin were comparable, The Liver was the major locus of radioactivity 10-20 min after the intravenous injection of I-125-pro-MSP. By 90 min, over 60% of total recovered radioactivity was in the small intestine, Reflecting gastrointestinal transit, counts decreased in the small intestine and appeared in the colon by 180 min, Essentially all counts in urine and feces obtained at later times were soluble in trichloracetic acid, These findings reflected rapid hepatic proteolysis of pro-MSP to fragments undetectable by antibody to pro-MSP; within 20 min after intravenous administration, immuno-precipitable counts were only 22% of the total liver extract radioactivity. Comparison of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and radioautography data for immunoprecipitated plasma and liver extract revealed no evidence for hepatic conversion of pro-MSP to MSP. Thus, the hepatic catabolic pathway of pro-MSP is degradative and does not yield mature MSP, The results support our view that MSP is not released into the circulation but is generated at specific extravascular loci by pro-MSP convertases. RP Leonard, EJ (reprint author), NCI,FREDERICK CANC RES & DEV CTR,IMMUNOPATHOL SECT,IMMUNOBIOL LAB,BLDG 560,RM 12-71,FREDERICK,MD 21702, USA. NR 32 TC 5 Z9 5 U1 0 U2 0 PU FEDERATION AMER SOC EXP BIOL PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 SN 0741-5400 J9 J LEUKOCYTE BIOL JI J. Leukoc. Biol. PD OCT PY 1996 VL 60 IS 4 BP 453 EP 458 PG 6 WC Cell Biology; Hematology; Immunology SC Cell Biology; Hematology; Immunology GA VM935 UT WOS:A1996VM93500003 PM 8864128 ER PT J AU Ortaldo, JR Mason, AT Longo, DL Beckwith, M Creekmore, SP McVicar, DW AF Ortaldo, JR Mason, AT Longo, DL Beckwith, M Creekmore, SP McVicar, DW TI T cell activation via the disialoganglioside GD3: Analysis of signal transduction SO JOURNAL OF LEUKOCYTE BIOLOGY LA English DT Article DE PLC gamma phosphorylation; calcium flux; herbimycin A; R24 monoclonal antibody ID NATURAL-KILLER-CELLS; MONOCLONAL-ANTIBODIES; GANGLIOSIDE GD3; LYMPHOCYTE-RESPONSES; TYROSINE KINASE; RECEPTOR; PHOSPHORYLATION; STIMULATION; MELANOMA; PROTEIN AB The monoclonal antibody (mAb) R24 is a murine immunoglobulin G(3) (IgG(3)) that reacts with the GD3 disialoganglioside present on melanoma cells as well as a subset of T cells. R24 mAb has induced antitumor responses both alone and in combination with interleukin-2 (IL-2) in clinical trials. We have reported T cell activation via GD3 as measured by the induction of tyrosine phosphorylation. In this study a more detailed analysis of signal transduction after ligation of GD3 was performed in an attempt to understand the mechanism of in vivo therapeutic benefits observed, Analysis of subsequent events indicated that GD3 engagement resulted in phospholipase C gamma phosphorylation and calcium flux, When ras-associated events were examined, GD3 signaling resulted in ras activation as determined by GDP/GTP conversion as well as dose- and time-dependent IP3 activation, In addition, the majority of the IP3 activation by GD3 was inhibited by herbimycin A pretreatment. Elucidation of the nature and potential role of this moiety in GD3 signal transduction should be useful. Collectively, these data suggest a novel mechanism of T cell activation via a single, non-protein, surface moiety, This novel form of T cell-mediated activation may permit the delivery and local activation of effector cells at the tumor resulting in site-specific activation of the immune system. C1 NCI,FREDERICK CANC RES & DEV CTR,BIOL RESOURCES BRANCH,FREDERICK,MD 21702. NCI,FREDERICK CANC RES & DEV CTR,OFF ASSOCIATE DIRECTOR,BIOL RESPONSE MODIFIERS PROGRAM,DCT,FREDERICK,MD 21702. NCI,FREDERICK CANC RES & DEV CTR,PRI DYNCORP,BIOL CARCINOGENESIS & DEV PROGRAM,FREDERICK,MD 21702. RP Ortaldo, JR (reprint author), NCI,FREDERICK CANC RES & DEV CTR,DBS,EXPT IMMUNOL LAB,BLDG 560,RM 31-39,FREDERICK,MD 21702, USA. RI McVicar, Daniel/G-1970-2015 NR 26 TC 30 Z9 30 U1 1 U2 1 PU FEDERATION AMER SOC EXP BIOL PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 SN 0741-5400 J9 J LEUKOCYTE BIOL JI J. Leukoc. Biol. PD OCT PY 1996 VL 60 IS 4 BP 533 EP 539 PG 7 WC Cell Biology; Hematology; Immunology SC Cell Biology; Hematology; Immunology GA VM935 UT WOS:A1996VM93500014 PM 8864139 ER PT J AU Sakai, N SantamarinaFojo, S Yamashita, S Matsuzawa, Y Brewer, HB AF Sakai, N SantamarinaFojo, S Yamashita, S Matsuzawa, Y Brewer, HB TI Exon 10 skipping caused by intron 10 splice donor site mutation in cholesteryl ester transfer protein gene results in abnormal downstream splice site selection SO JOURNAL OF LIPID RESEARCH LA English DT Article DE cholesteryl ester transfer protein (CETP); deficiency; hyperalphalipoproteinemia; mutation; splicing; skipping; atherosclerosis; high density lipoprotein (HDL) ID HIGH-DENSITY-LIPOPROTEINS; MESSENGER-RNA; FAMILIAL HYPERALPHALIPOPROTEINEMIA; NONSENSE MUTATIONS; MISSENSE MUTATION; DEFICIENCY; DEFECT; PARTICLES; FREQUENCY; SEQUENCES AB Cholesteryl ester transfer protein (CETP) deficiency is the most common. cause of hyperalphalipoproteinemia in Japan. However, the genetic basis of this disorder has not been fully characterized. We have studied a 49-year-old Japanese male presenting with total cholesterol, HDL cholesterol, and apolipoprotein A-I levels of 300, 236, and 233 mg/dl, respectively, and total absence of CETP activity and mass in plasma. Sequence analysis of the patient's CETP gene revealed that the splice donor consensus GT was substituted by GG in intron 10 (intron 10 splice defect) and by AT in intron 14 (intron 14 splice defect). Restriction digestion of PCR-amplified DNA using NdeI and MaeIII established that the patient was a compound heterozygote for both gene defects. Sequencing of cDNA amplified by RT-PCR from the patient's monocyte-derived macrophage RNA demonstrated abnormal splicing with deletion of exon 10 as well as alternative splicing at a native AG site located 31 nucleotides 5' of the normal splice acceptor in intron 13. Thus, the intron 10 splice defect results in exon 10 skipping and the insertion of a 31 bp fragment between exon 13 and exon 14, which contains an in frame stop codon. The presence of abnormally spliced mRNA. was further confirmed by amplification of patient cDNA using CETP specific primers. Abnormal splicing of exon 14 as a result of the intron 14 splice defect was not detected, indicating potential unstable CETP mRNA derived from that mutation. These findings demonstrate that a novel splice site mutation in intron 10 of the CETP gene results in the skipping of exon 10, as well as disruption of downstream splicing at intron 13 identifying a novel mechanism leading to CETP deficiency. C1 OSAKA UNIV,SCH MED,DEPT INTERNAL MED 2,SUITA,OSAKA 565,JAPAN. RP Sakai, N (reprint author), NHLBI,MOL DIS BRANCH,NIH,10 CTR DR,MSC 1666,BETHESDA,MD 20892, USA. NR 46 TC 40 Z9 41 U1 0 U2 0 PU LIPID RESEARCH INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 SN 0022-2275 J9 J LIPID RES JI J. Lipid Res. PD OCT PY 1996 VL 37 IS 10 BP 2065 EP 2073 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VQ508 UT WOS:A1996VQ50800002 PM 8906584 ER PT J AU Smith, GH AF Smith, Gilbert H. TI TGF-beta and Functional Differentiation SO JOURNAL OF MAMMARY GLAND BIOLOGY AND NEOPLASIA LA English DT Article DE Transforming growth factor beta; lactation; mammary gland AB A review of the pertinent literature suggests that TGF-beta 1 may play a multifaceted role in functional differentiation of mammary epithelium. Evidence for the expression of TGF-beta 1 RNA and the presence of functional TGF-beta 1 protein in differentiating mammary epithelial cells from a pregnant mouse has been recently reported. The specific role of mammary-epithelial-cell-produced TGF-beta 1 in the differentiating mammary gland is presently unclear. However, several possible functions are suggested from the following observations. Milk protein production is negatively regulated by exogenous TGF-beta 1 during gestational development of the gland but not during lactation. Consistent with reports linking TGF-beta 1 gene expression with mammary gland involution following lactation, overexpression of TGF-beta 1 in the differentiating secretory epithelium leads to premature programmed cell death in the absence of a negative effect on secretory epithelial cell proliferation. A role for TGF-beta 1 in cell cycle control and suppression of malignant progression independent from its inhibitory effect on epithelial cell growth has been demonstrated in keratinocytes. A similar function could provide protection against malignancy in proliferating mammary epithelium and account for TGF-beta 1 suppression of mammary tumorigenesis in transgenic mice overexpressing transforming growth factor alpha (TGF-alpha).(3) C1 [Smith, Gilbert H.] NCI, Oncogenet Sect, Lab Tumor Immunol & Biol, Bethesda, MD 20892 USA. RP Smith, GH (reprint author), NIH, Bldg 10,Room 8B07, Bethesda, MD 20892 USA. EM SmithG@ltiblp.nci.nih.gov NR 27 TC 21 Z9 22 U1 0 U2 0 PU SPRINGER/PLENUM PUBLISHERS PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 1083-3021 J9 J MAMMARY GLAND BIOL JI J. Mammary Gland Biol. Neoplasia PD OCT PY 1996 VL 1 IS 4 BP 343 EP 352 DI 10.1007/BF02017390 PG 10 WC Oncology; Endocrinology & Metabolism; Physiology SC Oncology; Endocrinology & Metabolism; Physiology GA V25HE UT WOS:000208468300003 PM 10887508 ER PT J AU Clay, JR AF Clay, JR TI Effects of permeant cations on K+ channel gating in nerve axons revisited SO JOURNAL OF MEMBRANE BIOLOGY LA English DT Article DE K+ channel; ion effects on gating; nerve axon ID SQUID GIANT-AXONS; POTASSIUM CHANNEL; EXTERNAL CESIUM; IONS; RUBIDIUM; KINETICS; LYMPHOCYTES; CURRENTS AB An increase in extracellular potassium ion concentration, K-o, significantly slows the potassium channel deactivation rate in squid giant axons, as previously shown. Surprisingly, the effect does not occur in all preparations which, coupled with the voltage independence of this result in preparations in which it does occur, suggests that it is mediated at a site outside of the electric field of the channel, and that this site is accessible to potassium ions in some preparations, but not in others. In other words, the effect does not appear to be related to occupancy of the channel by potassium ions. This conclusion is supported by a four-barrier, three-binding site model of single file diffusion through the channel in which one site, at most, is unoccupied by a potassium ion (single-vacancy model). The model is consistent with current-voltage relations with various levels of K-o, and, by definition, with multiple occupancy by K+. The model predicts that occupancy of any given site is essentially independent of K-o (or K-i). The effects of extracellular Rb+ and Cs+ on gating are strongly voltage dependent, and they were observed in all preparations investigated. Consequently, the mechanism underlying these results would appear to be different from that which underlies the effect of K+ on gating. In particular, the effect of Rb+ on gating is reduced by strong hyperpolarization, which in the context of the occupancy hypothesis, is consistent with the voltage dependence of the current-voltage relation in the presence of Rb+. The primary, novel, finding in this study is that the effects of Cs+ are counterintuitive in this regard. Specifically, the slowing of channel deactivation rate by Cs+ is also reduced by hyperpolarization, similar to the Rb+ results, whereas blockade is enhanced, which is seemingly inconsistent with the concept that occupancy of the channel by Cs+ underlies the effect of this ion on gating. This result is further elucidated by barrier modeling of the current-voltage relation in the presence of Cs+. C1 MARINE BIOL LAB,WOODS HOLE,MA 02543. RP Clay, JR (reprint author), NINCDS,NEUROPHYSIOL LAB,NIH,BETHESDA,MD 20892, USA. NR 22 TC 4 Z9 4 U1 0 U2 0 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 SN 0022-2631 J9 J MEMBRANE BIOL JI J. Membr. Biol. PD OCT PY 1996 VL 153 IS 3 BP 195 EP 201 PG 7 WC Biochemistry & Molecular Biology; Cell Biology; Physiology SC Biochemistry & Molecular Biology; Cell Biology; Physiology GA VM550 UT WOS:A1996VM55000003 PM 8849414 ER PT J AU Mienville, JM Barker, JL Lange, GD AF Mienville, JM Barker, JL Lange, GD TI Mechanosensitive properties of BK channels from embryonic rat neuroepithelium SO JOURNAL OF MEMBRANE BIOLOGY LA English DT Article DE Ca2+-activated K+ channels; stretch; mechanosensitivity; cytoskeleton; rat neuroepithelium; patch clamp ID ACTIVATED ION CHANNELS; SMOOTH-MUSCLE CELLS; STRETCH ACTIVATION; MEMBRANE STRETCH; SKELETAL-MUSCLE; K+ CHANNELS; FATTY-ACIDS; NEURONS; CURRENTS AB The mechanosensitive properties of large-conductance Ca2+-activated K+ (BK) channels from embryonic rat neuroepithelium were investigated with the cell-attached and inside-out configurations of the patch-clamp technique, The channels were activated in both recording configurations by negative pressures applied to the patch electrode, but reversal of the effect was total and immediate in inside-out patches whereas it was incomplete and delayed in on-cell patches, This mechanosensitivity was not mediated by Ca2+ ions or fatty acids, suggesting that it is an intrinsic property of these channels, Cytochalasin B did not affect mechanosensitivity in on-cell patches but increased it in inside-out patches, Kinetic studies showed that stretch increased the mean open time of the channels and decreased the slowest time constant of their closed-time distributions, The present as well as previous results suggest complex interactions between embryonic BK channels and their membranous and submembranous environment. C1 NINCDS,INSTRUMENTAT & COMP SECT,NIH,BETHESDA,MD 20892. RP Mienville, JM (reprint author), NINCDS,NEUROPHYSIOL LAB,LNP,NIH,BLDG 36,ROOM 2C02 36 CONVENT DR,MSC 4066,BETHESDA,MD 20892, USA. NR 24 TC 29 Z9 31 U1 0 U2 3 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 SN 0022-2631 J9 J MEMBRANE BIOL JI J. Membr. Biol. PD OCT PY 1996 VL 153 IS 3 BP 211 EP 216 DI 10.1007/s002329900124 PG 6 WC Biochemistry & Molecular Biology; Cell Biology; Physiology SC Biochemistry & Molecular Biology; Cell Biology; Physiology GA VM550 UT WOS:A1996VM55000005 PM 8849416 ER PT J AU Goldstein, SR Kidder, LH Herne, TM Levin, IW Lewis, EN AF Goldstein, SR Kidder, LH Herne, TM Levin, IW Lewis, EN TI The design and implementation of a high-fidelity Raman imaging microscope SO JOURNAL OF MICROSCOPY-OXFORD LA English DT Article DE spectroscopic imaging; Raman spectroscopy; acousto-optic tunable filter; chemical imaging ID TUNABLE FILTERS; SPECTROSCOPY; OPERATION; SPECTRUM; DETECTOR; LASER AB We describe a Raman imaging microscope that produces high-fidelity, large format Raman images and Raman spectra from samples as small as 1 mu m in size, Laser illumination is delivered to the object by means of an infinity corrected microscope objective, either by a galvanometer scanning system or a widefield fibre optic, Wavelength selection of Raman scattered emission is achieved by an acousto-optic tunable filter (AOTF), which maintains image fidelity and provides either continuous or random wavelength selection, The collimated AOTF output is imaged first by a tube lens and then by a projection lens onto a cooled silicon CCD array, Instrument features, including factors that determine the system's spatial and spectral resolution, and design considerations are discussed in detail, Images and spectra of test objects and samples that demonstrate the capability of this imaging spectrometer are presented, The potential of intrinsic chemical imaging is discussed in terms of its use in the analyses of a variety of chemical and biological samples. C1 NIDDKD,PHYS CHEM LAB,NIH,BETHESDA,MD 20892. NIH,NATL CTR RES RESOURCES,BIOMED ENGN & INSTRUMENTAT PROGRAM,BETHESDA,MD 20892. NR 24 TC 22 Z9 22 U1 0 U2 5 PU BLACKWELL SCIENCE LTD PI OXFORD PA OSNEY MEAD, OXFORD, OXON, ENGLAND OX2 0EL SN 0022-2720 J9 J MICROSC-OXFORD JI J. Microsc.-Oxf. PD OCT PY 1996 VL 184 BP 35 EP 45 DI 10.1046/j.1365-2818.1996.1130670.x PN 1 PG 11 WC Microscopy SC Microscopy GA VQ860 UT WOS:A1996VQ86000007 PM 8923757 ER PT J AU Wong, M Ramayya, MS Chrousos, GP Driggers, PH Parker, KL AF Wong, M Ramayya, MS Chrousos, GP Driggers, PH Parker, KL TI Cloning and sequence analysis of the human gene encoding steroidogenic factor 1 SO JOURNAL OF MOLECULAR ENDOCRINOLOGY LA English DT Article ID HORMONE-RECEPTOR SUPERFAMILY; VENTROMEDIAL HYPOTHALAMIC NUCLEUS; ADRENAL HYPOPLASIA CONGENITA; REPEAT-BINDING PROTEIN; TRANSCRIPTION FACTOR; AD4-BINDING PROTEIN; GONADAL DEVELOPMENT; ENZYME EXPRESSION; KEY REGULATOR; FTZ-F1 GENE AB The orphan nuclear receptor steroidogenic factor 1 (SF-1) plays key roles in endocrine development and function. Initially identified as a positive regulator of the cytochrome P450 steroid hydroxylases, analyses of knockout mice deficient in SF-1 revealed that SF-1 is essential for adrenal and gonadal development, pituitary gonadotropin expression and formation of the ventromedial hypothalamic nucleus. Although more limited in suggested that SF-1 is important for differentiated function in adrenocortical and gonadotrope adenomas. In the hope of extending our understanding of SF-1 function by identifying possible roles of SF-1 in clinical endocrine disorders, we isolated the FTZ-F1 gene encoding human SF-1 and mapped it to chromosome 9q33. In this report, we characterize the sequence and structural organization of the human cDNA and gene encoding SF-1, providing new insights into comparative aspects of SF-1 structure that will facilitate efforts to study the role of this transcription factor in human endocrine disorders. C1 DUKE UNIV,MED CTR,HOWARD HUGHES MED INST,DURHAM,NC 27710. DUKE UNIV,MED CTR,DEPT MED,DURHAM,NC 27710. DUKE UNIV,MED CTR,DEPT PHARMACOL,DURHAM,NC 27710. NICHHD,NIH,DEV ENDOCRINOL BRANCH,BETHESDA,MD 20892. NR 37 TC 69 Z9 71 U1 0 U2 0 PU J ENDOCRINOLOGY LTD PI BRISTOL PA 17/18 THE COURTYARD, WOODLANDS, ALMONDSBURY, BRISTOL, ENGLAND BS12 4NQ SN 0952-5041 J9 J MOL ENDOCRINOL JI J. Mol. Endocrinol. PD OCT PY 1996 VL 17 IS 2 BP 139 EP 147 DI 10.1677/jme.0.0170139 PG 9 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VT284 UT WOS:A1996VT28400006 PM 8938589 ER PT J AU Sterneck, E Kaplan, DR Johnson, PF AF Sterneck, E Kaplan, DR Johnson, PF TI Interleukin-6 induces expression of peripherin and cooperates with Trk receptor signaling to promote neuronal differentiation in PC12 cells SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE interleukin-6; nerve growth factor; PC12 cells; neurite outgrowth; peripherin; mitogen-activated protein kinase ID NERVE GROWTH-FACTOR; LEUKEMIA INHIBITORY FACTOR; CILIARY NEUROTROPHIC FACTOR; KINASE-ACTIVITY; MESSENGER-RNA; TYROSINE PHOSPHORYLATION; CHOLINERGIC NEURONS; NEURITE OUTGROWTH; GENE-EXPRESSION; RAT-BRAIN AB In contrast to the intensively studied nerve growth factor (NGF)-related family of cytokines, relatively little is known about the mechanisms of neurotrophic activity elicited by the cytokine interleukin-6 (IL-6). We have examined the mechanisms of IL-6-induced neuronal differentiation of the pheochromocytoma cell line PC12. IL-6 independently induced the expression of peripherin, identifying this gene as the first neuronal-specific target of IL-6. However, IL-6 alone failed to elicit neurite outgrowth in PC12 cells and instead required low levels of Trk/NGF receptor tyrosine kinase activity to induce neuronal differentiation. The cooperating Trk signal could be provided by either overexpression of Trk or exposure to low concentrations of NGF. IL-6 also functioned cooperatively with basic fibroblast growth factor to promote PC12 differentiation, IL-6 and Trk/NGF synergized in enhancing tyrosine phosphorylation of the Erk-1 mitogen-activated protein kinase and in activating expression of certain NGF target genes. NGF also induced expression of the gp80 subunit of the IL-6 receptor, providing another potential mechanism of cooperativity between NGF and IL-6 signaling, We propose that IL-6 functions as an enhancer of NGF signaling rather than as an autonomous neuronal differentiation signal. Moreover, our results demonstrate that a Trk receptor-specific cellular response can be achieved in the absence of NGF through amplification of its basal signaling activity by the IL-6 receptor system. C1 NCI,EUKARYOT TRANSCRIPT REGULAT GRP,MBCL,ABL BASIC RES PROGRAM,FREDERICK CANC RES & DEV CTR,FREDERICK,MD 21702. NCI,EUKARYOT SIGNAL TRANSDUCT SECT,MBCL,ABL BASIC RES PROGRAM,FREDERICK CANC RES & DEV CTR,FREDERICK,MD 21702. RI Johnson, Peter/A-1940-2012 OI Johnson, Peter/0000-0002-4145-4725 NR 62 TC 58 Z9 60 U1 0 U2 2 PU LIPPINCOTT-RAVEN PUBL PI PHILADELPHIA PA 227 EAST WASHINGTON SQ, PHILADELPHIA, PA 19106 SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD OCT PY 1996 VL 67 IS 4 BP 1365 EP 1374 PG 10 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA VH578 UT WOS:A1996VH57800005 PM 8858917 ER PT J AU Shetty, HU Smith, QR Washizaki, K Rapoport, SI Purdon, AD AF Shetty, HU Smith, QR Washizaki, K Rapoport, SI Purdon, AD TI Identification of two molecular species of rat brain phosphatidylcholine that rapidly incorporate and turn over arachidonic acid in vivo SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE arachidonic acid; turnover; phospholipids; brain; molecular species; signal transduction; half-life; metabolism ID PERFORMANCE LIQUID-CHROMATOGRAPHY; PLATELET-ACTIVATING FACTOR; STIMULATED HUMAN-PLATELETS; FATTY-ACIDS; UNANESTHETIZED RATS; SIGNAL TRANSDUCTION; QUANTITATIVE METHOD; INOSITOL LIPIDS; MOUSE-BRAIN; PHOSPHOLIPIDS AB In vivo rates of arachidonic acid incorporation and turnover were determined for molecular species of rat brain phosphatidylcholine (PtdCho) and phosphatidylinositol (PtdIns). [H-3]Arachidonic acid was infused intravenously in pentobarbital-anesthetized rats at a programmed rate to maintain constant plasma specific activity for 2-10 min. At the end of infusion, animals were killed by microwave irradiation, and brain phospholipids were isolated, converted to diacylglycerobenzoates, and resolved as molecular species by reversed-phase HPLC. Most [H-3]arachidonate (>87%) was incorporated into PtdCho and PtdIns, with arachidonic acid at the sn-2 position and with oleic acid (18:1), palmitic acid (16:0), or stearic acid (18:0) at the sn-1 position. However, 10-15% of labeled brain PtdCho eluted in a small peak containing two molecular species with arachidonic acid at the sn-2 position and palmitoleic acid (16:1) or linoleic acid (18:2) at the sn-1 position. Analysis demonstrated that tracer was present in both the 16:1-20:4 and 18:2-20:4 PtdCho species at specific activities 10-40 times that of the other phospholipids. Based on the measured mass of arachidonate in each phospholipid molecular species, half-lives were calculated for arachidonate of <10 min in 16:1-20:4 and 18:2-20:4 PtdCho and 1-3 h in 16:0-20:4, 18:0-20:4, and 18:1-20:4 PtdCho and PtdIns. The very short half-lives for arachidonate in the 16:1-20:4 and 18:2-20:4 PtdCho molecular species suggest important roles for these molecules in brain phospholipid metabolism and signal transduction. C1 NIA, NEUROSCI LAB, NIH, BETHESDA, MD 20892 USA. NR 43 TC 33 Z9 33 U1 0 U2 1 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 OCT PY 1996 VL 67 IS 4 BP 1702 EP 1710 PG 9 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA VH578 UT WOS:A1996VH57800044 PM 8858956 ER PT J AU Callahan, P Baumann, MH Rabii, J AF Callahan, P Baumann, MH Rabii, J TI Inhibition of tuberoinfundibular dopaminergic neural activity during suckling: Involvement of mu and kappa opiate receptor subtypes SO JOURNAL OF NEUROENDOCRINOLOGY LA English DT Article DE nor-binaltorphimine; beta-funaltrexamine; lactation ID DELTA OPIOID RECEPTOR; PROLACTIN-RELEASE; RAT-BRAIN; PHARMACOLOGICAL CHARACTERIZATION; BETA-ENDORPHIN; MOLECULAR-CLONING; FEMALE RATS; FUNCTIONAL EXPRESSION; CDNA CLONING; SECRETION AB Previous studies have shown that mu (mu) and kappa (kappa) opioid antagonists inhibit suckling-induced prolactin release, Prolactin responses elicited by pup suckling or opioid administration are mediated, at least in part, by suppression of dopamine (DA) release from tuberoinfundibular dopaminergic (TIDA) neurons in the hypothalamus. We examined the effects of the mu opiate receptor antagonist, beta-funaltrexamine (beta-FNA), and the kappa opiate receptor antagonist, nor-binaltorphimine (nor-BNI) on the activity of TIDA neurons in lactating rats. TIDA neuronal activity was determined by measuring DOPA accumulation in the caudate putamen (CP) and median eminence (ME). The effects of opioid antagonist treatment were determined in pup-deprived (low circulating prolactin levels) or pup-suckled rats (high circulating prolactin levels), The accumulation of 5-hydroxytryptophan (5-HTP) in the medial preoptic area (MPOA), the anterior hypothalamus (AH) and the median eminence (ME) was quantified as an index of serotonergic activity in the same animals for comparative purposes. In vehicle treated rats, suckling caused a significant and selective decrease in DOPA accumulation in the ME, beta-FNA (5 mu g, i.c.v.) pretreatment significantly increased DOPA accumulation in the ME of pup-deprived and pup-suckled rats, beta-FNA pretreatment also prevented the suckling-induced suppression of DOPA accumulation in the ME, In contrast to the actions of beta-FNA, pretreatment with nor-BNI (8 mu g, i.c.v.) did not significantly affect the activity of the TIDA neurons in pup-deprived or pup-suckled rats. Suckling alone did not alter 5-HTP accumulation in any of the brain regions examined, and neither opioid antagonist had appreciable effects on 5-HTP accumulation. These results demonstrate that the EOP tonically inhibit the TIDA neurons in both pup-deprived and pup-suckled, post-partum female rats by acting through the mu, but not the kappa, opiate receptor subtype, Furthermore, the suckling-induced inhibition of TIDA neurons is also mediated through the EOP acting at mu, but not kappa opioid receptors. C1 RUTGERS STATE UNIV,DEPT BIOL SCI,NELSON BIOL LABS,PISCATAWAY,NJ 08855. RUTGERS STATE UNIV,BUR BIOL RES,PISCATAWAY,NJ 08855. NIDA,CLIN PSYCHOPHARMACOL LAB,INTRAMURAL RES PROGRAM,BALTIMORE,MD 21224. MIAMI UNIV,CTR NEUROSCI,DEPT ZOOL,OXFORD,OH 45056. NR 53 TC 39 Z9 39 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA OSNEY MEAD, OXFORD, OXON, ENGLAND OX2 0EL SN 0953-8194 J9 J NEUROENDOCRINOL JI J. Neuroendocrinol. PD OCT PY 1996 VL 8 IS 10 BP 771 EP 776 DI 10.1046/j.1365-2826.1996.05207.x PG 6 WC Endocrinology & Metabolism; Neurosciences SC Endocrinology & Metabolism; Neurosciences & Neurology GA VL126 UT WOS:A1996VL12600007 PM 8910807 ER PT J AU Maity, R Mukherjee, R Skolnick, P AF Maity, R Mukherjee, R Skolnick, P TI Stereoselective inhibition of natural killer activity by the sigma ligand (+)-pentazocine SO JOURNAL OF NEUROIMMUNOLOGY LA English DT Article DE sigma receptors; pentazocine; NK cell activity; splenocyte ID CELL-MEDIATED CYTOTOXICITY; GUINEA-PIG BRAIN; RECEPTORS MODULATE; H-3 (+)-PENTAZOCINE; BINDING-SITES; HIGHLY POTENT; VAS-DEFERENS; IN-VIVO; IDENTIFICATION; PHENCYCLIDINE AB The effect of sigma (sigma) receptor ligands on natural killer (NK) activity was examined both in vivo and in vitro. Following injection of mice with sigma receptor ligands such as (+)-pentazocine, (-)-pentazocine, ED 1073, ED 1165 and ED 737, NK activity was measured in poly-I.C.-stimulated mouse splenocytes. (+)-Pentazocine reduced NK cell activity in a dose-dependent fashion, while the (-) enantiomer was inactive in this measure. For example, at a dose of 50 mg/kg, (+)-pentazocine suppressed NK activity (using effector to target cell ratios of 200:1, 100:1 and 50:1) by > 70%, 24 h after injection while (-)-pentazocine was inactive. The other sigma ligands examined either slightly enhanced or had no effect on NK activity. Nonetheless, parenteral administration of the sigma receptor ligand ED 1165 blocked (+)-pentazocine-induced suppression of NK activity, while the opiate receptor antagonist naltrexone was ineffective. Addition of sigma receptor ligands (10(-11)-10(-5) M) to splenocyte cultures for 24 h did not affect NK activity. These findings indicate that while sigma receptor ligands are capable of modulating NK activity, this effect is not the result of an action on splenocyte sigma receptors, but may be mediated via sigma receptors either in the central or peripheral nervous systems. C1 NIDDK,NEUROSCI LAB,NIH,BETHESDA,MD 20892. NR 59 TC 2 Z9 3 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0165-5728 J9 J NEUROIMMUNOL JI J. Neuroimmunol. PD OCT PY 1996 VL 70 IS 1 BP 7 EP 13 DI 10.1016/S0165-5728(96)00046-X PG 7 WC Immunology; Neurosciences SC Immunology; Neurosciences & Neurology GA VK832 UT WOS:A1996VK83200002 PM 8862129 ER PT J AU Yoshimura, N DeGroat, WC AF Yoshimura, N DeGroat, WC TI Characterization of voltage-sensitive Na+ and K+ currents recorded from acutely dissociated pelvic ganglion neurons of the adult rat SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID CULTURED PARASYMPATHETIC NEURONS; DORSAL-ROOT GANGLIA; CALCIUM CURRENTS; SODIUM CURRENTS; PATCH-CLAMP; IMMUNOHISTOCHEMICAL CHARACTERIZATION; INTRACARDIAC GANGLIA; SYMPATHETIC NEURON; POTASSIUM CURRENTS; AFFERENT NEURONS AB 1. Electrophysiological properties of acutely dissociated neurons from the major pelvic ganglion (MPG) of the adult male rat were studied with whole cell patch-clamp recording techniques. The MPG neurons innervating the urinary bladder were labeled by retrograde axonal tracing methods with the use of a fluorescent dye, Fast Blue (FB), injected into the bladder wall and identified with a fluorescent microscope. 2. Passive and active membrane properties such as resting membrane potential, input resistance, duration of action potentials, thresholds for spike activation, or duration of afterhyperpolarization in unidentified MPG neurons were comparable with those of FB-labeled neurons innervating the urinary bladder. The action potential in both unidentified and bladder efferent MPG neurons was reversibly abolished by tetrodotoxin (TTX, 1 mu M). The after-hyperpolarization of the TTX-sensitive action potential in both groups was reduced by application of Cd2+ (0.1 mM) and further suppressed by tetraethylammonium (TEA, 10 mM). Extracellularly applied TEA increased the duration of the action potential, and 4-aminopyridine (4-AP, 1 or 2 mM) also reduced the spike afterhyperpolarization and increased the spike duration. The duration of the action potential was decreased and the rate of spike repolarization was increased by similar to 2.5-fold with negative shift of membrane potential from -40 to -80 mV. 3. The isolated Na+ current was reversibly blocked by 1 mu M TTX and had a mean peak amplitude of 127.3 pA/pF when activated from a holding potential of -70 mV in the external solution containing 100 mM Na+. The Na+ conductance reached half-maximal activation at a membrane potential of -21.5 mV with a slope factor of 4.9 mV. The steady-state inactivation of Na+ conductance occurred at membrane potentials more depolarized than -90 mV, and the half-maximal inactivation was obtained at -57.5 mV with a slope factor of 8.8 mV. 4. The fast-transient A-type K+ current (I-A) was activated at membrane potentials more depolarized than -60 mV from a holding membrane potential of -100 mV, reached a peak amplitude within 10 ms after the onset of depolarizing voltage steps, and decayed within 20-30 ms at membrane potential depolarizations to +20 to +30 mV. The I-A current activated by a voltage step to +20 mV from a holding potential of -100 mV averaged 102.1 pA/pF. The half-maximal activation of the I-A conductance was obtained at a membrane potential of -21.2 mV with a slope factor of 9.9 mV. In steady-state inactivation of I-A current, the half-maximal inactivation occurred at -76.5 mV and the slope factor was 8.0 mV. 5. The delayed K+ current was reduced by 25-35% by bath application of Cd2+ or the elimination of extracellular Ca2+ ions. The bath application of 4-AP (2 mM) suppressed the I-A current by 75% and the delayed K+ current by 60%. Extracellularly applied TEA (10 mM) suppressed the delayed K+ current by 90%, but suppressed the I-A current by only 16%. 6. These results indicate that bladder neurons and unidentified neurons in the MPG have similar properties including a TTX-sensitive Na+ current and three distinct types of voltage-sensitive K+ currents-I-A current, Ca2+-activating K+ current, and delayed rectifier K+ current-that contribute to the repolarization phase of the action potential. These electrical properties of the MPG neurons resemble those of sympathetic neurons in the superior cervical and inferior mesenteric ganglia. C1 NIAAA,LAB PHYSIOL & PHARMACOL STUDIES,SECT ELECTROPHYSIOL,ROCKVILLE,MD 20892. RP Yoshimura, N (reprint author), UNIV PITTSBURGH,SCH MED,DEPT PHARMACOL,E1303 BIOMED SCI TOWER,PITTSBURGH,PA 15261, USA. NR 34 TC 13 Z9 13 U1 0 U2 2 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-3077 J9 J NEUROPHYSIOL JI J. Neurophysiol. PD OCT PY 1996 VL 76 IS 4 BP 2508 EP 2521 PG 14 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA VN613 UT WOS:A1996VN61300034 PM 8899623 ER PT J AU Li, MX Jia, M Fields, RD Nelson, PG AF Li, MX Jia, M Fields, RD Nelson, PG TI Modulation of calcium currents by electrical activity SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID IMMEDIATE-EARLY GENES; GROWTH CONE MOTILITY; NEURITE ELONGATION; GANGLION-CELLS; MEMBRANE DEPOLARIZATION; INTRACELLULAR CALCIUM; SENSORY NEURONS; CA-2+ CHANNELS; NERVOUS-SYSTEM; C-FOS AB 1. Electrical activation of mouse dorsal root ganglion (DRG) neurons in cultures for 1-2 days produced a downregulation of voltage-sensitive calcium currents, which persisted for greater than or equal to 24 h after stimulation was terminated. This regulation varied with different patterns of activation. Both the magnitude and time course of regulation of the low-threshold voltage-activated (LVA) and high-threshold voltage-activated (HVA) currents were differentially sensitive to neural impulse activity. Tonic stimulation at 0.5 Hz did not affect the HVA currents, but 2.5 Hz did produce a significant decrease. Phasic stimulation (10 Hz for 0.5 s every 2 s) with an average frequency of 2.5 Hz produced significantly more downregulation of HVA currents than did the tonic 2.5-Hz stimulation. The efficacy of phasic stimulation varied inversely with the interval between bursts. Thus phasic stimulation of 10 Hz for 0.5 s but delivered every 4 s produced no effects on HVA currents. Stimulation optimal for downregulation of Ca2+ currents also produced a decreased binding by the DRG neurons of an L-type Ca2+ channel antagonist. This suggests a downregulation by electrical activity of the number of Ca2+ channels, rather than an alteration in a constant number of channels. Depression of LVA currents was produced by all stimulus patterns tested, including 0.5-Hz tonic stimulation. Chronic stimulation with a stimulation pattern that downregulated Ca2+ currents also produced a slowing of the increase in intracellular Ca2+ (as measured by Fura-2/AM) that is produced acutely by repetitive stimulation. This is consonant with earlier studies of intracellular Ca2+ concentration kinetics in growth cones. C1 NICHHD,DEV NEUROBIOL LAB,NIH,BETHESDA,MD 20892. NR 42 TC 35 Z9 35 U1 0 U2 0 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-3077 J9 J NEUROPHYSIOL JI J. Neurophysiol. PD OCT PY 1996 VL 76 IS 4 BP 2595 EP 2607 PG 13 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA VN613 UT WOS:A1996VN61300041 PM 8899630 ER PT J AU Wigdahl, B Brady, JN AF Wigdahl, B Brady, JN TI Molecular aspects of HTLV-I: Relationship to neurological diseases SO JOURNAL OF NEUROVIROLOGY LA English DT Review DE HTLV-I; Tax; TSP/HAM; transformation; transcription ID VIRUS-TYPE-I; T-CELL LEUKEMIA; LONG TERMINAL REPEAT; TROPICAL SPASTIC PARAPARESIS; COLONY-STIMULATING FACTOR; CENTRAL-NERVOUS-SYSTEM; MAJOR HISTOCOMPATIBILITY COMPLEX; TRANSCRIPTIONAL CONTROL REGION; CAMP-RESPONSIVE ELEMENT; BASAL GENE-EXPRESSION C1 PENN STATE UNIV,DEPT MICROBIOL & IMMUNOL,HERSHEY,PA 17033. NCI,MOL VIROL LAB,NIH,BETHESDA,MD 20892. NR 133 TC 7 Z9 7 U1 0 U2 0 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE, HAMPSHIRE, ENGLAND RG21 6XS SN 1355-0284 J9 J NEUROVIROL JI J. Neurovirol. PD OCT PY 1996 VL 2 IS 5 BP 307 EP 322 DI 10.3109/13550289609146895 PG 16 WC Neurosciences; Virology SC Neurosciences & Neurology; Virology GA VR762 UT WOS:A1996VR76200003 PM 8912207 ER PT J AU BruckerDavis, F Reynolds, JC Skarulis, MC Fraker, DL Alexander, HR Weintraub, BD Robbins, J AF BruckerDavis, F Reynolds, JC Skarulis, MC Fraker, DL Alexander, HR Weintraub, BD Robbins, J TI False-positive iodine-131 whole-body scans due to cholecystitis and sebaceous cyst SO JOURNAL OF NUCLEAR MEDICINE LA English DT Article DE whole-body scan; iodine-131; sebaceous cyst; cholecystitis; thyroid cancer ID DIFFERENTIATED THYROID-CANCER; CONCISE COMMUNICATION; PERICARDIAL-EFFUSION; SERUM THYROGLOBULIN; RADIOIODINE UPTAKE; TRACHEOSTOMY SITE; I-131 UPTAKE; CARCINOMA; LOCALIZATION; DIVERTICULUM AB False-positive whole-body I-131 scans are not frequent but have serious consequences in the management of patients with thyroid cancer. They can be classified in four main groups: elimination of iodine in body fluids, infection or inflammation, cysts or transudates and nonthyroid tumors. We report on two patients with false-positive post-therapy I-131 scans. The first patient had uptake projected in the right pelvic area which was later proven to be a large gluteal sebaceous cyst. The second patient had uptake in the gallbladder area that did not disappear after I-131 treatment; she underwent exploratory laparotomy which revealed extensive chronic cholecystitis. These cases illustrate two new causes of false-positive I-131 whole-body scans (sebaceous cyst and cholecystitis), which highlights two mechanisms (elimination in body fluid and inflammation). C1 NIDDKD, GENET BIOCHEM BRANCH, NIH, BETHESDA, MD 20892 USA. NIH, DEPT NUCL MED, BETHESDA, MD 20892 USA. NCI, SURG METAB SECT, SURG BRANCH, NIH, BETHESDA, MD 20892 USA. RP BruckerDavis, F (reprint author), NIDDKD, MOL & CELLULAR ENDOCRINOL BRANCH,NIH,BLDG 10, RM 8D14,10 CENT DR, MSC 1758, BETHESDA, MD 20892 USA. NR 46 TC 41 Z9 41 U1 0 U2 0 PU SOC NUCLEAR MEDICINE INC PI RESTON PA 1850 SAMUEL MORSE DR, RESTON, VA 20190-5316 USA SN 0161-5505 EI 1535-5667 J9 J NUCL MED JI J. Nucl. Med. PD OCT PY 1996 VL 37 IS 10 BP 1690 EP 1693 PG 4 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA VL581 UT WOS:A1996VL58100036 PM 8862313 ER PT J AU Kiesewetter, DO Jagoda, E Carson, RE Herscovitch, P Eckelman, WC AF Kiesewetter, DO Jagoda, E Carson, RE Herscovitch, P Eckelman, WC TI Biodistribution of muscarinic subtype selective agonists of the thiadiazoyl tetrahydropyridine class. SO JOURNAL OF NUCLEAR MEDICINE LA English DT Meeting Abstract C1 NIH,PET DEPT,BETHESDA,MD 20892. RI Carson, Richard/H-3250-2011 OI Carson, Richard/0000-0002-9338-7966 NR 0 TC 0 Z9 0 U1 0 U2 0 PU SOC NUCLEAR MEDICINE INC PI RESTON PA 1850 SAMUEL MORSE DR, RESTON, VA 22090-5316 SN 0161-5505 J9 J NUCL MED JI J. Nucl. Med. PD OCT PY 1996 VL 37 IS 10 BP 1822 EP 1822 PG 2 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA VL581 UT WOS:A1996VL58100067 ER PT J AU Kiesewetter, DO Jagoda, E Carson, RE Endres, CJ Herscovitch, P Eckelman, WC AF Kiesewetter, DO Jagoda, E Carson, RE Endres, CJ Herscovitch, P Eckelman, WC TI Synthesis and biodistribution of the muscarinic receptor antagonists (R,R)- and (R,S)-fluoromethyl QNB. SO JOURNAL OF NUCLEAR MEDICINE LA English DT Meeting Abstract C1 NIH,PET DEPT,BETHESDA,MD 20892. RI Carson, Richard/H-3250-2011 OI Carson, Richard/0000-0002-9338-7966 NR 0 TC 0 Z9 0 U1 0 U2 0 PU SOC NUCLEAR MEDICINE INC PI RESTON PA 1850 SAMUEL MORSE DR, RESTON, VA 22090-5316 SN 0161-5505 J9 J NUCL MED JI J. Nucl. Med. PD OCT PY 1996 VL 37 IS 10 BP 1823 EP 1823 PG 1 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA VL581 UT WOS:A1996VL58100068 ER PT J AU StarkeReed, PE AF StarkeReed, PE TI Aging SO JOURNAL OF NUTRITION LA English DT Editorial Material RP StarkeReed, PE (reprint author), NIA,NIH,BETHESDA,MD 20892, USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER INST NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-3166 J9 J NUTR JI J. Nutr. PD OCT PY 1996 VL 126 IS 10 BP 2593 EP 2594 PG 2 WC Nutrition & Dietetics SC Nutrition & Dietetics GA VN193 UT WOS:A1996VN19300018 PM 8857522 ER PT J AU Brouwers, PIM Decarli, C Heyes, MP Moss, HA Wolters, PL TudorWilliams, G Civitello, LA Pizzo, PA AF Brouwers, PIM Decarli, C Heyes, MP Moss, HA Wolters, PL TudorWilliams, G Civitello, LA Pizzo, PA TI Neurobehavioral manifestations of symptomatic HIV-1 disease in children: Can nutritional factors play a role? SO JOURNAL OF NUTRITION LA English DT Article; Proceedings Paper CT Workshop on Nutrition in Pediatric HIV Infection - Setting the Research Agenda CY SEP 28-29, 1995 CL BETHESDA, MD SP NIH, Off AIDS Res, NICHHD, NIDDKD, NIAID, NIMH, US FDA, Pediat AIDS Fdn, Natl Dairy Council, Sandoz Nutr Corp, Bristol Meyers Squibb Co, Clintec Nutr Co, Abbott Labs, Ross Prod Div, Serono Labs Inc, Amer Inst Nutr DE human; encephalopathy; pediatric HIV-1 disease; brain imaging; neuropsychological test ID HUMAN-IMMUNODEFICIENCY-VIRUS; INFUSION ZIDOVUDINE THERAPY; NERVOUS-SYSTEM; CEREBROSPINAL-FLUID; METABOLIC ALKALOSIS; MENTAL-DEVELOPMENT; TYPE-1 INFECTION; DEFICIENCY; VITAMIN-B12; ENCEPHALOPATHY AB Central nervous system (CNS) abnormalities are significant and frequent complications of human immunodeficiency virus (HIV-1) infection in infants and children. Although the predominant cause of neurological and neuropsychological abnormalities appears to be related to HIV infection of the CNS, other factors including malnutrition may also play a role. We retrospectively evaluated the association of change in body weight with changes in neurocognitive function, ventricular brain ratio, and cerebrospinal quinolinic acid levels in a small cohort of children (n = 15; mean age 6.3 years) with symptomatic HIV-1 disease before and after 6 months of antiretroviral therapy with continuous intravenous infusion of zidovudine (ZVD). Significant increases in weight and neurocognitive function as well as decreases in ventricular brain ratio and cerebrospinal quinolinic acid levels were noted after therapy. Only the relation between increase in weight and decrease in ventricular brain ratio was statistically significant (P < .01); contrary to expectations, an increase in weight seemed to correlate with a decrease in neurocognitive function (NS). Another group of children treated at the same time with oral intermittent ZVD, but otherwise receiving the same care did not show the same magnitude of improvement in neurocognitive function. These results seem to suggest that general supportive and medical care as well as nutritional factors may only play a limited role in the neurocognitive improvements after antiretroviral therapy with continuous infusion ZVD. Our sample size was, however, small and the nutritional measure rather global; thus these findings have to be considered as very preliminary. C1 NINCDS,EPILEPSY RES BRANCH,BETHESDA,MD 20892. NIMH,CLIN SCI LAB,BETHESDA,MD 20892. MED ILLNESS COUNSELING CTR,CHEVY CHASE,MD 20814. CHILDRENS NATL MED CTR,WASHINGTON,DC 20010. RP Brouwers, PIM (reprint author), NCI,PEDIAT BRANCH,NIH,ROOM 13N240,BETHESDA,MD 20892, USA. RI DeCarli, Charles/B-5541-2009 NR 62 TC 7 Z9 7 U1 0 U2 0 PU AMER INST NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-3166 J9 J NUTR JI J. Nutr. PD OCT PY 1996 VL 126 IS 10 SU S BP S2651 EP S2662 PG 12 WC Nutrition & Dietetics SC Nutrition & Dietetics GA VN547 UT WOS:A1996VN54700013 ER PT J AU Fowler, MG Rogers, MF AF Fowler, MG Rogers, MF TI Overview of perinatal HIV infection SO JOURNAL OF NUTRITION LA English DT Article; Proceedings Paper CT Workshop on Nutrition in Pediatric HIV Infection - Setting the Research Agenda CY SEP 28-29, 1995 CL BETHESDA, MD SP NIH, Off AIDS Res, NICHHD, NIDDKD, NIAID, NIMH, US FDA, Pediat AIDS Fdn, Natl Dairy Council, Sandoz Nutr Corp, Bristol Meyers Squibb Co, Clintec Nutr Co, Abbott Labs, Ross Prod Div, Serono Labs Inc, Amer Inst Nutr DE perinatal HIV infection; epidemiology; risk factors ID HUMAN-IMMUNODEFICIENCY-VIRUS; TO-CHILD TRANSMISSION; VERTICAL TRANSMISSION; UNITED-STATES; TYPE-1; MOTHER; WOMEN; PREVALENCE; ANTIBODIES; DELIVERY AB The global HIV epidemic is having a profound impact on the health and survival of children. As of 1994, it is estimated that about 2 million children worldwide (WHO, 1994) and 12 thousand children in the United States are HIV infected (Davis et al, 1995). Almost all HIV infection among infants and young children are from mother-to-infant transmission. By the year 2000, HIV is projected to infect 40 million men, women, and children unless effective prevention strategies are developed. Perinatal HIV transmission rates currently vary from 14-40% with the lowest rates being seen in Europe and highest rates in Africa. Known risk factors for perinatal transmission include advanced maternal HIV disease, lower CD4+ counts, and increased viral burden during pregnancy. Observational cohort data suggest that prenatal vitamin A levels, maternal drug use, and duration of membrane rupture during labor also are related to risk of transmission. The United States clinical trial utilizing an antiretroviral (zidovudine [AZT]) prenatally, intrapartum, and for 6 weeks to the infant demonstrated that perinatal HIV transmission was reduced by two-thirds. This dramatic result gives strong encouragement that simpler perinatal prevention strategies applicable to developing countries may also be successful. A number of international studies are underway or planned including perinatal vitamin A and micronutrient trials in Africa. C1 CTR DIS CONTROL & PREVENT,EPIDEMIOL BRANCH,DIV HIV AIDS,ATLANTA,GA 30333. RP Fowler, MG (reprint author), NIAID,EFFICACY TRIALS BRANCH,DIV AIDS,NIH,2A09 SOLAR BLDG,6003 EXECUT BLVD,ROCKVILLE,MD 20852, USA. NR 51 TC 12 Z9 13 U1 0 U2 0 PU AMER INST NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-3166 J9 J NUTR JI J. Nutr. PD OCT PY 1996 VL 126 IS 10 SU S BP S2602 EP S2607 PG 6 WC Nutrition & Dietetics SC Nutrition & Dietetics GA VN547 UT WOS:A1996VN54700004 ER PT J AU Hamilton, F AF Hamilton, F TI Work group session report: Priorities, recommendations, and strategies for supporting the research agenda SO JOURNAL OF NUTRITION LA English DT Editorial Material RP Hamilton, F (reprint author), NIDDKD,GASTROINTESTINAL PROGRAM,BETHESDA,MD 20892, USA. NR 0 TC 1 Z9 1 U1 0 U2 0 PU AMER INST NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-3166 J9 J NUTR JI J. Nutr. PD OCT PY 1996 VL 126 IS 10 SU S BP S2688 EP S2690 PG 3 WC Nutrition & Dietetics SC Nutrition & Dietetics GA VN547 UT WOS:A1996VN54700019 ER PT J AU Hirschfeld, S AF Hirschfeld, S TI Dysregulation of growth and development in HIV-infected children SO JOURNAL OF NUTRITION LA English DT Article; Proceedings Paper CT Workshop on Nutrition in Pediatric HIV Infection - Setting the Research Agenda CY SEP 28-29, 1995 CL BETHESDA, MD SP NIH, Off AIDS Res, NICHHD, NIDDKD, NIAID, NIMH, US FDA, Pediat AIDS Fdn, Natl Dairy Council, Sandoz Nutr Corp, Bristol Meyers Squibb Co, Clintec Nutr Co, Abbott Labs, Ross Prod Div, Serono Labs Inc, Amer Inst Nutr DE growth; endocrine system; nutrition; metabolism ID IMMUNODEFICIENCY-VIRUS INFECTION; RESTING ENERGY-EXPENDITURE; IGF-BINDING-PROTEINS; DOUBLY LABELED WATER; FACTOR-I; SOMATOMEDIN-C; T-CELL; PROGNOSTIC INDICATOR; HEMOPHILIA GROWTH; BODY-COMPOSITION AB Growth dysregulation is quite common in HIV-infected children and growth failure is one of the most sensitive indicators of disease progression. Beginning at birth, HIV-infected infants often have smaller size and lower birthweight than noninfected children born to HIV-infected women. The causes of growth dysregulation are varied, and can be due to alterations in gastrointestinal function, chronic or repetitive infections, and alterations in metabolic and endocrine function. The metabolic and endocrine effects may be the consequence of the primary infection or secondary to the use of any of the medications required to treat HIV infection and its complications. Correlational studies have identified an inverse relationship between viral burden and linear growth and body mass index, i.e., the use of antiviral medications that reduce viral burden is associated with improvements in anthropometric indices of growth. Alterations in cytokine profiles, possibly related to reported abnormalities in thyroid indices, fat metabolism, and the somatomedin axis, may be indicative of dysregulation on a cellular level. Pubertal delay, especially among boys, is common, and may contribute to the overall growth failure associated with HIV infection, If the basis for growth failure resides in metabolic and regulatory abnormalities, then interventions beyond increasing caloric intake will be necessary to increase linear growth rate and reverse growth failure in HIV-infected children. RP Hirschfeld, S (reprint author), NCI,PEDIAT BRANCH,NIH,BETHESDA,MD 20892, USA. RI Hirschfeld, Steven/E-2987-2016 OI Hirschfeld, Steven/0000-0003-0627-7249 NR 123 TC 12 Z9 12 U1 0 U2 1 PU AMER INST NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-3166 J9 J NUTR JI J. Nutr. PD OCT PY 1996 VL 126 IS 10 SU S BP S2641 EP S2650 PG 10 WC Nutrition & Dietetics SC Nutrition & Dietetics GA VN547 UT WOS:A1996VN54700012 ER PT J AU Hirschfeld, S AF Hirschfeld, S TI Working group session report: Setting the basic research agenda SO JOURNAL OF NUTRITION LA English DT Editorial Material RP Hirschfeld, S (reprint author), NCI,BETHESDA,MD 20892, USA. RI Hirschfeld, Steven/E-2987-2016 OI Hirschfeld, Steven/0000-0003-0627-7249 NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER INST NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-3166 J9 J NUTR JI J. Nutr. PD OCT PY 1996 VL 126 IS 10 SU S BP S2680 EP S2680 PG 1 WC Nutrition & Dietetics SC Nutrition & Dietetics GA VN547 UT WOS:A1996VN54700016 ER PT J AU Paul, W AF Paul, W TI Nutrition in pediatric HIV infection: Setting the research agenda - Keynote address SO JOURNAL OF NUTRITION LA English DT Editorial Material RP Paul, W (reprint author), NIH,OFF AIDS RES,BETHESDA,MD 20892, USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER INST NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-3166 J9 J NUTR JI J. Nutr. PD OCT PY 1996 VL 126 IS 10 SU S BP S2599 EP S2601 PG 3 WC Nutrition & Dietetics SC Nutrition & Dietetics GA VN547 UT WOS:A1996VN54700003 ER PT J AU Solomons, N AdeniyiJones, S AF Solomons, N AdeniyiJones, S TI Working group session report: Coordination of domestic and international effort in executing a research agenda in nutrition and pediatric HIV infection SO JOURNAL OF NUTRITION LA English DT Editorial Material C1 NCI,BETHESDA,MD 20892. RP Solomons, N (reprint author), CESSIAM,GUATEMALA CITY,GUATEMALA. NR 1 TC 0 Z9 0 U1 0 U2 0 PU AMER INST NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-3166 J9 J NUTR JI J. Nutr. PD OCT PY 1996 VL 126 IS 10 SU S BP S2684 EP S2687 PG 4 WC Nutrition & Dietetics SC Nutrition & Dietetics GA VN547 UT WOS:A1996VN54700018 ER PT J AU Nagourney, BA Kramer, MS Klebanoff, MA Usher, RH AF Nagourney, BA Kramer, MS Klebanoff, MA Usher, RH TI Recurrent respiratory distress syndrome in successive preterm pregnancies SO JOURNAL OF PEDIATRICS LA English DT Article ID HYALINE-MEMBRANE DISEASE; FIBROBLAST-PNEUMONOCYTE FACTOR; PREMATURE-INFANTS; BETAMETHASONE; DELIVERY; SEX; PREVENTION AB Background: Earlier studies suggesting an increased recurrence risk of respiratory distress syndrome (RDS) among the subsequent infants of women with a previously affected infant were based on low birth weight inclusion criteria that did not differentiate between preterm and growth-retarded infants. Methods: We therefore carried out two cohort studies of women who delivered two singleton preterm (gestational age <37 completed weeks) infants: 1978 to 1989 at the Royal Victoria Hospital (RVH) in Montreal and 1959 to 1966 in the United States Collaborative Perinatal Project (CPP). We compared the relative risk (RR) of the development of RDS in the second infant according to the RDS status of the first. The diagnosis of RDS was based on respiratory distress of more than 24 hours' duration and a reticulogranular pattern on a chest radiograph. Results: The RVH study sample comprised 284 infants born to 142 women, and the CPP sample 642 infants born to 321 mothers. In the RVH cohort the crude RR of RDS in the second sibling was 3.3 (95% confidence interval = 1.0 to 15.1) in women whose first preterm infant had RDS versus those whose first preterm infant did not have RDS. In the CPP cohort the corresponding RR was 2.5 (95% confidence interval = 0.8 to 7.9). These elevated risks were not altered substantially when multiple logistic regression was used to control for potentially confounding factors known to influence the risk of RDS (gestational age, sex, route of delivery, antenatal corticosteroids, and respiratory depression at birth). Conclusions: We conclude that preterm infants born to women with a previous preterm infant affected by RDS are at an increased risk of RDS, which suggests an important genetic (or other familial) tendency in its origin. C1 MCGILL UNIV, FAC MED, DEPT PEDIAT, MONTREAL, PQ H3A 2T5, CANADA. MCGILL UNIV, FAC MED, DEPT OBSTET & GYNECOL, MONTREAL, PQ H3A 2T5, CANADA. MCGILL UNIV, FAC MED, DEPT EPIDEMIOL & BIOSTAT, MONTREAL, PQ H3A 2T5, CANADA. ROYAL VICTORIA HOSP, MONTREAL, PQ H3A 1A1, CANADA. NICHHD, BETHESDA, MD 20892 USA. NR 38 TC 8 Z9 9 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 OCT PY 1996 VL 129 IS 4 BP 591 EP 596 DI 10.1016/S0022-3476(96)70125-7 PG 6 WC Pediatrics SC Pediatrics GA VP565 UT WOS:A1996VP56500017 PM 8859267 ER PT J AU Albandar, JM Brown, LJ Brunelle, JA Loe, H AF Albandar, JM Brown, LJ Brunelle, JA Loe, H TI Gingival state and dental calculus in early-onset periodontitis SO JOURNAL OF PERIODONTOLOGY LA English DT Article DE periodontal diseases diagnosis; periodontal attachment; gingivitis/diagnosis; dental calculus adverse effects; adolescents; sex factors; race factors; controlled clinical trials ID JUVENILE PERIODONTITIS; DESTRUCTIVE FORMS; YOUNG-ADULTS; DISEASE; PREVALENCE; ADOLESCENTS; NIGERIA; LAGOS AB THIS STUDY WAS UNDERTAKEN TO 1) compare the prevalence of gingival inflammation and dental calculus in adolescents with early-onset periodontitis and their matched controls and 2) assess and compare the relationship between the presence of dental calculus and the extent of gingival bleeding and attachment loss in these subjects. The study group consisted of 1,285 13 to 20 year-old individuals, 651 males and 634 females, selected from a national survey of the oral health of U.S. adolescents in 1986/1987. It included 709 (55.2%) Blacks, 224 (17.4%) Hispanics, and 352 (27.4%) Whites. Eighty-nine subjects had localized or generalized juvenile periodontitis (JP), 218 had incidental attachment loss (IAL), and 978 were without clinical attachment loss (controls). The controls were matched to cases on gender, race, age, and geographic location. The subjects were examined clinically to assess the percentage of sites with gingival bleeding and supragingival calculus only and subgingival calculus with or without supragingival calculus. The IAL and JP groups had significantly more gingival bleeding and subgingival calculus than the controls. Also, the JP group had significantly higher prevalence of both conditions than the IAL group. Th percentage of sites with supragingival calculus was not different between the groups, but varied by ethnicity. Hispanics with JP had the highest percentage of sites with gingival bleeding and subgingival calculus, and the lowest percentage of sites with only supragingival calculus. The results demonstrate that gingival inflammation and subgingival calculus are associated with early periodontal breakdown, and contradict earlier reports of early-onset periodontitis not being associated with these factors. RP Albandar, JM (reprint author), NIDR,DIV EPIDEMIOL & ORAL DIS PREVENT,NATCHER BLDG,4AS-13K,45 CTR DR,BETHESDA,MD 20892, USA. OI Albandar, Jasim M./0000-0001-7801-3811 NR 20 TC 37 Z9 38 U1 0 U2 1 PU AMER ACAD PERIODONTOLOGY PI CHICAGO PA 737 NORTH MICHIGAN AVENUE, SUITE 800, CHICAGO, IL 60611-2690 SN 0022-3492 J9 J PERIODONTOL JI J. Periodont. PD OCT PY 1996 VL 67 IS 10 BP 953 EP 959 PG 7 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA VP282 UT WOS:A1996VP28200001 PM 8910833 ER PT J AU Albandar, JM Brown, LJ Loe, H AF Albandar, JM Brown, LJ Loe, H TI Dental caries and tooth loss in adolescents with early-onset periodontitis SO JOURNAL OF PERIODONTOLOGY LA English DT Article DE adolescents; controlled clinical trials; dental caries epidemiology; tooth loss epidemiology; periodontitis, early-onset complications; follow-up studies ID JUVENILE PERIODONTITIS; UNITED-STATES; PROGRESSION; DISEASES; REASONS; ADULTS AB THE AIM OF THIS STUDY WAS TO ASSESS the caries experience and tooth loss over 6 years in subjects with early-onset periodontitis as compared to their matched controls, and to describe the characteristics of teeth lost during this period. A multi-stage probability sample representing 8th to 12th grade U.S. schoolchildren were screened during the 1986/1987 school year to identify subjects with early-onset periodontitis (cases). The examination included measuring the clinical attachment level, presence of caries and dental restorations, and tooth loss. A random sample of controls without early-onset periodontitis were selected for a follow-up examination and were matched to cases on gender, race, age, and geographic location. A total of 266 subjects, with a mean age of 16 years at baseline, were examined during the 1992/1993 school year and were classified into localized (LJP) and generalized juvenile periodontitis (GJP), incidental attachment loss (IAL), and control groups. Whites had more caries experience than Blacks and Hispanics, but there were no significant differences in tooth loss between the ethnic groups. The LJP and the IAL groups, respectively, had higher and lower overall caries experience than the control group. The LJP group had a significantly higher number of missing teeth at follow-up, and exhibited more extensive tooth mortality during 6 years than the control group. The GJP group also showed more tooth loss than the control group, but the difference was not statistically significant. In the LJP, GJP, IAL, and control groups, respectively, 43%, 32%, 26%, and 18% of the subjects lost teeth over 6 years due to disease. The findings showed differences in caries activity between the early-onset periodontitis groups and a variation by race. The findings suggest that loss of periodontal support was the principal cause for tooth loss in the LJP and GJP groups, and that dental caries was the principal cause for tooth extraction in the IAL and the control groups. RP Albandar, JM (reprint author), NIDR,DIV EPIDEMIOL & ORAL DIS PREVENT,NATCHER BLDG 4AS-13K,45 CTR CR,BETHESDA,MD 20892, USA. OI Albandar, Jasim M./0000-0001-7801-3811 NR 18 TC 20 Z9 21 U1 0 U2 0 PU AMER ACAD PERIODONTOLOGY PI CHICAGO PA 737 NORTH MICHIGAN AVENUE, SUITE 800, CHICAGO, IL 60611-2690 SN 0022-3492 J9 J PERIODONTOL JI J. Periodont. PD OCT PY 1996 VL 67 IS 10 BP 960 EP 967 PG 8 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA VP282 UT WOS:A1996VP28200002 PM 8910834 ER PT J AU Brown, LJ Albandar, JM Brunelle, JA Loe, H AF Brown, LJ Albandar, JM Brunelle, JA Loe, H TI Early-onset periodontitis: Progression of attachment loss during 6 years SO JOURNAL OF PERIODONTOLOGY LA English DT Article DE adolescents; follow-up studies; periodontitis, early-onset; periodontal ID JUVENILE PERIODONTITIS; AGE; DISEASE AB WE STUDIED THE PATTERN OF PROGRESSION of early-onset periodontitis and the change in the extent and severity of the periodontal condition in adolescents who were followed for 6 years. In a national survey of the oral health of U.S. children, 14,013 adolescents were examined clinically in 1986/1987 to assess the periodontal attachment loss of teeth. Individuals with early-onset periodontitis within this population were identified and classified into localized juvenile periodontitis (LJP), generalized juvenile periodontitis (GJP), and incidental attachment loss (IAL) groups. Ninety-one subjects, 13 to 20 years old at baseline, were examined 6 years later. They included 51 males and 40 females; and 72 Blacks, 6 Hispanics, and 13 Whites. They were clinically re-examined and then reclassified according to their periodontal status at follow-up. The severity and extent of these diseases continued to increase during the study period. In teeth that were affected at baseline, the lesions had progressed to include deeper portions of the periodontium, and more of the teeth unaffected at baseline exhibited periodontal attachment loss at follow-up, thus changing the disease characteristics and the basis for the clinical classification. Of the individuals classified with LJP at baseline, 62% continued to have LJP 6 years later and 35% developed GJP. Of those classified with GJP initially, all but two (82%) continued to have GJP at follow-up. Among the IAL group, 28% of subjects developed LJP or GJP, and 30% were reclassified in the no attachment loss group. Molars and incisors were the teeth most often, affected in all three groups. The mean change in attachment loss over 6 years in the LJP, GJP, and IAL groups was 0.45, 1.12, and 0.13 mm, respectively. The present findings demonstrate the limitations of the currently used morphological criteria in the classification of early-onset periodontitis. The findings also suggest that the difference between LJP and GJP is in the number and type of teeth involved, and that the two classifications progress similarly, with some cases of LJP developing into GJP. C1 NIDR,DIV EPIDEMIOL & ORAL DIS PREVENT,BETHESDA,MD 20892. OI Albandar, Jasim M./0000-0001-7801-3811 NR 9 TC 46 Z9 49 U1 2 U2 2 PU AMER ACAD PERIODONTOLOGY PI CHICAGO PA 737 NORTH MICHIGAN AVENUE, SUITE 800, CHICAGO, IL 60611-2690 SN 0022-3492 J9 J PERIODONTOL JI J. Periodont. PD OCT PY 1996 VL 67 IS 10 BP 968 EP 975 PG 8 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA VP282 UT WOS:A1996VP28200003 PM 8910835 ER PT J AU Taylor, GW Burt, BA Becker, MP Genco, RJ Shlossman, M Knowler, WC Pettit, DJ AF Taylor, GW Burt, BA Becker, MP Genco, RJ Shlossman, M Knowler, WC Pettit, DJ TI Severe periodontitis and risk for poor glycemic control in patients with non-insulin-dependent diabetes mellitus SO JOURNAL OF PERIODONTOLOGY LA English DT Article DE periodontitis complications; diabetes mellitus, non-insulin dependent; hyperglycemia; hypoglycemia; longitudinal studies; epidemiology; models, statistical ID PIMA-INDIANS; ACTINOBACILLUS-ACTINOMYCETEMCOMITANS; BACTEROIDES-GINGIVALIS; GLYCATED HEMOGLOBIN; DISEASE; POPULATION; INFECTION; MICROBIOLOGY; INFLAMMATION; DESTRUCTION AB THIS STUDY TESTED THE HYPOTHESIS THAT SEVERE PERIODONTITIS in persons with noninsulin-dependent diabetes mellitus (NIDDM) increases the risk of poor glycemic control. Data from the longitudinal study of residents of the Gila River Indian Community were analyzed for dentate subjects aged 18 to 67, comprising all those: 1) diagnosed at baseline with NIDDM (at least 200 mg/dL plasma glucose after a 2-hour oral glucose tolerance test); 2) with baseline glycosylated hemoglobin (HbA(1)) less than 9%; and 3) who remained dentate during the 2-year follow-up period. Medical and dental examinations were conducted at 2-year intervals, Severe periodontitis was specified two ways for separate analyses: 1) as baseline periodontal attachment loss of 6 mm or more on at least one index tooth; and 2) baseline radiographic bone loss of 50% or more on at least one tooth, Clinical data for loss of periodontal attachment were available for 80 subjects who had at least one follow-up examination, 9 of whom had two follow-up examinations at 2-year intervals after baseline. Radiographic bone loss data were available for 88 subjects who had at least one follow-up examination, 17 of whom had two follow-up examinations. Poor glycemic control was specified as the presence of HbA, of 9% or more at follow-up. To increase the sample size, observations from baseline to second examination and from second to third examinations were combined, To control for non-independence of observations, generalized estimating equations (GEE) were used for regression modeling. Severe periodontitis at baseline was associated with increased risk of poor glycemic control at follow-up. Other statistically significant covariates in the GEE models were: 1) baseline age; 2) level of glycemic control at baseline; 3) having more severe NIDDM at baseline; 4) duration of NIDDM; and 5) smoking at baseline, These results support considering severe periodontitis as a risk factor for poor glycemic control and suggest that physicians treating patients with NIDDM should be alert to the signs of severe periodontitis in managing NIDDM. C1 UNIV MICHIGAN, SCH PUBL HLTH, ANN ARBOR, MI 48109 USA. SUNY BUFFALO, SCH DENT MED, DEPT ORAL BIOL, BUFFALO, NY USA. NIDDK, PHOENIX, AZ USA. RP Taylor, GW (reprint author), UNIV MICHIGAN, SCH DENT, DEPT CARIOL RESTORAT SCI & ENDODONT, 1011 N UNIV, ANN ARBOR, MI 48109 USA. FU NIDCR NIH HHS [DE06514, DE07157] NR 76 TC 237 Z9 250 U1 5 U2 29 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 OCT PY 1996 VL 67 IS 10 SU S BP 1085 EP 1093 PG 9 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA VP849 UT WOS:A1996VP84900007 PM 8910827 ER PT J AU Owens, IS Ritter, JK Yeatman, MT Chen, F AF Owens, IS Ritter, JK Yeatman, MT Chen, F TI The novel UGT1 gene complex links bilirubin, xenobiotics, and therapeutic drug metabolism by encoding UDP-glucuronosyltransferase isozymes with a common carboxyl terminus SO JOURNAL OF PHARMACOKINETICS AND BIOPHARMACEUTICS LA English DT Article DE bilirubin UDP-glucuronosyltransferases; genetic locus; genetic defects; therapeutic drugs; xenobiotics ID HYODEOXYCHOLIC ACID; EXPRESSION; PHENOL; CLONING; GLUCURONIDATION; SEQUENCE; LOCUS; CDNAS; CELLS AB The UDP-glucuronosyltransferase system (transferase) plays an important role in the pharmacokinetics of clearance of endogenous metabolites, therapeutic drugs, and xenobiotics. The human bilirubin and phenol transferases are encoded by the same gene complex which we designate UGT1. The gene arrangement indicates there are 6 exon Is each with a promoter and each of which can predictably undergo differential splicing to the 4 common exons (2 through 5) to generate possibly 6 different mRNAs. The entire unique amino acid terminus of each isoform is encoded by an exon I, and the common carboxyl terminus is encoded by the 4 common exons. Evidence supports the existence of other exon Is upstream of the currently described locus. The 13-bp deletion in exon 2 represents the most common defect, to date, in the Crigler-Najjar, Type I individuals. Different point mutations in the 4 common exons and in exon I of UGT1A, however, also account for defective bilirubin transferase activity. The gene arrangement, in conjunction with the toxicity data from the Gunn rat, leads to the prediction that detoxification of bilirubin, xenobiotics, and therapeutic drugs is linked to the UGT1 locus. The Crigler-Najjar syndromes are uncommon, but the Gilbert individuals are commonly represented in 6% of the population. It is expected that similar to the deleterious mutations in the common region of the UGT1 locus in Crigler-Najjar, Type I individuals, there is a range of moderate to intermediate deleterious mutations in this region of the gene of at least some Gilbert's individuals. Linkages, therefore, at this locus could signal that these individuals are at risk for certain drug toxicities and/or idiosyncratic drug reactions. RP Owens, IS (reprint author), NICHHD,HERITABLE DISORDERS BRANCH,NIH,BETHESDA,MD 20892, USA. NR 16 TC 20 Z9 20 U1 0 U2 0 PU PLENUM PUBL CORP PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 SN 0090-466X J9 J PHARMACOKINET BIOP JI J. Pharmacokinet. Biopharm. PD OCT PY 1996 VL 24 IS 5 BP 491 EP 508 DI 10.1007/BF02353476 PG 18 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA WR148 UT WOS:A1996WR14800004 PM 9131487 ER PT J AU Fleckenstein, AE Pogun, S Carroll, FI Kuhar, MJ AF Fleckenstein, AE Pogun, S Carroll, FI Kuhar, MJ TI Recovery of dopamine transporter binding and function after intrastriatal administration of the irreversible inhibitor RTI-76 {3 beta-(3p-chlorophenyl)tropan-2 beta-carboxylic acid p-isothiocyanatophenylethyl ester hydrochloride} SO JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS LA English DT Article ID PARKINSONS-DISEASE; UPTAKE SITES; H-3 COCAINE; D2-DOPAMINE RECEPTORS; NONHUMAN-PRIMATES; NUCLEUS-ACCUMBENS; CAUDATE-PUTAMEN; RAT STRIATUM; MAZINDOL; METAPHIT AB Effects of in vivo, intrastriatal administration of RTI-76 {3 beta-(3-p-chlorophenyl)tropan-2 beta-carboxylic acid p-isothiocyanatophenylethyl ester hydrochloride}, an irreversible inhibitor of dopamine transporter (DAT) binding in vitro, on [I-125]RTI-55 {3 beta[4-iodophenyl]tropan-2 beta-carboxylic acid methyl ester tartrate} binding to striatal DAT in vitro were examined in male rats. Effects on [H-3]DAT and D-1, dopamine receptor binding in vitro after intrastriatal RTI-76 injection were also determined. One hour after direct intrastriatal injection, RTI-76 caused a dose-related increase in K-D for [I-125]RTI-55 binding in vitro in striatal tissue, without affecting transporter maximum binding (B-max). In contrast, 24 hr after administration, RTI-76 caused a dose-related decrease in striatal DAT B-max without affecting K-D, a decrease that reversed over the next several days. Transport of [H-3]dopamine into synaptosomes was decreased similarly. Intrastriatal injection of reversible inhibitors of DAT, such as cocaine or WIN-35428 {3 beta-[4-fluorophenyl]tropan-2 beta-carboxylic acid methyl ester tartrate}, was without effect on transporter binding 1 and 6 days after administration. RTI-76 had little effect on [H-3]SCH-23390 {R-(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro- 1H-3-benzazepine} binding 1 or 24 hr after intrastriatal injection, indicating at least some selectivity of RTI-76 for DAT. The RTI-76-induced decrease in B-max as well as the concurrent decrease in [H-3]DAT, were reversible,with the T-1/2 of transporter recovery estimated to be 6 days. C1 NATL INST DRUG ABUSE,INTRAMURAL RES PROGRAM,NATL INST HLTH,BALTIMORE,MD. EGE UNIV,CTR BRAIN RES,BORNOVA,IZMIR,TURKEY. EGE UNIV,SCH MED,DEPT PHYSIOL,BORNOVA,IZMIR,TURKEY. RES TRIANGLE INST,RES TRIANGLE PK,NC 27709. EMORY UNIV,DIV NEUROSCI,YERKES REG PRIMATE RES CTR,ATLANTA,GA 30322. RI Pogun, Sakire/A-5816-2010 NR 39 TC 46 Z9 46 U1 1 U2 3 PU WILLIAMS & WILKINS PI BALTIMORE PA 351 WEST CAMDEN ST, BALTIMORE, MD 21201-2436 SN 0022-3565 J9 J PHARMACOL EXP THER JI J. Pharmacol. Exp. Ther. PD OCT PY 1996 VL 279 IS 1 BP 200 EP 206 PG 7 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA VL701 UT WOS:A1996VL70100028 PM 8858994 ER PT J AU Nomeir, AA Markham, P Burka, LT Griffin, RJ Ghanayem, BI AF Nomeir, AA Markham, P Burka, LT Griffin, RJ Ghanayem, BI TI Species differences in the disposition and metabolism of nalidixic acid SO JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS LA English DT Article ID CARCINOGENICITY AB Nalidixic acid (NA) is an antimicrobial drug that has been used to treat urinary tract infections. A study of NA by the National Toxicology Program indicated that chronic administration in the diet at doses equivalent to 82 and 175 mg/kg/day for rats, and 175 and 475 mg/kg/day for mice resulted in neoplastic lesions in the preputial and clitoral glands of male and female Fischer 344 rats, respectively, but not in male and female B6C3F1 mice. Our study was designed to evaluate the metabolic basis of this species and tissue-dependent carcinogenicity. [C-14]NA was administered by oral gavage as a suspension in corn oil at doses of 20, 200 or 500 mg/kg. Based on urinary excretion data, at least 35 to 46 and 57 to 79% of dose was absorbed from the gastrointestinal tracts of mice and rats, respectively. NA-derived radioactivity was excreted primarily in urine and feces. The urinary and fecal metabolite profiles were species dependent. At 72 hr after administration, in both genders of rats and mice, the highest concentrations of radioactivity were observed in the liver, and the lowest were in the brain and adipose tissue. The preputial and clitoral glands of male and female rats, respectively, contained consistently and substantially higher concentrations of radioactivity compared to all other tissues, with the exception of liver. In mice, the levels of radioactivity in these tissues were near or below quantifiable levels. The metabolism and disposition characteristics of NA were linear in male and female mice over a dose range of 20 to 500 mg/kg; in rats, they were dose dependent. Results of this study suggest that the species- and tissue-dependent differences in carcinogenicity of NA were associated with differences in metabolism and disposition between the two species. C1 ARTHUR D LITTLE INC,CAMBRIDGE,MA 02140. NIEHS,RES TRIANGLE PK,NC 27709. FU NIEHS NIH HHS [N01-ES-66138] NR 21 TC 1 Z9 1 U1 0 U2 1 PU WILLIAMS & WILKINS PI BALTIMORE PA 351 WEST CAMDEN ST, BALTIMORE, MD 21201-2436 SN 0022-3565 J9 J PHARMACOL EXP THER JI J. Pharmacol. Exp. Ther. PD OCT PY 1996 VL 279 IS 1 BP 222 EP 230 PG 9 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA VL701 UT WOS:A1996VL70100031 PM 8858997 ER PT J AU Church, PJ Stanley, EF AF Church, PJ Stanley, EF TI Single L-type calcium channel conductance with physiological levels of calcium in chick ciliary ganglion neurons SO JOURNAL OF PHYSIOLOGY-LONDON LA English DT Article ID SURFACE-CHARGE; ION PERMEATION; CA2+ CHANNELS; CELLS; SELECTIVITY; MEMBRANE; CURRENTS; DETERMINANTS; SATURATION; DOMAINS AB 1. Single L-type calcium channels in chick ciliary ganglion neurons were studied at high current resolution in cell-attached patch recordings using quartz-glass micropipettes. 2. A single open-channel current amplitude was observed when Ba2+ was the charge carrier with a conductance of 26 pS at 110 mM barium. However, with 110 mM calcium two current fluctuation amplitudes were observed. These were termed low and high fluctuation amplitudes, Ca-L and Ca-H, and had conductances of 8.8 and 12 pS, respectively. These two levels probably reflect two different channel species. Ca-L was identified as an L-type calcium channel on the basis of resistance to inactivation, conductance, and dihydropyridine sensitivity. 3. Single-channel current fluctuations could be detected with calcium concentrations as low as 1.0 mM. Although the unitary conductance (gamma) was much greater with barium than calcium at every concentration tested, the concentration dependence of conductance was similar for gamma(Ba), gamma(CaH) and gamma(CaL). Fitting the concentration dependencies of these conductances with a Langmuir isotherm gave K-D estimates of 4.7, 5.6 and 5.0 mM for barium, Ca-L and Ca-H, respectively. 4. The single-channel conductance of the L-type channel (gamma(L)) can be described by the relation: conductance (in pS) = 9.2/(1 + 5.6/[Ca]) where [Ca] is the external calcium concentration in the 1.0-110 mM range. Thus, at a physiological external calcium concentration of 2 mM the conductance is 2.4 pS. 5. Ca2+ transport through the L-type calcium channel is particularly sensitive to changes in external calcium concentration in the physiological range but approaches saturation at about 10 mM. This characteristic may optimize the responsiveness of the cell to small changes in ambient calcium concentrations while limiting excess entry in the presence of abnormally high calcium levels. C1 NINCDS, SYNAPT MECH SECT, NIH, BETHESDA, MD 20892 USA. NR 26 TC 67 Z9 70 U1 0 U2 1 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 SN 0022-3751 J9 J PHYSIOL-LONDON JI J. Physiol.-London PD OCT 1 PY 1996 VL 496 IS 1 BP 59 EP 68 PG 10 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA VM287 UT WOS:A1996VM28700006 PM 8910196 ER PT J AU Morrell, CH GordonSalant, S Pearson, JD Brant, LJ Fozard, JL AF Morrell, CH GordonSalant, S Pearson, JD Brant, LJ Fozard, JL TI Age- and gender-specific reference ranges for hearing level and longitudinal changes in hearing level SO JOURNAL OF THE ACOUSTICAL SOCIETY OF AMERICA LA English DT Review AB This paper presents age-specific reference ranges for hearing level and change in hearing level for men and women at 500, 1000, 2000, and 4000 Hz. The percentiles are constructed from data obtained from persons in the Baltimore Longitudinal Study of Aging who were rigorously screened for otological disorders and evidence of noise-induced hearing loss. The resulting percentile curves represent norms for changes in hearing level in the absence of any known otologic disease. These percentile curves provide a reference for detecting when a person deviates from a normal pattern of change, thus helping in diagnosing problems with hearing or in monitoring hearing in occupational settings. The smoothed means and standard deviations of the hearing levels were used to construct the longitudinal percentiles. The percentiles for cross-sectional change were constructed using the skew normal distribution to allow for the percentiles to be asymmetric on either side of the median level. These percentiles are the first reference curves that (1) provide standards for hearing level changes over periods of up to 15 years, (2) account for age differences in the distribution of hearing levels, and (3) are based on data from persons who have been systematically screened for otological disorders and evidence of noise-induced hearing loss. (C) 1996 Acoustical Society of America. C1 NIA,GERONTOL RES CTR,LONGITUDINAL STUDIES BRANCH,BALTIMORE,MD 21224. RP Morrell, CH (reprint author), LOYOLA COLL,DEPT MATH SCI,4501 N CHARLES ST,BALTIMORE,MD 21210, USA. RI Fozard, James Leonard/B-3660-2009 FU NIDCD NIH HHS [1 R03 DC02566-01] NR 14 TC 98 Z9 107 U1 1 U2 8 PU AMER INST PHYSICS PI WOODBURY PA CIRCULATION FULFILLMENT DIV, 500 SUNNYSIDE BLVD, WOODBURY, NY 11797-2999 SN 0001-4966 J9 J ACOUST SOC AM JI J. Acoust. Soc. Am. PD OCT PY 1996 VL 100 IS 4 BP 1949 EP 1967 DI 10.1121/1.417906 PN 1 PG 19 WC Acoustics; Audiology & Speech-Language Pathology SC Acoustics; Audiology & Speech-Language Pathology GA VN885 UT WOS:A1996VN88500004 PM 8865630 ER PT J AU Greenhill, LL Abikoff, HB Arnold, LE Cantwell, DP Conners, CK Elliott, G Hechtman, L Hinshaw, SP Hoza, B Jensen, PS March, JS Newcorn, J Pelham, WE Severe, JB Swanson, JM Vitiello, B Wells, K AF Greenhill, LL Abikoff, HB Arnold, LE Cantwell, DP Conners, CK Elliott, G Hechtman, L Hinshaw, SP Hoza, B Jensen, PS March, JS Newcorn, J Pelham, WE Severe, JB Swanson, JM Vitiello, B Wells, K TI Medication treatment strategies in the MTA study: Relevance to clinicians and researchers SO JOURNAL OF THE AMERICAN ACADEMY OF CHILD AND ADOLESCENT PSYCHIATRY LA English DT Article DE attention-deficit hyperactivity disorder; multisite trial; methylphenidate; dextroamphetamine; pemoline; imipramine ID DEFICIT HYPERACTIVITY DISORDER; SUSTAINED-RELEASE METHYLPHENIDATE; PSYCHOTHERAPY-RESEARCH; HYPERKINETIC-CHILDREN; DRUG RESPONSE; DEXTROAMPHETAMINE; PSYCHIATRY; ADOLESCENTS; STIMULANTS; ADHD AB Objective: Clinicians have difficulty applying drug research findings to clinical practice, because research protocols use methods different from those used in daily office practice settings. Method: To design a medication protocol for a multisite clinical trial involving 576 children with attention-deficit hyperactivity disorder (ADHD) while maintaining relevance to clinical practice, investigators from the NIMH Collaborative Multisite Multimodal Treatment Study of Children with Attention-Deficit/Hyperactivity Disorder (MTA study) developed novel medication strategies. These were designed to work either in a monomodal or multimodal format and to ensure standard approaches are used across diverse sites. Each child randomized to medication (projected N = 288) is individually titrated to his or her ''best'' methylphenidate dose and has individual ADHD symptoms monitored. Decision rules were developed to guide ''best dose'' selection, dose changes, medication changes, the management of side effects, and integration with psychosocial treatments. Conclusions: The MTA study uses a controlled method to standardize the identification of each child's ''best'' methylphenidate dose in a national, multisite cooperative treatment program. Although the titration protocol is complex, the study's individual dosing approach and algorithms for openly managing ADHD children's medication over time will be of interest to clinicians in office practice. C1 COLUMBIA UNIV,NEW YORK,NY 10027. NIMH,BETHESDA,MD 20892. LONG ISL JEWISH MED CTR,GLEN OAKS,NY 11004. UNIV CALIF IRVINE,IRVINE,CA 92717. DUKE UNIV,DURHAM,NC 27706. UNIV CALIF BERKELEY,BERKELEY,CA 94720. MCGILL UNIV,MONTREAL,PQ H3A 2T5,CANADA. MT SINAI MED CTR,NEW YORK,NY 10029. UNIV PITTSBURGH,PITTSBURGH,PA 15260. OI Jensen, Peter/0000-0003-2387-0650; Newcorn, Jeffrey /0000-0001-8993-9337 FU NIMH NIH HHS [MH50467, U01-MH50453, UO1 MH50447, U01-MH50461, UO1 MH50461, UO1 MH50440] NR 47 TC 152 Z9 157 U1 1 U2 6 PU WILLIAMS & WILKINS PI BALTIMORE PA 351 WEST CAMDEN ST, BALTIMORE, MD 21201-2436 SN 0890-8567 J9 J AM ACAD CHILD PSY JI J. Am. Acad. Child Adolesc. Psychiatr. PD OCT PY 1996 VL 35 IS 10 BP 1304 EP 1313 DI 10.1097/00004583-199610000-00017 PG 10 WC Psychology, Developmental; Pediatrics; Psychiatry SC Psychology; Pediatrics; Psychiatry GA VK055 UT WOS:A1996VK05500017 PM 8885584 ER PT J AU Slavkin, HC AF Slavkin, HC TI What we know about pain SO JOURNAL OF THE AMERICAN DENTAL ASSOCIATION LA English DT Article RP Slavkin, HC (reprint author), NIDR,31 CTR DR,MSC 2290,BLDG 31,ROOM 2C39,BETHESDA,MD 20892, USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER DENTAL ASSN PI CHICAGO PA 211 E CHICAGO AVE, CHICAGO, IL 60611 SN 0002-8177 J9 J AM DENT ASSOC JI J. Am. Dent. Assoc. PD OCT PY 1996 VL 127 IS 10 BP 1536 EP 1541 PG 6 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA VL780 UT WOS:A1996VL78000022 PM 8908927 ER PT J AU Levey, AS Beck, GJ Bosch, JP Caggiula, AW Greene, T Hunsicker, LG Klahr, S AF Levey, AS Beck, GJ Bosch, JP Caggiula, AW Greene, T Hunsicker, LG Klahr, S TI Short-term effects of protein intake, blood pressure, and antihypertensive therapy on glomerular filtration rate in the modification of diet in renal disease study SO JOURNAL OF THE AMERICAN SOCIETY OF NEPHROLOGY LA English DT Article DE GFR; dietary protein; blood pressure; antihypertensive therapy ID CONVERTING ENZYME-INHIBITORS; DIABETIC NEPHROPATHY; PROGRESSION; INSUFFICIENCY; CAPTOPRIL; ENALAPRIL; TRIAL AB Glomerular filtration rate is often used to assess the level of renal function and the progression of renal disease. However, the short-term effects of dietary protein restriction, blood pressure reduction, and specific classes of antihypertensive agents on GFR may be opposite in direction from their observed longterm beneficial effects on the progression of renal disease, The purpose of these analyses was to characterize these short-term effects and determine whether they can obscure the relationship between renal structure and function in patients with slowly progressive renal disease. The Modification of Diet in Renal Disease Study was a randomized trial of the effect of dietary protein restriction and strict blood pressure control on the decline in GFR in 840 patients with mean (range) baseline GFR of 36.1 (13 to 55) ml/min per 1.73 m(2), In this study, comparisons of the randomized groups and correlational analyses were used to determine the short-term (4-month) effects on GFR of changes in protein intake, mean arterial pressure (MAP), and class of antihypertensive agents (computed as the reduction in GFR associated with starting versus stopping medications) during the first 4 months of follow-up and in subsequent 4-month intervals during the first 2 yr of follow-up. Combining results over the first 2 yr of follow-up, and controlling for changes in antihypertensive medications, the independent effect on GFR of changes in protein intake and MAP was 1.1 ml/min per 0.4 g/kg per day and 0.9 ml/min per 10 mm Hg, respectively (P < 0.001), These effects were observed in patients with increasing or decreasing protein intake or MAP, and in patients with stable or changing antihypertensive regimens. Starting treatment with diuretics, beta-blockers, or angiotensin-converting enzyme inhibitors was associated with a 4.4-, 3.2-, or 2.2-mL/min greater GFR decline, respectively, than was stopping this treatment (P < 0.001). The effect of changes in protein intake, MAP, and diuretics was greater in patients with higher initial GFR. After controlling for initial GFR, there were no significant differences between the short-term effects observed during the first 4 months of follow-up and the short-term effects during subsequent followup. Changes in protein intake, blood pressure, and antihypertensive agents have small but statistically significant short-term effects on GFR. These effects can lead to clinically significant changes in renal function in patients undergoing multiple interventions and are large enough to confound the results of clinical trials in patients with slowly progressive renal disease. Future studies using GFR to assess the progression of renal disease should take into account these shortterm effects when the length of follow-up is being planned. C1 NIDDKD,NIH,BETHESDA,MD. RP Levey, AS (reprint author), TUFTS UNIV NEW ENGLAND MED CTR,DIV NEPHROL,750 WASHINGTON ST,BOSTON,MA 02111, USA. NR 50 TC 44 Z9 47 U1 1 U2 5 PU WILLIAMS & WILKINS PI BALTIMORE PA 351 WEST CAMDEN ST, BALTIMORE, MD 21201-2436 SN 1046-6673 J9 J AM SOC NEPHROL JI J. Am. Soc. Nephrol. PD OCT PY 1996 VL 7 IS 10 BP 2097 EP 2109 PG 13 WC Urology & Nephrology SC Urology & Nephrology GA VQ177 UT WOS:A1996VQ17700008 ER PT J AU Scrimgeour, EM Chand, PR Kenny, K Brown, P AF Scrimgeour, EM Chand, PR Kenny, K Brown, P TI Creutzfeldt-Jakob disease in Oman: Report of two cases SO JOURNAL OF THE NEUROLOGICAL SCIENCES LA English DT Article DE Creutzfeldt-Jakob disease; Oman-Arab race; cerebrospinal fluid ID CEREBROSPINAL-FLUID; DIAGNOSIS AB Sporadic Creutzfeldt-Jakob disease (CJD) was diagnosed in two Omani Arab men, aged 50 and 75 years respectively, both with a history of rapidly developing dementia and myoclonic jerks. Illness developed over a period of 3 months in the first case and over six months in the second. Electroencephalography in both subjects showed periodic triphasic sharp waves characteristic of CJD. In neither case was it possible to obtain a brain biopsy or perform autopsy (autopsy is contrary to Islamic practice in the Middle East), however, electrophoresis of cerebrospinal fluid from the second patient revealed the distinctive double protein spots characteristic of CJD. This is the first report of CJD from Oman. C1 SULTAN QABOOS UNIV,DEPT MED,NEUROL UNIT,AL KHOUD,MUSCAT,OMAN. NIH,CENT NERVOUS SYST STUDIES LAB,BETHESDA,MD 20892. RP Scrimgeour, EM (reprint author), SULTAN QABOOS UNIV,DEPT MED,INFECT DIS UNIT,POB 35,AL KHOUD,MUSCAT,OMAN. NR 5 TC 3 Z9 3 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0022-510X J9 J NEUROL SCI JI J. Neurol. Sci. PD OCT PY 1996 VL 142 IS 1-2 BP 148 EP 150 DI 10.1016/0022-510X(96)00171-2 PG 3 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA VM925 UT WOS:A1996VM92500025 PM 8902735 ER PT J AU Kolobow, T Wang, J Giacomini, M RealiForster, C AF Kolobow, T Wang, J Giacomini, M RealiForster, C TI An adjustable, percutaneously placed helical coil: Control of left ventricular filling pressure during cardiopulmonary bypass in severe left ventricular failure SO JOURNAL OF THORACIC AND CARDIOVASCULAR SURGERY LA English DT Article RP Kolobow, T (reprint author), NHLBI,PULM CRIT CARE MED BRANCH,NIH,BLDG 10,BETHESDA,MD 20892, USA. NR 2 TC 0 Z9 0 U1 0 U2 0 PU MOSBY-YEAR BOOK INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 SN 0022-5223 J9 J THORAC CARDIOV SUR JI J. Thorac. Cardiovasc. Surg. PD OCT PY 1996 VL 112 IS 4 BP 1113 EP 1115 DI 10.1016/S0022-5223(96)70116-2 PG 3 WC Cardiac & Cardiovascular Systems; Respiratory System; Surgery SC Cardiovascular System & Cardiology; Respiratory System; Surgery GA VW275 UT WOS:A1996VW27500034 PM 8873742 ER PT J AU Bukreyev, A Camargo, E Collins, PL AF Bukreyev, A Camargo, E Collins, PL TI Recovery of infectious respiratory syncytial virus expressing an additional, foreign gene SO JOURNAL OF VIROLOGY LA English DT Article ID INFLUENZA-A VIRUS; MAMMALIAN-CELLS; MESSENGER-RNA; REPORTER GENE; CLONED CDNA; GENOMIC RNA; PROTEIN; RESCUE; DNA; TRANSCRIPTION AB A previous report described the recovery from cDNA of infectious recombinant respiratory syncytial virus (RSV) strain A2 (P. L. Collins, M. G. Hill, E. Camargo, H. Grosfeld, R. M. Chanock, and B. R. Murphy, Proc. Natl. Acad. Sci, USA, 92:11563-11567, 1995). Here, the system was used to construct recombinant RSV containing an additional gene encoding chloramphenicol acetyltransferase (CAT). The CAT coding sequence was flanked by RSV-specific gene-start and gene-end motifs; the transcription signals for the viral RNA-dependent RNA polymerase. The RSV-CAT chimeric transcription cassette,vas inserted into the region between the G and F genes of the complete cDNA-encoded positive sense RSV antigenome, and infectious CAT-expressing recombinant RSV was recovered. Transcription of the inserted gene into the predicted subgenomic polyadenylated mRNA was demonstrated by Northern (RNA) blot hybridization analysis, and the encoded protein was detected by enzyme assay and by radioimmunoprecipitation. Quantitation of intracellular CAT, SH, G, and F mRNAs showed that the CAT mRNA was efficiently expressed and that the levels of the G and F mRNAs (which represent the genes on either side of the inserted CAT gene) were comparable to those expressed by a wild-type recombinant RSV. Consistent with this finding, the CAT-containing acid mild-type viruses were very similar with regard to the levels of synthesis of the major viral proteins, Each of 25 RSV isolates obtained by plaque purification following eight serial passages expressed CAT, shoeing that the foreign gene was faithfully maintained in functional form. Analysis by reverse transcription and PCR did not reveal evidence of deletion of the foreign sequence. This finding demonstrated that the RSV genome can accept and maintain an increase in length of 762 nucleotides of foreign sequence and can be engineered to encode an additional, 11th mRNA. The presence of the additional gene resulted in a 10% decrease in plaque diameter and was associated with delay in virus growth and 20-fold decrease in virus yield in vitro. Thus, introduction of an additional gene into the RSV genome might represent a method of attenuation. The ability to express foreign genes by recombinant RSV has implications for basic studies as well as for the development of live recombinant vaccines. C1 NIAID,LID,BETHESDA,MD 20892. NR 41 TC 67 Z9 69 U1 1 U2 3 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0022-538X J9 J VIROL JI J. Virol. PD OCT PY 1996 VL 70 IS 10 BP 6634 EP 6641 PG 8 WC Virology SC Virology GA VG127 UT WOS:A1996VG12700012 PM 8794298 ER PT J AU Carpenter, MA Brown, EW Culver, M Johnson, WE PeconSlattery, J Brousset, D OBrien, SJ AF Carpenter, MA Brown, EW Culver, M Johnson, WE PeconSlattery, J Brousset, D OBrien, SJ TI Genetic and phylogenetic divergence of feline immunodeficiency virus in the puma (Puma concolor) SO JOURNAL OF VIROLOGY LA English DT Article ID NUCLEOTIDE-SEQUENCE ANALYSIS; GENOMIC ORGANIZATION; NONDOMESTIC FELIDS; FLORIDA PANTHER; INFECTION; CATS; LENTIVIRUS; SUBTYPES; SUPERINFECTION; SUBSTITUTIONS AB Feline immunodeficiency virus (FIV) is a lentivirus which causes an AIDS-like disease in domestic cats (Felis catus), A number of other felid species, including the puma (Puma concolor), carry a virus closely related to domestic cat FIV. Serological testing revealed the presence of antibodies to FIV in 22% of 434 samples from throughout the geographic range of the puma. FIV-Pco pal gene sequences isolated from pumas revealed extensive sequence diversity, greater than has been documented in the domestic cat. The puma sequences formed two highly divergent groups, analogous to the clades which have been defined for domestic cat and lion (Panthera lee) FIV. The puma clade A was made up of samples from Florida and California, whereas clade B consisted of samples from other parts of North America, Central America, and Brazil, The difference between these two groups was as great as that reported among three lion FIV clades, Within puma clades, sequence variation is large, comparable to between-clade differences seen for domestic cat clades, allowing recognition of 15 phylogenetic lineages (subclades) among puma FIV-Pco, Large sequence divergence among isolates, nearly complete species monophyly, and widespread geographic distribution suggest that FIV-Pco has evolved within the puma species for a long period. The sequence data provided evidence for vertical transmission of FIV-Pco from mothers to their kittens, for coinfection of individuals by two different viral strains, and for cross-species transmission of FIV from a domestic cat to a puma, These factors may all be important for understanding the epidemiology and natural history of FIV in the puma. C1 NCI,FREDERICK CANC RES & DEV CTR,VIRAL CARCINOGENESIS LAB,FREDERICK,MD 21702. NCI,FREDERICK CANC RES & DEV CTR,SAIC FREDERICK,INTRAMURAL RES SUPPORT PROGRAM,FREDERICK,MD 21702. UNIV NACL AUTONOMA MEXICO,FAC MED VET & ZOOTECNIA,COYOACAN,MEXICO. RI Johnson, Warren/D-4149-2016; OI Johnson, Warren/0000-0002-5954-186X; Carpenter, Margaret/0000-0003-1606-1986 NR 45 TC 76 Z9 76 U1 0 U2 11 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0022-538X J9 J VIROL JI J. Virol. PD OCT PY 1996 VL 70 IS 10 BP 6682 EP 6693 PG 12 WC Virology SC Virology GA VG127 UT WOS:A1996VG12700018 PM 8794304 ER PT J AU Bertran, J Miller, JL Yang, YP FenimoreJustman, A Rueda, F Vanin, EF Nienhuis, AW AF Bertran, J Miller, JL Yang, YP FenimoreJustman, A Rueda, F Vanin, EF Nienhuis, AW TI Recombinant adeno-associated virus-mediated high-efficiency, transient expression of the murine cationic amino acid transporter (Ecotropic retroviral receptor) permits stable transduction of human HeLa cells by ecotropic retroviral vectors SO JOURNAL OF VIROLOGY LA English DT Article ID HEMATOPOIETIC PROGENITOR CELLS; BONE-MARROW TRANSPLANTATION; SITE-SPECIFIC INTEGRATION; UMBILICAL-CORD BLOOD; GAMMA-GLOBIN GENE; ADENOASSOCIATED VIRUS; MAMMALIAN-CELLS; PHENOTYPIC CORRECTION; NONDIVIDING CELLS; DNA INTEGRATION AB Adeno-associated virus has a broad host range, is nonpathogenic, and integrates into a preferred location on chromosome 19, features that have fostered development of recombinant adeno-associated viruses (rAAV) as gene transfer vectors for therapeutic applications. We have used an rAAV to transfer and express the murine cationic amino acid transporter which functions as the ecotropic retroviral receptor, thereby rendering human cells conditionally susceptible to infection by an ecotropic retroviral vector, The proportion of human HeLa cells expressing the receptor at 60 h varied as a function of the multiplicity of infection (MOI) with the rAAV, Cells expressing the ecotropic receptor were efficiently transduced with an ecotropic retroviral vector encoding a nucleus-localized form of beta-galactosidase. Cells coexpressing the ecotropic receptor and nucleus-localized beta-galactosidase were isolated by fluorescence-activated cell sorting, and cell lines were recovered by cloning at limiting dilution. After growth in culture, all clones contained the retroviral vector genome, but fewer than 10% (3 of 47) contained the rAAV genome and continued to express the ecotropic receptor. The ecotropic receptor coding sequences in the rAAV genome were under the control of a tetracycline-modulated promoter. In the presence of tetracycline, receptor expression was low and the proportion of cells transduced by the ecotropic retroviral vector was decreased, Modulation of receptor expression was achieved with both an episomal and an integrated form of the rAAV genome. These data establish that functional gene expression from an rAAV genome can occur transiently without genome integration. C1 ST JUDE CHILDRENS RES HOSP, DEPT HEMATOL ONCOL, DIV EXPT HEMATOL, MEMPHIS, TN 38105 USA. NIDDKD, BIOL CHEM LAB, BETHESDA, MD 20892 USA. GENET THERAPY INC, GAITHERSBURG, MD USA. UNIV TENNESSEE, SCH MED, DEPT MED, MEMPHIS, TN 38163 USA. UNIV TENNESSEE, SCH MED, DEPT PEDIAT, MEMPHIS, TN 38163 USA. RI Rueda, Felix/L-1580-2013 FU NCI NIH HHS [P30CA21765]; NHLBI NIH HHS [P01HL 53749-02] NR 67 TC 35 Z9 35 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD OCT PY 1996 VL 70 IS 10 BP 6759 EP 6766 PG 8 WC Virology SC Virology GA VG127 UT WOS:A1996VG12700027 PM 8794313 ER PT J AU Kovacs, GR Moss, B AF Kovacs, GR Moss, B TI The vaccinia virus H5R gene encodes late gene transcription factor 4: Purification, cloning, and overexpression SO JOURNAL OF VIROLOGY LA English DT Article ID DEPENDENT RNA-POLYMERASE; CAPPING ENZYME; POLY(A) POLYMERASE; VIRAL REPLICATION; INITIATION-FACTOR; SOLUBLE EXTRACT; IDENTIFICATION; VIRIONS; PROTEIN; SUBUNIT AB The vaccinia virus late stage-specific transcription factor P3 was purified to homogeneity from HeLa cells that were infected in the presence of an inhibitor of viral DNA replication. The purified 36-kDa protein was digested with trypsin, and the peptides were analyzed by mass spectroscopy and amino-terminal sequencing. The purified factor was identified as the product of the vaccinia virus H5R open reading frame by both methods. A recombinant baculovirus was engineered to express the H5R open reading frame. The partially purified recombinant protein could replace the vaccinia virus P3 factor in transcription assays. On the basis of these findings, we assigned the H5R gene product the name viral late gene transcription factor 4 (VLTF-4). Unlike VLTF-1, -2, and -3, which are synthesized exclusively after viral DNA replication, VLTF-4 is synthesized before and after viral DNA synthesis. Indirect immunofluorescence of infected cells with anti-H5R protein antiserum demonstrated that VLTF-4 is diffusely distributed in the cytoplasm when DNA replication is blocked but is localized to discrete viral DNA-containing factories during a productive infection. Its expression pattern and subcellular distribution suggest that the H5R gene product may have multiple roles In the viral life cycle. C1 NIAID, VIRAL DIS LAB, NIH, BETHESDA, MD 20892 USA. NR 53 TC 66 Z9 68 U1 1 U2 2 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0022-538X J9 J VIROL JI J. Virol. PD OCT PY 1996 VL 70 IS 10 BP 6796 EP 6802 PG 7 WC Virology SC Virology GA VG127 UT WOS:A1996VG12700032 PM 8794318 ER PT J AU Wickham, TJ Segal, DM Roelvink, PW Carrion, ME Lizonova, A Lee, GM Kovesdi, I AF Wickham, TJ Segal, DM Roelvink, PW Carrion, ME Lizonova, A Lee, GM Kovesdi, I TI Targeted adenovirus gene transfer to endothelial and smooth muscle cells by using bispecific antibodies SO JOURNAL OF VIROLOGY LA English DT Article ID INTEGRIN ALPHA(V)BETA(3); VONWILLEBRAND-FACTOR; CORONARY RESTENOSIS; ADHESION RECEPTOR; MEDIATED TRANSFER; CYSTIC-FIBROSIS; ATTACHMENT; VITRONECTIN; EXPRESSION; INTEGRIN-ALPHA-V-BETA-5 AB A major hurdle to adenovirus (Ad)-mediated gene transfer is that the target tissue lacks sufficient levels of receptors to mediate vector attachment via its fiber coat protein. Endothelial and smooth muscle cells are primary targets in gene therapy approaches to prevent restenosis following angioplasty or to promote or inhibit angiogenesis. However, Ad poorly binds and transduces these cells because of their low or undetectable levels of functional Ad fiber receptor. The Ad-binding deficiency of these cells was overcome by targeting Ad binding to alpha(v) integrin receptors that are sufficiently expressed by these cells. In order to target alpha(v) integrins, a bispecific antibody (bsAb) that comprised a monoclonal Ab to the FLAG peptide epitope, DYKDDDDK, and a monoclonal Ab to alpha(v) integrins was constructed, In conjunction with the bsAb, a nerv vector, AdFLAG, which incorporated the FLAG peptide epitope into its penton base protein was constructed, Complexing AdFLAG with the bsAb increased the beta-glucuronidase transduction of human venule endothelial cells and human intestinal smooth muscle cells by seven- to ninefold compared with transduction by AdFLAG alone. The increased transduction efficiency was shown to occur through the specific interaction of the complex with alpha(v) integrins, These results demonstrate that bsAbs can be successfully used to target Ad to a specific cellular receptor and thereby increase the efficiency of gene transfer. C1 NCI,BETHESDA,MD 20892. RP Wickham, TJ (reprint author), GENVEC INC,12111 PARKTOWN DR,ROCKVILLE,MD 20852, USA. NR 36 TC 238 Z9 243 U1 0 U2 4 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0022-538X J9 J VIROL JI J. Virol. PD OCT PY 1996 VL 70 IS 10 BP 6831 EP 6838 PG 8 WC Virology SC Virology GA VG127 UT WOS:A1996VG12700038 PM 8794324 ER PT J AU Kuo, L Grosfeld, H Cristina, J Hill, MG Collins, PL AF Kuo, L Grosfeld, H Cristina, J Hill, MG Collins, PL TI Effect of mutations in the gene-start and gene-end sequence motifs on transcription of monocistronic and dicistronic minigenomes of respiratory syncytial virus SO JOURNAL OF VIROLOGY LA English DT Article ID VESICULAR STOMATITIS-VIRUS; NUCLEOTIDE-SEQUENCES; MESSENGER-RNAS; SENDAI VIRUS; REPLICATION; GENOME; PROTEIN; INVIVO; ORDER AB Preceding and following each gene of respiratory syncytial virus (RSV) are two conserved sequences, the gene-start (GS) and gene-end (GE) motifs, respectively, which are thought to be transcription signals. The functions and boundaries of these signals and the process of sequential transcription were analyzed with cDNA-encoded RNA analogs (minigenomes) of nonsegmented negative-sense RSV genomic RNA, Two minigenomes were used, The monocistronic RSV-CAT minigenome consists of the chloramphenicol acetyltransferase (CAT) translational open reading frame (ORF) bordered by the GS and GE motifs and flanked by the 3' leader and 5' trailer extragenic regions of genomic RNA, The dicistronic RSV-CAT-LUC minigenome is a derivative of RSV-CAT into which the ORF for luciferase (LUG), bordered by GS and GE motifs, was inserted downstream of the CAT gene with an intergenic region positioned between the two genes, Each minigenome, was synthesized in vitro and transfected into RSV-infected cells, where it was replicated and transcribed to yield the predicted polyadenylated subgenomic mRNA(s), The only RSV sequences required for efficient transcription and RNA replication were the 44-nucleotide 3' leader region, the last 40 nucleotides of the 5' trailer region, and the 9- to 10-nucleotide GS and 12- to 13-nucleotide GE motifs. The GS and GE motifs functioned as self-contained, transportable transcription signals which could be attached to foreign sequences to direct their transcription into subgenomic mRNAs, Removal of the GS motif greatly reduced transcription of its gene, and the requirement for this element was particularly strict for the gene in the downstream position, Ablation of the promoter-proximal GS signal was not associated with increased antigenome synthesis. Consistent with its proposed role in termination and polyadenylation, removal of the CAT GE signal in RSV-CAT resulted in the synthesis of a nonpolyadenylated CAT mRNA, and in RSV-CAT-LUC the same mutation resulted in readthrough transcription to yield a dicistronic CAT-LUG mRNA. The latter result showed that a downstream GS signal is not recognized for reinitiation by the polymerase if it is already engaged in mRNA synthesis; instead, it is recognized only if the polymerase first terminates transcription at an upstream termination signal, This result also showed that ongoing transcription did not open the downstream LUC gene for internal polymerase entry, Removal of both the GS and GE signals of the upstream CAT gene in RSV-CAT-LUC silenced expression of both genes, confirming that independent polymerase entry at an internal gene is insignificant, Remarkably, whereas both genes were silent when the CAT GS and GE signals were both absent, restoration of the CAT CE signal alone restored a significant level (approximately 10 to 12% of the wild-type level) of synthesis of both subgenomic mRNAs. This analysis identified a component of sequential transcription that,vas independent of the promoter-proximal GS signal and appeared to involve readthrough from the leader region. C1 NIAID,INFECT DIS LAB,BETHESDA,MD 20892. NR 41 TC 70 Z9 73 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0022-538X J9 J VIROL JI J. Virol. PD OCT PY 1996 VL 70 IS 10 BP 6892 EP 6901 PG 10 WC Virology SC Virology GA VG127 UT WOS:A1996VG12700046 PM 8794332 ER PT J AU Schneiderman, RD Farrell, KB Wilson, CA Eiden, MV AF Schneiderman, RD Farrell, KB Wilson, CA Eiden, MV TI The Japanese feral mouse Pit1 and Pit2 homologs lack an acidic residue at position 550 but still function as gibbon ape leukemia virus receptors: Implications for virus binding motif SO JOURNAL OF VIROLOGY LA English DT Article ID CELLULAR RECEPTOR; HAMSTER-CELLS; MURINE; RETROVIRUS; INFECTION; GLVR1 AB Murine cells are typically resistant to gibbon ape leukemia virus (GALV). MMMol, a Japanese feral mouse cell line, is an exception in that these cells are susceptible to infection by GALV. We show here that MMMol cells are further distinguished by their unusual receptor properties. MMMol cells infected by GALV are resistant to subsequent infection not only by GALV but also by amphotropic murine leukemia virus. This suggests that GALV can enter MMMol via not only the GALV receptor (MolPit1) but also the amphotropic murine leukemia virus receptor (MolPit2). Therefore, MolPit2 was cloned, sequenced, and compared with the previously reported sequence of MolPit1. Earlier studies have shown that a stretch of nine residues (position 550 to 558) in the fourth extracellular domain of Pit1 is crucial for GALV entry acid that an acidic residue at position 550 is indispensable. However, MolPit1 has isoleucine at this position and MolPit2 has glutamine at the corresponding position (position 522), thus breaking this consensus. To determine what effect these specific changes in the fourth extracellular domain of MolPit1 and MolPit2 have on GALV receptor function, chimeric receptors were made by substituting the fourth extracellular domain of either MolPit1 or MolPit2 for the same region of Pit2, a nonfunctional receptor for GALV. These chimeras were then tested in MDTF, a cell line that lacks functional GALV receptors and is resistant to GALV. Results show that MDTF expressing these chimeras became susceptible to GALV, whereas cells expressing wild-type Pit2 remained resistant. Further, the MolPit1 chimera was identical to Pit1 in efficiency, but the MolPit2 chimera proved substantially less efficient. C1 NIMH,UNIT MOL VIROL,CELL BIOL LAB,BETHESDA,MD 20892. US FDA,DIV CELLULAR & GENE THERAPIES,CTR BIOL EVALUAT & RES,ROCKVILLE,MD 20892. NR 20 TC 15 Z9 15 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0022-538X J9 J VIROL JI J. Virol. PD OCT PY 1996 VL 70 IS 10 BP 6982 EP 6986 PG 5 WC Virology SC Virology GA VG127 UT WOS:A1996VG12700056 PM 8794342 ER PT J AU Yoo, YD Chiou, CJ Choi, KS Yi, YS Michelson, S Kim, S Hayward, GS Kim, SJ AF Yoo, YD Chiou, CJ Choi, KS Yi, YS Michelson, S Kim, S Hayward, GS Kim, SJ TI The IE2 regulatory protein of human cytomegalovirus induces expression of the human transforming growth factor beta 1 gene through an Egr-1 binding site SO JOURNAL OF VIROLOGY LA English DT Article ID IMMEDIATE-EARLY REGION-2; NUCLEAR-LOCALIZATION SIGNAL; CIS-ACTING ELEMENTS; VIRUS EARLY GENE; TRANSCRIPTIONAL REGULATION; RETINOBLASTOMA PROTEIN; FUNCTIONAL INTERACTION; FACTOR-BETA-1 GENE; TRANS-ACTIVATION; TARGET SEQUENCE AB Increases in transforming growth factor beta 1 (TGF-beta 1) mRNA and biological activity in the early phase of human cytomegalovirus (CMV) infection in fibroblasts are paralleled by increased TGF-beta 1-chloramphenicol acetyltransferase (CAT) reporter gene activity. To determine how CMV infection transactivates the TGF-beta 1 promoter, we examined the effects of the cotransfected IE2 regulatory protein of human CMV on 5'-deleted TGF-P beta promoter-CAT reporter genes in transient DNA transfection assays. Two upstream TGF-beta 1 promoter regions each containing an Egr-1 consensus site were shown to he important for IE2-induced transactivation in a cell type that displayed greatly reduced nonspecific activity. Furthermore, transfer of an Egr-1 site from between positions -125 and -98, but not point mutant versions of this site, to a heterologous promoter also conveyed IE2 responsiveness. Addition of an IE2 expression vector or use of the U373 A45 astrocytoma cell line expressing IE2 also produced synergistic stimulation of GAL4-Egr-1-mediated activation of a target promoter containing GAL4 binding sites. The 80-kDa IE2 protein present in A45 cells proved to selectively bind to glutathione S-transferase (GST)-Egr-1 beads. The results of in vitro protein binding assays also revealed that an intact in vitro-translated IE2 protein bound directly to the GST-Egr-1 fusion protein through the zinc finger domain of the Egr-1 protein and that this binding activity was abolished by deletion of parts of the zinc finger DNA-binding domain, Similarly, the Egr-1 protein was found to associate preferentially with a small region within the C-terminal half of the IE2 protein adjacent to the DNA-binding and dimerization domains that are important for both transactivation and downregulation. We conclude from these observations that IE2 may regulate transcription of the TGF-beta 1 gene as well as other potential cellular targets by virtue of its ability to interact with the Egr-1 DNA-binding protein. C1 NCI,CHEMOPREVENT LAB,BETHESDA,MD 20892. JOHNS HOPKINS UNIV,SCH MED,BALTIMORE,MD 21205. SEOUL NATL UNIV,INST MOL BIOL & GENET,SEOUL,SOUTH KOREA. INST PASTEUR,UNITE IMMUNOL VIRALE,F-75724 PARIS 15,FRANCE. FU NIAID NIH HHS [R01 AI24576] NR 68 TC 73 Z9 75 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0022-538X J9 J VIROL JI J. Virol. PD OCT PY 1996 VL 70 IS 10 BP 7062 EP 7070 PG 9 WC Virology SC Virology GA VG127 UT WOS:A1996VG12700065 PM 8794351 ER PT J AU Yu, MS Miller, RH Emerson, S Purcell, RH AF Yu, MS Miller, RH Emerson, S Purcell, RH TI A hydrophobic heptad repeat of the core protein of woodchuck hepatitis virus is required for capsid assembly SO JOURNAL OF VIROLOGY LA English DT Article ID ARGININE-RICH DOMAIN; DNA-BINDING DOMAINS; B VIRUS; LEUCINE ZIPPER; COILED-COIL; PARTICLES; SEQUENCE; ANTIGEN; GENE; FOS AB The capsid particle of hepadnaviruses is assembled from its dimer precursors. However, the mechanism of the protein-protein interaction is still poorly understood, A small region in the capsid protein of woodchuck hepatitis virus (WHV) contains four hydrophobic residues, including leucine 101, leucine 108, valine 115, and phenylalanine 122, that are conserved and spaced every seventh residue in the primary sequence to form a hydrophobic heptad repeat (hhr). A hydrophobic force often plays an important role in the interaction of proteins. Therefore, to investigate the role of this region in capsid assembly, we individually changed the codons specifying these four hydrophobic amino acids to codons specifying alanine or proline. In addition, we examined the in vivo infectivity of a WHV genome bearing a naturally occurring single amino acid change (histidine 104-->proline) in the hhr region. The phenotype of each altered genome was determined in both eukaryotic and prokaryotic systems by a capsid protein assay and electron microscopic examination, We show that replacement of any one of the four hydrophobic residues with alanine did not prevent capsid assembly, However, assembled capsid particles were not detected if combinations bf any two of the four residues were substituted with alanines or if the spacing of these four hydrophobic residues was changed, An individual introduction of a proline (which dramatically changes the secondary structure of proteins) into different positions of this small region also abolished capsid assembly in vitro or viral replication in vivo. These results suggested that the hhr region of the core protein of WHV was critical for capsid assembly. RP Yu, MS (reprint author), NIAID,INFECT DIS LAB,HEPATITIS VIRUSES SECT,NATL INST HLTH,7 CTR DR 0470,BETHESDA,MD 20892, USA. NR 46 TC 15 Z9 15 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0022-538X J9 J VIROL JI J. Virol. PD OCT PY 1996 VL 70 IS 10 BP 7085 EP 7091 PG 7 WC Virology SC Virology GA VG127 UT WOS:A1996VG12700068 PM 8794354 ER PT J AU Wu, WX Henderson, LE Copeland, TD Gorelick, RJ Bosche, WJ Rein, A Levin, JG AF Wu, WX Henderson, LE Copeland, TD Gorelick, RJ Bosche, WJ Rein, A Levin, JG TI Human immunodeficiency virus type 1 nucleocapsid protein reduces reverse transcriptase pausing at a secondary structure near the murine leukemia virus polypurine tract SO JOURNAL OF VIROLOGY LA English DT Article ID HAMMERHEAD RIBOZYME CATALYSIS; RNASE-H ACTIVITY; STRAND TRANSFER-REACTIONS; ACID-BINDING-PROTEINS; CYS-HIS BOX; ZINC FINGERS; VIRAL-RNA; IN-VITRO; ANNEALING ACTIVITIES; PRIMER-TEMPLATE AB In an earlier study on minus-strand DNA synthesis catalyzed by murine leukemia virus reverse transcriptase, we described a prominent pause site near the polypurine tract (J. Guo, W. Wu, Z. Y. Yuan, K. Post, R. J. Crouch, and J. G. Levin, Biochemistry 34:5018-5029, 1995). We now report that pausing at this site is due to a stem-loop structure in the RNA template, formed by interaction of a number of bases in the polypurine tract, including the six G's, and a 3' sequence which includes four C's, Addition of human immunodeficiency virus type 1 (HIV-1) nucleocapsid (NC) protein to reverse transcriptase reactions reduces pausing by similar to 8- to 10-fold and stimulates synthesis of full-length DNA. Thus, NC functions as an accessory protein during elongation of minus-strand DNA and increases the efficiency of DNA synthesis, in this case, by apparently destabilizing a region of secondary structure in the template. Since NC is associated with genomic RNA in the viral core and is likely to be part df a viral replication complex, these results suggest that NC may also promote efficient DNA synthesis during virus replication, Mutational analysis indicates that the features of HIV-1 NC which are important for reduction of pausing include the basic amino acids flanking the first zinc finger, the zinc fingers, and the cysteine and aromatic amino acids within the fingers. These findings suggest that reverse transcription might he targeted by drugs which inactivate the zinc fingers of HIV-1 NC. C1 NICHHD,GENET MOL LAB,NIH,BETHESDA,MD 20892. NCI,FREDERICK CANC RES & DEV CTR,SPECIAL PROGRAM PROT CHEM,FREDERICK,MD 21702. NCI,FREDERICK CANC RES & DEV CTR,RETROVIRAL GENET SECT,ABL BASIC RES PROGRAM,FREDERICK,MD 21702. NCI,FREDERICK CANC RES & DEV CTR,AIDS VACCINE PROGRAM,SAIC FREDERICK,FREDERICK,MD 21702. NR 81 TC 132 Z9 137 U1 0 U2 2 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0022-538X J9 J VIROL JI J. Virol. PD OCT PY 1996 VL 70 IS 10 BP 7132 EP 7142 PG 11 WC Virology SC Virology GA VG127 UT WOS:A1996VG12700074 PM 8794360 ER PT J AU Sorbara, LR Maldarelli, F Chamoun, G Schilling, B Chokekijcahi, S Staudt, L Mitsuya, H Simpson, IA Zeichner, SL AF Sorbara, LR Maldarelli, F Chamoun, G Schilling, B Chokekijcahi, S Staudt, L Mitsuya, H Simpson, IA Zeichner, SL TI Human immunodeficiency virus type 1 infection of H9 cells induces increased glucose transporter expression SO JOURNAL OF VIROLOGY LA English DT Article ID ROUS SARCOMA VIRUS; CHICKEN-EMBRYO FIBROBLASTS; SRC ONCOGENE; SUGAR TRANSPORT; T-LYMPHOCYTES; MESSENGER-RNA; GENE; TRANSFORMATION; TISSUE; RAT AB A clone obtained from a differential display screen for cellular genes with altered expression during human immunodeficiency virus (HIV) infection matched the sequence for the human GLUT3 facilitative glucose transporter, a high-velocity-high-affinity facilitative transporter commonly expressed in neurons of the central nervous system. Northern (RNA) analysis showed that GLUT3 expression increased during infection. Flow cytometry showed that GLUT3 protein expression increased specifically in the HIV-infected cells; this increase correlated with increased 2-deoxyglucose transport in the HIV-infected culture. HIV infection therefore leads to increased expression of a glucose transporter normally expressed at high Levels in other cell types and a corresponding increase in glucose transport activity. If HIV infection places increased metabolic demands on the host cell, changes in the expression of a cellular gene that plays an important role in cellular metabolism might provide a more favorable environment for viral replication. C1 NCI,PEDIAT BRANCH,NCI,BETHESDA,MD 20892. NCI,MED BRANCH,BETHESDA,MD 20892. NCI,METAB BRANCH,BETHESDA,MD 20892. NIDDKD,DIABET BRANCH,BETHESDA,MD 20892. NIAID,MOL MICROBIOL LAB,BETHESDA,MD 20892. NR 35 TC 42 Z9 43 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0022-538X J9 J VIROL JI J. Virol. PD OCT PY 1996 VL 70 IS 10 BP 7275 EP 7279 PG 5 WC Virology SC Virology GA VG127 UT WOS:A1996VG12700096 PM 8794382 ER PT J AU Heineman, TC Seidel, K Cohen, JI AF Heineman, TC Seidel, K Cohen, JI TI The varicella-zoster virus ORF66 protein induces kinase activity and is dispensable for viral replication SO JOURNAL OF VIROLOGY LA English DT Article ID HERPES-SIMPLEX VIRUS; PSEUDORABIES VIRUS; HERPESVIRUSES ENCODE; INFECTED-CELLS; SINGLE GENES; IN-VITRO; US3 GENE; TYPE-2; PHOSPHORYLATION; IDENTIFICATION AB Varicella-zoster virus (VZV) open reading frames (ORFs) 47 and 66 encode proteins that are homologous to a family of eukaryotic serine-threonine kinases, Prior studies shelved that the VZV ORF47 protein has kinase activity in vitro and is dispensable for replication in cultured cells, To examine the role of the ORF66 protein during infection, we constructed VZV recombinants that are unable to express either the ORF66 protein (ROka 66S) or both the ORF47 and ORF66 proteins (ROka 47S/66S). VZV unable to express ORF66 grew to titers similar to those of the parental VZV (ROka) in vitro; however, VZV lacking both ORF66 and ORF47 grew to titers lower than those of ROka, Nuclear extracts from ROka 66S- or ROka 47S-infected cells showed a 48-kDa phosphoprotein(s); a phosphoprotein dth a similar size was not present in nuclear extracts from ROka 47S/66S-infected cells, To determine the role of the ORF66 protein in the phosphorylation of specific VZV-encoded proteins, we immunoprecipitated known VZV phosphoproteins (ORF4, ORP62, ORP63, and ORF68 proteins) from nuclear extracts of phosphate-labeled cells infected with ROka, ROka 66S, or ROka 47S/66S. Each of the VZV phosphoproteins was phosphorylated to a similar extent in the presence or absence of either the ORF66 protein or both the ORF66 and ORF47 proteins, From these studies we conclude (i) neither ORF66 alone nor ORF66 and ORF47 in combination are essential for VZV growth in cultured cells, (ii) ORF66 either is a protein kinase or induces protein kinase activity during infection, and (iii) the VZV phosphoproteins encoded by ORF4, ORF62, ORF63, and ORF68 do not require either ORF66 alone or ORF66 and ORF47 for phosphorylation in vitro. C1 NIAID,CLIN INVEST LAB,MED VIROL SECT,BETHESDA,MD 20892. NR 34 TC 39 Z9 40 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0022-538X J9 J VIROL JI J. Virol. PD OCT PY 1996 VL 70 IS 10 BP 7312 EP 7317 PG 6 WC Virology SC Virology GA VG127 UT WOS:A1996VG12700103 PM 8794389 ER PT J AU Cho, MW Shibata, R Martin, MA AF Cho, MW Shibata, R Martin, MA TI Infection of chimpanzee peripheral blood mononuclear cells by human immunodeficiency virus type 1 requires cooperative interaction between multiple variable regions of gp120 SO JOURNAL OF VIROLOGY LA English DT Article ID ENVELOPE GLYCOPROTEIN GP120; V1/V2; GENE; LOOP AB We have recently reported the isolation and molecular cloning of a human immunodeficiency virus type 1 isolate (HIV-1(DH125)) that exhibits rapid replication kinetics and marked cytopathicity in both human and chimpanzee peripheral blood mononuclear cells (PBMC). To identify the viral determinants responsible for infectivity of chimpanzee PBMC, chimeric viruses containing the following components were constructed: (i) the entire envelope gene; (ii) gp120 sequences; (iii) gp41 sequences; and (iv) individual or various combinations of the gp120 variable regions of HIV-1(DH125) inserted into the backbone of another HIV-1 isolate (HIV-1(AD8)), which is unable to infect chimpanzee PBMC. Analyses of virus replication kinetics in human and chimpanzee PBMC revealed that gp120 contains determinants which confer infectivity for chimpanzee PBMC and that the capacity to establish such an infection requires the cooperative interaction between multiple variable regions of the HIV-1(DH125) gp120. RP Cho, MW (reprint author), NIAID,MOL MICROBIOL LAB,BLDG 4,RM 339,BETHESDA,MD 20892, USA. NR 16 TC 13 Z9 13 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0022-538X J9 J VIROL JI J. Virol. PD OCT PY 1996 VL 70 IS 10 BP 7318 EP 7321 PG 4 WC Virology SC Virology GA VG127 UT WOS:A1996VG12700104 PM 8794390 ER PT J AU Iehara, N Takeoka, H Yamada, Y Kita, T Doi, T AF Iehara, N Takeoka, H Yamada, Y Kita, T Doi, T TI Advanced glycation end products modulate transcriptional regulation in mesangial cells SO KIDNEY INTERNATIONAL LA English DT Article ID BASEMENT-MEMBRANE COLLAGEN; GLUCOSE-MODIFIED PROTEINS; ADVANCED GLYCOSYLATION; EXTRACELLULAR-MATRIX; DIABETIC NEPHROPATHY; GENE-EXPRESSION; CHLORAMPHENICOL ACETYLTRANSFERASE; METHYLATION PATTERN; MESSENGER-RNA; GROWTH-FACTOR AB Advanced glycation end products (ACEs) stimulate synthesis of extracellular matrix (ECM) in a receptor-mediated manner on mesangial cells. In the present study. we examined the transcriptional regulation of the gene for type ni collagen [(IV)collagen], which is one of the major components of mesangial sclerosis, after stimulation of AGEs on mesangial cells. The methylation pattern of the promoter:enhancer region of (IV)collagen gene was similar in AGE-treated and control cells. AGEs significantly increased the transcriptional activity of the (IV)collagen gene, as measured by transient transfection assays using the reporter gene construct containing (IV)collagen promoter:enhancer and the chloramphenicol acetyltransferase gene. AGEs also increased smooth muscle alpha-actin mRNA levels as well as its transcriptional activity. Nuclear factor binding of the promoter of (IV)collagen gene was stimulated by AGEs. Furthermore. AGEs dramatically decreased the mRNA levels of (IV)collagen promoter binding protein (MSW), a larger subunit of DNA replication complex, AP1. These results suggest that AGEs increase expression of (IV)collagen gene by modulating the levels of promoter binding proteins. These transcriptional events may play a critical role in ECM accumulation in response to AGE. C1 KYOTO UNIV,FAC MED,DIV ARTIFICIAL KIDNEYS,SAKYO KU,KYOTO 606,JAPAN. KYOTO UNIV,FAC MED,DEPT GERIATR MED,DIV CLIN BIOREGULATORY SCI,SAKYO KU,KYOTO 606,JAPAN. NIDR,DEV BIOL LAB,NIH,BETHESDA,MD 20892. NR 45 TC 24 Z9 24 U1 0 U2 3 PU BLACKWELL SCIENCE INC PI CAMBRIDGE PA 238 MAIN ST, CAMBRIDGE, MA 02142 SN 0085-2538 J9 KIDNEY INT JI Kidney Int. PD OCT PY 1996 VL 50 IS 4 BP 1166 EP 1172 DI 10.1038/ki.1996.424 PG 7 WC Urology & Nephrology SC Urology & Nephrology GA VJ314 UT WOS:A1996VJ31400011 PM 8887274 ER PT J AU Whitmore, SP Gilliam, AF Hendren, RW Lewis, SE Rao, GN Whisnant, CC AF Whitmore, SP Gilliam, AF Hendren, RW Lewis, SE Rao, GN Whisnant, CC TI Genetic monitoring of inbred rodents from controlled production colonies through biochemical markers and skin grafting procedures SO LABORATORY ANIMAL SCIENCE LA English DT Article ID MUTATIONS; MOUSE; ANTIGENS; LOCUS; MICE C1 RES TRIANGLE INST,RES TRIANGLE PK,NC 27709. NATL INST ENVIRONM HLTH SCI,RES TRIANGLE PK,NC. FU NIEHS NIH HHS [N01-ES-38044, N01-ES-65131, N01-ES-15317] NR 14 TC 2 Z9 2 U1 0 U2 0 PU AMER ASSOC LABORATORY ANIMAL SCIENCE PI CORDOVA PA 70 TIMBERCREEK DR, SUITE 5, CORDOVA, TN 38018 SN 0023-6764 J9 LAB ANIM SCI JI Lab. Anim. Sci. PD OCT PY 1996 VL 46 IS 5 BP 585 EP 588 PG 4 WC Veterinary Sciences; Zoology SC Veterinary Sciences; Zoology GA VN008 UT WOS:A1996VN00800022 PM 8905598 ER PT J AU Monahan, BP Rector, JT Liu, PP Cotelingam, JD Dahut, W AF Monahan, BP Rector, JT Liu, PP Cotelingam, JD Dahut, W TI Clinical aspects of expression of inversion 16 chromosomal fusion transcript CBFB/MYH11 in acute myelogenous leukemia subtype M1 with abnormal bone marrow eosinophilia SO LEUKEMIA LA English DT Article DE CBFB/MYH11 expression; inversion (16) chromosome; AML M1; karyotype; abnormal eosinophils; RT-PCR; prognosis ID MYELOID-LEUKEMIA; INV(16)(P13Q22) AB The pericentric inversion of chromosome 16 (inv(16)(p12q22)) is a characteristic karyotypic abnormality associated with acute myeloid leukemia, most commonly of the M4Eo subtype. It is increasingly appreciated as both a favorable prognostic factor and important guide to therapy. The breakpoints of the inversion have been cloned and a fusion transcript can be identified by RT-PCR. We expand upon our prior abstract of a case of AML subtype MI with abnormal eosinophils that expressed the inversion 16 fusion transcript CBFB/MYH11. During remission, the transcript could not be detected. The patient relapsed twice during the first year after attaining a complete remission. Morphologically, the relapsed specimens were identical to the appearance of his diagnostic specimen. However, the CBFB/MYH11 fusion transcript was not detected in the relapsed specimen. We conclude that CBFB/MYH11 fusion transcript detection, as a surrogate for the favorable prognosis and treatment planning implied by inv(16) in AML M4Eo, should not be exclusively relied upon in AML M1 in the absence of prospective study of this specific (M1) category of AML. RT-PCR analysis should be correlated with cytogenetics when available and if used alone should be interpreted cautiously in cases of AML with atypical morphological features. C1 NATL NAVAL MED CTR, DIV HEMATOL, BETHESDA, MD 20892 USA. NATL NAVAL MED CTR, DIV ONCOL, BETHESDA, MD 20007 USA. NIH, LAB GENE TRANSFER, NATL CTR HUMAN GENOME RES, BETHESDA, MD USA. GEORGETOWN UNIV, MED CTR, LOMBARDI CANC CTR, WASHINGTON, DC USA. RI Liu, Paul/A-7976-2012 OI Liu, Paul/0000-0002-6779-025X NR 9 TC 4 Z9 4 U1 0 U2 0 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE, HAMPSHIRE, ENGLAND RG21 6XS SN 0887-6924 J9 LEUKEMIA JI Leukemia PD OCT PY 1996 VL 10 IS 10 BP 1653 EP 1654 PG 2 WC Oncology; Hematology SC Oncology; Hematology GA VL641 UT WOS:A1996VL64100012 PM 8847901 ER PT J AU Bergasa, NV Vergalla, J Swain, MG Jones, EA AF Bergasa, NV Vergalla, J Swain, MG Jones, EA TI Hepatic concentrations of proenkephalin-derived opioids are increased in a rat model of cholestasis SO LIVER LA English DT Article DE bile duct resection; cholestasis; endogenous opioids; hepatic concentration; preproenkephalin ID PRIMARY BILIARY-CIRRHOSIS; METHIONINE-ENKEPHALIN; CONTROLLED TRIAL; NALOXONE INFUSIONS; NALMEFENE THERAPY; DOUBLE-BLIND; PEPTIDES; PRURITUS; PLASMA; LIVER AB The liver of adult rats with cholestasis secondary to bile duct resection has been shown to express the proenkephalin gene and, by immunohistochemical stains, to contain met-enkephalin. To further study hepatic opioids in cholestasis, concentrations of proenkephalin-derived endogenous opioids were measured in a rat model of cholestasis by the use of radioimmunoassays. The specificity of the immunoreactivity detected by the assays was confirmed by high performance liquid chromatography (HPLC). In adult male rats with cholestasis due to BDR, the concentrations of three proenkephalin-derived opioid peptides were increased. Specifically, the mean hepatic concentrations of met-enkephalin, Met-Enk-Arg(6)-Phe(7) and leu-enkephalin were 2.5 (p<0.005), 2.1 (p<0.005) and 2.5 (p<0.01) fold higher than the corresponding mean for controls. These findings provide further independent evidence that opioid peptides accumulate in the liver in a model of cholestasis and are consistent with de novo synthesis of opioid peptides occurring in the cholestatic liver. This phenomenon may have relevance to the altered function of the opioid system in cholestasis and to the role of the liver as a neuroendocrine organ. C1 NATL INST HLTH,LIVER DIS SECT,BETHESDA,MD. NR 34 TC 39 Z9 39 U1 0 U2 0 PU MUNKSGAARD INT PUBL LTD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0106-9543 J9 LIVER JI Liver PD OCT PY 1996 VL 16 IS 5 BP 298 EP 302 PG 5 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA VU743 UT WOS:A1996VU74300002 PM 8938629 ER PT J AU Lockshin, MD AF Lockshin, MD TI Pathogenesis of the antiphospholipid antibody syndrome SO LUPUS LA English DT Article; Proceedings Paper CT 7th International Symposium on Antiphospholipid Antibodies CY OCT 09-13, 1996 CL NEW ORLEANS, LA DE antiphospholipid antibody; beta(2)-glycoprotein-I; endothelium; platelets; placenta ID PLACENTAL ANTICOAGULANT PROTEIN; SYSTEMIC LUPUS-ERYTHEMATOSUS; BETA-2 GLYCOPROTEIN-I; ANTICARDIOLIPIN ANTIBODIES; ENDOTHELIAL-CELLS; FETAL LOSS; PLATELET ACTIVATION; BINDING; BETA(2)-GLYCOPROTEIN-I; ASPIRIN AB Endothelial and/or platelet activation likely initiates thrombus formation. Whether antiphospholipid antibody (aPL) is an activator, a toxic response, or a protective response is not clear, nor is it certain whether aPL is germ-line encoded or antigen-driven. The pregnancy model is particularly informative. Alternative hypotheses about thrombogenicity which relegate aPL to the role of bystander have not yet been excluded. RP Lockshin, MD (reprint author), NIH,WARREN GRANT MAGNUSON CLIN CTR,BLDG 10,ROOM 2C-146,10 CTR DR,MSC 1504,BETHESDA,MD 20892, USA. NR 43 TC 26 Z9 26 U1 0 U2 0 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE, HAMPSHIRE, ENGLAND RG21 6XS SN 0961-2033 J9 LUPUS JI Lupus PD OCT PY 1996 VL 5 IS 5 BP 404 EP 408 PG 5 WC Rheumatology SC Rheumatology GA VQ415 UT WOS:A1996VQ41500014 PM 8902770 ER PT J AU Yang, YH Glover, GH vanGelderen, P Mattay, VS Santha, AKS Sexton, RH Ramsey, NF Moonen, CTW Weinberger, DR Frank, JA Duyn, JH AF Yang, YH Glover, GH vanGelderen, P Mattay, VS Santha, AKS Sexton, RH Ramsey, NF Moonen, CTW Weinberger, DR Frank, JA Duyn, JH TI Fast 3D functional magnetic resonance imaging at 1.5 T with spiral acquisition SO MAGNETIC RESONANCE IN MEDICINE LA English DT Article DE MRI; spiral scanning; fMRI; motor cortex ID HUMAN BRAIN; MOTOR CORTEX; SENSORY STIMULATION; GRADIENT ECHOES; ACTIVATION; SIGNAL; CONTRAST; MRI; NMR; OXYGENATION AB A new method to perform rapid 3D fMRI in human brain is introduced and evaluated in normal subjects, on a standard clinical scanner at 1.5 Tesla, The method combines a highly stable gradient echo technique with a spiral scan method, to detect brain activation related changes in blood oxygenation with high sensitivity. A motor activation paradigm with a duration of less than 5 min, performed on 10 subjects, consistently showed significant changes in signal intensity in the area of the motor cortex. In all subjects, these changes survived high statistical thresholds. C1 NIH,OIR,LAB DIAGNOST RADIOL RES,BETHESDA,MD 20892. STANFORD UNIV,SCH MED,LUCAS MRI CTR,DEPT DIAGNOST RADIOL,STANFORD,CA 94305. NIH,IN VITRO NMR RES CTR,BEIP,NCRR,BETHESDA,MD. NIMH,CLIN BRAIN DISORDERS BRANCH,NIH,BETHESDA,MD 20892. RI Duyn, Jozef/F-2483-2010; Moonen, Chrit/K-4434-2016 OI Moonen, Chrit/0000-0001-5593-3121 NR 45 TC 52 Z9 53 U1 0 U2 0 PU WILLIAMS & WILKINS PI BALTIMORE PA 351 WEST CAMDEN ST, BALTIMORE, MD 21201-2436 SN 0740-3194 J9 MAGNET RESON MED JI Magn.Reson.Med. PD OCT PY 1996 VL 36 IS 4 BP 620 EP 626 DI 10.1002/mrm.1910360418 PG 7 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA VL633 UT WOS:A1996VL63300017 PM 8892216 ER PT J AU Andersson, L Archibald, A Ashburner, M Audun, S Barendse, W Bitgood, J Bottema, C Broad, T Brown, S Burt, D Charlier, C Copeland, N Davis, S Davisson, M Edwards, J Eggen, A Elgar, G Eppig, JT Franklin, I Grewe, P Gill, T Graves, JAM Hawken, R Hetzel, J Hilyard, A Jacob, H Jaswinska, L Jenkins, N Kunz, H Levan, G Lie, O Lyons, L Maccarone, P Mellersh, C Montgomery, G Moore, S Moran, C Morizot, D Neff, M Nicholas, F OBrien, S Parsons, Y Peters, J Postlethwait, J Raymond, M Rothschild, M Schook, L Sugimoto, Y Szpirer, C Tate, M Taylor, J VandeBerg, J Wakefield, M Wienberg, J Womack, J AF Andersson, L Archibald, A Ashburner, M Audun, S Barendse, W Bitgood, J Bottema, C Broad, T Brown, S Burt, D Charlier, C Copeland, N Davis, S Davisson, M Edwards, J Eggen, A Elgar, G Eppig, JT Franklin, I Grewe, P Gill, T Graves, JAM Hawken, R Hetzel, J Hilyard, A Jacob, H Jaswinska, L Jenkins, N Kunz, H Levan, G Lie, O Lyons, L Maccarone, P Mellersh, C Montgomery, G Moore, S Moran, C Morizot, D Neff, M Nicholas, F OBrien, S Parsons, Y Peters, J Postlethwait, J Raymond, M Rothschild, M Schook, L Sugimoto, Y Szpirer, C Tate, M Taylor, J VandeBerg, J Wakefield, M Wienberg, J Womack, J TI Comparative genome organization of vertebrates SO MAMMALIAN GENOME LA English DT Article ID GENETIC-LINKAGE MAP; IN-SITU HYBRIDIZATION; WALLABY MACROPUS-EUGENII; QUANTITATIVE TRAIT LOCI; B RECEPTOR GENE; PIG SUS SCROFA; X-CHROMOSOME; REGIONAL LOCALIZATION; RATTUS-NORVEGICUS; GALLUS-DOMESTICUS C1 UNIV CAMBRIDGE,DEPT GENET,CAMBRIDGE CB2 3EH,ENGLAND. NORWEGIAN COLL VET MED,DEPT MORPHOL GENET & AQUAT BIOL,N-0033 OSLO,NORWAY. UNIV QUEENSLAND,DIV TROP ANIM PROD,MOL ANIM GENET CTR,ST LUCIA,QLD 4072,AUSTRALIA. UNIV WISCONSIN,DEPT POULTRY SCI,MADISON,WI 53706. UNIV ADELAIDE,WAITE AGR RES INST,DEPT ANIM SCI,GLEN OSMOND,SA 5064,AUSTRALIA. ST MARYS HOSP,SCH MED,DEPT BIOCHEM & MOL GENET,LONDON W2 1PG,ENGLAND. AGRES,INVERNAY AGR CTR,MOSGIEL,NEW ZEALAND. FAC MED,DEPT GENET,B-4000 LIEGE,BELGIUM. NCI,FREDERICK CANC RES & DEV CTR,ABL BASIC RES PROGRAM,FREDERICK,MD 21702. TEXAS A&M UNIV,DEPT ANIM SCI,COLLEGE STN,TX 77843. JACKSON LAB,BAR HARBOR,ME 04609. ABS GLOBAL INC,DE FOREST,WI 53532. ADDENBROOKES HOSP,DEPT MED,CAMBRIDGE CB2 2QQ,ENGLAND. JACKSON LAB,BAR HARBOR,ME 04609. CSIRO,DIV ANIM PROD,BLACKTOWN,NSW 2148,AUSTRALIA. CSIRO,HOBART,TAS 7001,AUSTRALIA. UNIV PITTSBURGH,DEPT PATHOL,PITTSBURGH,PA 15261. LA TROBE UNIV,SCH GENET & HUMAN VARIAT,BUNDOORA,VIC 3083,AUSTRALIA. UNIV MELBOURNE,SCH VET SCI,CTR ANIM BIOTECHNOL,MELBOURNE,VIC 3052,AUSTRALIA. UNIV QUEENSLAND,MOL ANIM GENET CTR,DIV TROP ANIM PROD,CSIRO,ST LUCIA,QLD 4072,AUSTRALIA. JACKSON LAB,BAR HARBOR,ME 04609. MASSACHUSETTS GEN HOSP EAST,CARDIOVASC RES CTR,CHARLESTOWN,MA. UNIV QUEENSLAND,ROYAL BRISBANE HOSP,BANCROFT CTR,QUEENSLAND INST MED RES,HERSTON,QLD,AUSTRALIA. NCI,FREDERICK CANC RES & DEV CTR,ABL BASIC RES PROGRAM,FREDERICK,MD 21702. UNIV PITTSBURGH,SCH MED,DEPT PATHOL,PITTSBURGH,PA 15261. DEPT GENET,S-41390 GOTHENBURG,SWEDEN. NORWEGIAN COLL VET MED,DEPT MORPHOL GENET & AQUAT BIOL,DIV GENET,N-0033 OSLO,NORWAY. NCI,FREDERICK CANC RES & DEV CTR,LVC,FREDERICK,MD 21702. LA TROBE UNIV,SCH BIOL SCI,DEPT GENET & HUMAN VARIAT,BUNDOORA,VIC 3083,AUSTRALIA. FRED HUTCHINSON CANC RES CTR,DIV CLIN M318,SEATTLE,WA 98104. UNIV OTAGO,DEPT BIOCHEM,AGRES,MOL BIOL UNIT,DUNEDIN,NEW ZEALAND. UNIV QUEENSLAND,CSIRO,DIV TROP ANIM PROD,MOL ANIM GENET CTR,ST LUCIA,QLD 4072,AUSTRALIA. UNIV SYDNEY,DEPT ANIM SCI,SYDNEY,NSW 2006,AUSTRALIA. UNIV TEXAS,MD ANDERSON CANC CTR,DIV RES,SMITHVILLE,TX 78957. UNIV CALIF BERKELEY,DEPT MOL & CELLULAR BIOL,BERKELEY,CA 94720. NCI,VIRAL CARCINOGENESIS LAB,FREDERICK,MD 21702. MACQUARIE UNIV,SCH BIOL SCI,N RYDE,NSW 2109,AUSTRALIA. MRC,MAMMALIAN GENET UNIT,DIDCOT,OXON,ENGLAND. UNIV OREGON,INST NEUROSCI,EUGENE,OR 97403. NCI,FREDERICK CANC RES & DEV CTR,VIRAL CARCINOGENESIS LAB,GENET SECT,FREDERICK,MD 21702. IOWA STATE UNIV SCI & TECHNOL,DEPT ANIM SCI,AMES,IA 50011. UNIV MINNESOTA,DEPT VET PATHOBIOL,ST PAUL,MN 55108. SHIRAKAWA INST ANIM GENET,FUKUSHIMA 961,JAPAN. FREE UNIV BRUSSELS,DEPT BIOL MOL,B-1640 RHODE ST GENESE,BELGIUM. TEXAS A&M UNIV,DEPT ANIM SCI,COLLEGE STN,TX 77843. SW FDN BIOMED RES,SAN ANTONIO,TX 78245. LA TROBE UNIV,SCH GENET & HUMAN VARIAT,BUNDOORA,VIC 3083,AUSTRALIA. UNIV CAMBRIDGE,DEPT PATHOL,CAMBRIDGE CB2 1QP,ENGLAND. TEXAS A&M UNIV,DEPT VET PATHOBIOL,COLLEGE STN,TX 77843. ROSLIN INST,DEPT MOL BIOL,ROSLIN EH25 9PS,MIDLOTHIAN,SCOTLAND. RP Andersson, L (reprint author), SWEDISH UNIV AGR SCI,DEPT ANIM BREEDING & GENET,UPPSALA,SWEDEN. RI Wakefield, Matthew/A-7795-2008; Graves, Jennifer/A-1387-2008; Barendse, William/D-8608-2011; Grewe, Peter/P-1515-2015; OI Wakefield, Matthew/0000-0001-6624-4698; Barendse, William/0000-0002-5464-0658; Grewe, Peter/0000-0001-7111-4150; Charlier, Carole/0000-0002-9694-094X; Schook, Lawrence/0000-0002-6580-8364 NR 89 TC 113 Z9 116 U1 0 U2 2 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 SN 0938-8990 J9 MAMM GENOME JI Mamm. Genome PD OCT PY 1996 VL 7 IS 10 BP 717 EP 734 DI 10.1007/s003359900222 PG 18 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA VL722 UT WOS:A1996VL72200002 PM 8854859 ER PT J AU Lock, LF Wickramasinghe, D Ernst, MK Gilbert, DJ Copeland, NG Jenkins, NA Donovan, PJ AF Lock, LF Wickramasinghe, D Ernst, MK Gilbert, DJ Copeland, NG Jenkins, NA Donovan, PJ TI The Cdc25 genes map to mouse chromosomes 2, 9, and 18 SO MAMMALIAN GENOME LA English DT Article ID LINKAGE MAP; PHOSPHATASES; CLONING C1 NCI,MAMMALIAN GENET LAB,ABL BASIC RES PROGRAM,FREDERICK CANC RES & DEV CTR,FREDERICK,MD 21702. NR 14 TC 5 Z9 5 U1 0 U2 0 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 SN 0938-8990 J9 MAMM GENOME JI Mamm. Genome PD OCT PY 1996 VL 7 IS 10 BP 771 EP 772 DI 10.1007/s003359900230 PG 2 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA VL722 UT WOS:A1996VL72200010 PM 8854867 ER PT J AU Kozak, CA Sangameswaran, L AF Kozak, CA Sangameswaran, L TI Genetic mapping of the peripheral sodium channel genes, Scn9a and Scn10a, in the mouse SO MAMMALIAN GENOME LA English DT Article C1 ROCHE BIOSCI,INST PHARMACOL,NEUROBIOL UNIT,PALO ALTO,CA 94304. RP Kozak, CA (reprint author), NIAID,MOL MICROBIOL LAB,NIH,BLDG 4,ROOM 329,BETHESDA,MD 20892, USA. NR 13 TC 15 Z9 15 U1 0 U2 1 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 SN 0938-8990 J9 MAMM GENOME JI Mamm. Genome PD OCT PY 1996 VL 7 IS 10 BP 787 EP 788 DI 10.1007/s003359900235 PG 2 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA VL722 UT WOS:A1996VL72200015 PM 8854872 ER PT J AU Nagi, DK Tracy, R Pratley, R AF Nagi, DK Tracy, R Pratley, R TI Relationship of hepatic and peripheral insulin resistance with plasminogen activator inhibitor-1 in Pima Indians SO METABOLISM-CLINICAL AND EXPERIMENTAL LA English DT Article ID DEPENDENT DIABETES-MELLITUS; HUMAN ENDOTHELIAL-CELLS; CARDIOVASCULAR-DISEASE; GLUCOSE; PAI-1; CAUCASIANS; EXPRESSION; SECRETION; ANTIGEN; OBESITY AB Plasminogen activator inhibitor-1 (PAI-1) is related to insulin resistance and several components of the insulin resistance syndrome, and PAI-1 levels are elevated in subjects with non-insulin-dependent diabetes mellitus. Many Pima Indians are obese, insulin-resistant, and hyperinsulinemic, and they have high rates of diabetes but a low risk of ischemic heart disease. In contrast to whites and Asians, PAI-1 activity is similar between nondiabetic and diabetic Pima Indians. We therefore examined the association of PAI-1 with hepatic and peripheral insulin action measured using the hyperinsulinemic-euglycemic clamp. To investigate if insulin per se has any effect on PAI-1 in vivo, we also assessed the effects of endogenous (during a 75-g oral glucose load) and exogenous (during hyperinsulinemic clamp) insulin on PAI-1 antigen. Twenty-one (14 men and seven women; mean age, 26.3 +/- 4.8 years) Pima Indians underwent a 75-g oral glucose tolerance test (OGTT) and a sequential hyperinsulinemic-euglycemic clamp. Peripheral insulin action was measured as absolute glucose uptake (M value) and normalized to estimated metabolic body size (EMBS). Hepatic insulin action was measured as percent suppression of basal hepatic glucose output during hyperinsulinemia. PAI-1 antigen was determined using a two-site enzyme-linked immunosorbent assay that detects only free PAI-1. PAI-1 antigen concentrations were significantly related to body mass index ([BMI] r(s) = .54, P = .012), waist (r(s) = .52, P = .016) and thigh (r(s) = .63, P = .002) circumference, and fasting plasma insulin concentration (r(s) = .59, P = .004). PAI-1 antigen concentrations were not significantly associated with peripheral glucose uptake (M value) during either low-dose (r(s) = -.01, P = NS) or high-dose (r(s) = -.11, P = NS) insulin infusion. PAI-1 antigen was negatively correlated with basal hepatic glucose output (P = -.57, P = .013) and percent suppression of hepatic glucose output during hyperinsulinemia (r(s) = -.69, P = .005). However, this relationship was largely due to the confounding effects of BMI, waist and thigh girth, fasting insulin, and 2-hour postload glucose concentrations, and was not significant when controlled for these variables (partial r(s) = -.30, P = NS). There was no significant relationship of PAI-1 antigen concentration with glucose storage or glucose oxidation. Despite a threefold increase in plasma insulin concentrations during the OGTT, there were no significant changes in PAI-1 antigen concentrations (median, 57, 61, 55, and 44 ng/mL at 0, 60, 120, and 180 minutes, respectively; P = NS by ANOVA). During the hyperinsulinemic clamp, mean plasma insulin concentrations at the end of low-dose (240 pmol/m(2)/min) and high-dose (2,400 pmol/m(2)/min) infusions were 1,005 and 14,230 pmol/L, respectively. However, PAI-1 antigen concentrations at the end of low-dose and high-dose insulin infusions were similar to those at baseline (median, 63, 43, and 58 ng/mL, respectively; P = NS by ANOVA). PAI-1 antigen in Pima Indians is related to several components of the insulin resistance syndrome. However, direct measurement of insulin resistance indicates that hepatic but not peripheral insulin resistance is related to PAI-1 antigen. Neither endogenous nor exogenous hyperinsulinemia for short periods had any significant effect on PAI-1 antigen concentrations. Short-term hyperinsulinemia is unlikely to be an important regulator of PAI-1 in Pima Indians. The relationship of PAI-1 antigen to hepatic insulin resistance is largely dependent on the relationship of PAI-1 to indices of obesity and fasting insulin concentrations. Copyright (C) 1996 by W.B. Saunders Company C1 NIDDKD,NIH,PHOENIX,AZ 85016. THROMBOSIS RES CTR,BURLINGTON,VT. NR 41 TC 21 Z9 21 U1 0 U2 1 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0026-0495 J9 METABOLISM JI Metab.-Clin. Exp. PD OCT PY 1996 VL 45 IS 10 BP 1243 EP 1247 DI 10.1016/S0026-0495(96)90242-5 PG 5 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VK866 UT WOS:A1996VK86600009 PM 8843179 ER PT J AU Masuelli, L Cutler, ML AF Masuelli, L Cutler, ML TI Increased expression of the ras suppressor Rsu-1 enhances Erk-2 activation and inhibits jun kinase activation SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID ACTIN STRESS FIBERS; SIGNALING PATHWAY; PROTEIN-KINASE; SACCHAROMYCES-CEREVISIAE; ADENYLYL CYCLASE; CDC42 GTPASES; GENE-PRODUCT; 3T3 CELLS; H-RAS; TRANSFORMATION AB Studies were undertaken to determine the effect of the Pas suppressor Rsu-1 on Ras signal transduction pathways in two different cell backgrounds. An expression vector containing the mouse rsu-1 cDNA under the control of a mouse mammary tumor virus promoter mas introduced into NIH 3T3 cells and the pheochromocytoma cell line PC12. Cell lines developed in the NIH 3T3 background expressed p33(rsu-1) at approximately twice the normal endogenous level. However, PC12 cell clones which expressed p33(rsu-1) at an increased level in a regulatable fashion in response to dexamethasone were isolated. Analysis of proteins involved in regulation of Pas and responsive to Pas signal transduction revealed similar changes in the two cell backgrounds in the presence of elevated p33(rsu-1). There was an increase in the level of SOS, the guanine nucleotide exchange factor, and an increase in the percentage of GTP-bound Ras. In addition, there was an increase in the amount of p120 Pas-specific GTPase-activating protein (GAP) and GAP-associated p190. However, a decrease in Ras GTPase-activating activity spas detected in lysates of the Rsu-1 transfectants, and immunoprecipitated p120 GAP from the Rsu-1 transfectants showed less Ras GTPase-activating activity than GAP from control cells. Activation of Erk-2 kinase by growth factor and tetradecanyol phorbol acetate was greater in the Rsu-1 transfectants than in control cells. However, c-Jun amino-terminal kinase activity (Jun kinase) was not activatable bg epidermal growth factor in Rsu-1 PC12 cell transfectants, in contrast to the PC12 vector control cell line, Transient expression of p33(rsu-1) in Cos1 cells following cotransfection with either hemagglutinin-tagged Jun kinase or hemagglutinin-tagged Erk-2 revealed that Rsu-1 expression inhibited constitutive Jun kinase activity while enhancing Erk-2 activity. Detection of in vitro binding of Rsu-1 to Raf-1 suggested that in Rsu-1 transfectants, increased activation of the Raf-1 pathway occurred at the expense of activation of signal transduction leading to Jun kinase. These results indicate that inhibition of Jun kinase activation was sufficient to inhibit Ras transformation even in the presence of activated Erk-2. C1 UNIFORMED SERV UNIV HLTH SCI,DEPT PATHOL,BETHESDA,MD 20814. NCI,TUMOR IMMUNOL & BIOL LAB,BETHESDA,MD 20892. NR 45 TC 34 Z9 37 U1 0 U2 2 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0270-7306 J9 MOL CELL BIOL JI Mol. Cell. Biol. PD OCT PY 1996 VL 16 IS 10 BP 5466 EP 5476 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA VH852 UT WOS:A1996VH85200026 PM 8816460 ER PT J AU Jackson, BM Drysdale, CM Natarajan, K Hinnebusch, AG AF Jackson, BM Drysdale, CM Natarajan, K Hinnebusch, AG TI Identification of seven hydrophobic clusters in GCN4 making redundant contributions to transcriptional activation SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID RNA-POLYMERASE-II; TATA-BINDING PROTEIN; THYROID-HORMONE RECEPTOR; AMINO-ACID CONTROL; IN-VIVO; BASAL TRANSCRIPTION; FACTOR-TFIIB; SACCHAROMYCES-CEREVISIAE; TRANSACTIVATION DOMAIN; GENETIC-EVIDENCE AB GCN4 is a transcriptional activator in the bZIP family that regulates amino acid biosynthetic genes in the yeast Saccharomyces cerevisiae. The N-terminal 100 amino acids of GCN4 contains a potent activation function that confers high-level transcription in the absence of the centrally located acidic activation domain (CAAD) delineated in previous studies. To identify specific amino acids important for activation by the N-terminal domain, we mutagenized a GCN4 allele lacking the CAAD and screened alleles in vivo for reduced expression of the HIS3 gene. We found four pairs of closely spaced phenylalanines and a leucine residue: distributed throughout the N-terminal 100 residues of GCN4 that are required for high-level activation in the absence of the CAAD. Trp, Leu, and Tyr were highly functional substitutions for the Phe residue at position 45. Combined with our previous findings, these results indicate that GCN4 contains seven clusters of aromatic or bulky hydrophobic residues which make important contributions to transcriptional activation at HIS3. None of the seven hydrophobic clusters is essential for activation by full-length GCN4, and the critical residues in two or three clusters must be mutated simultaneously to observe a substantial reduction in GCN4 function, Numerous combinations of four or five intact clusters conferred high-level transcription of HIS3. We propose that many of the hydrophobic clusters in GCN4 act independently of one another to provide redundant means of stimulating transcription and that the functional contributions of these different segments are cumulative at the HIS3 promoter. On the basis of the primacy of bulky hydrophobic residues throughout the activation domain, we suggest that GCN4 contains multiple sites that mediate hydrophobic contacts with one or more components of the transcription initiation machinery. C1 NICHHD, LAB EUKARYOT GENE REGULAT, BETHESDA, MD 20892 USA. NR 73 TC 61 Z9 62 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0270-7306 J9 MOL CELL BIOL JI Mol. Cell. Biol. PD OCT PY 1996 VL 16 IS 10 BP 5557 EP 5571 PG 15 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA VH852 UT WOS:A1996VH85200034 PM 8816468 ER PT J AU Levin, HL AF Levin, HL TI An unusual mechanism of self-primed reverse transcription requires the RNase H domain of reverse transcriptase to cleave an RNA duplex SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID RETROVIRUS-LIKE PARTICLES; VIRUS-LIKE PARTICLES; CELL-FREE SYNTHESIS; RIBONUCLEASE-H; TY1 TRANSPOSITION; ESCHERICHIA-COLI; FISSION YEAST; LINKED MSDNA; SCHIZOSACCHAROMYCES-POMBE; ELEMENT TRANSPOSITION AB The reverse transcription of retroviruses and long terminal repeat-containing retrotransposons requires that tRNA species serve as primers. We recently reported that the long terminal repeat-containing retrotransposon Tf1 is a unique exception in that reverse transcription is independent of tRNA and is instead initiated by a self-priming mechanism. The first 11 bases of the Tf1 transcript fold back and anneal to the primer binding site in a process that results in the priming of minus-strand strong-stop DNA. Data presented here demonstrate that a cleavage occurs between the 11th and 12th bases of the transcript, resulting in the generation of the primer. Mutagenesis experiments presented here indicate that the RNase H domain of the Tf1 reverse transcriptase is required for the cleavage reaction, suggesting that this RNase H may have the novel ability to cleave double-stranded RNA at the end of a duplexed region. RP NICHHD, LAB EUKARYOT GENE REGULAT, BETHESDA, MD 20892 USA. NR 45 TC 48 Z9 49 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0270-7306 EI 1098-5549 J9 MOL CELL BIOL JI Mol. Cell. Biol. PD OCT PY 1996 VL 16 IS 10 BP 5645 EP 5654 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA VH852 UT WOS:A1996VH85200043 PM 8816477 ER PT J AU Moriggl, R GouilleuxGruart, V Jahne, R Berchtold, S Gartmann, C Liu, XW Hennighausen, L Sotiropoulos, A Groner, B Gouilleux, F AF Moriggl, R GouilleuxGruart, V Jahne, R Berchtold, S Gartmann, C Liu, XW Hennighausen, L Sotiropoulos, A Groner, B Gouilleux, F TI Deletion of the carboxyl-terminal transactivation domain of MGF-Stat5 results in sustained DNA binding and a dominant negative phenotype SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID CASEIN GENE PROMOTER; MAMMARY-GLAND FACTOR; RECEPTOR-BETA-CHAIN; COLONY-STIMULATING FACTOR; TRANSCRIPTION FACTOR; NUCLEAR FACTOR; TYROSINE PHOSPHORYLATION; SIGNALING MOLECULES; CYTOKINE RECEPTORS; STAT5-LIKE FACTOR AB The Stat (signal transducer and activator of transcription) factors transmit cytokine, growth factor, and hormone responses, Seven members of the Stat gene family are known, MGF-Stat5a has been discovered as a mediator of the prolactin response in mammary epithelial cells. Two closely related variants of Stat5, Stat5a and Stat5b, are encoded by distinct genes. We examined the functional properties of the carboxyl termini of these molecules, Wild-type Stat5a (794 amino acids) and the carboxyl-terminal deletion mutant Stat5a Delta 772 supported prolactin-induced transcription of a beta-casein promoter-reporter construct in COS7 cells; Stat5a Delta 750 did not, Upon prolactin activation, tyrosine phosphorylation and the specificity of DNA binding were indistinguishable among the three Stat5a variants, Tyrosine dephosphorylation and the downregulation of the DNA-binding activity were delayed in the Stat5a Delta 750 mutant, The carboxyl-terminal transactivation domain of Stat5a, amino acids 722 to 794, can be conferred to the DNA-binding domain of the yeast transcription factor GAL4. Coexpression of Stat5a or Stat5b and of the carboxyl-terminal deletion mutants resulted in the suppression of transcriptional induction in COS or Ba/F3 cells. We propose that Stat5a Delta 750 and Stat5b Delta 754 are lacking functional transactivation domains and exert their dominant negative effects by blocking the DNA-binding site in Stat5-responsive gene promoters. C1 UNIV FREIBURG, TUMOR BIOL CTR, INST EXPT CANC RES, D-79106 FREIBURG, GERMANY. UNIV FREIBURG, DEPT BIOL, D-79106 FREIBURG, GERMANY. MAX PLANCK INST IMMUNBIOL, D-79108 FREIBURG, GERMANY. CTR HOSP AMIENS, IMMUNOL LAB, F-80000 AMIENS, FRANCE. HOP NECKER ENFANTS MALAD, FAC MED, INSERM U344, F-75730 PARIS 15, FRANCE. NIDDK, BIOCHEM & METAB LAB, BETHESDA, MD 20892 USA. NR 52 TC 218 Z9 219 U1 2 U2 3 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0270-7306 EI 1098-5549 J9 MOL CELL BIOL JI Mol. Cell. Biol. PD OCT PY 1996 VL 16 IS 10 BP 5691 EP 5700 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA VH852 UT WOS:A1996VH85200048 PM 8816482 ER PT J AU Schulte, TW Blagosklonny, MV Romanova, L Mushinski, JF Monia, BP Johnston, JF Nguyen, P Trepel, J Neckers, LM AF Schulte, TW Blagosklonny, MV Romanova, L Mushinski, JF Monia, BP Johnston, JF Nguyen, P Trepel, J Neckers, LM TI Destabilization of Raf-1 by geldanamycin leads to disruption of the Raf-1-MEK-mitogen-activated protein kinase signalling pathway SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID EPIDERMAL GROWTH-FACTOR; HEAT-SHOCK PROTEINS; MAP KINASE; GENE-EXPRESSION; ANTISENSE OLIGONUCLEOTIDES; TRANSCRIPTIONAL ACTIVITY; PHOSPHORYLATION SITES; COMPLEX-FORMATION; PLASMA-MEMBRANE; PC12 CELLS AB The serine/threonine kinase Raf-1 functions downstream of Ras in a signal transduction cascade which transmits mitogenic stimuli from the plasma membrane to the nucleus. Raf-1 integrates signals coming from extracellular factors and, in turn, activates its substrate, MEK kinase. MEK activates mitogen-activated protein kinase (MAPK), which phosphorylates other kinases as well as transcription factors. Raf-1 exists in a complex with HSP90 and other proteins. The benzoquinone ansamycin geldanamycin (GA) binds to HSP90 and disrupts the Raf-1-HSP90 multimolecular complex, leading to destabilization of Raf-1. In this study, we examined whether Raf-1 destabilization is sufficient to block the Raf-1-MEK-MAPK signalling pathway and whether GA specifically inactivates the Raf-1 component of this pathway. Using the model system of NIH 3T3 cells stimulated with phorbol 12-myristate 13-acetate (PMA), we show that GA does not affect the ability of protein kinase Co to be activated by phorbol esters, but it does block activation of MEK and MAPK. Further, GA does not decrease the activity of constitutively active MEK in transiently transfected cells. Finally, disruption of the Raf-1-MEK-MAPK signalling pathway by GA prevents both the PMA-induced proliferative response and PMA-induced activation of a MAPK-sensitive nuclear transcription factor. Thus, we demonstrate that interaction between HSP90 and Raf-1 is a sine qua non for Raf stability and function as a signal transducer and that the effects observed cannot be attributed to a general impairment of protein kinase function. C1 NCI,GENET LAB,BETHESDA,MD 20892. NCI,MED ONCOL BRANCH,BETHESDA,MD 20892. ISIS PHARMACEUT,DEPT MOL PHARMACOL,CARLSBAD,CA 92008. RP Schulte, TW (reprint author), NCI,CLIN PHARMACOL BRANCH,BLDG 10,ROOM 13N240,10 CTR DR,MSC 1928,BETHESDA,MD 20892, USA. NR 56 TC 217 Z9 221 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 SN 0270-7306 J9 MOL CELL BIOL JI Mol. Cell. Biol. PD OCT PY 1996 VL 16 IS 10 BP 5839 EP 5845 PG 7 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA VH852 UT WOS:A1996VH85200064 PM 8816498 ER PT J AU Moore, KD DillonCarter, O Conejero, C Poltorak, M Chedid, M Tornatore, C Freed, WJ AF Moore, KD DillonCarter, O Conejero, C Poltorak, M Chedid, M Tornatore, C Freed, WJ TI In vitro properties of a newly established medulloblastoma cell line, MCD-1 SO MOLECULAR AND CHEMICAL NEUROPATHOLOGY LA English DT Article DE medulloblastoma; cyclic AMP; differentiation; glutamate; nerve growth factor; p53; TGF beta; CNS tumors ID NERVE GROWTH-FACTOR; FIBRILLARY ACIDIC PROTEIN; PRIMITIVE NEUROECTODERMAL TUMORS; NEURON-SPECIFIC ENOLASE; FACTOR-BETA; CYCLIC-AMP; ASTROCYTIC DIFFERENTIATION; NEUROFILAMENT PROTEIN; CULTURED ASTROCYTES; ASTROGLIAL CELLS AB Medulloblastomas are poorly differentiated brain tumors believed to arise from primitive pleuripotential stem cells, and tend to express mixed neuronal and glial properties. In the present study, we examined immunohistochemical and neurotransmitter phenotypic properties in a newly established medulloblastoma cell line, MCD-1. MCD-1 cells were immortal, not contact-inhibited, but did not grow in soft agar. Immunohistochemical studies showed positive staining for neurofilament protein (NF), neuron-specific enolase (NSE), synaptophysin, MAP 2, tau, NCAM 180, vimentin, and S-100 protein. The cells expressed specific uptake of glutamate, serotonin, and choline, but not GABA or dopamine. A significant increase in process extension was seen in response to agents that enhance intracellular cyclic AMP, especially 3-isobutyl-1-methylxanthine (IBMX). Process formation induced by IBMX was associated with a decrease in cell proliferation as evidenced by a reduction in numbers of cells incorporating 5-bromo-2-deoxyuridine (BrdU). No increase in process extension was observed following exposure to NGF or retinoic acid. MCD-1 cells were shown to produce transforming growth factor beta (TGF beta), and were immunopositive for mutant p53. Transfection assays with the PG13-Luc reporter plasmid, which contains a p53-responsive enhancer element and a luciferase reporter gene, suggested MCD-1 cells are deficient in wild-type p53 and do not activate p53 on treatment with the anticancer agent adriamycin. The MCD-1 cell line is suggested to represent an abnormally differentiated cell type, which has some properties consistent with a multipotent neuronal phenotype while retaining some properties of immature cells of a glial lineage. The MCD-1 cell line can be used to provide a model of a medulloblastoma cell line that is resistant to growth-controlling and anticancer agents. C1 ST ELIZABETH HOSP,CTR NEUROSCI,SECT PRECLIN NEUROSCI,NEUROPSYCHIAT BRANCH,NIMH,WASHINGTON,DC 20032. NINCDS,LAB MOL MED & NEUROSCI,BETHESDA,MD 20892. GENESYS NEUROSURG ASSOCIATES,FLINT,MI. NR 60 TC 12 Z9 13 U1 0 U2 1 PU HUMANA PRESS INC PI TOTOWA PA 999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 SN 1044-7393 J9 MOL CHEM NEUROPATHOL JI Mol. Chem. Neuropathol. PD OCT-DEC PY 1996 VL 29 IS 2-3 BP 107 EP 126 PG 20 WC Neurosciences; Pathology SC Neurosciences & Neurology; Pathology GA VZ185 UT WOS:A1996VZ18500001 PM 8971690 ER PT J AU Popodi, E Kissinger, JC Andrews, ME Raff, RA AF Popodi, E Kissinger, JC Andrews, ME Raff, RA TI Sea urchin Hox genes: Insights into the ancestral Hox cluster SO MOLECULAR BIOLOGY AND EVOLUTION LA English DT Article DE Heliocidaris erythrogramma; Hox homeobox; echinoderms; sea urchin; ancestral Hox cluster; radial symmetry ID HOMEOBOX GENES; EVOLUTION; EXPRESSION; SEQUENCES; EMBRYOS; ELEGANS AB We describe the Hox cluster in the radially symmetric sea urchin and compare our findings to what is known from clusters in bilaterally symmetric animals. Several Hox genes from the direct-developing sea urchin Heliocidaris erythrogramma are described. CHEF gel analysis shows that the Hox genes are clustered on a less than or equal to 300 kilobase (kb) fragment of DNA, and only a single cluster is present, as in lower chordates and other nonvertebrate metazoans. Phylogenetic analyses of sea urchin, amphioxus, Drosophila, and selected vertebrate Hox genes confirm that the H. erythrogramma genes, and others previously cloned from other sea urchins, belong to anterior, central, and posterior groups. Despite their radial body plan and lack of cephalization, echinoderms retain at least one of the anterior group Hox genes, an orthologue of Hox3. The structure of the echinoderm Hox cluster suggests that the ancestral deuterostome had a Hox cluster more similar to the current chordate cluster than was expected. Sea urchins have at least three Abd-B type genes, suggesting that Abd-B expansion began before the radiation of deuterostomes. C1 NATL INST HLTH,BETHESDA,MD. RP Popodi, E (reprint author), INDIANA UNIV,DEPT BIOL,BLOOMINGTON,IN 47405, USA. RI Kissinger, Jessica/E-9610-2010 OI Kissinger, Jessica/0000-0002-6413-1101 FU NINDS NIH HHS [R01 NS31661] NR 30 TC 55 Z9 57 U1 1 U2 3 PU SOC MOLECULAR BIOLOGY EVOLUTION PI LAWRENCE PA PO BOX 1897, LAWRENCE, KS 66044-8897 SN 0737-4038 J9 MOL BIOL EVOL JI Mol. Biol. Evol. PD OCT PY 1996 VL 13 IS 8 BP 1078 EP 1086 PG 9 WC Biochemistry & Molecular Biology; Evolutionary Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Evolutionary Biology; Genetics & Heredity GA VN652 UT WOS:A1996VN65200003 PM 8865662 ER PT J AU Stolow, MA Bauzon, DD Li, JW Sedgwick, T Liang, VCT Sang, QXA Shi, YB AF Stolow, MA Bauzon, DD Li, JW Sedgwick, T Liang, VCT Sang, QXA Shi, YB TI Identification and characterization of a novel collagenase in Xenopus laevis: Possible roles during frog development SO MOLECULAR BIOLOGY OF THE CELL LA English DT Article ID HUMAN NEUTROPHIL COLLAGENASE; HUMAN FIBROBLAST COLLAGENASE; AMINO-ACID-SEQUENCE; THYROID-HORMONE; HUMAN-SKIN; AMPHIBIAN METAMORPHOSIS; MATRIX METALLOPROTEINASES; EXTRACELLULAR-MATRIX; SUBSTRATE-SPECIFICITY; CLEAVAGE SPECIFICITY AB Matrix metalloproteinases (MMPs) participate in extracellular matrix remodeling and degradation and have been implicated in playing important roles during organ development and pathological processes. Although it has been hypothesized for >30 years that collagenase activities are responsible for collagen degradation during tadpole tail resorption, none of the previously cloned amphibian MMPs have been biochemically demonstrated to be collagenases. Here, we report a novel matrix metalloproteinase gene from metamorphosing Xenopus laevis tadpoles. In vitro biochemical studies demonstrate that this Xenopus enzyme is an interstitial collagenase and has an essentially identical enzymatic activity toward a collagen substrate as the human interstitial collagenase. Sequence comparison of this enzyme to other known MMPs suggests that the Xenopus collagenase is not a homologue of any known collagenases but instead represents a novel collagenase, Xenopus collagenase-4 (xCol4, MMP-18). Interestingly, during development, xCol4; is highly expressed only transiently in whole animals, at approximately the time when tadpole feeding begins, suggesting a role during the maturation of the digestive tract. More improtantly, during metamorphosis, xCol4 is regulated in a tissue-dependent manner. High levels of its mRNA are present as the tadpole tail resorbs. Similarly, its expression is elevated during hindlimb morphogenesis and intestinal remodeling. In addition, when premetamorphic tadpoles are treated with thyroid hormone, the causative agent of metamorphosis, xCol4 expression is induced in the tail. These results suggest that xCol4 may facilitate larval tissue degeneration and adult organogenesis during amphibian metamorphosis. C1 NICHHD,MOL EMBRYOL LAB,NIH,BETHESDA,MD 20892. FLORIDA STATE UNIV,DEPT CHEM,TALLAHASSEE,FL 32306. FLORIDA STATE UNIV,INST MOL BIOPHYS,TALLAHASSEE,FL 32306. NR 60 TC 101 Z9 104 U1 0 U2 1 PU AMER SOC CELL BIOL PI BETHESDA PA PUBL OFFICE 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 1059-1524 J9 MOL BIOL CELL JI Mol. Biol. Cell PD OCT PY 1996 VL 7 IS 10 BP 1471 EP 1483 PG 13 WC Cell Biology SC Cell Biology GA VM880 UT WOS:A1996VM88000001 PM 8898355 ER PT J AU Ramachandra, M Ambudkar, SV Gottesman, MM Pastan, I Hrycyna, CA AF Ramachandra, M Ambudkar, SV Gottesman, MM Pastan, I Hrycyna, CA TI Functional characterization of a glycine 185-to-valine substitution in human P-glycoprotein by using a vaccinia-based transient expression system SO MOLECULAR BIOLOGY OF THE CELL LA English DT Article ID HAMSTER OVARY CELLS; HUMAN MULTIDRUG TRANSPORTER; RESISTANT HUMAN-CELLS; ATPASE ACTIVITY; DRUG TRANSPORT; INSECT CELLS; HUMAN MDR1; MONOCLONAL-ANTIBODIES; CHLORIDE CHANNELS; TUMOR-CELLS AB Human P-glycoprotein (Pgp) is a 170-kDa plasma membrane protein that confers multidrug resistance to otherwise sensitive cells. A mutation in Pgp, G185-->V, originally identified as a spontaneous mutation, was shown previously to alter the drug resistance profiles in cell lines that are stably transfected with the mutant MDR1 cDNA and selected with cytotoxic agents. To understand the mechanism by which the V185 mutation leads to an altered drug resistance profile, we used a transient expression system that eliminates the need for drug selection to attain high expression levels and allows for the rapid characterization of many aspects of Pgp function and biosynthesis. The mutant and wild-type proteins were expressed at similar levels after 24-48 h in human osteosarcoma (HOS) cells by infection with a recombinant vaccinia virus encoding T7 RNA polymerase and simultaneous transfection with a plasmid containing MDR1 cDNA controlled by the T7 promoter. For both mutant and wild-type proteins, photolabeling with [H-3]azidopine and [I-125]iodoarylazidoprazosin, drug-stimulated ATPase activity, efflux of rhodamine 123, and accumulation of radiolabeled vinblastine and colchicine were evaluated. In crude membrane preparations from HOS cells, a higher level of basal Pgp-ATPase activity was observed for the V185 variant than for the wild-type, suggesting partial uncoupling of drug-dependent ATP hydrolysis by the mutant. Several compounds, including verapamil, nicardipine, tetraphenylphosphonium, and prazosin, stimulated ATPase activities of both the wild-type and mutant similarly, whereas cyclosporin A inhibited the ATPase activity of the mutant more efficiently than that of the wild-type. This latter observation explains the enhanced potency of cyclosporin A as an inhibitor of the mutant Pgp. No differences were seen in verapamil-inhibited rhodamine 123 efflux, but the rate of accumulation was slower for colchicine and faster for vinblastine in cells expressing the mutant protein, as compared with those expressing wild-type Pgp. We conclude that the G185-->V mutation confers pleiotropic alterations on Pgp, including an altered basal ATPase activity and altered interaction with substrates and the inhibitor cyclosporin A. C1 NCI,MOL BIOL LAB,NIH,BETHESDA,MD 20892. RP Ramachandra, M (reprint author), NCI,CELL BIOL LAB,DIV BASIC SCI,NIH,BLDG 37,ROOM 1B22,37 CONVENT DR MSC4255,BETHESDA,MD 20892, USA. RI Ambudkar, Suresh/B-5964-2008 NR 51 TC 64 Z9 64 U1 0 U2 0 PU AMER SOC CELL BIOL PI BETHESDA PA PUBL OFFICE 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 1059-1524 J9 MOL BIOL CELL JI Mol. Biol. Cell PD OCT PY 1996 VL 7 IS 10 BP 1485 EP 1498 PG 14 WC Cell Biology SC Cell Biology GA VM880 UT WOS:A1996VM88000002 PM 8898356 ER PT J AU Suh, DS Zhou, YH Ooi, GT Rechler, MM AF Suh, DS Zhou, YH Ooi, GT Rechler, MM TI Dexamethasone stimulation of rat insulin-like growth factor binding protein-1 promoter activity involves multiple cis-elements SO MOLECULAR ENDOCRINOLOGY LA English DT Article ID TYROSINE AMINOTRANSFERASE GENE; STREPTOZOTOCIN-DIABETIC RATS; TRANSCRIPTION FACTOR AP-2; HEPATOMA-CELLS; PHOSPHOENOLPYRUVATE CARBOXYKINASE; GLUCOCORTICOID RESPONSE; HOMEODOMAIN PROTEINS; HORMONE RECEPTORS; IGFBP-1 PROMOTER; SUBUNIT GENE AB Insulin-like growth factor binding protein-1 (IGFBP-1) modulates the mitogenic actions of IGF-I and IGF-II. Dexamethasone increases IGFBP-1 mRNA abundance and gene transcription in rat liver and in H4-II-E rat hepatoma cells. A glucocorticoid response element (GRE) located at nucleotide (nt) -91/-77 is required for dexamethasone to stimulate rat IGFBP-1 promoter activity in transient transfection assays in H4-II-E cells. Mutagenesis of nt -108/-99 (the M4 region of the insulin response element), however, decreased dexamethasone-stimulated promoter activity despite the presence of an intact GRE, suggesting that regulatory sites in addition to the GRE were required for optimal dexamethasone stimulation. To identify these sites, we introduced 5'-deletion and substitution mutations into rat IGFBP-1 promoter fragments coupled to a luciferase reporter gene and transfected these constructs into H4-II-E cells. Three sites are required for optimal basal promoter activity: a site (nt -62/-50) that binds the liver-enriched transcription factor, hepatocyte nuclear factor-1 (HNF-1), the M4 site, and a putative binding site for transcription factor AP-2 (nt -293/-286). The HNF-1 and M4 sites and an upstream site (nt -252/-236) are also involved in dexamethasone stimulation under some, but not all, circumstances. Mutation of either the HNF-1 site or the M4 site decreased dexamethasone stimulation by more than 80% in constructs whose 5'-end was at nt -92, -135, or -235 but not if the 5'-end was at nt -278 or -327. These results suggest that the nt -278/-236 region can compensate for the loss of the HNF-1 site or the M4 site but that the HNF-1 and M4 sites do not compensate for each other in constructs whose 5'-end was at nt -135 or -235, which lack the nt -278/-236 region. The site within the nt -278/-236 region was localized to nt -252/-236 by deoxyribonuclease I protection and transfection assays. Thus, several cis-elements in the rat IGFBP-1 promoter cooperate, in varying combinations, with the low-affinity GRE to allow optimal dexamethasone-stimulated promoter activity. RP Suh, DS (reprint author), NIDDKD, GROWTH & DEV SECT, MOL & CELLULAR ENDOCRINOL BRANCH,NIH,BLDG 10, ROOM 8D14, 10 CTR DR, BETHESDA, MD 20892 USA. NR 53 TC 20 Z9 20 U1 0 U2 3 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0888-8809 J9 MOL ENDOCRINOL JI Mol. Endocrinol. PD OCT PY 1996 VL 10 IS 10 BP 1227 EP 1237 DI 10.1210/me.10.10.1227 PG 11 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VK867 UT WOS:A1996VK86700006 PM 9121490 ER PT J AU McDonald, AH Byrd, LG Mainhart, CR Sopher, J SmithGill, SJ AF McDonald, AH Byrd, LG Mainhart, CR Sopher, J SmithGill, SJ TI Plasmacytoma-refractory BALB/cAnPt mice have naive T cell and highly specific B cell responses to antigen SO MOLECULAR IMMUNOLOGY LA English DT Article DE plasmacytoma; specific pathogen free; B cell repertoire; hen egg lysosome; lymphocyte proliferation; multireactive immunoglobulins ID GERM-FREE; CONVENTIONAL MICE; PATHOGEN-FREE; LYMPHOCYTES; IMMUNOGLOBULIN; GROWTH; ANTIBODIES; ACTIVATION; SECRETION; MOUSE AB Recently, a reduction in the incidence of pristane-induced plasmacytomas in BALB/cAnPt (BALB/c) mice that were kept in viral specific pathogen-free (SPF) conditions has been reported. Environmentally, these SPF-BALB/c mice differed from conventionally-housed (CON) mice only in viral exposure and diet (i.e. sterilization of mouse chow), since microbial colonization of the intestinal tract was seen to be equivalent. This report assessed the ability of SPF- and CON-BALB/c mice to respond to immunologic challenge with soluble antigen, i.e. hen egg white lyzosyme (HEL), as a means of evaluating differences in T and B cell function and, indirectly, evaluating the possible effects these differences might have on plasmacytoma development. When cultured in vitro for 5 days with HEL, HEL-primed lymph node cells (LNC) from SPF-BALB/c mice proliferated to a significantly lesser extent than HEL-primed CON-BALB/c LNC. Moreover, HEL-induced production of IFN-gamma and IL-5 was significantly lower in SPF LNC. Serum IgG1 levels were 10-fold lower in SPF-BALB/c mice with, or without prior immunization with HEL and were not reconstituted by repeated injections of HEL in adjuvant. Serum IgM levels of SPF- and CON-BALB/c mice were equivalent. This reduction in immune responses could not be attributed to a lack of colonization of secondary lymphoid organs, since flow cytometric analysis of LNC revealed no difference in the number of recoverable cells and the proportion of lymphocyte subsets (CD4(+), CD8(+) and CD45(+) cells) obtained from SPF- and CON-BALB/c mice. However, only CON LNC were induced to increase surface expression of CD44 after antigenic or mitogenic stimulation in vitro. Antibody responsiveness to HEL, as evidenced by serum anti-HEL binding or splenic hybridoma studies, demonstrated higher levels of IgG1 antibodies in CON-BALB/c mice than in SPF mice. However, a greater proportion of the SPF IgG1 antibodies present were specifically directed against HEL, so that specific activity was greater in SPF-BALB/c mice. Therefore, while SPF-BALB/c mice have a more restricted response to HEL than CON-BALB/c mice, those antibodies that are produced are more specifically directed against HEL with very little apparent bystander/polyclonal activation of multireactive cells. Resistance to plasmacytomas in SPF-BALB/c mice, therefore, may stem from a reduced number of circulating memory T and B cells, which are capable of reacting and/or crossreacting with a chronic inflammatory stimulus. (C) 1997 Elsevier Science Ltd. C1 VIRION SYST,ROCKVILLE,MD 20850. NCI,GENET LAB,NIH,BETHESDA,MD 20892. RP McDonald, AH (reprint author), MED COLL WISCONSIN,DEPT PATHOL,MILWAUKEE,WI 53226, USA. NR 49 TC 2 Z9 2 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, ENGLAND OX5 1GB SN 0161-5890 J9 MOL IMMUNOL JI Mol. Immunol. PD OCT PY 1996 VL 33 IS 15 BP 1183 EP 1196 DI 10.1016/S0161-5890(96)00103-4 PG 14 WC Biochemistry & Molecular Biology; Immunology SC Biochemistry & Molecular Biology; Immunology GA WN003 UT WOS:A1996WN00300005 PM 9070667 ER PT J AU Brinkmann, U AF Brinkmann, U TI Recombinant immunotoxins: Protein engineering for cancer therapy SO MOLECULAR MEDICINE TODAY LA English DT Review ID SINGLE-CHAIN IMMUNOTOXINS; STABILIZED FV FRAGMENT; PSEUDOMONAS-EXOTOXIN; ANTITUMOR-ACTIVITY; CONTINUOUS-INFUSION; ESCHERICHIA-COLI; DIPHTHERIA-TOXIN; FUSION PROTEIN; MICE; BINDING AB Recombinant immunotoxins for cancer therapy are composed of the variable regions of 'cancer-specific' antibodies fused to truncated toxins that are usually derived from bacteria or plants, Protein engineering has been used to modify these molecules so that the toxin moiety by itself does not bind to normal human cells, but retains all other cytotoxic functions, The antibody moiety directs the toxin selectively to cancer cells, which are killed; cells that do not carry that particular cancer antigen are not recognized and are therefore spared, Many recombinant immunotoxins show a high degree of cytotoxic activity and specificity towards cancer cells cultured in vitro and have been shown to cause the regression of human tumor xenografts grown in mice. Clinical trials that are in progress will show whether these promising pre-clinical results can be translated into successful cancer therapy. RP Brinkmann, U (reprint author), NCI,MOL BIOL LAB,NIH,DIV BASIC SCI,BLDG 37,ROOM 4B20,37 CONVENT DR,MSC 4255,BETHESDA,MD 20892, USA. NR 42 TC 17 Z9 17 U1 0 U2 2 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, OXON, ENGLAND OX5 1GB SN 1357-4310 J9 MOL MED TODAY JI Mol. Med. Today PD OCT PY 1996 VL 2 IS 10 BP 439 EP 446 DI 10.1016/1357-4310(96)84848-9 PG 8 WC Biochemistry & Molecular Biology; Cell Biology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Cell Biology; Research & Experimental Medicine GA VN501 UT WOS:A1996VN50100010 PM 8897439 ER PT J AU Heinzen, RA Howe, D Mallavia, LP Rockey, DD Hackstadt, T AF Heinzen, RA Howe, D Mallavia, LP Rockey, DD Hackstadt, T TI Developmentally regulated synthesis of an unusually small, basic peptide by Coxiella burnetii SO MOLECULAR MICROBIOLOGY LA English DT Article ID BACTERIAL GENE-EXPRESSION; OUTER-MEMBRANE PROTEIN; BACILLUS-SUBTILIS; ESCHERICHIA-COLI; CHLAMYDIA-TRACHOMATIS; DNA-BINDING; CLONING; PH; SPORULATION; SEQUENCE AB Coxiella burnetii undergoes a poorly defined developmental cycle within phagolysosomes of eukaryotic host cells. Two distinct developmental forms are part of this cycle: a small-cell variant (SCV) and large-cell variant (LCV). Ultrastructurally, the SCV is distinguished from the LCV by its smaller size and condensed chromatin. At a molecular level, little is known about morphogenesis in C. burnetii, and no proteins specific to the SCV have been identified. Preparative isoelectric focusing was conducted to purify basic proteins possibly involved in SCV chromatin structure. A predominant protein of low M(r) was present in the most basic fraction, eluting with a pH of approx. 11. Degenerate deoxyoligonucleotides corresponding to the N-terminal sequence of this protein were used to recover a cosmid clone from a C. burnetii genomic library. Nucleotide sequencing of insert DNA revealed an open reading frame designated scvA (small-cell-variant protein A) with coding potential for a 30 amino acid protein (ScvA) with a predicted M(r) of 3610. ScvA is 46% arginine plus 46% glutamine with a predicted pi of 12.6. SDS-PAGE and silver staining of lysates of SCV and LCV purified by caesium chloride-equilibrium density centrifugation revealed a number of proteins unique to each cell type. Immunoblot analysis with ScvA antiserum demonstrated the presence of ScvA only in the SCV. By immunoelectron microscopy, ScvA antiserum labelled only the SCV, with the label concentrated on the condensed nucleoid. In addition, ScvA bound double-stranded DNA in gel mobility-shift assays. A 66% reduction in the mean number of gold particles per Coxiella cell was observed at 12 h post-infection when compared with the starting inoculum. Collectively, these data suggest that synthesis of ScvA is developmentally regulated, and that the protein may serve a structural or functional role as an integral component of the SCV chromatin. Moreover, degradation of this protein may be a necessary prerequisite for morphogenesis from SCV to LCV. C1 WASHINGTON STATE UNIV,DEPT MICROBIOL,PULLMAN,WA 99164. RP Heinzen, RA (reprint author), NIAID,ROCKY MT LABS,INTRACELLULAR PARASITES LAB,NIH,HAMILTON,MT 59840, USA. FU NIAID NIH HHS [AI20190] NR 49 TC 31 Z9 32 U1 0 U2 1 PU BLACKWELL SCIENCE LTD PI OXFORD PA OSNEY MEAD, OXFORD, OXON, ENGLAND OX2 0EL SN 0950-382X J9 MOL MICROBIOL JI Mol. Microbiol. PD OCT PY 1996 VL 22 IS 1 BP 9 EP 19 DI 10.1111/j.1365-2958.1996.tb02651.x PG 11 WC Biochemistry & Molecular Biology; Microbiology SC Biochemistry & Molecular Biology; Microbiology GA VM856 UT WOS:A1996VM85600002 PM 8899704 ER PT J AU Valdivia, RH Falkow, S AF Valdivia, RH Falkow, S TI Bacterial genetics by flow cytometry: Rapid isolation of Salmonella typhimurium acid-inducible promoters by differential fluorescence induction SO MOLECULAR MICROBIOLOGY LA English DT Article ID MULTIPLE ANTIBIOTIC-RESISTANCE; PENICILLIN-BINDING PROTEIN-2; ESCHERICHIA-COLI; TOLERANCE RESPONSE; MULTIDRUG-RESISTANCE; NUCLEOTIDE-SEQUENCE; SHOCK PROTEINS; CHAIN-LENGTH; PBPA GENE; RNASE-I AB The ability of Salmonella typhimurium to survive and replicate within murine macrophages is dependent on a low phagosomal pH. This requirement for an acidic vacuole suggests that low pH is an important environmental stimulus for the transcription of genes necessary for intracellular survival. To study the behaviour of acid-inducible genes in response to the phagosomal environment, we have applied a novel enrichment strategy, termed (d) under bar ifferential (f) under bar luorescence induction (DFI), to screen an S. typhimurium library for promoters that are upregulated at pH 4.5. DFI utilizes a fluorescence-enhanced green fluorescent protein (GFP) and a fluorescence-activated cell sorter (FACS) to perform genetic selection. In the presence of an inducing stimulus, such as low pH, a FACS is used to sort highly fluorescent bacterial clones bearing random promoters fused to the mutant GFP protein (GFPmut). This population is then amplified at neutral pH and the least fluorescent population is sorted. Sequential sorts for fluorescent and nonfluorescent bacteria in the presence or absence of inducing conditions rapidly enriches for promoter fusions that are regulated by the inducing stimulus. We have identified eight acid-inducible promoters and quantified their expression in response to pH 4.5 and to the phagosome milieu. These acid-inducible promoters exhibited extensive homology to promoter regions of genes encoding for cell-surface-maintenance enzymes, stress proteins, and generalized efflux pumps. Only a subset of these promoters was induced in macrophages with kinetics and levels of expression that do not necessarily correlate with in vitro pH-shock induction. This suggests that while low pH is a relevant inducer of intracellular gene expression, additional stimuli in the macrophage can modulate the expression of acid-inducible genes. C1 NIAID, ROCKY MT LABS, HAMILTON, MT 59840 USA. RP STANFORD UNIV, SCH MED, DEPT IMMUNOL & MICROBIOL, STANFORD, CA 94305 USA. FU NIAID NIH HHS [AI 26195]; NIDDK NIH HHS [DK 38707] NR 46 TC 303 Z9 319 U1 1 U2 22 PU WILEY-BLACKWELL PI HOBOKEN PA 111 RIVER ST, HOBOKEN 07030-5774, NJ USA SN 0950-382X EI 1365-2958 J9 MOL MICROBIOL JI Mol. Microbiol. PD OCT PY 1996 VL 22 IS 2 BP 367 EP 378 DI 10.1046/j.1365-2958.1996.00120.x PG 12 WC Biochemistry & Molecular Biology; Microbiology SC Biochemistry & Molecular Biology; Microbiology GA VP097 UT WOS:A1996VP09700016 PM 8930920 ER PT J AU Liu, C Zhuo, XL Gonzalez, FJ Ding, XX AF Liu, C Zhuo, XL Gonzalez, FJ Ding, XX TI Baculovirus-Mediated expression and characterization of rat CYP2A3 and human CYP2A6: Role in metabolic activation of nasal toxicants SO MOLECULAR PHARMACOLOGY LA English DT Article ID STEROID 15-ALPHA-HYDROXYLASE; CYTOCHROME-P-450 ISOZYME; COUMARIN 7-HYDROXYLATION; II P-45015-ALPHA; RABBIT; MICROSOMES; SEQUENCE; CDNA; GENE; IDENTIFICATION AB Cytochrome P450 2A3 (CYP2A3) was previously identified in rat lung by cDNA cloning and recently found to be expressed at a high level in the olfactory mucosa. In the current study, CYP2A3 was expressed in insect cells lacking endogenous cytochrome P450 (P450) activity, and the substrate specificity of the recombinant cytochrome was characterized and compared with that of CYP2A6, a human ortholog of rat CYP2A3, which has been detected in human olfactory mucosa as well as in liver. The CYP2A3 and CYP2A6 cDNAs were cloned into baculovirus, and recombinant viruses were used to produce active enzymes in Spodoptera frugiperta (SF9) cells. The metabolic activities of S. frugiperta cell microsomal fractions containing CYP2A3 or CYP2A6 were studied in a reconstituted system with purified rabbit NADPH-P450 reductase. CYP2A3 was found to be active toward testosterone, producing 15 alpha-hydroxytestosterone and several other metabolites, but it had only low activity toward coumarin. On the other hand, CYP2A6 was active toward coumarin but not toward testosterone. However, both enzymes were active in the metabolic activation of hexamethylphosphoramide, a nasal procarcinogen, and 2,6-dichlorobenzonitrile (DCBN), a herbicide known to cause tissue-specific toxicity in the olfactory mucosa of rodents at very low doses. In addition, both enzymes were active toward 4-nitrophenol, a preferred substrate for CYP2E1. Consistent with CYP2A3 being a major catalyst in microsomal metabolism of DCBN, the activities of both CYP2A3 and rat olfactory microsomes in DCBN metabolism were inhibited strongly by metyrapone and methoxsalen (ID50 <1 mu M, with DCBN at 30 mu M), but only marginally by 4-methylpyrazole, an inhibitor of CYP2E1. In contrast, the activity of CYP2A6 was only weakly inhibited by metyrapone or methoxsalen (ID50 >50 mu M). Thus, rat CYP2A3 and human CYP2A6 have differences in substrate specificity as well as tissue distribution. These findings should be taken into account when assessing the risk of exposure to potential nasal toxicants in humans. C1 NEW YORK STATE DEPT HLTH,WADSWORTH CTR LABS & RES,LAB HUMAN TOXICOL & MOL EPIDEMIOL,ALBANY,NY 12201. SUNY ALBANY,SCH PUBL HLTH,DEPT ENVIRONM HLTH & TOXICOL,ALBANY,NY 12201. NCI,MOL CARCINOGENESIS LAB,NATL INST HLTH,BETHESDA,MD 20892. FU NIEHS NIH HHS [ES07462] NR 38 TC 69 Z9 70 U1 0 U2 3 PU WILLIAMS & WILKINS PI BALTIMORE PA 351 WEST CAMDEN ST, BALTIMORE, MD 21201-2436 SN 0026-895X J9 MOL PHARMACOL JI Mol. Pharmacol. PD OCT PY 1996 VL 50 IS 4 BP 781 EP 788 PG 8 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA VN831 UT WOS:A1996VN83100011 PM 8863822 ER PT J AU Yu, DH Zhang, L Eisele, JL Bertrand, D Changeux, JP Weight, FF AF Yu, DH Zhang, L Eisele, JL Bertrand, D Changeux, JP Weight, FF TI Ethanol inhibition of nicotinic acetylcholine type alpha 7 receptors involves the amino-terminal domain of the receptor SO MOLECULAR PHARMACOLOGY LA English DT Article ID GATED ION-CHANNEL; POTASSIUM CHANNEL; LIGAND-BINDING; ALCOHOLS; POTENTIATION; ANESTHETICS; ENDPLATE; PROTEIN; OOCYTES; CLONING AB Recent studies have suggested that alcohols can affect the function of neuro transmitter-gated ion channels by a direct interaction with the receptor protein. However, the molecular region of the receptor protein that mediates the alcohol action is not known. To address this question, we studied the effect of ethanol on the function of recombinant nicotinic acetylcholine type alpha 7 (nACh(alpha 7)) receptors, 5-hydroxytryptamine (serotonin) type 3 (5-HT3) receptors, and a chimeric receptor constructed from these two receptors. The receptors were expressed in Xenopus oocytes and their function was studied using the two-electrode voltage-clamp technique. Ethanol inhibited the response of nACh(alpha 7) receptors in a concentration-dependent manner over the concentration range of 5-100 mM; the EC(50) for this inhibition was 33 mM ethanol. Ethanol decreased the maximal amplitude (E(max)) of the nACh(alpha 7) receptor agonist concentration-response curve, without significantly affecting the EC(50). In contrast, ethanol potentiated 5-HT3 receptor-mediated responses at low agonist concentrations. The potentiation was concentration-dependent over the concentration range of 10-100 mM; the EC(50) for this potentiation was 57 mM ethanol. The magnitude of the ethanol potentiation of 5-HT3 receptor-mediated responses decreased with increasing agonist concentration. The chimeric receptor had the amino-terminal domain from the nACh(alpha 7) receptor and the transmembrane and carboxyl-terminal domains from the 5-HT3 receptor. Ethanol was found to inhibit the function of this chimeric receptor in a manner similar to that of nACh(alpha 7) receptors. Because the inhibition transfers with the amino-terminal domain of the receptor, the observations suggest that the amino-terminal domain of the receptor is involved in the inhibition. C1 NIAAA,LMCN,NIH,ROCKVILLE,MD 20852. INST PASTEUR,UR D1284 CNRS,F-75724 PARIS 15,FRANCE. CTR MED UNIV GENEVA,DEPT PHYSIOL,FAC MED,CH-1211 GENEVA 4,SWITZERLAND. NR 35 TC 87 Z9 88 U1 0 U2 0 PU WILLIAMS & WILKINS PI BALTIMORE PA 351 WEST CAMDEN ST, BALTIMORE, MD 21201-2436 SN 0026-895X J9 MOL PHARMACOL JI Mol. Pharmacol. PD OCT PY 1996 VL 50 IS 4 BP 1010 EP 1016 PG 7 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA VN831 UT WOS:A1996VN83100037 PM 8863848 ER PT J AU Martinez, RA AF Martinez, RA TI National Institute of Mental Health: An update on women and minorities in research SO MOUNT SINAI JOURNAL OF MEDICINE LA English DT Article AB Guidelines for the recruitment and retention of women and minorities as subjects in clinical trials have been developed at the National Institutes of Health. Training more women and minority investigators is also a National Institutes of Health priority. These individuals, like all other applicants, however, must be sophisticated consumers of the federal grants process; This article presents information about administration changes for entry-level research awards to help potential applicants understand the funding process. RP Martinez, RA (reprint author), NIMH,GERIATR PSYCHIAT RES PROGRAM,MENTAL DISORDERS AGING RES BRANCH,5600 FISHERS LANE,ROCKVILLE,MD 20857, USA. NR 0 TC 1 Z9 1 U1 1 U2 3 PU MOUNT SINAI HOSPITAL PI NEW YORK PA BOX 1094 ONE GUSTAVE L LEVY PLACE ATTN: CIRCULATION ASST, NEW YORK, NY 10029-6574 SN 0027-2507 J9 MT SINAI J MED JI Mt. Sinai J. Med. PD OCT-NOV PY 1996 VL 63 IS 5-6 BP 332 EP 334 PG 3 WC Medicine, General & Internal SC General & Internal Medicine GA VK588 UT WOS:A1996VK58800011 PM 8898538 ER PT J AU Rodbell, M AF Rodbell, M TI G proteins: Out of the cytoskeletal closet SO MOUNT SINAI JOURNAL OF MEDICINE LA English DT Article ID BETA-GAMMA-SUBUNITS; NUCLEOSIDE DIPHOSPHATE KINASE; NUCLEOTIDE REGULATORY PROTEIN; STIMULATORY G-PROTEIN; GTP-BINDING PROTEINS; SIGNAL-TRANSDUCTION; ADENYLYL CYCLASE; ADRENERGIC-RECEPTOR; ALPHA-SUBUNIT; GUANINE-NUCLEOTIDES AB This article contains a brief review of GTP-binding proteins (G protein) signaling mechanism with emphasis on accumulated information which suggests that G proteins are multimeric proteins structured such that one receptor can catalytically activate each of the monomers as it moves in an oscillatory fashion along the multimeric chain. Movement is dictated by the binding of GTP or GDP controlled by the exchange reaction induced by agonist binding to the receptor. Based on the dynamic instability model for the interactions of myosin and F-actin, the hypothesis is presented that a GTP-bound monomer is released from one end of the multimer allowing it to interact with effecters such as adenylyl cyclase embedded in the plasma membrane. Association with the enzyme results in a transition, state of the enzyme-G protein complex. In the presence of magnesium, the GTPase on the alpha-subunit of Gs (which stimulates adenylyl cyclase) is activated, resulting in the interaction of the alpha- and beta gamma-subunits of Gs with different domains of adenylyl cyclase. In this fashion, the GTPase is not simply a turnoff mechanism, but induces a new configuration of the G protein-cyclase complex allowing for greatly enhanced cyclic AMP formation. Dissociation of bound Pi may be one of the rate-limiting steps allowing for cycling of G proteins between the multimer-receptor complex and adenylyl cyclase. RP Rodbell, M (reprint author), NIEHS,SIGNAL TRANSDUCT SECT,CELLULAR & MOL PHARMACOL LAB,BLDG 18-01,RES TRIANGLE PK,NC 27709, USA. NR 62 TC 3 Z9 3 U1 1 U2 2 PU MOUNT SINAI HOSPITAL PI NEW YORK PA BOX 1094 ONE GUSTAVE L LEVY PLACE ATTN: CIRCULATION ASST, NEW YORK, NY 10029-6574 SN 0027-2507 J9 MT SINAI J MED JI Mt. Sinai J. Med. PD OCT-NOV PY 1996 VL 63 IS 5-6 BP 381 EP 386 PG 6 WC Medicine, General & Internal SC General & Internal Medicine GA VK588 UT WOS:A1996VK58800018 PM 8898544 ER PT J AU Spector, SA Gordon, PL Feuerstein, IM Sivakumar, K Hurley, BF Dalakas, MC AF Spector, SA Gordon, PL Feuerstein, IM Sivakumar, K Hurley, BF Dalakas, MC TI Strength gains without muscle injury after strength training in patients with postpolio muscular atrophy SO MUSCLE & NERVE LA English DT Article DE postpolio syndrome; exercise; muscle injury ID ELBOW FLEXOR MUSCLES; NEUROMUSCULAR SYMPTOMS; RESISTANCE EXERCISE; SKELETAL-MUSCLE; CREATINE-KINASE; MOTOR UNIT; POLIOMYELITIS; CONTRACTIONS; COORDINATION; HYPERTROPHY AB We evaluated changes in the dynamic and isometric strength in the newly weakened quadriceps muscles and asymptomatic triceps muscles of 6 patients with postpolio muscular atrophy (PPMA) after 10 weeks of progressive resistance strength training, Alterations in muscle size were determined with magnetic resonance imaging, Serum creatine kinase levels were measured throughout training, and histological signs of muscle injury and changes in muscle fiber size and types were assessed with muscle biopsies before and after training, Exercise training led to an increase in dynamic strength of 41% and 61% for the two knee extensor tests, and 54% and 71% for the two elbow extensor tests. Up to 20% of the improvement was maintained 5 months after cessation of training, Isometric strength, whole muscle cross-sectional areas of quadriceps and triceps muscles, and serum muscle enzymes did not change, No destructive histopathological changes were noted in the repeat muscle biopsies, and no consistent changes in muscle fiber size or fiber type percentages were observed. These results demonstrate that a supervised resistance training program can lead to significant gains in dynamic strength of both symptomatic and asymptomatic muscles of PPMA patients without serological or histological evidence of muscular damage, (C) 1996 John Wiley & Sons, Inc. C1 NINCDS,NEUROMUSCULAR DIS SECT,NIH,BETHESDA,MD 20892. NIH,CTR CLIN,DEPT RADIOL,BETHESDA,MD 20892. UNIV MARYLAND,DEPT KINESIOL,EXERCISE SCI LAB,COLLEGE PK,MD 20742. NR 37 TC 24 Z9 25 U1 0 U2 1 PU JOHN WILEY & SONS INC PI NEW YORK PA 605 THIRD AVE, NEW YORK, NY 10158-0012 SN 0148-639X J9 MUSCLE NERVE JI Muscle Nerve PD OCT PY 1996 VL 19 IS 10 BP 1282 EP 1290 DI 10.1002/(SICI)1097-4598(199610)19:10<1282::AID-MUS5>3.0.CO;2-A PG 9 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA VH390 UT WOS:A1996VH39000006 PM 8808654 ER PT J AU Reiter, Y Brinkmann, U Lee, BK Pastan, I AF Reiter, Y Brinkmann, U Lee, BK Pastan, I TI Engineering antibody Fv fragments for cancer detection and therapy: Disulfide-stabilized Fv fragments SO NATURE BIOTECHNOLOGY LA English DT Review DE dsFv; scFv; antibody engineering; immunotoxin ID SINGLE-CHAIN FV; T-CELL RECEPTOR; CONSERVED FRAMEWORK REGIONS; PSEUDOMONAS EXOTOXIN; RECOMBINANT IMMUNOTOXIN; ESCHERICHIA-COLI; INTERLEUKIN-2 RECEPTOR; MONOCLONAL-ANTIBODIES; ANTITUMOR-ACTIVITY; DIPHTHERIA-TOXIN AB Disulfide-stabilized Fv fragments of antibodies (dsFv) are molecules in which the V-H-V-L heterodimer is stabilized by an interchain disulfide bond engineered between structurally conserved framework positions distant from complementarity-determining regions (CDRs). This method of stabilization is applicable for the stabilization of many antibody Fvs and has also been applied to a T-cell receptor Fv, A summary of the design strategy, and the construction and production of various dsFvs and dsFv-fusion proteins is presented, Included in the discussion are the biochemical features of dsFvs in comparison with scFvs, the effect of disulfide stabilization on Fv binding and activity, and various applications of dsFvs and dsFv-immunotoxins for tumor imaging and the treatment of solid tumors in animal models. C1 NCI, MOL BIOL LAB, DIV BASIC SCI, NIH, BETHESDA, MD 20892 USA. RI Lee, Byungkook/E-4564-2011 OI Lee, Byungkook/0000-0002-3339-4582 NR 74 TC 111 Z9 128 U1 0 U2 9 PU NATURE PUBLISHING GROUP PI NEW YORK PA 75 VARICK ST, 9TH FLR, NEW YORK, NY 10013-1917 USA SN 1087-0156 EI 1546-1696 J9 NAT BIOTECHNOL JI Nat. Biotechnol. PD OCT PY 1996 VL 14 IS 10 BP 1239 EP 1245 DI 10.1038/nbt1096-1239 PG 7 WC Biotechnology & Applied Microbiology SC Biotechnology & Applied Microbiology GA VK618 UT WOS:A1996VK61800020 PM 9631086 ER PT J AU Szkudlinski, MW Teh, NG Grossmann, M Tropea, JE Weintraub, BD AF Szkudlinski, MW Teh, NG Grossmann, M Tropea, JE Weintraub, BD TI Engineering human glycoprotein hormone superactive analogues SO NATURE BIOTECHNOLOGY LA English DT Article DE hTSH; hCG; analogue design ID HUMAN CHORIONIC-GONADOTROPIN; RECOMBINANT HUMAN THYROTROPIN; FOLLICLE-STIMULATING-HORMONE; ALPHA-SUBUNIT; BETA-SUBUNIT; CHORIOGONADOTROPIN RECEPTOR; METABOLIC-CLEARANCE; TERMINAL RESIDUES; BIOACTIVITY; FOLLITROPIN AB We report the generation of superactive analogues of human glycoprotein hormones, with potential applications in thyroid and reproductive disorders. Current biological and structural data were used to rationalize mutagenesis. The 11-20 region in the alpha-subunit with a cluster of lysine residues forms a previously unrecognized domain critical for receptor binding and signal transduction, as well as an important motif in the evolution of glycoprotein hormone activities. The gradual elimination of basic residues in the cu-subunit coincided with the evolutionary divergence of the hominids from the Old World monkeys. By selective reconstitution of certain critical residues present in homologous nonhuman hormones we have developed human thyroid stimulating hormone and chorionic gonadotropin analogues with substantial increases in receptor binding affinity and bioactivity, thus providing a paradigm for the design of novel therapeutic protein analogues. C1 NIDDKD, MOL & CELLULAR ENDOCRINOL BRANCH, NIH, BETHESDA, MD 20892 USA. RP UNIV MARYLAND, SCH MED,DEPT MED,INST HUMAN VIROL, LAB MOL ENDOCRINOL, 725 W LOMBARD ST, BALTIMORE, MD 21201 USA. NR 44 TC 77 Z9 77 U1 0 U2 3 PU NATURE PUBLISHING GROUP PI NEW YORK PA 75 VARICK ST, 9TH FLR, NEW YORK, NY 10013-1917 USA SN 1087-0156 EI 1546-1696 J9 NAT BIOTECHNOL JI Nat. Biotechnol. PD OCT PY 1996 VL 14 IS 10 BP 1257 EP 1263 DI 10.1038/nbt1096-1257 PG 7 WC Biotechnology & Applied Microbiology SC Biotechnology & Applied Microbiology GA VK618 UT WOS:A1996VK61800023 PM 9631089 ER PT J AU Hamer, DH AF Hamer, DH TI The heritability of happiness SO NATURE GENETICS LA English DT Editorial Material ID TRAIT RP Hamer, DH (reprint author), NCI,NATL INST HLTH,BIOCHEM LAB,BLDG 37,RM 4A13,37 CONVENT DR,BETHESDA,MD 20892, USA. NR 9 TC 34 Z9 35 U1 0 U2 4 PU NATURE PUBLISHING CO PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 SN 1061-4036 J9 NAT GENET JI Nature Genet. PD OCT PY 1996 VL 14 IS 2 BP 125 EP 126 DI 10.1038/ng1096-125 PG 2 WC Genetics & Heredity SC Genetics & Heredity GA VL446 UT WOS:A1996VL44600005 PM 8841176 ER PT J AU Bellus, GA Gaudenz, K Zackai, EH Clarke, LA Szabo, J Francomano, CA Muenke, M AF Bellus, GA Gaudenz, K Zackai, EH Clarke, LA Szabo, J Francomano, CA Muenke, M TI Identical mutations in three different fibroblast growth factor receptor genes in autosomal dominant craniosynostosis syndromes SO NATURE GENETICS LA English DT Letter ID PFEIFFER-SYNDROME; CROUZON-SYNDROME; ACHONDROPLASIA; DOMAIN C1 UNIV PENN,SCH MED,CHILDRENS HOSP PHILADELPHIA,DEPT PEDIAT,DIV HUMAN GENET & MOL BIOL,PHILADELPHIA,PA 19104. UNIV PENN,SCH MED,CHILDRENS HOSP PHILADELPHIA,DEPT GENET,PHILADELPHIA,PA 19104. NIH,MED GENET BRANCH,NATL CTR HUMAN GENOME RES,BETHESDA,MD 20892. JOHNS HOPKINS UNIV,SCH MED,CTR MED GENET,BALTIMORE,MD 21287. BRITISH COLUMBIS INST CHILD & FAMILY HLTH,DEPT MED GENET,VANCOUVER,BC V5Z 4H4,CANADA. OI Clarke, Lorne/0000-0003-0512-7281 FU NICHD NIH HHS [R29 HD28732, R01 HD29862] NR 27 TC 228 Z9 230 U1 0 U2 7 PU NATURE PUBLISHING CO PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 SN 1061-4036 J9 NAT GENET JI Nature Genet. PD OCT PY 1996 VL 14 IS 2 BP 174 EP 176 DI 10.1038/ng1096-174 PG 3 WC Genetics & Heredity SC Genetics & Heredity GA VL446 UT WOS:A1996VL44600022 PM 8841188 ER PT J AU Oddoux, C Struewing, JP Clayton, CM Neuhausen, S Brody, LC Kaback, M Haas, B Norton, L Borgen, P Jhanwar, S Goldgar, D Ostrer, H Offit, K AF Oddoux, C Struewing, JP Clayton, CM Neuhausen, S Brody, LC Kaback, M Haas, B Norton, L Borgen, P Jhanwar, S Goldgar, D Ostrer, H Offit, K TI The carrier frequency of the BRCA2 6174delT mutation among Ashkenazi Jewish individuals is approximately 1% SO NATURE GENETICS LA English DT Letter ID CANCER SUSCEPTIBILITY GENE; OVARIAN-CANCER; BREAST-CANCER; FAMILIES C1 NYU,MED CTR,DEPT PEDIAT,HUMAN GENET PROGRAM,NEW YORK,NY 10016. NCI,GENET EPIDEMIOL BRANCH,NIH,BETHESDA,MD 20892. UNIV UTAH,SCH MED,DEPT MED INFORMAT,GENET EPIDEMIOL GRP,SALT LAKE CITY,UT 84108. NIH,LAB GENE TRANSFER,NATL CTR HUMAN GENOME RES,BETHESDA,MD 20892. UNIV CALIF SAN DIEGO,DEPT PEDIAT & REPROD MED,SAN DIEGO,CA 92123. MEM SLOAN KETTERING CANC CTR,DEPT HUMAN GENET,NEW YORK,NY 10021. MEM SLOAN KETTERING CANC CTR,DEPT MED,BREAST CANC MED SERV,NEW YORK,NY 10021. MEM SLOAN KETTERING CANC CTR,DEPT SURG,BREAST SERV,NEW YORK,NY 10021. INT AGCY RES CANC,UNIT GENET EPIDEMIOL,LYON 08,FRANCE. RI Struewing, Jeffery/C-3221-2008; Struewing, Jeffery/I-7502-2013 OI Struewing, Jeffery/0000-0002-4848-3334 FU NCI NIH HHS [P20 CA 58232, R01 CA 55914, R01 CA 65673] NR 16 TC 263 Z9 265 U1 0 U2 8 PU NATURE PUBLISHING CO PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 SN 1061-4036 J9 NAT GENET JI Nature Genet. PD OCT PY 1996 VL 14 IS 2 BP 188 EP 190 DI 10.1038/ng1096-188 PG 3 WC Genetics & Heredity SC Genetics & Heredity GA VL446 UT WOS:A1996VL44600026 PM 8841192 ER PT J AU Donovan, MJ Hahn, R Tessarollo, L Hempstead, BL AF Donovan, MJ Hahn, R Tessarollo, L Hempstead, BL TI Identification of an essential nonneuronal function of neurotrophin 3 in mammalian cardiac development SO NATURE GENETICS LA English DT Letter ID NEURAL CREST CELLS; NERVOUS-SYSTEM; TARGETED DISRUPTION; MESSENGER-RNA; TRKC; RECEPTORS; EXPRESSION; HEART; GENE; RAT C1 NCI,FREDERICK CANC RES & DEV CTR,NEURAL DEV GRP,ABL BASIC RES PROGRAM,FREDERICK,MD 21702. CHILDRENS HOSP,DEPT PATHOL,BOSTON,MA 02115. CORNELL UNIV,COLL MED,DEPT MED,NEW YORK,NY 10021. FU NINDS NIH HHS [NS30687] NR 25 TC 122 Z9 129 U1 0 U2 2 PU NATURE PUBLISHING CO PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 SN 1061-4036 J9 NAT GENET JI Nature Genet. PD OCT PY 1996 VL 14 IS 2 BP 210 EP 213 DI 10.1038/ng1096-210 PG 4 WC Genetics & Heredity SC Genetics & Heredity GA VL446 UT WOS:A1996VL44600032 PM 8841198 ER PT J AU Marx, PA Spira, AI Gettie, A Dailey, PJ Veazey, RS Lackner, AA Mahoney, CJ Miller, CJ Claypool, LE Ho, DD Alexander, NJ AF Marx, PA Spira, AI Gettie, A Dailey, PJ Veazey, RS Lackner, AA Mahoney, CJ Miller, CJ Claypool, LE Ho, DD Alexander, NJ TI Progesterone implants enhance SIV vaginal transmission and early virus load SO NATURE MEDICINE LA English DT Article ID SIMIAN IMMUNODEFICIENCY VIRUS; INTRAVAGINAL INOCULATION; RHESUS MACAQUES; INFECTION; TYPE-2; MODEL; CYCLE; HIV AB Simian immunodeficiency virus (SIV) can cross the intact vaginal epithelium to establish a systemic infection in macaques (mac). Using this SIVmac model, we found that subcutaneous progesterone implants, which could mimic hormonally based contraceptives, thinned the vaginal epithelium and enhanced SIV vaginal transmission 7.7-fold over that observed in macaques treated with placebo implants and exposed to SIV in the follicular phase of the menstrual cycle. Progesterone treatment also increased the number of SIV DNA-positive cells in the vaginal lamina propria as detected by in situ polymerase chain reaction analysis. Moreover, plasma viral RNA was elevated for the first three months in macaques with progesterone implants, and three of the progesterone-treated macaques developed relatively rapid disease courses. This study shows that SIV genital infection and disease course are enhanced by subcutaneous implants containing progesterone when compared with the rate of vaginal transmission in the follicular phase. C1 NYU,SCH MED,DEPT MICROBIOL,NEW YORK,NY 10016. CHIRON CORP,NUCLE ACID SYST,EMERYVILLE,CA 94608. HARVARD UNIV,NEW ENGLAND REG PRIMATE RES CTR,SCH MED,SOUTHBOROUGH,MA 01772. NYU,MED CTR,LAB EXPT MED & SURG PRIMATES,TUXEDO PK,NY 10987. UNIV CALIF DAVIS,CALIF REG PRIMATE RES CTR,DAVIS,CA 95616. EASTERN VIRGINIA MED SCH,CONRAD PROGRAM,ARLINGTON,VA 22209. NICHHD,CONTRACEPT DEV BRANCH,NIH,BETHESDA,MD 20892. RP Marx, PA (reprint author), ROCKEFELLER UNIV,AARON DIAMOND AIDS RES CTR,455 1ST AVE,7TH FLOOR,NEW YORK,NY 10016, USA. FU NCRR NIH HHS [RR 00168]; NIAID NIH HHS [AI 38573-02, AI 28147] NR 21 TC 347 Z9 352 U1 0 U2 14 PU NATURE PUBLISHING CO PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 SN 1078-8956 J9 NAT MED JI Nat. Med. PD OCT PY 1996 VL 2 IS 10 BP 1084 EP 1089 DI 10.1038/nm1096-1084 PG 6 WC Biochemistry & Molecular Biology; Cell Biology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Cell Biology; Research & Experimental Medicine GA VK995 UT WOS:A1996VK99500027 PM 8837605 ER PT J AU Parkinson, G Gunasekera, A Vojtechovsky, J Zhang, XP Kunkel, TA Berman, H Ebright, RH AF Parkinson, G Gunasekera, A Vojtechovsky, J Zhang, XP Kunkel, TA Berman, H Ebright, RH TI Aromatic hydrogen bond in sequence-specific protein DNA recognition SO NATURE STRUCTURAL BIOLOGY LA English DT Letter ID GENE ACTIVATOR PROTEIN; ESCHERICHIA-COLI; CAP; BENZENE; WATER; SITE; IDENTIFICATION; MUTATIONS; COMPLEX; BINDING AB An aromatic hydrogen bond-an interaction between the pi-electron cloud of an aromatic ring and a hydrogen-bond donor-can substitute for a conventional hydrogen bond in sequence-specific protein-DNA interactions and can contribute 0.5-1 kcal mol(-1) to binding and 1-2 kcal mol(-1) to specificity. C1 RUTGERS STATE UNIV, DEPT CHEM, NEW BRUNSWICK, NJ 08855 USA. RUTGERS STATE UNIV, WAKSMAN INST, NEW BRUNSWICK, NJ 08855 USA. NIEHS, NIH, RES TRIANGLE PK, NC 27709 USA. OI Ebright, Richard/0000-0001-8915-7140 NR 28 TC 64 Z9 68 U1 0 U2 5 PU NATURE PUBLISHING GROUP PI NEW YORK PA 75 VARICK ST, 9TH FLR, NEW YORK, NY 10013-1917 USA SN 1072-8368 J9 NAT STRUCT BIOL JI Nat. Struct. Biol. PD OCT PY 1996 VL 3 IS 10 BP 837 EP 841 DI 10.1038/nsb1096-837 PG 5 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA VK993 UT WOS:A1996VK99300006 PM 8836098 ER PT J AU Kawai, N McCarron, RM Spatz, M AF Kawai, N McCarron, RM Spatz, M TI Na+-K+-CL(-) cotransport system in brain capillary endothelial cells: Response to endothelin and hypoxia SO NEUROCHEMICAL RESEARCH LA English DT Article DE brain capillary endothelium; endothelin-1; hypoxia; Na+-K+-Cl- cotransport ID PROTEIN-KINASE-C; MIDDLE CEREBRAL-ARTERY; RAT-BRAIN; SIGNAL-TRANSDUCTION; DEPENDENT PATHWAY; SODIUM-TRANSPORT; ISCHEMIA; POTASSIUM; BARRIER; EDEMA AB Effect of endothelin-l and chemically induced hypoxia on Na+-K+-Cl- cotransport activity in cultured rat brain capillary endothelial cells was examined by using Rb-86(+) as a tracer for K+; bumetanide-sensitive K+ uptake was defined as Na+-K+-Cl- cotransport activity. Endothelin-l, phorbol 12-myristate 13-acetate (PMA), or thapsigargin increased Na+-K+-Cl- cotransport activity. A protein kinase C inhibitor, bisindolylmaleimide, inhibited PMA- and endothelin-1- (but not thapsigargin-) induced Na+-K+-Cl- cotransport activity, indicating the presence of both protein kinase C-dependent regulatory mechanisms and protein kinase C-independent mechanisms which involve intracellular Ca2+. Oligomycin, sodium azide, or antimycin A increased Na+-K+-Cl- cotransport activity by 80-200%. Oligomycin-induced Na+-K+-Cl- cotransport activity was reduced by an intracellular Ca2+ chelator (BAPTA/AM) but not affected by bisindolylmaleimide, suggesting the involvement of intracellular Ca2+, and not protein kinase C, in hypoxia-induced Na+-K+-Cl- cotransport activity. C1 NINCDS,NATL INST HLTH,STROKE BRANCH,BETHESDA,MD 20892. NR 30 TC 16 Z9 17 U1 0 U2 0 PU PLENUM PUBL CORP PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 SN 0364-3190 J9 NEUROCHEM RES JI Neurochem. Res. PD OCT PY 1996 VL 21 IS 10 BP 1259 EP 1266 DI 10.1007/BF02532403 PG 8 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA VN284 UT WOS:A1996VN28400016 PM 8923488 ER PT J AU Litvan, I Mangone, CA Werden, W Bueri, JA Estol, CJ Garcea, DO Rey, RC Sica, REP Hallett, M Bartko, JJ AF Litvan, I Mangone, CA Werden, W Bueri, JA Estol, CJ Garcea, DO Rey, RC Sica, REP Hallett, M Bartko, JJ TI Reliability of the NINDS Myotatic Reflex Scale SO NEUROLOGY LA English DT Article ID CATEGORICAL-DATA; AGREEMENT; ISSUES AB The assessment of deep tendon reflexes is useful for localization and diagnosis of neurologic disorders, but only a few studies have evaluated their reliability. We assessed the reliability of four neurologists, instructed in two different countries, in using the National Institute of Neurological Disorders and Stroke (NINDS) Myotatic Reflex Scale. To evaluate the role of training in using the scale, the neurologists randomly and blindly evaluated a total of 80 patients, 40 before and 40 after a training session. Inter- and intraobserver reliability were measured with kappa statistics. Our results showed substantial to near-perfect intraobserver reliability, and moderate-to-substantial interobserver reliability of the NINDS Myotatic Reflex Scale. The reproducibility was better for reflexes in the lower than in the upper extremities. Neither educational background nor the training session influenced the reliability of our results. The NINDS Myotatic Reflex Scale has sufficient reliability to be adopted as a universal scale. C1 HOSP RAMOS MEIJA, DIV NEUROL, BUENOS AIRES, DF, ARGENTINA. INST CARDIOVASC BUENOS AIRES, NEUROL SECT, BUENOS AIRES, DF, ARGENTINA. NINCDS, MED NEUROL BRANCH, NIH, BETHESDA, MD 20814 USA. NIMH, NIH, BETHESDA, MD 20892 USA. RP Litvan, I (reprint author), NINCDS, NEUROEPIDEMIOL BRANCH, NIH, FED BLDG, ROOM 714, BETHESDA, MD 20814 USA. OI Litvan, Irene/0000-0002-3485-3445 NR 22 TC 42 Z9 43 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 OCT PY 1996 VL 47 IS 4 BP 969 EP 972 PG 4 WC Clinical Neurology SC Neurosciences & Neurology GA VN295 UT WOS:A1996VN29500020 PM 8857728 ER PT J AU Sivakumar, K Dalakas, MC AF Sivakumar, K Dalakas, MC TI The spectrum of familial inclusion body myopathies in 13 families and a description of a quadriceps-sparing phenotype in non-Iranian Jews SO NEUROLOGY LA English DT Article ID DISTAL MYOPATHY; RIMMED VACUOLE; MYOSITIS; WELANDER AB The frequency, patterns of inheritance and clinical phenotypes of inherited myopathies with histologic features of rimmed vacuoles, tubulofilamentous inclusions and absence of inflammation (familial and hereditary inclusion body myopathy [f-IBM]) are poorly defined. Quadriceps sparing is a characteristic of f-IBM seen in the Iranian Jewish population. Among 101 patients with the feature of a red-rimmed vacuolar myopathy, characterized as inclusion body myopathy, seen during the last 4 years, we identified 13 families with f-IBM (12.8% frequency when one member per family was considered). Five families had an autosomal dominant and eight had an autosomal recessive form of inheritance. Among the latter group, five patients with early-onset disease (two Caucasian Americans, an Asian Indian, and two unrelated Iranian Jews) had the distinct feature of quadriceps sparing, which was confirmed by MRI of the thighs. Their disease began with weakness and atrophy of the foot extensors, forearm flexors, and first dorsal interossei muscles and progressed to the forearm flexors, girdle, and axial muscles, but spared the quadriceps. Serum CK was normal. Muscle biopsies showed rimmed vacuoles, small fibers in groups, amyloid deposition (in one patient), tubulofilaments, and no inflammation. Immunocytochemistry did not reveal abnormalities of various membrane or cytoskeletal proteins. Major histocompatibility complex (MHC) class I antigen was expressed only in a few degenerating fibers invaded by macrophages. T-cell infiltrates were not present. We conclude that in a large referral population, dominant and recessive hereditary and familial forms of IBM are not rare. Quadriceps-sparing myopathy appears to be a clinically distinct, autosomal recessive, nonimmune, distal vacuolar myopathy that is not limited to Iranian-Jewish ethnic groups. C1 NINCDS,NEUROMUSCULAR DIS SECT,NIH,BETHESDA,MD 20892. NR 22 TC 53 Z9 53 U1 0 U2 0 PU LITTLE BROWN CO PI BOSTON PA 34 BEACON STREET, BOSTON, MA 02108-1493 SN 0028-3878 J9 NEUROLOGY JI Neurology PD OCT PY 1996 VL 47 IS 4 BP 977 EP 984 PG 8 WC Clinical Neurology SC Neurosciences & Neurology GA VN295 UT WOS:A1996VN29500022 PM 8857730 ER PT J AU Rueckert, L Grafman, J AF Rueckert, L Grafman, J TI Sustained attention deficits in patients with right frontal lesions SO NEUROPSYCHOLOGIA LA English DT Article DE sustained attention; frontal lobe lesion ID HYPERACTIVITY DISORDER; WORKING-MEMORY; COMMISSUROTOMY; IMPAIRMENTS; BEHAVIOR; CAPACITY; CORTEX; SYSTEM; TASK AB Patients with frontal lobe lesions were compared to controls matched for age and education on several tests of sustained attention. One was a simple reaction time task requiring subjects to respond whenever they saw an 'X', one was a Continuous Performance Test that required subjects to respond to an 'X' but refrain from responding to other letters, and one involved reading a story and responding to a specified target. Patients with right frontal lesions showed longer RTs and missed more targets than control subjects for all three tests. In addition, right frontal patients got worse with time on the CPT. These results suggest a special role for the right frontal lobe in sustaining attention over time. Copyright (C) 1996 Elsevier Science Ltd C1 NINCDS, COGNIT NEUROSCI SECT, NATL INST HLTH, BETHESDA, MD 20892 USA. OI Grafman, Jordan H./0000-0001-8645-4457 NR 43 TC 158 Z9 159 U1 1 U2 6 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0028-3932 J9 NEUROPSYCHOLOGIA JI Neuropsychologia PD OCT PY 1996 VL 34 IS 10 BP 953 EP 963 DI 10.1016/0028-3932(96)00016-4 PG 11 WC Behavioral Sciences; Neurosciences; Psychology, Experimental SC Behavioral Sciences; Neurosciences & Neurology; Psychology GA VA417 UT WOS:A1996VA41700002 PM 8843061 ER PT J AU Prigatano, GP Weinstein, EA AF Prigatano, GP Weinstein, EA TI Edwin A. Weinstein's contributions to neuropsychological rehabilitation SO NEUROPSYCHOLOGICAL REHABILITATION LA English DT Review AB Disorders of self awareness following various brain injuries can greatly impede efforts at neuropsychological rehabilitation. Understanding and managing these disorders requires perspectives from neuropsychology, behavioural neurology and neuropsychiatry. This paper attempts to clarify Edwin A. Weinstein's pioneering work in this area and its impact on neuropsychological rehabilitation. C1 ST JOSEPHS HOSP,BARROW NEUROL INST,PHOENIX,AZ 85016. NIA,NEUROSCI LAB,BETHESDA,MD 20892. NR 35 TC 38 Z9 38 U1 0 U2 3 PU PSYCHOLOGY PRESS PI HOVE PA 27 CHURCH RD, HOVE, EAST SUSSEX, ENGLAND BN3 2FA SN 0960-2011 J9 NEUROPSYCHOL REHABIL JI Neuropsychol. Rehabil. PD OCT PY 1996 VL 6 IS 4 BP 305 EP 326 DI 10.1080/713755515 PG 22 WC Neurosciences; Psychology SC Neurosciences & Neurology; Psychology GA VT001 UT WOS:A1996VT00100005 ER PT J AU Gourovitch, ML Goldberg, TE Weinberger, DR AF Gourovitch, ML Goldberg, TE Weinberger, DR TI Verbal fluency deficits in patients with schizophrenia: Semantic fluency is differentially impaired as compared with phonologic fluency SO NEUROPSYCHOLOGY LA English DT Article ID MEMORY; PERFORMANCE; RETRIEVAL; DISORDERS; DISEASE; TESTS AB Patients with schizophrenia show deficits in phonologic (ability to name words that begin with a specific letter, e.g., F) and semantic (ability to name members of a category, e.g., ''animals'') fluency. Whereas the former deficit has been presumed to reflect a dysfunction of the frontal lobe, the latter has been linked to frontal and temporoparietal brain areas. These 2 verbal fluency measures were studied in a sample of 27 schizophrenia patients and 24 normal controls who were matched on age and a putative measure of premorbid intellectual ability. A 2-min production task of switching between letters and between categories measured demand for flexibility. On switching and nonswitching tasks controls produced more words during semantic versus phonologic fluency. Conversely, schizophrenia patients produced more words for letters than for categories, suggesting dysfunction of the frontal and temporoparietal areas of the brain. Furthermore, the greater impairment of semantic fluency may be related to a breakdown of semantic information processing beyond ''executive'' search and retrieval. C1 NIMH, CLIN BRAIN DISORDERS BRANCH, CTR NEUROSCI, INTRAMURAL RES PROGRAM, WASHINGTON, DC 20032 USA. NR 38 TC 63 Z9 63 U1 1 U2 1 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 0894-4105 EI 1931-1559 J9 NEUROPSYCHOLOGY JI Neuropsychology PD OCT PY 1996 VL 10 IS 4 BP 573 EP 577 PG 5 WC Psychology, Clinical; Neurosciences; Psychology SC Psychology; Neurosciences & Neurology GA VR665 UT WOS:A1996VR66500014 ER PT J AU Manji, HK Bersudsky, Y Chen, G Belmaker, RH Potter, WZ AF Manji, HK Bersudsky, Y Chen, G Belmaker, RH Potter, WZ TI Modulation of protein kinase C isozymes and substrates by lithium: The role of myo-inositol SO NEUROPSYCHOPHARMACOLOGY LA English DT Article DE lithium; manic-depressive; protein kinase C; G proteins; myo-inositol ID SIGNAL-TRANSDUCTION PATHWAYS; CEREBRAL-CORTEX SLICES; CDP-DIACYLGLYCEROL; RAT-BRAIN; PHOSPHATIDYLINOSITOL SYNTHESIS; PHOSPHOLIPID HYDROLYSIS; ADENYLATE-CYCLASE; DOWN-REGULATION; PHORBOL ESTER; SCIATIC-NERVE AB Lithium is the most effective treatment for reducing both the frequency and severity of recurrent affective episodes, but despite extensive research, the molecular mechanisms underlying its therapeutic actions have not been fully elucidated. Signal transduction pathways are in a pivotal postion in the central nervous system, able to affect the functional balance between neurotransmitter systems and have clearly been demonstrated to be targets of lithium's actions. We investigate the hypothesis that the action of chronic lithium on PKC isozymes and substrates may be secondary to its potent effect in inhibiting the recycling of inositol. Rats received lithium for 3 weeks and also myoinositol or saline twice daily via intracerebroventricular (ICV) injections. There was a significant interaction between chronic lithium and myo-inositol administration, with the chronic ICV administration of myo-inositol attenuating lithium's effects on PKC alpha, PKC epsilon, and on pertussin toxin-catalyzed [P-32]ADP-ribosylation. These results suggest that the effects of chronic lithium on signal transduction pathways may stem initially from its inhibition of inositol-1-phosphatase. Given the critical role of PKC isozymes and G proteins in modulating intracellular cross-talk between neurotransmitters systems and thereby the integrative functions of the CNS, future studies using other inhibitors of inositol monophosphatases are warranted, and offer the hope for the development of more potent and more rapidly acting mood-stabilizing drugs. C1 BEERSHEVA MENTAL HLTH CTR,BEER SHEVA,ISRAEL. NIMH,CLIN PHARMACOL SECT,BETHESDA,MD 20892. RP Manji, HK (reprint author), WAYNE STATE UNIV,SCH MED,DEPT PSYCHIAT & BEHAV NEUROSCI,DETROIT,MI 48201, USA. RI Chen, Guang/A-2570-2017 NR 64 TC 68 Z9 70 U1 1 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 SN 0893-133X J9 NEUROPSYCHOPHARMACOL JI Neuropsychopharmacology PD OCT PY 1996 VL 15 IS 4 BP 370 EP 381 DI 10.1016/0893-133X(95)00243-7 PG 12 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA VK316 UT WOS:A1996VK31600007 PM 8887991 ER PT J AU Bergqvist, PBF Heyes, MP Apelqvist, G Butterworth, RF Bengtsson, F AF Bergqvist, PBF Heyes, MP Apelqvist, G Butterworth, RF Bengtsson, F TI Brain extracellular quinolinic acid in chronic experimental hepatic encephalopathy as assessed by in vivo microdialysis: Acute effects of L-tryptophan SO NEUROPSYCHOPHARMACOLOGY LA English DT Article DE hepatic encephalopathy; in vivo cerebral microdialysis; kynurenines; liver disease; portacaval anastomosis; quinolinic acid ID CHRONIC PORTACAVAL ANASTOMOSIS; FREELY-MOVING RATS; CHAIN AMINO-ACIDS; CEREBROSPINAL-FLUID; INVIVO MICRODIALYSIS; SEROTONIN RELEASE; PLASMA; METABOLISM; BLOOD; QUANTIFICATION AB Increased brain quinolinic acid (QUIN) levels have been suggested to play a role in hepatic encephalopathy (HE). Previous brain tissue studies have been unable to confirm this hypothesis. Because QUIN is a potent NMDA-receptor agonist, it also is relevant to determine brain extracellular QUIN levels in HE. For this purpose, we assessed frontal neocortical extracellular QUIN levels by in vivo microdialysis in rats subjected to a portacaval shunt (PCS). We also evaluated the acute effects of altered L-tryptophan (L-TRP) availability on brain extracellular QUIN levels. The basal extracellular L-TRP levels were significantly (p < .001) higher in the PCS rats than in the sham-operated controls. However, the QUIN level (p < .05) and the QUIN to L-TRP ratio (p < .01) were significantly lower in the PCS rats. Elevated L-TRP availability increased the QUIN levels to a similar degree in both sham and PCS rats. This study, in conjunction with our previous results, does thereby not support a major involvement of QUIN in the pathogenesis of HE. C1 LUND UNIV,DEPT CLIN PHARMACOL,LUND,SWEDEN. NIMH,CLIN SCI LAB,ANALYT BIOCHEM SECT,BETHESDA,MD 20892. UNIV MONTREAL,ST LUC HOSP,NEUROSCI RES UNIT,MONTREAL,PQ,CANADA. NR 39 TC 13 Z9 13 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 SN 0893-133X J9 NEUROPSYCHOPHARMACOL JI Neuropsychopharmacology PD OCT PY 1996 VL 15 IS 4 BP 382 EP 389 DI 10.1016/0893-133X(95)00256-D PG 8 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA VK316 UT WOS:A1996VK31600008 PM 8887992 ER PT J AU Kuo, H Ingram, DK Walker, LC Tian, M Hengemihle, JM Jucker, M AF Kuo, H Ingram, DK Walker, LC Tian, M Hengemihle, JM Jucker, M TI Similarities in the age-related hippocampal deposition of periodic acid-Schiff-positive granules in the senescence-accelerated mouse P8 and C57BL/6 mouse strains SO NEUROSCIENCE LA English DT Article DE hippocampus; heparan sulfate proteoglycan; astrocytes; neurodegeneration; aging; Alzheimer's disease ID HEPARAN-SULFATE PROTEOGLYCAN; ALZHEIMERS-DISEASE; WALLERIAN DEGENERATION; FIBRILLAR MATERIAL; BRAIN ATROPHY; MODEL; MICE; ASTROCYTES; SAM-P/8; RAT AB With advancing age, clusters of abnormal granules positive for periodic acid-Schiff appear in the hippocampus bf C57BL/6 (B6) mice and the senescence-accelerated mouse (SAM) P8. The granules can also be visualized with a polyclonal antibody to a 110,000 mol. wt laminin-binding protein and stain specifically with a monoclonal antibody to heparan sulfate proteoglycan. The present study used light- and electron-microscopic analysis to compare the staining and morphological properties of these granules in SAM P8 hippocampus with those in B6 hippocampus at different ages. The results of the light-microscopic analysis revealed that granules in SAM P8 and B6 had similar morphology, staining characteristics and distribution patterns, and appeared to have a close association with astrocytic processes. The onset of granules in SAM P8 mice (at two to three months of age) was earlier than that observed in B6 mice (al four to six months of age), but the maximum incidence was similar in both strains. Electron-microscopic analysis revealed that the granules in SAM P8 and B6 mice also had a very similar ultrastructure. Granules in both strains were surrounded by a discontinuous membrane and contained mostly crystalline-like, degenerated material. The successive ultrastructural changes from the exterior to interior of the granules suggest that the degenerative process was initiated outside the granules and that degenerative structures migrate inward. Astrocytes and heparan sulfate proteoglycan are closely associated with beta-amyloid deposits in Alzheimer's disease. The presence of astrocyte-associated heparan sulfate proteoglycan-positive material in aged SAM P8 and B6 mice might model age-related alterations in glia function possibly involved in human cerebral amyloidogenesis. C1 NIA, NIH, CTR GERONTOL RES, NATHAN W SHOCK LABS, BALTIMORE, MD 21224 USA. JOHNS HOPKINS UNIV, SCH MED, DEPT PATHOL, NEUROPATHOL LAB, BALTIMORE, MD 21205 USA. UNIV GREIFSWALD, DEPT NEUROL, GREIFSWALD, GERMANY. RI Walker, L/J-6541-2015 OI Walker, L/0000-0001-9166-3261 FU NIA NIH HHS [AG 05146]; NINDS NIH HHS [NS 20471] NR 27 TC 22 Z9 28 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0306-4522 J9 NEUROSCIENCE JI Neuroscience PD OCT PY 1996 VL 74 IS 3 BP 733 EP 740 DI 10.1016/0306-4522(96)00169-8 PG 8 WC Neurosciences SC Neurosciences & Neurology GA VH282 UT WOS:A1996VH28200009 PM 8884769 ER PT J AU Drago, J Gerfen, CR Westphal, H Steiner, H AF Drago, J Gerfen, CR Westphal, H Steiner, H TI D1 dopamine receptor-deficient mouse: Cocaine-induced regulation of immediate-early gene and substance P expression in the striatum SO NEUROSCIENCE LA English DT Article DE c-fos; zif 268; enkephalin; dynorphin; neuropeptide; amphetamine ID MESSENGER-RNA EXPRESSION; TRANSCRIPTION FACTOR GENES; C-FOS INDUCTION; RAT STRIATUM; BEHAVIORAL-RESPONSES; STRIATOPALLIDAL NEURONS; MATRIX COMPARTMENTS; NUCLEUS-ACCUMBENS; MUTANT MICE; DYNORPHIN AB Psychomotor stimulants such as cocaine alter gene expression in neurons of the striatum. Whereas many of these effects are mediated by D1 dopamine receptors, the involvement of other dopamine receptor subtypes or neurotransmitters is likely. To distinguish between these possibilities, regulation by cocaine of immediate-early genes and genes encoding neuropeptides was analysed in mice that lack functional Di receptors. Gene expression was examined with in situ hybridization histochemistry. In these animals; cocaine failed to induce the immediate-early genes c-Sos and zif 268. In contrast, substance P expression was abnormally increased by this drug. These results demonstrate that some of the effects of cocaine on gene regulation are mediated via D1 receptor-dependent mechanisms, as evidenced by the absence of immediate-early gene induction in D1-deficient mice, whereas others also involve additional, non-D1 receptor mechanisms, as shown for substance P expression. C1 NIMH,NEUROPHYSIOL LAB,BETHESDA,MD 20892. NICHHD,LAB MAMMALIAN GENES & DEV,BETHESDA,MD 20892. NR 62 TC 111 Z9 113 U1 0 U2 6 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, ENGLAND OX5 1GB SN 0306-4522 J9 NEUROSCIENCE JI Neuroscience PD OCT PY 1996 VL 74 IS 3 BP 813 EP 823 DI 10.1016/0306-4522(96)00145-5 PG 11 WC Neurosciences SC Neurosciences & Neurology GA VH282 UT WOS:A1996VH28200017 PM 8884777 ER PT J AU Boulay, SF Sood, VK Rayeq, MR Zeeberg, BR Eckelman, WC AF Boulay, SF Sood, VK Rayeq, MR Zeeberg, BR Eckelman, WC TI Autoradiographic evidence that (R)-3-quinuclidinyl (S)-4-fluoromethylbenzilate ((R,S)-FMeQNB) displays in vivo selectivity for the muscarinic m2 subtype SO NUCLEAR MEDICINE AND BIOLOGY LA English DT Article DE pharmacokinetic modeling; emission tomographic neuroreceptor quantitation; radioiodinated (R,S)4IQNB; (R,S)-FMeQNB ID RAT-BRAIN; ALZHEIMERS-DISEASE; ACETYLCHOLINE-RECEPTOR; INVIVO; BINDING; COMPETITION; TOMOGRAPHY; BENZILATE; PROTEINS; INVITRO AB Alzheimer's disease (AD) involves selective loss of muscarinic m2, but not m1, subtype neuroreceptors in cortical and hippocampal regions of the human brain. Until recently, emission tomographic study of the loss of m2 receptors in AD has been limited by the absence of available m2 selective radioligands that can penetrate the blood brain barrier. We now demonstrate the in vivo m2 selectivity of a fluorinated derivative of QNB, (R)-3-quinuclidinyl (S)-4-fluoromethylbenzilate ((R,S)-FMeQNB), by studying autoradiographically the in vivo inhibition of radioiodinated (R)-3-quinuclidinyl (S)-4-iodobenzilate ((R,S)-[I-125]IQNB) binding by unlabelled (R,S)-FMeQNB. In the absence of (R,S)-FMeQNB, (R,S)-[I-125]IQNB labels brain regions in proportion to the total muscarinic receptor concentration; in the presence of 75 nmol of (R,S)-FMeQNB, (R,S)-[I-125]IQNB labelling in those brain regions containing predominantly m2 subtype is reduced to background levels. We conclude that (R,S)-FMeQNB is m2 selective in vivo, and that (R,S)-[F-18]FMeQNB may be of potential use in positron emission tomographic (PET) study of the loss of m2 receptors in AD. Copyright (C) Elsevier Science Inc. C1 GEORGE WASHINGTON UNIV,MED CTR,DEPT RADIOL,SECT RADIOPHARMACEUT CHEM,WASHINGTON,DC 20037. NIH,PET DEPT,CTR CLIN,BETHESDA,MD 20892. RI Sood, Vinay/B-7109-2008 FU NINDS NIH HHS [NS22215] NR 41 TC 9 Z9 9 U1 1 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, ENGLAND OX5 1GB SN 0883-2897 J9 NUCL MED BIOL JI Nucl. Med. Biol. PD OCT PY 1996 VL 23 IS 7 BP 889 EP 896 DI 10.1016/S0969-8051(96)00121-7 PG 8 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA VT930 UT WOS:A1996VT93000005 PM 8971856 ER PT J AU Hickey, CA Cliver, SP McNeal, SF Hoffman, HJ Goldenberg, RL AF Hickey, CA Cliver, SP McNeal, SF Hoffman, HJ Goldenberg, RL TI Prenatal weight gain patterns and birth weight among nonobese black and white women SO OBSTETRICS AND GYNECOLOGY LA English DT Article ID FETAL GROWTH-RETARDATION; INTRAUTERINE GROWTH; PRETERM DELIVERY; PREGNANCY; TERM; AGE AB Objective: To examine the association between prenatal weight gain patterns and birth weight using Institute of Medicine (IOM) guidelines. Methods: Data from a prospective follow-up study of risk factors for fetal growth restriction were used to examine the impact of low weight gain on mean birth weight. A total of 415 nonobese (body mass index [BMI] less than 26) black (n = 275) and white (n = 140) women who delivered at term were included in this analysis. Linear regression analysis was used to examine the impact of low first-trimester gain (less than 2.3 kg with low BMI [less than 19.8]; less than 1.6 kg with normal BMI [19.8-26.0]) and low second- and third-trimester rates of gain (less than 0.38 kg/week with low BMI; less than 0.37 kg/week with normal BMI) on mean birth weight while controlling for selected sociodemographic and reproductive variables. Results: Patterns with low gain in the first and second or in the second and third trimesters were associated with significant decreases in mean birth weight, ranging from 206 to 265 g; low gain in only the first or third trimester was not associated with a significant decrease in mean in birth weight. The impact of low gain on mean birth weight varied by ethnic group. Conclusion: These observations suggest that inadequate patterns of prenatal weight gain, defined by IOM guidelines, are associated with decreased birth weight, particularly when the patterns involve low second-trimester gain. C1 UNIV ALABAMA,SCH MED,DEPT OBSTET & GYNECOL,CTR OBSTET RES,BIRMINGHAM,AL. NIDOCD,EPIDEMIOL STAT & DATA SYST BRANCH,NIH,BETHESDA,MD. RP Hickey, CA (reprint author), UNIV ALABAMA,SCH PUBL HLTH,DEPT MATERNAL & CHILD HLTH,112 MORTIMER JORDAN HALL,1825 UNIV BLVD,BIRMINGHAM,AL 35294, USA. FU NICHD NIH HHS [1-HD-4-2811]; PHS HHS [290-92-0055, MCJ-9040] NR 20 TC 57 Z9 57 U1 0 U2 4 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 SN 0029-7844 J9 OBSTET GYNECOL JI Obstet. Gynecol. PD OCT PY 1996 VL 88 IS 4 BP 490 EP 496 DI 10.1016/0029-7844(96)00262-1 PN 1 PG 7 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA VK449 UT WOS:A1996VK44900002 PM 8841205 ER PT J AU Ananth, CV Wilcox, AJ Savitz, DA Bowes, WA Luther, ER AF Ananth, CV Wilcox, AJ Savitz, DA Bowes, WA Luther, ER TI Effect of maternal age and parity on the risk of uteroplacental bleeding disorders in pregnancy SO OBSTETRICS AND GYNECOLOGY LA English DT Article ID ABRUPTIO PLACENTAE; PREVIA AB Objective: To examine the risk of placental abruption, placenta previa, and uterine bleeding of unknown etiology in relation to advanced maternal age and parity in a large, population-based study. Methods: Data for this study were derived from the Nova Scotia Atlee perinatal provincial data base, Canada, an ongoing project on human reproduction. Women who delivered between 1980 and 1993 (n = 123,941) in the province of Nova Scotia were included in the study, with the exception of pregnancies resulting in multiple births (12 = 2859) and those missing data on maternal age or parity (n = 14). Multivariable logistic regression models based on the method of generalized estimating equations were used to generate odds ratios after adjustment for multiple confounders. Results: The frequency of abruption was increased slightly among younger women (relative risk [RR] 1.3, 95% confidence interval [CI] 1.0-1.7), compared with women ages 25-29 years, but there was no increase with advancing maternal age. In contrast, the risk of placenta previa increased dramatically with advancing maternal age, with women older than 40 years having a nearly ninefold greater risk than women under the age of 20, after adjustment for potential confounders, including parity. Uterine bleeding of unknown etiology was not associated with advanced maternal age, except for a slight increase among women over 40 (RR 1.3, 95% CI 1.0-1.6). The risk of placenta previa and placental abruption was increased with higher parity among younger women only, but uterine bleeding of unknown etiology was more weakly associated with higher parity. In addition, an analysis of the joint effects of age and parity on placental abruption indicated a strong parity effect for women under 30 years, whereas the risk of placenta previa increased with increasing parity up to age 35 years. Uterine bleeding of unknown etiology also indicated a parity effect that was restricted to women under 25 years. Conclusion: Multiparity is associated with the risk of placenta previa and, to a lesser extent, placental abruption, but not with other uterine bleeding. Increasing maternal age is associated independently with the risk of placenta previa, but not with either of the other two conditions. Finally, the increased risks of uteroplacental bleeding disorders with advanced parity among the younger women (ie, 20-25 years, parity 3+) may reflect effects of close pregnancy spacing, or confounding by unmeasured factors that characterize women who have many pregnancies at a relatively young age. Overall, the findings suggest that the three uteroplacental bleeding disorders do not share a common etiology in relation to maternal age and parity, and that placenta previa is linked to aging of the uterus and the effects of repeated pregnancies. C1 UNIV N CAROLINA,SCH PUBL HLTH,DEPT EPIDEMIOL,CHAPEL HILL,NC. UNIV N CAROLINA,CAROLINA POPULAT CTR,CHAPEL HILL,NC. NIEHS,EPIDEMIOL BRANCH,RES TRIANGLE PK,NC 27709. UNIV N CAROLINA,DEPT OBSTET & GYNECOL,DIV MATERNAL & FETAL MED,CHAPEL HILL,NC. DALHOUSIE UNIV,DEPT OBSTET & GYNECOL,DIV MATERNAL & FETAL MED,HALIFAX,NS,CANADA. OI Wilcox, Allen/0000-0002-3376-1311 NR 26 TC 68 Z9 70 U1 1 U2 3 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 SN 0029-7844 J9 OBSTET GYNECOL JI Obstet. Gynecol. PD OCT PY 1996 VL 88 IS 4 BP 511 EP 516 DI 10.1016/0029-7844(96)00236-0 PN 1 PG 6 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA VK449 UT WOS:A1996VK44900005 PM 8841208 ER PT J AU Sherer, DM Fromberg, RA Divon, MY AF Sherer, DM Fromberg, RA Divon, MY TI Prenatal ultrasonographic assessment of the ductus venosus: A review SO OBSTETRICS AND GYNECOLOGY LA English DT Review ID VELOCITY WAVE-FORMS; BLOOD-FLOW VELOCITY; INFERIOR VENA-CAVA; FETAL CIRCULATION; DOPPLER ASSESSMENT; HUMAN FETUS; ARTERIAL; VELOCIMETRY; TRANSFUSION AB Objective: To review current data pertaining to prenatal ultrasonography of the ductus venosus. Data Sources: We identified English-language studies regarding prenatal ultrasonography and the fetal ductus venosus. The studies were obtained from a MEDLINE search for the period 1966 through March 1996. Additional sources were identified through cross-referencing. Methods of Study Selection: We reviewed all published reports, case studies, and articles regarding ultrasonographic morphology, physiology, and pathophysiology of the fetal ductus venosus. Tabulation, Integration, and Results: Knowledge of the function of the fetal ductus venosus in both normal and abnormal fetal conditions is increased by prenatal ultrasonographic data. Altered ductus venosus hemodynamics may be noted in various medical conditions that include fetal growth restriction, twin-twin transfusion, invasive diagnostic procedures (chorionic villus sampling and fetal blood sampling), fetal anemia and transfusion, complex cardiac structural anomalies, and cardiac arrhythmias. Conclusion: blood flow velocity in the fetal ductus venosus reflects the pressure gradient between the umbilical vein and the cardiac atria. Knowledge of pathophysiologic hemodynamic changes of blood now in this vessel obtainable by prenatal ultrasonography in conjunction with fetal disease may assist diagnosis and management. C1 NICHHD,NIH,PERINATOL RES FACIL,INTRAMURAL DIV,DIV MATERNAL FETAL MED,WASHINGTON,DC. RP Sherer, DM (reprint author), ALBERT EINSTEIN COLL MED,MONTEFIORE MED CTR,DIV MATERNAL FETAL MED,DEPT OBSTET & GYNECOL,BRONX,NY 10461, USA. NR 32 TC 9 Z9 12 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 SN 0029-7844 J9 OBSTET GYNECOL JI Obstet. Gynecol. PD OCT PY 1996 VL 88 IS 4 BP 626 EP 632 DI 10.1016/0029-7844(96)00181-0 PN 1 PG 7 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA VK449 UT WOS:A1996VK44900029 PM 8841232 ER PT J AU Wiener, LS Battles, HB Heilman, N Sigelman, CK Pizzo, PA AF Wiener, LS Battles, HB Heilman, N Sigelman, CK Pizzo, PA TI Factors associated with disclosure of diagnosis to children with HIV/AIDS SO PEDIATRIC AIDS AND HIV INFECTION-FETUS TO ADOLESCENT LA English DT Article ID ACQUIRED-IMMUNODEFICIENCY-SYNDROME; HIV-INFECTION; AIDS; CANCER; DEATH; COMMUNICATION; ADOLESCENTS; PHYSICIANS; KNOWLEDGE; ATTITUDES AB Disclosure of the diagnosis of human immunodeficiency virus (HIV) infection or acquired immunodeficiency syndrome (AIDS) to a child is a controversial and emotionally laden issue. To understand the factors that affect the process of disclosure and its consequences, we studied 99 parent-child dyads recruited from patients being treated at the National Cancer Institute (NCI). Parents and HIV-infected children were interviewed and administered several standardized measures. Parental depression, family environment, social support satisfaction, socioeconomic status, child and parent gender, child's age, parental HIV serostatus, and disease severity were used to predict disclosure status. Results indicate that the majority of caregivers do disclose the diagnosis to the child, usually with no ill effects, and that age is the most significant predictor of whether or not a child has been told. The Centers for Disease Control and Prevention currently estimate that there are over 6611 children with AIDS (under age 13), and 2184 adolescents with AIDS (ages 13-19) in America.(1) As an increasing number of children who are born infected with HIV live to older ages, the question of when and how to talk with them about their illness becomes more crucial.(2,3) In addition to the growing number of children infected with HIV, there are many thousands of children profoundly affected by the impact of this disease on a close family member-a mother, father, sibling, or other relative in the kinship network.(4) Yet, the initial reaction most adults have upon learning of their own, or of a family member's, HIV diagnosis is that the diagnosis must be kept a closely guarded secret. One reason frequently cited by parents and family members is their fear that the stigma of AIDS will have a negative impact on their children arid their families. Disclosure of an HIV diagnosis to a child is a controversial and emotionally laden issue in the pediatric health-care community as well. However, no systematic research has studied the issues that surround disclosure of an HIV diagnosis to the patient and the factors that predict disclosure. RP Wiener, LS (reprint author), NCI,PEDIAT BRANCH,BLDG 10,BETHESDA,MD 20892, USA. NR 49 TC 52 Z9 52 U1 2 U2 10 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 SN 1045-5418 J9 PEDIATR AIDS HIV INF JI Pediatr. AIDS HIV Infect.-Fetus Adolesc. PD OCT PY 1996 VL 7 IS 5 BP 310 EP 324 PG 15 WC Immunology; Infectious Diseases; Pediatrics SC Immunology; Infectious Diseases; Pediatrics GA VP368 UT WOS:A1996VP36800002 PM 11361489 ER PT J AU Falkner, B Daniels, SR Loggie, JMH Horan, MJ Prineas, RJ Rosner, B Sinaiko, AR Roccella, EJ Anderson, DE AF Falkner, B Daniels, SR Loggie, JMH Horan, MJ Prineas, RJ Rosner, B Sinaiko, AR Roccella, EJ Anderson, DE TI Update on the 1987 Task Force Report on High Blood Pressure in Children and Adolescents: A Working Group Report from the National High Blood Pressure Education Program SO PEDIATRICS LA English DT Article DE hypertension; children; adolescents; treatment; normative blood pressure tables ID BOGALUSA HEART; OBESE CHILDREN; WEIGHT-LOSS; FOLLOW-UP; FAMILIAL AGGREGATION; CARDIAC INVOLVEMENT; HYPERTENSION; CHILDHOOD; SODIUM; ANGIOTENSIN AB Background. The ''Report of the Second Task Force on Blood Pressure Control in Children-1987'' developed normative blood pressure (BP) data for children and adolescents. These normative data are used to classify BP levels. Since 1987, additional BP data in children and adolescents, the use of newer classes of drugs, and the role of primary prevention of hypertension have expanded the body of knowledge regarding the classification and treatment of hypertension in the young. Objective. To report new normative BP data in children and adolescents and to provide additional information regarding the diagnosis, treatment, and prevention of hypertension in children. Methods. A working group was appointed by the director of the National Heart, Lung, and Blood Institute as chair of the National High Blood Pressure Education Program (NHBPEP) Coordinating Committee. Data on children from the 1988 through 1991 National Health and Nutrition Examination Survey III and nine additional national data sets were combined to develop normative BP tables. The working group members produced initial draft documents that were reviewed by NHBPEP Coordinating Committee representatives as well as experts in pediatrics, cardiology, and hypertension. This reiterative process occurred for 12 draft documents. The NHBPEP Coordinating Committee discussed the report, and additional comments were received. Differences of opinion were adjudicated by the chair of the working group. The final report was sent to representatives of the 44 organizations on the NHBPEP Coordinating Committee for vote. It was approved unanimously by the NHBPEP Coordinating Committee on October 2, 1995. Conclusions. This report provides new normative BP tables for children and adolescents, which now include height percentiles, age, and gender. The fifth Korotkoff sound is now used to define diastolic BP in children and adolescents. New charts have been developed to guide practicing clinicians in antihypertensive drug therapy selection. The primary prevention of hypertension in these age groups is discussed. A statement on public health considerations in the treatment of children and adolescents is provided. C1 NHLBI, NATL HIGH BLOOD PRESSURE EDUC PROGRAM, BETHESDA, MD 20892 USA. MED COLL PENN & HAHNEMANN UNIV, PHILADELPHIA, PA USA. UNIV CINCINNATI, CHILDRENS HOSP & MED CTR, CINCINNATI, OH 45221 USA. NHLBI, DIV HEART & VASC DIS, BETHESDA, MD 20892 USA. UNIV MIAMI, SCH MED, CHAIR EPIDEMIOL & PUBL HLTH, MIAMI, FL USA. HARVARD UNIV, SCH MED, CHANNING LAB, BOSTON, MA 02115 USA. UNIV MINNESOTA, SCH MED, DEPT PEDIAT, MINNEAPOLIS, MN 55455 USA. ROW SCI INC, NATL HIGH BLOOD PRESSURE EDUC PROGRAM, ROCKVILLE, MD USA. NR 70 TC 793 Z9 857 U1 3 U2 27 PU AMER ACAD PEDIATRICS PI ELK GROVE VILLAGE PA 141 NORTH-WEST POINT BLVD,, ELK GROVE VILLAGE, IL 60007-1098 USA SN 0031-4005 J9 PEDIATRICS JI Pediatrics PD OCT PY 1996 VL 98 IS 4 BP 649 EP 658 PG 10 WC Pediatrics SC Pediatrics GA VN090 UT WOS:A1996VN09000001 ER PT J AU Long, RM AF Long, RM TI News from the National Institute of General Medical Sciences (NIGMS) SO PHARMACEUTICAL RESEARCH LA English DT News Item RP Long, RM (reprint author), NIGMS,NIH,DIV PHARMACOL PHYSIOL & BIOL CHEM,PHARMACOL & PHYSIOL SCI BRANCH,BETHESDA,MD 20892, USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU PLENUM PUBL CORP PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 SN 0724-8741 J9 PHARMACEUT RES JI Pharm. Res. PD OCT PY 1996 VL 13 IS 10 BP 1425 EP 1426 PG 2 WC Chemistry, Multidisciplinary; Pharmacology & Pharmacy SC Chemistry; Pharmacology & Pharmacy GA VN257 UT WOS:A1996VN25700001 ER PT J AU Venkatesh, S Li, JM Xu, YH Vishnuvajjala, R Anderson, BD AF Venkatesh, S Li, JM Xu, YH Vishnuvajjala, R Anderson, BD TI Intrinsic solubility estimation and pH-solubility behavior of cosalane (NSC 658586), an extremely hydrophobic diprotic acid SO PHARMACEUTICAL RESEARCH LA English DT Article DE cosalane; solubility; facilitated dissolution; partition coefficient; group contribution approach; pH-solubility behavior ID ANTI-HIV AGENT; INHIBITION; PROTEASE; DESIGN AB Purpose. The selection of cosalane (NSC 658586) by the National Cancer Institute for further development as a potential drug candidate for the treatment of AIDS led to the exploration of the solubility behavior of this extremely hydrophobic drug, which has an intrinsic solubility (S-0) approaching 1 ng/ml. This study describes attempts to reliably measure the intrinsic solubility of cosalane and examine its pH-solubility behavior. Methods. S-0 was estimated by 5 different strategies: (a) direct determination in an aqueous suspension; (b) facilitated dissolution; (c) estimation from the octanol/water partition coefficient and octanol solubility; (d) application of an empirical equation based on melting point and partition coefficient; and (e) estimation from the hydrocarbon solubility and functional group contributions for transfer from hydrocarbon to water. Results. S-0 estimates using these five methods varied over a 5 x 10(9)-fold range. Method (a) yielded the highest values, two-orders of magnitude greater than those obtained by method (b) (facilitated dissolution, 1.4 +/- 0.5 ng/ml). Method (c) gave a value 20-fold higher while that from method (d) was in fair agreement with that from facilitated dissolution. Method (e) yielded a value several orders-of-magnitude lower than other methods. A molecular dynamics simulation suggests that folded conformations not accounted for by group contributions may reduce cosalane's effective hydrophobicity. Ionic equilibria calculations for this weak diprotic acid suggested a 100-fold increase in solubility per pH unit increase. The pH-solubility profile of cosalane at 25 degrees C agreed closely with theory. Conclusions. These studies highlight the difficulty in determining solubility of very poorly soluble compounds and the possible advantage of the facilitated dissolution method. The diprotic nature of cosalane enabled a solubility enhancement of > 10(7)-fold by simple pH adjustment. C1 UNIV UTAH,DEPT PHARMACEUT & PHARMACEUT CHEM,SALT LAKE CITY,UT 84112. NCI,NIH,BETHESDA,MD 20892. FU NCI NIH HHS [N0I-CM-27720] NR 18 TC 16 Z9 16 U1 0 U2 2 PU PLENUM PUBL CORP PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 SN 0724-8741 J9 PHARMACEUT RES JI Pharm. Res. PD OCT PY 1996 VL 13 IS 10 BP 1453 EP 1459 DI 10.1023/A:1016059008464 PG 7 WC Chemistry, Multidisciplinary; Pharmacology & Pharmacy SC Chemistry; Pharmacology & Pharmacy GA VN257 UT WOS:A1996VN25700005 PM 8899834 ER PT J AU Chen, HT Ji, ZZ Wong, LK Siuda, JF Narayanan, VL AF Chen, HT Ji, ZZ Wong, LK Siuda, JF Narayanan, VL TI Synthesis, antiinflammatory, and cytotoxic activities of 2-alkyl and 2-benzyl-2-dimethylaminomethyl-5(E)-arylidene cyclopentanone hydrochlorides SO PHARMACEUTICAL RESEARCH LA English DT Article DE 2-aminomethyl-5-(E)-arylidene cyclopentanones; Mannich bases; antiinflammatory; analgesic; cytotoxic ID PHENYL-3-METHOXY-4-HYDROXY STYRYL KETONES; METHYL-3-METHOXY-4-HYDROXY AB Purpose. A series of 2-substituted-2-dimethylaminomethyl-5-(E)-arylidene cyclopentanones, 4 were synthesized. The main objective of this investigation was to explore the structural parameters necessary for antiinflammatory activity in this series of compounds, while keeping cytotoxic action to a minimum. Methods. The target compounds were synthesized in two steps commencing with 2-alkyl-cyclopentanones. Antiinflammatory, analgesic and cytotoxic activities were determined in rats. Cytotoxic results were examined in human cell lines. Results. Eight of the eighteen synthetic substances possessed significant antiinflammatory activity and twelve showed appreciable analgesic action. Cytotoxicity was minimal or nonexistent for most of the compounds. The stability of one of the compounds, 4b in both aqueous and non-aqueous media, and an amine exchange reaction with aniline wee used to explain the observed antiinflammatory and cytotoxic activities. Conclusions. Unlike monosubstituted aminomethyl groups (Mannich bases) at the 2-position of 5-arylidene-2-cyclopentanones, a second substituent at the 2-position increases stability of the Mannich base and significantly decreases cytotoxic activity, Antiinflammatory and analgesic action is retained in many of the compounds, thus strongly indicating that these desired pharmacological results can be obtained without untoward damage to cells. C1 UNIV PITTSBURGH,SCH PHARM,PITTSBURGH,PA 15261. SHENYANG COLL PHARM,SHENYANG,PEOPLES R CHINA. NCI,DRUG SYNTHESIS & CHEM BRANCH,NIH,BETHESDA,MD 20205. NR 17 TC 9 Z9 12 U1 0 U2 0 PU PLENUM PUBL CORP PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 SN 0724-8741 J9 PHARMACEUT RES JI Pharm. Res. PD OCT PY 1996 VL 13 IS 10 BP 1482 EP 1487 DI 10.1023/A:1016067210281 PG 6 WC Chemistry, Multidisciplinary; Pharmacology & Pharmacy SC Chemistry; Pharmacology & Pharmacy GA VN257 UT WOS:A1996VN25700009 PM 8899838 ER PT J AU Yokoi, T Kosaka, Y Chida, M Chiba, K Nakamura, H Ishizaki, T Kinoshita, M Sato, K Gonzalez, FJ Kamataki, T AF Yokoi, T Kosaka, Y Chida, M Chiba, K Nakamura, H Ishizaki, T Kinoshita, M Sato, K Gonzalez, FJ Kamataki, T TI A new CYP2D6 allele with a nine base insertion in exon 9 in a Japanese population associated with poor metabolizer phenotype SO PHARMACOGENETICS LA English DT Article DE Xb alpha I RFLP; yeast expression system; population study; bufuralol 1'-hydroxylase; magnetic polymorphism ID GENETIC-POLYMORPHISM; DEBRISOQUINE OXIDATION; SPARTEINE METABOLISM; MUTANT ALLELES; IDENTIFICATION; METOPROLOL; EXPRESSION; CYTOCHROME-P450; FRAGMENT; CHINESE AB The CYP2D6 gene of a Japanese sparteine poor metabolizer (PM, proband) showing a urinary metabolic ratio of 31.6 was analysed, and a heterozygous CYP2D6(D), a deletional was found by restriction fragment length polymorphism analysis with Xba I enzyme, PM did not have any other previously described mutations in the CYP2D6 gene causing the loss of catalytic activity of the CYP2D6 enzyme. Thus, a possible new allele(s) responsible the PM phenotype was analysed. The results indicated that the PM possessed a new 9-base in exon 9, designated CYP2D6(J9). The CYP2D6(J9) and CPP2D6(D) alleles were to be inherited from the mother [2D6(W)/2D6(J9)] and the father [2D6(W)/2D6(D)], respectively. The 9-base insertion caused a large increase in the apparent K-m value for bufuralol 1'-hydroxylation as examined by expression of the enzyme protein in yeast, Four of 300 Japanese carried a heterozygous CYP2D6(J9) allele (0.7%, 4/600 chromosomes) as determined a polymerase chain reaction analysis. C1 HOKKAIDO UNIV,FAC PHARMACEUT SCI,DIV DRUG METAB,KITA KU,SAPPORO,HOKKAIDO 060,JAPAN. INT MED CTR JAPAN,RES INST,DEPT CLIN PHARMACOL,TOKYO 162,JAPAN. OTSUKA PHARMACEUT CO LTD,TOKUSHIMA 77101,JAPAN. GUNMA UNIV,SCH MED,DEPT INTERNAL MED,MAEBASHI,GUMMA 371,JAPAN. NCI,MOL CARCINOGENESIS LAB,NIH,BETHESDA,MD 20892. RI yokoi, tsuyoshi/I-7115-2014 NR 29 TC 43 Z9 44 U1 0 U2 0 PU CHAPMAN HALL LTD PI LONDON PA 2-6 BOUNDARY ROW, LONDON, ENGLAND SE1 8HN SN 0960-314X J9 PHARMACOGENETICS JI Pharmacogenetics PD OCT PY 1996 VL 6 IS 5 BP 395 EP 401 DI 10.1097/00008571-199610000-00003 PG 7 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Pharmacology & Pharmacy SC Biotechnology & Applied Microbiology; Genetics & Heredity; Pharmacology & Pharmacy GA VU299 UT WOS:A1996VU29900003 PM 8946471 ER PT J AU Aulakh, CS MazzolaPomietto, P Murphy, DL AF Aulakh, CS MazzolaPomietto, P Murphy, DL TI Long-Term antidepressant treatment restores clonidine's effect on growth hormone secretion in a genetic animal model of depression SO PHARMACOLOGY BIOCHEMISTRY AND BEHAVIOR LA English DT Article DE imipramine; desipramine; fluoxetine; clorgyline; fawn-hooded rat strain; yohimbine ID FAWN-HOODED RATS; NEURO-ENDOCRINE RESPONSES; SPRAGUE-DAWLEY RATS; M-CHLOROPHENYLPIPERAZINE; LITHIUM TREATMENT; RELEASE; WISTAR; HYPOPHAGIA; PREFERENCE; RECEPTORS AB We have recently demonstrated that various doses of clonidine failed to increase growth hormone (GH) in Fawn-hooded (FH) rats (a rat strain suggested to represent a genetic model of depression). In the present study, we investigated whether long-term antidepressant treatment might normalize clonidine's effect on GH secretion in male FH rats. Long-term (16 days) treatment with the tricyclic antidepressant, imipramine (5 mg/kg/day), the 5-HT uptake inhibiting antidepressant, fluoxetine (2.5 mg/kg/day), and the noradrenergic uptake inhibiting antidepressant, desipramine (5 mg/kg/day), accentuated clonidine's effect on GH levels. On the other hand, long-term treatment with the monoamine oxidase type-A inhibiting antidepressant, clorgyline (1 mg/kg/day) and the alpha(2)-noradrenergic antagonists, yohimbine and 1-phenylpiperazine (1 mg/kg/day, each) did not modify clonidine's effect. These findings suggest enhancement of 5-HT2C receptor-mediated function following long-term treatment with uptake inhibiting antidepressants in a genetic animal model of depression. Copyright (C) 1996 Elsevier Science Inc. RP Aulakh, CS (reprint author), NIMH,CLIN SCI LAB,10 CTR DR,MSC 1264,BETHESDA,MD 20892, USA. NR 24 TC 9 Z9 9 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, ENGLAND OX5 1GB SN 0091-3057 J9 PHARMACOL BIOCHEM BE JI Pharmacol. Biochem. Behav. PD OCT PY 1996 VL 55 IS 2 BP 265 EP 268 DI 10.1016/S0091-3057(96)00080-9 PG 4 WC Behavioral Sciences; Neurosciences; Pharmacology & Pharmacy SC Behavioral Sciences; Neurosciences & Neurology; Pharmacology & Pharmacy GA VV761 UT WOS:A1996VV76100013 PM 8951963 ER PT J AU Crouch, RK Chader, GJ Wiggert, B Pepperberg, DR AF Crouch, RK Chader, GJ Wiggert, B Pepperberg, DR TI Retinoids and the visual process SO PHOTOCHEMISTRY AND PHOTOBIOLOGY LA English DT Review ID RETINAL-PIGMENT EPITHELIUM; BINDING PROTEIN IRBP; ALL-TRANS-RETINOIDS; ROD OUTER SEGMENTS; VITAMIN-A; BOVINE RHODOPSIN; CONSTITUTIVE ACTIVATION; BLEACHING ADAPTATION; MICROSOMAL PROTEIN; VERTEBRATE RETINA C1 MED UNIV S CAROLINA,DEPT BIOCHEM,CHARLESTON,SC 29425. NEI,RETINAL CELL & MOL BIOL LAB,BETHESDA,MD 20892. UNIV ILLINOIS,COLL MED,DEPT OPHTHALMOL & VISUAL SCI,CHICAGO,IL. RP Crouch, RK (reprint author), MED UNIV S CAROLINA,DEPT OPHTHALMOL,171 ASHLEY AVE,CHARLESTON,SC 29425, USA. FU NEI NIH HHS [EY04939, EY05494] NR 114 TC 81 Z9 81 U1 0 U2 4 PU AMER SOC PHOTOBIOLOGY PI AUGUSTA PA BIOTECH PARK, 1021 15TH ST, SUITE 9, AUGUSTA, GA 30901-3158 SN 0031-8655 J9 PHOTOCHEM PHOTOBIOL JI Photochem. Photobiol. PD OCT PY 1996 VL 64 IS 4 BP 613 EP 621 DI 10.1111/j.1751-1097.1996.tb03114.x PG 9 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA VM136 UT WOS:A1996VM13600001 PM 8863467 ER PT J AU Bruggemann, E Handwerger, K Essex, C Storz, G AF Bruggemann, E Handwerger, K Essex, C Storz, G TI Analysis of fast neutron-generated mutants at the Arabidopsis thaliana HY4 locus SO PLANT JOURNAL LA English DT Article ID REPRESENTATIONAL DIFFERENCE ANALYSIS; BLUE-LIGHT PHOTORECEPTOR; HYPOCOTYL ELONGATION; POLYMORPHIC MARKERS; GENOMIC SUBTRACTION; GENE; DNA; MUTATIONS; DELETION; CLONING AB Ionizing radiation is expected to produce mutants with deletions or other chromosomal rearrangements. These mutants are useful for a variety of purposes, such as creating null alleles and cloning genes whose existence is known only from their mutant phenotype; however, only a few mutations generated by ionizing radiation have been characterized at the molecular level in Arabidopsis thaliana. Twenty fast neutron-generated alleles of the Arabidopsis HY4 locus, which encodes a blue light receptor, CRY1, were isolated and characterized. Nine of the mutant alleles displayed normal genetic behavior. The other 11 mutant alleles were poorly transmitted through the male gametophyte and were lethal in homozygous plants. Southern blot analysis demonstrated that alleles of the first group generally contain small or moderate-sized deletions at HY4, while alleles of the second group contain large deletions at this locus. These results demonstrate that fast neutrons can produce a range of deletions at a single locus in Arabidopsis. Many of these deletions would be suitable for cloning by genomic subtraction or representational difference analysis. The results also suggest the presence of an essential locus adjacent to HY4. C1 NICHHD,CELL BIOL & METAB BRANCH,NIH,BETHESDA,MD 20892. OI Storz, Gisela/0000-0001-6698-1241 NR 23 TC 68 Z9 73 U1 1 U2 4 PU BLACKWELL SCIENCE LTD PI OXFORD PA OSNEY MEAD, OXFORD, OXON, ENGLAND OX2 0EL SN 0960-7412 J9 PLANT J JI Plant J. PD OCT PY 1996 VL 10 IS 4 BP 755 EP 760 DI 10.1046/j.1365-313X.1996.10040755.x PG 6 WC Plant Sciences SC Plant Sciences GA VN087 UT WOS:A1996VN08700018 PM 8893551 ER PT J AU Gnarra, JR Zhou, SB Merrill, MJ Wagner, JR Krumm, A Papavassiliou, E Oldfield, EH Klausner, RD Linehan, WM AF Gnarra, JR Zhou, SB Merrill, MJ Wagner, JR Krumm, A Papavassiliou, E Oldfield, EH Klausner, RD Linehan, WM TI Post-transcriptional regulation of vascular endothelial growth factor mRNA by the product of the VHL tumor suppressor gene SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID VONHIPPEL-LINDAU DISEASE; MESSENGER-RNA STABILITY; RENAL-CARCINOMA; SOMATIC MUTATIONS; EXPRESSION; ANGIOGENESIS; ELONGATION; CELLS; PROTEIN; HYPOXIA AB The VHL tumor suppressor gene is inactivated in patients with von Hippel-Lindau disease and in most sporadic clear cell renal carcinomas. Although VHL protein function remains unclear, VHL does interact with the elongin BC subunits in vivo and regulates RNA polymerase II elongation activity in vitro by inhibiting formation of the elongin ABC complex, Expression of wild-type VHL in renal carcinoma cells with inactivated endogenous VHL resulted in unaltered in vitro cell growth and decreased vascular endothelial growth factor (VEGF) mRNA expression and responsiveness to serum deprivation. VEGF is highly expressed in many tumors, including VHL-associated and sporadic renal carcinomas, and it stimulates neoangiogenesis in growing solid tumors. Despite 5-fold differences in VEGF mRNA levels, VHL overexpression did not affect VEGF transcription initiation or elongation as would have been suggested by VHL-elongin association. These results suggest that VHL regulates VEGF expression at a post-transcriptional level and that VHL inactivation in target cells causes a loss of VEGF suppression, leading to formation of a vascular stroma. C1 NCI,SURG BRANCH,UROL ONCOL SECT,BETHESDA,MD 20892. NCI,OFF DIRECTOR,BETHESDA,MD 20892. NINCDS,SURG NEUROL BRANCH,NIH,BETHESDA,MD 20892. FRED HUTCHINSON CANC RES CTR,SEATTLE,WA 98104. NR 43 TC 404 Z9 420 U1 1 U2 1 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD OCT 1 PY 1996 VL 93 IS 20 BP 10589 EP 10594 DI 10.1073/pnas.93.20.10589 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VL333 UT WOS:A1996VL33300012 PM 8855222 ER PT J AU Rovescalli, AC Asoh, S Nirenberg, M AF Rovescalli, AC Asoh, S Nirenberg, M TI Cloning and characterization of four murine homeobox genes SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE mouse embryo; transcription factors; Uncx-4.1; OG-2; OG-9; OG-12 ID HOMEODOMAIN PROTEIN; SUBSTRATE RECOGNITION; SYNAPTIC INPUT; MOTOR NEURONS; DROSOPHILA; SPECIFICITY; BINDING; KINASE; EXPRESSION; ENCODES AB Four novel murine homeobox genes, Uncx-4.1, OG-2, OG-9, and OG-12, were cloned and partially sequenced, The amino acid sequence of the mouse Uncx-4.1 homeodomain is closely related to the sequence of the unc-4 homeodomain of Caenorhabditis elegans, However, the OG-2, OG-9, and OG-12 homeodomains are relatively diverged and are not closely related to any previously described homeodomain, Northern blot analyses revealed multiple bands of Uncx-4.1, OG-2, OG-9, and OG-12 poly(A)(+) RNA in RNA from mouse embryos and adults that change during development and showed that each gene is expressed in a tissue-specific manner, OG-12 cDNAs were cloned that correspond to two alternatively spliced species of OG-12 mRNA, Three major bands of Uncx-4.1 poly(A)(+) RNA were found only in RNA from adult mouse brain, but an additional band was observed in RNA from all of the other tissues tested, Major bands of OG-9 and OG-2 poly(A)(+) RNA were found only in RNA from striated muscle; however, trace bands were detected in RNA from other tissues. C1 NHLBI, LAB BIOCHEM GENET, NIH, BETHESDA, MD 20892 USA. NR 33 TC 50 Z9 55 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 OCT 1 PY 1996 VL 93 IS 20 BP 10691 EP 10696 DI 10.1073/pnas.93.20.10691 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VL333 UT WOS:A1996VL33300031 PM 8855241 ER PT J AU Hung, CH Srivastava, M Pollard, HB AF Hung, CH Srivastava, M Pollard, HB TI Membrane fusion protein synexin (annexin VII) as a Ca2+/GTP sensor in exocytotic secretion SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID ADRENAL CHROMAFFIN CELLS; CALCIUM-CHANNEL ACTIVITY; GTP-BINDING PROTEIN; MAST-CELLS; NEUROTRANSMITTER RELEASE; CATECHOLAMINE RELEASE; REGULATED EXOCYTOSIS; MOLECULAR MECHANISM; ADP-RIBOSYLATION; GRANULE GHOSTS AB Exocytotic membrane fusion and secretion are promoted by the concerted action of GTP and Ca2+, although the precise site(s) of action in the process are not presently known, However, the calcium-dependent membrane fusion reaction driven by synexin (annexin VII) is an in vitro model for this process, which we have now found to be further activated by GTP, The mechanism of fusion activation depends on the unique ability of synexin to bind and hydrolyze GTP in a calcium-dependent manner, both in vitro and in vivo in streptolysin O-permeabilized chromaffin cells, The required [Ca2+] for GTP binding by synexin is in the range of 50-200 mu M, which is known to occur at exocytotic sites in chromaffin cells, neurons, and other cell types, Previous immunolocalization studies place synexin at exocytotic sites in chromaffin cells, and we conclude that synexin is an atypical G protein that may be responsible for both detecting and mediating the Ca2+/GTP signal for exocytotic membrane fusion. C1 NIDDKD, CELL BIOL & GENET LAB, NIH, BETHESDA, MD 20892 USA. NR 75 TC 2 Z9 2 U1 1 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 OCT 1 PY 1996 VL 93 IS 20 BP 10797 EP 10802 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VL333 UT WOS:A1996VL33300050 ER PT J AU Kwak, LW Young, HA Pennington, RW Weeks, SD AF Kwak, LW Young, HA Pennington, RW Weeks, SD TI Vaccination with syngeneic, lymphoma-derived immunoglobulin idiotype combined with granulocyte/macrophage colony-stimulating factor primes mice for a protective T-cell response SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE cancer vaccines; cytokines; myeloma ID CLASS-II MOLECULES; BONE-MARROW; ANTITUMOR IMMUNITY; MYELOMA PROTEINS; TUMOR-CELLS; IMMUNIZATION; INDUCTION; ANTIGENS; MACROPHAGES; ANTIBODIES AB The idiotype of the Ig expressed by a B-cell malignancy (Id) can serve as a unique tumor-specific antigen and as a model for cancer vaccine development, In murine models of Id vaccination, formulation of syngeneic Id with carrier proteins or adjuvants induces an anti-idiotypic antibody response, However, inducing a potent cell-mediated response to this weak antigen instead would be highly desirable, In the 38C13 lymphoma model, we observed that low doses of free granulocyte/macrophage colony-stimulating factor (GM-CSF) 10,000 units i.p. or locally s.c. daily for 4 days significantly enhanced protective antitumor immunity induced by s.c. Id-keyhole limpet hemocyanin (KLH) immunization, This effect was critically dependent upon effector CD4(+) and CD8(+) T cells and was not associated with any increased anti-idiotypic antibody production, Lymphocytes from spleens and draining lymph nodes of mice primed with Id-KLH plus GM-CSF, but not with Id-KLH alone, demonstrated significant proliferation to Id in vitro without any biased production of interferon gamma or interleukin 4 protein or mRNA, As a further demonstration of potency, 50% of mice immunized with Id-KLH plus GM-CSF on the same day as challenge with a large s.c. tumor inoculum remained tumor-free at day 80, compared with 17% for Id-KLH alone, when immunization was combined with cyclophosphamide. Taken together, these results demonstrate that GM-CSF can significantly enhance the immunogenicity of a defined self-antigen and that this effect is mediated exclusively by activating the T cell arm of the immune response. C1 NCI,EXPT IMMUNOL LAB,FREDERICK,MD 21702. SAIC,INTRAMURAL RES SUPPORT PROGRAM,FREDERICK,MD 21702. RP Kwak, LW (reprint author), NCI,DIV CLIN SCI,FREDERICK,MD 21702, USA. NR 36 TC 184 Z9 191 U1 0 U2 3 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD OCT 1 PY 1996 VL 93 IS 20 BP 10972 EP 10977 DI 10.1073/pnas.93.20.10972 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VL333 UT WOS:A1996VL33300083 PM 8855293 ER PT J AU Shimojo, N Kohno, Y Yamaguchi, K Kikuoka, S Hoshioka, A Niimi, H Hirai, A Tamura, Y Saito, Y Kohn, LD Tahara, K AF Shimojo, N Kohno, Y Yamaguchi, K Kikuoka, S Hoshioka, A Niimi, H Hirai, A Tamura, Y Saito, Y Kohn, LD Tahara, K TI Induction of Graves-like disease in mice by immunization with fibroblasts transfected with the thyrotropin receptor and a class II molecule SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID SYSTEMIC LUPUS-ERYTHEMATOSUS; AUTOIMMUNE THYROID-DISEASE; MONOCLONAL-ANTIBODIES; EXTRACELLULAR DOMAIN; TSH BINDING; EXPRESSION; HLA; CELLS; HYPERTHYROXINEMIA; IMMUNOGLOBULINS AB Graves disease is an autoimmune thyroid disease characterized by the presence of antibodies against the thyrotropin receptor (TSHR), which stimulate the thyroid to cause hyperthyroidism and/or goiter. By immunizing mice with fibroblasts transfected with both the human TSHR and a major histocompatibility complex class II molecule, but not by either alone, we have induced immune hyperthyroidism that has the major humoral and histological features of Graves disease: stimulating TSHR antibodies, thyrotropin binding inhibiting immunoglobulins, which are different from the stimulating TSHR antibodies, increased thyroid hormone levels, thyroid enlargement, thyrocyte hypercellularity, and thyrocyte intrusion into the follicular lumen. The results suggest that the aberrant expression of major histocompatibility complex class II molecules on cells that express a native form of the TSHR can result in the induction of functional anti-TSHR antibodies that stimulate the thyroid. They additionally suggest that the acquisition of antigen-presenting ability on a target cell containing the TSHR can activate T and B cells normally present in an animal and induce a disease with the major features of autoimmune Graves. C1 CHIBA UNIV,SCH MED,DEPT PEDIAT,CHIBA 260,JAPAN. CHIBA UNIV,SCH MED,DEPT INTERNAL MED 2,CHIBA 260,JAPAN. NIH,NIDDK,BETHESDA,MD 20892. NR 33 TC 163 Z9 179 U1 0 U2 1 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD OCT 1 PY 1996 VL 93 IS 20 BP 11074 EP 11079 DI 10.1073/pnas.93.20.11074 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VL333 UT WOS:A1996VL33300101 PM 8855311 ER PT J AU Bozza, PT Payne, JL Morham, SG Langenbach, R Smithies, O Weller, PF AF Bozza, PT Payne, JL Morham, SG Langenbach, R Smithies, O Weller, PF TI Leukocyte lipid body formation and eicosanoid generation: Cyclooxygenase-independent inhibition by aspirin SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE nonsteroidal antiinflammatory agents; cyclooxygenase knockout; leukotrienes; prostaglandin endoperoxide H synthase; lipoxygenase ID ENDOPEROXIDE SYNTHASE CYCLOOXYGENASE; PROSTAGLANDIN-G/H SYNTHASE; PROTEIN-KINASE-C; ARACHIDONIC-ACID; POLYMORPHONUCLEAR LEUKOCYTES; ALVEOLAR MACROPHAGES; HUMAN EOSINOPHILS; HUMAN-NEUTROPHILS; GENE DISRUPTION; BODIES AB Lipid bodies, cytoplasmic inclusions that develop in cells associated with inflammation, are inducible structures that might participate in generating inflammatory eicosanoids, Cis-unsaturated fatty acids (arachidonic and oleic acids) rapidly induced lipid body formation in leukocytes, and this lipid body induction was inhibited by aspirin and nonsteroidal antiinflammatory drugs (NSAIDs), Several findings indicated that the inhibitory effect of aspirin and NSAIDs on lipid body formation was independent of cyclooxygenase (COX) inhibition, First, the non-COX inhibitor, sodium salicylate, was as potent as aspirin in inhibiting lipid body formation elicited by cis-fatty acids, Second, cis-fatty acid-induced lipid body formation was not impaired in macrophages from COX-1 or COX-2 genetically deficient mice, Finally, NSAIDs inhibited arachidonic acid-induced lipid body formation likewise in macrophages from wild-type and COX-1- and COX-2-deficient mice, An enhanced capacity to generate eicosanoids developed after 1 hr concordantly with cis-fatty acid-induced lipid body formation, Arachidonic and oleic acid-induced lipid body numbers correlated with the enhanced levels of leukotrienes B-4 and C-4 and prostaglandin E(2) produced after submaximal calcium ionophore stimulation, Aspirin and NSAIDs inhibited both induced lipid body formation and the enhanced capacity for forming leukotrienes as well as prostaglandins. Our studies indicate that lipid body formation is an inducible early response in leukocytes that correlates with enhanced eicosanoid synthesis, Aspirin and NSAIDs, independent of COX inhibition, inhibit cis-fatty acid-induced lipid body formation in leukocytes and in concert inhibit the enhanced synthesis of leukotrienes and prostaglandins. C1 HARVARD UNIV,BETH ISRAEL HOSP,DEPT MED,SCH MED,CHARLES A DANA RES INST,BOSTON,MA 02215. BETH ISRAEL HOSP,HARVARD THORNDIKE LAB,BOSTON,MA 02215. UNIV N CAROLINA,DEPT PATHOL,CHAPEL HILL,NC 27599. NIEHS,LAB EXPT CARCINOGENESIS & MUTAGENESIS,RES TRIANGLE PK,NC 27709. OI Bozza, Patricia/0000-0001-8349-9529 FU NIAID NIH HHS [AI 22571] NR 40 TC 85 Z9 85 U1 0 U2 2 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD OCT 1 PY 1996 VL 93 IS 20 BP 11091 EP 11096 DI 10.1073/pnas.93.20.11091 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VL333 UT WOS:A1996VL33300104 PM 8855314 ER PT J AU Orlic, D Girard, LJ Jordan, CT Anderson, SM Cline, AP Bodine, DM AF Orlic, D Girard, LJ Jordan, CT Anderson, SM Cline, AP Bodine, DM TI The level of mRNA encoding the amphotropic retrovirus receptor in mouse and human hematopoietic stem cells is low and correlates with the efficiency of retrovirus transduction SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID HUMAN ADENOSINE-DEAMINASE; APE LEUKEMIA-VIRUS; MEDIATED GENE-TRANSFER; BONE-MARROW CELLS; LONG-TERM EXPRESSION; UMBILICAL-CORD BLOOD; COLONY-STIMULATING FACTOR; PERIPHERAL-BLOOD; PROGENITOR CELLS; MURINE RETROVIRUSES AB The low level of amphotropic retrovirus-mediated gene transfer into human hematopoietic stem cells (HSC) has been a major impediment to gene therapy for hematopoietic diseases, In the present study, we have examined amphotropic retrovirus receptor (amphoR) and ecotropic retrovirus receptor mRNA expression in highly purified populations of mouse and human HSC. Murine HSC with low to undetectable levels of amphoR mRNA and relatively high levels of ecotropic retrovirus receptor mRNA were studied, When these HSC were analyzed simultaneously for ecotropic and amphotropic retrovirus transduction, ecotropic provirus sequences were detected in 10 of 13 long-term repopulated animals, while amphotropic proviral sequences were detected in only one recipient, A second distinct population of murine HSC were isolated that express 3-fold higher levels of amphoR mRNA, When these HSC were analyzed simultaneously for ecotropic and amphotropic retrovirus transduction, 11 of 11 repopulated mice contained ecotropic provirus and 6 of 11 contained amphotropic provirus sequences, a significant increase in the amphotropic retrovirus transduction (P = 0.018), These results indicate that, among the heterogeneous populations of HSC present in adult mouse bone marrow, the subpopulation with the highest level of amphoR mRNA is more efficiently transduced by amphotropic retrovirus, In a related study, we found low levels of human amphoR mRNA in purified populations of human HSC (CD34(+) CD38(-)) and higher levels in committed progenitor cells (CD34(+) CD38(+)). We conclude that the amphoR mRNA level in HSC correlates with amphotropic retrovirus transduction efficiency. C1 SOMATIX THERAPY CORP,ALAMEDA,CA 94501. RP Orlic, D (reprint author), NIH,HEMATOPOIESIS SECT,LAB GENE TRANSFER,NATL CTR HUMAN GENOME RES,49 CONVENT DR,ROOM 3A14,BETHESDA,MD 20892, USA. NR 47 TC 192 Z9 193 U1 0 U2 1 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD OCT 1 PY 1996 VL 93 IS 20 BP 11097 EP 11102 DI 10.1073/pnas.93.20.11097 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VL333 UT WOS:A1996VL33300105 PM 8855315 ER PT J AU Su, H Raymond, L Rockey, DD Fischer, E Hackstadt, T Caldwell, HD AF Su, H Raymond, L Rockey, DD Fischer, E Hackstadt, T Caldwell, HD TI A recombinant Chlamydia trachomatis major outer membrane protein binds to heparan sulfate receptors on epithelial cells SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE chlamydiae; pathogenesis; cytoadhesin; glycosaminoglycan receptor ID HERPES-SIMPLEX VIRUS; FIBROBLASTS L-CELLS; HELA-CELLS; SURFACE; PSITTACI; PROTEOGLYCANS; FAMILY; BIOSYNTHESIS; ATTACHMENT; INHIBITION AB Chlamydial attachment to columnar conjunctival or urogenital epithelial cells is an initial and critical step in the pathogenesis of chlamydial mucosal infections, The chlamydial major outer membrane protein (MOMP) has been implicated as a putative chlamydial cytoadhesin; however, direct evidence supporting this hypothesis has not been reported, The function of MOMP as a cytoadhesin was directly investigated by expressing the protein as a fusion with the Escherichia coli maltose binding protein (MBP-MOMP) and studying its interaction with human epithelial cells, The recombinant MBP-MOMP bound specifically to HeLa cells at 4 degrees C but was not internalized after shifting the temperature to 37 degrees C. The MBP-MOMP competitively inhibited the infectivity of viable chlamydiae for epithelial cells, indicating that the MOMP and intact chlamydiae bind the same host receptor, Heparan sulfate markedly reduced binding of the MBP-MOMP to cells, whereas chondroitin sulfate had no effect on binding, Enzymatic treatment of cells with heparitinase but not chondroitinase inhibited the binding of MBP-MOMP. These same treatments were also shown to reduce the infectivity of chlamydiae for epithelial cells, Mutant cell lines defective in heparan sulfate synthesis but not chondroitin sulfate synthesis showed a marked reduction in the binding of MBP-MOMP and were also less susceptible to infection by chlamydiae. Collectively, these findings provide strong evidence that the MOMP functions as a chlamydial cytoadhesin and that heparan sulfate proteoglycans are the host-cell receptors to which the MOMP binds. C1 NIAID,ROCKY MT LAB,INTRACELLULAR PARASITES LAB,HAMILTON,MT 59840. NIAID,ROCKY MT LAB,MICROSCOPY BRANCH,HAMILTON,MT 59840. NR 34 TC 114 Z9 115 U1 1 U2 11 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD OCT 1 PY 1996 VL 93 IS 20 BP 11143 EP 11148 DI 10.1073/pnas.93.20.11143 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VL333 UT WOS:A1996VL33300113 PM 8855323 ER PT J AU Ohshima, T Ward, JM Huh, CG Longenecker, G Veeranna Pant, HC Brady, RO Martin, LJ Kulkarni, AB AF Ohshima, T Ward, JM Huh, CG Longenecker, G Veeranna Pant, HC Brady, RO Martin, LJ Kulkarni, AB TI Targeted disruption of the cyclin-dependent kinase 5 gene results in abnormal corticogenesis, neuronal pathology and perinatal death SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID DIRECTED PROTEIN-KINASE; GANGLION-CELL NEURONS; NEUROFILAMENT PROTEINS; POSTTRANSLATIONAL MODIFICATION; MAMMALIAN NEUROFILAMENTS; PHOSPHORYLATION SITES; CDC2-LIKE KINASE; AXONAL-TRANSPORT; BRAIN; TAU AB Although cyclin-dependent kinase 5 (Cdk5) is closely related to other cyclin-dependent kinases, its kinase activity is detected only in the postmitotic neurons. Cdk5 expression and kinase activity are correlated with the extent of differentiation of neuronal cells in developing brain, Cdk5 purified from nervous tissue phosphorylates neuronal cytoskeletal proteins including neurofilament proteins and microtubule-associated protein tau in vitro. These findings indicate that Cdk5 may have unique functions in neuronal cells, especially in the regulation of phosphorylation of cytoskeletal molecules, We report here generation of Cdk5(-/-) mice through gene targeting and their phenotypic analysis. Cdk5(-/-) mice exhibit unique lesions in the central nervous system associated with perinatal mortality. The brains of Cdk5(-/-) mice lack cortical laminar structure and cerebellar foliation, In addition, the large neurons in the brain stem and in the spinal cord show chromatolytic changes with accumulation of neurofilament immunoreactivity. These findings indicate that Cdk5 is an important molecule for brain development and neuronal differentiation and also suggest that Cdk5 may play critical roles in neuronal cytoskeleton structure and organization. C1 NIDA,GENE TARGETING RES & CORE FACIL,NIH,BETHESDA,MD 20892. NINCDS,DEV & METAB NEUROL BRANCH,NIH,BETHESDA,MD 20892. NINCDS,NEUROCHEM LAB,NIH,BETHESDA,MD 20892. NCI,VET & TUMOR PATHOL SECT,OFF LAB ANIM SCI,NIH,FREDERICK,MD 21702. JOHNS HOPKINS UNIV,SCH MED,DEPT PATHOL,NEUROPATHOL LAB,BALTIMORE,MD 21205. NR 45 TC 665 Z9 686 U1 1 U2 3 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD OCT 1 PY 1996 VL 93 IS 20 BP 11173 EP 11178 DI 10.1073/pnas.93.20.11173 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VL333 UT WOS:A1996VL33300118 PM 8855328 ER PT J AU Kitzler, JW Eling, TE AF Kitzler, JW Eling, TE TI Cloning, sequencing and expression of a 5-lipoxygenase from Syrian hamster embryo fibroblasts SO PROSTAGLANDINS LEUKOTRIENES AND ESSENTIAL FATTY ACIDS LA English DT Article ID AMINO-ACID SEQUENCE; MOLECULAR-CLONING; ARACHIDONIC-ACID; DIETARY-CHOLESTEROL; CDNA CLONING; LIPOXYGENASES; 15-LIPOXYGENASE; METABOLITES; MUTAGENESIS; 12-LIPOXYGENASE AB The hamster ortholog of human and rat 5-lipoxygenase (5-LO) was cloned from a Syrian hamster embryo (SHE) cell line. A combination of polymerase chain reaction (PCR) and 5' and 3' RACE (rapid amplification of cDNA ends) was used to isolate the complete cDNA for this gene. The cDNA sequence demonstrates the extreme sequence conservation found in this gene family, with a deduced amino acid sequence 95% identical to the rat 5-LO, and 90% identical to the human enzyme. The hamster 5-LO was expressed in E. coli The expressed protein was detected by an antibody to human 5-LO, and had an apparent molecular weight of 75-80 kD. The products of the action of this enzyme on arachidonic acid are 5-HETE and the diHETEs resulting from the breakdown of LTA(4), in a pattern similar to that produced by the recombinant human 5-LO. No oxidation of linoleic acid by this enzyme was detected. C1 NIEHS,NIH,MOL BIOPHYS LAB,RES TRIANGLE PK,NC 27709. NR 29 TC 9 Z9 10 U1 0 U2 0 PU CHURCHILL LIVINGSTONE PI EDINBURGH PA JOURNAL PRODUCTION DEPT, ROBERT STEVENSON HOUSE, 1-3 BAXTERS PLACE, LEITH WALK, EDINBURGH, MIDLOTHIAN, SCOTLAND EH1 3AF SN 0952-3278 J9 PROSTAG LEUKOTR ESS JI Prostaglandins Leukot. Essent. Fatty Acids PD OCT PY 1996 VL 55 IS 4 BP 269 EP 277 DI 10.1016/S0952-3278(96)90008-3 PG 9 WC Biochemistry & Molecular Biology; Cell Biology; Endocrinology & Metabolism SC Biochemistry & Molecular Biology; Cell Biology; Endocrinology & Metabolism GA VU431 UT WOS:A1996VU43100009 PM 8951996 ER PT J AU Zdanov, A SchalkHihi, C Wlodawer, A AF Zdanov, A SchalkHihi, C Wlodawer, A TI Crystal structure of human interleukin-10 at 1.6 angstrom resolution and a model of a complex with its soluble receptor SO PROTEIN SCIENCE LA English DT Article DE cytokines; four-helix-bundle; interferon gamma; interleukin-10; receptor binding ID EPSTEIN-BARR-VIRUS; INTERFERON-GAMMA; EXTRACELLULAR DOMAIN; CYTOKINE SYNTHESIS; HEXAMERIC COMPLEX; GROWTH-HORMONE; CELLS; IL-10; EXPRESSION; BINDING AB The crystal structure of human interleukin-10 (IL-10) was refined at 1.6 Angstrom resolution against X-ray diffraction data collected at 100 K with the use of synchrotron radiation. Although similar to the IL-10 structure determined previously at room temperature, this low-temperature IL-10 structure contains, in addition, four N-terninal residues, three sulfate anions, and 175 extra water molecules. Whereas the main-chain conformation is preserved, about 30% of the side chains, most of them on the protein surface, assume different conformations. A computer model of a complex of IL-10 with its two soluble receptors was generated based on the topological similarity of IL-10 to interferon-gamma. The contact region between the cytokine and each receptor shows excellent complementarity of polar and hydrophobic interactions, suggesting that the model is generally correct and should be useful in guiding mutagenesis experiments. C1 NCI,MACROMOL STRUCT LAB,FREDERICK CANC RES & DEV CTR,ABL BASIC RES PROGRAM,FREDERICK,MD 21702. NR 39 TC 76 Z9 79 U1 0 U2 3 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 SN 0961-8368 J9 PROTEIN SCI JI Protein Sci. PD OCT PY 1996 VL 5 IS 10 BP 1955 EP 1962 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VL922 UT WOS:A1996VL92200001 PM 8897595 ER PT J AU Wolff, J Sackett, DL Knipling, L AF Wolff, J Sackett, DL Knipling, L TI Cation selective promotion of tubulin polymerization by alkali metal chlorides SO PROTEIN SCIENCE LA English DT Article DE cation selective charge shielding; microtubule-associated proteins; tubulin polymerization rate; tubulin S; urea-induced depolymerization ID MICROTUBULE-ASSOCIATED PROTEINS; HELICAL COILED-COILS; ELECTROSTATIC INTERACTIONS; INVITRO; BINDING; BRAIN; REPULSIONS; STABILITY; DIMER; MAP2 AB A role for charge-based interactions in protein stability at the monomer or dimer level is well known. We show here that such interactions can also be important for the higher-order structures of microtubule assembly. Alkali metal chlorides increase the rate of polymerization of pure tubulin driven by either taxol or dimethyl sulfoxide. The effect is cation selective, exhibiting a sequence Na+ > K+ > Li+ > Cs+, with optimal concentrations for Na+ at similar to 160 mM. Hofmeister anion effects are additive with these rate stimulations. Sodium is less potent than guanidinium ion stimulation reported previously, but produces a larger fraction of normal microtubules. Alkali metal cations lower the critical concentration by a factor of similar to 2, produce cold reversible polymers whose formation is sensitive to podophyllotoxin inhibition, increase the fraction of polymers present as microtubules from similar to 0.9 to 0.99, and reverse or prevent urea-induced depolymerization of microtubules. In the presence of microtubule-associated proteins, the promotion of polymerization is no longer cation selective. In the polymerization of tubulin S, in which the acidic C termini of both monomers have been cleaved, the cation enhancement is markedly decreased, although selectivity persists. Because the selectivity sequence is similar to that of the coil/helix transition of polyglutamic acid, we suggest that a major part, although not all, of the cation selective enhancement of polymerization results from shielding of the glutamate-rich C termini of the tubulin monomers. RP Wolff, J (reprint author), NIDDKD,BIOCHEM PHARMACOL LAB,NIH,BLDG 8,BETHESDA,MD 20892, USA. NR 58 TC 50 Z9 51 U1 0 U2 4 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 40 WEST 20TH STREET, NEW YORK, NY 10011-4211 SN 0961-8368 J9 PROTEIN SCI JI Protein Sci. PD OCT PY 1996 VL 5 IS 10 BP 2020 EP 2028 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VL922 UT WOS:A1996VL92200008 PM 8897602 ER PT J AU Bryant, SH AF Bryant, SH TI Evaluation of threading specificity and accuracy SO PROTEINS-STRUCTURE FUNCTION AND GENETICS LA English DT Article DE contact potentials; fold recognition; protein threading; Gibbs sampling ID PROTEIN-STRUCTURE PREDICTION; SIDE-CHAIN PACKING; CRYSTAL-STRUCTURE; FOLD RECOGNITION; SEQUENCE; PHYCOCYANINS; ALIGNMENT; DATABASES; FEATURES; GLOBINS AB Threading experiments with proteins from the globin family provide an indication of the nature of the structural similarity required for successful fold recognition and accurate sequence-structure alignment. Threading scores are fouled to rise above the noise of false positives whenever roughly 60% of residues from a sequence can be aligned with analogous sites in the structure of a remote homolog. Fold recognition specificity thus appears to be limited by the extent of structural similarity, regardless of the degree of sequence similarity. Threading alignment accuracy is found to depend more critically on the degree of structural similarity. Alignments are accurate, placing the majority of residues exactly as in structural alignment, only when superposition residuals are less than 2.5 Angstrom. These criteria for successful recognition and sequence-structure alignment appear to be consistent with the successes and failures of threading methods in blind structure prediction. They also suggest a direct assay for improved threading methods: Potentials and alignment models should be tested for their ability to detect less extensive structural similarities, and to produce accurate alignments when superposition residuals for this conserved ''core'' fall in the range characteristic of remote homologs. (C) 1996 Wiley-Liss, Inc. RP Bryant, SH (reprint author), NIH,COMPUTAT BIOL BRANCH,NATL CTR BIOTECHNOL INFORMAT,8600 ROCKVILLE PIKE,BETHESDA,MD 20894, USA. NR 39 TC 57 Z9 59 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC 605 THIRD AVE, NEW YORK, NY 10158-0012 SN 0887-3585 J9 PROTEINS JI Proteins PD OCT PY 1996 VL 26 IS 2 BP 172 EP 185 PG 14 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA VP791 UT WOS:A1996VP79100007 PM 8916225 ER PT J AU Muntaner, C Stormes, J AF Muntaner, C Stormes, J TI Social class and behavior: Simultaneous class positions yield different amounts of income SO PSYCHOLOGICAL REPORTS LA English DT Article ID SOCIOECONOMIC-STATUS; UNITED-STATES; HEALTH; CHALLENGE AB We used data from the Panel Study of Income Dynamics to present preliminary findings on the social class behavior of the poor and general populations in the USA. Analysis supported the notion that individuals are engaged simultaneously in multiple class positions, e.g., self-employed and welfare recipient, that yield different amounts of income. Moreover, no single label or explanation, e.g., ''underclass'' or ''the poor are marginal to the economy,'' seems appropriate to describe the complexity of the economic behavior of poor individuals. RP Muntaner, C (reprint author), NIMH,LAB SOCIOENVIRONM STUDIES,FED BLDG,ROOM B1A-12,7550 WISCONSIN AVE,BETHESDA,MD 20892, USA. RI Muntaner, C/A-5043-2010 NR 13 TC 5 Z9 5 U1 0 U2 3 PU PSYCHOLOGICAL REPORTS PI MISSOULA PA P O BOX 9229, MISSOULA, MT 59807 SN 0033-2941 J9 PSYCHOL REP JI Psychol. Rep. PD OCT PY 1996 VL 79 IS 2 BP 379 EP 382 PG 4 WC Psychology, Multidisciplinary SC Psychology GA VM337 UT WOS:A1996VM33700006 PM 8909059 ER PT J AU Egan, MF Hurd, Y Ferguson, J Bachus, SE Hamid, EH Hyde, TM AF Egan, MF Hurd, Y Ferguson, J Bachus, SE Hamid, EH Hyde, TM TI Pharmacological and neurochemical differences between acute and tardive vacuous chewing movements induced by haloperidol SO PSYCHOPHARMACOLOGY LA English DT Article DE vacuous chewing movements; haloperidol; tardive dyskinesia; substance P; dynorphin; enkephalin ID PREPROTACHYKININ MESSENGER-RNAS; GLUTAMIC-ACID DECARBOXYLASE; INDUCED ORAL DYSKINESIAS; SUBSTANCE-P; DOPAMINERGIC REGULATION; NEUROLEPTIC TREATMENT; BASAL GANGLIA; STRIATOPALLIDAL NEURONS; STRIATONIGRAL PATHWAY; GENE-EXPRESSION AB Late onset vacuous chewing movements (VCMs) from chronic neuroleptic treatment have been used as a rat model of tardive dyskinesia (TD). Early onset VCMs have also been observed, raising questions about the validity of this model. To assess the relation ship between these two types of VCMs, pharmacological and neurochemical properties of early and late onset VCMs were compared. ''Acute'' VCMs were induced by daily intraperitoneal injections for 1-21 days. ''Tardive'' VCMs were induced by intramuscular injections of haloperidol decanoate every 3 weeks for 30 weeks followed by a 24-week withdrawal period. Suppression was attempted for both types of VCMs using several doses of intraperitoneal haloperidol. Striatonigral activation was assessed by measuring mRNA expression levels of the neuropeptides dynorphin and substance P using in situ hybridization histochemistry. Enkephalin mRNA was also measured as an index of striatopallidal activation. The results indicate that acute VCMs cannot be suppressed with increased doses of haloperidol and are associated with reduced dynorphin and substance P. This profile is similar to that seen with an animal model of parkinsonism. Tardive VCMs, in contrast, were markedly suppressed by haloperidol. They have previously been shown to be associated with increased striatonigral activation as indicated by increased dynorphin mRNA. Enkephalin mRNA was elevated following both short and long term treatment. Although superficially similar, acute and tardive VCMs appear to have different pharmacological and neurochemical profiles, suggesting they are related to acute extrapyramidal side effects and tardive dyskinesia, respectively. C1 ST ELIZABETH HOSP,NIMH,NEUROSCI RES CTR,CLIN BRAIN DISORDERS BRANCH,WASHINGTON,DC 20032. KAROLINSKA HOSP,DEPT PSYCHIAT & PSYCHOL,S-10401 STOCKHOLM,SWEDEN. RP Egan, MF (reprint author), ST ELIZABETH HOSP,NIMH,NEUROSCI RES CTR,CLIN RES SERV,2700 MARTIN LUTHER KING JR AVE SE,WASHINGTON,DC 20032, USA. NR 64 TC 69 Z9 69 U1 1 U2 1 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 SN 0033-3158 J9 PSYCHOPHARMACOLOGY JI Psychopharmacology PD OCT PY 1996 VL 127 IS 4 BP 337 EP 345 DI 10.1007/BF02806012 PG 9 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA VQ024 UT WOS:A1996VQ02400007 PM 8923569 ER PT J AU Avila, NA Mueller, BU Carrasquillo, JA Kontny, HU Jaffe, ES Pizzo, PA AF Avila, NA Mueller, BU Carrasquillo, JA Kontny, HU Jaffe, ES Pizzo, PA TI Multilocular thymic cysts: Imaging features in children with human immunodeficiency virus infection SO RADIOLOGY LA English DT Article DE acquired immunodeficiency syndrome (AIDS); coccidiomycosis; mediastinum, cysts; pneumonitis, lymphocytic interstitial; thymus, cysts ID IMMUNE-DEFICIENCY SYNDROME; GLAND; AIDS AB PURPOSE: To evaluate the radiologic and follow-up features of multilocular thymic cysts in children with human immunodeficiency virus (HIV) infection. MATERIALS AND METHODS: Four HN-infected children with large anterior mediastinal masses depicted at routine chest radiography underwent ultrasonography (US), unenhanced and contrast material-enhanced computed tomography (CT), and unenhanced and gadolinium-enhanced MR imaging of the chest Gallium scanning was also performed in three of the four children. The patients underwent follow-up radiologic examinations for 8-15 months. RESULTS: The multiloculated nature of the masses was depicted at contrast-enhanced but not unenhanced CT. Similarly, the septaions were depicted on T2-weighted, short inversion time inversion-recovery (STIR), and contrast-enhanced T1-weighted MR images but not on the unenhanced T1-weighted images. US scans depicted the septations within each mass, but findings were technically limited because only portions of each mass were depicted. Gallium scans in three masses depicted uptake of radionuclide in two and no uptake in one. Surgical biopsy was performed in each mass: Follicular hyperplasia and diffuse plasmacytosis of the thymus were found but not evidence of neoplastic or infectious origin. At follow-up, the mass decreased in volume in two patients, did not change in one patient, and increased in volume in one patient CONCLUSION: HIV-infected patients with asymptomatic mediastinal masses depicted at routine chest radiography should undergo contrast-enhanced CT. If a solid mass is depicted, biopsy should be performed to exclude neoplastic or infectious origins. If a multiloculated anterior mediastinal mass is depicted, symptomatic follow-up is adequate since the finding represents a rare multilocular thymic cyst that does not have negative clinical implications. C1 NCI,DEPT NUCL MED,NIH,BETHESDA,MD 20892. NCI,WARREN G MAGNUSON CLIN CTR,NIH,BETHESDA,MD 20892. NCI,DIV CLIN SCI,PEDIAT BRANCH,NIH,BETHESDA,MD 20892. NCI,DIV CANC BIOL DIAG & CTR,NIH,BETHESDA,MD 20892. RP Avila, NA (reprint author), NCI,DEPT DIAGNOST RADIOL,NIH,BLDG 10,RM 1C-660,10 CTR DR,BETHESDA,MD 20892, USA. RI Carrasquillo, Jorge/E-7120-2010; OI Carrasquillo, Jorge/0000-0002-8513-5734 NR 16 TC 19 Z9 20 U1 0 U2 0 PU RADIOLOGICAL SOC NORTH AMER PI EASTON PA 20TH AND NORTHAMPTON STS, EASTON, PA 18042 SN 0033-8419 J9 RADIOLOGY JI Radiology PD OCT PY 1996 VL 201 IS 1 BP 130 EP 134 PG 5 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA VJ114 UT WOS:A1996VJ11400029 PM 8816533 ER PT J AU Holland, SM AF Holland, SM TI Therapy of mycobacterial infections SO RESEARCH IN IMMUNOLOGY LA English DT Article ID TUMOR-NECROSIS-FACTOR; AVIUM COMPLEX INFECTION; RECOMBINANT INTERFERON-GAMMA; ACTIVATES HUMAN MACROPHAGES; CELL-MEDIATED-IMMUNITY; LEPROMATOUS LEPROSY; HUMAN-MONOCYTES; FACTOR-ALPHA; INTRACELLULARE COMPLEX; PULMONARY TUBERCULOSIS RP Holland, SM (reprint author), NIAID,HOST DEF LAB,NIH,BLDG 10,11N103,10 CTR DR,MSC 1886,BETHESDA,MD 20892, USA. NR 79 TC 6 Z9 7 U1 0 U2 1 PU EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER PI PARIS PA 141 RUE JAVEL, 75747 PARIS, FRANCE SN 0923-2494 J9 RES IMMUNOL JI Res. Immunol. PD OCT-DEC PY 1996 VL 147 IS 8-9 BP 572 EP 581 DI 10.1016/S0923-2494(97)85224-8 PG 10 WC Immunology SC Immunology GA WU438 UT WOS:A1996WU43800014 PM 9127890 ER PT J AU Persico, AM Uhl, GR AF Persico, AM Uhl, GR TI Transcription factors: Potential roles in drug-induced neuroplasticity SO REVIEWS IN THE NEUROSCIENCES LA English DT Review DE addiction; c-fos; craving; immediate-early gene; sensitization; tolerance; transcription factor; withdrawal ID IMMEDIATE-EARLY GENE; C-FOS EXPRESSION; MESSENGER-RNA EXPRESSION; TYROSINE-HYDROXYLASE GENE; ELEMENT-BINDING PROTEIN; CAMP-RESPONSE-ELEMENT; PRECIPITATED MORPHINE-WITHDRAWAL; ADRENAL-MEDULLARY CELLS; LONG-TERM FACILITATION; DOPAMINE-DEPLETED RATS AB Transcription factors act to regulate gene expression, Many transcription factor families have been discovered based on their roles in cell cycle events involved in development and oncogenesis, In post-mitotic neuronal cells, however, many transcription factor genes are ''transsynaptically'' regulated: their patterns of expression can be dramatically altered by extracellular stimuli, Transcription factor proteins can then potently influence expression of other genes, whose products can directly alter neuronal function. The central nervous system (CNS) displays varying degrees of neuroplasticity in adult life. Flexible neurochemical pathways that link extracellular stimuli to long-term modifications in neuronal functions are likely to contribute substantially to this neuroplasticity, This review summarizes evidence supporting central roles for transcription factors in such neurochemical cascades, It furthermore illustrates how drugs of abuse can trigger and modulate neuroadaptive processes that could conceivably contribute to clinically relevant addiction phenomena such as craving, tolerance, sensitization, and withdrawal. C1 NIDA,ADDICT RES CTR,OFF DIRECTOR,INTRAMURAL RES PROGRAM,NIH,BALTIMORE,MD 21224. JOHNS HOPKINS UNIV,SCH MED,DEPT NEUROL,BALTIMORE,MD 21205. JOHNS HOPKINS UNIV,SCH MED,DEPT NEUROSCI,BALTIMORE,MD 21205. LIBERO IST UNIV ROME,NEUROSCI LAB,ROME,ITALY. NR 330 TC 9 Z9 9 U1 1 U2 2 PU FREUND PUBLISHING HOUSE PI LONDON PA STE 500, CHESHAM HOUSE, 150 REGENT ST, LONDON, ENGLAND W1R 5FA SN 0334-1763 J9 REV NEUROSCIENCE JI Rev. Neurosci. PD OCT-DEC PY 1996 VL 7 IS 4 BP 233 EP 275 PG 43 WC Neurosciences SC Neurosciences & Neurology GA WH944 UT WOS:A1996WH94400001 PM 9044501 ER PT J AU Bassuk, EL Browne, A Buckner, JC AF Bassuk, EL Browne, A Buckner, JC TI Single mothers and welfare SO SCIENTIFIC AMERICAN LA English DT Article C1 CAMBRIDGE HOSP,CAMBRIDGE,MA 02139. HARVARD UNIV,SCH MED,CAMBRIDGE,MA 02138. NIMH,BETHESDA,MD 20892. RP Bassuk, EL (reprint author), BETTER HOMES FUND,NEWTON,MA, USA. NR 3 TC 38 Z9 38 U1 0 U2 0 PU SCI AMERICAN INC PI NEW YORK PA 415 MADISON AVE, NEW YORK, NY 10017 SN 0036-8733 J9 SCI AM JI Sci.Am. PD OCT PY 1996 VL 275 IS 4 BP 60 EP & PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VG523 UT WOS:A1996VG52300025 PM 8797298 ER PT J AU Badolato, R Oppenheim, JJ AF Badolato, R Oppenheim, JJ TI Role of cytokines, acute-phase proteins, and chemokines in the progression of rheumatoid arthritis SO SEMINARS IN ARTHRITIS AND RHEUMATISM LA English DT Review DE cytokines; acute phase proteins; chemokines; inflammation ID TUMOR-NECROSIS-FACTOR; C-REACTIVE PROTEIN; SERUM AMYLOID-A; CARTILAGE-PANNUS JUNCTION; INTERLEUKIN-1 RECEPTOR ANTAGONIST; HUMAN SYNOVIAL-CELLS; COLLAGEN-INDUCED ARTHRITIS; COLONY-STIMULATING FACTOR; MESSENGER-RNA EXPRESSION; GROWTH-FACTOR-BETA AB Rheumatoid arthritis (RA) has no firm etiologic basis. It progresses as an autoimmune disease and evolves into a chronic inflammatory joint disease complicated by recurrent episodes of systemic acute-phase reactions, which sometimes result in amyloidosis. Cytokines play a pivotol role in inflammation and the immune response, Proinflammatory cytokines such as interleukin-1, tumor necrosis factor alpha (TNF-alpha), and interleukin-6 are present at high levels in arthritic joints, and their blood concentration correlates with the severity of the RA. Some of the activities of the proinflammatory cytokines, such as stimulation of leukocyte infiltration and release of their proteolytic enzymes, may be mediated by acute phase proteins (APPs), such as C-reactive protein and serum amyloid A, and by chemokines such as interleukin-8. Cytokines, chemokines, and APPs reciprocally regulate each others' expression and activities, constituting a communication network between fibroblasts, macrophages, lymphocytes, and hepatocytes. Activation of the network results in inflammation and the progressive destruction of joints and systemic symptoms characteristic of RA. C1 NCI,FREDERICK CANC RES & DEV CTR,MOL IMMUNOREGULAT LAB,FREDERICK,MD 21702. RI Badolato, Raffaele/A-8081-2010 OI Badolato, Raffaele/0000-0001-7375-5410 NR 143 TC 124 Z9 129 U1 1 U2 3 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0049-0172 J9 SEMIN ARTHRITIS RHEU JI Semin. Arthritis Rheum. PD OCT PY 1996 VL 26 IS 2 BP 526 EP 538 DI 10.1016/S0049-0172(96)80041-2 PG 13 WC Rheumatology SC Rheumatology GA VQ582 UT WOS:A1996VQ58200003 PM 8916297 ER PT J AU Gutkind, JS VitaleCross, L AF Gutkind, JS VitaleCross, L TI The pathway linking small GTP-binding proteins of the Rho family to cytoskeletal components and novel signaling kinase cascades SO SEMINARS IN CELL & DEVELOPMENTAL BIOLOGY LA English DT Review DE actin; cytoskeleton; GTP-binding proteins; kinases; oncogenes; Ras; Rho ID NUCLEOTIDE EXCHANGE; RAS TRANSFORMATION; RAC1; ACTIVATION; CDC42; PROFILIN; ONCOGENE; GTPASES; DOMAIN; CELLS AB The Ras superfamily of GTPases comprises more than 50 members. Whereas Ras is known to play a central role in cell proliferation the Rho-family of Ras-related small GTP-binding proteins have been shown to regulate several aspects of cytoskeleton functioning. Recently, we and others have shown that members of the Rho-family of GTPases also control signaling cascades communicating the membrane to the nucleus, and that these GTP-binding proteins are integral components of growth-regulatory pathways. In this article, we will focus on the recent identification of molecules linking the Rho-family of GTPases to the cytoskeleton and to signaling kinase cascades. (C) 1996 Academic Press Ltd RP Gutkind, JS (reprint author), NIDR,MOL SIGNALLING UNIT,CELLULAR DEV & ONCOL LAB,NIH,9000 ROCKVILLE PIKE,BLDG 30,ROOM 212,BETHESDA,MD 20892, USA. RI Gutkind, J. Silvio/A-1053-2009 NR 41 TC 15 Z9 15 U1 0 U2 0 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON, ENGLAND NW1 7DX SN 1084-9521 J9 SEMIN CELL DEV BIOL JI Semin. Cell Dev. Biol. PD OCT PY 1996 VL 7 IS 5 BP 683 EP 690 DI 10.1006/scdb.1996.0084 PG 8 WC Cell Biology; Developmental Biology SC Cell Biology; Developmental Biology GA VY082 UT WOS:A1996VY08200009 ER PT J AU Klein, HG AF Klein, HG TI Transfusion medicine - Introduction SO SEMINARS IN HEMATOLOGY LA English DT Editorial Material RP Klein, HG (reprint author), NIH,WARREN G MAGNUSON CLIN CTR,DEPT TRANSFUS MED,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 SN 0037-1963 J9 SEMIN HEMATOL JI Semin. Hematol. PD OCT PY 1996 VL 33 IS 4 BP 275 EP 276 PG 2 WC Hematology SC Hematology GA VQ194 UT WOS:A1996VQ19400001 PM 8916301 ER PT J AU Leitman, SF Read, EJ AF Leitman, SF Read, EJ TI Hematopoietic progenitor cells SO SEMINARS IN HEMATOLOGY LA English DT Review ID COLONY-STIMULATING FACTOR; UMBILICAL-CORD-BLOOD; BONE-MARROW TRANSPLANTATION; HIGH-DOSE CHEMOTHERAPY; NON-HODGKINS-LYMPHOMA; LONG-TERM CULTURE; HUMAN-FETAL LIVER; PERIPHERAL-BLOOD; STEM-CELLS; AUTOLOGOUS TRANSPLANTATION RP Leitman, SF (reprint author), NIH,CC DTM,9000 ROCKVILLE PIKE,BETHESDA,MD 20892, USA. NR 128 TC 11 Z9 13 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0037-1963 J9 SEMIN HEMATOL JI Semin. Hematol. PD OCT PY 1996 VL 33 IS 4 BP 341 EP 358 PG 18 WC Hematology SC Hematology GA VQ194 UT WOS:A1996VQ19400007 PM 8916307 ER PT J AU Klein, HG Strauss, RG Schiffer, CA AF Klein, HG Strauss, RG Schiffer, CA TI Granulocyte transfusion therapy SO SEMINARS IN HEMATOLOGY LA English DT Article ID CHRONIC GRANULOMATOUS-DISEASE; AMPHOTERICIN-B; INDIUM-111-LABELED GRANULOCYTES; FILTRATION LEUKAPHERESIS; LEUKOCYTE TRANSFUSIONS; MYELOGENOUS LEUKEMIA; NEUTROPENIC PATIENTS; CONTROLLED TRIAL; FATE INVIVO; EFFICACY RP Klein, HG (reprint author), NIH,CTR CLIN,DEPT TRANSFUS MED,BLDG 10,RM 1C71,9000 ROCKVILLE PIKE,BETHESDA,MD 20892, USA. NR 63 TC 22 Z9 22 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0037-1963 J9 SEMIN HEMATOL JI Semin. Hematol. PD OCT PY 1996 VL 33 IS 4 BP 359 EP 368 PG 10 WC Hematology SC Hematology GA VQ194 UT WOS:A1996VQ19400008 PM 8916308 ER PT J AU Kubbutat, MHG Vousden, KH AF Kubbutat, MHG Vousden, KH TI Role of E6 and E7 oncoproteins in HPV-induced anogenital malignancies SO SEMINARS IN VIROLOGY LA English DT Review DE E6; E7; HPV-induced malignancies; viral oncoproteins ID HUMAN PAPILLOMAVIRUS TYPE-16; RETINOBLASTOMA GENE-PRODUCT; CARCINOMA CELL-LINES; UBIQUITIN-CONJUGATING ENZYME; CALCIUM-BINDING PROTEIN; CYCLIN-DEPENDENT KINASE; P53 DNA-BINDING; MUTATIONAL ANALYSIS; TRANSCRIPTIONAL ACTIVATION; HUMAN KERATINOCYTES AB The association between infection with high-risk human papilloma virus (HPV) and cervical cancer is now well established and the viral proteins E6 and E7 have been identified as oncoproteins. The mechanisms by which E6 and E7 overcome normal cell cycle control involve the inactivation of two major tumor suppressor proteins, namely p53 and the retinoblastoma gene product (pRB). The unscheduled entry into S-phase enforced by E7 through interaction with pRB in combination with the disruption of the p53 checkpoint by EG can lead to genomic instability that might represent a basic step towards the development of HIV-induced anogenital malignancies. RP Kubbutat, MHG (reprint author), NCI,FREDERICK CANC RES & DEV CTR,ABL BASIC RES PROGRAM,FREDERICK,MD 21701, USA. NR 113 TC 26 Z9 27 U1 1 U2 4 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON, ENGLAND NW1 7DX SN 1044-5773 J9 SEMIN VIROL JI Semin. Virol. PD OCT PY 1996 VL 7 IS 5 BP 295 EP 304 DI 10.1006/smvy.1996.0037 PG 10 WC Virology SC Virology GA WE092 UT WOS:A1996WE09200002 ER PT J AU Eastman, HB Fawcett, TW Udelsman, R Holbrook, NJ AF Eastman, HB Fawcett, TW Udelsman, R Holbrook, NJ TI Effects of perturbations of the hypothalamic-pituitary-adrenal axis on the acute phase response: Altered C/EBP and acute phase response gene expression in lipopolysaccharide-treated rats SO SHOCK LA English DT Article ID ALPHA-1-ACID GLYCOPROTEIN GENE; NF-KAPPA-B; BINDING-PROTEIN ISOFORMS; TRANSCRIPTION FACTORS; GLUCOCORTICOID RECEPTOR; NUCLEAR-PROTEIN; HEPATOMA-CELLS; INTERLEUKIN-6; LIVER; ELEMENT AB In this study, we investigated the influence of long term perturbations of the hypothalamic-pituitary-adrenal axis on the acute phase response elicited following lipopolysaccharide (LPS) challenge in rats. Specifically, we examined the effects of either long term absence of glucocorticoids (adrenalectomized rats treated with placebo chronic release pellets) or extended exposure to pharmacologic levels of glucocorticoids (adrenalectomized rats treated with dexamethasone chronic release pellets) on the expression of selected acute phase proteins and various members of the CCAAT/enhancer-binding protein (C/EBP) family of transcription factors. Both hypothalamic-pituitary-adrenal axis manipulations resulted in a reduction of the acute phase response as assessed by the LPS-mediated induction of acute phase proteins and C/EBP gene expression, with dexamethasone treatment exhibiting a greater inhibitory effect than adrenalectomy. Induction of hemopexin, alpha(1)-acid glycoprotein, alpha(2)-macroglobulin, GADD153, C/EBP beta, and C/EBP delta mRNAs by LPS were all abolished in dexamethasone-treated rats. These findings have direct implications for patients undergoing chronic high dose glucocorticoid therapy. C1 NIA, GERONTOL RES CTR, GENE EXPRESS & AGING SECT, BALTIMORE, MD 21224 USA. JOHNS HOPKINS UNIV HOSP, LAB ENDOCRINE SURG, BALTIMORE, MD 21287 USA. NR 40 TC 19 Z9 19 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1073-2322 J9 SHOCK JI Shock PD OCT PY 1996 VL 6 IS 4 BP 286 EP 292 DI 10.1097/00024382-199610000-00011 PG 7 WC Critical Care Medicine; Hematology; Surgery; Peripheral Vascular Disease SC General & Internal Medicine; Hematology; Surgery; Cardiovascular System & Cardiology GA VN106 UT WOS:A1996VN10600011 PM 8902947 ER PT J AU DAgostino, RB Burke, G OLeary, D Rewers, M Selby, J Savage, PJ Saad, MF Bergman, RN Howard, G Wagenknecht, L Haffner, SM AF DAgostino, RB Burke, G OLeary, D Rewers, M Selby, J Savage, PJ Saad, MF Bergman, RN Howard, G Wagenknecht, L Haffner, SM TI Ethnic differences in carotid wall thickness - The Insulin Resistance Atherosclerosis Study SO STROKE LA English DT Article DE atherosclerosis; blacks; ethnic groups; Hispanic Americans; insulin ID DEPENDENT DIABETES-MELLITUS; CARDIOVASCULAR RISK-FACTORS; NON-HISPANIC WHITES; BIETHNIC COLORADO POPULATION; IMPAIRED GLUCOSE-TOLERANCE; ISCHEMIC HEART-DISEASE; INTIMA-MEDIA THICKNESS; MIDDLE-AGED ADULTS; B-MODE ULTRASOUND; MEXICAN-AMERICANS AB Background and Purpose Ethnic differences in cardiovascular disease (CVD) morbidity and mortality have been observed in US adults. However, little data exist on differences in indices of preclinical atherosclerosis such as carotid wall intima-media thickness (IMT) for US non-Hispanic whites, Hispanics, and blacks. This study was undertaken to determine whether there were ethnic differences in carotid wall IMT. Methods Internal carotid artery (ICA) IMT and common carotid artery (CCA) IMT, indices of atherosclerosis, were assessed with the use of B-mode ultrasound in 1020 nondiabetic participants in the Insulin Resistance Atherosclerosis Study, a multicenter study designed to examine the association between insulin resistance and carotid atherosclerosis. The study included 281 blacks, 329 Hispanics, and 410 non-Hispanic whites aged 40 to 69 years. Results Blacks had significantly greater CCA IMT than non-Hispanic whites (865 versus 808 mu m); this remained significant after adjustment for major CVD risk factors and insulin sensitivity (864 versus 823 mu m). There were no significant differences in ICA IMT between blacks and non-Hispanic whites. Hispanics had significantly lesser CCA IMT than non-Hispanic whites (749 versus 776 mu m), and these differences remained significant after adjustment for traditional cardiovascular risk factors and insulin sensitivity (750 versus 778 mu m). There were no significant differences in ICA IMT between non-Hispanic whites and Hispanics. Conclusions We conclude that ethnic differences exist in CCA but not in ICA IMT in nondiabetic subjects. These differences in IMT, which are indicators of atherosclerosis, are a noninvasive measure that is consistent with some of the data on clinical end points. These differences may be associated with the observed differences in CVD morbidity and mortality among major ethnic groups in the United States. C1 TUFTS UNIV NEW ENGLAND MED CTR,DEPT RADIOL,BOSTON,MA 02111. UNIV COLORADO,SCH MED,DEPT PREVENT MED BIOMETR,DENVER,CO. KAISER PERMANENTE,DIV RES,OAKLAND,CA. NHLBI,DIV EPIDEMIOL & CLIN APPLICAT,NIH,BETHESDA,MD 20892. UNIV SO CALIF,SCH MED,DEPT MED,LOS ANGELES,CA 90033. UNIV SO CALIF,SCH MED,DEPT PHYSIOL & BIOPHYS,LOS ANGELES,CA 90033. UNIV TEXAS,HLTH SCI CTR,DEPT MED,SAN ANTONIO,TX 78284. RP DAgostino, RB (reprint author), BOWMAN GRAY SCH MED,DEPT PUBL HLTH SCI,MED CTR BLVD,WINSTON SALEM,NC 27157, USA. RI Dagostino Jr, Ralph/C-4060-2017 OI Dagostino Jr, Ralph/0000-0002-3550-8395 FU NHLBI NIH HHS [HL-47887, HL-47889, HL-47890] NR 54 TC 110 Z9 111 U1 0 U2 1 PU AMER HEART ASSOC PI DALLAS PA 7272 GREENVILLE AVENUE, DALLAS, TX 75231-4596 SN 0039-2499 J9 STROKE JI Stroke PD OCT PY 1996 VL 27 IS 10 BP 1744 EP 1749 PG 6 WC Clinical Neurology; Peripheral Vascular Disease SC Neurosciences & Neurology; Cardiovascular System & Cardiology GA VK607 UT WOS:A1996VK60700007 PM 8841322 ER PT J AU Lin, HJ Wolf, PA KellyHayes, M Belser, AS Kase, CS Benjamin, EJ DAgostino, RB AF Lin, HJ Wolf, PA KellyHayes, M Belser, AS Kase, CS Benjamin, EJ DAgostino, RB TI Stroke severity in atrial fibrillation - The Framingham study SO STROKE LA English DT Article DE atrial fibrillation; stroke outcome; risk factors; mortality; disability evaluation ID CEREBRAL BLOOD-FLOW; PROGNOSTIC FACTORS; COMMUNITY-STROKE; RISK FACTOR; FOLLOW-UP; SURVIVAL; MORTALITY AB Background and Purpose Stroke occurring with atrial fibrillation (AF) is more likely to be fatal or more severe than non-AF stroke based on clinical series, but data from prospective epidemiological studies are sparse and inconsistent. Methods Over 40-year follow-up of the original 5070 Framingham cohort, 501 initial ischemic strokes, including 103 with AF, were analyzed. Stroke severity was rated as none, mild, moderate, severe, or fatal. Since 1981, functional status indicated by the Barthel index has been evaluated acutely and at 3, 6, and 12 months. Severity and functional status of AF strokes were compared with non-AF strokes using chi(2) test and Student's t test. Thirty-day mortality was assessed by logistic regression analyses. Results AF was associated with increased stroke severity (P=.048). Thirty-day mortality was greater in AF strokes than in non-AF strokes (25% versus 14%). The multivariate-adjusted odds ratio for 30-day mortality for AF subjects was 1.84 (95% confidence interval, 1.04 to 3.27). Since 1981, follow-up was available for 150 initial ischemic strokes, including 30 with AF. Compared with the non-AF group, the AF group had poorer survival and more recurrences during 1 year of follow-up. The AF subjects had lower mean Barthel index scores acutely (29.6 versus 58.6, P<.001) and at 3 months (P=.005), 6 months (P=.003), and 12 months (P=.130) after stroke among survivors. Conclusions Ischemic stroke associated with AF was nearly twice as likely to be fatal as non-AF stroke. Recurrence was more frequent, and functional deficits were more likely to be severe among survivors. Since stroke is usually the initial manifestation of embolism in AF, prevention is critical to reducing disability and mortality. C1 BOSTON UNIV,SCH MED,DEPT NEUROL,BOSTON,MA 02118. BOSTON UNIV,SCH PUBL HLTH,DEPT BIOSTAT & EPIDEMIOL,BOSTON,MA. BOSTON UNIV,SCH MED,DEPT CARDIOL,BOSTON,MA 02118. BOSTON UNIV,DEPT MATH,BOSTON,MA 02118. NHLBI,FRAMINGHAM STUDY,FRAMINGHAM,MA. FU NHLBI NIH HHS [N01-HC-38038]; NINDS NIH HHS [2 RO1-NS-17950-15] NR 24 TC 571 Z9 583 U1 1 U2 14 PU AMER HEART ASSOC PI DALLAS PA 7272 GREENVILLE AVENUE, DALLAS, TX 75231-4596 SN 0039-2499 J9 STROKE JI Stroke PD OCT PY 1996 VL 27 IS 10 BP 1760 EP 1764 PG 5 WC Clinical Neurology; Peripheral Vascular Disease SC Neurosciences & Neurology; Cardiovascular System & Cardiology GA VK607 UT WOS:A1996VK60700010 PM 8841325 ER PT J AU Temeck, BK Koong, SS Reynolds, JC Pass, HI AF Temeck, BK Koong, SS Reynolds, JC Pass, HI TI Somatostatin analogue in the localization and treatment of bronchial carcinoid tumours SO SURGICAL ONCOLOGY-OXFORD LA English DT Review DE somatostatin; carcinoid; ectopic ACTH; receptor; analogues ID CELL LUNG-CANCER; RECEPTOR SCINTIGRAPHY; NEUROENDOCRINE TUMORS; IN-111 PENTETREOTIDE; CUSHINGS-SYNDROME; INVITRO; INVIVO AB Octreotide scanning is increasingly being used to detect tumours with somatostatin receptors. Moreover, there is growing interest in the use of somatostatin analogues for the treatment of tumours with somatostatin receptors. This review documents the use at our institution of the octreotide scan in three patients with intrathoracic pathology, and comments on overall experience in the literature with this technology. C1 NCI,HEAD THORAC ONCOL SECT,SURG BRANCH,NIH,BETHESDA,MD 20892. NR 27 TC 1 Z9 1 U1 0 U2 0 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, OXON, ENGLAND OX5 1GB SN 0960-7404 J9 SURG ONCOL JI Surg. Oncol.-Oxf. PD OCT-DEC PY 1996 VL 5 IS 5-6 BP 195 EP 200 PG 6 WC Oncology; Surgery SC Oncology; Surgery GA WT903 UT WOS:A1996WT90300001 PM 9129131 ER PT J AU Liu, LX Monsma, FJ Sibley, DR Chiodo, LA AF Liu, LX Monsma, FJ Sibley, DR Chiodo, LA TI D-2L, D-2S, and D-3 dopamine receptors stably transfected into NG108-15 cells couple to a voltage-dependent potassium current via distinct G protein mechanisms SO SYNAPSE LA English DT Article DE transfection; DA receptors; D-2 receptor isoforms; D-2 receptors; G(O alpha); G(S alpha); G proteins; whole-cell patch clamp; signal transduction ID GTP-BINDING PROTEIN; HAMSTER OVARY CELLS; ALPHA-SUBUNIT; FUNCTIONAL EXPRESSION; PITUITARY-CELLS; CHANNELS; D3; CLONING; FORMS; K+ AB The D-2-like dopamine (DA) receptor family has continued to expand and now includes the D-2-short (D-2S) and DB-long (D-2L) receptor isoforms and the D-3 and D-4 receptors. The D-2 receptor isoforms differ in length by 29 amino acids within the third cytoplasmic loop, a region of the receptor believed to be important for G protein coupling. This observation has led to the hypothesis that the two isoforms of the D-2 receptor may utilize different signal transduction pathways when present in the same cell. The D-2 and D-3 receptors, although mostly different, show some common amino acid sequences within the third cytoplasmic loop. Thus, it is possible that the D-2 and D-3 receptors may employ similar signal transduction pathways. To test these hypotheses directly, NG108-15 neuroblastoma-glioma hybrid cells were stably transfected to express either the Das, D-2L, Or D-3 DA receptors. All transfected but not untransfected NG108-15 cells demonstrated a dose-dependent reduction in the peak whole-cell potassium (K+) current in response to receptor activation by DA or the DA receptor agonists quinpirole (QUIN) and apomorphine (APO). The modulation of K+ current by D-2S receptor stimulation was prevented by pretreatment of the cells with cholera toxin (20 mu g/ml for 18 h), whereas pertussis toxin pretreatment (500 ng/ml for 4 h) completely blocked the effects of D-2L and D-3 receptor activation. These observations suggest that the signal transduction mechanisms involved in coupling the two isoforms of the D-2 receptor to the K+ current are different, whereas the D-2L and D-3 receptor coupling mechanisms may be similar. In direct support of this hypothesis, it was observed that the intracellular application of a polyclonal antibody that is specific for the G(O alpha) subunit completely blocked the ability of D-2L and D-3 receptors to modulate outward K+ currents. In contrast, the D-2S-mediated modulation of K+ currents was blocked by intracellular application of an antibody recognizing G(S alpha) but not G(O alpha). These findings demonstrate that D-2S and D-2L receptors are able to couple to a common effector in a cell via two G protein pathways. (C) 1996 Wiley-Liss, Inc. C1 TEXAS TECH UNIV,HLTH SCI CTR,DEPT PHARMACOL,SCH MED,LUBBOCK,TX 79430. NINCDS,MOL NEUROPHARMACOL SECT,EXPT THERAPEUT BRANCH,NIH,BETHESDA,MD 20892. FU NIMH NIH HHS [MH-41557] NR 35 TC 35 Z9 35 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC 605 THIRD AVE, NEW YORK, NY 10158-0012 SN 0887-4476 J9 SYNAPSE JI Synapse PD OCT PY 1996 VL 24 IS 2 BP 156 EP 164 PG 9 WC Neurosciences SC Neurosciences & Neurology GA VK329 UT WOS:A1996VK32900007 PM 8890457 ER PT J AU Heidbreder, CA Shippenberg, TS AF Heidbreder, CA Shippenberg, TS TI Evidence for an involvement of muscarinic cholinergic systems in the induction but not expression of behavioral sensitization to cocaine SO SYNAPSE LA English DT Article DE acetylcholine; cocaine; sensitization; scopolamine; locomotor activity; stereotypy ID VENTRAL TEGMENTAL AREA; REWARDING BRAIN-STIMULATION; RAT HIPPOCAMPAL SLICES; FREELY MOVING RATS; NUCLEUS-ACCUMBENS; EXTRACELLULAR DOPAMINE; SUBSTANTIA-NIGRA; AUTOMATED MEASUREMENT; INVIVO MICRODIALYSIS; PSYCHOMOTOR STIMULANT AB The present study was designed to determine whether the muscarinic cholinergic antagonist scopolamine can prevent the expression and induction of sensitization to the locomotor-activating effects of cocaine. Rats received one daily injection of cocaine (20 mg/kg i.p.) for 5 days. Two days after withdrawal of pretreatment, rats were pretreated with scopolamine (3.0 mg/kg s.c) or its vehicle and challenged 15 min later with either saline or cocaine (20-30 mg/kg i.p.). In a second set of experiments, scopolamine (3.0 mg/kg s.c) or its vehicle was given in combination with either saline or cocaine (20 mg/kg i.p.) for 5 days. Activity in response to saline and to cocaine (20 mg/kg i.p.) was assessed on day 7. The effects of acute administration of scopolamine (3.0 mg/kg s.c.) on cocaine-induced locomotor activity were also assessed. Acute administration of scopolamine increased both distance traveled and time spent in stereotypy. When scopolamine was administered 15 min prior to an acute injection of cocaine, a significant increase in the behavioral response to cocaine was seen. Daily injections of cocaine for 5 days produced sensitized behavioral responses to a subsequent cocaine challenge. Acute administration of scopolamine to animals preexposed and sensitized to cocaine did not disrupt the expression of sensitization to the locomotor and stereotypic effects of cocaine. In contrast, when scopolamine was given in combination with cocaine for 5 days, sensitization to the locomotor-activating effects of cocaine was prevented. These results suggest an important role of cholinergic muscarinic systems in mediating sensitization to the locomotor-activating effects of cocaine, which occurred after the repeated context-independent administration of this agent. In contrast, the enhanced stereotypic effects in response to the repeated administration of cocaine seem to be independent of alterations in muscarinic cholinergic transmission. (C) 1996 Wiley-Liss, Inc. C1 NIDA,NIH,PRECLIN PHARMACOL LAB,BEHAV PHARMACOL & GENET SECT,BALTIMORE,MD 21224. NR 85 TC 35 Z9 37 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC 605 THIRD AVE, NEW YORK, NY 10158-0012 SN 0887-4476 J9 SYNAPSE JI Synapse PD OCT PY 1996 VL 24 IS 2 BP 182 EP 192 DI 10.1002/(SICI)1098-2396(199610)24:2<182::AID-SYN10>3.3.CO;2-H PG 11 WC Neurosciences SC Neurosciences & Neurology GA VK329 UT WOS:A1996VK32900010 PM 8890460 ER PT J AU Oravecz, T Pall, M Norcross, MA AF Oravecz, T Pall, M Norcross, MA TI Effect of the N-L, T-cell and myeloid section antibodies on homotypic cell adhesion and HIV-1 induced syncytium formation. SO TISSUE ANTIGENS LA English DT Meeting Abstract C1 NIH,FDA,CBER,BETHESDA,MD 20892. NR 0 TC 0 Z9 0 U1 0 U2 0 PU MUNKSGAARD INT PUBL LTD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0001-2815 J9 TISSUE ANTIGENS JI Tissue Antigens PD OCT PY 1996 VL 48 IS 4-II BP NL406 EP NL406 PG 1 WC Cell Biology; Immunology; Pathology SC Cell Biology; Immunology; Pathology GA VR930 UT WOS:A1996VR93000364 ER PT J AU Drahushuk, AT McGarrigle, BP Tai, HL Kitareewan, S Goldstein, JA Olson, JR AF Drahushuk, AT McGarrigle, BP Tai, HL Kitareewan, S Goldstein, JA Olson, JR TI Validation of precision-cut liver slices in dynamic organ culture as an in vitro model for studying CYP1A1 and CYP1A2 induction SO TOXICOLOGY AND APPLIED PHARMACOLOGY LA English DT Article ID ADULT-RAT HEPATOCYTES; DIBENZO-P-DIOXINS; MICROSOMAL CYTOCHROME-P-450; MOUSE HEPATOCYTES; HEPATIC-UPTAKE; AH RECEPTOR; CELL-LINE; METABOLISM; 2,3,7,8-TETRACHLORODIBENZO-PARA-DIOXIN; BINDING AB The utilization of precision-cut liver slices in dynamic organ culture as an in vitro model was validated by comparing the induction of the biomarker responses following in vitro (rat liver slice) and in vivo exposure of rats to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The biomarker responses investigated were cytochrome P450s 1A1 and 1A2 (CYP1A1 and CYP1A2) mRNA, protein, and activities. Precision-cut rat liver slices were incubated in dynamic organ culture for 24 hr with medium containing 0.001-10 nM TCDD or medium without TCDD (control). The resultant mean TCDD concentration in the slices ranged from 19 to 80,925 ppt (wet wt), respectively. A concentration-dependent induction of CYP1A1 mRNA, protein, and activities and a more modest induction of CYP1A2 mRNA was observed in liver slices at all medium concentrations of TCDD. The O-demethylation of 7-methoxyresorufin, a marker for CYP1A2 activity, was induced at TCDD medium levels of 0.01 nM and greater, whereas a detectable increase in CYP1A2 protein occurred only at the higher concentrations. Comparable liver concentrations of TCDD (8-64,698 ppt wet wt) were achieved at 24 hr following a single in vivo exposure of rats to TCDD at doses ranging from 0.002 to 5 mu g/kg po. Concentration-effect and dose-response relationships for induction of CYP1A1 and CYP1A2 were similar following in vitro and in vivo exposure to TCDD, although the magnitude of induction was greater for in vivo exposure. The data support the use of liver slices in dynamic organ culture for assessing the relative in vivo potency of a compound to induce CYP1A1 and CYP1A2. Human tissue can also be readily utilized in this in vitro model to predict the biological and toxicological effects of a given in vivo exposure to TCDD. (C) 1996 academic Press, Inc. C1 NIEHS,RES TRIANGLE PK,NC 27709. RP Drahushuk, AT (reprint author), SUNY BUFFALO,DEPT PHARMACOL & TOXICOL,BUFFALO,NY 14214, USA. RI Goldstein, Joyce/A-6681-2012 FU NIEHS NIH HHS [ES06556-01] NR 47 TC 56 Z9 57 U1 0 U2 0 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0041-008X J9 TOXICOL APPL PHARM JI Toxicol. Appl. Pharmacol. PD OCT PY 1996 VL 140 IS 2 BP 393 EP 403 DI 10.1006/taap.1996.0236 PG 11 WC Pharmacology & Pharmacy; Toxicology SC Pharmacology & Pharmacy; Toxicology GA VN707 UT WOS:A1996VN70700023 PM 8887457 ER PT J AU McGregor, DB Riach, C Cattanach, P Edwards, I Shepherd, W Caspary, WJ AF McGregor, DB Riach, C Cattanach, P Edwards, I Shepherd, W Caspary, WJ TI Mutagenic responses of L5178Y mouse cells at the tk and hprt loci SO TOXICOLOGY IN VITRO LA English DT Article ID HAMSTER OVARY CELLS; SPECIES-SPECIFIC DIFFERENCES; THYMIDINE KINASE LOCUS; LYMPHOMA-CELLS; / MUTANTS; TRIFLUOROTHYMIDINE-RESISTANT; ETHYL METHANESULFONATE; CHROMOSOME ANALYSIS; MAMMALIAN-CELLS; ASSAY SYSTEM AB The expression of multiple recessive genes by aberrant mitotic lesions plays a major part in carcinogenesis. These lesions include intragenic mutations as well as chromosomal lesions. An in vitro model for studying carcinogenesis should respond to all these lesions. Mutagenesis studies that target hemizygous loci may not be useful in studying chromosomal mechanisms because lesions incorporating essential genes already missing on the inactive, homologous chromosome may be lethal to the cell. Cells heterozygous at the selectable gene may survive. Using L5178Y mouse cells, we compared the mutagenic responses at the heterozygous rk and hemizygous hprt loci to four chemicals-benzidine dihydrochloride, diglycidylresorcinol ether, nitrofen and p-benzoquinone dioxime. None of the compounds induced clear positive responses at the hprt locus. In contrast, all the compounds induced clear or marginal mutagenic responses at the tk locus. These data are consistent with the expectation that heterozygous loci can detect lesions that are refractory to hemizygous loci. Published by Elsevier Science Ltd. C1 INVERESK RES INT LTD,MUSSELBURGH EH21 7UB,SCOTLAND. NIH,CANC GENET GRP,RES TRIANGLE PK,NC 27709. NR 37 TC 3 Z9 3 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, ENGLAND OX5 1GB SN 0887-2333 J9 TOXICOL IN VITRO JI Toxicol. Vitro PD OCT PY 1996 VL 10 IS 5 BP 643 EP 647 DI 10.1016/S0887-2333(96)90030-2 PG 5 WC Toxicology SC Toxicology GA VY583 UT WOS:A1996VY58300016 PM 20650247 ER PT J AU Jefferson, WN Teng, C Newbold, RR AF Jefferson, WN Teng, C Newbold, RR TI Methodologies for isolating estrogen-responsive proteins as markers of environmental toxicants SO TOXICOLOGY METHODS LA English DT Article DE complement C3; electroelution; estrogen; estrogenic markers; lactoferrin; protein purification; uterine luminal fluid; uterus ID HUMAN LACTOFERRIN; MOUSE; LACTOTRANSFERRIN; EXPRESSION; UTERUS; GROWTH; GENE AB New and refined methods for the purification of two major estrogen-responsive proteins in murine uterine luminal fluid (ULF) are described. Lactoferrin (LF; approx. 70 kDa), was purified using modifications of procedures previously reported. Using column chromatography and electroelution, the protein was purified to a single band demonstrated by gel electrophoresis. An antibody to the purified protein was then raised in rabbits. Rabbit serum was IgG purified and used in Western blotting and immunocytochemical techniques, The serum IgG purified LF antibody demonstrated only one band in Western blotting procedures using enhanced chemiluminescence (ECL) as a detection method. Another estrogen-inducible protein with an approximate molecular weight of 63 kDa in mouse ULF, purified to a single band by gel electrophoresis and electroelution, was determined to be complement C3 through sequence analysis. Similar to LF, C3 has been previously described as an estrogen-responsive protein in the rodent and human reproductive tract. C3 antibody was purchased commercially and was used for Western blotting and immunohistochemistry. Since interest is increasing in identifying biological markers of hormone-disrupting chemicals, the purification techniques described in this report can be useful in identifying proteins that are potential markers of estrogen action. Antibodies to these proteins can then be used as markers of environmental estrogenic toxicants. In fact, LF and C3 are currently being used to screen for sensitive markers of estrogenicity. Following the methods described in this report, other proteins can be identified and used to screen for possible effects of environmental agents. C1 NIEHS,DEV ENDOCRINOL SECT,REPROD TOXICOL GRP,TOXICOL LAB,ENVIRONM TOXICOL PROGRAM,NIH,RES TRIANGLE PK,NC 27709. NIEHS,GENE REGULAT GRP,REPROD & DEV TOXICOL LAB,ENVIRONM BIOL PROGRAM,DIV INTRAMURAL RES,NIH,RES TRIANGLE PK,NC. NR 17 TC 10 Z9 11 U1 1 U2 2 PU TAYLOR & FRANCIS PI BRISTOL PA 1900 FROST ROAD, SUITE 101, BRISTOL, PA 19007-1598 SN 1051-7235 J9 TOXICOL METHOD JI Toxicol. Method. PD OCT-DEC PY 1996 VL 6 IS 4 BP 183 EP 192 DI 10.3109/15376519609066119 PG 10 WC Toxicology SC Toxicology GA WP497 UT WOS:A1996WP49700001 ER PT J AU Chanturiya, AN Nikolaenko, AN Shatursky, OY Lishko, VK AF Chanturiya, AN Nikolaenko, AN Shatursky, OY Lishko, VK TI Probing the structure-function relationship of alpha-latrotoxin-formed channels with antibodies and pronase SO TOXICON LA English DT Article ID MEMBRANES; RECEPTOR AB The major toxic component of black widow spider (Latrodectus mactans tredecimguttatus) venom, alpha-latrotoxin, is known to form ionic channels in different membranes, In order to probe the extramembrane domains of alpha-latrotoxin molecule, alpha-latrotoxin channels in planar lipid membrane were treated with antibodies to latrotoxin or with pronase added to different sides of the membrane. It was found that antibody addition to the same side as the toxin (cis) decreased channel conductance only at positive potentials across the membrane. In contrast, trans side addition of antibodies changed the channel conductance at both positive and negative potentials: at positive potential conductance first slightly increased then decreased by more then 50%; at negative potential it decreased much more quickly, to only about 20% of the initial value. No dependence on membrane potential was found for pronase treatment of incorporated channels. For both cis and a trans application of pronase, channel selectivity for Ca2+, Mg2+, Ba2+ and K+, Na+, Li+ ions did not change significantly but Cd2+ block was decreased. Trans pronase treatment also resulted in some rectification of I/V curves and an increase in channel conductance. We interpret these findings as evidence that alpha-latrotoxin channel has protruding parts on both sides of the membrane and that its conformation in the membrane depends on membrane potential. Copyright (C) 1996 Elsevier Science Ltd C1 AV PALLADIN BIOCHEM INST,UA-252030 KIEV,UKRAINE. RP Chanturiya, AN (reprint author), NICHHD,THEORET & PHYS BIOL LAB,NIH,BD 10,RM 10D04,10 CTR DR,BETHESDA,MD 20892, USA. NR 23 TC 4 Z9 4 U1 0 U2 2 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, ENGLAND OX5 1GB SN 0041-0101 J9 TOXICON JI Toxicon PD OCT PY 1996 VL 34 IS 10 BP 1157 EP 1164 DI 10.1016/0041-0101(96)00053-0 PG 8 WC Pharmacology & Pharmacy; Toxicology SC Pharmacology & Pharmacy; Toxicology GA VV098 UT WOS:A1996VV09800008 PM 8931256 ER PT J AU Busch, MP Collier, A Gernsheimer, T Carrington, JD Flanigan, TP Kashkari, M Kennedy, M Kumar, PN Lane, TA Mellors, JW Mohandas, K Pollard, RB Viele, M Yomtovian, R Holland, PV McCurdy, PR AF Busch, MP Collier, A Gernsheimer, T Carrington, JD Flanigan, TP Kashkari, M Kennedy, M Kumar, PN Lane, TA Mellors, JW Mohandas, K Pollard, RB Viele, M Yomtovian, R Holland, PV McCurdy, PR TI The viral activation transfusion study (VATS): Rationale, objectives, and design overview SO TRANSFUSION LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; VERSUS-HOST DISEASE; POLYMERASE CHAIN-REACTION; CD4+ T-CELLS; BLOOD-TRANSFUSION; HIV-1 INFECTION; CYTOMEGALOVIRUS-INFECTION; TYPE-1 INFECTION; PLASMA; LYMPHOCYTES C1 IRWIN MEM BLOOD CTR,SCI SERV,SAN FRANCISCO,CA 94118. UNIV WASHINGTON,AIDS CLIN TRIAL UNIT,SEATTLE,WA 98195. UNIV WASHINGTON,DEPT MED HEMATOL,SEATTLE,WA 98195. PUGET SOUND BLOOD CTR,PLATELET IMMUNNOL LAB & MED TRANSFUS SERV,SEATTLE,WA 98104. NEW ENGLAND RES INST,WATERTOWN,MA 02172. MT SINAI MED CTR,BLOOD BANK,NEW YORK,NY 10029. OHIO STATE UNIV,TRANSFUS SERV LABS,COLUMBUS,OH 43210. GEORGETOWN UNIV,DIV INFECT DIS,WASHINGTON,DC. UNIV CALIF SAN DIEGO,SCH MED,DEPT PATHOL,SAN DIEGO,CA 92103. UNIV CALIF SAN DIEGO,MED CTR,TRASFUS SERV,SAN DIEGO,CA 92103. UNIV PITTSBURGH,PITT TREATMENT EVALUAT CTR,PITTSBURGH,PA. UNIV TEXAS,MED BRANCH,GALVESTON,TX 77550. UNIV CALIF SAN FRANCISCO,BLOOD BANK,SAN FRANCISCO,CA 94143. UNIV CALIF SAN FRANCISCO,BLOOD DONOR CTR,SAN FRANCISCO,CA 94143. UNIV HOSP CLEVELAND,BLOOD BANK TRUNSFUS MED SERV,CLEVELAND,OH 44106. SACRAMENTO MED FDN,CTR BLOOD,SACRAMENTO,CA. NHLBI,BLOOD RESOURCES PROGRAM,DIV BLOOD DIS & RESOURCES,BETHESDA,MD 20892. BROWN UNIV,MIRIAM HOSP,CTR IMMUNOL,PROVIDENCE,RI. BROWN UNIV,MIRIAM HOSP,DIV CLIN IMMUNOL & GEOG MED,PROVIDENCE,RI. UNIV N CAROLINA,CHAPEL HILL,NC. UNIV CALIF SAN FRANCISCO,MT ZION HOSP,SAN FRANCISCO,CA 94143. NEW ENGLAND RES CTR,WATERTOWN,MA. BETH ISRAEL HOSP,BOSTON,MA 02215. HARVARD UNIV,CAMBRIDGE,MA 02138. UNIV MINNESOTA,MINNEAPOLIS,MN 55455. RP Busch, MP (reprint author), IRWIN MEM BLOOD CTR,RES SERV,270 MASONIC AVE,SAN FRANCISCO,CA 94118, USA. NR 65 TC 38 Z9 38 U1 0 U2 0 PU AMER ASSOC BLOOD BANKS PI BETHESDA PA 8101 GLENBROOK RD, BETHESDA, MD 20814-2749 SN 0041-1132 J9 TRANSFUSION JI Transfusion PD OCT PY 1996 VL 36 IS 10 BP 854 EP 859 DI 10.1046/j.1537-2995.1996.361097017169.x PG 6 WC Hematology SC Hematology GA VM881 UT WOS:A1996VM88100002 PM 8863770 ER PT J AU Lenfant, C AF Lenfant, C TI Acknowledgment of research sponsors: It pays to advertise SO TRANSFUSION LA English DT Article AB The Committee [House Appropriations Subcommittee on Labor, Health and Human Services, and Education] believes it is critical for NIH [National Institutes of Health] to use all the media at its command to publicize the benefits and results of NM research, in order to solidify the general public support of biomedical research and to identify...the funding source for these breakthroughs in the public's mind. The Committee also urges...whatever steps are necessary to ensure that grantees acknowledge NIH's funding contribution when they publicize their research findings. RP Lenfant, C (reprint author), NHLBI,NIH,BLDG 31,ROOM 5A52,BETHESDA,MD 20892, USA. NR 1 TC 0 Z9 0 U1 1 U2 1 PU AMER ASSOC BLOOD BANKS PI BETHESDA PA 8101 GLENBROOK RD, BETHESDA, MD 20814-2749 SN 0041-1132 J9 TRANSFUSION JI Transfusion PD OCT PY 1996 VL 36 IS 10 BP 935 EP 936 DI 10.1046/j.1537-2995.1996.361097059065.x PG 2 WC Hematology SC Hematology GA VM881 UT WOS:A1996VM88100016 PM 8928220 ER PT J AU Hengen, PN AF Hengen, PN TI Purification of GST fusion proteins SO TRENDS IN BIOCHEMICAL SCIENCES LA English DT Article ID GLUTATHIONE-S-TRANSFERASE AB Methods and reagents is a unique monthly column that highlights current discussions in the newsgroup bionet.molbio.methds-reagnts, available on the Internet. This month's column discusses some common problems associated with the construction of protein fusions with glutathione S-transferase and with hybridization membranes. For details on how to partake in the newsgroup, see the accompanying box. RP Hengen, PN (reprint author), NCI,FREDERICK CANC RES & DEV CTR,FREDERICK,MD 21702, USA. NR 5 TC 8 Z9 8 U1 0 U2 3 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, OXON, ENGLAND OX5 1GB SN 0968-0004 J9 TRENDS BIOCHEM SCI JI Trends Biochem.Sci. PD OCT PY 1996 VL 21 IS 10 BP 400 EP 401 DI 10.1016/S0968-0004(96)30035-2 PG 2 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VN596 UT WOS:A1996VN59600014 PM 8918196 ER PT J AU Szkudlinski, MW Grossmann, M Weintraub, BD AF Szkudlinski, MW Grossmann, M Weintraub, BD TI Structure-function studies of human TSH - New advances in design of glycoprotein hormone analogs SO TRENDS IN ENDOCRINOLOGY AND METABOLISM LA English DT Review ID THYROID-STIMULATING HORMONE; HUMAN CHORIONIC-GONADOTROPIN; RECOMBINANT HUMAN THYROTROPIN; N-LINKED OLIGOSACCHARIDES; BETA-SUBUNIT GENE; ALPHA-SUBUNIT; METABOLIC-CLEARANCE; RECEPTOR-BINDING; SIGNAL-TRANSDUCTION; TERMINAL RESIDUES AB Recent progress in structure-function studies of glycoprotein hormones has provided new insights into the molecular mechanisms of action of these hormones and has further supported the concept that physiological modulation of assembly, bioactivity, and clearance of these hormones is dependent on specific structural components. This review emphasizes current advances in the structure-function relationships of human TSH, which have contributed to further elucidation of common and hormone specific features within the glycoprotein hormones family. Novel strategies are now being applied to investigate the role of individual structural elements. The principles discovered in such studies are essential to understand the physiological regulation of hormone bioactivity and allow for the rational design of novel analogs with potential therapeutic applications. (C) 1996, Elsevier Science Inc. C1 UNIV MARYLAND,SCH MED,DEPT MED,DIV ENDOCRINOL,BALTIMORE,MD 21201. INST HUMAN VIROL,CTR MED BIOTECHNOL,BALTIMORE,MD 21201. RP Szkudlinski, MW (reprint author), NIDDKD,MOL & CELLULAR ENDOCRINOL BRANCH,NIH,BETHESDA,MD 20892, USA. NR 68 TC 22 Z9 23 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 SN 1043-2760 J9 TRENDS ENDOCRIN MET JI Trends Endocrinol. Metab. PD OCT PY 1996 VL 7 IS 8 BP 277 EP 286 DI 10.1016/S1043-2760(96)00129-4 PG 10 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA VM068 UT WOS:A1996VM06800003 PM 18406760 ER PT J AU Wyatt, LS Shors, ST Murphy, BR Moss, B AF Wyatt, LS Shors, ST Murphy, BR Moss, B TI Development of a replication-deficient recombinant vaccinia virus vaccine effective against parainfluenza virus 3 infection in an animal model SO VACCINE LA English DT Article DE parainfluenza virus; vaccinia virus; MVA strain ID RESPIRATORY SYNCYTIAL VIRUS; HEMAGGLUTININ-NEURAMINIDASE; INFLUENZA-VIRUS; PROTECTIVE IMMUNITY; EXPRESSION; TYPE-3; GLYCOPROTEINS; FUSION; VIRULENCE; VECTOR AB The highly attenuated replication-deficient, modified vaccinia virus Ankara (MVA) was used to express the fusion (F) and/or hemagglutinin-neuraminidase (HN) glycoproteins of parainfluenza virus 3 (PIV3). Initial recombinant viruses in which the HN gene was regulated by a very strong synthetic early/late promoter replicated poorly in per missive chick embryo cells evidently due to toxic levels of the gene product. This result led us to construct and evaluate a modified early/late promoter derived from the H5 gene of vaccinia virus. Reporter gene experiments indicated that the enhanced H5 promoter was about Jive times stronger than the 7.5 promoter used in previous recombinant vaccinial PIV3 viruses, Although the overall expression from the modified H5 promoter was less than that of the strong synthetic promoter, early expression, determined in the presence of an inhibitor of DNA replication, was higher. Importantly, recombinant MVA employing the modified H5 promoter to regulate the F or HN gene of PIV3 replicated to high titers in chick cells and expressed functional F or HN proteins as measured by syncytial formation upon dual infection of mammalian cells. Cotton rats inoculated with recombinant MVA expressing F or HN by intramuscular or intranasal routes produced high levels of antibody. The virus expressing HN, however, was the more effective of the two in inducing immunity to PIV3 challenge, reducing PIV3 viral titers in the nasal turbinates by at least 4.7 logs and in the lungs by 3.4 logs, similar to that achieved by immunization with PIV3. These studies support further testing of recombinant MVA/PIV3 viruses as safe and effective candidate vaccines. Copyright (C) 1996 Elsevier Science Ltd. C1 NIAID,VIRAL DIS LAB,NATL INST HLTH,BETHESDA,MD 20892. NIAID,INFECT DIS LAB,NATL INST HLTH,BETHESDA,MD 20892. NR 33 TC 104 Z9 106 U1 0 U2 0 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, OXON, ENGLAND OX5 1GB SN 0264-410X J9 VACCINE JI Vaccine PD OCT PY 1996 VL 14 IS 15 BP 1451 EP 1458 DI 10.1016/S0264-410X(96)00072-2 PG 8 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA WA719 UT WOS:A1996WA71900009 PM 8994321 ER PT J AU Koller, R Krall, M Mock, B Bies, J Nazarov, V Wolff, L AF Koller, R Krall, M Mock, B Bies, J Nazarov, V Wolff, L TI Mml1, a new common integration site in murine leukemia virus-induced promonocytic leukemias maps to mouse chromosome 10 SO VIROLOGY LA English DT Article ID T-CELL LYMPHOMAS; C-MYB ACTIVATION; RETROVIRAL INTEGRATION; PROVIRUS INSERTION; TUMOR PROGRESSION; BALB/C MICE; GENE; LOCUS; LEUKEMOGENESIS; SUSCEPTIBILITY AB MuLV-induced myeloid leukemias (MML) having promonocytic characteristics are produced with high incidence in some strains of adult mice that are undergoing chronic peritoneal inflammation. Previously we showed that many leukemias have rearrangements of the c-myb locus due to insertional mutagenesis, however, we also identified a number of leukemias that had proviral integrations in the absence of c-myb rearrangement. In the present study, a new locus, Mm/1, was found to be a target of insertional mutagenesis in 10 of the promonocytic leukemias that lacked c-myb alterations. Chromosomal mapping studies, performed using progeny from interspecies backcross mice generated by mating (BALB/cAn X M. spretus)F-1 females to BALB/cAN males, determined that Mm/1 is located on the proximal end of mouse chromosome 10. Interestingly, there were no recombinants between c-myb and Mm/1 in 101 backcross progeny and Mm/1 was mapped approximately 20-25 kb upsteam of c-myb. Interestingly, c-myb mRNA and Myb protein are expressed at levels similar to the levels observed in myeloid progenitor cells, but are not overexpressed. It is anticipated that future experiments will determine whether Mm/1 integration prevents down regulation of c-myb expression or activates another gene on chromosome 10. (C) 1996 Academic Press, Inc. C1 NCI,GENET LAB,BETHESDA,MD 20892. NR 50 TC 11 Z9 11 U1 0 U2 0 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0042-6822 J9 VIROLOGY JI Virology PD OCT 1 PY 1996 VL 224 IS 1 BP 224 EP 234 DI 10.1006/viro.1996.0524 PG 11 WC Virology SC Virology GA VP215 UT WOS:A1996VP21500025 PM 8862417 ER PT J AU Schnolzer, M Rackwitz, HR Gustchina, A Laco, GS Wlodawer, A Elder, JH Kent, SBH AF Schnolzer, M Rackwitz, HR Gustchina, A Laco, GS Wlodawer, A Elder, JH Kent, SBH TI Comparative properties of feline immunodeficiency virus (FIV) and human immunodeficiency virus type 1 (HIV-1) proteinases prepared by total chemical synthesis SO VIROLOGY LA English DT Article ID PHASE PEPTIDE-SYNTHESIS; PROTEASE INHIBITOR; CRYSTAL-STRUCTURE; POL POLYPROTEINS; ACTIVE-SITE; HTLV-III; CATS; INFECTION; AIDS; GAG AB The aspartyl proteinase (PR) encoded by the feline immunodeficiency virus (FIV) was prepared by total chemical synthesis. The 116-amino-acid polypeptide chain was assembled in a stepwise fashion using a Boc chemistry solid-phase peptide synthesis approach and subsequently folded into the biologically active dimeric proteinase. The synthetic enzyme showed proteolytic activity against a variety of different peptide substrates corresponding to putative cleavage sites of the Gag and Gag-Pol polyproteins of FIV. A comparative study with the proteinase of human immunodeficiency virus type 1 (HIV-1) showed that the FIV and HIV-1 enzymes have related but distinct substrate specificities. In particular, HIV-1 PR and FIV PR each show a strong preference for their own MA/CA substrates, despite identical amino acid residues at four of seven positions from P3-P4' of the substrate including an identical MA/CA cleavage site (between Tyr similar to Pro residues), FIV PR also showed a requirement for a longer peptide substrate than HIV-1 PR. Defining the similarities and the differences in the properties of these two retroviral enzymes will have a significant impact on structure-based drug design. (C) 1996 Academic Press, Inc. C1 Scripps Res Inst, DEPT MOL BIOL, LA JOLLA, CA 92037 USA. NCI, FREDERICK CANC RES & DEV CTR, ABL BASIC RES PROGRAM, MACROMOL STRUCT LAB, FREDERICK, MD 21702 USA. Scripps Res Inst, DEPT CELL BIOL, LA JOLLA, CA 92037 USA. GERMAN CANC RES CTR, DIV CELL BIOL 0110, D-69120 HEIDELBERG, GERMANY. FU NCI NIH HHS [N01-CO-74101]; NIGMS NIH HHS [P01 GM48870] NR 38 TC 23 Z9 23 U1 0 U2 0 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0042-6822 J9 VIROLOGY JI Virology PD OCT 1 PY 1996 VL 224 IS 1 BP 268 EP 275 DI 10.1006/viro.1996.0528 PG 8 WC Virology SC Virology GA VP215 UT WOS:A1996VP21500029 PM 8862421 ER PT J AU Watkins, BA Crowley, RW Davis, AE Louie, AT Reitz, MS AF Watkins, BA Crowley, RW Davis, AE Louie, AT Reitz, MS TI Expression of CD26 does not correlate with the replication of macrophage-tropic strains of HIV-1 in T-cell lines SO VIROLOGY LA English DT Article ID IMMUNODEFICIENCY-VIRUS TYPE-1; ENVELOPE V3 LOOP; MONONUCLEAR PHAGOCYTES; ACTIVATION ANTIGEN; INFECTION; GP120; IDENTIFICATION; ENTRY; DETERMINANTS; GLYCOPROTEIN AB Strains of human immunodeficiency virus type 1 (HIV-1) differ significantly in both genetic content and biological properties. One of the earliest discovered differences between HIV-1 strains was divergence in the relative ability of different strains to replicate in either T-cell lines or monocytes/macrophages. This observation has led to the suggestion that molecules present on the surface of HIV-susceptible cells other than CD4 may interact with gp120 in facilitating the entry of HIV-1 into host cell populations. Several reports have suggested that CD26, a cell surface protease expressed on many cells of the immune system including some CD4(+) T-cells and macrophage, may be an accessory molecule for HIV-1 entry. Recently, it has also been reported that the expression of high levels of CD26 correlates with the entry and replication of macrophage-tropic strains of HIV-1 in a T-cell line. In this report, we demonstrate that replication of macrophage-tropic strains of HIV-1 in T-cell lines is independent of CD26 expression, From this observation, we conclude that CD26 plays no role in the entry of HIV-1 into these cells. (C) 1996 Academic Press, Inc. RP Watkins, BA (reprint author), NCI,TUMOR CELL BIOL LAB,NIH,BLDG 37,BETHESDA,MD 20892, USA. NR 37 TC 3 Z9 3 U1 0 U2 0 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0042-6822 J9 VIROLOGY JI Virology PD OCT 1 PY 1996 VL 224 IS 1 BP 276 EP 280 DI 10.1006/viro.1996.0529 PG 5 WC Virology SC Virology GA VP215 UT WOS:A1996VP21500030 PM 8862422 ER PT J AU Marlhens, F Bareil, C Griffoin, JM Redmond, TM Arnaud, B Claustres, M Hamel, C AF Marlhens, F Bareil, C Griffoin, JM Redmond, TM Arnaud, B Claustres, M Hamel, C TI Screen for mutations in the RPE65 gene in patients with leber congenital amaurosis (LCA). SO VISION RESEARCH LA English DT Meeting Abstract C1 UNIV MONTPELLIER,INSERM U254,F-34059 MONTPELLIER,FRANCE. UNIV MONTPELLIER,BIOCHIM GENET LAB,F-34059 MONTPELLIER,FRANCE. UNIV MONTPELLIER,DEPT OPHTHALMOL,F-34059 MONTPELLIER,FRANCE. CHU MONTPELLIER,MONTPELLIER,FRANCE. NIH,RETINAL CELL & MOL BIOL LAB,BETHESDA,MD 20892. RI BAREIL, Corinne/A-8387-2008 NR 0 TC 0 Z9 0 U1 0 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, ENGLAND OX5 1GB SN 0042-6989 J9 VISION RES JI Vision Res. PD OCT PY 1996 VL 36 SU S BP 438 EP 438 PG 1 WC Neurosciences; Ophthalmology SC Neurosciences & Neurology; Ophthalmology GA VR898 UT WOS:A1996VR89800616 ER PT J AU Solomon, AS Lacot, JL Rosner, M AF Solomon, AS Lacot, JL Rosner, M TI Corneal findings in rabbits with inherited glaucoma. SO VISION RESEARCH LA English DT Meeting Abstract C1 NEI,NIH,BETHESDA,MD 20892. TEL AVIV UNIV,GOLDSCHLEGER EYE RES INST,IL-69978 TEL AVIV,ISRAEL. NR 0 TC 0 Z9 0 U1 0 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, ENGLAND OX5 1GB SN 0042-6989 J9 VISION RES JI Vision Res. PD OCT PY 1996 VL 36 SU S BP 1415 EP 1415 PG 1 WC Neurosciences; Ophthalmology SC Neurosciences & Neurology; Ophthalmology GA VR898 UT WOS:A1996VR89800085 ER PT J AU GuexCrosier, Y Hayashi, S Delvaux, A Velu, T Roberge, F AF GuexCrosier, Y Hayashi, S Delvaux, A Velu, T Roberge, F TI Downregulation of uveitis by IV IL-10 injection in C3H/HeN mice. SO VISION RESEARCH LA English DT Meeting Abstract C1 NEI,NIH,BETHESDA,MD 20892. UNIV LAUSANNE,DEPT OPHTHALMOL,LAUSANNE,SWITZERLAND. FREE UNIV BRUSSELS,INST RECH INTERDISCIPLINAIRE,BRUSSELS,BELGIUM. FREE UNIV BRUSSELS,DEPT MED GENET,BRUSSELS,BELGIUM. PAOLISTA SCH MED,DEPT OPHTHALMOL,SAO PAULO,BRAZIL. NR 0 TC 0 Z9 0 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, ENGLAND OX5 1GB SN 0042-6989 J9 VISION RES JI Vision Res. PD OCT PY 1996 VL 36 SU S BP 3413 EP 3413 PG 1 WC Neurosciences; Ophthalmology SC Neurosciences & Neurology; Ophthalmology GA VR898 UT WOS:A1996VR89800275 ER PT J AU Schuette, DG Chepelinsky, AB Dilsiz, N Crabbe, MJC AF Schuette, DG Chepelinsky, AB Dilsiz, N Crabbe, MJC TI Insertion and topology of lens MIP in the E-coli cytoplasmic membrane SO VISION RESEARCH LA English DT Meeting Abstract C1 UNIV READING,SCH ANIM & MICROBIAL SCI,READING RG6 2AH,BERKS,ENGLAND. NEI,NIH,BETHESDA,MD 20892. FIRAT UNIV,DEPT BIOL,ELAZIG,TURKEY. NR 0 TC 0 Z9 0 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, ENGLAND OX5 1GB SN 0042-6989 J9 VISION RES JI Vision Res. PD OCT PY 1996 VL 36 SU S BP 3425 EP 3425 PG 1 WC Neurosciences; Ophthalmology SC Neurosciences & Neurology; Ophthalmology GA VR898 UT WOS:A1996VR89800285 ER PT J AU DeMicheli, E Chang, MCJ Rapoport, SI AF DeMicheli, E Chang, MCJ Rapoport, SI TI In vivo imaging of cortical membrane remodeling in rats with chronic unilateral ablation of nucleus basalis magnocellularis: Use of radiolabeled palmitic acid SO BRAIN RESEARCH LA English DT Article DE nucleus basalis magnocellularis; acetylcholine; rat; lesion; ablation; palmitate; neuroplasticity; membrane; remodeling; Alzheimer disease; phospholipid; phosphatidylcholine ID CHOLINE-ACETYLTRANSFERASE; BRAIN INCORPORATION; CEREBRAL-CORTEX; GLUCOSE-UTILIZATION; ALZHEIMERS-DISEASE; FATTY-ACIDS; LESIONS; FOREBRAIN; NEURONS; METABOLITES AB Membrane remodeling was imaged in vivo in brains of rats with a 2-week-old right-sided ablation of the nucleus basalis magnocellularis (NBM). To do this, [9,10-H-3]palmitic acid ([H-3]PAM) was injected intravenously and regional brain incorporation k* of tracer was determined with quantitative autoradiography after 20 min circulation. In NBM-lesioned animals, k* was elevated significantly (by up to 17%) in ii ipsilateral frontal or parietal cortical regions, more so in layer I than in layers TV and V. Unoperated animals showed no right-left difference in k*, whereas sham-operated animals showed some unilateral effects of damage due to the needle track. Circulating [H-3]PAM is incorporated into sn-l positions of brain phospholipids, mainly phosphatidylcholine, and its rate of turnover is thought to reflect turnover of neuronal and glial membranes, These results, when related to published evidence of altered cortical phospholipid metabolism in NBM-lesioned rats, suggest that images of increased [H-3]PAM incorporation into ipsilateral cortex reflect increased membrane remodeling involving phospholipids. C1 NIA,NEUROSCI LAB,NIH,BETHESDA,MD 20892. NR 40 TC 3 Z9 3 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0006-8993 J9 BRAIN RES JI Brain Res. PD SEP 30 PY 1996 VL 735 IS 1 BP 36 EP 41 PG 6 WC Neurosciences SC Neurosciences & Neurology GA VM547 UT WOS:A1996VM54700005 PM 8905167 ER PT J AU Arai, R Jacobowitz, DM Nagatsu, I AF Arai, R Jacobowitz, DM Nagatsu, I TI Calretinin is differentially localized in magnocellular oxytocin neurons of the rat hypothalamus. A double-labeling immunofluorescence study SO BRAIN RESEARCH LA English DT Article DE calretinin; oxytocin; vasopressin; supraoptic nucleus; paraventricular nucleus; anterior commissural nucleus; suprachiasmatic nucleus ID CALCIUM-BINDING PROTEIN; NERVOUS-SYSTEM; IMMUNOHISTOCHEMICAL LOCALIZATION; PARAVENTRICULAR NUCLEUS; SUPRAOPTIC NEURONS; SPINAL-CORD; CALBINDIN-D28K; PARVALBUMIN; CELLS; BRAIN AB By use of a double-labeling immunofluorescence method with a confocal laser scanning microscope, we have examined whether a calcium-binding protein, calretinin, is localized in magnocellular oxytocin and vasopressin neurons of the rat hypothalamus. In the supraoptic nucleus, all oxytocin-labeled cells were stained for calretinin. However, in the magnocellular part of the paraventricular nucleus, almost all oxytocin-stained cells were devoid of calretinin immunoreactivity. All vasopressin-positive cells of both the supraoptic nucleus and the magnocellular part of the paraventricular nucleus lacked calretinin immunoreactivity. No calretinin immunoreactivity was found in oxytocin-labeled cells of the the anterior commissural nucleus or in vasopressin-labeled cells of the suprachiasmatic nucleus. We previously showed that another calcium-binding protein, calbindin-D28k, was localized in magnocellular oxytocin neurons of the supraoptic nucleus but not in those of the paraventricular nucleus. These findings suggest that, in general, magnocellular oxytocin neurons of the supraoptic nucleus and those of the paraventricular nucleus can be chemically distinguished, that is, the former contain both calretinin and calbindin-D28k but the latter lack the two calcium-binding proteins. C1 NIMH,CLIN SCI LAB,BETHESDA,MD 20892. RP Arai, R (reprint author), FUJITA HLTH UNIV,SCH MED,DEPT ANAT,TOYOAKE,AICHI 47011,JAPAN. NR 41 TC 12 Z9 12 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0006-8993 J9 BRAIN RES JI Brain Res. PD SEP 30 PY 1996 VL 735 IS 1 BP 154 EP 158 DI 10.1016/S0006-8993(96)00558-6 PG 5 WC Neurosciences SC Neurosciences & Neurology GA VM547 UT WOS:A1996VM54700019 PM 8905181 ER PT J AU Contois, JH LammiKeefe, CJ Vogel, S McNamara, JR Wilson, PWF Massov, T Schaefer, EJ AF Contois, JH LammiKeefe, CJ Vogel, S McNamara, JR Wilson, PWF Massov, T Schaefer, EJ TI Plasma lipoprotein(a) distribution in the Framingham Offspring Study as determined with a commercially available immunoturbidimetric assay SO CLINICA CHIMICA ACTA LA English DT Article DE lipoprotein(a); lipids; lipoproteins; cholesterol; coronary heart disease; epidemiology ID CORONARY HEART-DISEASE; INDEPENDENT RISK FACTOR; LOW-DENSITY-LIPOPROTEIN; MYOCARDIAL-INFARCTION; ARTERY DISEASE; LP(A) LIPOPROTEIN; APOLIPOPROTEIN-B; TRIGLYCERIDE INTERFERENCE; CEREBROVASCULAR-DISEASE; CEREBRAL INFARCTION AB The purpose of our research was to evaluate a commercially available, automated, immunoturbidimetric assay for lipoprotein(a) (Lp(a)), to determine the distribution of Lp(a) in the Framingham Offspring Study population, and to determine Lp(a) levels that may be useful for assessing coronary heart disease risk. The mean between-run coefficient of variation for this assay was 5.65%. Lp(a) concentration was slightly, but significantly, higher in 1949 white women (mean +/- S,D. 214 +/- 195 mg/l, median 150 mg/l) than in 1884 white men (mean +/- S.D, 200 +/- 193 mg/l, median 130 mg/l) participating in Cycle 4 of the Framingham Offspring Study (P = 0.0015). Lp(a) values of 300 mg/l and 500 mg/l corresponded to approximately the 75th and 90th percentiles, respectively, for both men and women, and subjects with concentrations greater than or equal to 500 mg/l were more likely to have coronary heart disease than subjects with an Lp(a) concentration less than 300 mg/l (P < 0.05 for men). C1 TUFTS UNIV,USDA,JEAN MAYER HUMAN NUTR RES CTR AGING,LIPID METAB LAB,BOSTON,MA 02111. UNIV CONNECTICUT,DEPT NUTR SCI,STORRS,CT 06269. NHLBI,FRAMINGHAM,MA 01701. FU NHLBI NIH HHS [HV-83-03] NR 59 TC 23 Z9 23 U1 1 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0009-8981 J9 CLIN CHIM ACTA JI Clin. Chim. Acta PD SEP 30 PY 1996 VL 253 IS 1-2 BP 21 EP 35 DI 10.1016/0009-8981(96)06341-3 PG 15 WC Medical Laboratory Technology SC Medical Laboratory Technology GA VG192 UT WOS:A1996VG19200003 PM 8879836 ER PT J AU Ribas, JC Wickner, RB AF Ribas, JC Wickner, RB TI RNA-dependent RNA polymerase activity related to the 20S RNA replicon of Saccharomyces cerevisiae SO YEAST LA English DT Article DE replication; WdsRNA; RNA polymerase ID DOUBLE-STRANDED-RNA; POL FUSION PROTEIN; L-A-VIRUS; GAG-POL; VIRAL-RNA; 20-S RNA; T-DSRNA; YEAST; FORM; IDENTIFICATION AB Saccharomyces cerevisiae contains two double-stranded RNA (dsRNA) viruses (L-A and L-BC) and two different single-stranded (ssRNA) replicons (20S RNA and 23S RNA). Replicase (dsRNA synthesis on a ssRNA template) and transcriptase (ssRNA synthesis on a dsRNA template) activities have been described for L-A and L-BC viruses, but not for 20S or 23S RNA. We report the characterization of a new in vitro RNA replicase activity in S. cerevisiae. This activity is detected after partial purification of a particulate fraction in CsCl gradients where it migrates al the density of free protein. The activity does not require the presence of L-A or L-BC viruses or 23S RNA, and its presence or absence is correlated with the presence or absence of the 20S RNA replicon. Strains lacking both this RNA polymerase activity and 20S RNA acquire this activity when they acquire 20S RNA by cytoduction (cytoplasmic mixing). This polymerase activity converts added ssRNA to dsRNA by synthesis of the complementary strand, but has no specificity for the 3' end or internal template sequence. Although it replicates all tested RNA templates, it has a template size requirement, being unable to replicate templates larger than 1 kb. The replicase makes dsRNA from a ssRNA template, but many single-stranded products due to a terminal transferase activity are also formed. These results suggest that, in contrast to the L-A and L-BC RNA polymerases, dissociation of 20S RNA polymerase from its RNA (or perhaps some cellular factor) makes the enzyme change its specificity. C1 NIDDKD, SECT GENET SIMPLE EUKARYOTES, BETHESDA, MD 20892 USA. RI Ribas, Juan/C-9864-2015 OI Ribas, Juan/0000-0001-6430-0895 NR 37 TC 1 Z9 1 U1 0 U2 0 PU WILEY-BLACKWELL PI HOBOKEN PA 111 RIVER ST, HOBOKEN 07030-5774, NJ USA SN 0749-503X EI 1097-0061 J9 YEAST JI Yeast PD SEP 30 PY 1996 VL 12 IS 12 BP 1219 EP 1228 PG 10 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Microbiology; Mycology SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Microbiology; Mycology GA VN097 UT WOS:A1996VN09700004 PM 8905926 ER PT J AU Badio, B Shi, D Shin, Y Hutchinson, KD Padgett, WL Daly, JW AF Badio, B Shi, D Shin, Y Hutchinson, KD Padgett, WL Daly, JW TI Spiropyrrolizidines: A new class of blockers of nicotinic receptors SO BIOCHEMICAL PHARMACOLOGY LA English DT Article DE cholinergic receptors; noncompetitive antagonists; nicotine ID PHEOCHROMOCYTOMA PC12 CELLS; ACETYLCHOLINE-RECEPTOR; RAT-BRAIN; POISON-FROG; MOUSE-BRAIN; ALKALOIDS; BINDING; CHANNEL; SITES; DENDROBATIDAE AB The spiropyrrolizidine oximes 236 and 222 and a related spiropyrrolizidine alkaloid, nitropolyzonamine, block nicotinic receptor channels in rat pheochromocytoma PC12 cells and in human medulloblastoma TE671 cells. In PC12 cells with an alpha(3) beta(4(5))-nicotinic receptor, both the spiropyrrolizidine oxime 236 and nitropolyzonamine had IC50 values of about 1.5 mu M, while spiropyrrolizidine oxime 222 had an IC50 value of 2.6 mu M versus carbamylcholine-elicited sodium-22 influx. In TE671 cells with an alpha(1) beta(1) gamma delta nicotinic receptor, the spiropyrrolizidine oximes 236, 222, and nitropolyzonamine had IC50 values of 9.5, 14, and 67 mu M, respectively. The inhibitions by the spiropyrrolizidine oxime 236 and nitropolyzonamine appeared to be noncompetitive in nature in both cell lines. In rat cerebral cortical membranes, binding of [H-3]nicotine to alpha(4) beta(2) nicotinic receptors was not inhibited significantly by 10 mu M concentrations of the spiropyrrolizidine oxime 236, or by nitropolyzonamine, as expected for a noncompetitive blocker. Both compounds at 10 mu M had marginal effects on a variety of central receptors, but did inhibit binding of [H-3] 1,3-di(2-tolyl)guanidine to sigma receptors in mouse brain membranes with IC50 values of about 0.5 mu M. The spiropyrrolizidine oxime 236 at 10 mu M had no effect on batrachotoxin-elicited sodium influx in guinea pig cerebral cortical synaptoneurosomes or on ATP-elicited calcium influx in PC12 cells. Such spiropyrrolizidines represent a new structural class of blockers of nicotinic receptor channels with selectivity for ganglionic-type receptors. RP Badio, B (reprint author), NIDDKD,NIH,BIOORGAN CHEM LAB,BLDG 8,RM 1A15,BETHESDA,MD 20892, USA. NR 27 TC 12 Z9 14 U1 0 U2 6 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, ENGLAND OX5 1GB SN 0006-2952 J9 BIOCHEM PHARMACOL JI Biochem. Pharmacol. PD SEP 27 PY 1996 VL 52 IS 6 BP 933 EP 939 DI 10.1016/0006-2952(96)00464-9 PG 7 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA VD957 UT WOS:A1996VD95700016 PM 8781513 ER PT J AU Chua, KL Wiklund, F Lenner, P Angstrom, T Hallmans, G Bergman, F Sapp, M Schiller, J Wadell, G Hjerpe, A Dillner, J AF Chua, KL Wiklund, F Lenner, P Angstrom, T Hallmans, G Bergman, F Sapp, M Schiller, J Wadell, G Hjerpe, A Dillner, J TI A prospective study on the risk of cervical intra-epithelial neoplasia among healthy subjects with serum antibodies to HPV compared with HPV DNA in cervical smears SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article ID HUMAN PAPILLOMAVIRUS INFECTION; POLYMERASE CHAIN-REACTION; VIRUS-LIKE PARTICLES; YOUNG-WOMEN; TYPE-16; CANCER; DYSPLASIA; PRIMERS AB To estimate the risk of developing cervical intra-epithelial neoplasia (CIN) among women exposed to human papillomavirus (HPV) type 16, we performed a prospective study in a population-based cohort of more than 15,000 women followed for 34.9 months. Seventy-four women developed CIN during follow-up and were matched for age, time of sampling and area of residence with 148 women who remained CIN-free during follow-up. The blood samples taken at enrollment were tested for serum antibodies to HPV types 16, 18 and 33 capsids. Cervical smears or biopsies were analyzed for the presence of HPV DNA by nested PCR using HPV general primers and by HPV 16- and 18-type-specific PCR. HPV serology and HPV-PCR were in good agreement, particularly when the blood sample and the Pap smear were taken less than 6 months apart. HPV DNA was found in 88% of cases and 4% of controls, whereas HPV 16 DNA was present in 44% of cases and in 1 of 142 controls. HPV-16-seropositive women had a 3-fold increased risk of developing CIN. The risk was highest among women younger than 35 years of age, of whom an estimated 3.4% of HPV-16-seropositive and 0.5% of seronegative women developed CIN, Since the risk associated with HPV-16 seropositivity (a measure of past or present infection) was 35-fold lower than that of HPV DNA (present infection), most infections appear to be eliminated before CIN develops. In conclusion, HPV 16 infection does confer an excess risk of CIN development, and HPV DNA detection has a high predictive value for the presence of high-grade CIN. (C) 1996 Wiley-Liss, Inc. C1 KAROLINSKA INST,CTR MICROBIOL & TUMOR BIOL,S-17177 STOCKHOLM,SWEDEN. HUDDINGE UNIV HOSP,KAROLINSKA INST,DEPT PATHOL,S-14186 HUDDINGE,SWEDEN. UMEA UNIV,DEPT ONCOL,UMEA,SWEDEN. UMEA UNIV,DEPT CLIN CYTOL,UMEA,SWEDEN. UMEA UNIV,DEPT NUTR RES,UMEA,SWEDEN. UMEA UNIV,DEPT PATHOL,S-90187 UMEA,SWEDEN. UMEA UNIV,DEPT VIROL,UMEA,SWEDEN. UNIV MAINZ,DEPT MED MICROBIOL,D-6500 MAINZ,GERMANY. NATL CANC INST,CELLULAR ONCOL LAB,BETHESDA,MD. NR 30 TC 60 Z9 61 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC 605 THIRD AVE, NEW YORK, NY 10158-0012 SN 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD SEP 27 PY 1996 VL 68 IS 1 BP 54 EP 59 DI 10.1002/(SICI)1097-0215(19960927)68:1<54::AID-IJC11>3.0.CO;2-6 PG 6 WC Oncology SC Oncology GA VK978 UT WOS:A1996VK97800011 PM 8895541 ER PT J AU Ding, M Vitale, N Tsai, SC Adamik, R Moss, J Vaughan, M AF Ding, M Vitale, N Tsai, SC Adamik, R Moss, J Vaughan, M TI Characterization of a GTPase-activating protein that stimulates GTP hydrolysis by both ADP-ribosylation factor (ARF) and ARF-like proteins - Comparison to the ARD1 GAP-domain SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID NUCLEOTIDE-BINDING PROTEINS; GUANINE-NUCLEOTIDE; CHOLERA-TOXIN; PHOSPHOLIPASE-D; BREFELDIN-A; SELECTIVE AMPLIFICATION; ACID PHOSPHOLIPIDS; BOVINE BRAIN; GOLGI; EXCHANGE AB ADP-ribosylation factors (ARFs) are similar to 20-kDa guanine nucleotide-binding proteins that participate in vesicular transport in the Golgi and other intracellular compartments and stimulate cholera toxin ADP-ribosyltransferase activity. Both GTP binding and hydrolysis are necessary for its physiological functions, although purified mammalian ARF lacks detectable GTPa selectivity. An ARF GTPase-activating protein (GAP) was purified >15,000-fold from rat spleen cytosol using (NH4)(2)SO4 precipitation and chromatography on Ultrogel AcA 34, DEAE-Sephacel, heparin-Sepharose, hydroxylapatite, and Ultrogel AcA 44. In fractions (similar to 100-kDa proteins) from Ultrogel AcA 44, a major protein band of similar to 50 kDa on SDS-polyacrylamide gel electrophoresis correlated with GAP activity, consistent with it being a homodimer, thus differing from an ARF GAP purified from rat liver (Makler, V., Cukierman, E., Rotman, M., Admon, A., and Cassel, D. (1995) J. Biol. Chem. 270, 5232-5237). Purified spleen GAP accelerated hydrolysis of GTP bound to recombinant ARF1, ARF3, ARF5, and ARF6; no effect of NH2-terminal myristoylation was observed, ARF GAP also activated GTP hydrolysis by ARL1, which is 56% identical in amino acid sequence to ARF1, but lacks ARF activity, ARD1 is a 64-MDa guanine nucleotide binding protein that contains an 18-kDa ARF domain at its carboxyl terminus; the ARF domain lacks the amino-terminal alpha-helix found in native ARF and hence is similar to the amino-terminal truncated mutant Delta 13ARF1. Both the ARF domain of ARD1 and Delta 13ARF1 were poor substrates for ARF GAP. The non-ARF1 do main of ARD1 enhanced the GTPase activity of the ARF domain, but not that of the ARF proteins and Delta 13ARF1, i.e. it lacks the relatively broad substrate specificity exhibited by ARF GAP. C1 NHLBI, NATL INST HLTH, PULM CRIT CARE MED BRANCH, BETHESDA, MD 20892 USA. RI Vitale, nicolas/G-5967-2014 OI Vitale, nicolas/0000-0002-4752-4907 NR 32 TC 28 Z9 28 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 SEP 27 PY 1996 VL 271 IS 39 BP 24005 EP 24009 PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VJ442 UT WOS:A1996VJ44200064 PM 8798635 ER PT J AU Jang, SI Steinert, PM Markova, NG AF Jang, SI Steinert, PM Markova, NG TI Activator protein 1 activity is involved in the regulation of the cell type-specific expression from the proximal promoter of the human profilaggrin gene SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HUMAN PAPILLOMAVIRUS TYPE-18; DIFFERENTIATION-SPECIFIC EXPRESSION; KERATINOCYTE-SPECIFIC TRANSCRIPTION; EPIDERMAL DIFFERENTIATION; TRANSGENIC MICE; DNA-BINDING; HUMAN LORICRIN; RETINOIC ACID; AP-1 ELEMENT; AP1 SITES AB The human profilaggrin gene is expressed in the granular layer during the late stages of terminal differentiation of the epidermis. In in vitro transcription experiments we show that the abundance of the mRNA and the specificity of the expression are regulated primarily at the level of transcription. We found that the 5'-flanking sequences control the transcription in a keratinocyte-specific mode and that as little as 116 base pairs preceding the mRNA initiation site is sufficient to restrict the transcription to epidermal cells in vitro. This specificity depends critically on the presence of an activator protein 1 (AP1) motif at position -77. Binding of c-jun/c-fos heterodimers to this sequence confers high levels of expression to the reporter constructs in cultured epidermal keratinocytes, while having little effect in HeLa cells. The transactivating properties of c-jun are essential in this process. On the other hand, junB and junD, which are involved in transactivating the transcription of earlier epidermal differentiation markers, control profilaggrin expression through a pathway which does not depend on a direct binding at the AP1 site and is not cell-type specific. These data indicate that AP1 factors are involved in a complex, multipathway regulation of the profilaggrin gene expression. C1 NIAMS,NATL INST HLTH,SKIN BIOL LAB,BETHESDA,MD 20892. NR 71 TC 78 Z9 78 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 SEP 27 PY 1996 VL 271 IS 39 BP 24105 EP 24114 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VJ442 UT WOS:A1996VJ44200078 PM 8798649 ER PT J AU Kim, HY LaVaute, T Iwai, K Klausner, RD Rouault, TA AF Kim, HY LaVaute, T Iwai, K Klausner, RD Rouault, TA TI Identification of a conserved and functional iron-responsive element in the 5'-untranslated region of mammalian mitochondrial aconitase SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID BINDING-PROTEIN; RNA-BINDING; REGULATORY FACTOR; SULFUR CLUSTER; MESSENGER-RNA; TRANSLATIONAL REPRESSOR; SUCCINATE-DEHYDROGENASE; CYTOSOLIC ACONITASE; SUBUNIT; GENE AB Iron responsive elements (IREs) are RNA stem-loop motifs found in genes of iron metabolism. When cells are iron depleted, iron regulatory proteins (IRPs) bind to IREs in the transcripts of ferritin, transferrin receptor, and erythroid amino-levulinic acid synthetase. Binding of IRPs to IRE motifs near the 5' end of the transcript results in attenuation of translation while binding to IREs in the 3'-untranslated region of the transferrin receptor results in protection from endonucleolytic cleavage. Iron deprivation results in activation of IRE binding activity, whereas iron replete cells lose IRE binding activation. Here, we report the identification of a conserved IRE in the 5'-untranslated region of the transcript of the citric acid cycle enzyme mitochondrial aconitase from four different mammalian species. The IRE in the transcript of mitochondrial aconitase can mediate in vitro translational repression of mitochondrial aconitase by IRPs. Furthermore, levels of mitochondrial aconitase are decreased in mice maintained on a low iron diet, whereas levels of mRNA remain unchanged. The decrease in levels of mitochondrial aconitase is likely due to activation of IRP binding and consequent attenuation of translation. Thus, expression of the iron-sulfur protein mitochondrial aconitase and function of the citric acid cycle may be regulated by iron levels in cells. RP Kim, HY (reprint author), NICHHD,CELL BIOL & METAB BRANCH,NATL INST HLTH,BETHESDA,MD 20892, USA. NR 27 TC 90 Z9 91 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 J9 J BIOL CHEM JI J. Biol. Chem. PD SEP 27 PY 1996 VL 271 IS 39 BP 24226 EP 24230 PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VJ442 UT WOS:A1996VJ44200095 PM 8798666 ER PT J AU Blough, BE Abraham, P Lewin, AH Kuhar, MJ Boja, JW Carroll, FI AF Blough, BE Abraham, P Lewin, AH Kuhar, MJ Boja, JW Carroll, FI TI Synthesis and transporter binding properties of 3 beta-(4'-alkyl-, 4'-alkenyl-, and 4'-alkynylphenyl)nortropane-2 beta-carboxylic acid methyl esters: Serotonin transporter selective analogs SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID DOPAMINE UPTAKE SITES; COCAINE BINDING; NOREPINEPHRINE TRANSPORTERS; RAT STRIATUM; INHIBITION; AFFINITY; RECEPTORS; REAGENTS; PUTAMEN AB New methods for the synthesis of 3 beta-(4'-alkyl-, 4'-alkenyl-, and 4'-alkynylphenyl)nortropane-2 beta-carboxylic acid methyl esters 2-4, respectively, were developed. These methods involved coupling of the appropriate organometallic reagents to 3 beta-(4'-iodophenyl)tropane-2 beta-carboxylic acid methyl ester (6a, RTI-55) or to an N-protected derivative of 6a followed by N-demethylation or removal of the protecting group. Some analogs were prepared by catalytic reduction of the alkene and alkyne analogs 3 and 4 or by isomerization of the alkenes 3. The analogs 2-4 were evaluated for inhibition of radioligand binding to the serotonin (5-HT), dopamine (DA), and norepinephrine (NE) transporters. 3 beta-(4'-Isopropenyl- and 4'-cis-propenylphenyl)nortropane-2 beta-carboxylic acid methyl esters (3b,d), which possessed IC50 values of 0.6 and 1.15 nM, respectively, were the most potent analogs at the 5-HT transporter, and with NE/5-HT IC50 ratios of 240 and 128 nM, respectively, they were selective for the 5-HT relative to the NE transporter. Since interaction with the serotonin transporter may modulate the pharmacological effects resulting from binding to the dopamine transporter, 3 beta-(4'-isopropenylphenyl)tropane-2 beta-carboxylic acid methyl ester (11b) which has good affinity for both the 5-HT and DA transporters but low affinity at the NE transporter may be useful for studying this interaction. C1 RES TRIANGLE INST,RES TRIANGLE PK,NC 27709. NIDA,ADDICT RES CTR,NEUROSCI BRANCH,BALTIMORE,MD 21224. FU NIDA NIH HHS [DA05477] NR 28 TC 54 Z9 54 U1 2 U2 2 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 SN 0022-2623 J9 J MED CHEM JI J. Med. Chem. PD SEP 27 PY 1996 VL 39 IS 20 BP 4027 EP 4035 DI 10.1021/jm960409s PG 9 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA VK234 UT WOS:A1996VK23400018 PM 8831768 ER PT J AU Khamnei, S Torrence, PF AF Khamnei, S Torrence, PF TI Neighboring group catalysis in the design of nucleotide prodrugs SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; DERIVATIVES; ZIDOVUDINE; DELIVERY; SYSTEM; BRAIN; 3'-AZIDO-3'-DEOXYTHYMIDINE; PHOSPHATE; ANALOGS; AIDS AB An approach is described for potential application to the delivery of polar nucleosides and nucleotides across lipophilic membranes, namely, nucleotide prodrugs based on salicyl phosphate. 3'-Azido-3'-deoxythymidine (AZT) and 3'-deoxythymidine (ddT) were chosen as models. For the synthesis of prototype compounds 1 and 2, the approach was first to react either methyl salicylate (for 1) or phenyl salicylate (for 2) with phosphorus oxychloride in dry methylene chloride at 0 degrees C with the addition of triethylamine as acid scavenger. The resulting intermediate phosphorodichloridate was reacted immediately with excess nucleoside under the same conditions. The control model compound 3 was prepared by reaction of phenyl phosphorodichloridate and excess nucleoside in pyridine/methylene chloride at 0 degrees C to give 3 in 82% yield. The synthesis of triester 7 involved reaction of alpha-(chloroacetyl)salicyl chloride with 2,3,4,6-tetra-O-benzyl-D-glucopyranose to give [[(2,3,4,6-tetra-O-benzyl-D-glucopyranosyl)oxy]carbonyl]-2-(1-chloroacetoxy)benzene (4) which was dechloroacetylated to 5, 2,3,4,6-tetra-O-benzyl-D-glucopyranosyl salicylate. Phosphorylation of 5 with phosphorus oxychloride provided the phosphorodichloridate which was directly converted to 6 by reaction with dideoxythymidine. Removal of benzyl groups by catalytic hydrogenation gave compound 7, bis(2',3'-dideoxythymidin-5'-yl) D-glucopyranosyl phosphate. The AZT prodrug triesters, 1 and 2, underwent much more rapid hydrolysis than the triester 3, most probably due to the formation of an acyl phosphate complex from the attack on phosphorus of the salicylate carboxylate. The hydrolysis of the less lipophilic 7 was significantly slower than that of 1 or 2. Both pig liver esterase and rat brain cytosol were able to effect the cleavage to dinucleotide or mononucleotide of prodrug forms 2 and 7, much more rapidly than either 3 or 1, suggesting that the esterase-like enzymatic activity of rat brain was similar to that of pig liver esterase. This study suggests the possibility of use of salicylic acid-based prodrugs for nucleotides, subject to specific refinements in the choice of carboxylate- and phosphoric acid ester-protecting groups. C1 NIDDKD,MED CHEM LAB,BIOMED CHEM SECT,MED CHEM LAB,NIH,BETHESDA,MD 20892. NR 23 TC 8 Z9 8 U1 0 U2 10 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 SN 0022-2623 J9 J MED CHEM JI J. Med. Chem. PD SEP 27 PY 1996 VL 39 IS 20 BP 4109 EP 4115 DI 10.1021/jm9600757 PG 7 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA VK234 UT WOS:A1996VK23400026 PM 8831776 ER PT J AU Dean, M Carrington, M Winkler, C Huttley, GA Smith, MW Allikmets, R Goedert, JJ Buchbinder, SP Vittinghoff, E Gomperts, E Donfield, S Vlahov, D Kaslow, R Saah, A Rinaldo, C Detels, R OBrien, SJ AF Dean, M Carrington, M Winkler, C Huttley, GA Smith, MW Allikmets, R Goedert, JJ Buchbinder, SP Vittinghoff, E Gomperts, E Donfield, S Vlahov, D Kaslow, R Saah, A Rinaldo, C Detels, R OBrien, SJ TI Genetic restriction of HIV-1 infection and progression to AIDS by a deletion allele of the CKR5 structural gene SO SCIENCE LA English DT Article ID IMMUNODEFICIENCY-VIRUS TYPE-1; MOLECULAR-CLONING; RECEPTOR; DNA; POLYMORPHISMS; LOCALIZATION; POPULATIONS; HEMOPHILIA; EVOLUTION; COHORT AB The chemokine receptor 5 (CKR5) protein serves as a secondary receptor on CD4(+) T lymphocytes for certain strains of human immunodeficiency virus-type 1 (HIV-1). The CKR5 structural gene was mapped to human chromosome 3p21, and a 32-base pair deletion allele (CKR5 Delta 32) was identified that is present at a frequency of similar to 0.10 in the Caucasian population of the United States. An examination of 1955 patients included among six well-characterized acquired immunodeficiency syndrome (AIDS) cohort studies revealed that 17 deletion homozygotes occurred exclusively among 612 exposed HIV-1 antibody-negative individuals (2.8 percent) and not at all in 1343 HIV-1-infected individuals. The frequency of CKR5 deletion heterozygotes was significantly elevated in groups of individuals that had survived HIV-1 infection for more than 10 years, and, in some risk groups, twice as frequent as their occurrence in rapid progressors to AIDS. Survival analysis clearly shows that disease progression is slower in CKR5 deletion heterozygotes than in individuals homozygous for the normal CKR5 gene. The CKR5 Delta 32 deletion may act as a recessive restriction gene against HIV-1 infection and may exert a dominant phenotype of delaying progression to AIDS among infected individuals. C1 NCI,LAB GENOM DIVERS,FREDERICK,MD 21702. NCI,INTRAMURAL RES SUPPORT PROGRAM,SCI APPLICAT INT CORP FREDERICK,FREDERICK,MD 21702. NCI,VIRAL EPIDEMIOL BRANCH,BETHESDA,MD 20892. SAN FRANCISCO CITY CLIN,DEPT PUBL HLTH,SAN FRANCISCO,CA 94102. CHILDRENS HOSP LOS ANGELES,LOS ANGELES,CA 90027. NEW ENGLAND RES INST INC,WATERTOWN,MA 02172. JOHNS HOPKINS UNIV,SCH HYG & PUBL HLTH,DEPT EPIDEMIOL,BALTIMORE,MD 21205. UNIV ALABAMA,DEPT EPIDEMIOL,BIRMINGHAM,AL 35294. UNIV PITTSBURGH,SCH PUBL HLTH,PITTSBURGH,PA 15213. UNIV CALIF LOS ANGELES,SCH PUBL HLTH,LOS ANGELES,CA 90025. UNIV CALIF LOS ANGELES,SCH MED,LOS ANGELES,CA 90025. RI Smith, Michael/B-5341-2012; Dean, Michael/G-8172-2012; Huttley, Gavin/G-5169-2015 OI Dean, Michael/0000-0003-2234-0631; Huttley, Gavin/0000-0001-7224-2074 FU NCRR NIH HHS [MO1-RR06020]; NICHD NIH HHS [N01-HD-4-3200]; NIDA NIH HHS [DA04334] NR 55 TC 1881 Z9 1944 U1 13 U2 79 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 SN 0036-8075 J9 SCIENCE JI Science PD SEP 27 PY 1996 VL 273 IS 5283 BP 1856 EP 1862 DI 10.1126/science.273.5283.1856 PG 7 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VJ713 UT WOS:A1996VJ71300044 PM 8791590 ER PT J AU McCann, UD Hatzidimitriou, G Ricaurte, GA AF McCann, UD Hatzidimitriou, G Ricaurte, GA TI Prolactin response to fenfluramine is independent of serotonin release SO EUROPEAN JOURNAL OF PHARMACOLOGY LA English DT Article DE 5-HT (5-hydroxytryptamine, serotonin); fluoxetine; neuroendocrine challenge ID 5-HYDROXYTRYPTAMINE; NEUROTOXICITY; HYPOPHAGIA AB To assess the role of serotonin release in the prolactin response to fenfluramine, rats were treated with fenfluramine alone or in combination with a dose of fluoxetine known to block fenfluramine-induced serotonin release. Fluoxetine pretreatment did not prevent fenfluramine-induced increases in prolactin. These findings indicate that fenfluramine-induced increases in prolactin are independent of serotonin release, and possibly involve direct post-synaptic actions of fenfluramine or one of its metabolites (norfenfluramine). C1 JOHNS HOPKINS MED INST,DEPT NEUROL,BALTIMORE,MD 21224. NIMH,UNIT ANXIETY DISORDERS,BIOL PSYCHIAT BRANCH,NIH,BETHESDA,MD 20892. FU NIDA NIH HHS [K02 DA00206, R01 DA06275] NR 8 TC 12 Z9 11 U1 0 U2 0 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 SEP 26 PY 1996 VL 312 IS 2 BP R1 EP R2 DI 10.1016/0014-2999(96)00596-1 PG 2 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA VK388 UT WOS:A1996VK38800021 PM 8894606 ER PT J AU Ratty, AK Jeong, SW Nagle, JW Chin, H Gainer, H Murphy, D Venkatesh, B AF Ratty, AK Jeong, SW Nagle, JW Chin, H Gainer, H Murphy, D Venkatesh, B TI A systematic survey of the intergenic region between the murine oxytocin- and vasopressin-encoding genes SO GENE LA English DT Article DE transcription factor recognition sequences; repetitive elements; 129/SvJ and C57BL/10A mouse strains ID PROTEIN CODING REGIONS; TRANSGENIC MICE; TRANSCRIPTION FACTOR; MAGNOCELLULAR NEURONS; RAT VASOPRESSIN; EXPRESSION; PROMOTER; SEQUENCE; ENHANCER; BINDING AB The genomic region between the oxytocin (OT) and vasopressin (VP) genes in the two strains of mice was independently sequenced by our two groups. In this report, we present our collated sequence data and analyses. The mouse intergenic region (MUIGR) was aligned to that of the rat, which has been reported to contain 6.4-kb long interspersed nuclear element (LINE). The MUIGR sequences in the two mice strains did not contain any LINE sequences. This suggests that the approximately 3.5-kb sequence that is conserved between the rat and mouse intergenic regions is likely to be involved in the regulation of OT and VP expression. We also observed several conserved putative transcription factor recognition sequences. Analysis of the MUIGR revealed the lack of any significant ORFs, but the presence of several repetitive elements. C1 NATL UNIV SINGAPORE, INST MOL & CELL BIOL, SINGAPORE 117548, SINGAPORE. NINCDS, NEUROSCI LAB, NIH, BETHESDA, MD 20892 USA. NINCDS, NEUROGENET SECT, NIH, BETHESDA, MD 20892 USA. RI Murphy, David/C-3967-2012; ASTAR, IMCB/E-2320-2012; OI Murphy, David/0000-0003-2946-0353; Venkatesh, Byrappa/0000-0003-3620-0277 NR 44 TC 19 Z9 19 U1 0 U2 0 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 SEP 26 PY 1996 VL 174 IS 1 BP 71 EP 78 DI 10.1016/0378-1119(96)00370-8 PG 8 WC Genetics & Heredity SC Genetics & Heredity GA VL049 UT WOS:A1996VL04900011 PM 8863731 ER PT J AU Chen, D Guo, JR Miki, T Tachibana, M Gahl, WA AF Chen, D Guo, JR Miki, T Tachibana, M Gahl, WA TI Molecular cloning of two novel rab genes from human melanocytes SO GENE LA English DT Article DE GTPase; melanosome; dense body; lysosome; vesicles; membranes ID GTP-BINDING PROTEINS; EPITHELIAL-CELLS; GTPASES; FAMILY AB We isolated the genes of two small GTP-binding proteins of the rab family from a human melanocyte cDNA library and from melanoma cells. One gene, rab30 codes for a novel rab protein of 203 amino acids with minimal homology to previously documented GTPases. The other, rab22b, appears to be an isoform of the human homologue of canine rab22. Both rab mRNAs displayed a nearly ubiquitous pattern of expression in the various tissues examined. Rab22b and rab30 were mapped to chromosomes 18 and 11, respectively. C1 NICHHD,HUMAN BIOCHEM GENET SECT,HERITABLE DISORDERS BRANCH,NIH,BETHESDA,MD 20892. NCI,CELLULAR & MOL BIOL LAB,NIH,BETHESDA,MD 20892. NIH,NATL INST DEAFNESS & OTHER COMMUN DISORDERS,MOL GENET LAB,BETHESDA,MD 20892. NR 19 TC 31 Z9 34 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 SEP 26 PY 1996 VL 174 IS 1 BP 129 EP 134 DI 10.1016/0378-1119(96)00509-4 PG 6 WC Genetics & Heredity SC Genetics & Heredity GA VL049 UT WOS:A1996VL04900019 PM 8863739 ER PT J AU Funkenstein, B Jakowlew, SB AF Funkenstein, B Jakowlew, SB TI Molecular cloning of fish alcohol dehydrogenase cDNA SO GENE LA English DT Article DE recombinant DNA; nucleotide and amino acid sequence; RNA blot hybridization; RT-PCR; fish; larvae ID DEPENDENT FORMALDEHYDE DEHYDROGENASE; CLASS-III; CLASS-I; GENE; EXPRESSION; FAMILY; SEQUENCE; ORIGIN; ADH AB A cDNA encoding a putative alcohol dehydrogenase class III (ADH) was cloned from a cDNA library constructed from 7-day larvae RNA of the marine teleost Sparus aurata. The full length cDNA is 1350 nucleotides (nt) long and contains an ORF of 1128 nt [encoding 376 amino acid (aa) residues]. Identity of 82% was found with human class III ADH (305 of 372 aa compared), and only 62% identity with a fish (cod) ADH (234 of 375 aa compared). Northern hybridization analysis with the cDNA revealed a transcript of about 1.4-1.5 kb, which is expressed in all tissues from adult fish studied: skeletal muscle, heart muscle, kidney, gill filaments and liver, with the highest levels found in the kidney. The expression of ADH mRNA was determined also during early development of Sparus aurata by Northern blot analysis. ADH transcripts were detected in eggs and in embryos 4, 8 and 12 h after fertilization, as well as on all days post-hatching studied. The levels of expression decreased during early embryonal development, but increased 4-fold from day 1 to day 21 after hatching. The size of the transcript was identical to that of hepatic ADH. Our results suggest that maternal ADH mRNA is present in the eggs and embryos, which decreases as divisions and development occur, while after hatching ADH mRNA is expressed by the larval tissues. C1 NCI,BIOMARKERS & PREVENT RES BRANCH,ROCKVILLE,MD 20850. RP Funkenstein, B (reprint author), NATL INST OCEANOG,ISRAEL OCEANOG & LIMNOL RES,TEL SHIKMONA,POB 8030,IL-31080 HAIFA,ISRAEL. NR 21 TC 11 Z9 13 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-1119 J9 GENE JI Gene PD SEP 26 PY 1996 VL 174 IS 1 BP 159 EP 164 DI 10.1016/0378-1119(96)00513-6 PG 6 WC Genetics & Heredity SC Genetics & Heredity GA VL049 UT WOS:A1996VL04900023 PM 8863743 ER PT J AU Hsich, G Kinney, K Gibbs, CJ Lee, KH Harrington, MG AF Hsich, G Kinney, K Gibbs, CJ Lee, KH Harrington, MG TI The 14-3-3 brain protein in cerebrospinal fluid as a marker for transmissible spongiform encephalopathies SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Article ID CREUTZFELDT-JAKOB-DISEASE; GROWTH-HORMONE THERAPY; 2-DIMENSIONAL ELECTROPHORESIS; 14-3-3-PROTEINS; DIAGNOSIS; KINASE; ACTIVATION; TISSUE; GELS; RAF AB Background There is no practical and reliable premortem test for Creutzfeldt-Jakob disease and the related transmissible spongiform encephalopathies. Two proteins, designated 130 and 131, which have been detected in low concentrations in cerebrospinal fluid from patients with Creutzfeldt-Jakob disease, appear to be sensitive and specific markers for the disease. Attempts to identify these proteins, however, have been unsuccessful. We hypothesized that they may be present in the normal brain. Methods We detected proteins 130 and 131 in normal human brain, partially sequenced their amino acids, and found that they matched the brain protein known as 14-3-3. We then developed a simple, rapid immunoassay for this protein and tested it in cerebrospinal fluid samples from 71 humans and 30 animals with spongiform encephalopathies and in control samples from 186 humans and 94 animals. Results The immunoassay detected the 14-3-3 protein in cerebrospinal fluid from 68 of the 71 patients with Creutzfeldt-Jakob disease (96 percent; 95 percent confidence interval, 92 to 99 percent). Among 94 patients with other dementias, the specificity was 96 percent. If one excludes the three patients with dementia who had had strokes within one month before testing, the specificity was 99 percent. The test was positive in 12 of 24 patients with viral encephalitis. In animals the sensitivity of the assay was 87 percent and the specificity was 99 percent. Conclusions In patients with dementia, a positive immunoassay for the 14-3-3 brain protein in cerebrospinal fluid strongly supports a diagnosis of Creutzfeldt-Jakob disease. This finding, however, does not support the use of the test in patients without clinically evident dementia. C1 NIH,CENT NERVOUS SYST STUDIES LAB,BASIC NEUROSCI PROGRAM,DIV INTRAMURAL RES,BETHESDA,MD 20892. CALTECH,DIV BIOL,PASADENA,CA 91125. RI Crozier, Laura/C-2170-2011 FU NIA NIH HHS [AG05131, P30AG1230]; NINDS NIH HHS [NS30531] NR 37 TC 470 Z9 481 U1 0 U2 4 PU MASS MEDICAL SOC PI BOSTON PA 10 SHATTUCK, BOSTON, MA 02115 SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD SEP 26 PY 1996 VL 335 IS 13 BP 924 EP 930 DI 10.1056/NEJM199609263351303 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA VK148 UT WOS:A1996VK14800003 PM 8782499 ER PT J AU Wichman, A AF Wichman, A TI Ethics of proxy consent for research involving patients with adult respiratory distress syndrome SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Letter RP Wichman, A (reprint author), NIH,BLDG 10,BETHESDA,MD 20892, USA. NR 6 TC 2 Z9 2 U1 0 U2 0 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD SEP 25 PY 1996 VL 276 IS 12 BP 949 EP 949 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA VH990 UT WOS:A1996VH99000019 PM 8805720 ER PT J AU White, L Petrovitch, H Ross, W Masaki, KH Abbott, RD Teng, EL Rodriguez, BL Blanchette, PL Havlik, RJ Wergowske, G Chiu, D Foley, DJ Murdaugh, C Curb, JD AF White, L Petrovitch, H Ross, W Masaki, KH Abbott, RD Teng, EL Rodriguez, BL Blanchette, PL Havlik, RJ Wergowske, G Chiu, D Foley, DJ Murdaugh, C Curb, JD TI Prevalence of dementia in older Japanese-American men in Hawaii - The Honolulu-Asia Aging Study SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID ALZHEIMERS-DISEASE; INFORMANT QUESTIONNAIRE; COGNITIVE DECLINE; SENILE DEMENTIA; ELDERLY IQCODE; POPULATION; EPIDEMIOLOGY; COMMUNITY; EDUCATION; STATE AB Objective.-To determine prevalence of dementia and its subtypes in Japanese-American men and compare these findings with rates reported for populations in Japan and elsewhere. Design and Setting.-The Honolulu Heart Program is a prospective population-based study of cardiovascular disease established in 1965. Prevalence estimates were computed from cases identified at the 1991 to 1993 examination. Cognitive performance was assessed using Standardized methods, instruments, and diagnostic criteria. Participants.-Subjects were 3734 Japanese-American men (80% of surviving cohort) aged 71 through 93 years, living in the community or in institutions. Main Outcome Measures.-Age-specific, age-standardized, and cohort prevalence estimates were computed for dementia (all cause) defined by 2 sets of diagnostic criteria and 4 levels of severity. Prevalence levels for Alzheimer disease and vascular dementia were also estimated. Results.-Dementia prevalence by Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised ranged from 2.1% in men aged 71 through 74 years to 33.4% in men aged 85 through 93 years. Age-standardized prevalence was 7.6%, Prevalence estimates for the cohort were 9.3% for dementia (all cause), 5.4% for Alzheimer disease (primary or contributing), and 4.2% for vascular dementia (primary or contributing). More than 1 possible cause was found in 26% of cases. The Alzheimer disease/vascular dementia ratio was 1.5 for cases attributed primarily to Alzheimer disease or vascular dementia. Conclusions.-Prevalence of Alzheimer disease in older Japanese-American men in Hawaii appears to be higher than in Japan but similar to European-ancestry populations. Prevalence of vascular dementia appears to be only slightly lower than in Japan, but higher than in European-ancestry populations. Further cross-national research with emphasis on standardized diagnostic methods is needed. C1 NIA,NIH,BETHESDA,MD 20892. NINR,NIH,BETHESDA,MD 20892. DEPT VET AFFAIRS,HONOLULU,HI. UNIV HAWAII,JOHN A BURNS SCH MED,DEPT MED,HONOLULU,HI 96822. UNIV VIRGINIA,DIV BIOSTAT,CHARLOTTESVILLE,VA. UNIV SO CALIF,LOS ANGELES,CA. RP White, L (reprint author), KUAKINI MED CTR,HONOLULU ASIA AGING STUDY,HONOLULU HEART PROGRAM,347 N KUAKININ ST,HONOLULU,HI 96817, USA. FU NHLBI NIH HHS [N0-HC-05102]; NIA NIH HHS [N01-AG-4-2149] NR 37 TC 307 Z9 307 U1 1 U2 7 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD SEP 25 PY 1996 VL 276 IS 12 BP 955 EP 960 DI 10.1001/jama.276.12.955 PG 6 WC Medicine, General & Internal SC General & Internal Medicine GA VH990 UT WOS:A1996VH99000035 PM 8805729 ER PT J AU Hoeg, JM AF Hoeg, JM TI Can genes prevent atherosclerosis? SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID HIGH-DENSITY-LIPOPROTEIN; CORONARY HEART-DISEASE; APOLIPOPROTEIN-A-I; ULTRAFAST COMPUTED-TOMOGRAPHY; FAMILIAL HYPERCHOLESTEROLEMIA; CHOLESTEROL LEVELS; TRANSGENIC MICE; FRAMINGHAM; MORTALITY; ATHEROGENESIS RP Hoeg, JM (reprint author), NHLBI,MOL DIS BRANCH,NIH,BLDG 10,ROOM 7N115,BETHESDA,MD 20892, USA. NR 48 TC 7 Z9 7 U1 0 U2 0 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD SEP 25 PY 1996 VL 276 IS 12 BP 989 EP 992 PG 4 WC Medicine, General & Internal SC General & Internal Medicine GA VH990 UT WOS:A1996VH99000041 PM 8805735 ER PT J AU Ford, RA Shaw, JA Cabib, E AF Ford, RA Shaw, JA Cabib, E TI Yeast chitin synthases 1 and 2 consist of a non-homologous and dispensable N-terminal region and of a homologous moiety essential for function SO MOLECULAR & GENERAL GENETICS LA English DT Article DE Saccharomyces cerevisiae; Chitin synthases; septum AB Predicted protein sequences of fungal chitin synthases can be divided into a non-homologous N-terminal region and a C-terminal region that shows significant homology among the various synthases. We have explored the function of these domains by constructing a series of nested deletions, extending from either end, in the CHS1 and CHS2 genes of Saccharomyces cerevisiae. In both cases, most or all of the sequences encoding the non-homologous N-terminal region (one-third of the protein for Chs1p and about one-fourth for Chs2p) could be excised, with little effect on the enzymatic activity in vitro of the corresponding synthase or on its function in vivo. However, further small deletions (20-25 amino acids) into the homologous region were deleterious to enzymatic activity and function., and often led to changes in the zymogenic character of the enzymes. Similarly, relatively small (about 75 amino acids) deletions from C-terminus resulted in loss of enzymatic activity and function of both synthases. Thus, it appears that all the information necessary for membrane localization, enzymatic activity and function resides in the homologous regions of Chs1p and Chs2p, a situation that may also apply to other chitin synthases. C1 NIDDKD, Lab Biochem & Metab, NIH, Bethesda, MD 20892 USA. RP Cabib, E (reprint author), NIDDKD, Lab Biochem & Metab, NIH, Bldg 10,Room 9N-115,10 Ctr Dr,MSC 1812, Bethesda, MD 20892 USA. NR 33 TC 19 Z9 19 U1 0 U2 0 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 SN 0026-8925 J9 MOL GEN GENET JI Mol. Gen. Genet. PD SEP 25 PY 1996 VL 252 IS 4 BP 420 EP 428 DI 10.1007/BF02173007 PG 9 WC Biochemistry & Molecular Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Genetics & Heredity GA VM601 UT WOS:A1996VM60100009 PM 8879243 ER PT J AU Quon, MJ Chen, H Lin, CH Zhou, LX Ing, BL Zarnowski, MJ Klinghoffer, R Kazlauskas, A Cushman, SW Taylor, SI AF Quon, MJ Chen, H Lin, CH Zhou, LX Ing, BL Zarnowski, MJ Klinghoffer, R Kazlauskas, A Cushman, SW Taylor, SI TI Effects of overexpressing wild-type and mutant PDGF receptors on translocation of GLUT4 in transfected rat adipose cells SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article ID PHOSPHATIDYLINOSITOL 3-KINASE ACTIVATION; GLUCOSE-TRANSPORT ACTIVITY; NEURONAL DIFFERENTIATION; INSULIN-RECEPTORS; PLASMA-MEMBRANE; PC12 CELLS; KINASE; SUBSTRATE-1; PROTEIN; SUBUNIT AB Activation of phosphatidylinositol 3-kinase (PI3K) by insulin is necessary for the effect of insulin to recruit GLUT4 to the cell surface in insulin target cells. In adipose cells, stimulation of endogenous PDGF receptors (PDGF-R) results in increased PI3K activity without causing recruitment of GLUT4. We overexpressed wild-type or mutant forms of the PDGF-R in rat adipose cells and examined their effects on PDGF- and insulin-stimulated recruitment of co-transfected epitope-tagged GLUT4. Control cells expressing only tagged GLUT4 had a 3-fold increase in cell surface GLUT4 upon insulin stimulation but no response to PDGF. Cells overexpressing wild-type PDGF-R maintained insulin responsiveness and, in addition, acquired the ability to recruit GLUT4 in response to PDGF. Surprisingly, overexpression of F740/ F751 (mutant PDGF-R unable to directly activate PI3K) led to similar results. Nevertheless, wortmannin (an inhibitor of PI3K) blocked effects of both PDGF and insulin to recruit GLUT4. Our data suggest that overexpression of PDGF-R mediates positive effects on GLUT4 translocation by a wortmannin sensitive pathway not dependent on direct interaction of the PDGF-R with PI3K. (C) 1996 Academic Press, Inc. C1 NIDDK,DIABET BRANCH,NIH,BETHESDA,MD 20892. NATL JEWISH CTR IMMUNOL & RESP MED,DENVER,CO 80206. RP Quon, MJ (reprint author), NHLBI,HYPERTENS ENDOCRINE BRANCH,NIH,BLDG 10,ROOM 8C-103,BETHESDA,MD 20892, USA. RI Quon, Michael/B-1970-2008; OI Quon, Michael/0000-0002-9601-9915; Quon , Michael /0000-0002-5289-3707 FU NIGMS NIH HHS [GM48339] NR 36 TC 26 Z9 26 U1 0 U2 1 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD SEP 24 PY 1996 VL 226 IS 3 BP 587 EP 594 DI 10.1006/bbrc.1996.1400 PG 8 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA VL164 UT WOS:A1996VL16400001 PM 8831662 ER PT J AU Dustan, HP Roccella, EJ Garrison, HH AF Dustan, HP Roccella, EJ Garrison, HH TI Controlling hypertension - A research success story SO ARCHIVES OF INTERNAL MEDICINE LA English DT Article ID PREVALENCE; POPULATION; HEALTH AB In the past 2 decades, deaths from stroke have decreased by 59% and deaths from heart attack by 53%. An important component of this dramatic change has been the increased use of antihypertensive drugs. This remarkable success resulted from broad-based and diverse research programs supported by the federal government, pharmaceutical companies, voluntary health agencies, and private foundations. It included basic research, drug development programs, epidemiologic studies, health surveys of US citizens, clinical research, and large-scale drug trials. Four of the categories of antihypertensive drugs in wide use-diuretics, beta-blockers, calcium antagonists, and angiotensin-converting ensyme inhibitors-emerged from widely different areas of investigation. In the beginning, dir major breakthroughs that led to the development of these drugs were impossible to forecast, and their ultimate applications were impossible to predict. Although decreases in hypertension-related mortality are impressive, enthusiasm must be tempered because the mechanisms of hypertension are still incompletely understood and, prevention is not yet possible. Continued research is needed to extend these advances. C1 FED AMER SOC EXPT BIOL,OFF PUBL AFFAIRS,BETHESDA,MD 20814. UNIV VERMONT,COLL MED,DEPT PHARMACOL,BURLINGTON,VT 05405. UNIV VERMONT,COLL MED,DEPT MED,BURLINGTON,VT 05405. NHLBI,NATL HIGH BLOOD PRESSURE EDUC PROGRAM,BETHESDA,MD 20892. OI Garrison, Howard/0000-0001-9455-3527 NR 65 TC 25 Z9 25 U1 0 U2 0 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 SN 0003-9926 J9 ARCH INTERN MED JI Arch. Intern. Med. PD SEP 23 PY 1996 VL 156 IS 17 BP 1926 EP 1935 DI 10.1001/archinte.156.17.1926 PG 10 WC Medicine, General & Internal SC General & Internal Medicine GA VJ676 UT WOS:A1996VJ67600005 PM 8823146 ER PT J AU Porush, JG August, PA Bakris, GL Brenner, BM Levine, SA Buckalew, VM Flack, JM Klag, MJ Vidt, DG Roccella, EJ Anderson, DE AF Porush, JG August, PA Bakris, GL Brenner, BM Levine, SA Buckalew, VM Flack, JM Klag, MJ Vidt, DG Roccella, EJ Anderson, DE TI 1995 update of the working group reports on chronic renal failure and renovascular hypertension - National High Blood Pressure Education Program Working Group SO ARCHIVES OF INTERNAL MEDICINE LA English DT Article ID CONVERTING ENZYME-INHIBITION; URINARY ALBUMIN EXCRETION; FACTOR INTERVENTION TRIAL; II DIABETIC-PATIENTS; ANTIHYPERTENSIVE TREATMENT; ARTERY STENOSIS; ABDOMINAL-AORTA; NATURAL-HISTORY; MINORITY-GROUPS; MR ANGIOGRAPHY AB To update 2 National High Blood Pressure Education Program working group reports on hypertension and chronic renal failure and renovascular hypertension, a working group was appointed by the director of the National Heart, Lung, and Blood Institute. Literature was searched through MEDLINE and the National Heart, Lung, and Blood Institute information center library. Scientific evidence was given precedence over clinical anecdotal experience. The working group members produced initial draft documents that were circulated to additional experts on hypertension and renal disease. This reiterative process occurred for 18 draft documents. The final report was sent to the representatives of the 44 organizations on the Coordinating Committee for vote and unanimously approved September 1, 1995. The report recommended treatment of hypertension to the goal of 130/85 mm Hg with whatever therapy is necessary to prevent the development of hypertensive nephrosclerosis or the progression of established renal disease of diverse causes. It seems reasonable to recommend angiotensin-converting enzyme inhibitors as initial therapy for patients with diabetes and microalbuminuria or overt diabetic nephropathy with and without hypertension. Renovascular disease has emerged as a major cause of end-stage renal disease, especially in the elderly. Newer screening procedures for the noninvasive screening of renovascular disease include the captopril test, renal scintigraphy following captopril administration, duplex scanning, and magnetic resonance angiography. C1 NHLBI, NIH, NATL HIGH BLOOD PRESSURE EDUC PROGRAM, BETHESDA, MD 20892 USA. SUNY HLTH SCI CTR, BROOKDALE HOSP MED CTR, DIV NEPHROL & HYPERTENS, BROOKLYN, NY USA. NEW YORK HOSP, CTR CARDIOVASC, NEW YORK, NY 10021 USA. RUSH PRESBYTERIAN ST LUKES MED CTR, HYPERTENS FELLOWSHIP PROGRAM, CHICAGO, IL 60612 USA. BRIGHAM & WOMENS HOSP, DIV RENAL, BOSTON, MA 02115 USA. WAKE FOREST UNIV, BOWMAN GRAY SCH MED, HYPERTENS CTR, WINSTON SALEM, NC USA. JOHNS HOPKINS MED INST, WELCH CTR PREVENT EPIDEMIOL & CLIN RES, BALTIMORE, MD 21205 USA. CLEVELAND CLIN FDN, DEPT HYPERTENS & NEPHROL, CLEVELAND, OH 44195 USA. ROW SCI INC, NATL HIGH BLOOD PRESSURE EDUC PROGRAM, ROCKVILLE, MD USA. NIDDK, NIH, BETHESDA, MD USA. NYU, MED CTR, RENAL SECT, NEW YORK, NY 10016 USA. UNIV COLORADO, COLL PHARM, DEPT PHARM PRACTICE, DENVER, CO 80202 USA. NHLBI, NIH, DIV EPIDEMIOL & CLIN APPLICAT, CLIN APPLICAT & PREVENT PROGRAMS, BETHESDA, MD USA. CASE WESTERN RESERVE UNIV, UNIV HOSP CLEVELAND, DIV HYPERTENS, CLEVELAND, OH 44106 USA. VET ADM MED CTR, SERV NEPHROL, MIAMI, FL 33125 USA. UNIV KANSAS, MED CTR, DIV NEPHROL & HYPERTENS, KANSAS CITY, KS 66103 USA. VET ADM MED CTR, NEPHROL SECT, LONG BEACH, CA 90822 USA. UNIV CINCINNATI, MED CTR, DEPT MED, CINCINNATI, OH 45267 USA. BOWMAN GRAY SCH MED, UROL CLIN, DANVILLE, VA USA. UNIV MARYLAND, DIV HYPERTENS, BALTIMORE, MD 21201 USA. WAYNE STATE UNIV, DIV ENDOCRINOL METAB & HYPERTENS, DETROIT, MI USA. MAYO CLIN & MAYO FDN, MAYO MED SCH, DIV HYPERTENS & INTERNAL MED, ROCHESTER, MN 55905 USA. CASE WESTERN RESERVE UNIV, UNIV HOSP CLEVELAND, CLIN HYPERTENS PROGRAM, CLEVELAND, OH 44106 USA. VET AFFAIRS MED CTR, HYPERTENS SECT, CLEVELAND, OH USA. VET AFFAIRS MED CTR, HYPERTENS LIPID CLIN, CLEVELAND, OH USA. NR 98 TC 73 Z9 75 U1 1 U2 2 PU AMER MEDICAL ASSOC PI CHICAGO PA 330 N WABASH AVE, STE 39300, CHICAGO, IL 60611-5885 USA SN 0003-9926 EI 1538-3679 J9 ARCH INTERN MED JI Arch. Intern. Med. PD SEP 23 PY 1996 VL 156 IS 17 BP 1938 EP 1947 PG 10 WC Medicine, General & Internal SC General & Internal Medicine GA VJ676 UT WOS:A1996VJ67600006 ER PT J AU Davis, CD Dacquel, EJ Schut, HAJ Thorgeirsson, SS Snyderwine, EG AF Davis, CD Dacquel, EJ Schut, HAJ Thorgeirsson, SS Snyderwine, EG TI In vivo mutagenicity and DNA adduct levels of heterocyclic amines in Muta(TM) Mice and c-myc/lacZ double transgenic mice SO MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS LA English DT Article DE 2-amino-3-methylimidazo[4,5-f]quinoline; 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline; 2-amino-9H-pyrido[2,3-b]indole; P-32-postlabeling; in vivo mutagenicity ID CYNOMOLGUS MONKEYS; ESCHERICHIA-COLI; IN-VIVO; LIVER; GENE; FOOD; RAT; 2-AMINO-3-METHYLIMIDAZO<4,5-F>QUINOLINE; ACTIVATION; IQ AB The cooked meat derived heterocyclic amines (HCAs) 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), and 2-amino-9H-pyrido[2,3-b]indole (A alpha C) are established mutagens in the Salmonella assay and hepatocarcinogens in mice. The current study uses transgenic mice to examine hepatic HCA-DNA adduct formation and mutagenesis in vivo and the impact of hepatic overexpression of the c-myc oncogene on HCA-induced mutagenesis. C57B1/lacZ and c-myc/lacZ mice strains, produced by crossbreeding Muta(TM)Mice (carrying the lacZ mutation target gene) with either C57B1 control or c-myc transgenic mice, respectively, were treated with 10 daily doses of IQ, MeIQx or A alpha C (20 mu g/g, p.o.). Four weeks after dosing, the frequency of mutations in the lacZ gene in liver of either C57B1/lacZ or c-myc/lacZ mice was significantly higher in mice treated with any one of the three HCAs than in mice given vehicle only. In addition, all three HCAs formed hepatic DNA adducts, as measured by the P-32-postlabeling analysis 24 h after dosing. In both strains of mice, hepatic DNA adduct levels were 2-3-fold higher with A alpha C than with either IQ or MeIQx, although the mutant frequencies in the lacZ gene were 30-40% lower in mice dosed with A alpha C. These results suggest that A alpha C-DNA adducts may be less mutagenic in vivo than either IQ- or MeIQx-DNA adducts. The lacZ mutant frequencies observed with all three HCAs appeared to be influenced by c-myc transgene expression: after HCA treatment, transgenic mice carrying the c-myc gene showed a 30-40% higher lacZ mutant frequency than mice not carrying this transgene. Notably, lacZ mutant frequencies were not different among C57B1/lacZ and c-myc/lacZ mice that received vehicle control. DNA adduct studies showed that the levels of IQ- and MeIQx-DNA adducts were 2-3-fold higher in c-myc/lacZ mice than in C57B1/lacZ mice; however, A alpha C-DNA adducts were not statistically different between the two strains. In addition, phase I metabolic activation of these HCAs, as assessed by hepatic microsomal mutagenic activation, was also similar in both strains of mice. These results support the notion that overexpression of the c-myc oncogene cooperates with the HCAs to enhance in vivo mutagenicity. Further studies are needed to assess the mechanisms of this cooperative effect. C1 NCI,EXPT CARCINOGENESIS LAB,BETHESDA,MD 20892. MED COLL OHIO,DEPT PATHOL,TOLEDO,OH 43614. NR 32 TC 31 Z9 31 U1 1 U2 3 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 SEP 23 PY 1996 VL 356 IS 2 BP 287 EP 296 DI 10.1016/0027-5107(96)00074-7 PG 10 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology SC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology GA VJ559 UT WOS:A1996VJ55900022 PM 8841498 ER PT J AU Mascola, JR Louder, MK Surman, SR Vancott, TC Yu, XF Bradac, J Porter, KR Nelson, KE Girard, M McNeil, JG McCutchan, FE Birx, DL Burke, DS AF Mascola, JR Louder, MK Surman, SR Vancott, TC Yu, XF Bradac, J Porter, KR Nelson, KE Girard, M McNeil, JG McCutchan, FE Birx, DL Burke, DS TI Human immunodeficiency virus type 1 neutralizing antibody serotyping using serum pools and an infectivity reduction assay SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article ID MONOCLONAL-ANTIBODIES; ENVELOPE GLYCOPROTEIN; PHYLOGENETIC ANALYSIS; NORTHERN THAILAND; HIV TYPE-1; DIVERSITY; SUBTYPES; GP120; AIDS; SENSITIVITY AB Classification of human immunodeficiency virus type 1 (HIV-1) by neutralization serotype may be important for the design of active and passive immunization strategies. Neutralizing antibody serotyping is hindered by the lack of standard reagents and assay format, and by the weak activity of many individual sera. To facilitate cross-clade neutralization analysis, we used an infectivity reduction assay (IRA) and selected clade-specific serum (or plasma) pools from subjects infected with clade B and E HIV-1, respectively, Several serum pools were utilized; some were selected for strong neutralizing activity against intraclade viruses and others were derived from conveniently available samples, Against a panel of 51 clade B and E viruses, serum pools displayed strong neutralization of most intraclade viruses and significantly diminished cross-clade neutralization, Results mere confirmed against a blinded panel of 20 viruses, The data indicate that the phylogenetic classification of virus subtypes B and E corresponds to two distinct neutralization serotypes, This approach to neutralizing antibody serotyping may be useful in defining the antigenic relationship among viruses from other clades. C1 USN,MED RES INST,DEPT INFECT DIS,BETHESDA,MD 20889. HENRY M JACKSON FDN,ROCKVILLE,MD 20850. JOHNS HOPKINS UNIV,SCH HYG & PUBL HLTH,BALTIMORE,MD 21205. NIAID,DIV AIDS,NIH,ROCKVILLE,MD 20892. INST PASTEUR,PARIS,FRANCE. RP Mascola, JR (reprint author), WALTER REED ARMY INST RES,DIV RETROVIROL,13 TAFT COURT,SUITE 200,ROCKVILLE,MD 20850, USA. OI /0000-0002-5704-8094 FU NIDA NIH HHS [DA 09541] NR 43 TC 67 Z9 68 U1 0 U2 0 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 SN 0889-2229 J9 AIDS RES HUM RETROV JI Aids Res. Hum. Retrovir. PD SEP 20 PY 1996 VL 12 IS 14 BP 1319 EP 1328 DI 10.1089/aid.1996.12.1319 PG 10 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA VJ248 UT WOS:A1996VJ24800004 PM 8891111 ER PT J AU Murthy, KK Cobb, EK ElAmad, Z Ortega, H Hsueh, FC Satterfield, W Lee, DR Kalish, ML Haigwood, NL Kennedy, RC Steimer, KS Schultz, A Levy, JA AF Murthy, KK Cobb, EK ElAmad, Z Ortega, H Hsueh, FC Satterfield, W Lee, DR Kalish, ML Haigwood, NL Kennedy, RC Steimer, KS Schultz, A Levy, JA TI Titration of a vaccine stock preparation of Human Immunodeficiency Virus type 1(SF2) in cultured lymphocytes and in chimpanzees SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article ID LYMPHADENOPATHY-ASSOCIATED VIRUS; POLYMERASE CHAIN-REACTION; ANTI-HIV ACTIVITY; PERSISTENT INFECTION; GLYCOPROTEIN GP120; CHALLENGE; PROTECTION; PREVENTION; CELLS; AIDS AB A large stock preparation of the HIV-1(SF2) isolate has been derived after serial passage in human peripheral blood mononuclear cells (PBMCs). This viral stock has a titer of 10(4.9) TCID50 in human PBMCs and 10(4.2) TCID50 in chimpanzee PBMCs, Bg inoculation into animals the 50% chimpanzee infectious dose titer was found to be about 10(2.3). Virus isolation from animals was achieved on most occasions within 1-4 weeks after inoculation and then became transient, Viral RNA and DNA PCR analyses confirmed the virus infection of the chimpanzees, Anti-HIV antibody levels in the inoculated animals ranged from 1:400 to 1:6400 as measured by ELISA, About 680 vials of this stock preparation, frozen at -190 degrees C, are available for future studies of vaccines and antiviral therapies. C1 CHIRON CORP,EMERYVILLE,CA 94608. UNIV CALIF SAN FRANCISCO,CANC RES INST,SAN FRANCISCO,CA 94143. UNIV TEXAS,MD ANDERSON CANC CTR,DEPT VET SCI,BASTROP,TX 78602. CTR DIS CONTROL,DIV HIV AIDS,ATLANTA,GA 30333. BRISTOL MYERS SQUIBB CO,SEATTLE,WA 98121. UNIV OKLAHOMA,HLTH SCI CTR,DEPT MICROBIOL & IMMUNOL,OKLAHOMA CITY,OK 73190. NIAID,VACCINE BRANCH,BETHESDA,MD 20892. UNIV CALIF SAN FRANCISCO,DEPT MED,SAN FRANCISCO,CA 94143. RP Murthy, KK (reprint author), SW FDN BIOMED RES,DEPT VIROL & IMMUNOL,POB 760549,SAN ANTONIO,TX 78245, USA. FU NIAID NIH HHS [AI-24286, AI-05060, AI-26462] NR 37 TC 16 Z9 16 U1 0 U2 0 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 SN 0889-2229 J9 AIDS RES HUM RETROV JI Aids Res. Hum. Retrovir. PD SEP 20 PY 1996 VL 12 IS 14 BP 1341 EP 1348 DI 10.1089/aid.1996.12.1341 PG 8 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA VJ248 UT WOS:A1996VJ24800006 PM 8891113 ER PT J AU Robbins, KE Bandea, CI Levin, A Goedert, JJ Blattner, WA Brubaker, G Brown, TM Schochetman, G Kalish, ML Shao, J OBrien, TR AF Robbins, KE Bandea, CI Levin, A Goedert, JJ Blattner, WA Brubaker, G Brown, TM Schochetman, G Kalish, ML Shao, J OBrien, TR TI Genetic variability of human immunodeficiency virus type 1 in rural northwest Tanzania SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article ID DIVERSITY C1 ST BARTHOLOMEWS HOSP,RES TRIANGLE INST UNIT,LONDON EC1M 6AH,ENGLAND. SHIRATI HOSP,SHIRATI,TANZANIA. NCI,VIRAL EPIDEMIOL BRANCH,PUBL HLTH SERV,US DEPT HHS,ROCKVILLE,MD 20852. CTR DIS CONTROL & PREVENT,DIV AIDS SEXUALLY TRANSMITTED DIS & TB LAB RES,NATL CTR INFECT,ATLANTA,GA 30333. FU NCI NIH HHS [NIC-CP-EB-85603-57, N0I-CP-33005] NR 11 TC 17 Z9 17 U1 0 U2 0 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 SN 0889-2229 J9 AIDS RES HUM RETROV JI Aids Res. Hum. Retrovir. PD SEP 20 PY 1996 VL 12 IS 14 BP 1389 EP 1391 DI 10.1089/aid.1996.12.1389 PG 3 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA VJ248 UT WOS:A1996VJ24800013 PM 8891120 ER PT J AU Blacker, D Faraone, SV Rosen, AE Guroff, JJ Adams, P Weissman, MM Gershon, ES AF Blacker, D Faraone, SV Rosen, AE Guroff, JJ Adams, P Weissman, MM Gershon, ES TI Unipolar relatives in bipolar pedigrees: A search for elusive indicators of underlying bipolarity SO AMERICAN JOURNAL OF MEDICAL GENETICS LA English DT Article DE bipolar disorder; major depression; linkage analysis; caseness index; phenotype ID MANIC-DEPRESSIVE ILLNESS; OBSESSIVE-COMPULSIVE DISORDER; LINKAGE ANALYSIS; GENETIC-LINKAGE; PSYCHIATRIC-DISORDERS; MAJOR DEPRESSION; CLINICAL CHARACTERISTICS; COLLABORATIVE PROGRAM; COMPLEX DISEASES; DNA MARKERS AB In an effort to identify features indicative of underlying bipolarity within the unipolar relatives of bipolar probands, we compared unipolar relatives of bipolars with unipolar relatives of controls. Using data from the Yale-NIMH Collaborative Study of Depression, we compared a number of demographic and clinical features individually, and then developed a logistic regression model for the differences found. Unipolar relatives of bipolars were generally similar to relatives of controls, but they were older and more likely to suffer from more severe, even psychotic, depression, and somewhat less likely to report a brief transition into their illness. A multiple logistic regression model for observed differences was highly statistically significant, but had limited ability to discriminate effectively between the two groups. These findings suggest that more stringent diagnostic criteria might be beneficial if unipolar relatives are counted as affected in linkage studies of bipolar disorder. The ability of this strategy to improve the ''clinical phenotype'' is limited, however, and other approaches may be needed to identify features of underlying bipolarity and thus to define ''caseness'' for unipolar relatives in linkage analyses of bipolar disorder. (C) 1996 Wiley-Liss, Inc. C1 HARVARD UNIV,SCH MED,DEPT PSYCHIAT,BOSTON,MA. MASSACHUSETTS GEN HOSP,BOSTON,MA 02114. MASSACHUSETTS MENTAL HLTH CTR,BOSTON,MA 02115. BROCKTON W ROXBURY VET AFFAIRS MED CTR,BROCKTON,MA. HARVARD UNIV,SCH MED,INST PSYCHIAT EPIDEMIOL & GENET,BOSTON,MA. HARVARD UNIV,SCH PUBL HLTH,BOSTON,MA 02115. COLUMBIA UNIV,COLL PHYS & SURG,DEPT PSYCHIAT,NEW YORK,NY. NEW YORK PSYCHIAT INST,DIV CLIN GENET EPIDEMIOL,NEW YORK,NY. NIMH,CLIN NEUROGENET BRANCH,BETHESDA,MD 20892. OI Weissman, Myrna/0000-0003-3490-3075; Faraone, Stephen/0000-0002-9217-3982 FU NIMH NIH HHS [K21-MH01118, R01-MH28274] NR 68 TC 15 Z9 15 U1 2 U2 3 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC 605 THIRD AVE, NEW YORK, NY 10158-0012 SN 0148-7299 J9 AM J MED GENET JI Am. J. Med. Genet. PD SEP 20 PY 1996 VL 67 IS 5 BP 445 EP 454 DI 10.1002/(SICI)1096-8628(19960920)67:5<445::AID-AJMG2>3.0.CO;2-J PG 10 WC Genetics & Heredity SC Genetics & Heredity GA VJ223 UT WOS:A1996VJ22300002 PM 8886160 ER PT J AU Osorio, M Torres, J Moya, F Pezzullo, J Salafia, C Baxter, R Schwander, J Fant, M AF Osorio, M Torres, J Moya, F Pezzullo, J Salafia, C Baxter, R Schwander, J Fant, M TI Insulin-like growth factors (IGFs) and IGF binding proteins-1, -2, and -3 in newborn serum: Relationships to fetoplacental growth at term SO EARLY HUMAN DEVELOPMENT LA English DT Article DE IGFs; IGF binding proteins; fetal growth; placental growth ID FACTOR-II; INTRAUTERINE GROWTH; CIRCULATING LEVELS; HUMAN-FETUS; CORD BLOOD; SOMATOMEDIN; TISSUES; RAT AB Cord sera were obtained from term, Chilean newborns exhibiting various patterns of intrauterine growth and assayed for IGF-1, IGF-2, IGFBP-1, IGFBP-2, and IGFBP-3 by specific radioimmunoassays (RIA). Serum levels of each peptide were correlated with birth weight (BW), ponderal index (PI), and placental weight (PW). Total IGF-1 levels correlated with BW (r = 0.665, P = 0.0001), PI (r = 0.527, P = 0.004), and PW (r = 0.596, P = 0.0017). In contrast, IGF-2 failed to correlate with any growth parameter. Of the three binding proteins, IGFBP-3 exhibited the strongest relationship to each growth parameter. IGFBP-3 correlated significantly with BW (r = 0.71, P < 0.0001), PI (r = 0.782, P < 0.0001), and PW (r = 0.57, P = 0.0029). In addition IGFBP-3 levels positively correlated to IGF-I levels (r = 0.614, P = 0.0005). By contrast, circulating IGFBP-1 and IGFBP-2 were inversely related to IGF-1 levels. All five peptides were subjected to multiple regression analysis and related to BW. Significant relationships between the predicted BW and the actual BW were observed in these infants (r = 0.802, P = 0.0006). The BWs of a cohort of unrelated North American infants were also predicted using the Chilean-derived equation and found to be significantly related to their actual BWs (r = 0.453, P = 0.0033). These relationships were strengthened by the inclusion of estimated gestational age (EGA) as an independent variable. These data point to particularly important roles for IGF-1 and IGFBP-3 in regulating fetal growth at term, and suggest that they are regulated in a coordinated manner during the latter stage of gestation. Furthermore, they suggest that IGFBPs play multiple, and potentially opposing, regulatory roles in modulating IGF action. Lastly, an integrated expression of IGF activity derived from one population significantly correlated with newborn BW in a geographically and culturally distinct population. C1 GEORGETOWN UNIV,SCH MED,DEPT OBSTET & GYNECOL,WASHINGTON,DC 20007. UNIV TEXAS,SW MED CTR,DALLAS,TX 75235. UNIV CHILE,FAC MED,DEPT PEDIAT,SANTIAGO 7,CHILE. GEORGETOWN UNIV,SCH MED,DEPT PATHOL,WASHINGTON,DC. NICHHD,PERINATOL RES BRANCH,BETHESDA,MD 20892. ROYAL N SHORE HOSP,KOLLING INST MED RES,SYDNEY,NSW,AUSTRALIA. KANTONSSPITAL,DEPT MED,BASEL,SWITZERLAND. WASHINGTON UNIV,SCH MED,ST LOUIS CHILDRENS HOSP,EDWARD G MALLINKRODT DEPT PEDIAT,ST LOUIS,MO 63110. FU NICHD NIH HHS [HD22993] NR 23 TC 51 Z9 52 U1 0 U2 1 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0378-3782 J9 EARLY HUM DEV JI Early Hum. Dev. PD SEP 20 PY 1996 VL 46 IS 1-2 BP 15 EP 26 DI 10.1016/0378-3782(96)01737-9 PG 12 WC Obstetrics & Gynecology; Pediatrics SC Obstetrics & Gynecology; Pediatrics GA VN650 UT WOS:A1996VN65000003 PM 8899351 ER PT J AU GopalSrivastava, R Cvekl, A Piatigorsky, J AF GopalSrivastava, R Cvekl, A Piatigorsky, J TI Pax-6 and alpha B-crystallin/small heat shock protein gene regulation in the murine lens - Interaction with the lens-specific regions, LSR1 and LSR2 SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HOMEOBOX-CONTAINING GENE; RETINOIC ACID RECEPTOR; PAIRED DOMAIN; EYELESS GENE; EXPRESSION; IDENTIFICATION; DNA; FAMILY; MUTATIONS; ENHANCER AB We have demonstrated previously that a transgene comprising the -164/+44 fragment of the murine alpha B-crystallin gene fused to the bacterial chloramphenicol acetyltransferase (cat) gene is lens-specific in transgenic mice. The -147 to -118 sequence was identified as a lens-specific regulatory region and is called here LSR1 for lens-specific region 1. In the present experiments, a -115/+44-cat transgene was also lens specific in transgenic mice, although the average activity was 30 times lower than that derived from the -164/+44-cat transgene. The -115/+44 alpha B-crystallin fragment contains a highly conserved region (-78 to -46) termed here LSR2. A -68/+44-cat transgene, in which LSR2 is truncated, was inactive in transgenic mice. DNase I footprinting indicated that LSR1 and LSR2 bind partially purified nuclear proteins from either alpha TN4-1 lens cells or the mouse lens as well as the purified paired domain of Pax-6. Site-specific mutation of LSR1 eliminated both Pax-6 binding and promoter activity of the -164/+44-cat transgene in transgenic mice. Finally antibody/electrophoretic mobility shift assays and cotransfection experiments indicated that Pax-6 can activate the alpha B-crystallin promoter via LSR1 and LSR2. Our data strengthen the idea that Pax-6 has had a major role in recruiting genes for high expression in the lens. C1 NEI, MOL & DEV BIOL LAB, NIH, BETHESDA, MD 20892 USA. RI Cvekl, Ales/B-2427-2013 NR 52 TC 57 Z9 57 U1 0 U2 2 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 SEP 20 PY 1996 VL 271 IS 38 BP 23029 EP 23036 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VH768 UT WOS:A1996VH76800017 PM 8798491 ER PT J AU Sutton, J Costa, R Klug, M Field, L Xu, DW Largaespada, DA Fletcher, CF Jenkins, NA Copeland, NG Klemsz, M Hromas, R AF Sutton, J Costa, R Klug, M Field, L Xu, DW Largaespada, DA Fletcher, CF Jenkins, NA Copeland, NG Klemsz, M Hromas, R TI Genesis, a winged helix transcriptional repressor with expression restricted to embryonic stem cells SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID FORK HEAD DOMAIN; HEPATOCYTE NUCLEAR FACTOR-3-BETA; DNA-BINDING; ALVEOLAR RHABDOMYOSARCOMA; REGULATORY ELEMENTS; MAMMALIAN-CELLS; FUSION PROTEIN; FAMILY; MOUSE; CLONING AB A novel member of the winged helix (formerly HNF-3/Forkhead) transcriptional regulatory family, termed Genesis, was isolated and characterized. Putative translation of the complete cDNA revealed the winged helix DNA binding domain to be centrally located within the protein, with regions on either side that contain known transcriptional regulatory motifs. Extensive Northern analysis of Genesis found that the message was exclusively expressed in embryonic stem cells or their malignant equivalent, embryonal carcinoma cells. The Genesis transcript was down-regulated when these cells were stimulated to differentiate. DNA sequences that Genesis protein would interact with were characterized and were found to contain a consensus similar to that found in an embryonic stem cell enhancer sequence, Go-transfection experiments revealed that Genesis is a transcriptional repressor. Genesis mapped to mouse chromosome 4 in a region syntenic with human chromosome 1p31, a site of nonrandom abnormalities in germ cell neoplasia, neuroblastoma, and acute lymphoblastic leukemia, Genesis is a candidate for regulating the phenotype of normal or malignant embryonic stem cells. C1 INDIANA UNIV,MED CTR,DIV HEMATOL ONCOL,INDIANAPOLIS,IN 46202. INDIANA UNIV,MED CTR,WALTHER ONCOL CTR,INDIANAPOLIS,IN 46202. UNIV ILLINOIS,COLL MED,DEPT BIOCHEM,CHICAGO,IL 60612. INDIANA UNIV,MED CTR,KRANNERT INST CARDIOL,INDIANAPOLIS,IN 46202. NCI,MAMMALIAN GENET LAB,ABL BASIC RES PROGRAM,NIH,FREDERICK CANC RES & DEV CTR,FREDERICK,MD 21702. INDIANA UNIV,MED CTR,DEPT MICROBIOL & IMMUNOL,INDIANAPOLIS,IN 46202. RI Largaespada, David/C-9832-2014 FU NHLBI NIH HHS [HL 48915]; NIGMS NIH HHS [GM43241] NR 65 TC 145 Z9 151 U1 0 U2 3 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD SEP 20 PY 1996 VL 271 IS 38 BP 23126 EP 23133 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VH768 UT WOS:A1996VH76800031 PM 8798505 ER PT J AU Jideama, NM Noland, TA Raynor, RL Blobe, GC Fabbro, D Kazanietz, MG Blumberg, PM Hannun, YA Kuo, JF AF Jideama, NM Noland, TA Raynor, RL Blobe, GC Fabbro, D Kazanietz, MG Blumberg, PM Hannun, YA Kuo, JF TI Phosphorylation specificities of protein kinase C isozymes for bovine cardiac troponin I and troponin T and sites within these proteins and regulation of myofilament properties SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ACTOMYOSIN MGATPASE ACTIVITY; MUSCLE-CONTRACTION; CA2+-STIMULATED MGATPASE; RECONSTITUTED ACTOMYOSIN; HEAVY-MEROMYOSIN; MYOCARDIAL-CELLS; RAT-HEART; ACTIN; TROPOMYOSIN; EXPRESSION AB Protein kinase C (PKC) isozymes alpha, delta, epsilon, and zeta, shown to be expressed in adult rat cardiomyocytes, displayed distinct substrate specificities in phosphorylating troponin I and troponin T subunits in the bovine cardiac troponin complex, Thus, because they have different substrate affinities, PKC-alpha, -delta, and -epsilon phosphorylated troponin I more than troponin T, but PKC-zeta conversely phosphorylated the latter more than the former, Furthermore, PKC isozymes exhibited discrete specificities in phosphorylating distinct sites in these proteins as free subunits or in the troponin complex, Unlike other isozymes, PKC-delta was uniquely able to phosphorylate Ser-23/Ser-24 in troponin I, the bona fide phosphorylation sites for protein kinase A (PKA); and consequently, like PKA, it reduced Ca2+ sensitivity of Ca2+-stimulated MgATPase of reconstituted actomyosin S-1. In addition, PKC-delta, like PKC-alpha, readily phosphorylated Ser-43/Ser-45 (sites common for all PKC isozymes) and reduced maximal activity of MgATPase, In this respect, PKC-delta functioned as a hybrid of PKC-alpha and PKA. In contrast to PKC-alpha, -delta, and -epsilon, PKC-zeta exclusively phosphorylated two previously unknown sites in troponin T. Phosphorylation of troponin T by PKC-alpha resulted in decreases in both Ca2+ sensitivity and maximal activity, whereas phosphorylation by PKC-zeta resulted in a slight increase of the Ca2+ sensitivity without affecting the maximal activity of MgATPase, Most of the in vitro phosphorylation sites in troponin I and troponin T were confirmed in situ in adult rat cardiomyocytes. The present study has demonstrated for the first time distinct specificities of PKC isozymes for phosphorylation of two physiological substrates in the myocardium, with functional consequences. C1 EMORY UNIV, SCH MED, DEPT PHARMACOL, ATLANTA, GA 30322 USA. DUKE UNIV, MED CTR, DEPT MED HEMATOL ONCOL, DURHAM, NC 27710 USA. DUKE UNIV, MED CTR, DEPT CELL BIOL, DURHAM, NC 27710 USA. CIBA GEIGY LTD, DEPT PHARMACEUT RES, ONCOL K125, CH-4002 BASEL, SWITZERLAND. NCI, MOL MECHANISMS TUMOR PROMOT SECT, CELLULAR CARCINOGENESIS & TUMOR PROMOT LAB, NIH, BETHESDA, MD 20892 USA. FU NHLBI NIH HHS [HL-15696, HL-43707] NR 42 TC 142 Z9 143 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 EI 1083-351X J9 J BIOL CHEM JI J. Biol. Chem. PD SEP 20 PY 1996 VL 271 IS 38 BP 23277 EP 23283 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VH768 UT WOS:A1996VH76800052 PM 8798526 ER PT J AU Adler, V Pincus, MR Polotskaya, A Montano, X Friedman, FK Ronai, Z AF Adler, V Pincus, MR Polotskaya, A Montano, X Friedman, FK Ronai, Z TI Activation of c-Jun-NH2-kinase by UV irradiation is dependent on p21(ras) SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID C-JUN KINASE; PROTEIN-KINASE; IONIZING-RADIATION; AP-1 ACTIVITY; CELLS; RAS; EXPRESSION; PHOSPHORYLATION; INDUCTION; PATHWAYS AB We have demonstrated previously that Jun-NH2-kinase (JNK) activation in vitro is potentiated by association with the p21(ras) protein. To determine if in vivo activation of JNK also depends on p21(ras), we have used M1311 cells that carry the cDNA for the neutralizing antibody to p21(ras), Y13-259, under a dexamethasone-inducible promoter. The ability of UV to activate JNK gradually decreased over a 4-day period of cell growth in dexamethasone. This decrease coincides with weaker transcriptional activation measured via gel shift and chloramphenicol acetyltransferase assays. Peptides corresponding to amino acids 96-110 on p21(ras), which were shown to block Ras-JNK association, inhibited UV-mediated JNK activation in mouse fibroblast 3T3-4A cells as well as in M1311 cells, further supporting the role of p21(ras) in UV-mediated JNK activation. Overall, the present studies provide in vivo confirmation of the role p21(ras) plays in JNK activation by UV irradiation. C1 AMER HLTH FDN,MOL CARCINOGENESIS PROGRAM,VALHALLA,NY 10595. VET AFFAIRS MED CTR,DEPT PATHOL & LAB MED,BROOKLYN,NY 11209. SUNY HLTH SCI CTR,BROOKLYN,NY 11203. IMPERIAL CANC RES FUND,LONDON WC2A 3PX,ENGLAND. NCI,MOL CARCINOGENESIS LAB,NIH,BETHESDA,MD 20892. RI Friedman, Fred/D-4208-2016; OI RONAI, ZEEV/0000-0002-3859-0400 FU NCI NIH HHS [CA51995, CA42500] NR 30 TC 74 Z9 74 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD SEP 20 PY 1996 VL 271 IS 38 BP 23304 EP 23309 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VH768 UT WOS:A1996VH76800056 PM 8798530 ER PT J AU Miller, MJ Martinez, A Unsworth, EJ Thiele, CJ Moody, TW Elsasser, T Cuttitta, F AF Miller, MJ Martinez, A Unsworth, EJ Thiele, CJ Moody, TW Elsasser, T Cuttitta, F TI Adrenomedullin expression in human tumor cell lines - Its potential role as an autocrine growth factor SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID SMOOTH-MUSCLE CELLS; GASTRIN-RELEASING PEPTIDE; GENE-ASSOCIATED PEPTIDES; PULMONARY VASCULAR BED; LUNG-CANCER; HYPOTENSIVE PEPTIDE; RAT ADRENOMEDULLIN; COLORIMETRIC ASSAY; NECROSIS-FACTOR; CLONING AB Although adrenomedullin (AM) previously has been identified in human tumors, its role has remained elusive. Analysis by reverse transcriptase-polymerase chain reaction (RT-PCR) revealed AM mRNA in 18 of 20 human normal tissues representing major organs, and 55 of 58 (95%) malignant cell lines. Western blot and high performance liquid chromatography analysis showed immunoreactive AM species of 18, 14, and 6 kDa that are consistent with the precursor, intermediate product, and active peptide, respectively. Immunohistochemistry and in situ RT-PCR performed on paraffin-embedded tumor cell lines of various tissue origins exhibited AM cytoplasmic staining. Neutralizing monoclonal antibody to AM inhibits tumor cell growth in a concentration-dependent manner, an effect that was reversed with the addition of exogenous AM. Responding tumor cells were shown to have approximately 50,000 AM receptors per cell by Scatchard analysis with I-125-AM and expressed AM receptor mRNA by RT-PCR. Our data showed 36 of 48 (75%) tumor cell lines expressed AM receptor mRNA by RT-PCR assessment, all of them also expressed AM. In the presence of AM, cAMP levels were shown to increase in tumor cells. Our collective data demonstrate that AM and AM receptor are expressed in numerous human cancer cell lines of diverse origin and constitute a potential autocrine growth mechanism that could drive neoplastic proliferation. C1 NCI,PEDIAT BRANCH,DIV CANC TREATMENT,NIH,BETHESDA,MD 20892. ARS,USDA,BELTSVILLE,MD 20705. RP Miller, MJ (reprint author), NCI,BIOMARKERS & PREVET RES BRANCH,DIV CLIN SCI,ROCKVILLE,MD 20850, USA. RI Martinez, Alfredo/A-3077-2013 OI Martinez, Alfredo/0000-0003-4882-4044 NR 52 TC 275 Z9 280 U1 1 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD SEP 20 PY 1996 VL 271 IS 38 BP 23345 EP 23351 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VH768 UT WOS:A1996VH76800062 PM 8798536 ER PT J AU Behrens, TW Kearns, GM Rivard, JJ Bernstein, HD Yewdell, JW Staudt, LM AF Behrens, TW Kearns, GM Rivard, JJ Bernstein, HD Yewdell, JW Staudt, LM TI Carboxyl-terminal targeting and novel post-translational processing of JAW1, a lymphoid protein of the endoplasmic reticulum SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID SIGNAL-RECOGNITION PARTICLE; TRANSMEMBRANE PROTEINS; MEMBRANE-PROTEINS; MICROSOMAL-MEMBRANES; MAMMALIAN-CELLS; TRANSLOCATION; SEQUENCE; TRANSPORT; RETENTION; RESIDUES AB Jaw1 is a lymphoid-restricted protein localized to the cytoplasmic face of the endoplasmic reticulum (ER) and is a member of a recently recognized class of integral membrane proteins that contain carboxyl-terminal membrane anchors. The carboxyl-terminal 71 amino acids of the Jaw1 protein, which contain a hydrophobic membrane spanning region, are sufficient to target a heterologous protein to the ER. By discontinuous sucrose gradient ultracentrifugation, differential sedimentation was noted for the four major Jaw1 protein isoforms, with two of the forms predominantly soluble and two microsome-bound. Pulse-chase immunoprecipitations suggest a post translational modification of two major isoforms of the protein resulting in an increase in mobility on SDS-polyacrylamide gel electrophoresis. In. vitro translation studies are compatible with a posttranslational processing event that results in cleavage of a short 36 amino acid lumenal domain. These findings define a carboxyl-terminal domain of the Jaw1 protein that is both necessary and sufficient for ER localization. In addition, the processing of the small lumenal domain of Jaw1 represents a novel post-translational protein modification performed by the endoplasmic reticulum. C1 NCI,METAB BRANCH,BETHESDA,MD 20892. NIDDK,GENET & BIOCHEM BRANCH,BETHESDA,MD 20892. NIH,VIRAL DIS LAB,NIAID,BETHESDA,MD 20892. RP Behrens, TW (reprint author), UNIV MINNESOTA,SCH MED,UMHC,DEPT MED,BOX 108,515 DELAWARE ST SE,MINNEAPOLIS,MN 55455, USA. RI yewdell, jyewdell@nih.gov/A-1702-2012 FU NIAMS NIH HHS [AR01959] NR 37 TC 27 Z9 28 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD SEP 20 PY 1996 VL 271 IS 38 BP 23528 EP 23534 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VH768 UT WOS:A1996VH76800088 PM 8798562 ER PT J AU Witkop, B AF Witkop, B TI Blondes in Venetian paintings, the nine-banded armadillo, and other essays in biochemistry - Bloch,K SO SCIENCE LA English DT Book Review RP Witkop, B (reprint author), NIH,BLDG 10,BETHESDA,MD 20892, USA. NR 1 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 SN 0036-8075 J9 SCIENCE JI Science PD SEP 20 PY 1996 VL 273 IS 5282 BP 1672 EP 1673 PG 2 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VH408 UT WOS:A1996VH40800027 ER PT J AU Drazen, JM Israel, E Boushey, HA Chinchilli, VM Fahy, JV Fish, JE Lazarus, SC Lemanske, RF Martin, RJ Peters, SP Sorkness, C Szefler, SJ AF Drazen, JM Israel, E Boushey, HA Chinchilli, VM Fahy, JV Fish, JE Lazarus, SC Lemanske, RF Martin, RJ Peters, SP Sorkness, C Szefler, SJ TI Comparison of regularly scheduled with as-needed use of albuterol in mild asthma SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Article ID BETA-ADRENERGIC AGONISTS; RANDOMIZED CLINICAL-TRIALS; RESPIRATORY MEDICINE; INHALED SALBUTAMOL; BRONCHIAL-ASTHMA; INTERIM ANALYSES; CONTROVERSIES; THERAPY; HEALTH; MANAGEMENT AB Background Inhaled beta-agonists are the most commonly used treatment for asthma, but data suggest that regularly scheduled use of these agents may have a deleterious effect on the control of asthma. We compared the effects of regularly scheduled use of inhaled albuterol with those of albuterol used only as needed in patients with mild chronic, stable asthma. Methods In a multicenter, double-blind study, we randomly assigned 255 patients with mild asthma to inhale albuterol either on a regular schedule (126 patients) or only as needed (129 patients). The patients were followed for 16 weeks. Results The primary outcome indicator, peak expiratory air flow measured in the morning, did not change significantly during the treatment period in the scheduled (416 liters per minute after the run-in period and 414 liters per minute after the treatment period) or the as-needed (424 liters per minute at both times) treatment groups (P = 0.71). There were no significant differences between the two groups in peak flow variability, forced expiratory volume in one second, the number of puffs of supplemental albuterol needed, asthma symptoms, asthma quality-of-life score, or airway responsiveness to methacholine. The statistically significant differences between the groups in evening peak flow and in the shortterm bronchodilator response to inhaled albuterol were small and judged to be clinically unimportant. Conclusions In patients with mild asthma, neither deleterious nor beneficial effects derived from the regular use of inhaled albuterol beyond those derived from use of the drug as needed. Inhaled albuterol should be prescribed for patients with mild asthma on an as-needed basis. (C) 1996, Massachusetts Medical Society. C1 HARVARD UNIV,SCH MED,BOSTON,MA. UNIV CALIF SAN FRANCISCO,SAN FRANCISCO,CA 94143. MILTON S HERSHEY MED CTR,HERSHEY,PA. THOMAS JEFFERSON UNIV,PHILADELPHIA,PA 19107. UNIV WISCONSIN,MADISON,WI. NATL JEWISH CTR IMMUNOL & RESP MED,DENVER,CO 80206. NHLBI,BETHESDA,MD 20892. RP Drazen, JM (reprint author), BRIGHAM & WOMENS HOSP,RESP DIS DIV,75 FRANCIS ST,BOSTON,MA 02115, USA. FU NHLBI NIH HHS [U10 HL 51834, U10 HL 51831, U10 HL 51843] NR 40 TC 236 Z9 240 U1 0 U2 2 PU MASS MEDICAL SOC PI BOSTON PA 10 SHATTUCK, BOSTON, MA 02115 SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD SEP 19 PY 1996 VL 335 IS 12 BP 841 EP 847 DI 10.1056/NEJM199609193351202 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA VG869 UT WOS:A1996VG86900002 PM 8778601 ER PT J AU Das, R Vonderhaar, BK AF Das, R Vonderhaar, BK TI Involvement of SHC, GRB2, SOS and RAS in prolactin signal transduction in mammary epithelial cells SO ONCOGENE LA English DT Article DE prolactin; breast cancer; JAK2; SHC; STAT5 ID HUMAN-BREAST-CANCER; ACTIVATED PROTEIN-KINASE; GROWTH-HORMONE RECEPTOR; TYROSINE PHOSPHORYLATION; TRANSCRIPTION FACTOR; GENE-EXPRESSION; LYMPHOMA-CELLS; MAP KINASE; ASSOCIATION; JAK2 AB The peptide hormone prolactin (Prl) regulates proliferation of normal and malignant mammary cells, In the present study me demonstrate that tate Prl responsive cell Lines, NOG-8 and T47D, activate the JAK2-SHC-MAPK pathway in a rapid and transient manner, Within 1 min of Prl treatment there was an increase in association of JAK2 with SHC, followed by rapid phosphorylation of both the 52 kDa and 46 kDa SHC proteins, Grb2 and Sos associated with the SHC proteins within 1-3 min of Prl treatment in these mammary cells, Within 5 min of hormone treatment we observe an increase in ras-GTP suggesting activation of ras, We also showed a rapid and transient tyrosine phosphorylation of STAT5 in proliferating T47D cells which reached its peak after 30 min of Prl treatment, These results indicate that Prl receptors, after binding the ligand, activate several pathways for signal transduction leading to mitogenesis. C1 NCI,TUMOR IMMUNOL & BIOL LAB,NIH,BETHESDA,MD 20892. NR 40 TC 73 Z9 73 U1 0 U2 0 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE, HAMPSHIRE, ENGLAND RG21 6XS SN 0950-9232 J9 ONCOGENE JI Oncogene PD SEP 19 PY 1996 VL 13 IS 6 BP 1139 EP 1145 PG 7 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA VJ202 UT WOS:A1996VJ20200004 PM 8808687 ER PT J AU Gale, NW Flenniken, A Compton, DC Jenkins, N Copeland, NG Gilbert, DJ Davis, S Wilkinson, DG Yancopoulos, GD AF Gale, NW Flenniken, A Compton, DC Jenkins, N Copeland, NG Gilbert, DJ Davis, S Wilkinson, DG Yancopoulos, GD TI Elk-L3, a novel transmembrane ligand for the Eph family of receptor tyrosine kinases, expressed in embryonic floor plate, roof plate and hindbrain segments SO ONCOGENE LA English DT Article DE Eph; LERK; Sek; Elk; Nuk; Htk; floor plate; roof plate ID INSITU HYBRIDIZATION; SONIC-HEDGEHOG; MIDLINE CELLS; MOUSE; GENE; ORGANIZATION; MEMBER; MAP; MOLECULES; POLARITY AB The Eph family of receptor tyrosine kinases has 13 distinct members and seven ligands for these receptors have been described to date, These receptors and their ligands have been implicated in regulating neuronal axon guidance and in patterning of the developing nervous system and may also serve a patterning and compartmentalization role outside of the nervous system as well, The ligands are all membrane-attached, and this attachment appears to be crucial for their normal function; five of the known ligands are linked to the membrane,ia a glycosyl phosphotidylinositol (GPI) linkage, while tate of the ligands are transmembrane proteins, Despite the large number of Eph family receptors and ligands, they can be divided into just two major subclasses based on their binding specificities. All the GPI-anchored ligands bind and activate one subclass of the Eph receptors (that represented by Eck) while the two transmembrane ligands bind and activate the other major subclass of receptors (represented by Elk), Here we report the identification and characterization of the third, and most divergent, member of the transmembrane group of Eph ligands, which we term Elk-L3 (ElK-related receptor ligand number 3), Elk-L3 is notable for its remarkably restricted and prominent expression in the Boor plate and roof plate of the developing neural tube and its rhombomere-specific expression in the developing hindbrain, The Elk-L3 gene has been localized to mouse chromosome 11 and human chromosome 17. C1 REGENERON PHARMACEUT INC,TARRYTOWN,NY 10591. NATL INST MED RES,MRC,DIV DEV NEUROBIOL,LONDON NW7 1AA,ENGLAND. NCI,FREDERICK CANC RES & DEV CTR,ABL,MAMMALIAN GENET LAB,FREDERICK,MD 21702. NR 39 TC 103 Z9 104 U1 0 U2 2 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE, HAMPSHIRE, ENGLAND RG21 6XS SN 0950-9232 J9 ONCOGENE JI Oncogene PD SEP 19 PY 1996 VL 13 IS 6 BP 1343 EP 1352 PG 10 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA VJ202 UT WOS:A1996VJ20200026 PM 8808709 ER PT J AU Fried, LP Francomano, CA MacDonald, SM Wagner, EM Stokes, EJ Carbone, KM Bias, WB Newman, MM Stobo, JD AF Fried, LP Francomano, CA MacDonald, SM Wagner, EM Stokes, EJ Carbone, KM Bias, WB Newman, MM Stobo, JD TI Career development for women in academic medicine - Multiple interventions in a department of medicine SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID PHYSICIANS; PROFESSION; PROMOTION; SCIENCE; FACULTY; MEN AB Objective.-To determine the gender-based career obstacles for women in an academic department of medicine and to report the interventions to correct such obstacles (resulting from the evaluation) and the results of these interventions, Design.-Intervention study, before-after trial, with assessment of faculty concerns and perceived change through structured, self-administered questionnaires. Setting.-The Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Md. Participants.-Full-time faculty. Interventions.-Multifaceted intervention from 1990 through 1995 to correct gender-based career obstacles reported by women faculty, including problem identification, leadership, and education of faculty, and interventions to improve faculty development, mentoring, and rewards and to reduce isolation and structural career impediments. Main Outcome Measures.-Retention and promotion of deserving women faculty, salary equity, quality of mentoring, decreased isolation from information and colleagues, integration of women faculty into the scientific community, and decreased manifestations of gender bias. Results.-Junior women were retained and promoted, reversing previous experience, with a 550% increase in the number of women at the associate professor rank over 5 years (from 4 in 1990 to 26 in 1995), Interim 3-year follow-up showed a 183% increase in the proportion of women faculty who expected they would still be in academic medicine in 10 years (from 23% [7/30] in 1990 to 65% [30/46] in 1993). One half to two thirds of women faculty reported improvements in timeliness of promotions, manifestations of gender bias, access to information needed for faculty development, isolation, and salary equity, Men also reported improvements in these areas. Conclusions.-The outcomes reported here indicate that it is possible to make substantive improvements in the development of women's careers, that an institutional strategy to this end can be successful in retaining women in academic medicine, and that such interventions are likely to benefit all faculty. Long-term interventions appear essential. C1 JOHNS HOPKINS UNIV,SCH MED,DEPT MED,BALTIMORE,MD 21205. NIH,NATL CTR HUMAN GENOME RES,MED GENET BRANCH,BETHESDA,MD 20892. JOHNS HOPKINS HLTHCARE LLC,BALTIMORE,MD. NR 41 TC 152 Z9 152 U1 1 U2 17 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD SEP 18 PY 1996 VL 276 IS 11 BP 898 EP 905 DI 10.1001/jama.276.11.898 PG 8 WC Medicine, General & Internal SC General & Internal Medicine GA VF787 UT WOS:A1996VF78700033 PM 8782639 ER PT J AU Kohn, KW AF Kohn, KW TI Regulatory genes and drug sensitivity SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Editorial Material ID APOPTOSIS; RESISTANCE RP Kohn, KW (reprint author), NCI,NIH,MOL PHARMACOL LAB,DIV BASIC SCI,BLGD 37,RM 5C25,BETHESDA,MD 20892, USA. NR 15 TC 11 Z9 13 U1 0 U2 0 PU NATL CANCER INSTITUTE PI BETHESDA PA 9030 OLD GEORGETOWN RD, BETHESDA, MD 20814 SN 0027-8874 J9 J NATL CANCER I JI J. Natl. Cancer Inst. PD SEP 18 PY 1996 VL 88 IS 18 BP 1255 EP 1256 DI 10.1093/jnci/88.18.1255 PG 2 WC Oncology SC Oncology GA VG673 UT WOS:A1996VG67300001 PM 8797761 ER PT J AU Torrey, EF Rawlings, RR Ennis, JM Merrill, DD Flores, DS AF Torrey, EF Rawlings, RR Ennis, JM Merrill, DD Flores, DS TI Birth seasonality in bipolar disorder, schizophrenia, schizoaffective disorder and stillbirths SO SCHIZOPHRENIA RESEARCH LA English DT Article DE seasonality; time series; schizophrenia; bipolar; schizoaffective; depression; stillbirths ID MENTAL DISORDER; GENERAL POPULATION; AGE INCIDENCE; PSYCHOSES; SUBTYPES AB Background: More than 40 studies have been done on seasonal birth patterns for schizophrenia, but only two small studies have been done for DSM-III-R bipolar disorder and none for schizoaffective disorder. Two studies have also reported a significant relationship between schizophrenia births and stillbirths. Methods. In the largest study to date, birth data from four states was obtained on 126 987 state psychiatric hospitals inpatients divided into 'process' schizophrenia (disorganized, catatonic, undifferentiated), paranoid schizophrenia, schizoaffective disorder, bipolar disorder and major depression. Time series analysis compared these births to all general births and to stillbirths. Results: 'Process' schizophrenia, paranoid schizophrenia, schizoaffective disorder and bipolar disorder all had statistically significant seasonal excess births from December through March (p=0.0000). The largest excess was 5.8% for bipolar disorder. Major depression had significant excess births from March through May. Time series analysis showed statistically significant coherences between major depression and bipolar disorder (0.995) and between schizoaffective disorder and both 'process' schizophrenia (0.977) and bipolar disorder (0.977). Unexpectedly, a significant coherence was also found between paranoid schizophrenia and bipolar disorder (0.972). Excess stillbirths were found for each month from January through June and a significant coherence was found between stillbirths and paranoid schizophrenia (0.998). Conclusions: This study demonstrates that DSM-III-R bipolar disorder and schizoaffective disorder both have an excess of winter births, similar to that found in schizophrenia. Time series analysis, however, suggests that the causes may not be identical. Major depression, by contrast, has an excess of spring births. C1 NIAAA,CLIN STUDIES LAB,BETHESDA,MD 20892. VIRGINIA DEPT MENTAL HLTH,MENTAL RETARDAT & SUBSTANCE ABUSE SERV,RICHMOND,VA 23219. N CAROLINA DEPT HUMAN RESOURCES,DEV DISABIL & SUBSTANCE ABUSE SERV,DIV MENTAL HLTH,RALEIGH,NC 27603. PENN DEPT PUBL WELF,OFF MENTAL HLTH,DIV PLANNING EVALUAT & SYST,HARRISBURG,PA 17105. RP Torrey, EF (reprint author), ST ELIZABETH HOSP,NIMH,CTR NEUROSCI,2700 MARTIN LUTHER KING JR AVE SE,WASHINGTON,DC 20032, USA. NR 39 TC 67 Z9 68 U1 2 U2 11 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0920-9964 J9 SCHIZOPHR RES JI Schizophr. Res. PD SEP 18 PY 1996 VL 21 IS 3 BP 141 EP 149 PG 9 WC Psychiatry SC Psychiatry GA VJ171 UT WOS:A1996VJ17100002 PM 8885042 ER PT J AU Kang, MS Lee, HJ Lee, JH Ku, JL Lee, KP Kelley, MJ Won, YJ Kim, ST Park, JG AF Kang, MS Lee, HJ Lee, JH Ku, JL Lee, KP Kelley, MJ Won, YJ Kim, ST Park, JG TI Mutation of p53 gene in hepatocellular carcinoma cell lines with HBX DNA SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article ID HUMAN LUNG-CANCER; VIRUS-X PROTEIN; LIVER-CANCER; POLYMORPHISM; EXPRESSION; ANTIGEN; HETEROGENEITY; P53-GENE; MICE AB It has been known that the incidence of p53 mutation is very rare in HEX-positive primary human hepatocellular carcinoma (HCC) tissues. The frequency of p53 mutation, however, in established cell lines with integrated HEV DNA and/or HEX has not been well studied. To know p53 mutational frequency, and to investigate whether the presence of HEX DNA sequence correlates with the absence of p53 mutation in the established HCC cell lines, we studied the p53 mutation and the presence of HEX sequence in IZ recently characterized HCC cell lines. As a result, all 12 (100%) lines showed mutation in the p53 gene. Three (25%) cell lines had transversion of codon 215 while no mutation of codon 249 was found. In contrast with previous reports, although HEX DNA was present in 11 cell lines, p53 mutation had occurred, indicating that the presence of HEX viral DNA does not correlate with a lack of p53 mutation in established HCC cell lines. Our results suggest that the frequency of p53 mutation is extremely high even in HEX DNA positive HCC cell lines. C1 SEOUL NATL UNIV,COLL MED,CELL BIOL LAB,CANC RES INST,SEOUL 110799,SOUTH KOREA. SEOUL NATL UNIV,COLL MED,CELL BIOL LAB,CANC RES CTR,SEOUL 110799,SOUTH KOREA. NCI,NAVY MED ONCOL BRANCH,NATL CANC RES,BETHESDA,MD 20892. RI Ku, Ja-Lok/D-5736-2012; Park, Jae-Gahb/J-5494-2012; OI Kelley, Michael/0000-0001-9523-6080 NR 36 TC 32 Z9 33 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC 605 THIRD AVE, NEW YORK, NY 10158-0012 SN 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD SEP 17 PY 1996 VL 67 IS 6 BP 898 EP 902 DI 10.1002/(SICI)1097-0215(19960917)67:6<898::AID-IJC22>3.0.CO;2-X PG 5 WC Oncology SC Oncology GA VK490 UT WOS:A1996VK49000022 PM 8824565 ER PT J AU Boisclair, YR Seto, D Hsieh, S Hurst, KR Ooi, GT AF Boisclair, YR Seto, D Hsieh, S Hurst, KR Ooi, GT TI Organization and chromosomal localization of the gene encoding the mouse acid labile subunit of the insulin-like growth factor binding complex SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE 150-kDa insulin-like growth factor complex; insulin-like growth factor binding protein; growth hormone ID PROMOTED TYROSYL PHOSPHORYLATION; PROTEIN COMPLEX; FACTOR-I; HORMONE; RAT; TRANSCRIPTION; EXPRESSION; SERUM; PURIFICATION; INITIATION AB After birth, most of insulin-like growth factor I and II (IGFs) circulate as a ternary complex formed by the association of IGF binding protein 3-IGF complexes with a serum protein called acid-labile subunit (ALS). ALS retains the IGF binding protein-3-IGF complexes in the vascular compartment and extends the t(1/2) of IGFs in the circulation. Synthesis of ALS occurs mainly in liver after birth and is stimulated by growth hormone. To study the basis for this regulation, we cloned and characterized the mouse ALS gene, Comparison of genomic and cDNA sequences indicated that the gene is composed of two exons separated by a 1126-bp intron. Exon 1 encodes the first 5 amino acids of the signal peptide and contributes the first nucleotide of codon 6. Exon 2 contributes the last 2 nt of codon 6 and encodes the remaining 17 amino acids of the signal peptide as well as the 580 amino acids of the mature protein. The polyadenylylation signal, ATTAAA, is located 241 bp from the termination codon. The cDNA and genomic DNA diverge 16 bp downstream from this signal. Transcription initiation was mapped to 11 sites over a 140-bp TATA-less region. The DNA fragment extending from nt -805 to -11 (ATG, +1) directed basal and growth hormone-regulated expression of a luciferase reporter plasmid in the rat liver cell line H4-II-E. Finally, the ALS gene was mapped to mouse chromosome 17 by fluorescence in situ hybridization. C1 DUPONT MERCK PHARMACEUT CO,WILMINGTON,DE 19880. NIH,GROWTH & DEV SECT,MOL & CELL ENDOCRINOL BRANCH,NIDDKD,BETHESDA,MD 20892. RP Boisclair, YR (reprint author), CORNELL UNIV,DEPT ANIM SCI,ITHACA,NY 14853, USA. NR 37 TC 31 Z9 33 U1 0 U2 0 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD SEP 17 PY 1996 VL 93 IS 19 BP 10028 EP 10033 DI 10.1073/pnas.93.19.10028 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VJ203 UT WOS:A1996VJ20300009 PM 8816745 ER PT J AU Silva, AM Lee, AY Gulnik, SV Majer, P Collins, J Bhat, TN Collins, PJ Cachau, RE Luker, KE Gluzman, IY Francis, SE Oksman, A Goldberg, DE Erickson, JW AF Silva, AM Lee, AY Gulnik, SV Majer, P Collins, J Bhat, TN Collins, PJ Cachau, RE Luker, KE Gluzman, IY Francis, SE Oksman, A Goldberg, DE Erickson, JW TI Structure and inhibition of plasmepsin II, a hemoglobin-degrading enzyme from Plasmodium falciparum SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE malaria; drug design; crystallography; aspartic protease; cathepsin D ID ASPARTIC PROTEINASE; EXPRESSION; MALARIA; TARGET; PEPSIN AB Plasmodium falciparum is the major causative agent of malaria, a disease of worldwide importance, Resistance to current drugs such as chloroquine and mefloquine is spreading at an alarming rate, and our antimalarial armamentarium is almost depleted, The malarial parasite encodes two homologous aspartic proteases, plasmepsins I and II, which are essential components of its hemoglobin-degradation pathway and are novel targets for antimalarial drug development, We have determined the crystal structure of recombinant plasmepsin II complexed with pepstatin A. This represents the first reported crystal structure of a protein from P. falciparum. The crystals contain molecules in two different conformations, revealing a remarkable degree of interdomain flexibility of the enzyme. The structure was used to design a series of selective low molecular weight compounds that inhibit both plasmepsin II and the growth of P. falciparum in culture. C1 NCI, STRUCT BIOCHEM PROGRAM, SAIC, FREDERICK, MD 21702 USA. WASHINGTON UNIV, SCH MED, HOWARD HUGHES MED INST, DEPT MOL MICROBIOL, ST LOUIS, MO 63110 USA. WASHINGTON UNIV, SCH MED, HOWARD HUGHES MED INST, DEPT MED, ST LOUIS, MO 63110 USA. WASHINGTON UNIV, SCH MED, JEWISH HOSP ST LOUIS, ST LOUIS, MO 63110 USA. FU NCI NIH HHS [N01 CO-56000]; NIAID NIH HHS [AI 37977] NR 28 TC 211 Z9 216 U1 0 U2 9 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 SEP 17 PY 1996 VL 93 IS 19 BP 10034 EP 10039 DI 10.1073/pnas.93.19.10034 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VJ203 UT WOS:A1996VJ20300010 PM 8816746 ER PT J AU Hunyady, L Zhang, M Jagadeesh, G Bor, M Balla, T Catt, KJ AF Hunyady, L Zhang, M Jagadeesh, G Bor, M Balla, T Catt, KJ TI Dependence of agonist activation on a conserved apolar residue in the third intracellular loop of the AT(1) angiotensin receptor SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE G protein-coupled receptors; inositol phosphate signaling; receptor internalization; site-directed mutagenesis ID BETA-ADRENERGIC-RECEPTOR; ADRENAL GLOMERULOSA CELLS; PROTEIN-COUPLED RECEPTORS; SMOOTH-MUSCLE CELLS; II RECEPTOR; INDUCED INTERNALIZATION; SIGNAL-TRANSDUCTION; ALDOSTERONE SECRETION; POTENTIAL MECHANISM; TYROSINE RESIDUE AB The coupling of agonist-activated seven transmembrane domain receptors to G proteins is known to involve the amino-terminal region of their third cytoplasmic loop. Analysis of the amino acids in this region of the rat type 1a angiotensin (AT(1a)) receptor identified Leu-222 as an essential residue in receptor activation by the physiological agonist, angiotensin II (Ang II). Nonpolar replacements for Leu-222 yielded functionally intact AT(1) receptors, while polar or charged residues caused progressive impairment of Ang II-induced inositol phosphate generation. The decrease in agonist-induced signal generation was associated with a parallel reduction of receptor internalization, and was most pronounced for the Lys-222 mutant receptor. Although this mutant showed normal binding of the peptide antagonist, [Sar(1),Ile(8)]Ang II, its affinity for Ang II was markedly reduced, consistent with its inability to adopt the high-affinity conformation. A search revealed that many G(q)-coupled receptors contain an apolar amino acid (frequently leucine) in the position corresponding to Leu-222 of the AT(1) receptor. These findings suggest that such a conserved apolar residue in the third intracellular loop is a crucial element in the agonist-induced activation of the AT(1) and possibly many other G protein-coupled receptors. C1 NIH,ENDOCRINOL & REPROD RES BRANCH,NICHHD,BETHESDA,MD 20892. US FDA,DIV CARDIO RENAL DRUG PROD,CTR DRUG EVALUAT & RES,ROCKVILLE,MD 20857. OI Balla, Tamas/0000-0002-9077-3335 NR 54 TC 33 Z9 34 U1 1 U2 1 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD SEP 17 PY 1996 VL 93 IS 19 BP 10040 EP 10045 DI 10.1073/pnas.93.19.10040 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VJ203 UT WOS:A1996VJ20300011 PM 8816747 ER PT J AU Mushegian, AR Koonin, EV AF Mushegian, AR Koonin, EV TI A minimal gene set for cellular life derived by comparison of complete bacterial genomes SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID ESCHERICHIA-COLI; SEQUENCE DATABASES; PROTEINS; COMMON AB The recently sequenced genome of the parasitic bacterium Mycoplasma genitalium contains only 468 identified protein-coding genes that have been dubbed a minimal gene complement [Fraser, C. M., Gocayne, J. D., White, O., Adams, M. D., Clayton, R. A., et al. (1995) Science 270, 397-403]. Although the M. genitalium gene complement is indeed the smallest among known cellular life forms, there is no evidence that it is the minimal self-sufficient gene set, To derive such a set, we compared the 468 predicted M. genitalium protein sequences with the 1703 protein sequences encoded by the other completely sequenced small bacterial genome, that of Haemophilus influenzae. M. genitalium and H. influenzae belong to two ancient bacterial lineages, i.e., Gram-positive and Gram-negative bacteria, respectively. Therefore, the genes that are conserved in these two bacteria are almost certainly essential for cellular function. It is this category of genes that is most likely to approximate the minimal gene set. We found that 240 M. genitalium genes have orthologs among the genes of H. influenzae. This collection of genes falls short of comprising the minimal set as some enzymes responsible for intermediate steps in essential pathways are missing. The apparent reason for this is the phenomenon that we call nonorthologous gene displacement when the same function is fulfilled by nonorthologous proteins in two organisms. We identified 22 nonorthologous displacements and supplemented the set of orthologs with the respective M. genitalium genes. After examining the resulting List of 262 genes for possible functional redundancy and for the presence of apparently parasite-specific genes, 6 genes were removed. We suggest that the remaining 256 genes are close to the minimal gene set that is necessary and sufficient to sustain the existence of a modern-type cell. Most of the proteins encoded by the genes from the minimal set have eukaryotic or archaeal homologs but seven key proteins of DNA replication do not. We speculate that the last common ancestor of the three primary kingdoms had an RNA genome. Possibilities are explored to further reduce the minimal set to model a primitive cell that might have existed at a very early stage of life evolution. C1 NATL LIB MED,NATL CTR BIOTECHNOL INFORMAT,NIH,BETHESDA,MD 20894. OI Mushegian, Arcady/0000-0002-6809-9225 NR 34 TC 539 Z9 568 U1 4 U2 39 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD SEP 17 PY 1996 VL 93 IS 19 BP 10268 EP 10273 DI 10.1073/pnas.93.19.10268 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VJ203 UT WOS:A1996VJ20300052 PM 8816789 ER PT J AU Frank, EG Ennis, DG Gonzalez, M Levine, AS Woodgate, R AF Frank, EG Ennis, DG Gonzalez, M Levine, AS Woodgate, R TI Regulation of SOS mutagenesis by proteolysis SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE UmuD; D'; UmuC; Lon; ClpXP; protein-protein interactions ID CAPSULAR POLYSACCHARIDE SYNTHESIS; DEPENDENT CLP PROTEASE; ESCHERICHIA-COLI; UV MUTAGENESIS; SPECIFICITY COMPONENT; MOLECULAR CHAPERONE; BACTERIOPHAGE-MU; SERINE-PROTEASE; UMUD CLEAVAGE; GROE MUTANTS AB DNA damage-inducible mutagenesis in Escherichia coli is largely dependent upon the activity of the UmuD (UmuD') and UmuC proteins. The intracellular level of these proteins is tightly regulated at both the transcriptional and the posttranslational levels. Such regulation presumably allows cells to deal with DNA damage via error-free repair pathways before being committed to error-prone pathways. We have recently discovered that as part of this elaborate regulation, both the UmuD and the UmuC proteins are rapidly degraded in vivo. We report here that the enzyme responsible for their degradation is the ATP-dependent serine protease, Lon. In contrast, UmuD' (the posttranslational product and mutagenically active form of UmuD) is degraded at a much reduced rate by Lon, but is instead rapidly degraded by another ATP-dependent protease, ClpXP. Interestingly, UmuD' is rapidly degraded by ClpXP only when it is in a heterodimeric complex with UmuD. Formation of UmnD/UmuD' heterodimers in preference to UmuD' homodimers therefore targets UmuD' protein for proteolysis. Such a mechanism allows cells to reduce the intracellular levels of the mutagenically active Umu proteins and thereby return to a resting state once error-prone DNA repair has occurred. The apparent half-life of the heterodimeric UmuD/D' complex is greatly increased in the clpX::Kan and clpP::Kan strains and these strains are correspondingly rendered virtually UV non-mutable. We believe that these phenotypes are consistent with the suggestion that while the UmuD/D' heterodimer is mutagenically inactive, it still retains the ability to interact with UmuC, and thereby precludes the formation of the mutagenically active UmuD'C-2 complex. C1 NICHHD,SECT DNA REPLICAT REPAIR & MUTAGENESIS,NIH,BETHESDA,MD 20892. UNIV SW LOUISIANA,DEPT BIOL,LAFAYETTE,LA 70504. CALTECH,DIV BIOL,PASADENA,CA 91125. NR 47 TC 121 Z9 125 U1 0 U2 0 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD SEP 17 PY 1996 VL 93 IS 19 BP 10291 EP 10296 DI 10.1073/pnas.93.19.10291 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VJ203 UT WOS:A1996VJ20300056 PM 8816793 ER PT J AU Racioppi, L Matarese, G DOro, U DePascale, M Masci, AM Fontana, S Zappacosta, S AF Racioppi, L Matarese, G DOro, U DePascale, M Masci, AM Fontana, S Zappacosta, S TI The role of CD4-Lck in T-cell receptor antagonism: Evidence for negative signaling SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID CD45 PHOSPHOTYROSINE PHOSPHATASE; TYROSINE PROTEIN-KINASE; ANTIGEN RECOGNITION; THYMIC SELECTION; CROSS-LINKING; ACTIVATION; LIGAND; PHOSPHORYLATION; TRANSDUCTION; ASSOCIATION AB Small changes in the complex between a peptide and a molecule of the major histocompatibilty complex generate ligands able to partially activate (partial agonist) or even inhibit (antagonist) T-cell functions. T-cell receptor engagement of antagonist complex results in a partial zeta chain phosphorylation without activation of the associated ZAP-70 kinase. Herein we show that, despite a strong inhibition of both inositol phospholipid hydrolysis and extracellular regulated kinase activation, exposure of a T-cell clone to increasing antagonist concentrations increased the activity of the CD4-Lck kinase. Addition of anti-CD4 antibody to culture medium prevented inhibitory effects induced by antagonist ligand, We propose that CD4-Lck activation triggered by antagonist complexes may act in a dominant negative mode, thus overriding stimulatory signals coming from agonist ligand. These findings identify a new T-cell signaling profile that may explain the ability of some T-cell receptor variant ligands to inhibit specific biological activities or trigger alternative activation programs. C1 NCI,LAB IMMUNE CELL BIOL,BETHESDA,MD 20892. CNR,CTR ENDOCRINOL & ONCOL SPERIMENTALE,I-80131 NAPLES,ITALY. RP Racioppi, L (reprint author), UNIV NAPLES,DIPARTIMENTO BIOL & PATOL CELLULARE & MOL,CATTEDRA IMMUNOL,5 VIA S PANSINI,I-80131 NAPLES,ITALY. OI racioppi, luigi/0000-0002-9207-6752; Masci, Anna Maria/0000-0003-1940-6740; Matarese, Giuseppe/0000-0001-9429-0616 NR 43 TC 26 Z9 26 U1 0 U2 0 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD SEP 17 PY 1996 VL 93 IS 19 BP 10360 EP 10365 DI 10.1073/pnas.93.19.10360 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VJ203 UT WOS:A1996VJ20300068 PM 8816805 ER PT J AU Mezey, E Eisenhofer, G Harta, G Hansson, S Gould, L Hunyady, B Hoffman, BJ AF Mezey, E Eisenhofer, G Harta, G Hansson, S Gould, L Hunyady, B Hoffman, BJ TI A novel nonneuronal catecholaminergic system: Exocrine pancreas synthesizes and releases dopamine SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE monoamine transporters; mucosal healing; dopamine receptor ID TYROSINE-HYDROXYLASE; MESSENGER-RNA; RAT; EXPRESSION; SECRETION; TRANSPORTER; AGONISTS; RECEPTOR; CLONING; RESPONSES AB Cells of the exocrine pancreas produce digestive enzymes potentially harmful to the intestinal mucosa. Dopamine has been reported to protect against mucosal injury, In looking for the source of dopamine in the small intestine, we found that the duodenal juice contains high levels of dopamine and that the pancreas itself has a high dopamine [and dihydroxyphenylalanine (dopa)] content that does not change significantly after chemical sympathectomy. Furthermore, we were able to demonstrate tyrosine hydroxylase (TH) activity in control pancreas as well as in pancreas from rats after chemical sympathectomy. Immunostaining and in situ hybridization histochemistry confirmed both the presence of TH, dopamine, and the dopamine transporter, and the mRNAs encoding TH and dopamine transporter, and the presence of both types of vesicular monoamine transporters in the exocrine cells of the pancreas. Since there are no catecholaminergic enteric ganglia in the pancreas, the above results indicate that pancreatic cells have all the characteristics of dopamine-producing cells. We suggest that the pancreas is an important source of nonneuronal dopamine in the body, and that this dopamine has a role in protecting the intestinal mucosa and suggests that dopamine D1b receptor agonists might be used to help mucosal healing in the gastrointestinal tract. C1 NIMH,CELL BIOL LAB,BETHESDA,MD 20892. RP Mezey, E (reprint author), NINCDS,CLIN NEUROSCI BRANCH,BLDG 36,3A17,BETHESDA,MD 20892, USA. NR 39 TC 67 Z9 67 U1 0 U2 2 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD SEP 17 PY 1996 VL 93 IS 19 BP 10377 EP 10382 DI 10.1073/pnas.93.19.10377 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VJ203 UT WOS:A1996VJ20300071 PM 8816808 ER PT J AU Wang, JY Mushegian, A Lory, S Jin, SG AF Wang, JY Mushegian, A Lory, S Jin, SG TI Large-scale isolation of candidate virulence genes of Pseudomonas aeruginosa by in vivo selection SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE in vivo expression; IVET; neutropenic mouse; purEK; Fur ID BORDETELLA-PERTUSSIS; NUCLEOTIDE-SEQUENCE; ESCHERICHIA-COLI; REPRESSED GENE; LIVE-VACCINE; CLONING; IRON; FUR; SIDEROPHORE; DETERMINANT AB Pseudomonas aeruginosa, an opportunistic human pathogen, is a major causative agent of mortality and morbidity in immunocompromised patients and those with cystic fibrosis genetic disease. To identify new virulence genes of P. aeruginosa, a selection system was developed based on the in pipe expression technology (IVET) that was first reported in Salmonella system. An adenine-requiring auxotrophic mutant strain of P. aeruginosa was isolated and found avirulent on neutropenic mice. A DNA fragment that ran complement the mutant strain, containing purEK operon that is required for de novo biosynthesis of purine, was sequenced and used in the IVET vector construction. By applying the IVET selection system to a neutropenic mouse infection model, genetic loci that are specifically induced in vivo were identified. Twenty-two such loci were partially sequenced and analyzed. One of them was a well-studied virulence factor, pyochelin receptor (FptA), that is involved in iron acquisition Fifteen showed significant homology to reported sequences in GenBank, while the remaining six did not. One locus, designated np20, encodes an open reading frame that shares amino acid sequence homology to transcriptional regulators, especially to the ferric uptake regulator (Fur) proteins of other bacteria. An insertional np20 null mutant strain of P. aeruginosa did not show a growth defect on laboratory media; however, its virulence on neutropenic mice was significantly reduced compared with that of a wild-type parent strain, demonstrating the importance of the np20 locus in the bacterial virulence. The successful isolation of genetic loci that affect bacterial virulence demonstrates the utility of the IVET system in identification of new virulence genes of P. aeruginosa. C1 UNIV ARKANSAS MED SCI HOSP,DEPT MICROBIOL & IMMUNOL,LITTLE ROCK,AR 72205. NATL LIB MED,NATL CTR BIOTECHNOL INFORMAT,NIH,BETHESDA,MD 20894. UNIV WASHINGTON,DEPT MICROBIOL,SEATTLE,WA 98195. OI Mushegian, Arcady/0000-0002-6809-9225 NR 37 TC 128 Z9 143 U1 1 U2 2 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD SEP 17 PY 1996 VL 93 IS 19 BP 10434 EP 10439 DI 10.1073/pnas.93.19.10434 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VJ203 UT WOS:A1996VJ20300081 PM 8816818 ER PT J AU Lin, MQ Nash, HA AF Lin, MQ Nash, HA TI Influence of general anesthetics on a specific neural pathway in Drosophila melanogaster SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE electrophysiology; synaptic transmission; halothane ID GIANT NERVE-FIBER; CAENORHABDITIS-ELEGANS; VOLATILE ANESTHETICS; MUTANTS; FLIGHT; SENSITIVITY; NEURONS AB The neural pathway that governs an escape response of Drosophila to sudden changes in light intensity can be artificially induced by electrical stimulation of the brain and monitored by electrical recording from the effector muscles. We have refined previous work in this system to permit reliable ascertainment of two kinds of response: (i) a short-latency response that follows from direct excitation of a giant fiber neuron in the interior of the fly brain and (ii) a long-latency response in which electrical stimulation triggers neurons in the optic ganglia that ultimately impinge on the giant fiber neuron. The general anesthetic halothane is reported here to have very different potencies in inhibiting these two responses. The long-latency response is obliterated at concentrations similar to those that cause gross behavioral effects in adult flies, whereas the short-latency response is only partially inhibited at doses that are 10-fold higher. Three other volatile anesthetic agents show a similar pattern. Thus, as in higher organisms, the Drosophila nervous system is differentiated into components of high and low sensitivity to general anesthetics. Moreover, this work shows that one of the sensitive components of the nervous system lies in the optic lobe and is readily assayed by its effect on downstream systems; it should provide a focus for exploring the effects of genetic alteration of anesthetic sensitivity. RP Lin, MQ (reprint author), NIMH,MOL BIOL LAB,BETHESDA,MD 20892, USA. NR 30 TC 24 Z9 24 U1 1 U2 1 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD SEP 17 PY 1996 VL 93 IS 19 BP 10446 EP 10451 DI 10.1073/pnas.93.19.10446 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA VJ203 UT WOS:A1996VJ20300083 PM 8816820 ER PT J AU Nishida, K Markey, SP AF Nishida, K Markey, SP TI Nicardipine and MK-801 attenuate platelet-activating factor increases following cerebral ischemia-reperfusion in gerbils SO BRAIN RESEARCH LA English DT Article DE platelet-activating factor; brain ischemia; nicardipine; bay K8644; MK-801; Ca2+ influx ID D-ASPARTATE ANTAGONIST; RAT-BRAIN; CALCIUM CHANNELS; NEURONAL DAMAGE; FOREBRAIN ISCHEMIA; ARTERY OCCLUSION; HIPPOCAMPUS; INJURY; NILVADIPINE; CA-2+ AB The effects of pretreatment with nicardipine (dihydropyridine Ca2+ channel antagonist), Bay K8634 (dihydropyridine Ca2+ channel agonist), and MK-801 (N-methyl-D-aspartate-receptor antagonist) on changes of platelet-activating factor (PAF) concentrations in transient ischemic brain are reported. The tissue concentration of PAF increases significantly in hippocampus, cortex and thalamus by 210%, 169% and 168% of controls without ischemia-reperfusion, respectively after 1 h of reperfusion. Nicardipine (5 mg/kg) reduces the accumulation of PAF, the remaining increases in hippocampus, cortex and thalamus being 151%, 138% and 145% of the controls, respectively. in contrast, Bay K8644 (2.5 mg/kg) enhances the accumulation of PAF, its concentrations in hippocampus, cortex and thalamus being 376%, 233% and 204% of the controls, respectively. The Bay K8644 enhancement in hippocampus is completely inhibited by pretreatment of nicardipine (5 mg/kg). MK-801 (10 mg/kg) reduces the accumulation of PAF, the remaining increases in hippocampus, cortex and thalamus being 152%, 147% and 144% of the controls, respectively. Moreover, brain tissue from animals subjected to the combined pretreatment with nicardipine (5 mg/kg) and MK-801 (10 mg/kg) indicates there is greater inhibition of ischemia-induced PAF increases than with either drug alone. These results indicate that PAF production in the ischemic brain may be regulated by Ca2+ influx through voltage-sensitive Ca2+ channels which are antagonized and agonized by nicardipine and Bay K8644, respectively and receptor-operated Ca2+ channels which are antagonized by MK-801. Because it is known that increases of intracellular Ca2+ in the brain accompany ischemia and early periods of reperfusion and that PA-F exhibits neurotoxicity, the present findings support the role of PAF as a mediator in ischemia-induced brain damage at early stages of reperfusion. C1 NIMH, ANALYT BIOCHEM SECT, CLIN SCI LAB, BETHESDA, MD 20892 USA. NR 50 TC 7 Z9 7 U1 0 U2 0 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 SEP 16 PY 1996 VL 733 IS 2 BP 203 EP 210 DI 10.1016/0006-8993(96)00535-5 PG 8 WC Neurosciences SC Neurosciences & Neurology GA VK400 UT WOS:A1996VK40000007 PM 8891303 ER PT J AU Maurice, T Roman, FJ Su, TP Privat, A AF Maurice, T Roman, FJ Su, TP Privat, A TI Beneficial effects of sigma agonists on the age-related learning impairment in the senescence-accelerated mouse (SAM) SO BRAIN RESEARCH LA English DT Article DE sigma(a) agonists; JO-1784 (igmesine); PRE-084; senescence-accelerated mouse (SAM); learning and memory; aging; passive avoidance; Y-maze; water-maze ID INDUCED NEURONAL ACTIVATION; BEHAVIORAL EVIDENCE; SELECTIVE LIGAND; MEMORY PROCESSES; MODULATING ROLE; BRAIN-SLICES; H-3 TCP; MICE; RECEPTORS; RETENTION AB A beneficial effect of sigma (sigma) agonists was previously described on several pharmacological models of learning impairments. We examined this effect in senescence-accelerated mice (SAM), which has been developed as a murine model of aging and cognitive dysfunction. SAMP8/Ta (P8, senescence-prone substrain), 10-12 months of age, showed significant impairments in mnemonic capacities, as compared to age-matched SAMR1/Ta controls (R1, senescence-resistant substrain). Tests included open-field behavior, spontaneous alternation performances in the Y-maze, step-down passive avoidance and place learning after repetitive training in a water-maze. Pretreatment with the sigma agonists JO-1784 (igmesine) or PRE-084, at 0.1-3 mg/kg, s.c., significantly improved spontaneous alternation and passive avoidance performances in P8. JO-1784 or PRE-084, at 1 mg/kg, also improved place learning in the water-maze, and retention, in term of escape latency. The implication of sigma sites was indicated by the lack of significant effect of JO-1783, the inactive enantiomer of JO-1784, and by the ability of BMY-14802 (5 mg/kg, i.p.) to antagonize the effects on passive avoidance of JO-1784 (0.5 mg/kg) or PRE-084 (1 mg/kg). Subchronic treatments with JO-1784 (0.5 mg/kg/day) or PRE-084 (1 mg/kg/day) during 10 days, allowed a significant improvement of learning during training in the water-maze, but retention was not significantly ameliorated. These results confirmed the interest of the SAM substrains as an experimental model for senile memory impairment and showed that sigma agonists could improve the quality of learning, although they seem less effective on long-term memory retrieval upon chronic administration. C1 INST RECH JOUVEINAL,DEPT BIOCHIM,F-94263 FRESNES,FRANCE. NIDA,INTRAMURAL RES PROGRAM,BALTIMORE,MD 21224. RP Maurice, T (reprint author), ECOLE NATL SUPER CHIM MONTPELLIER,INSERM U336,8 RUE ECOLE NORMALE,F-34053 MONTPELLIER 01,FRANCE. NR 37 TC 42 Z9 44 U1 1 U2 7 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0006-8993 J9 BRAIN RES JI Brain Res. PD SEP 16 PY 1996 VL 733 IS 2 BP 219 EP 230 DI 10.1016/0006-8993(96)00565-3 PG 12 WC Neurosciences SC Neurosciences & Neurology GA VK400 UT WOS:A1996VK40000009 PM 8891305 ER PT J AU Blount, P Sukharev, SI Moe, PC Schroeder, MJ Guy, HR Kung, C AF Blount, P Sukharev, SI Moe, PC Schroeder, MJ Guy, HR Kung, C TI Membrane topology and multimeric structure of a mechanosensitive channel protein of Escherichia coli SO EMBO JOURNAL LA English DT Article DE mechanosensation; mechanosensitive channel; MscL; osmotic forces; stretch activated channel ID EPITHELIAL SODIUM-CHANNEL; PATCH-CLAMP; ION CHANNELS; FUNCTIONAL RECONSTITUTION; ACTIVATED CHANNELS; POTASSIUM CHANNEL; BACILLUS-SUBTILIS; GENE; CLONING; MSCL AB We have studied the membrane topology and multimeric structure of a mechanosensitive channel, MscL, which we previously isolated and cloned from Escherichia coli. We have localized this 15-kDa protein to the inner membrane and, by PhoA fusion, have shown that it contains two transmembrane domains with both the amino and carboxyl termini on the cytoplasmic side, Mutation of the glutamate at position 56 to histidine led to changes in channel kinetics which were dependent upon the pH on the periplasmic, but not cytoplasmic side of the membrane, providing additional evidence for the periplasmic positioning of this part of the molecule, Tandems of two MscL subunits expressed as a single polypeptide formed functional channels, suggesting an even number of transmembrane domains per subunit (amino and carboxyl termini on the same side of the membrane), and an even number of subunits per functional complex, Finally, cross-linking studies suggest that the functional MscL complex is a homo-hexamer. In summary, these data are all consistent with a protein domain assignment and topological model which we propose and discuss. C1 UNIV WISCONSIN, MOL BIOL LAB, MADISON, WI 53706 USA. UNIV WISCONSIN, DEPT GENET, MADISON, WI 53706 USA. NCI, MATH BIOL LAB, NIH, BETHESDA, MD 20892 USA. FU NIGMS NIH HHS [GM47856] NR 53 TC 131 Z9 137 U1 2 U2 5 PU NATURE PUBLISHING GROUP PI NEW YORK PA 75 VARICK ST, 9TH FLR, NEW YORK, NY 10013-1917 USA SN 0261-4189 J9 EMBO J JI Embo J. PD SEP 16 PY 1996 VL 15 IS 18 BP 4798 EP 4805 PG 8 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA VL095 UT WOS:A1996VL09500003 PM 8890153 ER PT J AU Tagaya, Y Burton, JD Miyamoto, Y Waldmann, TA AF Tagaya, Y Burton, JD Miyamoto, Y Waldmann, TA TI Identification of a novel receptor signal transduction pathway for IL-15/T in mast cells SO EMBO JOURNAL LA English DT Article DE interleukin-2 receptor; interleukin-15; JAK kinases; mast cells ID PROTEIN-TYROSINE KINASES; ACTIVATED KILLER-CELLS; JAK-3 JANUS KINASE; IL-2 RECEPTOR; BETA-CHAIN; INTERLEUKIN-2 RECEPTOR; ALPHA-CHAIN; ERYTHROPOIETIN RECEPTOR; STAT PROTEINS; GAMMA-CHAIN AB Interleukin-15/T(IL-15) is a growth factor that utilizes IL-2 receptor (IL-2R) components in addition to its private binding protein IL-15R alpha in T-cells. Here, we report that IL-15 induces mast cell proliferation in the absence of IL-2R alpha and beta. Using transfectants of these cells with a cytoplasmic-truncated mutant of gamma c. we demonstrated that IL-15 signaling in mast cells does not involve gamma c. Cross-linking of mast cells with [I-125]IL-15 revealed a 60-65 kDa IL-15 binding protein that is distinct from known components of T-cell IL-15 receptors. Mast cell IL-15 receptors recruit JAK-2 and STAT-5, instead of JAK1/3 and STAT3/5 that are activated in T-cells. Thus IL-15 is a mast cell growth factor that utilizes a novel receptor and distinct signaling pathway. RP Tagaya, Y (reprint author), NCI,METAB BRANCH,NATL INST HLTH,9000 ROCKVILLE PIKE,BETHESDA,MD 20892, USA. NR 38 TC 127 Z9 130 U1 1 U2 2 PU OXFORD UNIV PRESS UNITED KINGDOM PI OXFORD PA WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP SN 0261-4189 J9 EMBO J JI Embo J. PD SEP 16 PY 1996 VL 15 IS 18 BP 4928 EP 4939 PG 12 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA VL095 UT WOS:A1996VL09500016 PM 8890166 ER PT J AU Ura, K Nightingale, K Wolffe, AP AF Ura, K Nightingale, K Wolffe, AP TI Differential association of HMG1 and linker histones B4 and H1 with dinucleosomal DNA: Structural transitions and transcriptional repression SO EMBO JOURNAL LA English DT Article DE dinucleosomes; DNA; histone B4; histone H1; HMG1 ID EARLY XENOPUS EMBRYOGENESIS; RNA-POLYMERASE TRANSCRIBES; MOBILITY GROUP PROTEIN-1; GENE-TRANSCRIPTION; CHROMOSOMAL-PROTEINS; NUCLEOSOME MOBILITY; GLOBULAR DOMAIN; CORE HISTONES; IN-VIVO; CHROMATIN AB We examined the structural and functional consequences of incorporating either histone H1, histone B4 or HMG1 into a synthetic dinucleosome containing two 5S rRNA genes, We found that all three proteins bind to linker DNA, stabilizing an additional 20 bp from micrococcal nuclease digestion and restrict nucleosome mobility, Histone H1 has the highest-affinity interaction with the dinucleosome; histone B4 and HMG1 associate with significantly reduced affinities, We found that histone H1 binds to the dinucleosome template with a dissociation constant (K-D) of 7.4 nM, whereas the K-D is 45 nM for histone B4 and 300 nM for HMG1. The K(D)s for the interaction of these proteins with naked DNA are 18 nM for H1, 80 nM for B4 and 300 nM for HMG1, The differences in association of these proteins with the dinucleosome are reflected in the efficiency with which the different proteins repress transcription from the 5S rRNA genes, Thus, although all three proteins can contribute to the organization of chromatin, the stability of the structures they assemble will vary, Our results provide a molecular explanation for the transcriptional promiscuity of Xenopus early embryonic chromatin, which is enriched in HMG1 and linker histone B4, but deficient in histone H1. RP Ura, K (reprint author), NICHHD,MOL EMBRYOL LAB,NIH,BLDG 6,ROOM B1A-13,BETHESDA,MD 20892, USA. NR 80 TC 99 Z9 100 U1 0 U2 1 PU OXFORD UNIV PRESS UNITED KINGDOM PI OXFORD PA WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP SN 0261-4189 J9 EMBO J JI Embo J. PD SEP 16 PY 1996 VL 15 IS 18 BP 4959 EP 4969 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA VL095 UT WOS:A1996VL09500019 PM 8890169 ER PT J AU Altshuller, Y Copeland, NG Gilbert, DJ Jenkins, NA Frohman, MA AF Altshuller, Y Copeland, NG Gilbert, DJ Jenkins, NA Frohman, MA TI Gcm1, a mammalian homolog of Drosophila glial cells missing SO FEBS LETTERS LA English DT Article DE Gcm; glia; glial cells missing; gliogenesis; mGCM1; neurogenesis ID SEQUENCES; SWITCH AB Differentiation of glia (astrocytes and oligodendrocytes) in Drosophila requires the gene glial cells missing (gcm), which controls lineage determination, In the absence of gcm, neuroglia progenitors exclusively differentiate into neurons, instead of into both neurons and glia, In contrast, ectopic overexpression of gcm causes uniform differentiation of the neuroglia progenitors into glia, Glial and neuronal cells in vertebrates similarly derive from neuroblast progenitors, To investigate vertebrate glial formation, we have identified, cloned, and chromosomally mapped a mammalian gcm homolog. Mouse Gcm1 demonstrates extensive similarity to Drosophila gcm but is expressed at very low levels during neuro- and gliogenesis. C1 SUNY STONY BROOK,DEPT PHARMACOL SCI,STONY BROOK,NY 11794. SUNY STONY BROOK,INST CELL & MOL BIOL,STONY BROOK,NY 11794. NCI,FREDERICK CANC RES & DEV CTR,ABL BASIC RES PROGRAM,MAMMALIAN GENET LAB,FREDERICK,MD 21702. FU NICHD NIH HHS [HD29758] NR 14 TC 55 Z9 56 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 SEP 16 PY 1996 VL 393 IS 2-3 BP 201 EP 204 DI 10.1016/0014-5793(96)00890-3 PG 4 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA VH654 UT WOS:A1996VH65400012 PM 8814290 ER PT J AU Felder, CC Nielsen, A Briley, EM Palkovits, M Priller, J Axelrod, J Nguyen, DN Richardson, JM Riggin, RM Koppel, GA Paul, SM Becker, GW AF Felder, CC Nielsen, A Briley, EM Palkovits, M Priller, J Axelrod, J Nguyen, DN Richardson, JM Riggin, RM Koppel, GA Paul, SM Becker, GW TI Isolation and measurement of the endogenous cannabinoid receptor agonist, anandamide, in brain and peripheral tissues of human and rat SO FEBS LETTERS LA English DT Article DE anandamide; cannabis; cannabinoid receptor; marijuana ID PHARMACOLOGICAL ACTIVITY; REGIONAL DISTRIBUTION; SIGNAL-TRANSDUCTION; LIGAND; CONSTITUENT; EXPRESSION; BINDS; CB1 AB Anandamide (arachidonylethanolamide) is a novel lipid neurotransmitter first isolated from porcine brain which has been shown to be a functional agonist for the cannabinoid CB1 and CB2 receptors, Anandamide has never been isolated from human brain or peripheral tissues and its role in human physiology has not been examined, Anandamide was measured by LC/MS/MS and was found in human and rat hippocampus (and human parahippocampal cortex), striatum, and cerebellum, brain areas known to express high levels of CB1 cannabinoid receptors, Significant levels of anandamide were also found in the thalamus which expresses low levels of CB1 receptors, Anandamide was also found in human and rat spleen which expresses high levels of the CB2 cannabinoid receptor, Small amounts of anandamide were also detected in human heart and rat skin, Only trace quantities were detected in pooled human serum, plasma, and CSF, The distribution of anandamide in human brain and spleen supports its potential role as an endogenous agonist in central and peripheral tissues, The low levels found in serum, plasma, and CSF suggest that it is metabolized in tissues where it is synthesized, and that its action is probably not hormonal in nature. C1 ELI LILLY & CO,LILLY RES LAB,INDIANAPOLIS,IN 46285. RP Felder, CC (reprint author), NIMH,CELL BIOL LAB,BLDG 36,RM 3A-15,36 CONVENT DR MSC 4090,BETHESDA,MD 20892, USA. RI Palkovits, Miklos/F-2707-2013; OI Palkovits, Miklos/0000-0003-0578-0387 NR 23 TC 235 Z9 242 U1 2 U2 11 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0014-5793 J9 FEBS LETT JI FEBS Lett. PD SEP 16 PY 1996 VL 393 IS 2-3 BP 231 EP 235 DI 10.1016/0014-5793(96)00891-5 PG 5 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA VH654 UT WOS:A1996VH65400018 PM 8814296 ER PT J AU Huang, XL Smith, MC Berzofsky, JA Barchi, JJ AF Huang, XL Smith, MC Berzofsky, JA Barchi, JJ TI Structural comparison of a 15 residue peptide from the V3 loop of HIV-1(IIIb) and an O-glycosylated analogue SO FEBS LETTERS LA English DT Article DE NMR; V3 loop; glycopeptide; CTL ID HUMAN-IMMUNODEFICIENCY-VIRUS; PRINCIPAL NEUTRALIZING DETERMINANT; TYPE-1 ENVELOPE GLYCOPROTEIN; CLASS-I; MONOCLONAL-ANTIBODIES; MHC MOLECULE; GP120; PROTEIN; SPECTROSCOPY; GP160 AB As part of a program to study the effect of glycosylation an the three-dimensional structures of HIV-1(IIIB) V3 peptide constructs, we have examined the solution structures of a 15 residue peptide (RIQRGPGRAFVTIGK, P18(IIIB)), originally mapped as an epitope recognized by CD8(+) D-d class I MHC-restricted murine cytotoxic T-lymphocytes (CTL), and an analogue (P18(IIIB)-g), O-glycosylated with an alpha-galactosamine on Thr-12, using NMR, circular dichroism and molecular modeling methods, Our studies show Chat the peptides sample mainly random conformations in aqueous solution near 25 degrees C and become more ordered by the addition of trifluoroethanol, Upon decreasing the temperature to 5 degrees C, a reverse turn is famed around the immunodominant tip (G(5)-R(8)). Glycosylation on T-12 'tightens' the turn slightly as suggested by NOE and CD analysis, Ln addition, the sugar has a defined conformation with respect to the peptide backbone and influences the local peptide conformation, These data suggest that simple glycosylation mag. influence the conformational equilibrium of a V3 peptide which contains a domain critical for antibody recognition and virus neutralization, We also show that the ability of cytotoxic T-lymphocytes (CTL) to lyse tumor cells presenting P18(IIIB) was completely abrogated by threonine glycosylation. C1 NCI,MED CHEM LAB,DIV BASIC SCI,NIH,BETHESDA,MD 20892. NCI,MOL IMMUNOGENET & VACCINE RES SECT,DIV BASIC SCI,NIH,BETHESDA,MD 20892. RI Barchi Jr., Joseph/N-3784-2014 NR 44 TC 29 Z9 29 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0014-5793 J9 FEBS LETT JI FEBS Lett. PD SEP 16 PY 1996 VL 393 IS 2-3 BP 280 EP 286 DI 10.1016/0014-5793(96)00912-X PG 7 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA VH654 UT WOS:A1996VH65400027 PM 8814305 ER PT J AU Muntaner, C Nieto, FJ OCampo, P AF Muntaner, C Nieto, FJ OCampo, P TI The bell curve: On race, social class, and epidemiologic research SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Review ID BIRTH-WEIGHT; PERINATAL-MORTALITY; INFANT-MORTALITY; AFRICAN-AMERICAN; BLOOD-PRESSURE; RISK-FACTORS; HEALTH; ORIGIN; DISCRIMINATION; NEUTROPENIA C1 NIMH,NIH,BETHESDA,MD 20892. JOHNS HOPKINS UNIV,SCH HYG & PUBL HLTH,DEPT EPIDEMIOL,BALTIMORE,MD. JOHNS HOPKINS UNIV,SCH HYG & PUBL HLTH,DEPT MATERNAL & CHILD HLTH,BALTIMORE,MD. RP Muntaner, C (reprint author), W VIRGINIA SCH MED,INST OCCUPAT & ENVIRONM HLTH,3313 ROBERT C BYRD HLTH SCI CTR,POB 9190,MORGANTOWN,WV 26506, USA. RI Muntaner, C/A-5043-2010 NR 75 TC 69 Z9 73 U1 0 U2 3 PU AMER J EPIDEMIOLOGY PI BALTIMORE PA 624 N BROADWAY RM 225, BALTIMORE, MD 21205 SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD SEP 15 PY 1996 VL 144 IS 6 BP 531 EP 536 PG 6 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA VG930 UT WOS:A1996VG93000001 PM 8797512 ER PT J AU Zhang, XK Dutky, RC Fales, HM AF Zhang, XK Dutky, RC Fales, HM TI Rubber stoppers as sources of contaminants in electrospray analysis of peptides and proteins SO ANALYTICAL CHEMISTRY LA English DT Article ID DRUGS; EXTRACTION; BINDING; BLOOD AB Using MS/MS, we have identified ions at m/z 399, 421, 609.4, and 819.9 as those of tris(2-butoxyethyl) phosphate, originating in the gray rubber stoppers commonly used as closures in commercial sources of peptides and proteins. The compound is readily extracted by contact with organic solvents such as alcohol or acetic acid and even by water alone, Zinc salts of proteins are also observed, presumably from zinc oxide fillers present in the stoppers. C1 NHLBI,CHEM STRUCT SECT,BIOPHYS CHEM LAB,NIH,BETHESDA,MD 20892. NR 19 TC 12 Z9 13 U1 0 U2 5 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 SN 0003-2700 J9 ANAL CHEM JI Anal. Chem. PD SEP 15 PY 1996 VL 68 IS 18 BP 3288 EP 3289 DI 10.1021/ac960245n PG 2 WC Chemistry, Analytical SC Chemistry GA VG141 UT WOS:A1996VG14100033 PM 8797388 ER PT J AU VanderPutten, DM Torrey, EF Larive, AB Merril, CR AF VanderPutten, DM Torrey, EF Larive, AB Merril, CR TI Plasma protein variations in monozygotic twins discordant for schizophrenia SO BIOLOGICAL PSYCHIATRY LA English DT Article DE schizophrenia; protein; twins; plasma; haptoglobin ID 2-DIMENSIONAL ELECTROPHORESIS; CEREBROSPINAL-FLUID; POLYPEPTIDES; DISEASE AB Monozygotic twins discordant for schizophrenia were analyzed by two-dimensional (2-D) gel electrophoresis to identify extrahereditary factors important in the development of schizophrenia. Plasma protein patterns in 2-D gels of monozygotic twins discordant for schizophrenia were found to be significantly less alike than those of normal control monozygotic twins. Several polypeptide spots were found to be elevated in the plasma of the schizophrenic twin. One of these polypeptides, spot 782, was also found to be significantly (p <.001) elevated when schizophrenic patients were compared to unrelated normal control individuals. Spot 782 may be an isoform of haptoglobin. Quantitative variations in some plasma haptoglobin levels were seen between discordant twins, but not between unrelated schizophrenic and normal control individuals. C1 MONOCLONET INT INC,HOUSTON,TX. RP VanderPutten, DM (reprint author), ST ELIZABETH HOSP,NIMH,NEUROSCI RES CTR,ROOM 12 WAW BLDG,WASHINGTON,DC 20032, USA. NR 15 TC 2 Z9 2 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 SN 0006-3223 J9 BIOL PSYCHIAT JI Biol. Psychiatry PD SEP 15 PY 1996 VL 40 IS 6 BP 437 EP 442 DI 10.1016/0006-3223(95)00467-X PG 6 WC Neurosciences; Psychiatry SC Neurosciences & Neurology; Psychiatry GA VG789 UT WOS:A1996VG78900001 PM 8879462 ER PT J AU LeBeau, MM Espinosa, R Davis, EM Eisenbart, JD Larson, RA Green, ED AF LeBeau, MM Espinosa, R Davis, EM Eisenbart, JD Larson, RA Green, ED TI Cytogenetic and molecular delineation of a region of chromosome 7 commonly deleted in malignant myeloid diseases SO BLOOD LA English DT Article ID YEAST ARTIFICIAL CHROMOSOME; IN-SITU HYBRIDIZATION; LONG-ARM DELETIONS; HUMAN CANCER; LEUKEMIA; DISORDERS; HUMAN-CHROMOSOME-7; IDENTIFICATION; ABNORMALITIES; BREAKPOINTS AB Loss of a whole chromosome 7 or a deletion of the long arm, del(7q), are recurring abnormalities in malignant myeloid diseases. To determine the location of genes on 7q that are likely to play a role in leukemogenesis, we examined the deleted chromosome 7 homologs in a series of 81 patients with therapy-related or de novo myelodysplastic syndrome or acute myeloid leukemia. Our analysis showed that the deletions were interstitial and that there were two distinct deleted segments of 7q. The majority of patients (65 of 81 [80%]) had proximal breakpoints in hands q11-22 and distal breakpoints in q31-36; the smallest overlapping deleted segment was within q22. The remaining 16 patients had deletions involving the distal q arm with a commonly deleted segment of q32-33. To define the proximal deleted segment at 7q22 at a molecular level, we used fluorescence in situ hybridization with a panel of mapped yeast artificial chromosome (YAC) clones from 7q to examine 15 patients with deletion breakpoints in 7q22, Wa determined that the smallest overlapping deleted segment is contained in a well-defined YAC contig that spans 2 to 3 Mb. These studies delineate the region of 7q that must be searched to isolate a putative myeloid leukemia suppressor gene, and provide the necessary cloned DNA for more detailed physical mapping and gene isolation. (C) 1996 by The American Society of Hematology. C1 UNIV CHICAGO,CANC RES CTR,CHICAGO,IL 60637. NIH,GENOME TECHNOL BRANCH,NATL CTR HUMAN GENOME RES,BETHESDA,MD. RP LeBeau, MM (reprint author), UNIV CHICAGO,HEMATOL ONCOL SECT,5841 S MARYLAND AVE,MC2115,CHICAGO,IL 60637, USA. OI Larson, Richard/0000-0001-9168-3203 FU NCI NIH HHS [P01 CA40046] NR 39 TC 165 Z9 168 U1 1 U2 1 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0006-4971 J9 BLOOD JI Blood PD SEP 15 PY 1996 VL 88 IS 6 BP 1930 EP 1935 PG 6 WC Hematology SC Hematology GA VK565 UT WOS:A1996VK56500002 PM 8822909 ER PT J AU Maeno, M Mead, PE Kelley, C Xu, RH Kung, HF Suzuki, A Ueno, N Zon, LI AF Maeno, M Mead, PE Kelley, C Xu, RH Kung, HF Suzuki, A Ueno, N Zon, LI TI The role of BMP-4 and GATA-2 in the induction and differentiation of hematopoietic mesoderm in Xenopus laevis SO BLOOD LA English DT Article ID BONE MORPHOGENETIC PROTEIN-4; GLOBIN GENE-EXPRESSION; ACTIVIN-A; VENTRAL MESODERM; EMBRYONIC-DEVELOPMENT; BLOOD; ERYTHROPOIESIS; PROLIFERATION; PROGENITORS; MODULATION AB Vertebrate embryonic blood formation is regulated by factors that participate in dorsal-ventral patterning and mesoderm induction. The GATA-binding transcription factors are required for normal hematopoiesis and are expressed during gastrulation when ventral mesoderm (VM) is induced to form blood. Based on the recent demonstration that bone morphogenetic protein (BMP-4) is a potent ventralizing factor and inducer of hematopoietic tissue, we hypothesized that GATA-2 could be induced or activated by BMP-4. Here we demonstrate that BMP-4 can stimulate GATA-5 expression, and that expression of a dominant negative BMP-4 receptor can suppress GATA-2 induction by BMP-4 in ventral mesoderm, Over-expression of GATA-2 in ventral mesoderm leads to increased globin production and forced expression of GATA-2 in primitive ectoderm adjacent to ventral mesoderm also stimulates globin expression. Our results suggest that BMP-4 and GATA-2 can function in two adjacent germ layers, mesoderm and ectoderm, to participate in blood cell formation during embryogenesis. (C) 1996 by The American Society of Hematology. C1 CHILDRENS HOSP,HOWARD HUGHES MED INST,DEPT PEDIAT,DIV HEMATOL,BOSTON,MA 02115. NIIGATA UNIV,FAC SCI,DEPT BIOL,NIIGATA 95021,JAPAN. NCI,FREDERICK CANC RES & DEV CTR,DEPT BASIC SCI,LAB BIOCHEM PHYSIOL,FREDERICK,MD 21701. HOKKAIDO UNIV,FAC PHARMACEUT SCI,SAPPORO,HOKKAIDO 060,JAPAN. RI Kelley, Clair/O-2688-2015; Xu, Ren-He/M-3125-2016 FU NHLBI NIH HHS [R01-HL48801] NR 37 TC 146 Z9 151 U1 0 U2 2 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0006-4971 J9 BLOOD JI Blood PD SEP 15 PY 1996 VL 88 IS 6 BP 1965 EP 1972 PG 8 WC Hematology SC Hematology GA VK565 UT WOS:A1996VK56500008 PM 8822915 ER PT J AU Maciejewski, JP Selleri, C Sato, T Anderson, S Young, NS AF Maciejewski, JP Selleri, C Sato, T Anderson, S Young, NS TI A severe and consistent deficit in marrow and circulating primitive hematopoietic cells (long-term culture-initiating cells) in acquired aplastic anemia SO BLOOD LA English DT Article ID BLOOD STEM-CELLS; LIMITING DILUTION; PERIPHERAL-BLOOD; PROGENITOR CELLS; BONE-MARROW; CORD-BLOOD; PURIFICATION AB We examined the stem cell compartment of patients with acquired aplastic anemia (AA) using the long-term culture-initiating cell assay (LTC-IC), in parallel with measurements of CD34(+) cells and mature hematopoietic progenitors. Secondary colonies from cells surviving 5 weeks of long-term bone marrow culture (LTBMC) were determined for the peripheral blood (PB) of 68 AA patients and 13 normal controls and for BM of 49 AA patients and 14 controls; because of low cell numbers, formal limiting dilution analysis could only be performed in 10 patients. The relationship of cell input in LTBMC and the output of secondary colonies was linear, allowing quantification of LTC-IC number from bulk cultures. Secondary colony formation was markedly abnormal in severe AA. In contrast to 7.8 colony-forming cells (CFC)/10(5) mononuclear cells in normal BM and 0.14 CFC/10(5) normal PB mononuclear cells, patients with severe disease showed 0.024 CFC/10(5) in BM and 0.0068 CFC/10(5) in PB. Under limiting dilution conditions, patients' cells also showed markedly lower colony-forming ability. In contrast to 4.3 +/- 1 colonies/normal LTC-IC, we obtained only 1.27 +/- 0.09 and 2.0 +/- 0.35 colonies from BM of acute and recovered cases, respectively. These values were used to extrapolate LTC-IC numbers from secondary colony formation in suspension cultures. In PB, calculated LTC-IC were decreased 7.4-fold in new and relapsed severe AA and 2.8-fold in recovered AA. In BM, LTC-IC were decreased 10-fold in new and relapsed AA and sixfold in recovered cases. Compared with measurements obtained on presentation, LTC-IC were lower in posttreatment samples from patients who had failed to recover after intensive immunosuppression and relatively higher in cases at relapse. In recovered patients, LTC-IC number increased but remained below the normal range in 20 of 25. In patients studied serially for 3 to 12 months after treatment, LTC-IC numbers remained stable but low. LTC-IC number correlated with concurrently determined CD34(+) cell number and primary hematopoietic colony formation. These results indicate that stem cell numbers, as quantitated by the LTC-IC assay, are markedly diminished in number in all severe AA. Additionally, the function of the stem cell or the stem cell compartment in AA is also abnormal, as inferred from the low clonogenic potential in secondary colony assays. Early hematologic improvement in some patients occurs without increasing numbers of LTC-IC, and a minority of recovered cases show apparent repopulation of the LTC-IC compartment years after treatment. (C) 1996 by The American Society of Hematology. RP Maciejewski, JP (reprint author), NHLBI,HEMATOL BRANCH,BLDG 10,ROOM 7C103,BETHESDA,MD 20892, USA. NR 23 TC 85 Z9 86 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0006-4971 J9 BLOOD JI Blood PD SEP 15 PY 1996 VL 88 IS 6 BP 1983 EP 1991 PG 9 WC Hematology SC Hematology GA VK565 UT WOS:A1996VK56500010 PM 8822917 ER PT J AU Vallera, DA PanoskaltsisMortari, A Jost, C Ramakrishnan, S Eide, CR Kreitman, RJ Nicholls, PJ Pennell, C Blazar, BR AF Vallera, DA PanoskaltsisMortari, A Jost, C Ramakrishnan, S Eide, CR Kreitman, RJ Nicholls, PJ Pennell, C Blazar, BR TI Anti-graft-versus-host disease effect of DT390-anti-CD3sFv, a single-chain Fv fusion immunotoxin specifically targeting the CD3 epsilon moiety of the T-Cell receptor SO BLOOD LA English DT Article ID RICIN-A-CHAIN; PHASE-I TRIAL; BONE-MARROW TRANSPLANTATION; DIPHTHERIA-TOXIN; MONOCLONAL-ANTIBODIES; HISTOCOMPATIBILITY BARRIER; HEMATOLOGIC MALIGNANCIES; MICE; PROPHYLAXIS; EXPRESSION AB In a recent study, we showed that an immunotoxin (IT) made with a conventional monoclonal antibody targeting the CD3 epsilon moiety of the T-cell receptor (ICR) had a potent, but partial, graft-versus-host disease (GVHD) effect (Vallera et al, Blood 86:4367, 1995), Therefore, in this current study, we determined whether a fusion immunotoxin made with anti-CD3 single-chain Fv (sFv), the smallest unit of antibody recognizing antigen, would have anti-GVHD activity. A fusion protein was synthesized from a construct made by splicing sFv cDNA from the hybridoma 145-2C11 to a truncated form of the diphtheria toxin (DT390) gene. DT390 encodes a molecule that retains full enzymatic activity, but excludes the native DT binding domain, The DT390-anti-CD3sFv hybrid gene was cloned into a vector under the control of an inducible promoter. The protein was expressed in Escherichia coli and then purified from inclusion bodies. The DT390 moiety of the protein had full enzymatic activity compared with native DT and DT390-anti-CD3sFv, with an IC50 of 1 to 2 nmol/L against phytohemagglutinin-stimulated and alloantigen-stimulated T cells, Specificity was shown (1) by blocking the IT with parental anti-CD3 antibody, but not with a control antibody; (2) by failure of DT390-anti-CD3sFv to inhibit lipopolysaccharide-stimulated murine B cells; (3) by failure of an Ig control fusion protein, DT390-Fc, to inhibit T-cell responses; and (4) with in vivo immunohistochemisty studies. GVHD was studied in a model in which C57BL/6 (H-2(b))-purified lymph node T cells were administered to major histocompatibility complex (MHC) antigen disparate unirradiated C.B.-17 scid (H-2(d)) mice to assess GVHD effects in the absence of irradiation toxicity, Flow cytometry studies showed that donor T cells were expanded 57-fold and histopathologic analysis showed the hallmarks of a lethal model of GVHD. Control mice receiving phosphate-buffered saline showed 17% survival on day 80 after bone marrow transplantation, and mice receiving 2 mu g DT390-Fc fusion toxin control administered in 2 daily doses for 6 days (days 0 through 5) had a 43% survival rate. In contrast, 86% of mice receiving the same dose of DT390-anti-CD3sFv were survivors on day 80, a significant improvement, although survivors still showed histopathologic signs of GVHD. These findings suggest that new anti-GVHD agents can be genetically engineered and warrant further investigation of fusion proteins for GVHD treatment. (C) 1996 by The American Society of Hematology. C1 UNIV MINNESOTA HOSP & CLIN,SECT EXPT CANC IMMUNOL,MINNEAPOLIS,MN 55455. UNIV MINNESOTA HOSP & CLIN,DEPT LAB MED,MINNEAPOLIS,MN 55455. UNIV MINNESOTA HOSP & CLIN,DEPT PHARMACOL,MINNEAPOLIS,MN 55455. UNIV MINNESOTA HOSP & CLIN,DEPT PEDIAT,MINNEAPOLIS,MN 55455. UNIV MINNESOTA HOSP & CLIN,DIV BONE MARROW TRANSPLANTAT,MINNEAPOLIS,MN 55455. NCI,NATL INST HLTH,BETHESDA,MD 20892. RP Vallera, DA (reprint author), UNIV MINNESOTA HOSP & CLIN,DEPT THERAPEUT RADIOL,BOX 367,HARVARD ST & E RIVER RD,MINNEAPOLIS,MN 55455, USA. FU NCI NIH HHS [R01-CA36725, P01-CA21737]; NIAID NIH HHS [R01-AI34495] NR 56 TC 24 Z9 24 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 SN 0006-4971 J9 BLOOD JI Blood PD SEP 15 PY 1996 VL 88 IS 6 BP 2342 EP 2353 PG 12 WC Hematology SC Hematology GA VK565 UT WOS:A1996VK56500050 PM 8822957 ER PT J AU Taylor, JA Hirvonen, A Watson, M Pittman, G Mohler, JL Bell, DA AF Taylor, JA Hirvonen, A Watson, M Pittman, G Mohler, JL Bell, DA TI Association of prostate cancer with vitamin D receptor gene polymorphism SO CANCER RESEARCH LA English DT Article ID CELL-PROLIFERATION; BONE-DENSITY; ALLELES; WOMEN; BSMI AB The incidence of prostate cancer in the United States is second only to skin cancers, and the disease kills almost the same number of men as breast cancer does women, Relatively few risk factors are known for prostate cancer, although several lines of evidence suggest that vitamin D may be an important determinant of prostate cancer risk, A series of common polymorphisms in the vitamin D receptor gene were recently reported to be associated with bone density and risk of osteoporosis (Morrison et al., Nature (Lond.), 367: 284-287, 1994), These genetic variants have been correlated with both circulating levels of active vitamin D hormone and in vitro measures of gene expression (Morrison et al., Nature (Lond.), 367: 284-287, 1994). We tested the hypothesis that vitamin D receptor gene polymorphisms are associated with prostate cancer risk using a case-control study of 108 men undergoing radical prostatectomy and 170 male urology clinic controls with no history of cancer, Among the white control group, 22% were homozygous for the presence of a TaqI RFLP at codon 352 (genotype tt), but only 8% of cases had this genotype (P < 0.01), A similar trend mas seen among the small number of blacks in this study (13% for controls, 8% for cases), although the difference was not statistically significant, Race-adjusted combined analysis suggests that men who are homozygous for the t allele (shown to correlate with higher serum levels of the active form of vitamin D) have one-third the risk of developing prostate cancer requiring prostatectomy compared to men who are heterozygotes or homozygous for the T allele (odds ratio(MH) = 0.34; 95% confidence interval, 0.16-0.76; P < 0.01), These results support recent ecological, population, and in vitro studies suggesting that vitamin D is an important determinant of prostate cancer risk and, if confirmed, suggest strategies for chemoprevention of this common cancer. C1 NIEHS,BIOCHEM RISK ANAL LAB,RES TRIANGLE PK,NC 27709. UNIV N CAROLINA,CHAPEL HILL,NC 27599. RP Taylor, JA (reprint author), NIEHS,EPIDEMIOL BRANCH,MAIL DROP A3-05,POB 12233,RES TRIANGLE PK,NC 27709, USA. OI taylor, jack/0000-0001-5303-6398 NR 18 TC 243 Z9 257 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 0008-5472 J9 CANCER RES JI Cancer Res. PD SEP 15 PY 1996 VL 56 IS 18 BP 4108 EP 4110 PG 3 WC Oncology SC Oncology GA VG370 UT WOS:A1996VG37000005 PM 8797574 ER PT J AU Vasandani, VM Wu, YN Mikulski, SM Youle, RJ Sung, C AF Vasandani, VM Wu, YN Mikulski, SM Youle, RJ Sung, C TI Molecular determinants in the plasma clearance and tissue distribution of ribonucleases of the ribonuclease A superfamily SO CANCER RESEARCH LA English DT Article ID P-30 PROTEIN; CYTOTOXIC RIBONUCLEASES; SOLID TUMORS; INHIBITION; ANGIOGENIN; EXPRESSION; CARCINOMA; CHIMERAS; KINETICS; EMBRYOS AB The similarities and differences among members of the RNase A superfamily provide an ideal opportunity to examine the molecular basis for differences in their pharmacokinetics and biodistribution. Plasma clearances in BALB/c mice are similar among the five RNases studied: human pancreatic RNase, angiogenin, eosinophil-derived neurotoxin, onconase, and bovine seminal RNase. The average clearance is 0.13 ml/min or 60% of the glomerular filtration rate (measured by [C-14]inulin clearance during continuous infusion from an i.p. implanted osmotic pump). Angiogenin has a higher volume of distribution and plasma-to-muscle transport rate than the other RNases, suggestive of binding to endothelial cells. Organ distribution differs dramatically among these RNases. The RNase most toxic to tumor cells, onconase, exhibits the longest retention in the kidneys: at 180 min, 50% of the injected dose is found in the kidneys, whereas only 1% or less of the other RNases is retained in the kidneys. Slower elimination of onconase from the kidneys may be due to a higher degree of binding in the kidney or a resistance to proteolytic degradation. To elucidate the molecular determinants involved in tissue uptake, we examined the biodistribution of recombinant onconase and two onconase-pancreatic RNase chimeric proteins. The tissue retention property of onconase appears to be located in at least two regions, one of which is in the NH2-terminal 9-amino acid alpha-helix. The NH2-terminal pyroglutamate of onconase, a residue essential for ribonucleolytic activity and cytotoxicity, does not play a role in kidney retention. C1 NIH,BIOMED ENGN & INSTRUMENTAT PROGRAM,NATL CTR RES RESOURCES,BETHESDA,MD 20892. ALFACELL CORP,BLOOMFIELD,NJ 07003. NINCDS,BIOCHEM SECT,SURG NEUROL BRANCH,NIH,BETHESDA,MD 20892. NR 32 TC 49 Z9 49 U1 0 U2 2 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 0008-5472 J9 CANCER RES JI Cancer Res. PD SEP 15 PY 1996 VL 56 IS 18 BP 4180 EP 4186 PG 7 WC Oncology SC Oncology GA VG370 UT WOS:A1996VG37000020 PM 8797589 ER PT J AU Keane, MM Lowrey, GA Ettenberg, SA Dayton, MA Lipkowitz, S AF Keane, MM Lowrey, GA Ettenberg, SA Dayton, MA Lipkowitz, S TI The protein tyrosine phosphatase DEP-1 is induced during differentiation and inhibits growth of breast cancer cells SO CANCER RESEARCH LA English DT Article ID MOUSE ERYTHROLEUKEMIA-CELLS; LEUKOCYTE-COMMON ANTIGEN; MOLECULAR-CLONING; CHROMOSOMAL LOCALIZATION; SEQUENCE HOMOLOGY; SODIUM-BUTYRATE; CYTOSKELETAL PROTEIN-4.1; EPITHELIAL-CELLS; CARCINOMA-CELLS; PTP-MU AB Sodium butyrate-induced differentiation of breast cancer cell lines was used to identify protein tyrosine phosphatases (PTPs) involved in differentiation and growth inhibition of breast cancer cells. Of 42 PTPs analyzed, 31 were expressed in the ZR75-1 breast cancer cell line, Expression of four PTPs (DEP-1, SAP, PTP gamma, and PAC) was regulated in ZR75-1 cells undergoing differentiation. Expression of two of these PTPs (DEP-1 and SAP) was also regulated in the SKBr-3 cell line undergoing differentiation. In view of its marked induction sith differentiation in an estrogen receptor (ER)-positive and an ER-negative breast cancer cell line, DEP-1 was investigated for a role in growth inhibition or induction of differentiation in breast cancer cells. A DEP-1 cDNA construct under control of a constitutively active cytomegalovirus promoter was transfected into the ZR75-1, SKBR-3, and MCF-7 breast cancer cell lines, and resistant colonies were selected with G418, DEP-1 expression inhibited the development of resistant colonies by 3-5-fold in all three lines compared to transfection with vector alone, Three stable MCF-7 cell lines expressing DEP-1 under control of an inducible metallothionein promoter mere then established, In these lines, induction of DEP-1 expression inhibited breast cancer cell growth by 5-10-fold. These data describe PTPs expressed and regulated in breast cancer cell lines during: differentiation and identify one PTP, DEP-1, that inhibits the growth of breast cancer cells in vitro. C1 NCI,NAVY MED ONCOL BRANCH,NATL NAVAL MED CTR,BETHESDA,MD 20889. LOUISIANA STATE UNIV,MED CTR,CTR EXCELLENCE CANC RES,SHREVEPORT,LA 71130. NR 73 TC 112 Z9 112 U1 1 U2 2 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 0008-5472 J9 CANCER RES JI Cancer Res. PD SEP 15 PY 1996 VL 56 IS 18 BP 4236 EP 4243 PG 8 WC Oncology SC Oncology GA VG370 UT WOS:A1996VG37000029 PM 8797598 ER PT J AU Lin, L Daugherty, B Schlom, J Pestka, S AF Lin, L Daugherty, B Schlom, J Pestka, S TI Construction of phosphorylatable monoclonal antibody to a tumor-associated antigen SO CANCER RESEARCH LA English DT Article ID INTERFERON-ALPHA; EXPRESSION; CELLS; ENHANCEMENT; SPECTRUM; CLONING; TAG-72; INVIVO AB A phosphorylation site was introduced into chimeric monoclonal antibody B72.3 (MAb-chB72.3) by site-specific mutation of the coding sequence, The phosphorylation site for the cAMP-dependent protein kinase was positioned at the carboxyl terminus of the heavy-chain constant region of MAb-chB72.3. The resultant modified MAb-chB72.3-P was expressed in 293 cells and purified, The MAb-chB72.3-P protein was phosphorylated by the catalytic subunit of cAMP-dependent protein kinase with [gamma-P-32]ATP to high radiospecific activity, The P-32-labeled MAb-chB72.3-P protein bound to cells ex-pressing the tumor-associated glycoprotein 72 antigen, The introduction of phosphorylation sites into MAbs provides a new type of MAb for the diagnosis and treatment of cancers. C1 UNIV MED & DENT NEW JERSEY,ROBERT WOOD JOHNSON MED SCH,DEPT MOL GENET & MICROBIOL,PISCATAWAY,NJ 08854. MERCK RES LABS,DEPT INFLAMMAT RES,RAHWAY,NJ 07065. NCI,BETHESDA,MD 20892. FU NCI NIH HHS [R01 CA46465, R01 CA52363]; NIAID NIH HHS [R01 AI36450] NR 24 TC 8 Z9 8 U1 1 U2 2 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 0008-5472 J9 CANCER RES JI Cancer Res. PD SEP 15 PY 1996 VL 56 IS 18 BP 4250 EP 4254 PG 5 WC Oncology SC Oncology GA VG370 UT WOS:A1996VG37000031 PM 8797600 ER PT J AU Cohen, SM Ellwein, LB AF Cohen, SM Ellwein, LB TI Cell proliferation as a major risk factor for cancer: A concept of doubtful validity. SO CANCER RESEARCH LA English DT Letter ID BLADDER-CANCER; CARCINOGENESIS; INDUCTION C1 NEI,NIH,BETHESDA,MD 20892. RP Cohen, SM (reprint author), UNIV NEBRASKA,MED CTR,DEPT PATHOL & MICROBIOL,600 S 42ND ST,OMAHA,NE 68198, USA. NR 10 TC 7 Z9 7 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 0008-5472 J9 CANCER RES JI Cancer Res. PD SEP 15 PY 1996 VL 56 IS 18 BP 4269 EP 4270 PG 2 WC Oncology SC Oncology GA VG370 UT WOS:A1996VG37000036 PM 8797605 ER PT J AU London, SJ Navidi, WC AF London, SJ Navidi, WC TI Lung cancer risk in African-Americans in relation to a race-specific CYP1A1 polymorphism - Reply SO CANCER RESEARCH LA English DT Letter C1 UNIV SO CALIF,SCH MED,DEPT PREVENT MED,LOS ANGELES,CA 90033. RP London, SJ (reprint author), NIEHS,EPIDEMIOL BRANCH,RES TRIANGLE PK,NC 27709, USA. NR 8 TC 1 Z9 1 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 0008-5472 J9 CANCER RES JI Cancer Res. PD SEP 15 PY 1996 VL 56 IS 18 BP 4276 EP 4277 PG 2 WC Oncology SC Oncology GA VG370 UT WOS:A1996VG37000040 ER PT J AU Jeang, KT Rauscher, FJ AF Jeang, KT Rauscher, FJ TI The XVIII Symposium of the International Association for Comparative Research on Leukemia & Related Diseases (IACRLRD): Leukemia and lymphoma pathogenesis and treatment molecular aspects SO CANCER RESEARCH LA English DT Editorial Material C1 WISTAR INST ANAT & BIOL,PHILADELPHIA,PA 19104. NIAID,MOL VIROL SECT,MOL MICROBIOL LAB,NIH,BETHESDA,MD 20892. RI Jeang, Kuan-Teh/A-2424-2008 NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI PHILADELPHIA PA PUBLIC LEDGER BLDG, SUITE 816, 150 S. INDEPENDENCE MALL W., PHILADELPHIA, PA 19106 SN 0008-5472 J9 CANCER RES JI Cancer Res. PD SEP 15 PY 1996 VL 56 IS 18 BP 4288 EP 4292 PG 5 WC Oncology SC Oncology GA VG370 UT WOS:A1996VG37000043 PM 8797610 ER PT J AU Carter, CRD Sayers, TJ Wiltrout, RH TurcovskiCorrales, SM Taub, DD AF Carter, CRD Sayers, TJ Wiltrout, RH TurcovskiCorrales, SM Taub, DD TI Generation of antigenic peptides by lymphocyte granule serine proteases (granzymes) SO CELLULAR IMMUNOLOGY LA English DT Article ID CLASS-I MOLECULES; CYTOTOXIC-T-CELLS; HISTOCOMPATIBILITY COMPLEX-MOLECULES; FREE SYNTHETIC PEPTIDE; NATURAL-KILLER-CELLS; PORE-FORMING PROTEIN; INFLUENZA NUCLEOPROTEIN; DNA FRAGMENTATION; MHC MOLECULES; IMMUNOGENIC PEPTIDES AB We have examined the ability of several serine proteases (granzymes) isolated from the granules of the rat natural killer cell line, RNK, to generate antigenic peptides of ovalbumin (Oval that are capable of being recognized by Ova-specific CD8(+) T cells. The mouse MHC class I-restricted cytotoxic T-cell clone, GX-1, which recognizes a trypsinized fragment of Ova in the context of H-2(b), was able to lyse EL4 (H-2(b)) target cells in the presence of Ova and the granzymes but not in the presence of Ova or granzymes alone. Similar results were obtained using the murine Ova-specific CD8(+) T cell hybridomas, RF33.70 and CD80VA. In all cases, the T cells' responses were MHC class I-restricted as Ova:granzyme mixtures failed to mediate the lysis of the MHC-disparate target cell, P815 (H-2(d)), The purified rat granzyme preparations contained three distinct enzymatic specificities: asp-ase, met;ase, and tryptase. Aprotinin, a protease inhibitor that only inhibits tryptase activity in vitro, completely abolished the CD8(+) T-cell responses to Ova. These results, along with peptide loading studies using the RMA-S cell line, suggest that the granzyme treatment of Ova can generate the proper antigenic fragments which facilitate class I-restricted CTL responses both in vivo and in vitro. We believe that enzymes produced and released by NK or cytotoxic T cells within a tissue microenvironment may enhance the cleavage of-target cell antigens as well as soluble antigens resulting in the improved uptake and processing of soluble antigens. (C) 1996 Academic Press, Inc. C1 NCI,BCDP,FREDERICK CANC RES & DEV CTR,FREDERICK,MD 21702. SAIC FREDERICK,EXPT IMMUNOL LAB,DIV BASIC SCI,FREDERICK,MD 21702. SAIC FREDERICK,BIOL CARCINOGENESIS & DEV PROGRAM,FREDERICK,MD 21702. SAIC FREDERICK,CLIN SERV PROGRAM,FREDERICK,MD 21702. RI Sayers, Thomas/G-4859-2015 NR 69 TC 8 Z9 8 U1 0 U2 0 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0008-8749 J9 CELL IMMUNOL JI Cell. Immunol. PD SEP 15 PY 1996 VL 172 IS 2 BP 235 EP 245 DI 10.1006/cimm.1996.0238 PG 11 WC Cell Biology; Immunology SC Cell Biology; Immunology GA VJ542 UT WOS:A1996VJ54200013 PM 8964086 ER PT J AU Fananapazir, L Packer, D Prystowsky, EN AF Fananapazir, L Packer, D Prystowsky, EN TI Differential effects of changes in local myocardial refractoriness on atrial and ventricular latency SO CIRCULATION LA English DT Article DE atrium; conduction; electrophysiology; myocardium; ventricles ID ACTION-POTENTIAL DURATION; TRANSIENT OUTWARD CURRENT; PARKINSON-WHITE SYNDROME; CANINE CARDIAC PURKINJE; CYCLE LENGTH; CONDUCTION-VELOCITY; GUINEA-PIG; HEART-RATE; CELLS; REPOLARIZATION AB Background Assessment of myocardial refractoriness and conduction properties. critical to development and propagation of reentrant arrhythmias, is an integral part of the investigation of atrial and ventricular tachycardias through the use of programmed electrical stimulation. Local conduction itself, however, may be affected by myocardial refractoriness. Methods and Results We studied the effects of changes in myocardial refractoriness on local conduction in right atrial and ventricular myocardium in 19 patients. Changes in effective, functional, and relative refractoriness were accomplished with the use of four pacing protocols, including drive train pacing cycle lengths (PCLs) of 600 and 400 milliseconds (ms) and drive train durations (DTDs) of 8 and 50 stimuli. Unipolar cathodal stimulation was performed from the distal electrode, and unipolar electrograms were recorded from the proximal three poles of quadripolar catheters with 5-mm interelectrode spaces. Atrial and ventricular effective and relative refractory periods (ERPs and RRPs) were significantly shortened by bulb the reduction in PCL and the increase in DTD. The reduction in the PCL shortened atrial and ventricular refractory periods significantly more than did the increase in the DTD. Changes in ventricular refractory periods were significantly greater compared with atrial refractory periods. The ratio or RRP to ERP was reduced in the atrium but significantly increased in the ventricle with reduction in PCL and increase in DTD. For all premature intervals, the conduction interval from stimulus to the first recording electrode pole was significantly greater than conduction intervals measured between subsequent electrode poles. The greatest increase in conduction interval with closely coupled premature complexes occurred between the stimulus artifact and the first recording electrode pole. Reduction in ventricular but not atrial ERP was associated with significantly increased local conduction interval. Conclusions First, most of the conduction delay after the stimulus artifact occurs within 5 mm from the pacing site. Second, closely coupled premature complexes delay conduction primarily by prolonging latency in the first 5 mm from the pacing site. Third, fundamental differences occur between the atrium and ventricle regarding changes in local conduction as a function of changes in ERP, suggesting that factors involved in sudden changes in refractoriness (eg, heart rate acceleration) could produce divergent effects on atrial and ventricular arrhythmogenesis. C1 NHLBI,CLIN ELECTROPHYSIOL LAB,CARDIOL BRANCH,NIH,BETHESDA,MD 20892. DUKE UNIV,MED CTR,DEPT MED,DIV CARDIOL,DURHAM,NC 27710. MAYO CLIN & MAYO FDN,ROCHESTER,MN 55905. NORTHSIDE CARDIOL,INDIANAPOLIS,IN. RP Fananapazir, L (reprint author), NHLBI,INHERITED CARDIAC DIS SECT,CARDIOL BRANCH,NIH,BLDG 10,RM 7B-14,10 CTR DR,MSC 1650,BETHESDA,MD 20892, USA. NR 50 TC 3 Z9 3 U1 0 U2 1 PU AMER HEART ASSOC PI DALLAS PA 7272 GREENVILLE AVENUE, DALLAS, TX 75231-4596 SN 0009-7322 J9 CIRCULATION JI Circulation PD SEP 15 PY 1996 VL 94 IS 6 BP 1364 EP 1371 PG 8 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA VG350 UT WOS:A1996VG35000034 PM 8822994 ER PT J AU KimSchulze, S McGowan, KA Hubchak, SC Cid, MC Martin, MB Kleinman, HK Greene, GL Schnaper, HW AF KimSchulze, S McGowan, KA Hubchak, SC Cid, MC Martin, MB Kleinman, HK Greene, GL Schnaper, HW TI Expression of an estrogen receptor by human coronary artery and umbilical vein endothelial cells SO CIRCULATION LA English DT Article DE estrogen; endothelium; receptor; cells ID BREAST-CANCER CELLS; VASCULAR SMOOTH-MUSCLE; STEROID-HORMONES; ESTRADIOL; BINDING; FOS; ANTIESTROGENS; PROGESTERONE; INHIBITION; HOMODIMER AB Background Premenopausal women have much lower susceptibility to coronary artery disease than do men or postmenopausal women. It has been proposed that estrogen plays a role in cardioprotection, but little information is available regarding the mechanism by which estrogen may help to protect the vasculature. Here, we describe an estrogen receptor (ER) in human coronary artery and umbilical vein endothelial cells. Methods and Results Human umbilical vein endothelial cells and human coronary artery endothelial cells were cultured in hormone-free medium for 48 hours before experiments. Estradiol (3.7 nmol/L) added to cultures promoted proliferation by a mechanism that is inhibited by the specific ER antagonist ICI182,780. Estradiol-treated cells incorporated twice the [H-3]thymidine of hormone-free cells; this increase was prevented by ICI182,780. Endothelial cells from both sources stained in a nuclear pattern with an ER-specific antibody. Ribonuclease protection assay detected mRNA for the ER. Ligand-binding studies estimated 2x10(4) to 8x10(4) receptors per cell and a K-d of approximate to 5 nmol/L. Interaction of ERs with a consensus estrogen response element was shown by an electrophoretic mobility shift assay,.In addition, an antibody against the ER supershifted the protein-DNA complex. Conclusions These studies define the presence of an ER in human coronary artery and umbilical vein endothelial cells. They support the hypothesis that cardioprotective effects of estrogen are mediated, at least in part. through a classic steroid hormone receptor mechanism. C1 NORTHWESTERN UNIV,SCH MED,DEPT PEDIAT,CHICAGO,IL 60611. NIDR,DEV BIOL LAB,NIH,BETHESDA,MD 20892. FDN CLIN,DEPT INTERNAL MED,BARCELONA,SPAIN. GEORGETOWN UNIV,SCH MED,LOMBARDI CANC CTR,WASHINGTON,DC. UNIV CHICAGO,BEN MAY INST,CHICAGO,IL 60637. OI Cid Xutgla, Maria Cinta/0000-0002-4730-0938 FU NHLBI NIH HHS [HL-53918, R01 HL053918]; NIAMS NIH HHS [AR-30692]; NIDDK NIH HHS [DK-49362] NR 38 TC 158 Z9 161 U1 0 U2 0 PU AMER HEART ASSOC PI DALLAS PA 7272 GREENVILLE AVENUE, DALLAS, TX 75231-4596 SN 0009-7322 J9 CIRCULATION JI Circulation PD SEP 15 PY 1996 VL 94 IS 6 BP 1402 EP 1407 PG 6 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA VG350 UT WOS:A1996VG35000039 PM 8822999 ER PT J AU Nicholson, J Kidd, P Mandy, F Livnat, D Kagan, J AF Nicholson, J Kidd, P Mandy, F Livnat, D Kagan, J TI Three-color supplement to the NIAID DAIDS guideline for flow cytometric immunophenotyping SO CYTOMETRY LA English DT Article ID SUBSETS; AIDS AB Since the publication of the ''NIAID DAIDS Guideline for Flow Cytometric Immunophenotyping'' (1) in 1993, significant scientific and technological advances in the development and production of reagents, instrumentation, and software have permitted widespread access to multicolor flow cytometry in both research and clinical laboratories. As is often the case with complex new technologies, each advance, while opening up exciting new capabilities, can also bring new sources of variability and a commensurate increase in the requirements for quality assurance at all levels. The purpose of this document is to update the 1993 NIAID DAIDS Guideline with new recommendations designed to help standardize the methodology and minimize measurement variability in determining patients' CD4 and CD8 T-cell counts using 3-color flow cytometry. Issues pertaining to specimen collection and handling, problematic specimens, hematology, and laboratory performance assessment were addressed in the previous guideline.(1) In assembling this supplement, factors such as reduced technician time, decreased need for isotype controls, fewer assay tubes, and the potential for diminished cost were considered advantages of 3-color (vs. 2-color) flow cytometry. In contrast, the increased complexity of factors such as spectral compensation, instrument set up, and data collection/analysis were considered disadvantages. In addition, some antibody combinations are not commercially available, and certain third-color fluorochromes may not be optimal with all flow cytometers. The specifications and recommendations contained herein were developed for use in laboratories that support clinical trials and epidemiologic studies done under the auspices of the National Institute of Allergy and Infectious Diseases, Division of AIDS (NIAID DAIDS). Efforts currently underway by the Centers for Disease Control and Prevention and by the National Committee for Clinical Laboratory Standards (NCCLS) will likely provide additional guidance in this rapidly changing arena. C1 NIAID,DIV AIDS,NIH,ROCKVILLE,MD 20852. CTR DIS CONTROL & PREVENT,ATLANTA,GA 30341. UNIV MED & DENT NEW JERSEY,ROBERT WOOD JOHNSON MED SCH,NEW BRUNSWICK,NJ. HLTH & WELF CANADA,OTTAWA,ON,CANADA. FU NIAID NIH HHS [N01-AI-45175] NR 5 TC 58 Z9 60 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC 605 THIRD AVE, NEW YORK, NY 10158-0012 SN 0196-4763 J9 CYTOMETRY JI Cytometry PD SEP 15 PY 1996 VL 26 IS 3 BP 227 EP 230 DI 10.1002/(SICI)1097-0320(19960915)26:3<227::AID-CYTO8>3.0.CO;2-B PG 4 WC Biochemical Research Methods; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA VH823 UT WOS:A1996VH82300008 PM 8889396 ER PT J AU Shao, RG Shimizu, T Pommier, Y AF Shao, RG Shimizu, T Pommier, Y TI Brefeldin A is a potent inducer of apoptosis in human cancer cells independently of p53 SO EXPERIMENTAL CELL RESEARCH LA English DT Article ID TOPOISOMERASE-II INHIBITORS; ADP-RIBOSYLATION FACTOR; LEUKEMIC HL-60 CELLS; GTP-BINDING PROTEIN; DNA FRAGMENTATION; RAT HEPATOCYTES; INTRACELLULAR-TRANSPORT; DIFFERENTIAL INDUCTION; SPHINGOMYELIN PATHWAY; GOLGI MEMBRANES AB Brefeldin A (BFA) is a natural product that affects the structure and function of the Golgi apparatus and is in development for cancer chemotherapy. We observed that a wide range of cancer cells could undergo DNA fragmentation associated with apoptosis after BFA treatment. This DNA fragmentation was induced within 15 h in HL60 leukemia cells and after 48 h in K562 leukemia and HT-29 colon carcinoma cells with BFA concentrations as low as 0.1 mu M. The DNA fragmentation had the typical internucleosomal pattern in HL60 and HT-29 cells. Apoptotic cells were also detected by microscopy. BFA-induced apoptosis is p53-independent as HL60 and K562 cells are p53 null and HT-29 are p53 mutant cells. BFA could potentiate UCN-01 and staurosporine-induced DNA fragmentation in HL60 cells. Cyclin B1/Cdc2 kinase activity decreased after BFA treatment in HL60 cells, indicating that BFA-induced DNA fragmentation was independent of a cyclin B1/Cdc2 kinase upregulation pathway. Cycloheximide could not prevent BFA-induced DNA fragmentation in HL60 cells, suggesting that protein synthesis is not needed for HL60 cells to undergo apoptosis. On the contrary, cycloheximide blocked BFA-induced DNA fragmentation in HT-29 cells, indicating that apoptosis in HT-29 cells requires macromolecular synthesis. Cell-free system experiments suggested that cytosolic proteins play an important role in triggering DNA fragmentation during apoptosis induced by BFA. Our results show that transduction signaling pathways play central roles in apoptotic regulation. (C) 1996 Academic Press, Inc. C1 NCI, DNA TOPOL SECT,MOL PHARMACOL LAB, DEV THERAPEUT PROGRAM,DIV CANC TREATMENT, BETHESDA, MD 20892 USA. NR 46 TC 85 Z9 86 U1 2 U2 5 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0014-4827 J9 EXP CELL RES JI Exp. Cell Res. PD SEP 15 PY 1996 VL 227 IS 2 BP 190 EP 196 DI 10.1006/excr.1996.0266 PG 7 WC Oncology; Cell Biology SC Oncology; Cell Biology GA VK395 UT WOS:A1996VK39500003 PM 8831555 ER PT J AU Movsesyan, V Whalin, M Shibutani, M Katagiri, Y Broude, E Guroff, G AF Movsesyan, V Whalin, M Shibutani, M Katagiri, Y Broude, E Guroff, G TI Down-regulation of cyclin F levels during nerve growth factor-induced differentiation of PC12 cells SO EXPERIMENTAL CELL RESEARCH LA English DT Article ID PHEOCHROMOCYTOMA CELLS; NEURONAL DIFFERENTIATION; PHOSPHORYLATION; EXPRESSION; INDUCTION; KINASES; K-252A; LINE AB Treatment with nerve growth factor (NGF) produces a marked decrease of cyclin F levels in PC12EY cells. This decrease is prevented by the simultaneous addition of K-252a. A smaller decrease is observed when the cells are treated with fibroblast growth factor, but the addition of epidermal growth factor has no comparable effect; Time course studies show that the decrease in cyclin F precedes the changes produced by NGF in the distribution of cells within the cell cycle. The data suggest that cyclin F is involved in NGF-mediated cell cycle events during the differentiation of PC12EY cells. (C) 1996 Academic Press, Inc. C1 NICHHD,GROWTH FACTORS SECT,NIH,BETHESDA,MD 20892. RI Shibutani, Makoto/C-2510-2013 NR 27 TC 6 Z9 6 U1 0 U2 0 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0014-4827 J9 EXP CELL RES JI Exp. Cell Res. PD SEP 15 PY 1996 VL 227 IS 2 BP 203 EP 207 DI 10.1006/excr.1996.0268 PG 5 WC Oncology; Cell Biology SC Oncology; Cell Biology GA VK395 UT WOS:A1996VK39500005 PM 8831557 ER PT J AU Hasson, T Skowron, JF Gilbert, DJ Avraham, KB Perry, WL Bement, WM Anderson, BL Sherr, EH Chen, ZY Greene, LA Ward, DC Corey, DP Mooseker, MS Copeland, NG Jenkins, NA AF Hasson, T Skowron, JF Gilbert, DJ Avraham, KB Perry, WL Bement, WM Anderson, BL Sherr, EH Chen, ZY Greene, LA Ward, DC Corey, DP Mooseker, MS Copeland, NG Jenkins, NA TI Mapping of unconventional myosins in mouse and human SO GENOMICS LA English DT Article ID GENETIC-LINKAGE MAP; AMYOTROPHIC-LATERAL-SCLEROSIS; INSITU HYBRIDIZATION; HEAVY-CHAIN; CHROMOSOMAL LOCALIZATION; PARTIAL ALBINISM; MESSENGER-RNA; DNA-SEQUENCES; PROTEIN; CLONING AB Myosins are molecular motors that move along filamentous actin. Seven classes of myosin are expressed in vertebrates: conventional myosin, or myosin-II, as well as the 6 unconventional myosin classes -I, -V, -VI, VII, -IX, and -X. We have mapped in mouse 22 probes encompassing all known unconventional myosins and, as a result, have identified 16 potential unconventional myosin genes. These genes include 7 myosins-I, 2 myosins-V, 1 myosin-VI, 3 myosins-VII, 2 myosins-IX, and 1 myosin-X. The map location of 5 of these genes was identified in human chromosomes by fluorescence in situ hybridization. (C) 1996 Academic Press, Inc. C1 NCI,MAMMALIAN GENET LAB,ABL BASIC RES PROGRAM,FREDERICK CANC RES & DEV CTR,FREDERICK,MD 21702. YALE UNIV,DEPT BIOL,NEW HAVEN,CT 06520. YALE UNIV,DEPT PATHOL,NEW HAVEN,CT 06520. YALE UNIV,DEPT CELL BIOL,NEW HAVEN,CT 06520. YALE UNIV,DEPT GENET,NEW HAVEN,CT 06520. UNIV WISCONSIN,DEPT ZOOL,MADISON,WI 53706. COLUMBIA UNIV,DEPT PATHOL,NEW YORK,NY 10032. MASSACHUSETTS GEN HOSP,HOWARD HUGHES MED INST,BOSTON,MA 02114. OI Corey, David/0000-0003-4497-6016 FU NIDDK NIH HHS [DK38979, DK25387]; NINDS NIH HHS [NS33689] NR 63 TC 66 Z9 66 U1 0 U2 4 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0888-7543 J9 GENOMICS JI Genomics PD SEP 15 PY 1996 VL 36 IS 3 BP 431 EP 439 DI 10.1006/geno.1996.0488 PG 9 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA VH856 UT WOS:A1996VH85600007 PM 8884266 ER PT J AU Ropp, PA Copeland, WC AF Ropp, PA Copeland, WC TI Cloning and characterization of the human mitochondrial DNA polymerase, DNA polymerase gamma SO GENOMICS LA English DT Article ID MISMATCH REPAIR; CELL-LINES; MICROSATELLITE INSTABILITY; ESCHERICHIA-COLI; CAG REPEAT; NUDE MICE; GENE; ESTABLISHMENT; PURIFICATION; REPLICATION AB The nuclear-encoded DNA polymerase gamma (DNA POL gamma) is the sole DNA polymerase required for the replication of the mitochondrial DNA. We have cloned the cDNA for human DNA POL gamma and have mapped the gene to the chromosomal location 15q24. Additionally, the DNA POL gamma gene from Drosophila melanogaster and a partial cDNA for DNA POL gamma from Gallus gallus have been cloned. The predicted human DNA POL gamma polypeptide is 1239 amino acids, with a calculated molecular mass of 139.5 kDa. The human amino acid sequence is 41.6, 43.0, 48.7, and 77.6% identical to those of Schizosaccharomyces pombe, Saccharomyces cerevisiae, Drosophila melanogaster, and the C-terminal half of G. gallus, respectively. Polyclonal antibodies raised against the polymerase portion of the protein reacted specifically with a 140-kDa protein in mitochondrial extracts and immunoprecipitated a protein with DNA POL gamma like activity from mitochondrial extracts. The human DNA POL gamma is unique in that the first exon of the gene contains a CAG(10) trinucleotide repeat. (C) 1996 Academic Press, Inc. C1 NIEHS,MOL GENET LAB,RES TRIANGLE PK,NC 27709. NR 51 TC 187 Z9 196 U1 0 U2 4 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0888-7543 J9 GENOMICS JI Genomics PD SEP 15 PY 1996 VL 36 IS 3 BP 449 EP 458 DI 10.1006/geno.1996.0490 PG 10 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA VH856 UT WOS:A1996VH85600009 PM 8884268 ER PT J AU Miki, T Taira, M Hockman, S Shimada, F Lieman, J Napolitano, M Ward, D Taira, M Makino, H Manganiello, VC AF Miki, T Taira, M Hockman, S Shimada, F Lieman, J Napolitano, M Ward, D Taira, M Makino, H Manganiello, VC TI Characterization of the cDNA and gene encoding human PDE3B, the cGIP1 isoform of the human cyclic GMP-inhibited cyclic nucleotide phosphodiesterase family SO GENOMICS LA English DT Article ID BETA-SUBUNIT GENE; CAMP-PHOSPHODIESTERASE; DROSOPHILA-MELANOGASTER; AMP PHOSPHODIESTERASE; MOLECULAR-CLONING; NONSENSE MUTATION; INSULIN; PURIFICATION; DNA; PHOSPHORYLATION AB Two distinct PDE3 [cyclic GMP-inhibited cyclic nucleotide phosphodiesterase (cGI PDE)] isoforms, cGIP1 and cGIP2, have been identified. Here we report cloning of the cDNA and gene encoding human (H)cGIP1 (classified as PDE3B). The cDNA encodes a protein of 1112 amino acids (similar to 123 kDa). Northern blots indicate that its mRNA is expressed in several adipose tissue depots. The human PDE3B gene is composed of 16 exons spanning more than 114 kb and was localized to chromosome 11p15 by in situ hybridization. Exon/intron boundaries were determined, and genetic polymorphism, confirmed by single-strand conformational polymorphism of DNA from 25 healthy subjects, was demonstrated in exon 4 at nucleotide 1389 (A/G). Two polymorphic dinucleotide repeat sequences were identified in introns 5 and 12. (C) 1996 Academic Press, Inc. C1 CHIBA UNIV,SCH MED,DEPT INTERNAL MED 2,CHUO KU,CHIBA 260,JAPAN. NICHHD,MOL GENET LAB,BETHESDA,MD 20892. NHLBI,PULM CRIT CARE MED BRANCH,BETHESDA,MD 20892. YALE UNIV,DEPT GENET,NEW HAVEN,CT 06510. NR 41 TC 47 Z9 50 U1 0 U2 1 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0888-7543 J9 GENOMICS JI Genomics PD SEP 15 PY 1996 VL 36 IS 3 BP 476 EP 485 DI 10.1006/geno.1996.0493 PG 10 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA VH856 UT WOS:A1996VH85600012 PM 8884271 ER PT J AU Stefanova, I Saville, MW Peters, C Cleghorn, FR Schwartz, D Venzon, DJ Weinhold, KJ Jack, N Bartholomew, C Blattner, WA Yarchoan, R Bolen, JB Horak, ID AF Stefanova, I Saville, MW Peters, C Cleghorn, FR Schwartz, D Venzon, DJ Weinhold, KJ Jack, N Bartholomew, C Blattner, WA Yarchoan, R Bolen, JB Horak, ID TI HIV infection-induced posttranslational modification of T cell signaling molecules associated with disease progression SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article DE HIV; T cell receptor; tyrosine phosphorylation; signal transduction; protein tyrosine kinases ID HUMAN-IMMUNODEFICIENCY-VIRUS; TYROSINE PROTEIN-KINASES; LYMPHOCYTES-T; HIV-1-INFECTED PATIENTS; ANTIGEN RECEPTOR; MICE LACKING; GLUTATHIONE; ACTIVATION; PHOSPHORYLATION; INDIVIDUALS AB In attempt to elucidate the mechanism of the HIV infection induced T cell unresponsiveness, we studied signal-transducing molecules proximal to the T cell receptor (TCR) in T lymphocytes of HIV-infected individuals, Total amounts of protein tyrosine kinases (PTKs) Lck, Fyn, and ZAP-70 and the zeta chain of the TCR were found significantly decreased in T cells of symptomatic/AIDS patients as well as in T cells of individuals in acute and early asymptomatic stages of HIV infection, Unexpectedly, the detection of Lck, Fyn, and ZAP-70 was reversed after the treatment of cell lysates with dithiothreitol, This suggests that PTKs Lck, Fyn, and ZAP-70 were modified by a mechanism altering the status of sulfhydryl groups, Moreover, this mechanism seems to affect selectively T cells of HIV infected patients since B cell PTKs Syk and Lyn were detected structurally and functionally intact, Interestingly, similar alterations of signaling molecules were not detected in T cells of HIV-infected long-term asymptomatic individuals, Modification of T cell PTKs may thus underlie the HIV-induced impairment of lymphocyte function and may potentially predict disease progression. C1 NCI,METAB BRANCH,NIH,BETHESDA,MD 20892. NCI,MED BRANCH,NIH,BETHESDA,MD 20892. NCI,VIRAL EPIDEMIOL SECT,NIH,BETHESDA,MD 20892. NCI,BIOSTAT & DATA MANAGEMENT SECT,NIH,BETHESDA,MD 20892. JOHNS HOPKINS UNIV,SCH HYG & PUBL HLTH,DEPT MOL MICROBIOL & IMMUNOL,BALTIMORE,MD 21205. DUKE UNIV,MED CTR,DEPT SURG,DURHAM,NC 27710. UNIV W INDIES,ST AUGUSTINE,TRINID & TOBAGO. DNAX RES INST MOL & CELLULAR BIOL INC,PALO ALTO,CA 94304. RI Venzon, David/B-3078-2008 NR 56 TC 76 Z9 76 U1 0 U2 3 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 222 E 70TH STREET, NEW YORK, NY 10021 SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD SEP 15 PY 1996 VL 98 IS 6 BP 1290 EP 1297 DI 10.1172/JCI118915 PG 8 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA VJ484 UT WOS:A1996VJ48400005 PM 8823293 ER PT J AU Lefkowith, JB Kiehl, M Rubenstein, J DiValerio, R Bernstein, K Kahl, L Rubin, RL Gourley, M AF Lefkowith, JB Kiehl, M Rubenstein, J DiValerio, R Bernstein, K Kahl, L Rubin, RL Gourley, M TI Heterogeneity and clinical significance of glomerular-binding antibodies in systemic lupus erythematosus SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article DE lupus nephritis; autoantibodies; glomerular basement membrane; systemic lupus erythematosus; nucleosomes ID ANTI-DNA ANTIBODIES; NUCLEOSOME-RESTRICTED ANTIBODIES; FORM IMMUNE DEPOSITS; TISSUE-BASED ELISA; MRL-LPR MICE; BASEMENT-MEMBRANE; RENAL-DISEASE; MONOCLONAL-ANTIBODIES; CELL-TYPES; NEPHRITIS AB We used an ELISA employing extracts of human glomerular basement membrane (GEM) to detect, characterize, and evaluate the clinical significance of glomerular-binding IgG in patients with SLE nephritis. Most patients with SLE nephritis exhibited GEM-binding IgG, although many patients with active nonrenal SLE or symptomatic, drug-induced lupus had similar reactivity, albeit at lower levels. IgG binding to GEM in SLE nephritis patients was decreased by DNase pretreatment of GEM, restored after DNase with nuclear antigens (most notably with nucleosomes), inhibited by exogenous nuclear antigens (particularly nucleosomes), but unaffected by exposure of serum to DNase/high ionic strength. The characteristics of IgG binding to GEM largely paralleled the patients' underlying autoimmune response, which was dominated either by antibodies to DNA/nucleosomes or to nucleosomes alone. Binding of lupus sera to nonrenal extracellular matrix (even with nucleosomes) was not equivalent to GBM. Collagenase pretreatment of GEM variably decreased IgG binding, depending on the level and type of binding. SLE nephritis patients with high levels of GEM-binding IgG exhibited more severe disease clinically, but the same renal histopathology, as patients with lower levels. The level of GEM-binding IgG at presentation did not predict the therapeutic response, but decreased in responders to therapy. In sum, glomerular-binding IgG in lupus nephritis binds to epitopes on chromatin, which adheres to GEM in part via collagen. These autoantibodies appear necessary, but not sufficient, for the development of nephritis, and correlate with clinical rather than histopathologic parameters of disease activity. C1 WASHINGTON UNIV, SCH MED, DEPT MOL BIOL & PHARMACOL, ST LOUIS, MO 63110 USA. NIAMSD, NIH, BETHESDA, MD 20892 USA. Scripps Res Inst, DEPT MOL & EXPT MED, LA JOLLA, CA 92037 USA. RP Lefkowith, JB (reprint author), WASHINGTON UNIV, SCH MED, DIV RHEUMATOL, DEPT MED, BOX 8045, ST LOUIS, MO 63110 USA. NR 48 TC 65 Z9 66 U1 0 U2 1 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 222 E 70TH STREET, NEW YORK, NY 10021 SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD SEP 15 PY 1996 VL 98 IS 6 BP 1373 EP 1380 DI 10.1172/JCI118924 PG 8 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA VJ484 UT WOS:A1996VJ48400014 PM 8823302 ER PT J AU Heiss, JD Papavassiliou, E Merrill, MJ Nieman, L Knightly, JJ Walbridge, S Edwards, NA Oldfield, EH AF Heiss, JD Papavassiliou, E Merrill, MJ Nieman, L Knightly, JJ Walbridge, S Edwards, NA Oldfield, EH TI Mechanism of dexamethasone suppression of brain tumor-associated vascular permeability in rats - Involvement of the glucocorticoid receptor and vascular permeability factor SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article DE cerebral edema; vascular endothelial growth factor; glioma; mifepristone; steroids ID ENDOTHELIAL GROWTH-FACTOR; MESSENGER-RNA STABILITY; BLOOD-BRAIN; MALIGNANT GLIOMA; PROTEIN PRODUCT; FACTOR GENE; CELLS; EXPRESSION; MODEL; ANGIOGENESIS AB Brain tumor-associated cerebral edema arises because tumor capillaries lack normal blood-brain barrier function; vascular permeability factor (VPF, also known as vascular endothelial growth factor, VEGF) is a likely mediator of this phenomenon, Clinically, dexamethasone reduces brain tumor-associated vascular permeability through poorly understood mechanisms. Our goals were to determine if suppression of permeability by dexamethasone might involve inhibition of VPF action or expression, and if dexamethasone effects in this setting are mediated by the glucocorticoid receptor (GR). In two rat models of permeability (peripheral vascular permeability induced by intradermal injection of 9L glioma cell-conditioned medium or purified VPF, and intracerebral vascular permeability induced by implanted 9L glioma), dexamethasone suppressed permeability in a dose-dependent manner. Since 80% of the permeability-inducing activity in 9L-conditioned medium was removed by anti-VPF antibodies, we examined dexamethasone effects of VPF expression in 9L cells. Dexamethasone inhibited FCS- and PDGF-dependent induction of VPF expression. At all levels (intradermal, intracranial, and cell culture), dexamethasone effects were reversed by the GR antagonist mifepristone (RU486). Dexamethasone may decrease brain tumor-associated vascular permeability by two GR-dependent mechanisms: reduction of the response of the vasculature to tumor-derived permeability factors (including VPF), and reduction of VPF expression by tumor cells. C1 NINCDS,SURG NEUROL BRANCH,NIH,BETHESDA,MD 20892. NICHHD,DEV ENDOCRINOL BRANCH,NIH,BETHESDA,MD 20892. NR 70 TC 169 Z9 176 U1 0 U2 3 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 222 E 70TH STREET, NEW YORK, NY 10021 SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD SEP 15 PY 1996 VL 98 IS 6 BP 1400 EP 1408 DI 10.1172/JCI118927 PG 9 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA VJ484 UT WOS:A1996VJ48400017 PM 8823305 ER PT J AU Khattri, R Sperling, AI Qian, DP Fitch, FW Shores, EW Love, PE Bluestone, JA AF Khattri, R Sperling, AI Qian, DP Fitch, FW Shores, EW Love, PE Bluestone, JA TI TCR-gamma delta cells in CD3 zeta-deficient mice contain Fc epsilon RI gamma in the receptor complex but are specifically unresponsive to antigen SO JOURNAL OF IMMUNOLOGY LA English DT Article ID PROTEIN-TYROSINE KINASE; ZETA-CHAIN; LYMPHOCYTES-T; INTRAEPITHELIAL LYMPHOCYTES; SIGNAL-TRANSDUCTION; MONOCLONAL-ANTIBODY; ACTIVATION; IDENTIFICATION; AFFINITY; CD3-ZETA AB Unlike TCR-alpha beta cells, TCR-gamma delta cells express a distinct member of the zeta family, the gamma-chain of Fc epsilon RI (Fc epsilon RI gamma) within the TCR complex. To study the role of the Fc epsilon RI gamma-chain in TCR-gamma delta cells, a TCR-gamma delta transgenic mouse (G8) has been crossed with CD3 zeta-chain-deficient mice (G8.zeta(-/-)). Thy-1(+) spleen and lymph node cells of these animals expressed low levels of CD3/TCR, These results suggested that the zeta-chain is required for effective TCR transport to the cell surface. In contrast, intraepithelial TCR-gamma delta cells of G8.zeta(-/-) mice expressed high levels of TCR. Immunoprecipitation with anti-CDS showed that Fc epsilon RI gamma-chains were associated with the TCR complex in T cells isolated from zeta-deficient mice, Although the Fc epsilon RI gamma-expressing T cells proliferated in response to stimulation by TCR-specific Abs including anti-CD3 epsilon, anti-pan gamma delta, and anti-V gamma 2 mAb, the G8.zeta(-/-) T cells did not respond to the G8-specific Ag(T10(b)), anti-Thy-1 mAb, or Con A, The unresponsiveness to the Ag was not due to the reduced TCR expression, because intraepithelial TCR-gamma delta cells from the zeta-deficient mice did not respond to Ag. The inability of the G8.zeta(-/-) T cells to respond to Ag could not be overcome by providing an anti-CD28 costimulatory signal or by adding exogenous rIL-2, Taken together, our data suggest that the Fc epsilon RI gamma-chain associates with the TCR-gamma delta complex in the absence of the zeta-chain, but it is not able to substitute for the zeta-chain for effective transport of TCR to the cell surface or functional responses to Ag. C1 UNIV CHICAGO,BEN MAY INST CANC RES,DEPT PATHOL,COMM IMMUNOL,CHICAGO,IL 60637. US FDA,CTR BIOL EVALUAT & RES,DIV HEMATOL PROD,BETHESDA,MD 20892. NICHHD,LAB MAMMALIAN GENES & DEV,NIH,BETHESDA,MD 20892. FU NHLBI NIH HHS [T32 HL 07605-11]; NIAID NIH HHS [R01 AI26847] NR 41 TC 8 Z9 11 U1 0 U2 0 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD SEP 15 PY 1996 VL 157 IS 6 BP 2320 EP 2327 PG 8 WC Immunology SC Immunology GA VL103 UT WOS:A1996VL10300013 PM 8805629 ER PT J AU KitagawaSakakida, S Schwartz, RH AF KitagawaSakakida, S Schwartz, RH TI Multifactor cis-dominant negative regulation of IL-2 gene expression in anergized T cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID PROTEIN-TYROSINE PHOSPHORYLATION; CLONAL ANERGY; INTERLEUKIN-2 TRANSCRIPTION; MONOCLONAL-ANTIBODY; LYMPHOCYTES-T; IN-VIVO; ANTIGEN; INDUCTION; AP-1; STIMULATION AB The molecular mechanism underlying IL-2 transcriptional blockade in anergic T cell clones is not fully understood, To examine whether an active negative regulatory process occurs, we created a reporter construct containing as an enhancer four copies of the NF-AT site and one copy of the octamer site (4X NF-AT-Oct), This construct was only slightly reduced (1,3-fold) in its expression when stimulated under anergic conditions, while a whole mouse IL-2 enhancer construct showed a reduction of 4,3-fold, Addition of the -176 to -96 sequence to the 4X NF-AT-Oct construct did not impart the ability to be affected by anergy, but addition of the -236 to -96 sequence did, demonstrating that anergy is an active inhibitory process and that more than the presence of the -150 AP-1 binding site (-52 to -147) is required to mediate the effect. Mutational studies of the -236 to -96 sequence indicated that the presence of both the -180 AP-1-like site (-187 to -181) and the -150 proximal AP-1 site were necessary to observe anergy. Because the -180 site is not required for trans-activation, it was possible to confirm by mutation in the normal mouse IL-2 enhancer that this site is absolutely essential for anergy induction, The simplest model to explain these results is that anergy is mediated by a complex of multiple transcription factors that exert a cis-acting dominant negative regulatory effect on the trans-activation of the IL-2 gene. C1 NIAID,NIH,CELLULAR & MOL IMMUNOL LAB,BETHESDA,MD 20892. NR 49 TC 34 Z9 34 U1 0 U2 0 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD SEP 15 PY 1996 VL 157 IS 6 BP 2328 EP 2339 PG 12 WC Immunology SC Immunology GA VL103 UT WOS:A1996VL10300014 PM 8805630 ER PT J AU Marth, T Strober, W Kelsall, BL AF Marth, T Strober, W Kelsall, BL TI High dose oral tolerance in ovalbumin TCR-transgenic mice - Systemic neutralization of IL-12 augments TGF-beta secretion and T cell apoptosis SO JOURNAL OF IMMUNOLOGY LA English DT Article ID MYELIN BASIC-PROTEIN; GROWTH-FACTOR-BETA; EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS; CHOLERA-TOXIN; ACTIVE SUPPRESSION; HUMAN-LYMPHOCYTES; DENDRITIC CELLS; II COLLAGEN; TH2 CELLS; ANTIGEN AB The immune response to oral Ag administration, including the development of oval tolerance, was explored with the use of OVA TCR-transgenic mice, Feeding high doses of OVA enhanced IFN-gamma production in the Peyer's patches, but induced tolerance in the peripheral lymphoid tissues marked by suppressed proliferative and cytokine responses, Systemic administration of Abs to IL-12 (anti-IL-12) simultaneous with Ag feeding modestly enhanced the degree of tolerance in the peripheral lymphoid tissues, as shown by increased suppression of proliferative responses after in vitro restimulation, and secondary responses in the popliteal lymph nodes following in vivo challenge and in vitro restimulation, Systemic anti-IL-12 treatment was associated with augmented TGF-beta production and T cell apoptosis in both Peyer's patches and peripheral lymphoid tissues, Cell mixing studies and proliferation assays in the presence of anti-TGF-beta provided evidence that the increased suppression of responses induced by anti-IL-12 was due primarily to the secretion of TGF-beta. These findings suggest that IL-12 negatively regulates two of the main mechanisms of oral tolerance, TGF-beta production and clonal deletion via apoptosis, In addition, they suggest that the combination of oral Ag feeding and systemic anti-IL-12 administration may be of benefit in the treatment of autoimmune diseases. C1 NIAID,CLIN INVEST LAB,MUCOSAL IMMUN SECT,NIH,BETHESDA,MD 20892. NR 39 TC 190 Z9 194 U1 0 U2 0 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD SEP 15 PY 1996 VL 157 IS 6 BP 2348 EP 2357 PG 10 WC Immunology SC Immunology GA VL103 UT WOS:A1996VL10300016 PM 8805632 ER PT J AU Rider, LG Hirasawa, N Santini, F Beaven, MA AF Rider, LG Hirasawa, N Santini, F Beaven, MA TI Activation of the mitogen-activated protein kinase cascade is suppressed by low concentrations of dexamethasone in mast cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID AFFINITY IGE RECEPTOR; BASOPHILIC LEUKEMIA-CELLS; CYTOSOLIC PHOSPHOLIPASE A(2); ARACHIDONIC-ACID RELEASE; RAT RBL-2H3 CELLS; TYROSINE PHOSPHORYLATION; PLASMA-MEMBRANE; TRANSCRIPTION FACTOR; SIGNAL-TRANSDUCTION; HISTAMINE-RELEASE AB Antigen stimulation of mast cells via the IgE receptor, Fc epsilon RI, results in recruitment of the cytosolic tyrosine kinases, Lyn and Syk, and the phosphorylation of proteins, We examined the effects of the glucocorticoid dexamethasone on these events in a cultured (RBL-2H3) mast cell line, Nanomolar concentrations of dexamethasone suppressed phosphorylation of proteins that were associated with the activation of the mitogen-activated protein (MAP) kinase/phospholipase A(2) pathway without inhibiting initial events, For example, tyrosine phosphorylation of the subunits of Fc epsilon RI, Lyn, or Syk or of the Ras-guanine nucleotide exchange factor, Vav, was nut suppressed in cells treated with up to 1 mu M dexamethasone. In contrast, phosphorylation of Raf1, MEK1, p42(mapk) and cytosolic phospholipase A(2), as well as the associated increase in MAP kinase activity and release of arachidonic acid, were markedly inhibited in cells treated with as little as 10 nM dexamethasone-a concentration that only partially inhibited hydrolysis of inositol phospholipids or release of secretory granules, Prolonged exposure to dexamethasone also resulted in a partial decrease in expression of MEK1, p42(mapk) and cytosolic phospholipase A(2), which may contribute further to the effects of dexamethasone can this pathway, Activation of the MAP kinase/phospholipase A(2) pathway by the calcium-mobilizing agent thapsigargin was similarly suppressed in dexamethasone-treated cells. These findings suggested that an early step in the pathway, possibly a step immediately before the activation of Raf1, was suppressed by low concentrations of dexamethasone. C1 NHLBI, LAB MOL IMMUNOL, NIH, BETHESDA, MD 20892 USA. NIAMSD, ARTHRIT & RHEUMATISM BRANCH, BETHESDA, MD USA. OI Rider, Lisa/0000-0002-6912-2458 NR 54 TC 81 Z9 81 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 SEP 15 PY 1996 VL 157 IS 6 BP 2374 EP 2380 PG 7 WC Immunology SC Immunology GA VL103 UT WOS:A1996VL10300019 PM 8805635 ER PT J AU Williams, MS Henkart, PA AF Williams, MS Henkart, PA TI Role of reactive oxygen intermediates in TCR-induced death of T cell blasts and hybridomas SO JOURNAL OF IMMUNOLOGY LA English DT Article ID NF-KAPPA-B; ACTIVATION-INDUCED DEATH; OXIDATIVE STRESS; TRANSCRIPTION FACTORS; IMMUNE-RESPONSES; FAS LIGAND; APOPTOSIS; BCL-2; INHIBITION; CYTOTOXICITY AB The functional role of reactive oxygen intermediates (ROI) in activation-induced death of mature T lymphocytes and hybridomas was tested using antioxidants and inhibitors of enzymes that generate oxidants. These agents were shown to inhibit TCR-triggered death in a concentration-dependent manner, suggesting a possible role of ROI in this death, Since the TCR-induced death of both human T blasts and the murine T cell hybridoma 2B4 involve an initial step of TCR-induced Fas ligand (Fast) up-regulation followed by a lethal step induced by Fas cross-linking, both steps were examined separately for ROI dependence, The thiol antioxidants N-acetyl cysteine and glutathione blocked Fas-induced death triggered via cross-linking either by IgM anti-fas or cell-bound Fast, while the other inhibitors of activation-induced death did not block this late lethal step, None of the agents used blocked early events after TCR ligation, as seen by the lack of inhibition of IL-2 secretion or CD69 up-regulation. However, the nonthiol agents that blocked activation-induced death all blocked Fast up-regulation induced by TCR signals in the hybridoma, as measured by a functional assay, Agents inhibiting Fast up-regulation also inhibited activation-induced ROI generation in the hybridoma, as detected by flow cytometry using dihydrorhodamine oxidation, Furthermore, a good correlation was found between the extent of ROI generation and functional Fast expression in 2B4 cells, Thus, while ROI do not appear to act as downstream mediators of apoptotic death induced by steroid or Fas cross-linking in T cells, they are generated by TCR signaling and appear to participate in Fast up-regulation. C1 NCI,EXPT IMMUNOL BRANCH,NIH,BETHESDA,MD 20892. NR 49 TC 76 Z9 76 U1 0 U2 0 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD SEP 15 PY 1996 VL 157 IS 6 BP 2395 EP 2402 PG 8 WC Immunology SC Immunology GA VL103 UT WOS:A1996VL10300022 PM 8805638 ER PT J AU Parkhurst, MR Salgaller, ML Southwood, S Robbins, PF Sette, A Rosenberg, SA Kawakami, Y AF Parkhurst, MR Salgaller, ML Southwood, S Robbins, PF Sette, A Rosenberg, SA Kawakami, Y TI Improved induction of melanoma-reactive CTL with peptides from the melanoma antigen gp100 modified at HLA-A*0201-binding residues SO JOURNAL OF IMMUNOLOGY LA English DT Article ID TUMOR-INFILTRATING LYMPHOCYTES; CLASS-I MOLECULE; METASTATIC MELANOMA; MULTIPLE EPITOPES; PERIPHERAL-BLOOD; ANCHOR RESIDUES; T-LYMPHOCYTES; IDENTIFICATION; HLA; IMMUNOTHERAPY AB Recognition of the melanoma Ag gp100 by tumor-infiltrating lymphocytes (TIL) in vitro has been correlated with tumor regression in patients with metastatic melanoma treated with the adoptive transfer of TIL plus IL-2. Three common gp100 epitopes have been identified that are recognized in the context of HLA-A2 by TIL from different patients: G9(154) (KTWCQYWQV), C9(209) (ITDQVPFSV), and C9(280) (YLEPGPVTA). Upon stimulation with these peptides, melanoma-reactive CTL could be induced in vitro from PBL of some HLA-A2(+) melanoma patients, However, numerous restimulations were required, and specific reactivity could not be generated in many patients, Therefore, to enhance the immunogenicity of gp100 peptides, amino acid substitutions were introduced into G9(154), G9(209), and G9(280) at HLA-A*0201-binding anchor positions, but not at TCR contact residues, to increase peptide class I MHC-binding affinity. Several modified gp100 peptides bound with greater affinity to HLA-A*0201 than unmodified peptides and were recognized by TIL specific for the natural epitopes, These peptides were used to sensitize PBL from HLA-A2(+) melanoma patients in vitro using peptide-pulsed autologous PBMC as stimulators, After five weekly restimulations with either the native C9(209) or G9(280) peptide, melanoma-reactive CTL could only be induced from two of seven patients, However, amino acid substitutions in these peptides enabled the induction of melanoma-reactive CTL from all seven patients, These results suggest that modified gp100 peptides may be more immunogenic than the native epitopes, and may be useful in immunotherapy protocols for patients with melanoma. C1 CYTEL CORP,DEPT IMMUNOL,SAN DIEGO,CA 92121. RP Parkhurst, MR (reprint author), NCI,SURG BRANCH,NIH,BLDG 10,RM 2B42,10 CTR DR MSC 1502,BETHESDA,MD 20892, USA. RI Kawakami, Yutaka /E-7429-2013 OI Kawakami, Yutaka /0000-0003-4836-2855 NR 50 TC 445 Z9 453 U1 0 U2 5 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD SEP 15 PY 1996 VL 157 IS 6 BP 2539 EP 2548 PG 10 WC Immunology SC Immunology GA VL103 UT WOS:A1996VL10300039 PM 8805655 ER PT J AU Chen, HJ Kinzer, CA Paul, WE AF Chen, HJ Kinzer, CA Paul, WE TI p161, a murine membrane protein expressed on mast cells and some macrophages, is mouse CD13/aminopeptidase N SO JOURNAL OF IMMUNOLOGY LA English DT Article ID AMINO-ACID-SEQUENCE; ACUTE LYMPHOBLASTIC-LEUKEMIA; AMINOPEPTIDASE-N; MESSENGER-RNA; MONOCLONAL-ANTIBODIES; ANTIGEN EXPRESSION; MYELOID CELLS; SURFACE; RECEPTOR; CD13 AB p161 is a membrane glycoprotein expressed on mast cells and on activated macrophages but on few if any other cells of hematopoietic lineages, Its lack of expression on basophils makes it useful to distinguish mast cells from basophils and aids in the analysis of mast cells and their precursors, p161 was purified from the mast cell line CFTL-12 by affinity chromatography and subjected to limited proteolysis, The sequences of the resultant peptides indicated that p161 is homologous with rat and human CD13/aminopeptidase N, Using oligonucleotide primers derived from rat CD13 cDNA, a mouse cDNA was obtained, Its deduced amino acid sequence displays 87% identity with rat CD13 and 76% identity with human CD13, Expression of the mouse cDNA in M12 cells, which are p161 negative, renders these cells positive for staining with the monoclonal anti-p161 Ab, K-1, Furthermore, a mAb raised against partially purified mouse intestinal aminopeptidase N specifically blocked the binding of K-1 to both CFTL-12 cells and the transfected M12 cells, These results strongly indicate that mouse p161 is CD13/aminopeptidase N. Northern blot analysis shows that p161 mRNA is most abundantly expressed in the intestinal tract and kidney and is present in liver, lymph node, spleen, and brain. C1 NIAID,IMMUNOL LAB,NIH,BETHESDA,MD 20892. NR 50 TC 21 Z9 22 U1 0 U2 0 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD SEP 15 PY 1996 VL 157 IS 6 BP 2593 EP 2600 PG 8 WC Immunology SC Immunology GA VL103 UT WOS:A1996VL10300046 PM 8805662 ER PT J AU Melillo, G Taylor, LS Brooks, A Cox, GW Varesio, L AF Melillo, G Taylor, LS Brooks, A Cox, GW Varesio, L TI Regulation of inducible nitric oxide synthase expression in IFN-gamma-treated murine macrophages cultured under hypoxic conditions? SO JOURNAL OF IMMUNOLOGY LA English DT Article ID MOUSE MACROPHAGES; PICOLINIC-ACID; L-ARGININE; TRANSCRIPTION FACTOR; INTERFERON-GAMMA; ERYTHROPOIETIN GENE; ALLOGRAFT-REJECTION; MAMMALIAN-CELLS; OXYGEN-TENSION; GROWTH-FACTOR AB We recently reported that a hypoxia-responsive element mediates a novel pathway of transcriptional activation of the inducible nitric oxide synthase (iNOS) promoter in murine macrophages treated with IFN gamma plus hypoxia (1% O-2). In this study, we investigated the expression of iNOS activity and the regulation of iNOS induction in IFN-gamma-treated ANA-1 murine macrophages or thioglycollate-elicited peritoneal macrophages cultured under hypoxic conditions. We found that murine macrophages stimulated with IFN-gamma plus hypoxia, despite a significant accumulation of iNOS mRNA, did not release nitrite into culture supernatant. However, cytosol from macrophages treated with IFN-gamma plus hypoxia contained significant levels of iNOS protein and enzymatic activity. Experiments in which cells were treated with IFN-gamma plus hypoxia and then cultured in normoxic conditions (20% O-2) demonstrated that reoxygenation was required to achieve detectable accumulation of nitrite in the culture supernatant. Furthermore, we demonstrated that lL-4 inhibited IFN-gamma plus hypoxia-dependent induction of iNOS mRNA expression, iNOS protein, and enzymatic activity. Experiments in which ANA-1 macrophages were transfected transiently with the full-length iNOS promoter linked to a chloramphenicol acetyltransferase reporter gene demonstrated that IL-4 also downregulated the IFN-gamma plus hypoxia-induced activation of the iNOS promoter. These data establish that hypoxia is a costimulus with IFN-gamma for the induction of iNOS activity in ANA-1 macrophages as well as in murine peritoneal macrophages, and they provide the first evidence that IL-4 inhibits hypoxia-inducible gene expression, In addition, our results suggest that hypoxia,which occurs in many pathologic conditions, may play an important role in the activation of murine macrophages. C1 NCI,FREDERICK CANC RES FACIL,INTRAMURAL RES SUPPORT PROGRAM,SAIC FREDERICK,FREDERICK,MD 21702. RP Melillo, G (reprint author), NCI,FREDERICK CANC RES FACIL,EXPT IMMUNOL LAB,DIV BASIC SCI,BLDG 560,RM 31-52,FREDERICK,MD 21702, USA. RI varesio, luigi/J-8261-2016 OI varesio, luigi/0000-0001-5659-2218 NR 56 TC 87 Z9 87 U1 0 U2 1 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD SEP 15 PY 1996 VL 157 IS 6 BP 2638 EP 2644 PG 7 WC Immunology SC Immunology GA VL103 UT WOS:A1996VL10300052 PM 8805668 ER PT J AU Caspi, RR Silver, PB Chan, CC Sun, B Agarwal, RK Wells, J Oddo, S Fujino, Y Najafian, F Wilder, RL AF Caspi, RR Silver, PB Chan, CC Sun, B Agarwal, RK Wells, J Oddo, S Fujino, Y Najafian, F Wilder, RL TI Genetic susceptibility to experimental autoimmune uveoretinitis in the rat is associated with an elevated Th1 - Response SO JOURNAL OF IMMUNOLOGY LA English DT Article ID T-CELLS; MHC AB This study examines whether genetic susceptibility vs genetic resistance to experimental autoimmune uveoretinitis (EAU) are connected to a predisposition to mount a Th1-dominated (IFN-gamma high, IL-4 low) vs a Th2-dominated (IL-4 high, IFN-gamma low) response, Lewis rats developed disease with high incidence after immunization with the uveitogenic peptide R16, whereas F344 rats were resistant. Primed lymph node cells from both strains proliferated in culture in response to R16, However, while the Lewis cultures transferred EAU to syngeneic recipients, those of F344 did not, The Lewis cultures produced substantially more IFN-gamma mRNA and protein in response to R16, than did those of F344, Both strains made low levels of IL-10 mRNA and IL-4 mRNA. Unlike the primary cultures, long-term (R16-specific) T cell lines derived from each of the strains transferred EAU equally well to their respective recipients, and produced similar, high levels of IFN-gamma mRNA and protein, Treatment of F344 with Bordefella pertussis toxin concurrently with immunization abrogated its resistance,enhanced Ag-specific IFN-gamma production in culture, and yielded a primed cell population capable of transferring EAU, Conversely, immunization of Lewis rats with R16 in IFA induced little or no disease; the primed cells produced minimal amounts of IFN-gamma and did not transfer EAU. However, addition of IL-12 into the culture resulted in a highly pathogenic, IFN-gamma-producing cell population, We conclude that genetic susceptibility to ocular autoimmunity in this model is connected to an elevated Th1 response, Genetic resistance, however, does not seem to involve an elevated Th2 response, but rather an inhibited development of Th1-like effector cells. C1 NIAMSD, NATL INST HLTH, BETHESDA, MD 20892 USA. RP Caspi, RR (reprint author), NEI, IMMUNOL LAB,NIH,BLDG 10,ROOM 10N222, 10 CTR DR, MSC 1858, BETHESDA, MD 20892 USA. NR 23 TC 105 Z9 113 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 SEP 15 PY 1996 VL 157 IS 6 BP 2668 EP 2675 PG 8 WC Immunology SC Immunology GA VL103 UT WOS:A1996VL10300056 PM 8805672 ER PT J AU Szekely, Z Konya, Z Becskei, A Goldring, WPD Perczel, A Penke, B Molnar, J Michejda, CF Aszalos, A Csizmadia, IG AF Szekely, Z Konya, Z Becskei, A Goldring, WPD Perczel, A Penke, B Molnar, J Michejda, CF Aszalos, A Csizmadia, IG TI Suggested binding mechanism of the HIV-gp120 to its CD4 receptor SO JOURNAL OF MOLECULAR STRUCTURE-THEOCHEM LA English DT Article DE ab initio calculation; anti-HIV drug; CD4; gp120; docking; MM and MD simulations ID HUMAN-IMMUNODEFICIENCY-VIRUS; ENVELOPE GLYCOPROTEIN GP120; MOLECULAR-ORBITAL METHODS; VALENCE BASIS-SETS; CRYSTAL-STRUCTURE; ATOMIC-STRUCTURE; PEPTIDE MODELS; SOLUBLE CD4; RAT CD4; HIV AB The molecular recognition and attachment of the CD4 molecule and the HIV envelope glycoprotein (gp120) might be described as a consecutive three-step molecular recognition process. (a) Long range interaction: electrostatic pre-orientation, (b) short range interaction: electronic attachment followed by a 'Locking-in' (via aromatic ring orientation) and (c) internal interaction (induced fit): conformational readjustment of the protein molecules. On the basis of the preliminary investigations (X-ray structures of CD4 and biological studies of CD4 and gp120 point mutants) we described a computational model. This approach consists of empirical calculations as well as ab initio level of quantum chemistry. The conformational analysis of the wild type and mutant CD4 molecules was supported by molecular mechanics and dynamics (Amber force field). The latter analysis involves the application of a novel method, the Amino Acid Conformation Assignment of Proteins (ACAP) software, developed for the notation of secondary protein structures. According to the cardinal role of the electrostatic factors during this interaction, several ab initio investigations were performed for better understanding of the recognition process on submolecular level. Using the above mentioned computational model, we could interpret the basic behaviours and predict some additional features of CD4-gp120 interaction, in spite of the missing gp120 X-ray structure. C1 UNIV TORONTO, DEPT CHEM, TORONTO, ON M5S 1A1, CANADA. NCI, FREDERICK CANC RES & DEV CTR, ABL BASIC RES PROGRAM,MSL, MOL ASPECTS DRUG DESIGN SECT, FREDERICK, MD 21702 USA. ATTILA JOZSEF UNIV, DEPT APPL CHEM, H-6720 SZEGED, HUNGARY. EOTVOS LORAND UNIV, DEPT ORGAN CHEM, H-1518 BUDAPEST, HUNGARY. UNIV OXFORD, DEPT BIOCHEM, OXFORD OX1 3QU, ENGLAND. ALBERT SZENT GYORGYI MED UNIV, DEPT MICROBIOL, H-6720 SZEGED, HUNGARY. US FDA, CELL BIOL LABS, WASHINGTON, DC 20204 USA. RP Szekely, Z (reprint author), ALBERT SZENT GYORGYI MED UNIV, DEPT MED CHEM, DOM TER 8, H-6720 SZEGED, HUNGARY. RI Konya, Zoltan/C-2492-2009; OI Konya, Zoltan/0000-0002-9406-8596; Becskei, Attila/0000-0002-9462-0934 NR 41 TC 3 Z9 3 U1 1 U2 3 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0166-1280 J9 J MOL STRUC-THEOCHEM JI Theochem-J. Mol. Struct. PD SEP 15 PY 1996 VL 367 BP 159 EP 186 DI 10.1016/S0166-1280(96)04501-0 PG 28 WC Chemistry, Physical SC Chemistry GA VV061 UT WOS:A1996VV06100019 ER PT J AU Levine, MS Altemus, KL Cepeda, C Cromwell, HC Crawford, C Ariano, MA Drago, J Sibley, DR Westphal, H AF Levine, MS Altemus, KL Cepeda, C Cromwell, HC Crawford, C Ariano, MA Drago, J Sibley, DR Westphal, H TI Modulatory actions of dopamine on NMDA receptor-mediated responses are reduced in D-1A-deficient mutant mice SO JOURNAL OF NEUROSCIENCE LA English DT Article DE dopamine receptors; D-1; excitatory amino acid receptors; knock-out mice; mutant; neostriatal slices; NMDA ID EXCITATORY AMINO-ACIDS; RAT STRIATAL NEURONS; CAT CAUDATE NEURONS; NEOSTRIATAL NEURONS; SYNAPTIC TRANSMISSION; CORTICAL INPUTS; MIXED SYNAPSES; BASAL GANGLIA; IN-VITRO; INVITRO AB The role of D-1 dopamine (DA) receptors in mediating the ability of DA to modulate responses attributable to activation of NMDA receptors was examined in mice lacking D-1A dopamine receptors. Specifically, experiments were designed to test the hypothesis that the ability of DA to potentiate responses mediated by activation of NMDA receptors was attributable to activation of D-1 receptors. Based on this hypothesis, we would predict that in the D-1A mutant mouse, either DA would not induce enhancement of NMDA-mediated responses, or the enhancement would be severely attenuated. The results provided evidence to support the hypothesis. In mutant mice, DA and D-1 receptor agonists did not potentiate responses mediated by activation of NMDA receptors. In contrast, in control mice, both DA and D-1 receptor agonists markedly potentiated responses mediated by activation of NMDA receptors. The effects of DA in attenuating responses mediated by activation of non-NMDA receptors also were altered in the mutant, suggesting that this action of DA may require coupling or interactions between D-1 and D-2 receptors. The present studies also provided an opportunity to assess some of the basic electrophysiological and morphological properties of neostriatal neurons in mice lacking D-1A DA receptors. Resting membrane potential, action potential parameters, input resistance, excitability, somatic size, dendritic extent, and estimates of spine density in mutants and controls were similar, suggesting that these basic neurophysiological and structural properties have not been changed by the loss of the D-1A DA receptor. C1 FINCH UNIV HLTH SCI CHICAGO MED SCH,DEPT NEUROSCI,N CHICAGO,IL 60064. MONASH UNIV,DEPT ANAT,CLAYTON,VIC 3168,AUSTRALIA. NINCDS,EXPT THERAPEUT BRANCH,BETHESDA,MD 20892. NICHHD,LAB MAMMALIAN GENES & DEV,BETHESDA,MD 20892. RP Levine, MS (reprint author), UNIV CALIF LOS ANGELES,MENTAL RETARDAT RES CTR,760 WESTWOOD PLAZA,LOS ANGELES,CA 90024, USA. RI Crawford, cynthia/G-2603-2012; OI Cromwell, Howard/0000-0003-0464-7082 FU NIA NIH HHS [AG 10252]; NICHD NIH HHS [HD05958]; NINDS NIH HHS [NS 35233] NR 53 TC 143 Z9 143 U1 0 U2 1 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD SEP 15 PY 1996 VL 16 IS 18 BP 5870 EP 5882 PG 13 WC Neurosciences SC Neurosciences & Neurology GA VG900 UT WOS:A1996VG90000027 PM 8795639 ER PT J AU Tranquill, LR Skinner, E Campagnoni, C Vergelli, M Hemmer, B Muraro, P Martin, R McFarland, HF Campagnoni, AT Voskuhl, RR AF Tranquill, LR Skinner, E Campagnoni, C Vergelli, M Hemmer, B Muraro, P Martin, R McFarland, HF Campagnoni, AT Voskuhl, RR TI Human T lymphocytes specific for the immunodominant 83-99 epitope of myelin basic protein: Recognition of golli MBP HOG 7 SO JOURNAL OF NEUROSCIENCE RESEARCH LA English DT Article DE golli myelin basic protein; T lymphocyte; multiple sclerosis ID TUMOR-NECROSIS-FACTOR; CENTRAL NERVOUS SYSTEMS; CELL RECEPTOR USAGE; MULTIPLE-SCLEROSIS; PROTEOLIPID PROTEIN; CEREBROSPINAL-FLUID; FINE SPECIFICITY; IMMUNE-SYSTEM; FACTOR-ALPHA; EXPRESSION AB Myelin basic protein (MBP) is a candidate autoantigen in the disease multiple sclerosis, Although MBP was thought to be sequestered behind the blood-brain barrier, isoforms of MBPs have recently been demonstrated in lymphoid tissues, These isoforms, termed golli MBPs, contain sequences that are shared with ''classic'' MBP within the CNS. In the present study, we have determined that epitopes within golli MBP isoforms may be recognized by human T lymphocyte clones specific for classic MBP, Ten of 12 T-cell clones recognized golli MBP, Although 11 clones were specific for the immunodominant 83-99 sequence, the clones differed with respect to human leukocyte antigen (HLA) restriction. T-helper phenotype, cytolytic activity, and T-cell receptor usage, Greater responses to classic MBP than to golli MBP suggested a difference in the ability of the two proteins to be processed and to present epitopes therein. These data advance the hypothesis that golli MBP sequences expressed within lymphoid tissues may be recognized by classic MBP-specific T lymphocytes during central or peripheral tolerance. (C) 1996 Wiley-Liss, Inc. C1 UNIV CALIF LOS ANGELES,SCH MED,DEPT NEUROL,LOS ANGELES,CA 90095. UNIV CALIF LOS ANGELES,SCH MED,MENTAL RETARDAT RES CTR,LOS ANGELES,CA 90095. NIH,NEUROIMMUNOL BRANCH,BETHESDA,MD 20892. FU NINDS NIH HHS [NS33091] NR 47 TC 18 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 SN 0360-4012 J9 J NEUROSCI RES JI J. Neurosci. Res. PD SEP 15 PY 1996 VL 45 IS 6 BP 820 EP 828 DI 10.1002/(SICI)1097-4547(19960915)45:6<820::AID-JNR19>3.0.CO;2-Z PG 9 WC Neurosciences SC Neurosciences & Neurology GA VK550 UT WOS:A1996VK55000019 PM 8892094 ER PT J AU Hemmer, B Vergelli, M Calabresi, P Huang, T McFarland, HF Martin, R AF Hemmer, B Vergelli, M Calabresi, P Huang, T McFarland, HF Martin, R TI Cytokine phenotype of human autoreactive T cell clones specific for the immunodominant myelin basic protein peptide (83-99) SO JOURNAL OF NEUROSCIENCE RESEARCH LA English DT Article DE cytokine; MS; MBP; EAE ID EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS; CD4+ SUPPRESSOR CELLS; NECROSIS-FACTOR-ALPHA; MULTIPLE-SCLEROSIS; AUTOIMMUNE ENCEPHALOMYELITIS; LYMPHOCYTE-RESPONSES; PROTEOLIPID PROTEIN; INTERFERON-GAMMA; PERIPHERAL-BLOOD; EFFECTOR-CELLS AB Experimental allergic encephalomyelitis (EAE), an animal model resembling multiple sclerosis (MS), is mediated by myelin antigen-specific CD4+ T cells secreting cytokines such as interferon-gamma (IFN-gamma), tumor necrosis factor-beta (TNF-beta), and the proinflammatory cytokine TNF-alpha-all associated with the T-helper-1 (Th1) T cell subset, Based on numerous similarities between MS and EAE, it has been postulated that Th1-like T cells are involved in the pathogenesis of MS, Production of proinflammatory cytokines such as IFN-gamma and, in particular; TNF-alpha/beta by autoreactive T cells is considered crucial for the initiation and amplification of inflammatory brain lesions and possibly also for direct myelin damage, In contrast, regulatory cytokines such as interleukin-4 (IL-4), IL-10, and IL-13, which are associated with the Th2-like phenotype, may play a role in the resolution of relapses, Although the human T cell response to myelin basic protein (MBP) is well characterized in terms of antigen specificity, HLA restriction, and T cell-receptor (TCR) usage, little is known about the cytokine pattern of these autoreactive T cells. To gain such information, conditions for studying cytokine secretion by human autoreactive T cell clones (TCC) were established, The cytokine secretion profile of human autoreactive CD4+ TCC, specific for myelin basic protein peptide (83-89) [MBP(83-99)], a candidate autoantigen in MS, was investigated. Our results show that TCC cytokine production in long-term culture was stable. In addition, the correlation of various cytokines within specific TCC revealed differences compared to murine T cells. The comparison of 30 human MBP(83-99)-specific TCC demonstrated heterogeneity in cytokine secretion, with a continuum between Th1- and Th2-like cells rather than distinct Th1 or Th2 subsets, These data are important for further investigation of the potential role of cytokines in the inflammatory process of MS, and provide a powerful tool to investigate therapeutic interventions with respect to their influence on cytokine secretion of autoreactive T cells. (C) 1996 Wiley-Liss, Inc. C1 NINCDS,NEUROIMMUNOL BRANCH,NIH,BETHESDA,MD 20892. UNIV TUBINGEN,SCH MED,DEPT NEUROL,TUBINGEN,GERMANY. FU NINDS NIH HHS [1R43NS33063-01] NR 42 TC 43 Z9 43 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC 605 THIRD AVE, NEW YORK, NY 10158-0012 SN 0360-4012 J9 J NEUROSCI RES JI J. Neurosci. Res. PD SEP 15 PY 1996 VL 45 IS 6 BP 852 EP 862 PG 11 WC Neurosciences SC Neurosciences & Neurology GA VK550 UT WOS:A1996VK55000022 PM 8892097 ER PT J AU Gerthoffer, WT Yamboliev, IA Shearer, M Pohl, J Haynes, R Dang, S Sato, K Sellers, JR AF Gerthoffer, WT Yamboliev, IA Shearer, M Pohl, J Haynes, R Dang, S Sato, K Sellers, JR TI Activation of MAP kinases and phosphorylation of caldesmon in canine colonic smooth muscle SO JOURNAL OF PHYSIOLOGY-LONDON LA English DT Article ID MYOSIN LIGHT CHAIN; PROTEIN-KINASE; ACTIN-FILAMENTS; PHORBOL ESTER; H-CALDESMON; IDENTIFICATION; MOVEMENT; EXPRESSION; SEQUENCES; BINDING AB 1. Phosphorylation of caldesmon was assayed in canine colonic circular smooth muscle strips labelled with P-32 and stimulated with 10 mu M acetylcholine. Caldesmon was isolated by two-dimensional non-equilibrium pH gel electrophoresis. Stimulation with acetylcholine increased caldesmon phosphorylation significantly from a basal level of 0.6 +/- 0.07 to 1.1 +/- 0.15 mol P-i (mol caldesmon)(-1) after 2 min. 2. MAP kinase activities were measured in SDS extracts of muscle by a gel reconstitution method using myelin basic protein. Myelin basic protein kinase activities were observed at 38, 44, 50 and 57 kDa by the gel reconstitution method. Endogenous caldesmon kinase activities were also identified by the gel reconstitution method at 38, 44 and 50 kDa. The 38 and 44 kDa kinases comigrated with proteins labelled by anti-ERK1 MAP liinase antibodies on Western blots. Both 38 and 44 kDa MBP kinase activities increased significantly during contractions induced by 10 mu M acetylcholine, 0.1 mu M neurokinin A and 70 mM potassium. 3. Phorbol dibutyrate (0.1 mu M) potentiated activation of MAP kinases and contraction of depolarized muscles while producing a decrease in fura-2 fluorescence ratio. This suggests that protein liinase C activation is coupled to MAP kinase activity in colonic smooth muscle. 4. MAP kinases isolated from muscle homogenates by Mono & chromatography were assayed using the specific MAP kinase substrate peptide APRTPGGRR. Stimulation of muscles for 2 min with 10 mu M acetylcholine activated both ERK1 and ERK2 MAP kinase activities 2-fold. 5. To determine the effects of caldesmon phosphorylation by MAP kinase on the cross-bridge cycle, actin sliding velocity was measured with an in vitro motility assay, Unphosphorylated turkey gizzard caldesmon (3 mu M) significantly reduced mean sliding velocity. Phosphorylation of caldesmon with sea star ERK1 MAP kinase reversed the inhibitory effect of caldesmon on sliding velocity. The results are consistent with a protein kinase cascade being activated by contractile agonists in gastrointestinal smooth muscle which activates ERK MAP kinases leading to phosphorylation of caldesmon. Phosphorylation of caldesmon in viveo may reverse inhibitory influences of caldesmon on cross-bridge cycling. C1 UNIV TOKYO, FAC AGR, RADIO ISOTOPE CTR, BUNKYO KU, TOKYO 113, JAPAN. NHLBI, MOL CARDIOL LAB, NIH, BETHESDA, MD 20892 USA. RP UNIV NEVADA, SCH MED, DEPT PHARMACOL, RENO, NV 89557 USA. OI Gerthoffer, William/0000-0001-5154-4962 FU NHLBI NIH HHS [HL48183]; NIDDK NIH HHS [DK41315] NR 38 TC 95 Z9 95 U1 0 U2 0 PU WILEY-BLACKWELL PI HOBOKEN PA 111 RIVER ST, HOBOKEN 07030-5774, NJ USA SN 0022-3751 EI 1469-7793 J9 J PHYSIOL-LONDON JI J. Physiol.-London PD SEP 15 PY 1996 VL 495 IS 3 BP 597 EP 609 PG 13 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA VL090 UT WOS:A1996VL09000001 PM 8887769 ER PT J AU Rosenberg, HF Dyer, KD AF Rosenberg, HF Dyer, KD TI Molecular cloning and characterization of a novel human ribonuclease (RNase k6): Increasing diversity in the enlarging ribonuclease gene family SO NUCLEIC ACIDS RESEARCH LA English DT Article ID EOSINOPHIL-DERIVED NEUROTOXIN; AMINO-ACID-SEQUENCE; POLYMERASE CHAIN-REACTION; CATIONIC PROTEIN; PANCREATIC RIBONUCLEASE; SEMINAL RIBONUCLEASE; HUMAN ANGIOGENIN; RANA-CATESBEIANA; MESSENGER-RNA; SUPERFAMILY AB The discovery of Ribonuclease k6 (RNase k6) was an unexpected result of our ongoing efforts to trace the evolutionary history of the ribonuclease gene family. The open reading frame of RNase k6, amplified from human genomic DNA, encodes a 150 amino acid polypeptide with eight cysteines and histidine and lysine residues corresponding to those found in the active site of the prototype, ribonuclease A. The single-copy gene encoding RNase k6 maps to human chromosome 14 and orthologous sequences were detected in both primate and non-primate mammalian species. A single mRNA transcript (1.5 kb) was detected in all human tissues tested, with lung representing the most abundant source. At the cellular level, transcripts encoding RNase k6 were detected in normal human monocytes and neutrophils (but not in eosinophils) suggesting a role for this ribonuclease in host defense. Of the five previously identified human ribonucleases of this group, RNase k6 is most closely related to eosinophil-derived neurotoxin (EDN), with 47% amino acid sequence identity; slight crossreactivity between RNase k6 and EDN was observed on Western blots probed with polyclonal anti-EDN antiserum. The catalytic constants determined, K-m = 5.0 mu M and k(cat) = 0.13 s(-1), indicate that recombinant RNase k6 has similar to 40-fold less ribonuclease activity than recombinant EDN. The identification and characterization of RNase k6 has extended the ribonuclease gene family and suggests the possibility that there are others awaiting discovery. RP Rosenberg, HF (reprint author), NIAID,HOST DEF LAB,NIH,9000 ROCKVILLE PIKE,BETHESDA,MD 20892, USA. NR 53 TC 53 Z9 56 U1 1 U2 2 PU OXFORD UNIV PRESS UNITED KINGDOM PI OXFORD PA WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP SN 0305-1048 J9 NUCLEIC ACIDS RES JI Nucleic Acids Res. PD SEP 15 PY 1996 VL 24 IS 18 BP 3507 EP 3513 DI 10.1093/nar/24.18.3507 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VK032 UT WOS:A1996VK03200004 PM 8836175 ER PT J AU Maraia, R Sakulich, AL Brinkmann, E Green, ED AF Maraia, R Sakulich, AL Brinkmann, E Green, ED TI Gene encoding human Ro-associated autoantigen Y5 RNA SO NUCLEIC ACIDS RESEARCH LA English DT Article ID SMALL CYTOPLASMIC RIBONUCLEOPROTEINS; POLYMERASE-III TRANSCRIPTION; SMALL NUCLEAR-RNA; 5S RIBOSOMAL-RNA; BINDING PROTEIN; ALU REPEATS; LUPUS-ERYTHEMATOSUS; HUMAN GENOME; SEQUENCE; SITE AB Ro ribonucleoproteins are composed of Y RNAs and the Ro 60 kDa protein. While the Ro 60 kDa protein is implicated in an RNA discard pathway that recognizes 3'-extended 5S rRNAs, the function of Y RNAs remains unknown [O'Brien, C.A. and Wolin, S.L. (1995) Genes Dev. 8, 2891-2903], Y5 RNA occupies a large fraction of Ro 60 kDa protein in human Ro RNPs, contains an atypical 3'-extension not found on other Y RNAs, and constitutes an RNA antigen in certain autoimmune patients [Boulanger et al, (1995) Clin. Exp. Immunol. 99, 29-36]. An overabundance of Y RNA retroposed pseudogenes has previously complicated the isolation of mammalian Y RNA genes. The source gene for Y5 RNA was isolated from human DNA as well as from Galago senegalis DNA. Authenticity of the hY5 RNA gene was demonstrated in vivo and its activity was compared with the hY4 RNA gene that also uses a type 3 promoter for RNA polymerase III. The hY5 RNA gene was subsequently found to reside within a few hundred thousand base pairs of other Y RNA genes and the linear order of the four human Y RNA genes on chromosome 7q36 was determined. Phylogenetic comparative analyses of promoter and RNA structure indicate that the Y5 RNA gene has been subjected to positive selection during primate evolution. Consistent with the proposal of O'Brien and Harley [O'Brian, C.A. and Wolin, S.L. (1992) Gene 116, 285-289], analysis of flanking sequences suggest that the hY5 RNA gene may have originated as a retroposon. C1 NATL CTR HUMAN GENOME RES,DIAGNOST DEV BRANCH,NIH,BETHESDA,MD 20892. RP Maraia, R (reprint author), NICHHD,MOL GROWTH REGULAT LAB,9000 ROCKVILLE PIKE,BETHESDA,MD 20892, USA. NR 51 TC 23 Z9 23 U1 0 U2 2 PU OXFORD UNIV PRESS UNITED KINGDOM PI OXFORD PA WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP SN 0305-1048 J9 NUCLEIC ACIDS RES JI Nucleic Acids Res. PD SEP 15 PY 1996 VL 24 IS 18 BP 3552 EP 3559 DI 10.1093/nar/24.18.3552 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VK032 UT WOS:A1996VK03200011 PM 8836182 ER PT J AU Kobayashi, K Ohye, T Pastan, I Nagatsu, T AF Kobayashi, K Ohye, T Pastan, I Nagatsu, T TI A novel strategy for the negative selection in mouse embryonic stem cells operated with immunotoxin-mediated cell targeting SO NUCLEIC ACIDS RESEARCH LA English DT Article ID HOMOLOGOUS RECOMBINATION; TRANSGENIC MICE; GENE; DISRUPTION; MUTATIONS; EXPRESSION; LOCUS AB Immunotoxoin-mediated cell targeting (IMCT)isa technique for conditionally ablating specific cell types based on the cytotoxic activity of a recombinant immunotoxin anti-Tac (Fv)-PE40. To examine the feasibility of this technique for the negative selection in mouse embryonic stem (ES) cells, we investigated the responsiveness of cells expressing human interleukin-2 receptor alpha subunit to anti-Tac(Fv)-PE40. The immunotoxin treatment efficiently eliminated only ES cells bearing the receptor as a consequence of the target specificity of anti-Tac(Fv)-PE40, indicating that IMCT can be used as a novel strategy for positive and negative selection to enrich ES cell clones with a targeted mutation. C1 FUJITA HLTH UNIV,SCH MED,INST COMPREHENS MED SCI,TOYOAKE,AICHI 47011,JAPAN. NARA INST SCI & TECHNOL,RES & EDUC CTR GENET INFORMAT,NARA 63001,JAPAN. NCI,NIH,MOL BIOL LAB,DIV BASIC SCI,BETHESDA,MD 20892. NR 13 TC 4 Z9 4 U1 0 U2 0 PU OXFORD UNIV PRESS UNITED KINGDOM PI OXFORD PA WALTON ST JOURNALS DEPT, OXFORD, ENGLAND OX2 6DP SN 0305-1048 J9 NUCLEIC ACIDS RES JI Nucleic Acids Res. PD SEP 15 PY 1996 VL 24 IS 18 BP 3653 EP 3655 DI 10.1093/nar/24.18.3653 PG 3 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VK032 UT WOS:A1996VK03200029 PM 8836200 ER PT J AU Nossal, R AF Nossal, R TI Mechanical properties of biological gels SO PHYSICA A LA English DT Article; Proceedings Paper CT Workshop on Colloid and Interface Science - Trends and Applications, on the Occasion of the Celebration of the 60th Birthday of Sow-Hsin Chen CY MAY 02-05, 1995 CL GUANICA, PR SP ES, PSCoR NSF, USN, Off Naval Res ID NETWORKS; ACTIN; CYTOSKELETON AB The rheological properties of biological gels are illustrated by reviewing measurements made on samples containing polymer networks reconstituted from cytoskeletal constituents. Although these networks are crosslinked by non-covalent interactions which may lead to transient gel behavior, the early-time (rapid) mechanical response of the gels can be characterized by the instantaneous shear modulus. A theory for the latter is described, and used to examine the relationship between the cortical f-actin content and the stiffening of activated polymorphonuclear leukocytes. RP Nossal, R (reprint author), NIH,PHYS SCI LAB,DIV COMP RES & TECHNOL,BLDG 12A,ROOM 2007,BETHESDA,MD 20892, USA. NR 22 TC 1 Z9 1 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-4371 J9 PHYSICA A JI Physica A PD SEP 15 PY 1996 VL 231 IS 1-3 BP 265 EP 276 DI 10.1016/0378-4371(95)00455-6 PG 12 WC Physics, Multidisciplinary SC Physics GA VH367 UT WOS:A1996VH36700022 ER PT J AU Maciejewski, JP Young, NS Yu, M Anderson, SM Sloand, EM AF Maciejewski, JP Young, NS Yu, M Anderson, SM Sloand, EM TI Analysis of the expression of glycosylphosphatidylinositol anchored proteins on platelets from patients with paroxysmal nocturnal hemoglobinuria SO THROMBOSIS RESEARCH LA English DT Article DE paroxysmal nocturnal hemoglobinuria (PNH); platelet; glycosylphosphatidylinositol; thrombosis; hemolysis ID MEMBRANE INHIBITOR; REACTIVE LYSIS; COMPLEMENT; GLYCOPROTEINS; ERYTHROCYTES; CLONALITY; COMPLEXES; SURVIVAL; SURFACE; CELLS AB Paroxysmal nocturnal hemoglobinuria (PNH), an acquired clonal disorder is manifested by failure of hematopoietic cells to express phosphatidylinositol glycan-anchored proteins (PIG-AP). Since the PIG-A mutation is present at the stem cell level, all cell lines may be affected. Although the pathogenesis of hemolytic anemia in PNH is related to the absence of CD55 and CD59 molecules on the membrane of red cells, the mechanism responsible for the increased incidence of thrombotic events in PNH is not clear. In this study we measured two glycosylphosphatidylinositol (GPI)-linked molecules on platelets (CD55 and CD59) and two GPI-linked proteins on neutrophils (CD14 and CD16), comparing their expression on normal and PNH patients. Using two-color flow cytometric analysis with antibodies directed against CD42b and CD41a, we found that CD55 and CD59 were constitutively expressed by normal fresh platelets, but that the expression levels decreased during the five day storage of platelets. A substantial population of platelets lacking the GPI-linked proteins were detected in most cases. We demonstrated varying degrees of deficiency in the expression of GPI-anchored molecules with neutrophils, monocytes and platelets with the highest proportion of deficient cells found within monocytic lineage. Similar numbers of platelets with the PNH phenotype and normal platelets expressed activation markers before and after exposure to platelet agonists. Flow cytometry is more sensitive than Ham's test in monitoring expression of PNH in platelets. Differences in the numbers of-circulating GPI-deficient platelets and myeloid cells suggest that either the survival of platelets and mature myeloid cells differs or megakaryocytopoeisis is abnormal within the GPI-deficient clone. C1 NHLBI,HEMATOL BRANCH,BETHESDA,MD 20892. NIH,DEPT TRANSFUS MED,WARREN GRANT MAGNUSON CLIN CTR,BETHESDA,MD 20892. NR 22 TC 21 Z9 23 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD, ENGLAND OX5 1GB SN 0049-3848 J9 THROMB RES JI Thromb. Res. PD SEP 15 PY 1996 VL 83 IS 6 BP 433 EP 447 DI 10.1016/0049-3848(96)00153-3 PG 15 WC Hematology; Peripheral Vascular Disease SC Hematology; Cardiovascular System & Cardiology GA VH631 UT WOS:A1996VH63100003 PM 8885138 ER PT J AU Shimizu, YK Igarashi, H Kiyohara, T Cabezon, T Farci, P Purcell, RH Yoshikura, H AF Shimizu, YK Igarashi, H Kiyohara, T Cabezon, T Farci, P Purcell, RH Yoshikura, H TI A hyperimmune serum against a synthetic peptide corresponding to the hypervariable region 1 of hepatitis C virus can prevent viral infection in cell cultures SO VIROLOGY LA English DT Article ID IN-VITRO; NEUTRALIZATION; CHIMPANZEES; ANTIBODIES; ENVELOPE AB To investigate whether a principal neutralization epitope exists in hypervariable region 1 (HVR1) within the putative envelope of hepatitis C virus (HCV), we generated a hyperimmune rabbit serum against a synthetic peptide corresponding to HVR1 of HCV isolate H77. The reactivity of the serum in the enzyme-linked immunosorbent assay was correlated with the 13 amino acids (position 398-410) in HVR1. The serum prevented infection with H77 virus in cell cultures but did not prevent infection with H90 virus, a genetically divergent isolate from the same patient. The study demonstrated that neutralization of HCV was mediated, in part, by isolate-specific antibody recognizing HVR1. (C) 1996 Academic Press, Inc. C1 UNIV TOKYO,FAC MED,DEPT HEPATITIS VIRUSES SKBB,BUNKYO KU,TOKYO 113,JAPAN. NIH,DEPT VIRAL DIS & VACCINE CONTROL,TOKYO 208,JAPAN. SMITHKLINE BEECHAM BIOL,RIXERSANT,BELGIUM. NIH,DEPT INFECT DIS,BETHESDA,MD 20892. RP Shimizu, YK (reprint author), UNIV TOKYO,FAC MED,DEPT BACTERIOL,BUNKYO KU,7-3-1 HONGO,TOKYO 113,JAPAN. RI Cheng, Yushao/E-6256-2011 NR 15 TC 147 Z9 155 U1 0 U2 4 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0042-6822 J9 VIROLOGY JI Virology PD SEP 15 PY 1996 VL 223 IS 2 BP 409 EP 412 DI 10.1006/viro.1996.0497 PG 4 WC Virology SC Virology GA VJ810 UT WOS:A1996VJ81000020 PM 8806581 ER PT J AU Smanik, PA Liu, Q Furminger, TL Ryu, K Xing, S Mazzaferri, EL Jhiang, SM AF Smanik, PA Liu, Q Furminger, TL Ryu, K Xing, S Mazzaferri, EL Jhiang, SM TI Cloning of the human sodium iodide symporter SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article AB The iodide concentrating activity of the thyroid gland is essential to the production of thyroid hormone and also provides a mechanism for the treatment of thyroid cancer by radioiodine ablation. We report here the nucleotide and amino acid sequence of the human sodium iodide symporter (hNIS), which mediates the iodide uptake activity in the thyroid gland. An open reading frame of 1929 nucleotides encodes a protein of 643 amino acids with 84% identity to the rat NIS (rNIS). Transient expression of the hNIS cDNA conferred perchlorate-sensitive iodide uptake to a nonthyroid cell line, COS-7. The expression of hNIS was detected at variable levels in papillary thyroid carcinoma tissues but not in any of the thyroid carcinoma cell lines that have lost the iodide uptake activity. (C) 1996 Academic Press, Inc. C1 OHIO STATE UNIV,DEPT PHYSIOL,COLUMBUS,OH 43210. OHIO STATE UNIV,DEPT INTERNAL MED,COLUMBUS,OH 43210. NATL INST DRUG ABUSE,NIH,MOL NEUROBIOL LAB,BALTIMORE,MD 21224. RI Liu, Qing-Rong/A-3059-2012; Ain, Kenneth/A-5179-2012 OI Liu, Qing-Rong/0000-0001-8477-6452; Ain, Kenneth/0000-0002-2668-934X FU NCI NIH HHS [RZ9 CA60074] NR 7 TC 373 Z9 396 U1 0 U2 10 PU ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD SEP 13 PY 1996 VL 226 IS 2 BP 339 EP 345 DI 10.1006/bbrc.1996.1358 PG 7 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA VJ525 UT WOS:A1996VJ52500008 PM 8806637 ER PT J AU Merrick, BA He, CY Witcher, LL Patterson, RM Reid, JAJ PencePawlowski, PM Selkirk, JK AF Merrick, BA He, CY Witcher, LL Patterson, RM Reid, JAJ PencePawlowski, PM Selkirk, JK TI HSP binding and mitochondrial localization of p53 protein in human HT1080 and mouse C3H10T1/2 cell lines SO BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY LA English DT Article DE chaperone; p53 protein; heat shock protein; binding; mitochondrion; (human); (mouse) ID HEAT-SHOCK PROTEIN; TUMOR-SUPPRESSOR PROTEIN; ACTIVATING MUTATIONS; PEPTIDE; FAMILY; EXPRESSION; COMPLEXES; GENES; PHENOTYPE; ISOFORMS AB In normal cells, the tumor suppressor actions of p53 protein are mediated by specific DNA binding and protein-protein interactions within the nucleus. Mutant p53 proteins, however, often assume an aberrant conformation devoid of tumor suppressor activity and newly capable of binding to the cognate or inducible HSP70. Recent reports from our laboratory and others show that additional unknown proteins may also complex with mutant p53. In this study, we characterize p53:HSP complexes and their subcellular location in the transformed cell lines, human HT1080 and murine C3H10T1/2, which both contain aberrant p53 conformers. Immunoprecipitation and SDS-PAGE of p53 from whole cell lysates revealed the additional presence of a broad 70 kDa band and a 90 kDa band in both lines, while p53 isolated from nuclear lysates was free from other proteins. 2D-PAGE was used to isolate and identify HSP members from cytoplasmic and nuclear lysates by immunoprecipitation, Western blotting and protein sequencing. Anti-p53 immune complexes from cytoplasmic lysates contained not only HSC70 but also GRP75, GRP78 and a weakly basic 90 kDa protein, which may be related to HSP90. The inducible form of HSP70 was not complexed to p53 protein, even though expressed in these cells. Analysis of anti-HSP70, anti-GRP75 and anti-HSP90 immune complexes suggests that HSP members exist as preformed complexes in the cytoplasm, but not the nucleus. The presence of the mitochondrial and endoplasmic reticular chaperones, GRP75 and GRP78. in p53:HSP complexes suggested that p53 might be found in these cytoplasmic organelles which was confirmed in mitochondria by biochemical and immunoelectron microscopic evidence. These studies suggest that newly identified members of p53:HSP complexes represent components of a chaperone program which affects the subcellular distribution of p53 protein in these transformed lines. C1 NIEHS,LAB EXPT PATHOL,RES TRIANGLE PK,NC 27709. RP Merrick, BA (reprint author), NIEHS,MOL CARCINOGENESIS LAB,POB 12233,RES TRIANGLE PK,NC 27709, USA. NR 43 TC 41 Z9 42 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0167-4838 J9 BBA-PROTEIN STRUCT M JI Biochim. Biophys. Acta-Protein Struct. Molec. Enzym. PD SEP 13 PY 1996 VL 1297 IS 1 BP 57 EP 68 PG 12 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA VH426 UT WOS:A1996VH42600008 PM 8841381 ER PT J AU Rowles, J Scherer, SW Xi, T Majer, M Nickle, DC Rommens, JM Popov, KM Harris, RA Riebow, NL Xia, J Tsui, LC Bogardus, C Prochazka, M AF Rowles, J Scherer, SW Xi, T Majer, M Nickle, DC Rommens, JM Popov, KM Harris, RA Riebow, NL Xia, J Tsui, LC Bogardus, C Prochazka, M TI Cloning and characterization of PDK4 on 7q21.3 encoding a fourth pyruvate dehydrogenase kinase isoenzyme in human SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID DEPENDENT DIABETES-MELLITUS; PIMA-INDIANS; INSULIN-RESISTANCE; MOLECULAR-CLONING; PROTEIN-KINASES; LINKAGE; GENE; EXPRESSION; MARKERS; ASSOCIATION AB Different isoenzymes of pyruvate dehydrogenase kinase (PDK) inhibit the mitochondrial pyruvate dehydrogenase complex by phosphorylation of the E1 alpha subunit, thus contributing to the regulation of glucose metabolism. By positional cloning in the 7q21.3-q22.1 region linked with insulin resistance and non-insulin-dependent diabetes mellitus in the Pima Indians, Fee identified a gene encoding an additional human PDR isoform, as evidenced by its amino acid sequence identity (>65%) with other mammalian PDKs, and confirmed by biochemical analyses of the recombinant protein. We performed detailed comparative analyses of the gene, termed PDK4, in insulin-resistant and insulin-sensitive Pima Indians, and detected five DNA variants with comparable frequencies in both subject groups. Using quantitative reverse transcription polymerase chain reaction, we found that the variants identified in the promoter and 5'-untranslated region did not correlate with differences in mRNA level in skeletal muscle and adipose tissue. We conclude that alterations in PDK4 are unlikely to be the molecular basis underlying the observed linkage at 7q21.3-q22.1 in the Pima Indians. Information about the genomic organization and promoter sequences of PDK4 will be useful in studies of other members of this family of mitochondrial protein kinases that are important for the regulation of glucose metabolism. C1 NIDDK,CLIN DIABET & NUTR SECT,PHOENIX EPIDEMIOL & CLIN RES BRANCH,NIH,PHOENIX,AZ 85016. HOSP SICK CHILDREN,DEPT GENET,TORONTO,ON M5G 1X8,CANADA. INDIANA UNIV,SCH MED,DEPT BIOCHEM & MOL BIOL,INDIANAPOLIS,IN 46202. RI Tsui, Lap-chee/A-1081-2010; Howe, Jennifer/I-9013-2012; Scherer, Stephen /B-3785-2013 OI Scherer, Stephen /0000-0002-8326-1999 FU NIDDK NIH HHS [DK 47844]; NIGMS NIH HHS [GM 51262] NR 32 TC 130 Z9 141 U1 0 U2 3 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD SEP 13 PY 1996 VL 271 IS 37 BP 22376 EP 22382 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VG672 UT WOS:A1996VG67200018 PM 8798399 ER PT J AU Sheu, FS Mahoney, CW Seki, K Huang, KP AF Sheu, FS Mahoney, CW Seki, K Huang, KP TI Nitric oxide modification of rat brain neurogranin affects its phosphorylation by protein kinase C and affinity for calmodulin SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID LONG-TERM POTENTIATION; BINDING DOMAIN; SUBSTRATE RC3; SUBCELLULAR-FRACTIONS; DEPENDENT TARGET; NMDA RECEPTORS; NEUROMODULIN; IDENTIFICATION; SITE; PEROXYNITRITE AB Neurogranin (Ng) is a prominent protein kinase C (PKC) substrate which binds calmodulin (CaM) in the absence of Ca2+. Rat brain Ng contains four cysteine residues that were readily oxidized by nitric oxide (NO) donors, 1,1-diethyl-2-hydroxy-2-nitrosohydrazine (DEANO) and sodium nitroprusside, and by oxidants, H2O2 and o-iodosobenzoic acid, NO oxidation of Ng resulted in a conformational change detectable by increased electrophoretic mobility upon SDS-polyacrylamide gel electrophoresis. The NO-mediated mobility shift was reversed by treatment with dithiothreitol and was blocked by modification of Ng sulfhydryl groups with 4-vinylpyridine. Both the nonphosphorylated and PKC-phosphorylated Ng were susceptible to NO oxidation. Modification of Ng by DEANO was blocked by CaM in the absence of Ca2+; while in the presence of Ca2+, CaM did not protect Ng from oxidation by DEANO. CaM also failed to protect DEANO-mediated oxidation of PKC-phosphorylated Ng with or without Ca2+. Oxidation of Ng by the various oxidants apparently resulted in the formation of intramolecular disulfide bond(s) as judged by a reduction of apparent M(r) on SDS-polyacrylamide gel electrophoresis; this oxidized form, unlike the reduced form, did not bind to CaM-affinity column. The oxidized Ng was also a poorer substrate for PKC; both the reduced and oxidized forms had similar K-m values, but the V-max of the oxidized form was about one fourth of the reduced one. When comparing the rate of DEANO-mediated nitrosation of Ng with other sulfhydryl-containing compounds, it became evident that Ng ranked as one of the best NO accepters among those tested, including serum albumin, glutathione, and dithiothreitol. Ng present in the rat brain synaptosomal preparations was also oxidized by DEANO in a dose-dependent manner when analyzed by immunoblot with a polyclonal antibody against this protein. These results suggest that Ng is a likely target of NO and other oxidants and that oxidation/reduction may serve as a mechanism for controlling both the PKC phosphorylation and the CaM-binding affinity of this protein. C1 NICHHD, METAB REGULAT SECT, ENDOCRINOL & REPROD RES BRANCH, NIH, BETHESDA, MD 20892 USA. RI SHEU, Fwu-Shan/G-6435-2011 OI SHEU, Fwu-Shan/0000-0002-2784-2529 NR 50 TC 43 Z9 43 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 EI 1083-351X J9 J BIOL CHEM JI J. Biol. Chem. PD SEP 13 PY 1996 VL 271 IS 37 BP 22407 EP 22413 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VG672 UT WOS:A1996VG67200022 PM 8798403 ER PT J AU Ruocco, MR Chen, XE Ambrosino, C Dragonetti, E Liu, WM Mallardo, M DeFalco, G Palmieri, C Franzoso, G Quinto, I Venuta, S Scala, G AF Ruocco, MR Chen, XE Ambrosino, C Dragonetti, E Liu, WM Mallardo, M DeFalco, G Palmieri, C Franzoso, G Quinto, I Venuta, S Scala, G TI Regulation of HIV-1 long terminal repeats by interaction of C/EBP(NF-IL6) and NF-kappa B/Rel transcription factors SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; LOOP-HELIX PROTEINS; C/EBP FAMILY; DNA-BINDING; INTERLEUKIN-8 GENE; NUCLEAR FACTOR; P50 SUBUNIT; EXPRESSION; TYPE-1; INFECTION AB We report the characterization of a CAAT enhancer-binding protein (C/EBP) (NF-IL6) element encompassing the region from -174 to -166 of the U3 long terminal repeat (LTR) region of HIV-1. This C/EBP cis sequence was found to bind to C/EBP beta and C/EBP delta factors in DNA band shift assay. Transfection of NTera-2 cells with a HIV-1-LTR CAT construct (pC15CAT), together with C/EBP beta or C/EBP delta expression plasmids showed that C/EBP proteins strongly activated the HIV-1 promoter. Deletions encompassing the C/EBP-binding site resulted in the enhancement of the LTR activation mediated by C/EBP proteins, suggesting that other sequences located 3' to -170 were indeed the target for C/EBP factors, This possibility was confirmed by using the pCD54E9CAT plasmid, in which the NF-kappa B enhancer was inserted 5' to the HIV-1 LTR TATA box. A NF-kappa B1(p50) expression plasmid was also utilized to test for functional co-operation between NF-kappa B and C/EBP factors. We observed that p50 . C/EBP beta and p50 . C/EBP delta complexes were generated in tested cells and strongly activated the HIV-1 LTR by binding to the NF-kappa B sequences, The physical association of NF-kappa B1(p50) with C/EBP factors was assayed by direct interaction of in vitro translated p50 proteins with C/EBP beta or C/EBP delta produced as glutathione S-transferase fusion proteins. Moreover, p50 . C/EBP beta complexes were observed in vivo by using DNA affinity studies with biotinylated NF-kappa B oligonucleotides. By using mutant forms of p50 or C/EBP beta proteins we found that the transactivation of HIV-1 LTR by p50 . C/EBP beta complexes required the DNA-binding domain of p50 and the transcription activation domain of C/EBP beta. C1 UNIV REGGIO CALABRIA,SCH MED,INST CLIN & EXPT MED,I-88100 CATANZARO,ITALY. UNIV NAPLES FEDERICO II,SCH MED,DEPT BIOCHEM & BIOMED TECHNOL,I-80131 NAPLES,ITALY. NIAID,IMMUNOREGULAT LAB,NIH,BETHESDA,MD 20892. RI SCALA, GIUSEPPE/A-3280-2009; Palmieri, Camillo/A-3195-2009 OI Palmieri, Camillo/0000-0001-6707-4723 NR 63 TC 85 Z9 85 U1 0 U2 4 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD SEP 13 PY 1996 VL 271 IS 37 BP 22479 EP 22486 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VG672 UT WOS:A1996VG67200032 PM 8798413 ER PT J AU Hartman, JL Northup, JK AF Hartman, JL Northup, JK TI Functional reconstitution in situ of 5-hydroxytryptamine(2c) (5HT(2c)) receptors with alpha(q) and inverse agonism of 5HT(2c) receptor antagonists SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID PROTEIN-COUPLED RECEPTORS; PHOSPHOLIPASE-C ISOZYMES; BETA-GAMMA-SUBUNITS; GTP-BINDING PROTEIN; SF9 CELLS; SIGNAL-TRANSDUCTION; GUANINE-NUCLEOTIDES; INTRINSIC ACTIVITY; ADENYLATE-CYCLASE; CHOROID-PLEXUS AB Membranes prepared after infection of Sf9 cells with recombinant baculovirus containing the rat 5HT(2c) receptor DNA, but not after infection with wild-type virus, expressed high affinity binding sites for I-125-lysergic acid diethylamide and [H-3]mesulergine. The receptor site density reached an optimum of 50-70 pmol/mg membrane protein at 60 h postinfection. Extraction of peripheral membrane proteins from the postnuclear membrane fraction with 6 M urea depleted GTP gamma S-binding 4-fold without decreasing 5HT(2c) receptor binding activity. Urea-extracted Sf9 membranes expressing the 5HT(2c) receptor catalyzed the activation of squid retinal alpha(q) but not bovine retinal alpha(t) or bovine alpha(o) alpha(i). Productive interaction of 5HT(2c), receptors with squid alpha(q) was enhanced by the addition of beta gamma dimers prepared from either bovine brain or bovine rod outer segment discs. While the addition of serotonin increased 5HT(2c) receptor-catalyzed GTP gamma S binding to alpha(q), the unoccupied receptor was also catalytically active. The 5HT(2c) receptor antagonists, mesulergine, mianserin, and ketanserin competitively inhibited 5HT activation of the receptor with predicted rank-order affinities; and mianserin and ketanserin markedly inhibited basal 5HT(2c) receptor activity. Interestingly, this ''inverse agonist'' efficacy did not correlate with antagonist affinity for the 5HT(2c) receptor. Baculoviral expression of the 5HT(2c) receptor and urea extraction of postnuclear Sf9 cell membranes have provided a high density of in situ, uncoupled, G-protein-linked receptor useful for reconstitution with purified G-protein subunits. This has allowed for independent manipulation of receptor and G protein chemical concentrations and has revealed that a G-protein-linked receptor can possess a significant basal catalytic activity and that antagonist compounds can act as inverse agonists of this basal activity at the level of receptor activation of G-proteins. RP NIMH, CELL BIOL LAB, BLDG 36, BETHESDA, MD 20892 USA. NR 47 TC 75 Z9 75 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 EI 1083-351X J9 J BIOL CHEM JI J. Biol. Chem. PD SEP 13 PY 1996 VL 271 IS 37 BP 22591 EP 22597 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VG672 UT WOS:A1996VG67200047 PM 8798428 ER PT J AU Matsumoto, K Meric, F Wolffe, AP AF Matsumoto, K Meric, F Wolffe, AP TI Translational repression dependent on the interaction of the Xenopus Y-box protein FRGY2 with mRNA - Role of the cold shock domain, tail domain, and selective RNA sequence recognition SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID MESSENGER-RIBONUCLEOPROTEIN PARTICLES; OOCYTE-SPECIFIC PROTEINS; ACID BINDING-PROTEINS; DEVELOPMENTAL REGULATION; TRANSCRIPTION FACTOR; BACILLUS-SUBTILIS; MASKING PROTEINS; SOMATIC-CELLS; DNA-BINDING; IN-VIVO AB We have examined the determinants of the translational repression of mRNA by the Xenopus oocyte-specific Y-box protein FRGY2 using in vitro and in vivo assays. In vitro reconstitution of messenger ribonucleoprotein (mRNP) complexes demonstrates that the sequence-specific RNA-binding cold shock domain is not required for translational repression, whereas the RNA-binding C-terminal tail domain is essential. However, microinjection of reconstituted mRNPs into Xenopus oocytes demonstrates that although translational repression occurs in the absence of consensus RNA binding sequences for FRGY2, the presence of FRGY2 recognition elements within mRNA potentiates translational repression. Analysis of the in vivo assembly of mRNP shows that the cold shock domain alone is not stably incorporated into mRNP, whereas the C-terminal tail domain is sufficient for stable incorporation. We suggest that translational repression of mRNA by FRGY2 is favored by sequence-selective recognition of RNA sequences by the cold shock domain. However, translational repression in vitro and the assembly of mRNP in vivo requires the relatively nonspecific interaction of the C-terminal tail domain with mRNA. Thus two distinct domains of FRGY2 are likely to contribute to translational control. C1 NICHHD,MOL EMBRYOL LAB,NIH,BETHESDA,MD 20892. RI Matsumoto, Ken/F-9083-2013 OI Matsumoto, Ken/0000-0002-7864-3394 NR 43 TC 99 Z9 100 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD SEP 13 PY 1996 VL 271 IS 37 BP 22706 EP 22712 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VG672 UT WOS:A1996VG67200063 PM 8798444 ER PT J AU Wang, HS Cao, HJ Winn, VD Rezanka, LJ Frobert, Y Evans, CH Sciaky, D Young, DA Smith, TJ AF Wang, HS Cao, HJ Winn, VD Rezanka, LJ Frobert, Y Evans, CH Sciaky, D Young, DA Smith, TJ TI Leukoregulin induction of prostaglandin-endoperoxide H synthase-2 in human orbital fibroblasts - An in vitro model for connective tissue inflammation SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HUMAN DERMAL FIBROBLASTS; HUMAN-SKIN FIBROBLASTS; NF-KAPPA-B; CYCLIC ADENOSINE-MONOPHOSPHATE; VESICULAR GLAND MICROSOMES; CULTURED HUMAN-FIBROBLASTS; DENOVO PROTEIN-SYNTHESIS; CELL-DERIVED CYTOKINE; GRAVES OPHTHALMOPATHY; INTERFERON-GAMMA AB Several proinflammatory cytokines can increase prostaglandin E(2) (PGE(2)) synthesis in a variety of cell types, constituting an important component of the inflammatory response. We demonstrate here that leukoregulin, a 50-kDa product of activated T lymphocytes, dramatically increases PGE, synthesis in cultured human orbital fibroblasts. This up-regulation is mediated through an induction of prostaglandin-endoperoxide H synthase-2 (PGHS-2), the inflammatory cyclooxygenase. Steady-state levels of PGHS-2 mRNA are increased within 1.5 h of leukoregulin addition and are near maximal. by 6 h, when they are 50-fold or higher above basal levels, The increase in PGHS-2 mRNA levels is partially blocked by cycloheximide, suggesting de novo synthesis of an intermediate protein may be required for a maximal leukoregulin response. Nuclear run-on studies indicate PGHS-2 gene transcription is up-regulated by leukoregulin a-fold after 2 and 6 h. PGHS-2 protein, as assessed by Western blotting and two-dimensional protein gel analysis, is increased dramatically in orbital fibroblasts, This lymphokine-dependent expression of PGHS-2 is blocked by dexamethasone, and the increase in PGE, and cAMP levels following leukoregulin treatment is also blocked by indomethacin and by SC 58125, a newly developed PGHS-2-selective cyclooxygenase inhibitor. The dramatic increase in cAMP levels causes marked alteration in orbital fibroblast morphology. PGHS-2 expression in dermal fibroblasts is also increased by leukoregulin; however, the response is considerably less robust, and these cells do not undergo a change in morphology. Both orbital and dermal fibroblasts express high levels of PGHS-1 mRNA and protein, the other abundant form of cyclooxygenase. In contrast to its effects on PGHS-2 expression, leukoregulin fails to alter PGHS-1 levels in either orbital or dermal fibroblasts, suggesting that PGHS-1 is not involved in cytokine-dependent prostanoid production in human fibroblasts. The increased PGHS-2 expression elicited by leukoregulin in orbital fibroblasts may be a consequence of both transcriptional and post-transcriptional effects. These observations help clarify the pathogenic mechanism relevant to the intense inflammation associated with Graves' ophthalmopathy. Lymphocytes trafficked to orbital tissues have a putative role, through the cytokines they release, in the activation of fibroblasts in this autoimmune disease. C1 ALBANY MED COLL,DEPT MED,DIV MOL & CELLULAR MED,ALBANY,NY 12208. ALBANY MED COLL,DEPT BIOCHEM & MOL BIOL,ALBANY,NY 12208. SAMUEL S STRATTON VET AFFAIRS MED CTR,ALBANY,NY 12208. UNIV ROCHESTER,SCH MED & DENT,DEPT MED,DIV ENDOCRINOL & METAB,E HENRY KEUTMANN LABS,ROCHESTER,NY 14642. UNIV ROCHESTER,SCH MED & DENT,DEPT BIOCHEM,ROCHESTER,NY 14642. NCI,BIOL LAB,BETHESDA,MD 20892. CEA SACLAY,SERV PHARMACOL & IMMUNOL,F-91191 GIF SUR YVETTE,FRANCE. OI Smith, Terry/0000-0002-6279-9685 FU NEI NIH HHS [EY 08976]; NIDDK NIH HHS [DK 16177] NR 81 TC 75 Z9 78 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD SEP 13 PY 1996 VL 271 IS 37 BP 22718 EP 22728 PG 11 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA VG672 UT WOS:A1996VG67200065 PM 8798446 ER EF