FN Thomson Reuters Web of Science™ VR 1.0 PT J AU Rando, OJ Zhao, KJ Crabtree, GR AF Rando, OJ Zhao, KJ Crabtree, GR TI Searching for a function for nuclear actin SO TRENDS IN CELL BIOLOGY LA English DT Review ID ABL TYROSINE KINASE; BINDING-PROTEIN; DEPOLYMERIZING PROTEIN; DEOXYRIBONUCLEASE-I; MATRIX PREPARATIONS; MESSENGER-RNA; F-ACTIN; CELLS; COFILIN; LOCALIZATION AB The abundant cytoskeletal protein actin was numerous cytoplasmic roles. Although there are many reports of the presence of actin in the nucleus, in general they have been discounted as artifactual. However, recent work has begun to provide evidence far important roles for actin in nuclear processes ranging from chromatin remodelling to splicing. In addition, several regulators of actin polymerization are localized to the nucleus or translocate to the nucleus in a regulated manner, suggesting that there is some function of actin in the nucleus that is subject to regulation. This review discusses the evidence for actin in the nucleus and summarizes recent work suggesting that actin or actin-related proteins are involved in the regulation of nuclear processes such as chromatin remodelling. C1 Stanford Univ, Sch Med, Stanford, CA 94305 USA. NHLBI, Lab Mol Immunol, NIH, Bethesda, MD 20892 USA. RP Rando, OJ (reprint author), Stanford Univ, Sch Med, Stanford, CA 94305 USA. NR 57 TC 207 Z9 216 U1 0 U2 5 PU ELSEVIER SCIENCE LONDON PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0962-8924 J9 TRENDS CELL BIOL JI Trends Cell Biol. PD MAR PY 2000 VL 10 IS 3 BP 92 EP 97 DI 10.1016/S0962-8924(99)01713-4 PG 6 WC Cell Biology SC Cell Biology GA 286AH UT WOS:000085420200004 PM 10675902 ER PT J AU Strausberg, RL Buetow, KH Emmert-Buck, MR Klausner, RD AF Strausberg, RL Buetow, KH Emmert-Buck, MR Klausner, RD TI The cancer genome anatomy project - Building an annotated gene index SO TRENDS IN GENETICS LA English DT Article ID DISCOVERY; CATALOG C1 NCI, Bethesda, MD 20892 USA. RP Strausberg, RL (reprint author), NCI, Bethesda, MD 20892 USA. NR 13 TC 80 Z9 83 U1 0 U2 1 PU ELSEVIER SCIENCE LONDON PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0168-9525 J9 TRENDS GENET JI Trends Genet. PD MAR PY 2000 VL 16 IS 3 BP 103 EP 106 DI 10.1016/S0168-9525(99)01937-X PG 4 WC Genetics & Heredity SC Genetics & Heredity GA 288NF UT WOS:000085568600002 PM 10689348 ER PT J AU Zhang, JZ AF Zhang, JZ TI Protein-length distributions for the three domains of life SO TRENDS IN GENETICS LA English DT Editorial Material ID DIVERGENCE; ARCHAEA C1 NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. Penn State Univ, Inst Mol Evolutionary Genet, University Pk, PA 16802 USA. RP Zhang, JZ (reprint author), NIAID, Host Def Lab, NIH, Bldg 10,Room 11N104,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 11 TC 86 Z9 90 U1 0 U2 7 PU ELSEVIER SCIENCE LONDON PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0168-9525 J9 TRENDS GENET JI Trends Genet. PD MAR PY 2000 VL 16 IS 3 BP 107 EP 109 DI 10.1016/S0168-9525(99)01922-8 PG 3 WC Genetics & Heredity SC Genetics & Heredity GA 288NF UT WOS:000085568600003 PM 10689349 ER PT J AU Belnap, DM Steven, AC AF Belnap, DM Steven, AC TI 'Deja vu all over again': the similar structures of bacteriophage PRD1 and adenovirus SO TRENDS IN MICROBIOLOGY LA English DT Review ID CRYSTAL-STRUCTURE; PROTEIN; RESOLUTION; TOPOLOGY C1 NIAMSD, Struct Biol Res Lab, NIH, Bethesda, MD 20892 USA. RP Belnap, DM (reprint author), NIAMSD, Struct Biol Res Lab, NIH, Bethesda, MD 20892 USA. NR 15 TC 27 Z9 28 U1 0 U2 5 PU ELSEVIER SCIENCE LONDON PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0966-842X J9 TRENDS MICROBIOL JI Trends Microbiol. PD MAR PY 2000 VL 8 IS 3 BP 91 EP 93 DI 10.1016/S0966-842X(00)01704-2 PG 3 WC Biochemistry & Molecular Biology; Microbiology SC Biochemistry & Molecular Biology; Microbiology GA 292JB UT WOS:000085789800002 PM 10707053 ER PT J AU Sanders, JM Chen, LJ Burka, LT Matthews, HB AF Sanders, JM Chen, LJ Burka, LT Matthews, HB TI Metabolism and disposition of luminol in the rat SO XENOBIOTICA LA English DT Article ID CHEMILUMINESCENCE AB 1. The metabolism and disposition of Luminol (LMN. 3-aminophthalhydrazide), a widely used forensic and laboratory reagent that chemiluminesses upon oxidation, was determined as part of its overall toxicological characterization. 2. Radiolabelled LMN was well absorbed, metabolized and excreted following p.o. administration of a range of doses. About 90 % of the total dose was recovered within 21 h of administration in urine in the form of two metabolites identified as LMN N-8-glucuronide and LMN N-8-sulphamic acid. 3-Aminophthalic acid, the oxidative product of LMN in the light-emitting reaction, was apparently not formed in vivo. 3. Metabolism and disposition of an i.v. administered dose was similar to that following gavage. Little or no LMN-derived radioactivity was present in tissue within 12 h postdosing. Excretion of radioactivity in bile following i.v. injection was minimal (similar to 8 % of the total dose in 6 h) and consisted of the same urinary-excreted glucuronide and sulphate conjugates. 4. LMN was not absorbed dermally in rat, potentially a major route of exposure to human. If the fate of LMN is similar between species, this compound should have little potential for either dermal absorption, bioaccumulation in tissues following other routes of exposure or chronic toxicity in humans. C1 NIEHS, Natl Toxicol Program, Lab Pharmacol & Chem, Res Triangle Pk, NC 27709 USA. RP Sanders, JM (reprint author), NIEHS, Natl Toxicol Program, Lab Pharmacol & Chem, POB 12233, Res Triangle Pk, NC 27709 USA. NR 20 TC 14 Z9 14 U1 0 U2 1 PU TAYLOR & FRANCIS LTD PI LONDON PA 11 NEW FETTER LANE, LONDON EC4P 4EE, ENGLAND SN 0049-8254 J9 XENOBIOTICA JI Xenobiotica PD MAR PY 2000 VL 30 IS 3 BP 263 EP 272 PG 10 WC Pharmacology & Pharmacy; Toxicology SC Pharmacology & Pharmacy; Toxicology GA 294DN UT WOS:000085896200004 PM 10752641 ER PT J AU Nabel, EG AF Nabel, EG TI New approaches to pulmonary hypertension - Will therapies in mice work in humans? SO CIRCULATION LA English DT Editorial Material DE editorials; hypertension, pulmonary; gene therapy ID GENE-RELATED PEPTIDE; K+ CHANNELS; CALCITONIN C1 NHLBI, NIH, Bethesda, MD 20892 USA. RP Nabel, EG (reprint author), NHLBI, NIH, Bldg 10,Room 8C103, Bethesda, MD 20892 USA. NR 6 TC 3 Z9 3 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 29 PY 2000 VL 101 IS 8 BP 839 EP 840 PG 2 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 288MZ UT WOS:000085568000013 PM 10694519 ER PT J AU Carlson, BA Kwon, SY Lee, BJ Hatfield, D AF Carlson, BA Kwon, SY Lee, BJ Hatfield, D TI Yeast asparagine (Asn) tRNA without Q base promotes eukaryotic frameshifting more efficiently than mammalian Asn tRNAs with or without Q base SO MOLECULES AND CELLS LA English DT Article DE asparagine tRNA; queuosine base; ribosomal frameshifting; wheat germ extracts ID TRANSFER-RNA; TRANSLATIONAL SUPPRESSION; RIBOSOMAL FRAMESHIFT; GENE-EXPRESSION; TMV RNA; VIRUS; CODON; READTHROUGH; SEQUENCE; PROTEIN AB In this study, we compare the efficiency of Asn tRNA from mammalian sources with and without the highly modified queuosine (Q) base in the wobble position of its anticodon and Asn tRNA from yeast, which naturally lacks Q base, to promote frameshifting, Interestingly, no differences in the ability of the two mammalian Asn tRNAs to promote frameshifting were observed, while yeast tRNA(Asn)-Q promoted frameshifting more efficiently than its mammalian counterparts in both rabbit reticulocyte lysates and wheat germ extracts. The shiftability of yeast Asn tRNA is therefore not due, or at least not completely, to the lack of Q base and most likely the shiftiness resides in structural differences elsewhere in the molecule. However, we cannot absolutely rule out a role of Q base in frameshifting as wheat germ extracts and a lysate depleted of most of its tRNA and supplemented with calf liver tRNA contain both Asn tRNA with or without Q base. C1 NCI, Sect Mol Biol Selenium, Lab Basic Res, NIH, Bethesda, MD 20892 USA. NCI, Div Basic Sci, NIH, Bethesda, MD 20892 USA. Seoul Natl Univ, Inst Mol Biol & Genet, Mol Genet Lab, Seoul 151742, South Korea. RP Hatfield, D (reprint author), NCI, Sect Mol Biol Selenium, Lab Basic Res, NIH, Bethesda, MD 20892 USA. EM hatfield@dc37a.nci.nih.gov RI Kwon, So Yeon/M-5625-2014 OI Kwon, So Yeon/0000-0002-8490-9101 NR 25 TC 8 Z9 8 U1 0 U2 0 PU KOREAN SOC MOLECULAR & CELLULAR BIOLOGY PI SEOUL PA 635-4, YUCKSAM-DONG, GANGNAM-GU, SEOUL 135-703, SOUTH KOREA SN 1016-8478 EI 0219-1032 J9 MOL CELLS JI Mol. Cells PD FEB 29 PY 2000 VL 10 IS 1 BP 113 EP 118 DI 10.1007/s100590070019 PG 6 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 291DQ UT WOS:000085719200019 PM 10774757 ER PT J AU Rovescalli, AC Cinquanta, M Ferrante, J Kozak, CA Nirenberg, M AF Rovescalli, AC Cinquanta, M Ferrante, J Kozak, CA Nirenberg, M TI The mouse Nkx-1.2 homeobox gene: Alternative RNA splicing at canonical and noncanonical splice sites SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE Sax-1; transcription factors; embryo; brain; spermatozoa ID MINOR CLASS; IN-VIVO; EXPRESSION; SPERMATOZOA; DROSOPHILA; CELLS; EMBRYOGENESIS; TRANSCRIPTION; INTRONS; REGION AB A mouse homeobox gene, Nkx-1.2, (previously termed Sax-1) that is closely related to the Drosophila NK-1/S59 gene was cloned, and genomic DNA and cDNA were sequenced. Nine Nkx-1.2 cDNA clones were found that correspond to three species of Nkx-1.2 mRNA that are formed by alternative splicing at conventional 5' donor and 3' acceptor splice sites; however, seven cDNA clones were found that correspond to three species of Nkx-1.2 mRNA from testes that have novel TG/AC 5' and 3' splice sites. The consensus splice sequences are: 5' donor, CC down arrow TGGAAG; 3' acceptor, ACT-TAC down arrow. Predicted amino acid sequences suggest that some transcripts may be translated into proteins that lack part or all of the homeodomain. At least three bands of Nkx-1.2 mRNA were found in RNA from the testes. Nkx-1.2 mRNA was shown to be present in postmeiotic germ cells of the testis and in mature spermatozoa, Nkx-1.2 mRNA also was found in regions of the adult cerebral cortex, hippocampus, diencephalon, pons/medulla, and cerebellum. Nkx-1.2 mRNA was found in embryos in highest abundance in 10-day embryos; the mRNA levels decrease during further development, Nkx-1.2 mRNA also was found in discrete zones of the embryonic mesencephalon and myelencephalon. C1 NHLBI, Lab Biochem Genet, NIH, Bethesda, MD 20892 USA. NIAID, Mol Microbiol Lab, NIH, Bethesda, MD 20892 USA. RP Nirenberg, M (reprint author), NHLBI, Lab Biochem Genet, NIH, Bldg 36,Room 1C-06,36 Convent Dr,MSC 4036, Bethesda, MD 20892 USA. NR 30 TC 7 Z9 10 U1 1 U2 1 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD FEB 29 PY 2000 VL 97 IS 5 BP 1982 EP 1987 DI 10.1073/pnas.030539397 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 289QJ UT WOS:000085633200007 PM 10681422 ER PT J AU Pommier, Y Kohlhagen, G Pourquier, P Sayer, JM Kroth, H Jerina, DM AF Pommier, Y Kohlhagen, G Pourquier, P Sayer, JM Kroth, H Jerina, DM TI Benzo[a]pyrene diol epoxide adducts in DNA are potent suppressors of a normal topoisomerase I cleavage site and powerful inducers of other topoisomerase I cleavages SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID EUKARYOTIC TOPOISOMERASE; RELIGATION REACTIONS; COVALENT COMPLEXES; NUCLEOSIDE ADDUCTS; DRUG CAMPTOTHECIN; SEQUENCE; REGION; ENANTIOMERS; INHIBITORS; MECHANISM AB The catalytic intermediates of DNA topoisomerase I (top1) are cleavage complexes that can relax DNA supercoiling (intramolecular reaction) or mediate recombinations (intermolecular religation). We report here that DNA adducts formed from benzo[a]pyrene bay-region diol epoxides can markedly affect top1 activity. Four oligonucleotide 22-mers of the same sequence were synthesized, each of which contained a stereoisomerically unique benzo[a]pyrene 7,8-diol 9,10-epoxide adduct at the 2-amino group of a central 2'-deoxyguanosine residue. These four adducts correspond to either cis or trans opening at C-10 of the (+)-(7R, 8S, 9S, 10R)- or (-)-(7S, 8R, 9R, 10S)-7,8-diol 9,10-epoxides. Their solution conformations in duplex DNA (intercalated and minor-groove bound for the cis and trans opened adducts respectively) can be deduced from previous NMR studies. All four adducts completely suppress top1 cleavage activity at the alkylation site and induce the formation of new top1 cleavage complexes on both strands of the DNA 3-6 bases away from the alkylation site. The trans opened adduct from the highly carcinogenic (+)-diol epoxide is the most active in inducing top1 cleavage independently of camptothecin, demonstrating that minor groove alkylation can efficiently poison top1. We also found that this isomer of the diol epoxide induces the formation of top1-DNA complexes in mammalian cells, which suggests a possible relationship between induction of top1 cleavage complexes and carcinogenic activity of benzo[a]pyrene diol epoxides. C1 NCI, Mol Pharmacol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. NIDDKD, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. RP Pommier, Y (reprint author), NCI, Mol Pharmacol Lab, Div Basic Sci, NIH, Bldg 37,Room 5D02, Bethesda, MD 20892 USA. NR 50 TC 60 Z9 63 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 FEB 29 PY 2000 VL 97 IS 5 BP 2040 EP 2045 DI 10.1073/pnas.040397497 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 289QJ UT WOS:000085633200017 PM 10688881 ER PT J AU Mertz, EL Krishtalik, LI AF Mertz, EL Krishtalik, LI TI Low dielectric response in enzyme active site SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID PHOTOSYNTHETIC REACTION CENTER; SOLVENT REORGANIZATION ENERGY; ELECTRON-TRANSFER; MOLECULAR-DYNAMICS; CYTOCHROME-C; BACTERIAL PHOTOSYNTHESIS; RESOLVED FLUORESCENCE; ALPHA-CHYMOTRYPSIN; PROTEIN HYDRATION; SOLID-STATE AB The kinetics of charge transfer depend crucially on the dielectric reorganization of the medium. In enzymatic reactions that involve charge transfer, atomic dielectric response of the active site and of its surroundings determines the efficiency of the protein as a catalyst. We report direct spectroscopic measurements of the reorganization energy associated with the dielectric response in the active site of alpha-chymotrypsin. A chromophoric inhibitor of the enzyme is used as a spectroscopic probe. We find that water strongly affects the dielectric reorganization in the active site of the enzyme in solution. The reorganization energy of the protein matrix in the vicinity of the active site is similar to that of low-polarity solvents. Surprisingly, water exhibits an anomalously high dielectric response that cannot be described in terms of the dielectric continuum theory. As a result, sequestering the active site from the aqueous environment inside low-dielectric enzyme body dramatically reduces the dielectric reorganization. This reduction is particularly important for controlling the rate of enzymatic reactions. C1 NICHHD, Lab Phys & Struct Biol, NIH, Bethesda, MD 20892 USA. Russian Acad Sci, AN Frumkin Electrochem Inst, Moscow 117071, Russia. RP NICHHD, Lab Phys & Struct Biol, NIH, Bldg 9,Room 1E-122,9000 Rockville Pike, Bethesda, MD 20892 USA. EM emertz@helix.nih.gov NR 64 TC 87 Z9 89 U1 1 U2 8 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 FEB 29 PY 2000 VL 97 IS 5 BP 2081 EP 2086 DI 10.1073/pnas.050316997 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 289QJ UT WOS:000085633200024 PM 10681440 ER PT J AU Emmert, S Kobayashi, N Khan, SG Kraemer, KH AF Emmert, S Kobayashi, N Khan, SG Kraemer, KH TI The xeroderma pigmentosum group C gene leads to selective repair of cyclobutane pyrimidine dimers rather than 6-4 photoproducts SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID NUCLEOTIDE EXCISION-REPAIR; TRANSCRIPTION-COUPLED REPAIR; COCKAYNE-SYNDROME CELLS; DNA-REPAIR; ULTRAVIOLET-RADIATION; SKIN-CANCER; EXPRESSION; FIBROBLASTS; NUCLEASE; COMPLEX AB We investigated the contribution of the xeroderma pigmentosum group C (XPC) gene to DNA repair. We stably transfected XPC cells (XP4PA-SV-EB) with XPC cDNA and selected a partially corrected (XP4PA-SE1) and a fully corrected (XP4PA-SE2) clone. Cell survival after UVC (254 nm) exposure was low for XP4PA-SV-EB, intermediate for XP4PA-SE1, and normal for XP4PA-SE2 cells. XP4PA-SV-EB cells had undetectable XPC mRNA and protein levels. XP4PA-SE1 cells had 130% of normal mRNA but 25% of normal protein levels, whereas XP4PA-SE2 cells had an 18-fold mRNA overexpression and normal XPC protein levels compared with normal cells. We measured cyclobutane pyrimidine dimers (CPD) and 6-4 photoproducts (6-4PP) by using specific mAbs and the ELISA technique. XP4PA-SV-EB cells had no detectable removal of CPD or 6-4PP from their global genome by 24 h after 30 J/m(2) UVC exposure. The partially corrected XP4PA-SE1 cells had normal repair of CPD but minimal repa ir of 6-4PP by 24 h, whereas the fully corrected XP4PA-SE2 cells regained normal CPD and 6-4PP repair capacities. We also exposed pRSVcat plasmid to UVC (to induce CPD and 6-4PP), to UVC + photolyase (to leave only 6-4PP on the plasmid), or to UVB + acetophenone (to induce only CPD). Host cell reactivation of UVB + acetophenone-, but not of UVC + photolyase-treated plasmids was normal in XP4PA-SE1 cells. Thus, increasing XPC gene expression leads to selective repair of CPD in the global genome. Undetectable XPC protein is associated with no repair of CPD or 6-4PP, detectable but subnormal XPC protein levels reconstitute CPD but not 6-4PP repair, and normal XPC protein levels fully reconstitute both CPD and 6-4PP repair. C1 NCI, Mol Carcinogenesis Lab, Bethesda, MD 20892 USA. NCI, Cell Biol Lab, Bethesda, MD 20892 USA. RP Kraemer, KH (reprint author), NCI, Mol Carcinogenesis Lab, Bldg 37, Bethesda, MD 20892 USA. FU Intramural NIH HHS [Z01 BC004517-31] NR 49 TC 55 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 FEB 29 PY 2000 VL 97 IS 5 BP 2151 EP 2156 DI 10.1073/pnas.040559697 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 289QJ UT WOS:000085633200036 PM 10681431 ER PT J AU Factor, VM Laskowska, D Jensen, MR Woitach, JT Popescu, NC Thorgeirsson, SS AF Factor, VM Laskowska, D Jensen, MR Woitach, JT Popescu, NC Thorgeirsson, SS TI Vitamin E reduces chromosomal damage and inhibits hepatic tumor formation in a transgenic mouse model SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID GROWTH-FACTOR-ALPHA; PROTEIN-KINASE-C; MICE; LIVER; MYC; TOCOPHEROL; CANCER; HEPATOCARCINOGENESIS; PHOSPHORYLATION; DNA AB We have previously shown that chronic activation of mitogenic signaling induced by over-expression of c-myc and transforming growth factor-alpha (TGF alpha) transgenes in mouse liver induces a state of oxidative stress. We therefore proposed that increased reactive oxygen species (ROS) generation might be responsible for the extensive chromosomal damage and acceleration of hepatocarcinogenesis characteristic: for TGF alpha/c-myc mice. In this study, we show that vitamin E (VE), a potent free radical scavenging antioxidant, is able to protect liver tissue against oxidative stress and suppress tumorigenic potential of c-myc oncogene, Dietary supplementation with VE, starting from weaning, decreased ROS generation coincident with a marked inhibition of hepatocyte proliferation while increasing the chromosomal as well as mtDNA stability in the liver. Similarly, dietary VE reduced liver dysplasia and increased viability of hepatocytes, At 6 mo of age, VE treatment decreased the incidence of adenomas by 65% and prevented malignant conversion. These results indicate that ROS generated by over-expression of c-myc and TGF alpha in the liver are the primary carcinogenic agents in this animal model. Furthermore, the data demonstrate that dietary supplementation of VE can effectively inhibit liver cancer development. C1 NCI, Expt Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. RP Thorgeirsson, SS (reprint author), NCI, Expt Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. RI Jensen, Michael/E-9677-2011 NR 31 TC 104 Z9 111 U1 0 U2 1 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD FEB 29 PY 2000 VL 97 IS 5 BP 2196 EP 2201 DI 10.1073/pnas.040428797 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 289QJ UT WOS:000085633200044 PM 10681450 ER PT J AU Kagawa, TF Cooney, JC Baker, HM McSweeney, S Liu, MY Gubba, S Musser, JM Baker, EN AF Kagawa, TF Cooney, JC Baker, HM McSweeney, S Liu, MY Gubba, S Musser, JM Baker, EN TI Crystal structure of the zymogen form of the group A Streptococcus virulence factor SpeB: An integrin-binding cysteine protease SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID PYOGENES; REFINEMENT; DISEASE; INHIBITION; EXPRESSION; ACTINIDIN; PROREGION; PROTEINS; PRODUCE; PAPAIN AB Pathogenic bacteria secrete protein toxins that weaken or disable their host, and thereby act as virulence factors. We have determined the crystal structure of streptococcal pyrogenic exotoxin B (SpeB), a cysteine protease that is a major virulence factor of the human pathogen Streptococcus pyogenes and participates in invasive disease episodes, including necrotizing fasciitis. The structure, determined for the 40-kDa precursor form of SpeB at 1.6-Angstrom, resolution, reveals that the protein is a distant homologue of the papain superfamily that includes the mammalian cathepsins B, K, L, and S. Despite negligible sequence identity, the protease portion has the canonical papain fold, albeit with major loop insertions and deletions. The catalytic site differs from most other cysteine proteases in that it lacks the Asn residue of the Cys-His-Asn triad. The prosegment has a unique fold and inactivation mechanism that involves displacement of the catalytically essential His residue by a loop inserted into the active site. The structure also reveals the surface location of an integrin-binding Arg-Gly-Asp (RGD) motif that is a feature unique to SpeB among cysteine proteases and is linked to the pathogenesis of the most invasive strains of S. pyogenes. C1 Univ Auckland, Sch Biol Sci, Auckland, New Zealand. Univ Auckland, Dept Chem, Auckland, New Zealand. Massey Univ, Inst Mol Biosci, Palmerston North, New Zealand. European Mol Biol Lab, Grenoble Outstn, F-38042 Grenoble, France. Baylor Coll Med, Dept Pathol, Inst Human Bacterial Pathogenesis, Houston, TX 77030 USA. NIAID, Rocky Mt Labs, Lab Human Bacterial Pathogenesis, NIH, Hamilton, MT 59840 USA. RP Baker, EN (reprint author), Univ Auckland, Sch Biol Sci, Private Bag 92-019, Auckland, New Zealand. FU NIAID NIH HHS [AI-33119]; PHS HHS [MR17841, MR36247] NR 46 TC 81 Z9 83 U1 1 U2 3 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 FEB 29 PY 2000 VL 97 IS 5 BP 2235 EP 2240 DI 10.1073/pnas.040549997 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 289QJ UT WOS:000085633200051 PM 10681429 ER PT J AU Liebl, DJ Klesse, LJ Tessarollo, L Wohlman, T Parada, LF AF Liebl, DJ Klesse, LJ Tessarollo, L Wohlman, T Parada, LF TI Loss of brain-derived neurotrophic factor-dependent neural crest-derived sensory neurons in neurotrophin-4 mutant mice SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID DORSAL-ROOT GANGLIA; NERVE GROWTH-FACTOR; TRK RECEPTORS; DIFFERENTIATION ANTIGENS; DISTINCT POPULATIONS; TARGETED DISRUPTION; SPINAL GANGLIA; GENE; SURVIVAL; INNERVATION AB Peripheral ganglion neurons confer sensory information including touch, pain, temperature, and proprioception. Sensory modality is linked to specific neurotrophin (NTF) requirements. NT-3 supports survival of neurons that differentiate primarily into proprioceptors whereas nerve growth factor and brain-derived neurotrophic factor (BDNF) support subpopulations that transmit nociception and mechanoreception, respectively. We examined sensory neurons of gene-targeted mouse mutants at the NT-4, BDNF, NT-3, and TrkA loci. We show that NT-4 functions early in gangliogenesis, upstream of BDNF. In the absence of NT-4 function, BDNF-dependent, TrkB-expressing neurons fail to appear. The results are consistent with the model that precursor cells intended to become BDNF-dependent mechanoreceptors instead differentiate into NT-3-dependent proprioceptive neurons. C1 Univ Texas, SW Med Ctr, Ctr Dev Biol, Dallas, TX 75235 USA. NCI, Frederick Canc Res & Dev Ctr, ABL Basic Res Program, Neural Dev Grp, Frederick, MD 21702 USA. RP Parada, LF (reprint author), Univ Texas, SW Med Ctr, Ctr Dev Biol, 6000 Harry Hines Blvd, Dallas, TX 75235 USA. RI Parada, luis/B-9400-2014 FU NIGMS NIH HHS [T32 GM008203, T32GM08203]; NINDS NIH HHS [R01 NS 33199, R37 NS033199] NR 44 TC 45 Z9 50 U1 1 U2 1 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD FEB 29 PY 2000 VL 97 IS 5 BP 2297 EP 2302 DI 10.1073/pnas.040562597 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 289QJ UT WOS:000085633200062 PM 10681461 ER PT J AU Bonvini, P Hwang, SG El-Gamil, M Robbins, P Kim, JS Trepel, J Neckers, L AF Bonvini, P Hwang, SG El-Gamil, M Robbins, P Kim, JS Trepel, J Neckers, L TI Nuclear beta-catenin displays GSK-3 beta- and APC-independent proteasome sensitivity in melanoma cells SO BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH LA English DT Article DE beta-catenin; melanoma; proteasome; nuclear cytoskeleton ID ADHESION MOLECULE UVOMORULIN; TRANSCRIPTION FACTOR LEF-1; TUMOR-SUPPRESSOR PROTEIN; ALPHA-CATENIN; POOR-PROGNOSIS; E-CADHERIN; COMPLEX; EXPRESSION; CANCER; LOCALIZATION AB Colon carcinoma and melanoma cells containing either a deletion of the adenomatous polyposis coli tumor suppressor protein (APC) or mutation of the site in beta-catenin phosphorylated by glycogen synthase kinase-3 beta (GSK-3 beta) display elevated levels of detergent-soluble beta-catenin due to insensitivity of the cytosolic protein to proteasome-dependent degradation. In this study, we have examined the effect of beta-catenin mutation (S37F) or APC loss on the proteasome sensitivity of additional subcellular beta-catenin pools in melanoma cells. In contrast to detergent-soluble beta-catenin, the detergent-insoluble protein remains proteasome-sensitive irrespective of S37F mutation or APC status. This insoluble component appears associated primarily with nuclear cytoskeletal elements. In addition, DNase I treatment solubilized a portion of detergent-insoluble beta-catenin, suggesting that this fraction also contains chromatin-associated protein, and correlating with a proteasome-sensitive elevation in beta-catenin-stimulated reporter activity. Since the detergent-insoluble nuclear component of beta-catenin displays GSK-3 beta- and APC-indepcndent proteasome sensitivity, distinct from the soluble nuclear and cytosolic pools of this protein, regulation of beta-catenin proteasome sensitivity and the contribution of this process to beta-catenin function may be more complex than previously appreciated. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NCI, Dept Cell & Canc Biol, Med Branch, NIH, Rockville, MD 20850 USA. NCI, Dept Cell & Canc Biol, Med Branch, NIH, Bethesda, MD 20892 USA. NCI, Surg Branch, NIH, Bethesda, MD 20892 USA. RP Neckers, L (reprint author), NCI, Dept Cell & Canc Biol, Med Branch, NIH, 9610 Med Ctr Dr,Suite 300, Rockville, MD 20850 USA. EM len@helix.nih.gov NR 38 TC 10 Z9 10 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0167-4889 J9 BBA-MOL CELL RES JI Biochim. Biophys. Acta-Mol. Cell Res. PD FEB 28 PY 2000 VL 1495 IS 3 BP 308 EP 318 DI 10.1016/S0167-4889(99)00162-7 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 292UR UT WOS:000085813900011 PM 10699468 ER PT J AU Huang, W Galdzicki, Z van Gelderen, P Balbo, A Chikhale, EG Schapiro, MB Rapoport, SI AF Huang, W Galdzicki, Z van Gelderen, P Balbo, A Chikhale, EG Schapiro, MB Rapoport, SI TI Brain myo-inositol level is elevated in Ts65Dn mouse and reduced after lithium treatment SO NEUROREPORT LA English DT Article DE Down syndrome; lithium; magnetic resonance spectroscopy; myo-inositol; N-acetylaspartate; Ts65Dn ID DOWN-SYNDROME; BEHAVIORAL-ASSESSMENT; IN-VIVO; MODEL; MYOINOSITOL; SPECTROSCOPY; TRISOMY-16; NEURONS; PROTEIN AB The segmental trisomy Ts65Dn mouse is a novel model of Down syndrome (DS). The purpose of this study was to measure brain levels of myo-inositol (mI), N-acetylaspartate (NAA), and other metabolites in Ts65Dn mice using in vivo H-1 magnetic resonance spectroscopy (MRS), and to determine whether lithium (Li) treatment alters brain mI level. The ratio of mi over total creatine (Cr), ml/Cr, was significantly elevated (mean change +38%), while NAA/Cr was significantly decreased (mean change -18%) in Ts65Dn mice (n = 5) compared with control mice (n = 7). This is consistent with H-1 MRS findings in DS human adults. Brain ml/Cr of the entire sample group (n = 12) was reduced (mean change -15%) following Li treatment, supporting the Li-induced ml depletion hypothesis. NeuroReport 11:445-448 (C) 2000 Lippincott Williams & Wilkins. C1 NIA, Neurosci Lab, Bethesda, MD 20892 USA. NINDS, In Vivo NMR Res Ctr, Bethesda, MD 20892 USA. SUNY Stony Brook, Dept Radiol, Stony Brook, NY 11794 USA. RP Huang, W (reprint author), NIA, Neurosci Lab, Bethesda, MD 20892 USA. NR 25 TC 23 Z9 23 U1 1 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0959-4965 J9 NEUROREPORT JI Neuroreport PD FEB 28 PY 2000 VL 11 IS 3 BP 445 EP 448 DI 10.1097/00001756-200002280-00004 PG 4 WC Neurosciences SC Neurosciences & Neurology GA 287CJ UT WOS:000085486900006 PM 10718292 ER PT J AU Kish, LB Bezrukov, SM AF Kish, LB Bezrukov, SM TI Flows of cars and neural spikes enhanced by colored noise SO PHYSICS LETTERS A LA English DT Article ID STOCHASTIC RESONANCE; SYSTEMS AB The problem of interaction between cars at road junctions has been studied for a long time. It is well-known that the interaction generally decreases the traffic efficiency and that randomness in the car flow can help cross-junction traffic efficiency. We show that proper long-range correlations in the flow noise, related to 1/f-like noises, provide superior traffic properties as compared to Poissonian or periodic car traffic. Moreover, a stochastic resonance phenomenon sensitive to the shape of the spectrum occurs. By a small modification, the model can be made relevant for neuronal spike traffic. Spike trains generated by Gaussian 1/f noise show superior spike traffic properties and efficiency of information transfer. (C) 2000 Published by Elsevier Science B.V. All rights reserved. C1 NICHD, Lab Phys & Struct Biol, NIH, Bethesda, MD 20892 USA. Uppsala Univ, Angstrom Lab, SE-75121 Uppsala, Sweden. St Petersburg Nucl Phys Inst, Gatchina 188350, Russia. RP Bezrukov, SM (reprint author), NICHD, Lab Phys & Struct Biol, NIH, Bethesda, MD 20892 USA. NR 21 TC 8 Z9 8 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0375-9601 J9 PHYS LETT A JI Phys. Lett. A PD FEB 28 PY 2000 VL 266 IS 4-6 BP 271 EP 275 DI 10.1016/S0375-9601(00)00058-X PG 5 WC Physics, Multidisciplinary SC Physics GA 291YQ UT WOS:000085767700010 ER PT J AU Hutchinson, ML Schiffman, M AF Hutchinson, ML Schiffman, M TI Utility of liquid-based cytology for cervical carcinoma screening - Reply SO CANCER CYTOPATHOLOGY LA English DT Letter C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. NR 6 TC 0 Z9 0 U1 1 U2 1 PU JOHN WILEY & SONS INC PI NEW YORK PA 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0008-543X J9 CANCER CYTOPATHOL JI Cancer Cytopathol. PD FEB 25 PY 2000 VL 90 IS 1 BP 68 EP 69 PG 2 WC Oncology; Pathology SC Oncology; Pathology GA 286GM UT WOS:000085435300011 ER PT J AU Watari, H Blanchette-Mackie, EJ Dwyer, NK Sun, GS Glick, JM Patel, S Neufeld, EB Pentchev, PG Strauss, JF AF Watari, H Blanchette-Mackie, EJ Dwyer, NK Sun, GS Glick, JM Patel, S Neufeld, EB Pentchev, PG Strauss, JF TI NPC1-containing compartment of human granulosa-lutein cells: A role in the intracellular trafficking of cholesterol supporting steroidogenesis SO EXPERIMENTAL CELL RESEARCH LA English DT Article ID LOW-DENSITY LIPOPROTEIN; HUMAN CHORIONIC-GONADOTROPIN; PLASMA-MEMBRANE CHOLESTEROL; ACUTE REGULATORY PROTEIN; HAMSTER OVARY CELLS; LEYDIG TUMOR-CELLS; 8-BROMO-ADENOSINE 3',5'-MONOPHOSPHATE; TRANSPORT; 3-BETA-<2-(DIETHYLAMINO)ETHOXY>ANDROST-5-EN-17-ONE; METABOLISM AB Steroidogenic cells represent unique systems for the exploration of intracellular cholesterol trafficking. We employed cytochemical and biochemical methods to explore the expression, regulation, and function of the Niemann-Pick C1 protein (NPC1) in human granulosalutein cells. NPC1 was localized in a subset of lysosome-associated membrane glycoprotein 2 (LAMP-2)-positive vesicles. By analyzing the sensitivity of NPC1 N-linked oligosaccharide chains to glycosidases and neuraminidase, evidence was obtained for movement of nascent NPC1 from the endoplasmic reticulum through the medial and trans compartments of the Golgi apparatus prior to its appearance in cytoplasmic vesicles. NPC1 protein content and the morphology and cellular distribution of NPC1-containing vesicles were not affected by treatment of the granulosa-lutein cells with 8-Br-cAMP, which stimulates cholesterol metabolism into progesterone. In contrast, steroidogenic acute regulatory (StAR) protein levels were increased by 8-Br-cAMP. Incubation of granulosa-lutein cells with low-density lipoprotein (LDL) in the presence of the hydrophobic amine, U18666A, caused accumulation of free cholesterol in granules, identified by filipin staining, that contained LAMP-2 and NPC1. These granules also stained for neutral lipid with Nile red, reflecting accumulation of LDL-derived cholesterol esters. LDL-stimulated progesterone synthesis was completely blocked by U18666A, leaving steroid output at levels similar to those of cells incubated in the absence of LDL. The hydrophobic amine also blocked the LDL augmentation of 8-Br-cAMP-stimulated progesterone synthesis, reducing steroid production to levels seen in cells stimulated with 8-Br-cAMP in the absence of LDL. Steroidogenesis recovered after U18666A was removed from the culture medium. UI8666A treatment caused a 2-fold or more increase in NPC1 protein and mRNA levels, suggesting that disruption of NPC1's function activates a compensatory mechanism resulting in increased NPC1 synthesis. We conclude that the NPC1 compartment plays an important role in the trafficking of LDL-derived substrate in steroidogenic cells; that NPC1 expression is up regulated when NPC1 action is blocked; and that the NPC1 compartment can be functionally separated from other intracellular pathways contributing substrate for steroidogenesis. (C) 2000 Academic Press. C1 Univ Penn, Med Ctr, Dept Obstet & Gynecol & Cellular, Ctr Res Reprod & Womens Hlth, Philadelphia, PA 19104 USA. NIDDKD, Sect Lipid Cell Biol, NIH, Bethesda, MD 20892 USA. NINCDS, Cellular & Mol Pathophysiol Sect, Dev & Metab Neurol Branch, NIH, Bethesda, MD 20892 USA. Vet Adm Connecticut Healthcare Syst, Neurol Res Lab, Newington, CT 06111 USA. RP Strauss, JF (reprint author), 1354 BRB 2-3,421 Curie Blvd, Philadelphia, PA 19038 USA. FU NICHD NIH HHS [HD06274]; NINDS NIH HHS [NS34339] NR 42 TC 36 Z9 36 U1 0 U2 5 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0014-4827 J9 EXP CELL RES JI Exp. Cell Res. PD FEB 25 PY 2000 VL 255 IS 1 BP 56 EP 66 DI 10.1006/excr.1999.4774 PG 11 WC Oncology; Cell Biology SC Oncology; Cell Biology GA 290FR UT WOS:000085666100008 PM 10666334 ER PT J AU Bagby, S Mal, TK Liu, DJ Raddatz, E Nakatani, Y Ikura, M AF Bagby, S Mal, TK Liu, DJ Raddatz, E Nakatani, Y Ikura, M TI TFIIA-TAF regulatory interplay: NMR evidence for overlapping binding sites on TBP SO FEBS LETTERS LA English DT Article DE TATA box binding protein; TBP-associated factor; transcription factor IIA; nuclear magnetic resonance; heteronuclear single quantum coherence ID RNA-POLYMERASE-II; TATA-BOX-BINDING; TRANSCRIPTION FACTOR TFIIA; YEAST TFIIA/TBP/DNA COMPLEX; CRYSTAL-STRUCTURE; IN-VIVO; ACTIVATION DOMAIN; MINOR-GROOVE; DNA-BINDING; PROTEIN AB TATA box binding protein (TBP)-promoter interaction nucleates assembly of the RNA polymerase II transcription initiation complex. Transcription factor IIA (TFIIA) stabilizes the TBP-promoter complex whereas the N-terminal domain of the largest TAF(II) inhibits TBP-promoter interaction. We have mapped the interaction sites on TBP of Drosophila TAF(II)230 and yeast TFIIA (comprising two subunits, TOA1 and TOA2), using nuclear magnetic resonance (NMR), and also report structural evidence that subdomain II of the TAF(II)230 N-terminal inhibitory domain and TFIIA have overlapping binding sites on the convex surface of TBP, Together with previous mutational and biochemical data, our NMR results indicate that subdomain II augments subdomain I-mediated inhibition of TBP function by blocking TBP-TFIIA interaction. (C) 2000 Federation of European Biochemical Societies. C1 Univ Toronto, Div Mol & Struct Biol, Ontario Canc Inst, Toronto, ON M5G 2M9, Canada. Univ Toronto, Dept Med Biophys, Toronto, ON M5G 2M9, Canada. NICHHD, Lab Mol Growth Regulat, NIH, Bethesda, MD 20892 USA. RP Ikura, M (reprint author), Univ Toronto, Div Mol & Struct Biol, Ontario Canc Inst, 610 Univ Ave, Toronto, ON M5G 2M9, Canada. NR 40 TC 25 Z9 26 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 FEB 25 PY 2000 VL 468 IS 2-3 BP 149 EP 154 DI 10.1016/S0014-5793(00)01213-8 PG 6 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 290DY UT WOS:000085662000009 PM 10692576 ER PT J AU Mitchell, DC Litman, BJ AF Mitchell, DC Litman, BJ TI Effect of ethanol and osmotic stress on receptor conformation - Reduced water activity amplifies the effect of ethanol on metarhodopsin II formation SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID POLY(ETHYLENE GLYCOL); BOVINE RHODOPSIN; CHOLESTEROL; MECHANISM; MEMBRANES; ORDER; PHOSPHOLIPIDS; UNSATURATION; EQUILIBRIUM; ANESTHESIA AB The combined effects of ethanol and osmolytes on both the extent of formation of metarhodopsin II (MII), which binds and activates transducin, and on acyl chain packing were examined in rod outer segment disc membranes. The ethanol-induced increase in MII formation was amplified by the addition of neutral osmolytes. This enhancement was linear with osmolality, At 360 millios-molal, the osmolality of human plasma, 50 mM ethanol was 2.7 times more potent than at 0 osmolality, demonstrating the importance of water activity in in vitro experiments dealing with ethanol potency. Ethanol disordered acyl chain packing, and increasing osmolality enhanced this acyl chain disordering. Prior osmotic stress data showed a release of 35 +/- 2 water molecules upon MII formation. Ethanol increases this number to 49 water molecules, suggesting that ethanol replaces 15 additional water molecules upon MII formation. Amplification of ethanol effects on MII formation and acyl chain packing by osmolytes suggests that ethanol increases the equilibrium concentration of MII both by disordering acyl chain packing and by disrupting rhodopsin-water hydrogen bonds, demonstrating a direct effect of ethanol on rhodopsin. At physiologically relevant levels of osmolality and ethanol, about 90% of ethanol's effect is due to disordered acyl chain packing. C1 NIAAA, Sect Fluorescence Studies, Lab Membrane Biophys & Biochem, NIH, Rockville, MD 20853 USA. RP Litman, BJ (reprint author), Pk Bldg,Room 158,12420 Parklawn Dr, Rockville, MD 20852 USA. NR 33 TC 33 Z9 33 U1 1 U2 5 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD FEB 25 PY 2000 VL 275 IS 8 BP 5355 EP 5360 DI 10.1074/jbc.275.8.5355 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 286XQ UT WOS:000085474500016 PM 10681509 ER PT J AU Takeda, A Perry, G Abraham, NG Dwyer, BE Kutty, RK Laitinen, JT Petersen, RB Smith, MA AF Takeda, A Perry, G Abraham, NG Dwyer, BE Kutty, RK Laitinen, JT Petersen, RB Smith, MA TI Overexpression of heme oxygenase in neuronal cells, the possible interaction with tau SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID TRANSIENT FOREBRAIN ISCHEMIA; RAT-BRAIN; ALZHEIMERS-DISEASE; MESSENGER-RNA; OXIDATIVE STRESS; CARBON-MONOXIDE; PC12 CELLS; PROTEIN; INDUCTION; EXPRESSION AB Increased expression of heme oxygenase-1 (HO-1) is a common feature in a number of neurodegenerative diseases. Interestingly, the spatial distribution of HO-1 expression in diseased brain is essentially identical to that of pathological expression of tau. In this study, we explored the relationship between HO-1 and tau, using neuroblastoma cells stably transfected with sense and antisense HO-1 constructs as well as with the vector alone. In transfected cells overexpressing HO-1, the activity of heme oxygenase was increased, and conversely, the level of tau protein was dramatically decreased when compared with antisense HO-1 or CEP transfected cells. The suppression of tau protein expression was almost completely reversed by zinc-deuteroporphyrin, a specific inhibitor of heme oxygenase activity. The activated forms of ERKs (extracellular signal-regulated kinases) were also decreased in cells overexpressing HO-I although no changes in the expression of total ERB-1/2 proteins were observed. These data are in agreement with the finding that the expression of tau is regulated through signal cascades including the ERKs, whose activities are modulated by oxidative stresses. The expression of tan and HO-1 may be regulated by oxidative stresses in a coordinated manner and play a pivotal role in the cytoprotection of neuronal cells. C1 Case Western Reserve Univ, Inst Pathol, Cleveland, OH 44106 USA. New York Med Coll, Dept Pharmacol, Valhalla, NY 10595 USA. Vet Affairs Med Ctr, Res Serv, White River Junction, VT 05009 USA. NEI, NIH, Bethesda, MD 20892 USA. Univ Kuopio, Dept Physiol, FIN-70211 Kuopio, Finland. RP Smith, MA (reprint author), Case Western Reserve Univ, Inst Pathol, 2085 Adelbert Rd, Cleveland, OH 44106 USA. RI Smith, Mark/A-9053-2009; Perry, George/A-8611-2009; Petersen, Robert/B-5075-2011 OI Perry, George/0000-0002-6547-0172; Petersen, Robert/0000-0002-3154-0072 FU NIA NIH HHS [AG09287, AG14249]; NINDS NIH HHS [NS38648] NR 41 TC 134 Z9 144 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD FEB 25 PY 2000 VL 275 IS 8 BP 5395 EP 5399 DI 10.1074/jbc.275.8.5395 PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 286XQ UT WOS:000085474500021 PM 10681514 ER PT J AU Graves, PR Yu, LJ Schwarz, JK Gales, J Sausville, EA O'Connor, PM Piwnica-Worms, H AF Graves, PR Yu, LJ Schwarz, JK Gales, J Sausville, EA O'Connor, PM Piwnica-Worms, H TI The Chk1 protein kinase and the Cdc25C regulatory pathways are targets of the anticancer agent UCN-01 SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID CELL-CYCLE REGULATION; TYROSINE KINASE; DNA-DAMAGE; NUCLEAR EXPORT; FISSION YEAST; G(2) ARREST; HUMAN WEE1; PHOSPHORYLATES CDC2; CHECKPOINT PATHWAY; DEPENDENT KINASES AB A checkpoint operating in the G(2) phase of the cell cycle prevents entry into mitosis in the presence of DNA damage, UCN-01, a protein kinase inhibitor currently undergoing clinical trials for cancer treatment, abrogates G(2) checkpoint function and sensitizes p53-defective cancer cells to DNA damaging agents. In most species, the G(2) checkpoint prevents the Cdc25 phosphatase from removing inhibitory phosphate groups from the mitosis-promoting kinase Cdc2. This is accomplished by maintaining Cdc25 in a phosphorylated form that binds 14-3-3 proteins. The checkpoint kinases, Chk1 and Cds1, are proposed to regulate the interactions between human Cdc25C and 14-3-3 proteins by phosphorylating Cdc25C on serine 216. 14-3-3 proteins, in turn, function to keep Cdc25C out of the nucleus. Here we report that UCN-01 caused loss of both serine 216 phosphorylation and 14-3-3 binding to Cdc25C in DNA-damaged cells. In addition, UCN-01 potently inhibited the ability of Chk1 to phosphorylate Cdc25C in vitro. In contrast, Cds1 was refractory to inhibition by UCN-01 in vitro, and Cds1 was still phosphorylated in irradiated cells treated with UCN-01. Thus, neither Cds1 nor kinases upstream of Cds1, such as ataxia telangiectasia-mutated, are targets of UCN-01 action in vivo. Taken together our results identify the Chk1 kinase and the Cdc25C pathway as potential targets of G(2) checkpoint abrogation by UCN-01. C1 Washington Univ, Sch Med, Howard Hughes Med Inst, Dept Cell Biol & Physiol, St Louis, MO 63110 USA. NCI, Mol Pharmacol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. NCI, Dev Therapeut Program, Div Canc Treatment & Diagnosis, NIH, Bethesda, MD 20892 USA. RP Piwnica-Worms, H (reprint author), Washington Univ, Sch Med, Howard Hughes Med Inst, Dept Cell Biol & Physiol, Box 8228,660 S Euclid Ave, St Louis, MO 63110 USA. RI Piwnica-Worms, Helen/C-5214-2012 NR 65 TC 467 Z9 475 U1 1 U2 13 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD FEB 25 PY 2000 VL 275 IS 8 BP 5600 EP 5605 DI 10.1074/jbc.275.8.5600 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 286XQ UT WOS:000085474500048 PM 10681541 ER PT J AU Sanchez, LB Galperin, MY Muller, M AF Sanchez, LB Galperin, MY Muller, M TI Acetyl-CoA synthetase from the amitochondriate eukaryote Giardia lamblia belongs to the newly recognized superfamily of acyl-CoA synthetases (nucleoside diphosphate-forming) SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID ESCHERICHIA-COLI; PYROCOCCUS-FURIOSUS; ACETATE FORMATION; TRICHOMONAS-VAGINALIS; SUCCINATE THIOKINASE; PYRUVATE METABOLISM; ATP SYNTHESIS; COENZYME-A; PURIFICATION; EXPRESSION AB The gene coding for the acetyl-CoA. synthetase (ADP-forming) from the amitochondriate eukaryote Giardia lamblia has been expressed in Escherichia coli. The recombinant enzyme exhibited the same substrate specificity as the native enzyme, utilizing acetyl-CoA and adenine nucleotides as preferred substrates and less efficiently, propionyl- and succinyl-CoA. N- and C-terminal parts of the G. lamblia acetyl-CoA sythetase sequence were found to be homologous to the alpha- and beta-subunits, respectively, of succinyl-CoA synthetase, Sequence analysis of homologous enzymes from various bacteria archaea, and the eukaryote, Plasmodium fad ciparum, identified conserved features in their organization, which allowed us to delineate a new superfamily of acyl-CoA synthetases (nucleoside diphosphate-forming) and its signature motifs, The representatives of this new superfamily of thiokinases vary in their domain arrangement, some consisting of separate alpha- and beta-subunits and others comprising fusion proteins in alpha-beta or beta-alpha orientation. The presence of homologs of acetyl synthetase (ADP-forming) in such human pathogens as G. lamblia, Yersinia pestis, Bordetella pertussis, Pseudomonas aeruginosa, Vibrio cholerae, Salmonella typhi, Porphyromonas gingivalis, and the malaria agent P. falciparum suggests that they might be used as potential drug targets. C1 Rockefeller Univ, New York, NY 10021 USA. Natl Lib Med, Natl Ctr Biotechnol Informat, NIH, Bethesda, MD 20894 USA. RP Sanchez, LB (reprint author), Rockefeller Univ, 1230 York Ave, New York, NY 10021 USA. RI Galperin, Michael/B-5859-2013 OI Galperin, Michael/0000-0002-2265-5572 FU NIAID NIH HHS [AI11942] NR 44 TC 68 Z9 70 U1 1 U2 5 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD FEB 25 PY 2000 VL 275 IS 8 BP 5794 EP 5803 DI 10.1074/jbc.275.8.5794 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 286XQ UT WOS:000085474500075 PM 10681568 ER PT J AU Joseph, GT Huima, T Klion, A Lustigman, S AF Joseph, GT Huima, T Klion, A Lustigman, S TI A novel developmentally regulated galectin of Onchocerca volvulus SO MOLECULAR AND BIOCHEMICAL PARASITOLOGY LA English DT Article DE Onchocerca volvulus; larvae; galectin; putatively immune individuals; antigen ID GALACTOSIDE-BINDING LECTINS; PROTECTIVE IMMUNITY; MESSENGER-RNAS; ANIMAL LECTINS; CDNA CLONING; SEQUENCE; ELEGANS; PROTEIN; FAMILY; STAGE C1 New York Blood Ctr, Lindsley F Kimball Res Inst, Mol Parasitol Lab, New York, NY 10021 USA. NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. RP Lustigman, S (reprint author), New York Blood Ctr, Lindsley F Kimball Res Inst, Mol Parasitol Lab, 310 E 67th St, New York, NY 10021 USA. OI Klion, Amy/0000-0002-4986-5326 NR 26 TC 10 Z9 10 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0166-6851 J9 MOL BIOCHEM PARASIT JI Mol. Biochem. Parasitol. PD FEB 25 PY 2000 VL 106 IS 1 BP 187 EP 195 DI 10.1016/S0166-6851(99)00208-X PG 9 WC Biochemistry & Molecular Biology; Parasitology SC Biochemistry & Molecular Biology; Parasitology GA 288GC UT WOS:000085553500019 PM 10743624 ER PT J AU Song, S Sorimachi, K Adachi, K Oka, T AF Song, S Sorimachi, K Adachi, K Oka, T TI Biochemical and molecular biological evidence for the presence of type II iodothyronine deiodinase in mouse mammary gland SO MOLECULAR AND CELLULAR ENDOCRINOLOGY LA English DT Article DE type II iodothyronine deiodinase; mouse mammary gland; thyroid hormone; reproductive stage ID THYROXINE 5'-DEIODINASE ACTIVITY; THYROID-HORMONES; LACTATING RATS; EXPRESSION; SELENOCYSTEINE; SELENOPROTEIN; DEFICIENCY; INVITRO; CLONING; PROPYLTHIOURACIL AB In the present study we have obtained several lines of evidence indicating the presence of type II iodothyronine deiodinase (DII) in the mouse mammary gland. 5'-deiodinase activity in the mammary gland has an apparent K-m value of 4.4 nM for T-4 and is inhibited by aurothioglucose but not by propylthiouracil. These characteristics are similar to those of DII in other tissues. We cloned a 1.4-kb cDNA, which contains the entire mouse DII coding region and has high homology with the rat DII cDNA, from the mammary gland and brain. Northern blot analysis showed the presence of 7.9 kb DII mRNA in the mammary gland and brain. The levels of DII activity and mRNA in lactating gland were significantly lower than those in virgin and pregnant glands, suggesting that DII is regulated at the pretranslational level. In addition, we found the low level of DII enzyme activity and transcript in various other mouse tissues. Published by Elsevier Science Ireland Ltd. C1 NIDDK, Lab Genet & Physiol, NIH, Bethesda, MD 20892 USA. RP Oka, T (reprint author), NIDDK, Lab Genet & Physiol, NIH, Bldg 8,Rm 106,9000 Rockville Pike, Bethesda, MD 20892 USA. OI Sorimachi, Kenji/0000-0002-5259-9671 NR 50 TC 14 Z9 15 U1 0 U2 2 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0303-7207 J9 MOL CELL ENDOCRINOL JI Mol. Cell. Endocrinol. PD FEB 25 PY 2000 VL 160 IS 1-2 BP 173 EP 181 DI 10.1016/S0303-7207(99)00249-X PG 9 WC Cell Biology; Endocrinology & Metabolism SC Cell Biology; Endocrinology & Metabolism GA 298GQ UT WOS:000086131300019 PM 10715551 ER PT J AU Meng, XJ Lindahl, M Hyvonen, ME Parvinen, M de Rooij, DG Hess, MW Raatikainen-Ahokas, A Sainio, K Rauvala, H Lakso, M Pichel, JG Westphal, H Saarma, M Sariola, H AF Meng, XJ Lindahl, M Hyvonen, ME Parvinen, M de Rooij, DG Hess, MW Raatikainen-Ahokas, A Sainio, K Rauvala, H Lakso, M Pichel, JG Westphal, H Saarma, M Sariola, H TI Regulation of cell fate decision of undifferentiated spermatogonia by GDNF SO SCIENCE LA English DT Article ID ENTERIC NERVOUS-SYSTEM; ACID RECEPTOR-ALPHA; MICE LACKING GDNF; NEUROTROPHIC FACTOR; IN-VIVO; MOTOR-NEURONS; EXPRESSION; TESTIS; SPERMATOGENESIS; MOUSE AB The molecular control of self-renewal and differentiation of stem cells has remained enigmatic. Transgenic Loss-of-function and overexpression models now show that the dosage of glial cell Line-derived neurotrophic factor (GDNF), produced by Sertoli cells, regulates cell fate decisions of undifferentiated spermatogonial cells that include the stem cells for spermatogenesis. Gene-targeted mice with one GDNF-null allele show depletion of stem cell reserves, whereas mice overexpressing GDNF show accumulation of undifferentiated spermatogonia. They are unable to respond properly to differentiation signals and undergo apoptosis upon retinoic acid treatment. Nonmetastatic testicular tumors are regularly formed in older GDNF-overexpressing mice. Thus, GDNF contributes to paracrine regulation of spermatogonial self-renewal and differentiation. C1 Univ Helsinki, Inst Biotechnol, Viikki Bioctr, Res Program Dev Biol, FIN-00014 Helsinki, Finland. Univ Helsinki, Inst Biotechnol, Viikki Bioctr, Res Program Mol Neurobiol, FIN-00014 Helsinki, Finland. Univ Helsinki, Inst Biotechnol, Transgen Unit, FIN-00014 Helsinki, Finland. Univ Helsinki, Dept Biosci, FIN-00014 Helsinki, Finland. Univ Turku, Dept Anat, FIN-20520 Turku, Finland. Univ Utrecht, Med Ctr, Dept Cell Biol, NL-3584 CX Utrecht, Netherlands. Hosp Merida, Unidad Invest, Badajoz 06800, Spain. NICHHD, Bethesda, MD 20892 USA. RP Sariola, H (reprint author), Univ Helsinki, Inst Biotechnol, Viikki Bioctr, Res Program Dev Biol, POB 56,Viikinkaari 9, FIN-00014 Helsinki, Finland. OI de Rooij, Dirk/0000-0003-3932-4419 NR 39 TC 680 Z9 767 U1 6 U2 39 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 USA SN 0036-8075 J9 SCIENCE JI Science PD FEB 25 PY 2000 VL 287 IS 5457 BP 1489 EP 1493 DI 10.1126/science.287.5457.1489 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 287WZ UT WOS:000085531600052 PM 10688798 ER PT J AU Priola, SA Raines, A Caughey, WS AF Priola, SA Raines, A Caughey, WS TI Porphyrin and phthalocyanine antiscrapie compounds SO SCIENCE LA English DT Article ID RESISTANT PRION PROTEIN; HAMSTER SCRAPIE; AMPHOTERICIN-B; VARIANT CJD; CONGO RED; PRP; INHIBITION; AGENT; PATHOGENESIS; EXPRESSION AB The transmissible spongiform encephalopathies (TSEs) are fatal, neurodegenerative diseases for which no effective treatments are available. The Likelihood that a bovine form of TSE has crossed species barriers and infected humans underscores the urgent need to identify anti-TSE drugs. Certain cyclic tetrapyrroles (porphyrins and phthalocyanines) have recently been shown to inhibit the in vitro formation of PrP-res, a protease-resistant protein critical for TSE pathogenesis. We now report that treatment of TSE-infected animals with three such compounds increased survival time from 50 to 300%. The significant inhibition of TSE disease by structurally dissimilar tetrapyrroles identifies these compounds as anti-TSE drugs. C1 NIAID, Rocky Mt Labs, Persistent Viral Dis Lab, NIH, Hamilton, MT 59840 USA. RP Priola, SA (reprint author), NIAID, Rocky Mt Labs, Persistent Viral Dis Lab, NIH, Hamilton, MT 59840 USA. FU NIA NIH HHS [AG04342] NR 26 TC 235 Z9 243 U1 1 U2 25 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 USA SN 0036-8075 J9 SCIENCE JI Science PD FEB 25 PY 2000 VL 287 IS 5457 BP 1503 EP 1506 DI 10.1126/science.287.5457.1503 PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 287WZ UT WOS:000085531600056 PM 10688802 ER PT J AU Kasuya, J Liang, SJ Goko, H Park, SH Kato, K Xu, ZD Hockman, S Manganiello, VC Fujita-Yamaguchi, Y AF Kasuya, J Liang, SJ Goko, H Park, SH Kato, K Xu, ZD Hockman, S Manganiello, VC Fujita-Yamaguchi, Y TI Cardiac type cGMP-inhibited phosphodiesterase (PDE3A) gene structure: Similarity and difference to adipocyte type PDE3B gene SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article ID CYCLIC-NUCLEOTIDE PHOSPHODIESTERASE; CAMP-PHOSPHODIESTERASE; MOLECULAR-CLONING; PURIFICATION; EXPRESSION; ISOFORM; CDNA AB Phosphodiesterase type 3 isoforms, PDE3A and 3B, are expressed primarily in cardiovascular and adipose tissues, respectively. We previously reported a shorter transcript of 4.4-kb PDE3A which is predominantly transcribed in human placenta, whereas a full-length 7.6-kb transcript corresponding to the cardiac PDE3A cDNA has not been characterized. Due to unfortunate circumstances created by changes in PDE3 nomenclature, PDE3B gene structure previously reported used PDE3A in its title. Here, we describe PDE3A gene structure, which comprises 16 exons spanning over 130 kb on chromosome 12p12. Two PDE3 isoforms share similar gene organization, but localize to different chromosomes. The most distal transcription initiation site of the PDE3A gene is similar to 1071 bases upstream of the ATG: site, suggesting that exon 1 consists of 1071 and 960 bp of untranslated and translated sequences, respectively. The proximal 5'-flanking region, which does not contain TATA-like sequences, exhibited weak but significant promoter activity. Results suggest potential involvement of distal promoter/enhancer and translational regulation for expression of the 7.6-kb transcript. (C) 2000 Academic Press. C1 Beckman Res Inst, Dept Biol Mol, Duarte, CA 91010 USA. NHLBI, Pulm Crit Care Med Branch, NIH, Bethesda, MD 20892 USA. RP Fujita-Yamaguchi, Y (reprint author), Beckman Res Inst, Dept Biol Mol, Duarte, CA 91010 USA. FU NCI NIH HHS [CA65767] NR 23 TC 13 Z9 14 U1 0 U2 1 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD FEB 24 PY 2000 VL 268 IS 3 BP 827 EP 834 DI 10.1006/bbrc.2000.2226 PG 8 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 289UH UT WOS:000085640000031 PM 10679291 ER PT J AU Aravind, L Koonin, EV AF Aravind, L Koonin, EV TI The U box is a modified RING finger - a common domain in ubiquitination SO CURRENT BIOLOGY LA English DT Letter ID PROTEIN FOLD RECOGNITION; ZINC-FINGER; LIGASE; SYSTEM C1 NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA. RP Aravind, L (reprint author), NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA. NR 19 TC 154 Z9 166 U1 2 U2 24 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0960-9822 J9 CURR BIOL JI Curr. Biol. PD FEB 24 PY 2000 VL 10 IS 4 BP R132 EP R134 DI 10.1016/S0960-9822(00)00398-5 PG 3 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 348GR UT WOS:000088977600005 PM 10704423 ER PT J AU Kim, YC Gallo-Rodriguez, C Jang, SY Nandanan, E Adams, M Harden, TK Boyer, JL Jacobson, KA AF Kim, YC Gallo-Rodriguez, C Jang, SY Nandanan, E Adams, M Harden, TK Boyer, JL Jacobson, KA TI Acyclic analogues of deoxyadenosine 3 ',5 '-bisphosphates as P2Y(1) receptor antagonists SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID PHOSPHOLIPASE-C; P2 RECEPTORS; PURINERGIC RECEPTOR; POTENT; IDENTIFICATION; DERIVATIVES; ACTIVATION; EXPRESSION; PLATELETS; AGONISTS AB P2Y(1) receptors are activated by ADP and occur on endothelial cells, smooth muscle, epithelial cells, lungs, pancreas, platelets, and in the central nervous system. With the aid of molecular modeling, we have designed nucleotide analogues that act as selective antagonists at this subtype. The present study has tested the hypothesis that acyclic modifications of the ribose ring, proven highly successful for nucleoside antiviral agents such as gancyclovir, are generalizable to P2Y receptor ligands. Specifically, the binding site of the P2Y(1) receptor was found to be sufficiently accommodating to allow the substitution of the ribose group with acyclic aliphatic and aromatic chains attached to the 9-position of adenine. Three groups of adenine derivatives having diverse side-chain structures, each containing two symmetrical phosphate or phosphonate groups, were prepared. Biological. activity was demonstrated by the ability of the acyclic derivatives to act as agonists or antagonists in the stimulation of phospholipase C in turkey erythrocyte membranes. An acyclic N-6-methyladenine derivative, 2-[2(6-methylaminopurin-9-yl)-ethyl]-propane-1,3-bisoxy(diammoniumphosphate) (10), containing an isopentyl bisphosphate moiety, was a full antagonist at the P2Y(1) receptor with an IC50 value of 1.60 mu M. The corresponding 2-Cl derivative (11) was even more potent with an IC50 value of 0.84 mu M Homologation of the ethylene group at the g-position to 3-5 methylene units or inclusion of cis- or trans-olefinic groups greatly reduced antagonist potency at the P2Y(1) receptor. Analogues containing a diethanolamine amide group and an aryl di(methylphosphonate) were both less potent than 10 as antagonists, with IC50 values of 14 and 16 mu M, respectively, and no agonist activity was observed for these analogues. Thus, the ribose moiety is clearly not essential for recognition by the turkey P2Y(1) receptor, although a cyclic structure appears to be important for receptor activation, and the acyclic approach to the design of P2 receptor antagonists is valid. C1 NIDDK, Mol Recognit Sect, LBC, NIH, Bethesda, MD 20892 USA. Univ N Carolina, Sch Med, Dept Pharmacol, Chapel Hill, NC 27599 USA. Univ Buenos Aires, Fac Ciencias Exactas & Nat, Dept Quim Organ, CIHIDECAR, RA-1428 Buenos Aires, DF, Argentina. RP Jacobson, KA (reprint author), NIDDK, Mol Recognit Sect, LBC, NIH, Bldg 8A,Rm B1A-19, Bethesda, MD 20892 USA. RI Gallo-Rodriguez, Carola/E-1732-2012; Jacobson, Kenneth/A-1530-2009; Erathodiyil, Nandanan/A-8333-2010 OI Jacobson, Kenneth/0000-0001-8104-1493; FU NHLBI NIH HHS [HL54889]; NIGMS NIH HHS [GM38213] NR 30 TC 29 Z9 30 U1 0 U2 2 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0022-2623 J9 J MED CHEM JI J. Med. Chem. PD FEB 24 PY 2000 VL 43 IS 4 BP 746 EP 755 DI 10.1021/jm9905211 PG 10 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 287LR UT WOS:000085507800020 PM 10691699 ER PT J AU Gavrilova, O Marcus-Samuels, B Leon, LR Vinson, C Reitman, ML AF Gavrilova, O Marcus-Samuels, B Leon, LR Vinson, C Reitman, ML TI Hormones - Leptin and diabetes in lipoatrophic mice SO NATURE LA English DT Article ID ADIPOSE-TISSUE; OBESITY C1 NIDDKD, Diabet Branch, NIH, Bethesda, MD 20892 USA. NCI, Lab Metab, NIH, Bethesda, MD 20892 USA. RP Gavrilova, O (reprint author), NIDDKD, Diabet Branch, NIH, Bethesda, MD 20892 USA. RI Reitman, Marc/B-4448-2013 OI Reitman, Marc/0000-0002-0426-9475 NR 9 TC 58 Z9 59 U1 0 U2 4 PU MACMILLAN MAGAZINES LTD PI LONDON PA PORTERS SOUTH, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD FEB 24 PY 2000 VL 403 IS 6772 BP 850 EP 850 PG 1 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 288JG UT WOS:000085559200042 PM 10706272 ER PT J AU Birk, OS Casiano, DE Wassif, CA Cogliati, T Zhao, LP Zhao, YG Grinberg, A Huang, SP Kreidberg, JA Parker, KL Porter, FD Westphal, H AF Birk, OS Casiano, DE Wassif, CA Cogliati, T Zhao, LP Zhao, YG Grinberg, A Huang, SP Kreidberg, JA Parker, KL Porter, FD Westphal, H TI The LIM homeobox gene Lhx9 is essential for mouse gonad formation SO NATURE LA English DT Article ID SEXUAL-DIFFERENTIATION; EXPRESSION AB During mammalian embryonic development, the ovaries and testes develop from somatic cells of the urogenital ridges as indifferent gonads, harbouring primordial germ cells that have migrated there. After sex determination of the gonads, the testes produce testosterone and anti-Mullerian hormone which mediate male sexual differentiation, and the female developmental pathway ensues in their absence(1-3). Here we show that transcripts of the LIM homeobox gene Lhx9 are present in urogenital ridges of mice at embryonic day 9.5; later they localize to the interstitial region as morphological differentiation occurs. In mice lacking Lhx9 function, germ cells migrate normally, but somatic cells of the genital ridge fail to proliferate and a discrete gonad fails to form. In the absence of testosterone and anti-Mullerian hormone, genetically male mice are phenotypically female. The expression of steroidogenic factor 1 (Sf1), a nuclear receptor essential for gonadogenesis2, is reduced to minimal levels in the Lhx9-deficient genital ridge, indicating that Lhx9 may lie upstream of Sf1 in a developmental cascade. Unlike mice lacking other genes that mediate early stages of gonadogenesis(4-6), Lhx9 mutants do not exhibit additional major developmental defects. Thus, LHX9 mutations may underlie certain forms of isolated gonadal agenesis in humans. C1 NICHHD, Lab Mammalian Genes & Dev, NIH, Bethesda, MD 20892 USA. NICHHD, Heritable Disorders Branch, NIH, Bethesda, MD 20892 USA. NCI, Dept Genet, Med Branch, NIH, Bethesda, MD 20889 USA. Univ Texas, SW Med Ctr, Dept Internal Med, Dallas, TX 75235 USA. Childrens Hosp, Dept Med, Boston, MA 02115 USA. RP Westphal, H (reprint author), NICHHD, Lab Mammalian Genes & Dev, NIH, Bethesda, MD 20892 USA. OI Wassif, Christopher/0000-0002-2524-1420 NR 23 TC 187 Z9 194 U1 1 U2 12 PU MACMILLAN MAGAZINES LTD PI LONDON PA PORTERS SOUTH, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD FEB 24 PY 2000 VL 403 IS 6772 BP 909 EP 913 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 288JG UT WOS:000085559200062 PM 10706291 ER PT J AU Carey, JC Klebanoff, MA Hauth, JC Hillier, SL Thom, EA Ernest, JM Heine, RP Nugent, RP Fischer, ML Leveno, KJ Wapner, R Varner, M AF Carey, JC Klebanoff, MA Hauth, JC Hillier, SL Thom, EA Ernest, JM Heine, RP Nugent, RP Fischer, ML Leveno, KJ Wapner, R Varner, M CA Natl Inst Child Hlth Human Dev Net TI Metronidazole to prevent preterm delivery in pregnant women with asymptomatic bacterial vaginosis. SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Article ID LOW-BIRTH-WEIGHT; CONTROLLED TRIAL; INFECTION; PREMATURITY; LABOR AB Background: Bacterial vaginosis has been associated with preterm birth. In clinical trials, the treatment of bacterial vaginosis in pregnant women who previously had a preterm delivery reduced the risk of recurrence. Methods: To determine whether treating women in a general obstetrical population who have asymptomatic bacterial vaginosis (as diagnosed on the basis of vaginal Gram's staining and pH) prevents preterm delivery, we randomly assigned 1953 women who were 16 to less than 24 weeks pregnant to receive two 2-g doses of metronidazole or placebo. The diagnostic studies were repeated and a second treatment was administered to all the women at 24 to less than 30 weeks' gestation. The primary outcome was the rate of delivery before 37 weeks' gestation. Results: Bacterial vaginosis resolved in 657 of 845 women who had follow-up Gram's staining in the metronidazole group (77.8 percent) and 321 of 859 women in the placebo group (37.4 percent). Data on the time and characteristics of delivery were available for 953 women in the metronidazole group and 966 in the placebo group. Preterm delivery occurred in 116 women in the metronidazole group (12.2 percent) and 121 women in the placebo group (12.5 percent) (relative risk, 1.0; 95 percent confidence interval, 0.8 to 1.2). Treatment did not prevent preterm deliveries that resulted from spontaneous labor (5.1 percent in the metronidazole group vs. 5.7 percent in the placebo group) or spontaneous rupture of the membranes (4.2 percent vs. 3.7 percent), nor did it prevent delivery before 32 weeks (2.3 percent vs. 2.7 percent). Treatment with metronidazole did not reduce the occurrence of preterm labor, intraamniotic or postpartum infections, neonatal sepsis, or admission of the infant to the neonatal intensive care unit. Conclusions: The treatment of asymptomatic bacterial vaginosis in pregnant women does not reduce the occurrence of preterm delivery or other adverse perinatal outcomes. (N Engl J Med 2000;342:534-40.) (C)2000, Massachusetts Medical Society. C1 Univ Oklahoma, Dept Obstet & Gynecol, Oklahoma City, OK USA. NICHHD, NIH, Bethesda, MD 20892 USA. Univ Alabama, Dept Obstet & Gynecol, Birmingham, AL 35294 USA. Univ Pittsburgh, Dept Obstet Gynecol & Reprod Sci, Pittsburgh, PA USA. George Washington Univ, Ctr Biostat, Rockville, MD USA. Wake Forest Univ, Bowman Gray Sch Med, Dept Obstet & Gynecol, Winston Salem, NC 27103 USA. Univ Texas, SW Med Ctr, Dept Obstet & Gynecol, Dallas, TX 75235 USA. Thomas Jefferson Univ, Dept Obstet & Gynecol, Philadelphia, PA USA. Univ Utah, Dept Obstet & Gynecol, Salt Lake City, UT USA. Ohio State Univ, Dept Obstet & Gynecol, Columbus, OH 43210 USA. Univ Chicago, Dept Obstet & Gynecol, Chicago, IL 60637 USA. Univ Tennessee, Dept Obstet & Gynecol, Memphis, TN 38103 USA. Univ Cincinnati, Dept Obstet & Gynecol, Cincinnati, OH USA. Wayne State Univ, Dept Obstet & Gynecol, Detroit, MI USA. Univ Miami, Dept Obstet & Gynecol, Miami, FL 33152 USA. Med Univ S Carolina, Dept Obstet & Gynecol, Charleston, SC 29425 USA. RP Klebanoff, MA (reprint author), NICHHD, NIH, 6100 Execut Blvd,Rm 7B03 MSC 7510, Bethesda, MD 20892 USA. RI Varner, Michael/K-9890-2013 OI Varner, Michael/0000-0001-9455-3973 FU NICHD NIH HHS [U10 HD21414, U10 HD21410, U10 HD27869] NR 21 TC 342 Z9 360 U1 0 U2 5 PU MASSACHUSETTS MEDICAL SOC PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD FEB 24 PY 2000 VL 342 IS 8 BP 534 EP 540 DI 10.1056/NEJM200002243420802 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 286AX UT WOS:000085422400002 PM 10684911 ER PT J AU Miotti, PG Taha, TET Kumwenda, NI Van der Hoeven, L Broadhead, R Mtimavalye, LAR Chiphangwi, JD Liomba, G Biggar, RJ AF Miotti, PG Taha, TET Kumwenda, NI Van der Hoeven, L Broadhead, R Mtimavalye, LAR Chiphangwi, JD Liomba, G Biggar, RJ TI Risk of HIV transmission through breastfeeding - Reply SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Letter C1 Johns Hopkins Univ Hosp, Sch Hyg & Publ Hlth, Baltimore, MD 21205 USA. Malawi Coll Med, Blantyre, Malawi. NIH, Bethesda, MD 20892 USA. RP Miotti, PG (reprint author), NIH, Bldg 10, Bethesda, MD 20892 USA. NR 6 TC 1 Z9 1 U1 0 U2 0 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD FEB 23 PY 2000 VL 283 IS 8 BP 999 EP 1000 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 284AM UT WOS:000085308600016 ER PT J AU Ishii, Y Tycko, R AF Ishii, Y Tycko, R TI Multidimensional heteronuclear correlation spectroscopy of a uniformly N-15- and C-13-labeled peptide crystal: Toward spectral resolution, assignment, and structure determination of oriented molecules in solid-state NMR SO JOURNAL OF THE AMERICAN CHEMICAL SOCIETY LA English DT Article ID NUCLEAR-MAGNETIC-RESONANCE; ANGLE-SPINNING NMR; CHEMICAL-SHIFT TENSORS; DIPOLAR-COUPLING FREQUENCIES; ANISOTROPY CORRELATION NMR; ALANYL-GLYCYL-GLYCINE; MULTIPLE-PULSE NMR; MAGIC-ANGLE; ROTATING SOLIDS; PHOSPHOLIPID-BILAYERS AB New one-, two-, and three-dimensional solid-state NMR spectroscopic methods designed for structural studies of uniformly N-15- and C-13-labeled peptides and proteins in oriented samples are described. These methods provide a means of obtaining resolved spectra, sequential resonance assignments, and structural constraints. Experimental results for model single-crystal peptides and amino acids demonstrate that high-resolution one-dimensional C-13 spectra can be obtained for signals from carbonyl or carboxyl ((CO)-C-13) carbons in uniformly labeled samples by applying phase-modulated selective homonuclear (PSH) decoupling at aliphatic carbon resonances, in addition to heteronuclear proton and N-15 decoupling. C-13-detected two-dimensional N-15/C-13 chemical shift correlation spectroscopy is made possible by a combination of PSH decoupling and broadband heteronuclear polarization transfer sequences such as WALTZ-5 cross-polarization. Experimental two-dimensional spectra of uniformly N-15- and C-13-labeled AlaGlyGly crystals show that resolution and sequential assignment of (CO)-C-13 and N-15 NMR signals is possible. Comparisons of experimental spectra and simulations verify the assignments and the accuracy of structural information contained in the two-dimensional spectra in the form of the orientation-dependent (CO)-C-13 and N-15 chemical shifts. C-13-detected three-dimensional spectroscopy is also demonstrated by adding a H-1-N-15 dipolar dimension to the two-dimensional methods. Results of experiments at fields of 9.39 and 17.6 T (400 and 750 MHz proton NMR frequencies) are reported. Motivations for uniform labeling and C-13 detection in oriented systems and implications for future structural studies of oriented proteins are discussed. C1 NIDDKD, Phys Chem Lab, NIH, Bethesda, MD 20892 USA. RP Tycko, R (reprint author), NIDDKD, Phys Chem Lab, NIH, Bethesda, MD 20892 USA. RI Ishii, Yoshitaka/F-4558-2014 OI Ishii, Yoshitaka/0000-0002-7724-6469 NR 93 TC 31 Z9 31 U1 1 U2 8 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0002-7863 J9 J AM CHEM SOC JI J. Am. Chem. Soc. PD FEB 23 PY 2000 VL 122 IS 7 BP 1443 EP 1455 DI 10.1021/ja9915753 PG 13 WC Chemistry, Multidisciplinary SC Chemistry GA 289TZ UT WOS:000085639200023 ER PT J AU Tcherkasskaya, O Ptitsyn, OB Knutson, JR AF Tcherkasskaya, O Ptitsyn, OB Knutson, JR TI Nanosecond dynamics of tryptophans in different conformational states of apomyoglobin proteins SO BIOCHEMISTRY LA English DT Article ID TIME-RESOLVED FLUORESCENCE; ANISOTROPY DECAY; MOLTEN GLOBULE; STRUCTURAL CHARACTERIZATION; ENERGY-TRANSFER; SPECTRA; DEPOLARIZATION; RESIDUES; DOMAIN; MACROMOLECULES AB Fluorescence anisotropy kinetics were employed to quantify the nanosecond mobility of tryptophan residues in different conformational states (native, molten globule, unfolded) of apomyoglobins. Of particular interest is the similarity between the fluorescence anisotropy decays of tryptophans in the native and molten globule states. We find that, in these compact states, tryptophan residues rotate rapidly within a cone of semiangle 22-25 degrees and a correlation time of 0.5 ns, in addition to rotating together with the whole protein with a correlation time of 7-11 ns. The similar nanosecond dynamics of tryptophan residues in both states suggests that the conformation changes that distinguish the molten globule and native states of apomyoglobins originate from either subtle, slow rearrangements or fast changes distant from these tryptophans. C1 NCI, Lab Expt & Computat Biol, NIH, Bethesda, MD 20892 USA. NHLBI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. RP Tcherkasskaya, O (reprint author), NCI, Lab Expt & Computat Biol, NIH, Bethesda, MD 20892 USA. NR 57 TC 35 Z9 35 U1 1 U2 3 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD FEB 22 PY 2000 VL 39 IS 7 BP 1879 EP 1889 DI 10.1021/bi992117+ PG 11 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 285NY UT WOS:000085396200034 PM 10677239 ER PT J AU Weber, HC Jensen, RT Battey, JF AF Weber, HC Jensen, RT Battey, JF TI Molecular organization of the mouse gastrin-releasing peptide receptor gene and its promoter SO GENE LA English DT Article DE bombesin; gene regulation; gene structure; receptor gene ID BOMBESIN-LIKE PEPTIDES; LUNG-CARCINOMA CELLS; CYCLIC-AMP; TRANSCRIPTION FACTORS; RESPONSE ELEMENT; MESSENGER-RNA; X-CHROMOSOME; AR42J CELLS; EXPRESSION; CLONING AB The murine gastrin-releasing peptide receptor (mGRP-R) is a member of the G protein-coupled receptor family and mediates important physiological actions of its specific ligand, the gastrointestinal hormone/neurotransmitter GRP, including mitogenic properties in the mouse Swiss 3T3 fibroblasts. Glucocorticoids and increases in intracellular cAMP are reported to alter GRP-R gene transcription, but the molecular basis for these effects is unknown. To begin to identify possible gene regulatory mechanisms that are responsible for modifying mGRP-R expression, we determined its structure and investigated its basal promoter activity. We isolated and characterized genomic bacteriophage P1 clones encoding the mouse gastrin-releasing peptide receptor (mGRP-R). By DNA sequencing and Southern blot analyses, we determined the protein coding region to be contained in three exons interrupted by two introns 20 and 2 kb in length. The open reading frame of the putative GRP-R gene encodes for a 384-amino acid protein which demonstrates 48% identity with the mouse BRS-3 protein and 53% identity with the mouse NMB-R protein. The mGRP-R gene locus extends over 29 kb and was mapped to the X-chromosome (DXMit20) utilizing a minisatellite polymorphism in the 5' UTR and by fluorescent in-situ hybridization(FISH). In Swiss 3T3 cells, which natively express mGRP-R, two gene-specific mRNA species of 3 and 7 kb can be detected by Northern blot analysis. With RNase protection assays, and independently with inverse PCR of 5' RACE clones, common mRNA initiation sites were identified clustered between 21 and 61 bp downstream of a TTTAAA motif, which is located 450 bp upstream of the ATG translation start site. However, different polyadenylation sites are utilized. A 2 kb genomic DNA fragment extending from 2147 to 141 bases 5' to the ATG translation start was cloned into a luciferase reporter plasmid and shown to contain promoter activity in Swiss 3T3 and COS-7 cells. Progressive promoter truncations and mutations of a cyclic AMP response element (CRE) located 83 bp upstream of the TTTAAA motif demonstrate that transcriptional mCRP-R activation in Swiss 3T3 cells only occurs when both the TTTAAA motif and the intact CRE site are retained. With the availability of the full structure of the mGRP-R gene and the minimal promoter sequences reported in this study, it will be possible in future studies to investigate the molecular basis for transcriptional regulation of the mGRP-R gene by glucocorticoids, cAMP and other factors. Published by Elsevier Science B.V. All rights reserved. C1 NIDDKD, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. Natl Inst Deafness & Other Commun Disorders, NIH, Bethesda, MD USA. RP Jensen, RT (reprint author), NIDDKD, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. NR 39 TC 5 Z9 5 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 FEB 22 PY 2000 VL 244 IS 1-2 BP 137 EP 149 DI 10.1016/S0378-1119(99)00563-6 PG 13 WC Genetics & Heredity SC Genetics & Heredity GA 290DJ UT WOS:000085660700016 PM 10689196 ER PT J AU Valentine, G Chakravarty, S Sarvey, J Bramham, C Herkenham, M AF Valentine, G Chakravarty, S Sarvey, J Bramham, C Herkenham, M TI Fragile X (fmr1) mRNA expression is differentially regulated in two adult models of activity-dependent gene expression SO MOLECULAR BRAIN RESEARCH LA English DT Article DE long-term potentiation; LTP; hippocampus; learning; mental retardation; electroconvulsive shock; immediate-early genes; in situ hybridization ID MENTAL-RETARDATION PROTEIN; RECEPTOR MESSENGER-RNA; LONG-TERM POTENTIATION; IMMEDIATE-EARLY GENE; RAT-BRAIN; MEMORY STORAGE; PERFORANT PATH; KNOCKOUT MICE; DENDRITES; LTP AB We sought to determine whether the fragile X mental retardation gene fmr1 is regulated in long-term potentiation (LTP) and electroconvulsive shock (ECS). in situ hybridization of fmr1 mRNA in hippocampus of rats given LTP in vivo showed no change in fmr1 mRNA levels relative to control. However, ECS induced a selective increase in fmr1 mRNA expression in the dentate gyrus (DG) granule cell layer at 6 h post-ECS. The ECS paradigm may unmask relevant activity-dependent regulatory mechanisms that modulate fmr1 gene transcription in vivo. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIMH, Funct Neuroanat Sect, Bethesda, MD 20892 USA. Uniformed Serv Univ Hlth Sci, Dept Pharmacol, Bethesda, MD 20814 USA. Univ Bergen, Preclin Inst, Dept Physiol, Bergen, Norway. RP Herkenham, M (reprint author), NIMH, Funct Neuroanat Sect, Bldg 36,Rm 2D15, Bethesda, MD 20892 USA. OI Herkenham, Miles/0000-0003-2228-4238; Bramham, Clive/0000-0001-5958-7115 NR 32 TC 11 Z9 11 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0169-328X J9 MOL BRAIN RES JI Mol. Brain Res. PD FEB 22 PY 2000 VL 75 IS 2 BP 337 EP 341 DI 10.1016/S0169-328X(99)00310-1 PG 5 WC Neurosciences SC Neurosciences & Neurology GA 288LN UT WOS:000085564400019 ER PT J AU D'Agata, V Zhao, WQ Cavallaro, S AF D'Agata, V Zhao, WQ Cavallaro, S TI Cloning and distribution of the rat parkin mRNA SO MOLECULAR BRAIN RESEARCH LA English DT Article DE autosomal recessive juvenile parkinsonism; parkin; rat; distribution ID RECESSIVE JUVENILE PARKINSONISM; HOMOZYGOUS DELETIONS; JAPANESE FAMILIES; EARLY-ONSET; GENE; MUTATIONS AB We have isolated by RT-PCR and sequenced a partial cDNA coding for the rat homolog of parkin, a gene mutated in autosomal recessive juvenile parkinsonism. The 1.46 kb rat cDNA clone contains a 1376 bp coding sequence that shares strong similarity with the human parkin cDNA. RT-PCR and in situ hybridization revealed widespread expression of parkin in the rat brain and the periphery. The availability of the rat parkin cDNA and the initial elucidation of its distribution should facilitate further research on the pathophysiological role of parkin in the nervous system. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Italian Res Council, Inst Bioimaging & Pathophysiol Cent Nervous Syst, I-95123 Catania, Italy. IRCCS, Oasi Inst Res Mental Retardat & Brain Aging, Troina, EN, Italy. NINDS, Lab Adaptive Syst, NIH, Bethesda, MD 20892 USA. RP Cavallaro, S (reprint author), CNR, Ist Bioimmagini & Fisiopatol Sistema Nervoso Cent, Piazza Roma 2, I-95123 Catania, Italy. RI D'Agata, Velia/F-3622-2010; Cavallaro, Sebastiano/F-3104-2010 OI D'Agata, Velia/0000-0003-1114-8265; Cavallaro, Sebastiano/0000-0001-7590-1792 NR 13 TC 24 Z9 25 U1 1 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0169-328X J9 MOL BRAIN RES JI Mol. Brain Res. PD FEB 22 PY 2000 VL 75 IS 2 BP 345 EP 349 DI 10.1016/S0169-328X(99)00286-7 PG 5 WC Neurosciences SC Neurosciences & Neurology GA 288LN UT WOS:000085564400021 ER PT J AU Frattali, CM Grafman, J Patronas, N Makhlouf, F Litvan, I AF Frattali, CM Grafman, J Patronas, N Makhlouf, F Litvan, I TI Language disturbances in corticobasal degeneration SO NEUROLOGY LA English DT Article DE corticobasal degeneration; aphasia; language disturbances AB To characterize the language deficits in corticobasal degeneration (CBD) and their relation to neuroradiologic findings, the authors administered a standardized battery of neurobehavioral tests and performed MRI studies on 15 patients with CBD. Eight patients (53%) had classifiable aphasias, including anemic, Broca's and transcortical motor aphasias. The aphasias were associated primarily with left frontal and parietal cortical damage and subcortical white matter and corpus callosum abnormalities. Our findings demonstrate that language disturbances in CBD are more frequent than previously considered. C1 Henry M Jackson Fdn, Cognit Neuropharmacol Unit, Bethesda, MD 20817 USA. NIH, Speech Language Pathol Sect, Dept Rehabil Med, Ctr Clin, Bethesda, MD 20892 USA. NIH, Dept Radiol, Ctr Clin, Bethesda, MD 20892 USA. NINDS, Cognit Neurosci Sect, Bethesda, MD 20892 USA. NIH, Ctr Informat & Technol, Bethesda, MD 20892 USA. RP Litvan, I (reprint author), Henry M Jackson Fdn, Cognit Neuropharmacol Unit, Champlain Bldg,6410 Rockledge Dr,Suite 600, Bethesda, MD 20817 USA. OI Grafman, Jordan H./0000-0001-8645-4457; Litvan, Irene/0000-0002-3485-3445 NR 10 TC 48 Z9 50 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 FEB 22 PY 2000 VL 54 IS 4 BP 990 EP 992 PG 3 WC Clinical Neurology SC Neurosciences & Neurology GA 284ZL UT WOS:000085362700043 PM 10691002 ER PT J AU Kitade, Y Wakana, M Tsuboi, T Yatome, C Bayly, SF Player, MR Torrence, PF AF Kitade, Y Wakana, M Tsuboi, T Yatome, C Bayly, SF Player, MR Torrence, PF TI 2-methyladenosine-substituted 2 ',5 '-oligoadenylates: Conformations, 2-5A binding and catalytic activities with human ribonuclease L SO BIOORGANIC & MEDICINAL CHEMISTRY LETTERS LA English DT Article ID BIOLOGICAL-ACTIVITIES; ACTIVATION ABILITIES; ANALOGS; ENZYME AB 2-Methyladenosine-substituted analogues of 2-5A, p5'A2'p5'A2'p5'(me(2)A), p5'(me(2)A)2'p5'A2'p5'A, and p5'(me(2)A) 2'p5'(me(2)A)2'p5'(me(2)A), were prepared via a modification of a lead ion-catalyzed ligation reaction. These 5'-monophosphates were subsequently converted into the corresponding 5'-triphosphates. Both binding and activation of human recombinant RNase L by various 2-methyladenosine-substituted 2-5A analogues were examined. Among the 2-5A analogues, p5'A2'p5'A2'p5'(me(2)A) showed the strongest binding affinity and was as effective as 2-5A itself as an activator of RNase L. The CD spectra of both p5'(me(2)A)2'p5'A2'p5'A and p5'A2'p5'A2'p5'(me(2)A) were superimposable on that of p5'A2'p5'A2'p5'A. indicative of an anti orientation about the base-glycoside bonds as in naturally occurring 2-5A. (C) 2000 Elsevier Science Ltd. All rights reserved. C1 Gifu Univ, Fac Engn, Dept Biomol Sci, Lab Mol Biochem, Gifu 5011193, Japan. NIDDKD, Sect Biomed Chem, Med Chem Lab, NIH, Bethesda, MD 20892 USA. No Arizona Univ, Dept Chem, Flagstaff, AZ 86011 USA. RP Kitade, Y (reprint author), Gifu Univ, Fac Engn, Dept Biomol Sci, Lab Mol Biochem, Gifu 5011193, Japan. NR 14 TC 6 Z9 6 U1 0 U2 6 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0960-894X J9 BIOORG MED CHEM LETT JI Bioorg. Med. Chem. Lett. PD FEB 21 PY 2000 VL 10 IS 4 BP 329 EP 331 DI 10.1016/S0960-894X(99)00703-9 PG 3 WC Chemistry, Medicinal; Chemistry, Organic SC Pharmacology & Pharmacy; Chemistry GA 288AJ UT WOS:000085539400005 PM 10714492 ER PT J AU Blair, PJ Riley, JL Harlan, DM Abe, R Tadaki, DK Hoffmann, SC White, L Francomano, T Perfetto, SJ Kirk, AD June, CH AF Blair, PJ Riley, JL Harlan, DM Abe, R Tadaki, DK Hoffmann, SC White, L Francomano, T Perfetto, SJ Kirk, AD June, CH TI CD40 ligand (CD154) triggers a short-term CD4(+) T cell activation response that results in secretion of immunomodulatory cytokines and apoptosis SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Article DE costimulatory molecules; T lymphocytes; transplantation; cytokines; apoptosis ID CO-STIMULATION; CYCLOSPORINE-A; ALLOGRAFT-REJECTION; SIGNALING MOLECULE; CROSS-LINKING; LYMPHOCYTES; CD28; EXPRESSION; INDUCTION; MICE AB Signals generated through CD28-B7 and CD40 Ligand (CD40L)-CD40 interactions have been shown to be crucial for the induction of long-term allograft survivability. We have recently demonstrated that humanized anti-CD40L (hu5C8) prevents rejection of mismatched renal allografts in primates. To investigate potential mechanisms of CD40L-induced allograft acceptance, we coimmobilized hu5C8 with suboptimal amounts of anti-CD3 to stimulate CD4(+) T cells. We now report that anti-CD3/CD40L costimulation results in CD28-independent activation and subsequent deletion of resting T cells. Coligation of CD3 and CD40L increased expression of CD69, CD25, and CD54 on CD4(+) T cells. We also round that costimulation with anti-CD3/CD40L resulted in enhanced production of interleukin (IL)-10, interferon gamma, and turner necrosis factor alpha but not IL-2 or IL-6. Interestingly, alter several days, anti-CD3/CD40L-mediated activation was followed by apoptosis in a significant population of cells. Consistent with that observation, anti-CD3/CD40L did not enhance the antiapoptotic proteins Bcl-2 and Bcl-xL. Further, the addition of CD28 at 24 h failed to rescue those cells induced to dir after costimulation with anti-CD3/CD40L. Together, these data suggest that the graft-sparing effect of hu5C8 in vivo may result in part from early and direct effects on CD4(+) T cells, including a vigorous induction of immunomodulatory cytokines and/or apoptosis of allograft-specific T cells. C1 NIDDK, Natl Naval Med Ctr, Naval Transplantat & Autoimmun Branch 034, Bethesda, MD 20889 USA. Univ Penn, Philadelphia, PA 19104 USA. Sci Univ Tokyo, Res Inst Biol Sci, Noda, Chiba 2780022, Japan. Henry M Jackson Fdn Advancement Mil Med, Bethesda, MD 20889 USA. RP Blair, PJ (reprint author), NIDDK, Natl Naval Med Ctr, Naval Transplantat & Autoimmun Branch 034, 8901 Wisconsin Ave, Bethesda, MD 20889 USA. RI Kirk, Allan/B-6905-2012; OI Riley, James/0000-0002-1057-576X NR 52 TC 133 Z9 138 U1 0 U2 1 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 FEB 21 PY 2000 VL 191 IS 4 BP 651 EP 660 DI 10.1084/jem.191.4.651 PG 10 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 287MK UT WOS:000085509600008 PM 10684857 ER PT J AU Champion, JM Kean, RB Rupprecht, CE Notkins, AL Koprowski, H Dietzschold, B Hooper, DC AF Champion, JM Kean, RB Rupprecht, CE Notkins, AL Koprowski, H Dietzschold, B Hooper, DC TI The development of monoclonal human rabies virus-neutralizing antibodies as a substitute for pooled human immune globulin in the prophylactic treatment of rabies virus exposure SO JOURNAL OF IMMUNOLOGICAL METHODS LA English DT Article DE rabies virus; human monoclonal antibodies; rabies post-exposure prophylaxis; passive immunization ID CENTRAL-NERVOUS-SYSTEM; EPSTEIN-BARR VIRUS; POSTEXPOSURE TREATMENT; GLYCOPROTEIN; LYMPHOCYTES; CLEARANCE; RESPONSES; PROTECTS; ANTIGENS AB To provide a more defined and safer replacement for the human rabies immune globulin (HRIG) from pooled serum which is currently used for treatment of exposure to rabies virus we have developed a series of human rabies virus-specific monoclonal antibodies. Mouse-human heterohybrid myeloma cells producing rabies virus-specific human monoclonal antibodies were prepared using B cells obtained from volunteers recently-immunized with a commercial rabies virus vaccine (HDCV). Cell lines producing antibody which neutralized the Evelyn-Rokitnicki-Abelseth (ERA) rabies virus strain in vitro were cloned and the resulting monoclonal antibodies characterized for isotype, specificity against a variety of rabies virus isolates, and neutralization capacity. The ability of the monoclonal antibodies to neutralize a variety of rabies virus strains in vitro correlated with their binding specificity for these viruses in an enzyme-linked immunoadsorbant assay (ELISA), A number of these antibodies have proven suitable for the formulation of a prophylactic human monoclonal antibody-based reagent which would provide significant advantages to the HRIG in having defined, reproducible specificity, lessened possibility of contamination with viral pathogens, and consistent availability. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Thomas Jefferson Univ, Dept Microbiol & Immunol, Ctr Neurovirol, Philadelphia, PA 19107 USA. Ctr Dis Control & Prevent, Rabies Sect, Atlanta, GA 30333 USA. NIDR, Oral Med Lab, NIH, Bethesda, MD 20892 USA. RP Hooper, DC (reprint author), Thomas Jefferson Univ, Dept Microbiol & Immunol, Ctr Neurovirol, Rm 454 JAH,1020 Locust St, Philadelphia, PA 19107 USA. OI Hooper, Douglas/0000-0002-8578-5104 FU NIDCR NIH HHS [N01-DE52606] NR 29 TC 33 Z9 41 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0022-1759 J9 J IMMUNOL METHODS JI J. Immunol. Methods PD FEB 21 PY 2000 VL 235 IS 1-2 BP 81 EP 90 DI 10.1016/S0022-1759(99)00223-9 PG 10 WC Biochemical Research Methods; Immunology SC Biochemistry & Molecular Biology; Immunology GA 290JY UT WOS:000085674200009 PM 10675760 ER PT J AU Hennighausen, L AF Hennighausen, L TI Mouse models for breast cancer SO ONCOGENE LA English DT Article DE mammary; cancer; mouse models; transgenic; knock-out ID TRANSGENIC MICE; GENES C1 NIDDKD, Lab Genet & Physiol, NIH, Bethesda, MD 20892 USA. RP Hennighausen, L (reprint author), NIDDKD, Lab Genet & Physiol, NIH, Bldg 8,Room 101, Bethesda, MD 20892 USA. NR 4 TC 20 Z9 21 U1 0 U2 0 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD FEB 21 PY 2000 VL 19 IS 8 SI SI BP 966 EP 967 DI 10.1038/sj.onc.1203346 PG 2 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 292LT UT WOS:000085796400001 PM 10713679 ER PT J AU Cardiff, RD Anver, MR Gusterson, BA Hennighausen, L Jensen, RA Merino, MJ Rehm, S Russo, J Tavassoli, FA Wakefield, LM Ward, JM Green, JE AF Cardiff, RD Anver, MR Gusterson, BA Hennighausen, L Jensen, RA Merino, MJ Rehm, S Russo, J Tavassoli, FA Wakefield, LM Ward, JM Green, JE TI The mammary pathology of genetically engineered mice: the consensus report and recommendations from the Annapolis meeting SO ONCOGENE LA English DT Article DE genetically engineered mice; mouse mammary models; breast tumor classifications ID MYC-INDUCED TUMORIGENESIS; SCARFF-BLOOM-RICHARDSON; ALPHA TRANSGENIC MICE; TRUNCATED INT3 GENE; GROWTH-FACTOR-ALPHA; TGF-ALPHA; HISTOLOGICAL GRADES; METASTATIC DISEASE; BREAST CARCINOMAS; GLAND INVOLUTION AB NIH sponsored a meeting of medical and veterinary pathologists with mammary gland expertise in Annapolis in March 1999, Rapid development of mouse mammary models has accentuated the need for definitions of the mammary lesions in genetically engineered mice (GEM) and to assess their usefulness as models of human breast disease. The panel of nine pathologists independently reviewed material representing over 90% of the published systems, The GEM tumors were found to have: (1) phenotypes similar to those of non-GEM; (2) signature phenotypes specific to the transgene; and (3) some morphological similarities to the human disease. The current mouse mammary and human breast tumor classifications describe the majority of GEM lesions but unique morphologic lesions are found in many GEM. Since little information is available on the natural history of GEM lesions, a simple morphologic nomenclature is proposed that allows direct comparisons between models. Future progress requires rigorous application of guidelines covering pathologic examination of the mammary gland and the whole animal. Since the phenotype of the lesions is an essential component of their molecular pathology, funding agencies should adopt policies ensuring careful morphological evaluation of any funded research involving animal models. A pathologist should be part of each research team. C1 Univ Calif Davis, UCD Ctr Comparat Med, Davis, CA 95616 USA. NCI, Pathol Histotechnol Lab, SAIC Frederick, FCRDC, Frederick, MD 21702 USA. Inst Canc Res, Breakthrough toby Robins Breast Canc Res Ctr, London SW3 6JB, England. NIDDK, Lab Genet & Physiol, NIH, Bethesda, MD 20892 USA. Vanderbilt Univ, Med Ctr, Dept Pathol, Nashville, TN 37232 USA. NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. Fox Chase Canc Ctr, Breast Canc Res Lab, Philadelphia, PA 19111 USA. SmithKline Beecham Pharmaceut, King Of Prussia, PA 19406 USA. Armed Forces Inst Pathol, Dept Gynecol & Breast Pathol, Washington, DC 20306 USA. NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bethesda, MD 20892 USA. NCI, Vet & Tumor Pathol Sect, Off Lab Anim Resources, Frederick, MD 21702 USA. RP Cardiff, RD (reprint author), Univ Calif Davis, UCD Ctr Comparat Med, Cty Rd 98 & Hutchison Dr, Davis, CA 95616 USA. RI gusterson, barry/D-3752-2009; Jensen, Roy/B-9739-2011 OI Jensen, Roy/0000-0003-4430-2281 FU NCI NIH HHS [N01-CO-56000] NR 68 TC 316 Z9 322 U1 0 U2 3 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD FEB 21 PY 2000 VL 19 IS 8 SI SI BP 968 EP 988 DI 10.1038/sj.onc.1203277 PG 21 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 292LT UT WOS:000085796400002 PM 10713680 ER PT J AU Evans, JA Wagner, U Santos, CM Hennighausen, L AF Evans, JA Wagner, U Santos, CM Hennighausen, L TI The interactive web-based histology atlas system SO ONCOGENE LA English DT Article DE histology; internet; breast; carcinogenesis; mouse models AB Molecular pathways and mechanisms underlying human cancer are frequently investigated in animal models. Studies to identify and dissect molecular pathways include the discovery of genes and their subsequent analysis in transgenic and 'knock-out' mice. A critical aspect in such investigations is the evaluation of organ integrity and histology upon the alteration or inactivation of specific genes. Results from such studies are usually published in scientific journals. However, due to print space and costs the display of large high quality images is limited, Furthermore, the printed media does not permit an easy comparison of histological images published in different journals and different years; The Internet provides a tool for the timely and inexpensive dissemination of scientific data to the research community. However, its potential for the analysis of histological images has not been explored. Here we present a web-based interactive histology atlas (http://histology,nih,gov) that permits the retrieval of annotated, high-resolution histology images via the Internet, This histology atlas also takes advantage of the interactive nature of the Internet to support the communication between different research groups. As an outline forum this atlas provides the framework to evaluate and understand cancer pathology, and to develop a consensus between veterinary and human pathologists. C1 NIH, Ctr Informat Technol, Div Computat Biosci, Bethesda, MD 20892 USA. NIDDKD, Lab Genet & Physiol, NIH, Bethesda, MD 20892 USA. RP Evans, JA (reprint author), NIH, Ctr Informat Technol, Div Computat Biosci, Bldg 12A,Room 1018, Bethesda, MD 20892 USA. NR 0 TC 7 Z9 8 U1 0 U2 1 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD FEB 21 PY 2000 VL 19 IS 8 SI SI BP 989 EP 991 DI 10.1038/sj.onc.1203279 PG 3 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 292LT UT WOS:000085796400003 PM 10713681 ER PT J AU Callahan, R Smith, GH AF Callahan, R Smith, GH TI MMTV-induced mammary tumorigenesis: gene discovery, progression to malignancy and cellular pathways SO ONCOGENE LA English DT Review DE MMTV; Wnt; Fgf; notch; elF3p48/INT6 ID HUMAN BREAST-CANCER; TUMOR VIRUS-DNA; TRANSGENIC MICE; PROVIRAL INSERTION; EPITHELIAL-CELLS; GROWTH-FACTORS; NOTCH-GENE; IN-VIVO; MOUSE DEVELOPMENT; STEM-CELL AB The study of the mouse mammary tumor virus (MMTV) has provided important insights into the mechanisms of gene transcription regulation by steroid hormones, the mode of action of heritable super antigens and the progressive nature of neoplastic transformation in the mammary gland, Here we describe the current situation with respect to the latter aspect of MMTV biology and the prospects for further advance in our understanding of breast cancer in humans that may be expected from a continued study of MMTV-induced mammary neoplasia, MMTV is a heritable somatic mutagen whose target range is limited, Commonly, the tumorigenic capacity of MMTV is restricted to mammary gland, whereas infection is found in a variety of cell types, In order to replicate, proviral DNA must be inserted into the cell DNA and cell division is required to fix the mutation. Yet only in the mammary epithelium does this lead to neoplastic transformation. This suggests a unique relationship between MMTV and mammary epithelium. In evaluating this relationship, we and others have discovered genes and potential gene pathways that are pertinent in mammary differentiation and neoplasia, In addition, the clonal nature of these progressive events from normal to malignant phenotype has become increasingly clear, The weight of these observations compel us to conclude that mammary neoplasms arise from multipotent mammary epithelial cells through a process of acquired mutations that are reflected in the increasingly malignant nature of the population of progeny produced by these damaged stem cells. C1 NCI, Tumor Immunol & Biol Lab, Bethesda, MD 20892 USA. RP Callahan, R (reprint author), NCI, Tumor Immunol & Biol Lab, Bethesda, MD 20892 USA. NR 123 TC 115 Z9 119 U1 0 U2 5 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD FEB 21 PY 2000 VL 19 IS 8 SI SI BP 992 EP 1001 DI 10.1038/sj.onc.1203276 PG 10 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 292LT UT WOS:000085796400004 PM 10713682 ER PT J AU Li, Y Hively, WP Varmus, HE AF Li, Y Hively, WP Varmus, HE TI Use of MMTV-Wnt-1 transgenic mice for studying the genetic basis of breast cancer SO ONCOGENE LA English DT Review DE Wnt-1; breast cancer; cancer models; mammary; transgenic; MMTV ID MAMMARY-TUMOR VIRUS; PROTO-ONCOGENE INT-1; PROVIRAL INSERTION; ESTROGEN-RECEPTOR; GROWTH-FACTOR; BETA-CATENIN; CHROMOSOMAL INSTABILITY; PROGESTERONE RECEPTORS; CELL-PROLIFERATION; CANDIDATE ONCOGENE AB Wnt-1 was first identified as a protooncogene activated by viral insertion in mouse mammary tumors. Transgenic expression of this gene using a mouse mammary tumor virus LTR enhancer causes extensive ductal hyperplasia early in life and mammary adenocarcinomas in approximately 50% of the female transgenic (TG) mice by 6 months of age. Metastasis to the lung and proximal lymph nodes is rare at the time tumors are detected but frequent after the removal of the primary neoplasm, The potent mitogenic effect mediated by Wnt-1 expression does not require estrogen stimulation; tumors form after an increased latency in estrogen receptor alpha-null mice. Several genetic lesions, including inactivation of p53 and over-expression of Fgf-3, collaborate with Wnt-1 in leading to mammary tumors, but loss of Sky and inactivation of one allele of Rb do not affect the rate of tumor formation in Wnt-1 TG mice. C1 NCI, Div Basic Sci, Bethesda, MD 20892 USA. RP Li, Y (reprint author), NCI, Div Basic Sci, 49 Convent Dr,Bldg 49,Room 4A56, Bethesda, MD 20892 USA. OI Li, Yi/0000-0002-9976-518X NR 106 TC 148 Z9 149 U1 2 U2 11 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD FEB 21 PY 2000 VL 19 IS 8 SI SI BP 1002 EP 1009 DI 10.1038/sj.onc.1203273 PG 8 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 292LT UT WOS:000085796400005 PM 10713683 ER PT J AU Green, JE Shibata, MA Yoshidome, K Liu, ML Jorcyk, C Anver, MR Wigginton, J Wiltrout, R Shibata, E Kaczmarczyk, S Wang, W Liu, ZY Calvo, A Couldrey, C AF Green, JE Shibata, MA Yoshidome, K Liu, ML Jorcyk, C Anver, MR Wigginton, J Wiltrout, R Shibata, E Kaczmarczyk, S Wang, W Liu, ZY Calvo, A Couldrey, C TI The C3(1)/SV40 T-antigen transgenic mouse model of mammary cancer: ductal epithelial cell targeting with multistage progression to carcinoma SO ONCOGENE LA English DT Article DE transgenic mice; C3(1); SV40 T-antigen; mammary cancer; prostate cancer; rat prostate steroid binding protein; tumor progression; DCIS ID TUMOR PROGRESSION; BREAST-CANCER; RESPONSE ELEMENTS; MICE; GENE; VIRUS; SV40; PROSTATE; EXPRESSION; BINDING AB The 5' flanking region of the C3(1) component of the rat prostate steroid binding protein (PSBP) has been used to successfully target the expression of the SV40 large T-antigen (Tag) to the epithelium of both the mammary and prostate glands resulting in models of mammary and prostate cancers which histologically resemble the human diseases. Atypia of the mammary ductal epithelium develops at about 8 weeks of age, progressing to mammary intraepithelial neoplasia (resembling human ductal carcinoma in situ [DCIS]) at about 12 weeks of age with the development of invasive carcinomas at about 16 weeks of age in 100% of female mice. The carcinomas share features to what has been classified in human breast cancer as infiltrating ductal carcinomas. All FVB/N female mice carrying the transgene develop mammary cancer with about a 15% incidence of lung metastases. Approximately 10% of older male mice develop anaplastic mammary carcinomas. Unlike many other transgenic models in which hormones and pregnancy are used to induce a mammary phenotype, C3(1)/Tag mice develop mammary tumors in the mammary epithelium of virgin animals without hormone supplementation or pregnancy. Although mammary tumor development appears hormone-responsive at early stages, invasive carcinomas are hormone-independent, which corresponds to the loss of estrogen receptor-a expression during tumor progression. Molecular and biologic factors related to mammary tumor progression can be studied in this model since lesions evolve over a predictable time course. Genomic alterations have been identified during tumor progression, including an amplification of the distal portion of chromosome 6 containing ki-ras and loss of heterozygosity (LOH) in other chromosomal regions. We have demonstrated that stage specific alterations in the expression of genes which are critical regulators of the cell cycle and apoptosis are functionally important in vivo. C3(1)/Tag mice appear useful for testing particular therapies since growth of the mammary tumors can be reduced using chemopreventive agents, cytokines, and an anti-angiogenesis agent. C1 NCI, Transgen Oncogenesis Grp, Lab Cell Regulat & Carcinogneesis, Bethesda, MD 20892 USA. Boise State Univ, Dept Biol, Boise, ID 83725 USA. SAIC, Pathol Histopathol Lab, Frederick, MD 21712 USA. NCI, Lab Expt Immunol, Frederick, MD 21712 USA. RP Green, JE (reprint author), NCI, Transgen Oncogenesis Grp, Lab Cell Regulat & Carcinogneesis, Bldg 41,Room C629, Bethesda, MD 20892 USA. FU NCI NIH HHS [N01-CO-56000] NR 39 TC 114 Z9 114 U1 0 U2 9 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD FEB 21 PY 2000 VL 19 IS 8 SI SI BP 1020 EP 1027 DI 10.1038/sj.onc.1203280 PG 8 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 292LT UT WOS:000085796400007 PM 10713685 ER PT J AU Deng, CX Scott, F AF Deng, CX Scott, F TI Role of the tumor suppressor gene Brca1 in genetic stability and mammary gland tumor formation SO ONCOGENE LA English DT Article DE Brca1; p53; centrosome; G(2)-M checkpoint; tumorigenesis ID CANCER-SUSCEPTIBILITY GENES; EARLY EMBRYONIC LETHALITY; FAMILIAL BREAST-CANCER; SPLICE VARIANTS BRCA1A; CELL-CYCLE; DNA-DAMAGE; TRANSCRIPTIONAL ACTIVATION; CENTROSOME AMPLIFICATION; NUCLEAR PHOSPHOPROTEIN; MEIOTIC CELLS AB Germline mutations in the tumor suppressor BRCA1 predispose women to breast and ovarian cancers. Current evidence demonstrates that mutations in BRCA1 do not directly result in tumor formation, but instead cause genetic instability, subjecting cells to high risks of malignant transformation, In an animal model in which Brca1 is mutated specifically in mammary epithelium, tumorigenesis occurs in mutant glands at low frequency after a long latency. Notably, introduction of a p53-null allele significantly enhanced mammary gland tumor formation in Brca1 conditional mutant mice. These results are consistent with a model that Brca1 is a caretaker gene, whose absence causes genetic instability and triggers further alterations, including inactivation of tumor suppressor genes and/or activation of oncogenes, leading to tumor formation. C1 NIDDKD, Genet Dev & Dis Branch, NIH, Bethesda, MD 20892 USA. RP Deng, CX (reprint author), NIDDKD, Genet Dev & Dis Branch, NIH, 10-9N105, Bethesda, MD 20892 USA. RI deng, chuxia/N-6713-2016 NR 77 TC 101 Z9 104 U1 1 U2 5 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD FEB 21 PY 2000 VL 19 IS 8 SI SI BP 1059 EP 1064 DI 10.1038/sj.onc.1203269 PG 6 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 292LT UT WOS:000085796400012 PM 10713690 ER PT J AU Humphreys, RC Hennighausen, L AF Humphreys, RC Hennighausen, L TI Transforming growth factor alpha and mouse models of human breast cancer SO ONCOGENE LA English DT Article DE TGF alpha; mammary gland; transformation; cancer; mouse models; transgenic ID MAMMARY EPITHELIAL-CELLS; PROTEIN-KINASE-A; TRANSGENIC MICE; TGF-ALPHA; FACTOR RECEPTOR; C-MYC; NEU PROTOONCOGENE; CYCLIN D1; CONSTITUTIVE ACTIVATION; SYNERGISTIC INTERACTION AB Transforming growth factor alpha (TGF alpha) is a principal molecule in the normal and neoplastic development of the mammary gland. Binding of TGF alpha to the epidermal growth factor receptor (EGFR), activates the EGFRs' endogenous tyrosine kinase activity and stimulates growth of the epithelium in the virgin and pregnant mouse mammary gland. TGF alpha expression can be detected in breast cancer cells in vivo and in vitro and overexpression can elicit partial transformation or immortalized human and rodent mammary epithelial cells. Despite evidence implicating TGF alpha in the development of mammary neoplasia, the actual mechanism of TGF alpha-induced transformation is unclear. Transgenic mouse models targeting heterologous TGF alpha to the mammary gland have established TGF alpha overexpression can induce hyperproliferation, hyperplasia and occasional carcinoma. These transgenic studies demonstrated a facilitating, proliferative role for TGF alpha in the development of neoplasia and implicated several oncogenes that can cooperate with TGF alpha to transform the mammary epithelium. From studies of EGFR signaling pathways, inhibitory and modulating agents such as anti-ECFR antibodies and specific kinases inhibitors have been used to block the action of this pathway and prevent the development of TGF alpha-induced neoplasia and tumor formation, Studies in Stat5a knockout mice have established that the JAK2/Stat5a pathway can facilitate the survival of the mammary epithelium and can impact the progression of TGF alpha-mandated mammary tumorigenesis, Together these experiments indicate that TGF alpha and the EGFR signaling pathway are potentially amenable to therapies for treatment of human breast disease. C1 NIDDKD, NIH, Lab Genet & Physiol, Bethesda, MD 20892 USA. RP Humphreys, RC (reprint author), NIDDKD, NIH, Lab Genet & Physiol, Bldg 8,Room 111, Bethesda, MD 20892 USA. NR 75 TC 55 Z9 56 U1 0 U2 0 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD FEB 21 PY 2000 VL 19 IS 8 SI SI BP 1085 EP 1091 DI 10.1038/sj.onc.1203278 PG 7 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 292LT UT WOS:000085796400016 PM 10713694 ER PT J AU Vodovotz, Y Lucia, MS DeLucca, AM Mitchell, JB Kopp, JB AF Vodovotz, Y Lucia, MS DeLucca, AM Mitchell, JB Kopp, JB TI Reduced hematopoietic function and enhanced radiosensitivity of transforming growth factor-beta 1 transgenic mice SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article DE transforming growth factor-beta 1; radiation; transgenic mice; hematopoiesis; bone marrow ID TUMOR-NECROSIS-FACTOR; FACTOR-BETA; BREAST-CANCER; TGF-BETA; DNA FRAGMENTATION; MAMMARY-GLAND; NITRIC-OXIDE; CELL-DEATH; IN-VIVO; GROWTH-FACTOR-BETA-1 AB The cytokine transforming growth factor-beta 1 (TGF-beta 1) has been implicated in some tissue responses to radiation. Previous studies have demonstrated that exogenous TGF-beta 1 increased the lethality of radiation in mice, but the effects of endogenous TGF-beta 1 have not been investigated. To this end, we examined mice that are transgenic for active TGF-beta 1 (Alb/TGF-beta 1), over-expressed via an albumin promoter in the liver with resultant elevation of circulating levels of this cytokine. Alb/TGF-beta 1 mice subjected to 8 Gy of total body irradiation at 3 or 5 weeks of age experienced significantly higher mortality than wild type age- and sex-matched controls by 1 to 2 weeks after irradiation. Alb/TGF-beta 1 3 weeks of age also succumbed to 2 and 4 Gy of whole-body irradiation, while no mortality was observed in wild type mice. Four-week-old Alb/TGF-beta 1 mice exhibited mild anemia and mild uremia. At one week after whole body irradiation with 2 Gy, 4-week-old Alb/TGF-beta 1 mice had significantly reduced white blood cell counts, hematocrit, and platelet counts. Histopathologically, irradiated Alb/TGF-beta 1 mice exhibited decreased bone marrow cellularity and decreased splenic extramedullary hematopoiesis. These results suggest that chronic over-expression of active TGF-beta 1 is associated with increased radiosensitivity and that this effect may be mediated by increased sensitivity of bone marrow to the suppressive effects of radiation. Since TGF-beta 1 levels can be greatly elevated inpatients with certain tumors, these findings may be significant for radiotherapy. Published 2000 Wiley-Liss, Inc.(dagger). C1 NCI, Radiat Biol Branch, Bethesda, MD 20892 USA. Univ Colorado, Hlth Sci Ctr, Dept Pathol, Denver, CO 80262 USA. NIDDKD, Kidney Dis Sect, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. RP Vodovotz, Y (reprint author), Univ Pittsburgh, Sch Med, Dept Surg, 15th Floor Biomed Sci Tower,200 Lothrop St, Pittsburgh, PA 15213 USA. NR 42 TC 12 Z9 13 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD FEB 20 PY 2000 VL 90 IS 1 BP 13 EP 21 DI 10.1002/(SICI)1097-0215(20000220)90:1<13::AID-IJC2>3.0.CO;2-U PG 9 WC Oncology SC Oncology GA 289UJ UT WOS:000085640100002 PM 10725853 ER PT J AU Murray, HW Pepin, J Nutman, TB Hoffman, SL Mahmoud, AAF AF Murray, HW Pepin, J Nutman, TB Hoffman, SL Mahmoud, AAF TI Tropical medicine SO BRITISH MEDICAL JOURNAL LA English DT Review ID CUTANEOUS LEISHMANIASIS; PLASMODIUM-FALCIPARUM; MALARIA; DIAGNOSIS; INFECTION; VACCINE C1 Cornell Univ, Weill Med Coll, Dept Med, New York, NY 10021 USA. Univ Sherbrooke, Ctr Int Hlth, Sherbrooke, PQ J1H 5N4, Canada. NIH, Helminth Immunol Sect, Bethesda, MD 20892 USA. NIH, Clin Parasitol Unit, Parasit Dis Lab, Bethesda, MD 20892 USA. USN, Med Res Ctr, Rockville, MD 20852 USA. Merck Vaccines, Whitehouse Stn, NJ 08889 USA. RP Murray, HW (reprint author), Cornell Univ, Weill Med Coll, Dept Med, New York, NY 10021 USA. FU NIAID NIH HHS [R56 AI016963, AI 16963, R01 AI016963] NR 35 TC 14 Z9 14 U1 0 U2 0 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 0959-8138 J9 BRIT MED J JI Br. Med. J. PD FEB 19 PY 2000 VL 320 IS 7233 BP 490 EP 494 DI 10.1136/bmj.320.7233.490 PG 5 WC Medicine, General & Internal SC General & Internal Medicine GA 288KP UT WOS:000085562200028 PM 10678866 ER PT J AU Mulshine, JL Henschke, CI AF Mulshine, JL Henschke, CI TI Prospects for lung-cancer screening SO LANCET LA English DT Editorial Material ID DIAGNOSIS; LESIONS C1 NCI, Med Branch, Bethesda, MD 20892 USA. Cornell Univ, Weill Med Coll, Ithaca, NY 14853 USA. New York Presbyterian Hosp, New York, NY USA. RP Mulshine, JL (reprint author), NCI, Med Branch, 8900 Rockville Pike, Bethesda, MD 20892 USA. NR 17 TC 28 Z9 28 U1 0 U2 0 PU LANCET LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0140-6736 J9 LANCET JI Lancet PD FEB 19 PY 2000 VL 355 IS 9204 BP 592 EP 593 DI 10.1016/S0140-6736(99)00398-0 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 289HX UT WOS:000085615400006 PM 10696976 ER PT J AU Hatzakis, A Touloumi, G Karanicolas, R Karafoulidou, A Mandalaki, T Anastassopoulou, C Zhang, L Goedert, JJ Ho, DD Kostrikis, LG AF Hatzakis, A Touloumi, G Karanicolas, R Karafoulidou, A Mandalaki, T Anastassopoulou, C Zhang, L Goedert, JJ Ho, DD Kostrikis, LG TI Effect of recent thymic emigrants on progression of HIV-1 disease SO LANCET LA English DT Article ID HEMOPHILIA COHORT; NATURAL-HISTORY; T-CELLS; INFECTION; CCR5; ALLELE; PLASMA; AIDS; GENE; SEROCONVERSION AB Background The concentration of T-cell receptor-rearrangement excision DNA circles (TREC) in peripheral-blood T cells is a marker of recent thymic emigrant ap T cells. We studied the predictive ability of measurements of TREC for clinical outcome in HIV-1-infected individuals. Methods We measured TREC in peripheral-blood mononuclear cells with a real-time PCR assay. We studied 131 Greek participants in the Multicenter Hemophilia Cohort Study who had known HIV-1 seroconversion dates. The prognostic value of baseline TREC, CD4 T-cell count, and HIV-1 RNA concentration was assessed by Kaplan-Meier and Cox's regression analysis. Findings Four participants had progressed to AIDS by first blood sampling. Among the remaining 127 individuals, the median value of TREC per 10(6) cells was 6900 (IQR 2370-15 604). Baseline TREC values were lower in the 53 who progressed to AIDS than in those who did not (geometric mean 2843 [95% CI 1468-5504] vs 6560 [4723-9113] per 10(6) cells; p=0.017). The relative hazard of AIDS, adjusted for plasma viral load, CD4 T-cell count, and age at seroconversion was 1.44 (95% CI 1.04-2.01; p=0.031) per ten-fold increase in TREC; that for death was 1.52 (1.12-2.06; p=0.007). The adjusted relative hazards of death were 2.91 (1.91-4.44; p<0.001) per ten-fold increase in plasma HIV-1 RNA load and 1.20 (1.04-1.38; p=0.014) per 100-cell decrease in CD4 T-cell count. Interpretation The concentration of TREC in the peripheral T-cell pool complements HIV-1 RNA load and CD4 T-cell count in predicting the rate of HIV-1 disease progression. Recent thymic emigrants have a role in the pathogenesis of HIV-1 disease. C1 Rockefeller Univ, Aaron Diamond AIDS Res Ctr, New York, NY 10016 USA. Univ Athens, Sch Med, Dept Hyg & Epidemiol, GR-11527 Athens, Greece. Laikon Hosp, Hemophilia Ctr, Athens, Greece. NCI, Viral Epidemiol Branch, Rockville, MD USA. RP Kostrikis, LG (reprint author), Rockefeller Univ, Aaron Diamond AIDS Res Ctr, 455 1st Ave, New York, NY 10016 USA. RI Kostrikis, Leondios/A-5330-2016 OI Kostrikis, Leondios/0000-0002-5340-7109 FU NIAID NIH HHS [R01 AI40387, AI42848, R01 AI43868] NR 28 TC 77 Z9 81 U1 0 U2 1 PU LANCET LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0140-6736 J9 LANCET JI Lancet PD FEB 19 PY 2000 VL 355 IS 9204 BP 599 EP 604 DI 10.1016/S0140-6736(99)10311-8 PG 6 WC Medicine, General & Internal SC General & Internal Medicine GA 289HX UT WOS:000085615400010 PM 10696979 ER PT J AU Welles, SL Pitt, J Colgrove, R McIntosh, K Chung, PH Colson, A Lockman, S Fowler, MG Hanson, C Landesman, S Moye, J Rich, KC Zorrilla, C Japour, AJ AF Welles, SL Pitt, J Colgrove, R McIntosh, K Chung, PH Colson, A Lockman, S Fowler, MG Hanson, C Landesman, S Moye, J Rich, KC Zorrilla, C Japour, AJ CA Women Infants Transmission Study G TI HIV-1 genotypic zidovudine drug resistance and the risk of maternal-infant transmission in the Women and Infants Transmission Study SO AIDS LA English DT Article DE HIV-1; RT drug-resistant mutations; zidovudine drug-resistance ID HUMAN-IMMUNODEFICIENCY-VIRUS; MOTHER-TO-CHILD; CLINICAL-TRIALS GROUP; HIGH-LEVEL RESISTANCE; TYPE-1 RNA LEVEL; VIRAL LOAD; VERTICAL TRANSMISSION; PERINATAL TRANSMISSION; INFECTION; MUTATIONS AB Objectives: Although the treatment of pregnant women and their infants with zidovudine (ZDV) has been remarkably effective in preventing the perinatal transmission of human HIV-1, many potentially preventable infections still occur. To examine whether the risk of perinatal infection is increased among women who carry ZDV-resistant HIV-1, the role of genotypic ZDV resistance in perinatal transmission was evaluated. Methods: The reverse transcriptase (RT) region of clinical isolates from culture supernatants of 142 HIV-1-infected women enrolled in the Women and Infants Transmission Study (WITS), who had been treated with ZDV during pregnancy was sequenced. Results from genotypic sequencing were linked to demographic, laboratory, and obstetrical databases, and the magnitude of association of having consensus drug-resistant HIV-1 RT mutations with transmission was estimated. Results: Twenty-five per cent (34/142) of maternal isolates had at least one ZDV-associated resistance mutation. A lower CD4 cell percentage and count (P = 0.0001) and higher plasma HIV-1 RNA (P = 0.006) were associated with having any ZDV resistance mutation at delivery. Having any RT resistance mutation [odds ratio (OR): 5.16; 95% confidence interval (CI): 1.40, 18.97; P = 0 0.01], duration of ruptured membranes [OR: 1.13 (1.02, 1.26) per 4 h duration; P = 0.02], and total lymphocyte count [OR: 1.06 (1.01, 1.10) per 50 cells higher revel; P = 0.009] were independently associated with transmission in multivariate analysis. C1 Univ Minnesota, Sch Publ Hlth, Div Epidemiol, Minneapolis, MN 55454 USA. Columbia Univ Coll Phys & Surg, Dept Pediat, Div Infect Dis, New York, NY 10032 USA. Harvard Univ, Sch Med, Beth Israel Deaconess Med Ctr, Div Infect Dis, Boston, MA USA. Harvard Univ, Sch Med, Childrens Hosp, Div Infect Dis, Boston, MA USA. NIH, Div Aids, Bethesda, MD 20892 USA. Texas Childrens Hosp, Baylor Coll Med, Houston, TX 77030 USA. SUNY, Brooklyn, NY USA. NICHHD, Bethesda, MD 20892 USA. Univ Illinois, Dept Pediat, Chicago, IL USA. Univ Puerto Rico, San Juan, PR 00936 USA. RP Welles, SL (reprint author), Univ Minnesota, Sch Publ Hlth, Div Epidemiol, 1300 S 2nd St,Suite 300, Minneapolis, MN 55454 USA. EM welles@epi.umn.edu OI moye, john/0000-0001-9976-8586 FU NIAID NIH HHS [AI139144] NR 42 TC 65 Z9 65 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0269-9370 J9 AIDS JI Aids PD FEB 18 PY 2000 VL 14 IS 3 BP 263 EP 271 DI 10.1097/00002030-200002180-00008 PG 9 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 287HE UT WOS:000085499800008 PM 10716502 ER PT J AU Murphy, E AF Murphy, E TI Mysteries of magnesium homeostasis SO CIRCULATION RESEARCH LA English DT Editorial Material DE heart; beta-adrenergic agonist; glucose transport; insulin; mitochondria ID INTRACELLULAR FREE MAGNESIUM; CYTOSOLIC FREE MAGNESIUM; FREE MG2+ CONCENTRATION; SARCOPLASMIC-RETICULUM; FLUORESCENT INDICATOR; VENTRICULAR MYOCYTES; HEART-CELLS; MG-2+; RAT; TRANSPORT C1 NIEHS, Res Triangle Pk, NC 27709 USA. RP Murphy, E (reprint author), NIEHS, Mail Drop D2-03,Box 12233, Res Triangle Pk, NC 27709 USA. NR 34 TC 62 Z9 65 U1 1 U2 5 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7330 J9 CIRC RES JI Circ.Res. PD FEB 18 PY 2000 VL 86 IS 3 BP 245 EP 248 PG 4 WC Cardiac & Cardiovascular Systems; Hematology; Peripheral Vascular Disease SC Cardiovascular System & Cardiology; Hematology GA 289GX UT WOS:000085613100001 PM 10679471 ER PT J AU Saiardi, A Caffrey, JJ Snyder, SH Shears, SB AF Saiardi, A Caffrey, JJ Snyder, SH Shears, SB TI Inositol polyphosphate multikinase (ArgRIII) determines nuclear mRNA export in Saccharomyces cerevisiae SO FEBS LETTERS LA English DT Article DE ARGRIII; inositol polyphosphate multikinase; inositol phosphate; InsP(6); mRNA export; yeast ID ARGININE METABOLISM; HEXAKISPHOSPHATE; 1,3,4,6-TETRAKISPHOSPHATE; PENTAKISPHOSPHATES; PHOSPHORYLATION; EXPRESSION; TURNOVER; KINASE; GENES AB The ARGRIII gene of Saccharomyces cerevisiae encodes a transcriptional regulator that also has inositol polyphosphate multikinase (ipmk) activity [Saiardi et al, (1999) Curr, Biol, 9, 1323-1326]. To investigate how inositol phosphates regulate gene expression, we disrupted the ARGRIII gene. This mutation impaired nuclear mRNA export, slowed cell growth, increased cellular [InsP(3)] 170-fold and decreased [InsP(6)] 100-fold, indicating reduced phosphorylation of InsP(3) to InsP(6). Levels of diphosphoinositol polyphosphates were decreased much less dramatically than was InsP(6). Low levels of InsP(6), and considerable quantities of Ins(1,3,4,5)P-4, were synthesized by an ipmk-independent route. Transcriptional control by ipmk reflects that it is a pivotal regulator of nuclear mRNA export via inositol phosphate metabolism. (C) 2000 Federation of European Biochemical Societies. C1 NIEHS, Lab Signal Transduct, Inositide Signaling Sect, Res Triangle Pk, NC 27709 USA. Johns Hopkins Univ, Sch Med, Dept Neurosci, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Dept Biochem & Clin Pharmacol, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Dept Psychiat & Behav Sci, Baltimore, MD 21205 USA. RP Shears, SB (reprint author), NIEHS, Lab Signal Transduct, Inositide Signaling Sect, Res Triangle Pk, NC 27709 USA. FU NIDA NIH HHS [DA00074]; NIMH NIH HHS [MH18501] NR 19 TC 101 Z9 103 U1 0 U2 7 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0014-5793 J9 FEBS LETT JI FEBS Lett. PD FEB 18 PY 2000 VL 468 IS 1 BP 28 EP 32 DI 10.1016/S0014-5793(00)01194-7 PG 5 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 286PA UT WOS:000085453300007 PM 10683435 ER PT J AU Yang, XY Wang, LH Chen, TS Hodge, DR Resau, JH DaSilva, L Farrar, WL AF Yang, XY Wang, LH Chen, TS Hodge, DR Resau, JH DaSilva, L Farrar, WL TI Activation of human T lymphocytes is inhibited by peroxisome proliferator-activated receptor gamma (PPAR gamma) agonists - PPAR gamma co-association with transcription factor NFAT SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID EXPRESSION; CELLS; IL-2; GENE AB T lymphocyte activation is highlighted by the induction of interleukin-2 (IL-2) gene expression, which governs much of the early lymphocyte proliferation responses. Peroxisome proliferator-activated receptor-gamma (PPAR gamma) is a member of the nuclear receptor superfamily of ligand-activated transcription factors. PPAR gamma mRNA expression was found in human peripheral blood T lymphocytes, raising the possibility of PPAR gamma involvement in the regulation of T cell function. Here we show that PPAR gamma ligands, troglitazone and 15-deoxy-Delta(12,14) prostaglandin J(2), but not PPAR alpha agonist Wy14643, inhibited IL-2 production and phytohemagglutinin-inducible proliferation in human peripheral blood T-cells in a dose-dependent manner. This inhibitory effect on IL-2 was restricted to the PPAR gamma 2-expressing, not the PPAR gamma-lacking, subpopulation of transfected Jurkat cells. The activated PPAR gamma physically associates with transcriptional factor NFAT regulating the IL-2 promoter, blocking NFAT DNA binding and transcriptional activity. This interaction with T-cell-specific transcription factors indicates an important immunomodulatory role for PPAR gamma in T lymphocytes and could suggest a previously unrecognized clinical potential for PPAR gamma ligands as immunotherapeutic drugs to treat T-cell-mediated diseases by targeting IL-2 gene expression. C1 NCI, Frederick Canc Res & Dev Ctr, Intramural Res Support Program, SAIC Frederick,NIH, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, Cytokine Mol Mechanisms Sect, Lab Mol Immunoregulat,NIH, Frederick, MD 21702 USA. NCI, ABL Basic Res Program, NIH, Frederick, MD 21702 USA. RP Farrar, WL (reprint author), NCI, Frederick Canc Res & Dev Ctr, Intramural Res Support Program, SAIC Frederick,NIH, POB B,Bldg 560,Rm 31-76, Frederick, MD 21702 USA. RI Chen, Taosheng/I-6351-2013 FU NCI NIH HHS [N01-CO-56000] NR 18 TC 268 Z9 285 U1 1 U2 6 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD FEB 18 PY 2000 VL 275 IS 7 BP 4541 EP 4544 DI 10.1074/jbc.275.7.4541 PG 4 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 285FC UT WOS:000085378200002 PM 10671476 ER PT J AU Clifford, GM Londos, C Kraemer, FB Vernon, RG Yeaman, SJ AF Clifford, GM Londos, C Kraemer, FB Vernon, RG Yeaman, SJ TI Translocation of hormone-sensitive lipase and perilipin upon lipolytic stimulation of rat adipocytes SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID DEPENDENT PROTEIN-KINASE; NECROSIS-FACTOR-ALPHA; LIPID DROPLETS; ADENYLATE-CYCLASE; ADIPOSE-TISSUE; LEYDIG-CELLS; INSULIN; MECHANISM; PHOSPHORYLATION; METABOLISM AB Adipocyte lipolysis was compared with hormone-sensitive lipase (HSL)/perilipin subcellular distribution and perilipin phosphorylation using Western blot analysis. Under basal conditions, HSL resided predominantly in the cytosol and unphosphorylated perilipin upon the lipid droplet. Upon lipolytic stimulation of adipocytes isolated from young rats with the beta-adrenergic agonist, isoproterenol, HSL translocated from the cytosol to the lipid droplet, but there was no movement of perilipin from the droplet to the cytosol; however, perilipin phosphorylation was observed. By contrast, upon lipolytic stimulation and perilipin phosphorylation in cells from more mature rats, there was no HSL translocation but a significant movement of perilipin away from the lipid droplet. Adipocytes from younger rats had markedly greater rates of lipolysis than those from the older rats. Thus high rates of lipolysis require translocation of HSL to the lipid droplet and translocation of HSL and perilipin can occur independently of each other. A loss of the ability to translocate HSL to the lipid droplet probably contributes to the diminished lipolytic response to catecholamines with age. C1 Univ Newcastle, Sch Med, Sch Biochem & Genet, Newcastle Upon Tyne NE2 4HH, Tyne & Wear, England. NIDDK, Cellular & Dev Biol Lab, NIH, Bethesda, MD 20892 USA. Vet Affairs Palo Alto Hlth Care Syst, Dept Med, Palo Alto, CA 94304 USA. Hannah Res Inst, Ayr KA6 5HL, Scotland. RP Yeaman, SJ (reprint author), Univ Newcastle, Sch Med, Sch Biochem & Genet, Newcastle Upon Tyne NE2 4HH, Tyne & Wear, England. FU NIDDK NIH HHS [DK46942, DK49705] NR 34 TC 158 Z9 170 U1 0 U2 5 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD FEB 18 PY 2000 VL 275 IS 7 BP 5011 EP 5015 DI 10.1074/jbc.275.7.5011 PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 285FC UT WOS:000085378200067 PM 10671541 ER PT J AU Cheung, AP Struble, E Nguyen, N Liu, P AF Cheung, AP Struble, E Nguyen, N Liu, P TI Stability-indicating high-performance liquid chromatographic assay and analysis of decomposition products of 2-{4-[(7-chloro-2-quinoxalinyl)oxy]phenoxy}propanoic acid SO JOURNAL OF CHROMATOGRAPHY A LA English DT Article; Proceedings Paper CT 23rd International Symposium on High Performance Liquid Phase Separations and Related Techniques CY MAY 30-JUN 04, 1999 CL GRANADA, SPAIN DE pharmaceutical analysis; chloroquinoxalinyloxyphenoxypropanoic acid; aryloxyphenoxypropanoic acid ID CYCLOHEXANEDIONE HERBICIDES; WATER; ARYLOXYPHENOXYPROPIONATE; RESISTANCE AB A stability-indicating HPLC assay has been developed for 2-{4-[(7-chloro-2-quinoxalinyl)oxy]phenoxy}propanic acid (XK469). XK-469 is the 7-chloro analog of herbicide Quizalofop and is currently under development as an antineoplastic agent. HPLC separation of XK469 is achieved with an ODS column using isocratic elution of an aqueous MeOH mobile phase. The assay is reproducible (RSD = 0.9%), linear (r(2) = 0.999), accurate (error = 1.2%) and sensitive (LDL = 1.2 ng). The HPLC separates XK469 from its forced decomposition products. Identities of the decomposition products have been elucidated. (C) 2000 Elsevier Science B.V. All rights reserved. C1 SRI Int, Biopharmaceut Dev Div, Menlo Park, CA 94025 USA. NCI, Pharmaceut Resources Branch, DTP, DCTD,NIH, Bethesda, MD 20892 USA. RP Cheung, AP (reprint author), SRI Int, Biopharmaceut Dev Div, 333 Ravenswood Ave, Menlo Park, CA 94025 USA. FU NCI NIH HHS [N01-CM-77104] NR 15 TC 2 Z9 2 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0021-9673 J9 J CHROMATOGR A JI J. Chromatogr. A PD FEB 18 PY 2000 VL 870 IS 1-2 BP 179 EP 188 DI 10.1016/S0021-9673(99)01093-6 PG 10 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 286QD UT WOS:000085455900021 PM 10722075 ER PT J AU Yang, ZN Mueser, TC Bushman, FD Hyde, CC AF Yang, ZN Mueser, TC Bushman, FD Hyde, CC TI Crystal structure of an active two-domain derivative of Rous sarcoma virus integrase SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE HIV; integrase; protein structure; Rous sarcoma virus; protein X-ray crystallography ID HUMAN-IMMUNODEFICIENCY-VIRUS; RETROVIRAL DNA INTEGRATION; CARBOXYL-TERMINAL DOMAINS; PHOTO-CROSS-LINKING; HIV-1 INTEGRASE; CATALYTIC DOMAIN; BINDING PROPERTIES; PROTEIN INVITRO; NUCLEOSOME CORE; VIRAL-DNA AB Integration of retroviral cDNA is a necessary step in viral replication. The virally encoded integrase protein and DNA sequences at the ends of the linear viral cDNA are required for this reaction. Previous studies revealed that truncated forms of Rous sarcoma virus integrase containing two of the three protein domains can carry out integration reactions in vitro. Here, we describe the crystal structure at 2.5 Angstrom resolution of a fragment of the integrase of Rous sarcoma virus (residues 49-286) containing both the conserved catalytic domain and a modulatory DNA-binding domain (C domain). The catalytic domains form a symmetric dimer, but the C domains associate asymmetrically with each other and together adopt a canted conformation relative to the catalytic domains. A binding path for the viral cDNA is evident spanning both domain surfaces, allowing modeling of the larger integration complexes that are known to be active in vivo. The modeling suggests that formation of an integrase tetramer (a dimer of dimers) is necessary and sufficient for joining both viral cDNA ends at neighboring sites in the target DNA. The observed asymmetric arrangement of C domains suggests that they could form a rotationally symmetric tetramer that may be important for bridging integrase complexes at each cDNA end. C1 NIAMSD, Struct Biol Res Lab, NIH, Bethesda, MD 20892 USA. Salk Inst, Infect Dis Lab, La Jolla, CA USA. RP Hyde, CC (reprint author), NIAMSD, Struct Biol Res Lab, NIH, Bethesda, MD 20892 USA. FU NIAID NIH HHS [AI34786]; NIGMS NIH HHS [GM56553] NR 64 TC 92 Z9 95 U1 0 U2 2 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-2836 J9 J MOL BIOL JI J. Mol. Biol. PD FEB 18 PY 2000 VL 296 IS 2 BP 535 EP 548 DI 10.1006/jmbi.1999.3463 PG 14 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 290VQ UT WOS:000085698900018 PM 10669607 ER PT J AU Mueser, TC Jones, CE Nossal, NG Hyde, CC AF Mueser, TC Jones, CE Nossal, NG Hyde, CC TI Bacteriophage T4 gene 59 helicase assembly protein binds replication fork DNA. The 1.45 angstrom resolution crystal structure reveals a novel alpha-helical two-domain fold SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE helicase; helicase assembly factor; DNA replication; fork-DNA binding; HMG domain ID POLAR BRANCH MIGRATION; ESCHERICHIA-COLI; COMPLEX; ORIGIN; PURIFICATION; DIFFRACTION; MOTIF; HMG1 AB The bacteriophage T4 gene 59 helicase assembly protein is required for recombination-dependent DNA replication, which is the predominant mode of DNA replication in the late stage of T4 infection. T4 gene 59 helicase assembly protein accelerates the loading of the T4 gene 41 helicase during DNA synthesis by the T4 replication system in vitro. T4 gene 59 helicase assembly protein binds to both T4 gene 41 helicase and T4 gene 32 single-stranded DNA binding protein, and to single and double-stranded DNA. We show here that T4 gene 59 helicase assembly protein binds most tightly to fork DNA substrates, with either single or almost entirely double-stranded arms. Our studies suggest that the helicase assembly protein is responsible for loading T4 gene 41 helicase specifically at replication forks, and that its binding sites for each arm must hold more than six, but not more than 12 nucleotides. The 1.45 Angstrom resolution crystal structure of the full-length 217-residue monomeric T4 gene 59 helicase assembly protein reveals a novel cl-helical bundle fold with two domains of similar size. Surface residues are predominantly basic (pI 9.37) with clusters of acidic residues but exposed hydrophobic residues suggest sites for potential contact with DNA and with other protein molecules. The N-terminal domain has structural similarity to the double-stranded DNA binding domain of rat HMG1A. We propose a speculative model of how the T4 gene 59 helicase assembly protein might bind to fork DNA based on the similarity to HMG1, the location of the basic and hydrophobic regions, and the site size of the fork arms needed for tight fork DNA binding. The fork-binding model suggests putative binding sites for the T4 gene 32 single-stranded DNA binding protein and for the hexameric T4 gene 41 helicase assembly. C1 NIAMSD, Struct Biol Res Lab, NIH, Bethesda, MD 20892 USA. NIDDKD, Mol & Cellular Biol Lab, NIH, Bethesda, MD 20892 USA. RP Mueser, TC (reprint author), NIAMSD, Struct Biol Res Lab, NIH, Bldg 6,Room B2-34A, Bethesda, MD 20892 USA. NR 57 TC 38 Z9 38 U1 0 U2 1 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-2836 J9 J MOL BIOL JI J. Mol. Biol. PD FEB 18 PY 2000 VL 296 IS 2 BP 597 EP 612 DI 10.1006/jmbi.1999.3438 PG 16 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 290VQ UT WOS:000085698900022 PM 10669611 ER PT J AU Sieber, SM Galson, SK AF Sieber, SM Galson, SK TI Cooperating on childhood cancer SO SCIENCE LA English DT Letter C1 NCI, NIH, Special Projects, Bethesda, MD 20892 USA. US EPA, Off Sci Coordinat & Policy, Washington, DC 20460 USA. RP Sieber, SM (reprint author), NCI, NIH, Special Projects, 31 Ctr Dr, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 USA SN 0036-8075 J9 SCIENCE JI Science PD FEB 18 PY 2000 VL 287 IS 5456 BP 1205 EP 1205 PG 1 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 286KG UT WOS:000085444700023 PM 10712154 ER PT J AU McNally, JG Muller, WG Walker, D Wolford, R Hager, GL AF McNally, JG Muller, WG Walker, D Wolford, R Hager, GL TI The glucocorticoid receptor: Rapid exchange with regulatory sites in living cells SO SCIENCE LA English DT Article ID TRANSCRIPTION FACTORS; MMTV PROMOTER; IN-VIVO; GENES; ACTIVATION; CHROMATIN; INTERPLAY; INVIVO AB Steroid receptors bind to site-specific response elements in chromatin and modulate gene expression in a hormone-dependent fashion. With the use of a tandem array of mouse mammary tumor virus reporter elements and a form of glucocorticoid receptor labeled with green fluorescent protein, targeting of the receptor to response elements in live mouse cells was observed. Photobleaching experiments provide direct evidence that the hormone-occupied receptor undergoes rapid exchange between chromatin and the nucleoplasmic compartment. Thus, the interaction of regulatory proteins with target sites in chromatin is a more dynamic process than previously believed. C1 NCI, Lab Receptor Biol & Gene Express, Bethesda, MD 20892 USA. RP Hager, GL (reprint author), NCI, Lab Receptor Biol & Gene Express, Bldg 41,Room B602, Bethesda, MD 20892 USA. NR 34 TC 552 Z9 563 U1 0 U2 17 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 USA SN 0036-8075 J9 SCIENCE JI Science PD FEB 18 PY 2000 VL 287 IS 5456 BP 1262 EP 1265 DI 10.1126/science.287.5456.1262 PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 286KG UT WOS:000085444700049 PM 10678832 ER PT J AU Fauci, AS AF Fauci, AS TI AIDS in the 21st century. Reply SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Letter C1 NIH, Bethesda, MD 20892 USA. RP Fauci, AS (reprint author), NIH, Bldg 10, Bethesda, MD 20892 USA. NR 4 TC 0 Z9 0 U1 0 U2 0 PU MASSACHUSETTS MEDICAL SOC PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD FEB 17 PY 2000 VL 342 IS 7 BP 516 EP 517 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 284FA UT WOS:000085319100022 ER PT J AU Bach, PB Cramer, LD Begg, CB Warren, JL AF Bach, PB Cramer, LD Begg, CB Warren, JL TI Racial differences in the treatment of early-stage lung cancer. Reply SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Letter C1 Mem Sloan Kettering Canc Ctr, New York, NY 10021 USA. NCI, Bethesda, MD 20892 USA. RP Bach, PB (reprint author), Mem Sloan Kettering Canc Ctr, 1275 York Ave, New York, NY 10021 USA. NR 5 TC 1 Z9 1 U1 0 U2 1 PU MASSACHUSETTS MEDICAL SOC PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD FEB 17 PY 2000 VL 342 IS 7 BP 518 EP 519 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 284FA UT WOS:000085319100026 ER PT J AU Ivanov, VN Kehrl, JH Ronai, Z AF Ivanov, VN Kehrl, JH Ronai, Z TI Role of TRAF2/GCK in melanoma sensitivity to UV-induced apoptosis SO ONCOGENE LA English DT Article DE TRAF2; GCK; NF-kappa B; melanoma; apoptosis; UV-irradiation ID NF-KAPPA-B; RECEPTOR-ASSOCIATED FACTOR-2; FAS LIGAND EXPRESSION; FACTOR TNF RECEPTOR-1; TERMINAL KINASE; SIGNAL-TRANSDUCTION; INDUCED ACTIVATION; CELL-DEATH; T-CELLS; PATHWAYS AB Radiation resistance is a hallmark of human melanoma, and yet mechanisms underlying this resistance are not well understood. We recently established the role of ATF2 in this process, suggesting that stress kinases, which contribute to regulation of ATF2 stability and activity, play an important role in the acquisition of such resistance. Here we demonstrate that changes in the expression and respective activities of TRAF2/GCK occur during melanoma development and regulate its sensitivity to UV-induced apoptosis, Comparing early- and late-stage melanoma cells revealed low expression of TRAF2 and GCK in early-stage melanoma, which coincided with poor resistance to UV-induced, TNF-mediated apoptosis; forced expression of GCK alone or in combination with TRAF2 efficiently increased JNK and NF-kappa B activities, which coincided with increased protection against apoptosis, Conversely, forced expression of the dominant negative form of TRAF2 or GCK in late-stage melanoma cells reduced NF-kappa B activity and decreased Fas expression, resulting in a lower degree of UV-induced, Fas-mediated cell death. Our results illustrate a mechanism in which protection from, or promotion of, UV-induced melanoma cell death depends on the nature of the apoptotic cascade (TNF or Fas) and on the availability of TRAF2/GCK, whose expression increases during melanoma progression. C1 CUNY Mt Sinai Sch Med, Ruttenberg Canc Ctr, New York, NY 10029 USA. NIH, Immunoregulat Lab, Bethesda, MD 20892 USA. RP Ronai, Z (reprint author), CUNY Mt Sinai Sch Med, Ruttenberg Canc Ctr, 1425 Madison Ave,Room 15-20, New York, NY 10029 USA. RI Ivanov, Vladimir/A-3081-2008; OI Ivanov, Vladimir/0000-0002-2933-6339; RONAI, ZEEV/0000-0002-3859-0400; Kehrl, John/0000-0002-6526-159X FU NCI NIH HHS [CA 51995] NR 61 TC 27 Z9 27 U1 0 U2 1 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD FEB 17 PY 2000 VL 19 IS 7 BP 933 EP 942 DI 10.1038/sj.onc.1203415 PG 10 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 288MW UT WOS:000085567700011 PM 10702802 ER PT J AU Take, H Watanabe, S Takeda, K Yu, ZX Iwata, N Kajigaya, S AF Take, H Watanabe, S Takeda, K Yu, ZX Iwata, N Kajigaya, S TI Cloning and characterization of a novel adaptor protein, CIN85, that interacts with c-Cbl SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE cloning; c-Cbl-interacting protein; adaptor protein; Src homology 3 domain; epidermal growth factor receptor ID EPIDERMAL-GROWTH-FACTOR; T-CELL ACTIVATION; HUMAN HEMATOPOIETIC-CELLS; PREDICTING COILED COILS; TYROSINE PHOSPHORYLATION; PHOSPHATIDYLINOSITOL 3-KINASE; EGF RECEPTOR; IN-VIVO; PROTOONCOGENE PRODUCT; ANTIGEN RECEPTOR AB The c-Cbl protooncogene product is a prominent substrate of protein tyrosine kinases and is rapidly tyrosine-phosphorylated upon stimulation of a wide variety of cell-surface receptors, We have identified a novel c-Cbl-interacting protein termed CIN85 with a molecular mass of 85 kDa which shows similarity to adaptor proteins, CMS and CD2AP, CIN85 mRNA is expressed ubiquitously in normal human tissues and cancer cell lines analyzed. CIN85 was basally associated with c-Cbl. For interaction of CIN85 with c-Cbl, the second SH3 domain of CIN85 was shown to serve as a central player. The CIN85-c-Cbl association was enhanced shortly after stimulation of 293 cells with epidermal growth factor (EGF) and gradually diminished to a basal level, which correlated with a tyrosine phosphorylation level of c-Cbl. Our results suggest that CIN85 may play a specific role in the EGF receptor-mediated signaling cascade via its interaction with c-Cbl. (C) 2000 Academic Press. C1 NHLBI, Hematol Branch, NIH, Bethesda, MD 20892 USA. NHLBI, Pathol Sect, NIH, Bethesda, MD 20892 USA. RP Kajigaya, S (reprint author), NHLBI, Hematol Branch, NIH, Bldg 10,Rm 7C207,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 57 TC 119 Z9 123 U1 0 U2 1 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD FEB 16 PY 2000 VL 268 IS 2 BP 321 EP 328 DI 10.1006/bbrc.2000.2147 PG 8 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 286DB UT WOS:000085427400015 PM 10679202 ER PT J AU Hauger, RL Smith, RD Braun, S Dautzenberg, FM Catt, KJ AF Hauger, RL Smith, RD Braun, S Dautzenberg, FM Catt, KJ TI Rapid agonist-induced phosphorylation of the human CRF receptor, type 1: A potential mechanism for homologous desensitization SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article ID PROTEIN-COUPLED RECEPTORS; XENOPUS-LAEVIS; KINASE; IDENTIFICATION; EXPRESSION; CLONING; CELLS AB Agonist-induced phosphorylation of the human corticotropin-releasing factor type 1 receptor (hCRF(1)-R) was investigated using an influenza hemagglutinin (HA) epitope-tagged receptor transiently expressed in COS-7 cells. The HA-hCRF(1)-R migrated as a broad band (M-r 60,000-70,000) in SDS-PAGE and showed increased mobility (M-r similar to 48,000) after enzymatic deglycosylation with peptide-N-glycosidase F, consistent with the predicted size (47 kDa) of the nonglycosylated HA-hCRF(1)-R protein. A marked increase in HA-hCRF(1)-R phosphorylation was observed in HA-hCRF,R-expressing COS-7 cells exposed to 1 mu M ovine CRF for 5 min, whereas activation of protein kinase A (PRA) by 50 mu M forskolin, or of Ca2+/calmodulin (CaM)dependent kinases by 10 mu M ionomycin, had little effect. These findings are consistent with preliminary data suggesting that CRF1-R phosphorylation mediated by G protein receptor kinase 3 (GRK3), but not by PKA or CaM-dependent kinases, has an important role in the homologous desensitization of brain CRF1-Rs, (C) 2000 Academic Press. C1 Univ Calif San Diego, VA Med Ctr, La Jolla, CA 92093 USA. Univ Calif San Diego, Dept Psychiat, La Jolla, CA 92093 USA. NICHHD, NIH, Endocrinol & Reprod Res Branch, Bethesda, MD 20892 USA. F Hoffmann LaRoche Ltd, Div Pharma, CH-4070 Basel, Switzerland. RP Hauger, RL (reprint author), Univ Calif San Diego, VA Med Ctr, 9500 Gilman Dr, La Jolla, CA 92093 USA. FU NIMH NIH HHS [MH20914-14] NR 29 TC 34 Z9 35 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD FEB 16 PY 2000 VL 268 IS 2 BP 572 EP 576 DI 10.1006/bbrc.2000.2183 PG 5 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 286DB UT WOS:000085427400058 PM 10679245 ER PT J AU Fisher, ER Fisher, B AF Fisher, ER Fisher, B TI Relation of a recurrent intraductal carcinoma (ductal carcinoma in situ) to the primary tumor SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Editorial Material ID SURGICAL ADJUVANT BREAST; IN-SITU; RADIATION-THERAPY; LUMPECTOMY; CANCER; B-17; DCIS C1 NSABP Sci Directors Off, Pittsburgh, PA 15212 USA. Natl Surg Adjuvant Breast & Bowel Project, Pathol Ctr, Pittsburgh, PA USA. RP Fisher, B (reprint author), NSABP Sci Directors Off, 4 Allegheny Ctr,Suite 602, Pittsburgh, PA 15212 USA. NR 19 TC 4 Z9 4 U1 0 U2 0 PU NATL CANCER INSTITUTE PI BETHESDA PA 9030 OLD GEORGETOWN RD, BETHESDA, MD 20814 USA SN 0027-8874 J9 J NATL CANCER I JI J. Natl. Cancer Inst. PD FEB 16 PY 2000 VL 92 IS 4 BP 288 EP 289 DI 10.1093/jnci/92.4.288 PG 2 WC Oncology SC Oncology GA 283UR UT WOS:000085294900001 PM 10675367 ER PT J AU Khalili, H Zhang, FJ Harvey, RG Dipple, A AF Khalili, H Zhang, FJ Harvey, RG Dipple, A TI Mutagenicity of benzo[a]pyrene-deoxyadenosine adducts in a sequence context derived from the p53 gene SO MUTATION RESEARCH-GENETIC TOXICOLOGY AND ENVIRONMENTAL MUTAGENESIS LA English DT Article DE p53 gene; benzo[a]pyrene diol epoxide; deoxyadenosine adducts; lung cancer; mutations ID (+)-ANTI-BENZOPYRENE DIOL EPOXIDE; OLIGONUCLEOTIDE ADDUCTS; DNA-ADDUCTS; IN-VITRO; BENZOPYRENE; MUTAGENESIS; MUTATIONS; METABOLITES; NUCLEOSIDE; CELLS AB Mutations in the human p53 tumor suppressor gene are prominently linked to sporadic cancers in breast, lung and other tissues. Recent research has shown that tobacco-associated cancer in the human lung is related to mutation of the p53 gene mediated by the carcinogen benzo[a]pyrene (BaP), and the mutations are targeted to DNA "hot spots" at specific codons. In order to gain insight into the relation between the structures of the adducts formed by BaP at these sites and their mutagenic activities, we have synthesized site-specifically modified oligo-nucleotide adducts of the active BaP diol epoxide metabolite (anti-BaPDE). This manuscript reports on the mutagenic consequences of replication past anti-BaPDE-deoxy-adenosine adducts located within a sequence context related to codon 157 in exon 5 of the p53 gene. In this sequence context, the adduct derived from the carcinogenic 7R,8S-dihydrodiol 9S,10R-epoxide was much more active as a mutagen than the adduct derived from the noncarcinogenic 7S,8R-dihydrodiol 9R,10S-epoxide and the mutation found most frequently was an A --> G transition. Since previous studies in other sequence contexts have yielded somewhat different findings, these studies further emphasize the key role played by sequence context in determining the mutational properties of carcinogen-DNA adducts. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Univ Chicago, Ben May Inst Canc Res, Chicago, IL 60637 USA. NCI, Frederick Canc Res & Dev Ctr, ABL Basic Res Program, Chem Carcinogenesis Lab, Frederick, MD 21702 USA. RP Harvey, RG (reprint author), Univ Chicago, Ben May Inst Canc Res, MC 6027,5841 S Maryland Ave, Chicago, IL 60637 USA. NR 22 TC 14 Z9 14 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 1383-5718 J9 MUTAT RES-GEN TOX EN JI Mutat. Res. Genet. Toxicol. Environ. Mutagen. PD FEB 16 PY 2000 VL 465 IS 1-2 BP 39 EP 44 DI 10.1016/S1383-5718(99)00203-X PG 6 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology SC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology GA 291XT UT WOS:000085765600004 PM 10708967 ER PT J AU Bianchi, GD Cerhan, JR Parker, AS Putnam, SD See, WA Lynch, CF Cantor, KP AF Bianchi, GD Cerhan, JR Parker, AS Putnam, SD See, WA Lynch, CF Cantor, KP TI Tea consumption and risk of bladder and kidney cancers in a population-based case-control study SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Article DE bladder neoplasms; case-control studies; diet; kidney neoplasms; tea ID DRINKING-WATER SOURCE; RENAL-CELL CARCINOMA; LOWER URINARY-TRACT; DIETARY FACTORS; FLUID INTAKE; BLACK TEA; LUNG TUMORIGENESIS; LIFE-STYLE; A/J MICE; COFFEE AB Recent epidemiologic studies have suggested that tea may be protective against cancers of the urinary tract. The authors examined the association between usual adult tea consumption and risk of bladder and kidney cancers in a population-based case-control study that included 1,452 bladder cancer cases, 406 kidney cancer cases, and 2,434 controls. For bladder cancer, the age- and sex-adjusted odds ratios (OR) (95% confidence intervals (CI)) referent to nonusers of tea were 0.9 (0.7, 1.1) for <1.0 cup/day, 1.0 (0.8, 1.2) for 1.0-2.6 cups/day, and 0.9 (0.7, 1.1) for >2.6 cups/day (cutpoints for users based on the tertile distribution among controls). When more extreme cutpoints were used, persons who consumed >5 cups/day (>90th percentile) had a suggestive decreased risk (OR = 0.7; 95% CI 0.5, 1.0), but there was no evidence of a dose-response relation. in analyses stratified by median total beverage intake (2.6 liters/day), there was an inverse association with tea use among persons who consumed less than the median (OR = 0.5; 95% CI 0.3, 0.8) but no association for persons who consumed at or above the median. In contrast, for kidney cancer, there was no association with tea use. Adjustment for site-specific risk factors did not alter these results. This study offers only minimal support for an inverse association between tea consumption and bladder or kidney cancer risk. C1 Mayo Clin & Mayo Fdn, Dept Hlth Sci Res, Rochester, MN 55905 USA. Univ Iowa, Coll Med, Dept Prevent Med & Environm Hlth, Iowa City, IA 52242 USA. Univ Iowa Hosp & Clin, Dept Urol, Iowa City, IA 52242 USA. Mayo Clin, Dept Hlth Sci Res, Rochester, MN USA. NCI, Occupat Epidemiol Branch, Bethesda, MD 20892 USA. RP Cerhan, JR (reprint author), Mayo Clin & Mayo Fdn, Dept Hlth Sci Res, 200 1st St SW, Rochester, MN 55905 USA. FU NCI NIH HHS [K07 CA64220, NCI-NO1-CP-51026, NCI-NO1-CP-85614] NR 41 TC 48 Z9 49 U1 1 U2 2 PU JOHNS HOPKINS UNIV SCHOOL HYGIENE PUB HEALTH PI BALTIMORE PA 111 MARKET PLACE, STE 840, BALTIMORE, MD 21202-6709 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD FEB 15 PY 2000 VL 151 IS 4 BP 377 EP 383 PG 7 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 286XU UT WOS:000085474800006 PM 10695596 ER PT J AU Boutron-Ruault, HC Rabkin, CS AF Boutron-Ruault, HC Rabkin, CS TI Re: "Controls who experienced hypothetical causal intermediates should not be excluded from case-control studies" SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Letter ID COLORECTAL-CANCER; CIGARETTE-SMOKING; RISK; ADENOMA; MEN; ALCOHOL C1 Conservatoire Natl Arts & Metiers, Inst Sci & Tech Nutr & Alimentat, F-75003 Paris, France. NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. RP Boutron-Ruault, HC (reprint author), Conservatoire Natl Arts & Metiers, Inst Sci & Tech Nutr & Alimentat, 2 Rue Conte, F-75003 Paris, France. NR 9 TC 1 Z9 1 U1 1 U2 1 PU JOHNS HOPKINS UNIV SCHOOL HYGIENE PUB HEALTH PI BALTIMORE PA 111 MARKET PLACE, STE 840, BALTIMORE, MD 21202-6709 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD FEB 15 PY 2000 VL 151 IS 4 BP 436 EP 436 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 286XU UT WOS:000085474800013 ER PT J AU Cohen, HJ Pieper, CF Hanlon, JT Wall, WE Burchett, BM Havlik, RJ AF Cohen, HJ Pieper, CF Hanlon, JT Wall, WE Burchett, BM Havlik, RJ TI Calcium channel blockers and cancer SO AMERICAN JOURNAL OF MEDICINE LA English DT Article ID DRUG-USE; RISK; ANTAGONISTS; INCREASE; MORTALITY; HEALTH; BLACK; WOMEN AB PURPOSE: We sought to explore the relation that has been previously reported between calcium channel blockers and an increased risk of cancer. SUBJECTS AND METHODS: We followed 3,511 participants, age 65 years or older, in the Duke Established Populations for Epidemiologic Studies of the Elderly for up to 10 years. Information about use of medications was obtained at baseline and 3 and 6 years later. Information about hospitalization for cancer, or death from cancer, was obtained from Health Care Financing Administration data and death certificates. RESULTS: Of the 133 users of calcium channel blockers, 16 (12%) developed cancer, compared with 548 (16%) of 3,378 nonusers (hazard ratio = 0.9; 95% confidence interval, 0.5 to 1.5). Adjusting for baseline and time-dependent covariates, such as race, diabetes, or blood pressure, for dose or class of calcium channel blockers, or for length of follow-up, had no effect. CONCLUSIONS: Use of calcium channel blockers does not appear to be related to cancer risk. Earlier reports showing such a relation may have been the result of chance. (C) 2000 by Excerpta Medica, Inc. C1 Duke Univ, Med Ctr, Ctr Study Aging & Human Dev, Durham, NC 27710 USA. Duke Univ, Med Ctr, Dept Med, Div Geriatr, Durham, NC 27710 USA. Duke Univ, Med Ctr, Dept Med, Div Gen Internal Med, Durham, NC 27710 USA. Duke Univ, Med Ctr, Dept Family Med, Div Biometry, Durham, NC 27710 USA. Vet Affairs Med Ctr, Ctr Geriatr Res Educ & Clin, Durham, NC 27705 USA. Univ Minnesota, Coll Pharm, Minneapolis, MN 55455 USA. NIA, Epidemiol Demog & Biometry Program, Bethesda, MD 20892 USA. RP Cohen, HJ (reprint author), Duke Univ, Med Ctr, Ctr Study Aging & Human Dev, Box 3003, Durham, NC 27710 USA. FU NIA NIH HHS [N01-AG-1-2102] NR 26 TC 13 Z9 13 U1 0 U2 0 PU EXCERPTA MEDICA INC PI NEW YORK PA 245 WEST 17TH STREET, NEW YORK, NY 10011 USA SN 0002-9343 J9 AM J MED JI Am. J. Med. PD FEB 15 PY 2000 VL 108 IS 3 BP 210 EP 215 DI 10.1016/S0002-9343(99)00447-7 PG 6 WC Medicine, General & Internal SC General & Internal Medicine GA 284LU UT WOS:000085334500005 PM 10723975 ER PT J AU Horne, MK Mayo, DJ Cannon, RO Chen, CC Shawker, TH Chang, R AF Horne, MK Mayo, DJ Cannon, RO Chen, CC Shawker, TH Chang, R TI Intraclot recombinant tissue plasminogen activator in the treatment of deep venous thrombosis of the lower and upper extremities SO AMERICAN JOURNAL OF MEDICINE LA English DT Article ID RANDOMIZED TRIAL; VEIN THROMBOSIS; STREPTOKINASE; HEPARIN; REFLUX C1 NHLBI, Warren G Magnuson Clin Ctr, Dept Clin Pathol, NIH, Bethesda, MD 20892 USA. NHLBI, Warren G Magnuson Clin Ctr, Dept Nucl Med, NIH, Bethesda, MD 20892 USA. NHLBI, Warren G Magnuson Clin Ctr, Dept Diagnost Radiol, NIH, Bethesda, MD 20892 USA. NHLBI, Cardiol Branch, NIH, Bethesda, MD 20892 USA. RP Horne, MK (reprint author), NHLBI, Warren G Magnuson Clin Ctr, Dept Clin Pathol, NIH, Room 2C306,Bldg 10,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 10 TC 27 Z9 28 U1 0 U2 0 PU EXCERPTA MEDICA INC PI NEW YORK PA 245 WEST 17TH STREET, NEW YORK, NY 10011 USA SN 0002-9343 J9 AM J MED JI Am. J. Med. PD FEB 15 PY 2000 VL 108 IS 3 BP 251 EP 255 DI 10.1016/S0002-9343(99)00407-6 PG 5 WC Medicine, General & Internal SC General & Internal Medicine GA 284LU UT WOS:000085334500011 PM 10723981 ER PT J AU Cohen, OJ AF Cohen, OJ TI Antiretroviral therapy: Time to think strategically SO ANNALS OF INTERNAL MEDICINE LA English DT Editorial Material ID HUMAN-IMMUNODEFICIENCY-VIRUS; HIV-1 INFECTION; DISEASE; LYMPHOCYTES; RESPONSES; STAGE C1 NIAID, Bethesda, MD 20892 USA. RP Cohen, OJ (reprint author), NIAID, 31 Ctr Dr MSC 2520,Bldg 31,Room 7A05, Bethesda, MD 20892 USA. NR 20 TC 11 Z9 11 U1 0 U2 0 PU AMER COLL PHYSICIANS PI PHILADELPHIA PA INDEPENDENCE MALL WEST 6TH AND RACE ST, PHILADELPHIA, PA 19106-1572 USA SN 0003-4819 J9 ANN INTERN MED JI Ann. Intern. Med. PD FEB 15 PY 2000 VL 132 IS 4 BP 320 EP 322 PG 3 WC Medicine, General & Internal SC General & Internal Medicine GA 285CY UT WOS:000085372900009 PM 10681288 ER PT J AU Adachi, K Katsuyama, M Song, S Oka, T AF Adachi, K Katsuyama, M Song, S Oka, T TI Genomic organization, chromosomal mapping and promoter analysis of the mouse selenocysteine tRNA gene transcription-activating factor (mStaf) gene SO BIOCHEMICAL JOURNAL LA English DT Article DE gene structure; promoter activity; Sp1; transcription start site ID TRANSFER-RNA GENE; INSITU HYBRIDIZATION; MAMMARY-GLAND; SP1; EXPRESSION; FEATURES; ELEMENTS; CLONING; ZNF76; STAF AB mStaf is a zinc-finger protein that activates the transcription of the mouse selenocysteine tRNA gene. The mStaf gene is approx. 35 kb long and split into 16 exons, All exon-intron junction sequences conform to the GT/AG rule. The transcription start site is located 83 bp upstream of the initiation codon. Chromosomal mapping localized the gene to mouse chromosome 7, region E3-F1, Sequence analysis of the proximal promoter region revealed several potential regulatory elements; these include the recognition elements of Sp1, Nkx, CP2, E2A, SIF (SIS-inducible factor), TFII-I and cAMP-responsive element (CRE), but no TATA sequences. Transfection experiments demonstrated that the 5'-flanking region (-1894 to + 37) of the mStaf gene drives transcription in mouse NMuMG cells and that a construct containing a fragment from -387 to + 37 showed the highest transcriptional activity. Deletion and mutation experiments suggested that four Spl sites played an important role for the basal promoter activity. Furthermore, electrophoretic mobility-shift assays demonstrated that Sp3 but not other Sp (specificity protein) family members binds to three of the Spl sites. Our present study suggests that Sp3 is involved in the basal transcriptional activation of the mStaf gene. C1 NIDDKD, Lab Genet & Physiol, NIH, Bethesda, MD 20892 USA. RP Oka, T (reprint author), NIDDKD, Lab Genet & Physiol, NIH, Room 106,Bldg 8, Bethesda, MD 20892 USA. NR 36 TC 2 Z9 3 U1 0 U2 0 PU PORTLAND PRESS PI LONDON PA 59 PORTLAND PLACE, LONDON W1N 3AJ, ENGLAND SN 0264-6021 J9 BIOCHEM J JI Biochem. J. PD FEB 15 PY 2000 VL 346 BP 45 EP 51 DI 10.1042/0264-6021:3460045 PN 1 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 289TY UT WOS:000085639100007 PM 10657238 ER PT J AU Sugiyama, K Chen, Z Lee, YS Kador, PF AF Sugiyama, K Chen, Z Lee, YS Kador, PF TI Isolation of a non-covalent aldose reductase-nucleotide-inhibitor complex SO BIOCHEMICAL PHARMACOLOGY LA English DT Article DE aldose reductase; diabetes; aldose reductase inhibitors; mechanism of action; non-covalent binding ID ELECTROSPRAY MASS-SPECTROMETRY; DIABETIC COMPLICATIONS; ACTIVE-SITE; BINDING; NONCOVALENT; SORBINIL; KINETICS AB A method fur the isolation of an intact, non-covalent complex formed by the interaction of aldose reductase, NADP(H) nucleotide, and inhibitor has been developed to aid in the discovery and development of novel aldose reductase inhibitors. In the complexes isolated, both the carboxylic acid-containing inhibitor tolrestat and the spirohydantoin-containing inhibitor AL1576 (2,7-difluorospirofluorene-9,5'-imidazolidine-2',4'-dione) tightly bound in a 1:1 ratio to aldose reductase complexed with either NADPH or NAD(+). Inhibitor binding to either the enzyme-NADP(+) or enzyme-NADPH complex appeared to be equal and pH-dependent, with maximum binding observed at a pH range of 7 to 8.5 where the inhibitors are ionized. These results indicated that the charge state of the cofactor (NADPH vs NADP(+)) is not critical for inhibitor binding to aldose reductase. Molecular modeling studies suggested that His110 plays a crucial rule in directing charged inhibitors containing either a carboxylate or an ionizable hydantoin group to the active site of aldose reductase by providing charge interaction. BIOCHEM PHARMACOL 59;4:329-336, 2000. (C) 2000 Elsevier Science Inc. C1 NEI, Lab Ocular Therapeut, NIH, Bethesda, MD 20892 USA. RP Kador, PF (reprint author), NEI, Lab Ocular Therapeut, NIH, Bldg 10,Room 10B-11,10 Ctr Dr,MSC 1850, Bethesda, MD 20892 USA. NR 28 TC 10 Z9 11 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0006-2952 J9 BIOCHEM PHARMACOL JI Biochem. Pharmacol. PD FEB 15 PY 2000 VL 59 IS 4 BP 329 EP 336 DI 10.1016/S0006-2952(99)00332-9 PG 8 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 273PA UT WOS:000084715000003 PM 10644040 ER PT J AU Szczepanik, MB Pande, P Laux, WG Debrabant, A Dwyer, D Johnson, RA AF Szczepanik, MB Pande, P Laux, WG Debrabant, A Dwyer, D Johnson, RA TI Inhibition of Leishmania donovani 3'-nucleotidase by 9-substituted adenine derivatives. SO BIOCHEMISTRY LA English DT Meeting Abstract C1 SUNY Stony Brook, Hlth Sci Ctr, Stony Brook, NY 11794 USA. NIAID, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD FEB 15 PY 2000 VL 39 IS 6 MA 36 BP 1549 EP 1550 PG 2 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 285NX UT WOS:000085396100079 ER PT J AU Du, X Welch, AR Coleman, WG AF Du, X Welch, AR Coleman, WG TI Purification and characterization of the ADP-D-glycero-D-mannoheptose synthetase from Helicobactor pylori. SO BIOCHEMISTRY LA English DT Meeting Abstract C1 NIDDK, Lab Biochem & Genet, NIH, Bethesda, MD 20892 USA. Diagnon Corp, Dept Res & Dev, Rockville, MD USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD FEB 15 PY 2000 VL 39 IS 6 MA 168 BP 1579 EP 1580 PG 2 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 285NX UT WOS:000085396100209 ER PT J AU Wang, LH Yang, XY Kirken, RA Resau, JH Farrar, WL AF Wang, LH Yang, XY Kirken, RA Resau, JH Farrar, WL TI Targeted disruption of Stat6 DNA binding activity by an oligonucleotide decoy blocks IL-4-driven T(H)2 cell response SO BLOOD LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; TRANSCRIPTION FACTOR DECOY; CYTOKINE GENE-EXPRESSION; NF-KAPPA-B; T-CELLS; IN-VIVO; BLOOD-PRESSURE; CIS-ELEMENTS; IL-4; TH2 AB The transcription factor, signal transducer and activator of transcription (Stat) 6, regulates T(H)2-lymphocyte activity by controlling the expression and responsiveness to interleukin (IL)-4, which plays a key role in numerous allergic maladies. Therefore, we sought to use a phosphorothiolate cis-element decoy to target disruption of Stat6 transcriptional activity. Here we showed that the State decoy potently ablated the messenger RNA expression and production of IL-4, but not of several other cytokines. The Stat6 decoy functionally disrupted IL-4-inducible cell proliferation of murine T(H)2 cells and primary human CD4(+) T lymphocytes, Specificity of the decoy was demonstrated by its ability to directly block State binding to a cis-element probe and transactivation, but not affect Stat6 tyrosine phosphorylation or expression of the IL-4 receptor chains, Moreover the decoy failed to inhibit non-Stat6-dependent signaling pathways since IL-2 was competent to induce cell proliferation and activation of Stats 1, 3, and 5a/b, With the use of laser scanning confocal microscopy, fluorescently tagged Stats decoy was detectable in the cytoplasm and nucleus; however, greater levels of oligonucleotide were present in the latter following IL-4 treatment. Taken together, these data suggest that IL-4-driven T(H)2 cell activity can be preferentially restricted via targeted disruption of Stat6 by a novel and specific decoy strategy that may possess gene therapeutic potential, (Blood, 2000;95: 1249-1257) (C) 2000 by The American Society of Hematology. C1 NCI, Frederick Canc Res & Dev Ctr, Cytokine Mol Mechanisms Sect, Mol Immunoregulat Lab,Div Basic Sci, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, Intramural Res Support Program, Frederick, MD 21702 USA. Univ Texas, Dept Integrat Biol Pharmacol & Physiol, Houston, TX USA. NCI, ABL Basic Res Program, Frederick, MD 21701 USA. RP Farrar, WL (reprint author), NCI, Frederick Canc Res & Dev Ctr, Cytokine Mol Mechanisms Sect, Mol Immunoregulat Lab,Div Basic Sci, POB B,Bldg 560,Room 31-68, Frederick, MD 21702 USA. FU NCI NIH HHS [N01CO56000] NR 59 TC 55 Z9 57 U1 0 U2 2 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1200 19TH ST, NW, STE 300, WASHINGTON, DC 20036-2422 USA SN 0006-4971 J9 BLOOD JI Blood PD FEB 15 PY 2000 VL 95 IS 4 BP 1249 EP 1257 PG 9 WC Hematology SC Hematology GA 283DV UT WOS:000085261600022 PM 10666197 ER PT J AU Fernandez, E AF Fernandez, E TI Thymocyte development past the CD4(+)CD8(+) stage requires an active p38 mitogen-activated protein kinase SO BLOOD LA English DT Article ID T-CELL DEVELOPMENT; MAP KINASE; LYMPHOCYTE; STRESS; DIFFERENTIATION; PHOSPHORYLATION; EXPRESSION; INHIBITION; SELECTION; INTERLEUKIN-2 AB Activation of the p38 mitogen-activated protein kinase (MAPK) pathway is important for some T-cell functions, but its role in intrathymic development is unclear. To investigate the function of p38 MAPK during the late stages of thymocyte differentiation, pharmacologic and genetic manipulations were used to inhibit p38 MAPK activity in developing thymocytes. Ligation of the T-cell antigen receptor (TCR) on either thymocytes or a thymocyte cell line resulted in p38 MAPK activation. Selective pharmacologic inhibition of p38 MAPK activity with the pyridinyl imidazole drug SB203580 severely impaired the development of mature CD4(+) and CD8(+) single positive (SP) thymocytes from their CD4(+)CD8(+) double positive (DP) precursors in fetal thymic organ culture (FTOC). Further, pharmacologic or genetic suppression of p38 MAPK activity, the latter achieved by overexpressing a catalytically inactive p38 MAPK, resulted in a blockade of the DP-to-SP transition of a thymocyte cell line in a novel in vitro differentiation assay. Taken together, these data constitute the first demonstration that p38 MAPK plays a critical role in the DP-to-SP differentiation of thymocytes during late intrathymic development. C1 NIAID, Struct Biol Sect, NIH, Rockville, MD USA. RP Fernandez, E (reprint author), Univ Autonoma Madrid, CSIC, Ctr Biol Mol Severo Ochoa, E-28049 Madrid, Spain. NR 40 TC 11 Z9 11 U1 0 U2 0 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1200 19TH ST, NW, STE 300, WASHINGTON, DC 20036-2422 USA SN 0006-4971 J9 BLOOD JI Blood PD FEB 15 PY 2000 VL 95 IS 4 BP 1356 EP 1361 PG 6 WC Hematology SC Hematology GA 283DV UT WOS:000085261600036 PM 10666211 ER PT J AU Kagami, S Nakajima, H Kumano, K Suzuki, K Suto, A Imada, K Davey, HW Saito, Y Takatsu, K Leonard, WJ Iwamoto, I AF Kagami, S Nakajima, H Kumano, K Suzuki, K Suto, A Imada, K Davey, HW Saito, Y Takatsu, K Leonard, WJ Iwamoto, I TI Both Stat5a and Stat5b are required for antigen-induced eosinophil and T-cell recruitment into the tissue SO BLOOD LA English DT Article ID INDUCED AIRWAY HYPERRESPONSIVENESS; COLONY-STIMULATING FACTOR; MAMMARY-GLAND FACTOR; GENE-EXPRESSION; SIGNAL-TRANSDUCTION; CYTOKINE RECEPTORS; ACTIVATES STAT5; NATURAL-KILLER; DEFICIENT MICE; MOUSE AIRWAYS AB Antigen-induced eosinophil recruitment into the airways of sensitized mice is mediated by CD4(+) T cells and their cytokines, especially IL-5. In this study, we found that the antigen-induced airway eosinophilia was diminished in Stat5a-deficient (Stat5a(-/-)) mice and Stat5b-deficient (Stat5b(-/-)) mice. We also found that antigen-induced CD4(+) T-cell infiltration and IL-5 production in the airways were diminished in Stat5a(-/-) mice and Stat5b(-/-) mice. Moreover, antigen-induced proliferation of splenocytes was diminished in Stat5a(-/-) mice and Stat5b(-/-) mice, suggesting that the generation of antigen-primed T cells may be compromised in Stat5a(-/-) mice and Stat5b(-/-) mice and this defect may account for the diminished antigen induced T-cell infiltration into the airways. Interestingly, IL-4 and IL-5 production from anti-CD3-stimulated splenocytes was diminished in Stat5a(-/-) mice and Stat5b(-/-) mice. However, antigen-specific IgE and IgG1 production was diminished in Stat5a(-/-) mice but not in Stat5b(-/-) mice, whereas antigen-specific IgG2a production was increased in Stat5a(-/-) mice, suggesting the enhanced Th1 responses in Stat5a(-/-) mice. Finally, we found that eosinophilo-poiesis induced by the administration of recombinant IL-5 was also diminished in Stat5a(-/-) mice and Stat5b(-/-) mice. Together, these results indicate that both Stat5a and Stat5b are essential for induction of antigen-induced eosinophil recruitment into the airways and that the defects in antigen-induced eosinophil recruitment in Stat5a(-/-) mice and Stat5b(-/-) mice result from both impaired IL-5 production in the airways and diminished IL-5 responsiveness of eosinophils. (C) 2000 by The American Society of Hematology. C1 Chiba Univ, Sch Med, Dept Internal Med 2, Chiba 2608670, Japan. NHLBI, Lab Mol Immunol, Bethesda, MD 20892 USA. AgResearch, Hamilton, New Zealand. Univ Tokyo, Inst Med Sci, Dept Immunol, Tokyo, Japan. RP Nakajima, H (reprint author), Chiba Univ, Sch Med, Dept Internal Med 2, 1-8-1 Inohana, Chiba 2608670, Japan. NR 60 TC 60 Z9 60 U1 0 U2 0 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1200 19TH ST, NW, STE 300, WASHINGTON, DC 20036-2422 USA SN 0006-4971 J9 BLOOD JI Blood PD FEB 15 PY 2000 VL 95 IS 4 BP 1370 EP 1377 PG 8 WC Hematology SC Hematology GA 283DV UT WOS:000085261600038 PM 10666213 ER PT J AU Sakai, A Marti, GE Caporaso, N Pittaluga, S Touchman, JW Fend, F Raffeld, M AF Sakai, A Marti, GE Caporaso, N Pittaluga, S Touchman, JW Fend, F Raffeld, M TI Analysis of expressed immunoglobulin heavy chain genes in familial B-CLL SO BLOOD LA English DT Article ID CHRONIC LYMPHOCYTIC-LEUKEMIA; CHRONIC LYMPHATIC-LEUKEMIA; V-H GENES; SOMATIC HYPERMUTATION; CHROMOSOMAL-ABNORMALITIES; INSITU HYBRIDIZATION; CELLS; REPERTOIRE; SELECTION; DELETIONS AB In this study, we wished to determine whether familial chronic lymphocytic leukemia of B-cell phenotype (CLL) shares with sporadic B-CLL the same immunoglobulin (Ig) heavy chain variable region (VH) gene usage and occurrence of somatic mutation, to gain insight into the pathogenetic relatedness of these epidemiologically distinct forms of CLL. We therefore analyzed the expressed Ig heavy chain genes in 23 cases (11 families) of familial CLL, and compared these results with data previously reported for sporadic CLL, In addition, we assessed the relationship of the occurrence of somatic mutation to several clinical and phenotypic features. The distribution of V genes among these cases was similar to that observed in sporadic CLL: VH3 > VH1 > VH4. Thirteen of the 23 cases (57%) showed germ line VH gene sequences, whereas somatic mutations were detected in 10 cases (43%), The average mutation frequency of these latter 10 cases of was 6.7% (ranging from 1.7% to 8.8%), and evidence of antigen selection was noted in 6. Intraclonal variation, followed by clonal evolution and the appearance of a second clone over a 20 year period was observed in 1 case, suggesting that mutations can continue to accumulate after neoplastic transformation. The presence of somatic mutations correlated with age at presentation, low white blood cell (WBC) count, and low fluorescence intensity of surface CD5, and the potential significance of these relationships is discussed. Our data indicate that familial end sporadic B-CLL display a similar pattern of immunoglobulin gene usage and frequency of somatic mutation, and are consistent with a common ontogeny and immunogenetic origin for these 2 epidemiologically distinct forms of CLL. (C) 2000 by The American Society of Hematology. C1 NCI, Hematopathol Sect, Pathol Lab, NIH, Bethesda, MD 20892 USA. US FDA, Ctr Biol Evaluat & Res, Flow & Image Cytometry Sect, Lab Med & Mol Genet,Div Cell & Gene Therapies, Bethesda, MD USA. NCI, Genet Epidemiol Branch, NIH, Rockville, MD USA. NIH, Intramural Sequencing Ctr, Gaithersburg, MD USA. Natl Human Genome Res Inst, NIH, Bethesda, MD USA. RP Raffeld, M (reprint author), NCI, Hematopathol Sect, Pathol Lab, NIH, Bldg 10,Room 2N110,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 42 TC 49 Z9 51 U1 0 U2 2 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1200 19TH ST, NW, STE 300, WASHINGTON, DC 20036-2422 USA SN 0006-4971 J9 BLOOD JI Blood PD FEB 15 PY 2000 VL 95 IS 4 BP 1413 EP 1419 PG 7 WC Hematology SC Hematology GA 283DV UT WOS:000085261600044 PM 10666219 ER PT J AU Heiskanen, MA Bittner, ML Chen, YD Khan, J Adler, KE Trent, JM Meltzer, PS AF Heiskanen, MA Bittner, ML Chen, YD Khan, J Adler, KE Trent, JM Meltzer, PS TI Detection of gene amplification by genomic hybridization to cDNA microarrays SO CANCER RESEARCH LA English DT Article ID C-MYC; CELLS; DEPOSITION AB Gene amplification is one of the major mechanisms of oncogene activation in tumorigenesis, To facilitate the identification of genes mapping to amplified regions, we have used a technique based on the hybridization of total genomic DNA to cDNA microarrays, To aid detection of the weak signals generated in this complex hybridization, we have used a tyramide-based technique that allows amplification of a fluorescent signal up to 1000-fold. Dilution experiment suggests that amplifications of 5-fold and higher can be detected by this approach. The technique was validated using cancer cell lines with several known gene amplifications, such as those affecting MYC, MYCN, ERBB2, and CDK4, In addition to the detection of the known amplifications, we identified a novel amplified gene, ZNF133, in the neuroblastoma cell line NGP, Hybridization of NGP cDNA on an identical array also revealed over expression of ZNF133. parallel analysis of genomic DNA for copy number and cDNA for expression now provides rapid approach to the identification of amplified genes and chromosomal regions in tumor cells. C1 Natl Human Genome Res Inst, Canc Gent Branch, NIH, Bethesda, MD 20892 USA. DuPont NEN Life Sci Prod Inc, Boston, MA 02118 USA. RP Meltzer, PS (reprint author), Natl Human Genome Res Inst, Canc Gent Branch, NIH, 49 Convent Dr,MSC 4470, Bethesda, MD 20892 USA. RI Khan, Javed/P-9157-2014 OI Khan, Javed/0000-0002-5858-0488 NR 18 TC 77 Z9 82 U1 1 U2 1 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD FEB 15 PY 2000 VL 60 IS 4 BP 799 EP 802 PG 4 WC Oncology SC Oncology GA 287JQ UT WOS:000085503100006 PM 10706083 ER PT J AU Zhang, ZQ Liu, Q Lantry, LE Wang, Y Kelloff, GJ Anderson, MW Wiseman, RW Lubet, RA You, M AF Zhang, ZQ Liu, Q Lantry, LE Wang, Y Kelloff, GJ Anderson, MW Wiseman, RW Lubet, RA You, M TI A germ-line p53 mutation accelerates pulmonary tumorigenesis: p53-independent efficacy of chemopreventive agents green tea or dexamethasone/myo-inositol and chemotherapeutic agents taxol or adriamycin SO CANCER RESEARCH LA English DT Article ID TUMOR-SUPPRESSOR GENE; FRAUMENI-SYNDROME; MOLECULAR CARCINOGENESIS; LUNG TUMORIGENESIS; A/J MICE; CANCER; SUSCEPTIBILITY; CONSTITUENT; PROGRESSION; DEFICIENT AB Recent evidence indicates that individuals with a p53 germ-line mutation (Li-Fraumeni syndrome) have a 50% risk of developing lung cancer by age 60, In this study, p53 heterozygous knockout mice and p53 transgenic mice carrying a dominant negative mutant were crossed with the A/J mouse, which is highly susceptible to lung tumor induction, to investigate whether a p53 germ-line mutation is a predisposing gene for carcinogen-induced pulmonary adenomas in mice. The number of lung tumors was not significantly increased in (TSG-p53 x A/J)F-1 p53 heterozygous knockout mice as compared with that in (TSG-p53 x A/J)F-1 wt mice 16 weeks after exposure to N-nitrosomethylurea (MNU). In contrast, an average of 22 lung tumors were observed in (UL53-3 x A/J)F-1 mice carrying a mutant p53 transgene (135Valp53) compared with an average of 7 lung tumors seen in (UL53-3 x A/J)F-1 wt mice after treatment with N-nitrosomethylurea. Similar enhancement of lung tumor multiplicity (similar to 3-fold) was seen when mutant versus wt mice were treated with the tobacco-related carcinogens benzo[a]pyrene or 4-(methylnitro-samino)-1-(3-pyridyl)-1-butanone. These results suggest that the mutant p53 transgene may have a dominant negative effect on the wt p53. The potential usefulness of this new mouse model in lung cancer chemoprevention and chemotherapy was examined. The chemopreventive efficacy of the green tea or a combination of dietary dexamethasone and myoinositol and the chemotherapeutic efficacy of Taxol or Adriamycin was examined in wt mice or mice with a mutation in the p53 gene. Mice treated with dexamethasone/myo-inositol and green tea displayed an average of 70 and 50% inhibition of lung tumors, respectively, regardless of p53 status, Similarly, when mice bearing established lung adenomas were treated with Taxol or Adriamycin, a decrease in tumor volume of similar to 70% was observed independent of p53 mutation status. Thus, the (UL53-3 x A/J)F-1 p53 transgenic mouse seems to be an excellent model for human carriers of p53 germ-line mutations (Li-Fraumeni syndrome). Furthermore, the lung adenomas generated in this model possess mutations in both the K-ras proto-oncogene and the p53 tumor suppressor gene. This model should prove directly useful for chemoprevention and chemotherapy studies. C1 Med Coll Ohio, Dept Pathol, Toledo, OH 43699 USA. NCI, Chemoprevent Branch, Bethesda, MD 20892 USA. Univ Cincinnati, Dept Environm Hlth, Cincinnati, OH 45267 USA. NIEHS, Mol Carcinogenesis Lab, Res Triangle Pk, NC 27709 USA. RP You, M (reprint author), Med Coll Ohio, Dept Pathol, Hlth Educ Bldg,Room 202,3055 Arlington Ave, Toledo, OH 43614 USA. FU NCI NIH HHS [CA58554, CA78797, CN65111] NR 39 TC 81 Z9 83 U1 1 U2 2 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD FEB 15 PY 2000 VL 60 IS 4 BP 901 EP 907 PG 7 WC Oncology SC Oncology GA 287JQ UT WOS:000085503100026 PM 10706103 ER PT J AU Yuan, BZ Keck-Waggoner, C Zimonjic, DB Thorgeirsson, SS Popescu, NC AF Yuan, BZ Keck-Waggoner, C Zimonjic, DB Thorgeirsson, SS Popescu, NC TI Alterations of the FHIT gene in human hepatocellular carcinoma SO CANCER RESEARCH LA English DT Article ID SQUAMOUS-CELL CARCINOMAS; FRAGILE SITES; FORMER SMOKERS; LUNG-CANCER; FRA3B; BREAKPOINT; DELETION; REGION; 3P14 AB FHIT (fragile histidine triad), a candidate tumor suppressor gene, encompasses FRA3B, a region with the highest fragility in the human genome, and is altered in a large number of human cancers, particularly those of epithelial cell origin and associated with known carcinogenic agents. Human hepatocellular carcinoma (HCC), a major cancer worldwide, is closely related to carcinogenic agents such as hepatitis B and C virus infections, dietary aflatoxin, alcohol consumption, and exposure to chemical carcinogens. To assess the extent and the nature of the FHIT gene alterations and their implications in the development of HCC, several cell lines and primary tumors were cytologically and molecularly examined. The FHIT gene is expressed in normal hepatic cells and is not expressed or is abnormally expressed in cultured tumor cells derived from HCC. Down-regulation of the FHIT gene was detected by Northern blot analysis in 9 of 14 cell lines. However, neither abnormal FHIT transcripts nor point mutations in DNA sequences of reverse transcription-PCR products (exons 2-9) were identified. Expression of FHIT protein was not detected by immunostaining in 5 of 10 primary tumors. Pour cell lines showing mRNA down-regulation did not express FHIT protein as demonstrated by Western blot analysis. Allelic loss of intron 5 of the FHIT gene was detected in 10 of 34 informative samples from primary tumors. Structural alterations of chromosome 3p were identified in 8 of 13 HCC cell lines. Deletions or translocations involving region 3p314.2 were identified by fluorescence in situ hybridization with a YAC850A6 probe spanning the FHIT locus on chromosomes derived from cell lines with an abnormal FHIT gene expression. These combined results indicate that the FHIT gene is a frequent target and may be implicated in a subset of liver cancers. C1 NCI, Expt Carcinogenesis Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Popescu, NC (reprint author), NCI, Expt Carcinogenesis Lab, Div Basic Sci, NIH, Bldg 37,Room 3C05,37 Convent Dr,MSC 4255, Bethesda, MD 20892 USA. NR 28 TC 38 Z9 55 U1 0 U2 2 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD FEB 15 PY 2000 VL 60 IS 4 BP 1049 EP 1053 PG 5 WC Oncology SC Oncology GA 287JQ UT WOS:000085503100046 PM 10706123 ER PT J AU Lehman, TA Haffty, BG Carbone, CJ Bishop, LR Gumbs, AA Krishnan, S Shields, PG Modali, R Turner, BC AF Lehman, TA Haffty, BG Carbone, CJ Bishop, LR Gumbs, AA Krishnan, S Shields, PG Modali, R Turner, BC TI Elevated frequency and functional activity of a specific germ-line p53 intron mutation in familial breast cancer SO CANCER RESEARCH LA English DT Article ID FRAUMENI-SYNDROME; TUMOR-SUPPRESSOR; BRCA1 MUTATIONS; GENE; TP53; APOPTOSIS; GROWTH; CELLS; WOMEN; SUSCEPTIBILITY AB Previous studies have determined that the frequency of germ-line p53 mutations in familial breast cancer patients is 1 % or less, but these reports have not investigated the importance of polymorphic intron base changes in the p53 gene. Therefore, we investigated the frequency of both exon and intron germ-line p53 base changes in 42 breast canter patients with a strong family history of breast cancer. The mean age of presentation of these patients was 44.0 years (range, 29-69), and 12 of 42 (29%) were of known Ashkenazi ancestry, Purified DNA obtained from the 42 index cases was screened for germ-line p53 mutations in exons 2-11 and surrounding introns using a combination of intron based primers for PCR-single strand conformation polymorphism analysis, direct sequencing, and microarray sequencing using the Affymetrix p53 gene chip methodology, Morphological analysis of apoptosis and cell survival determination were performed on EBV-immortalized Lymphoblastoid cell lines from patients with the p53 intron 6 mutation. A germ-line mutation in the p53 gene at nucleotide 13964 with a G to C base change (13964(GC)) was identified in 3 of 42 (7.1%) hereditary breast cancer patients, Two patients were heterozygous for this mutation, and one patient had a homozygous mutation, In comparison, 0 of 171 (0%) of sporadic breast cancer patients had the p53 13964(GC) mutation (P = 0.0003), We found that 0 of 42 (0%) of these hereditary breast cancer patients had other germ-line p53 mutation in exons 2-11, However, pedigree analysis demonstrated that all three patients had strong family histories of multiple types of cancers consistent with Li-Fraumeni syndrome but with late age of onset. Comprehensive BRCA1 and BRCA2 nucleotide analysis from patients with the p53 13964(GC) mutation revealed no concomitant deleterious BRCA1 or BRCA2 mutations, although they were found in the other hereditary breast cancer patients, Functional analysis of tno immortalized lymphoblastoid cell lines derived from patients with the p53 13964GC mutation demonstrated prolonged in vitro survival in response to cisplatinum treatment and showed decreased chemotherapy-induced apoptosis, Immunohistochemical analysis of breast tumors from these patients revealed high levels of mutant p53 protein, suggesting a functional mutation in the p53 gene. In summary, we have identified a single p53 intron mutation in familial breast cancer patients that is present at elevated frequency and has functional activity. C1 Thomas Jefferson Univ, Bodine Ctr Canc Treatment, Dept Radiat Oncol, Philadelphia, PA 19107 USA. Thomas Jefferson Univ, Kimmel Canc Ctr, Philadelphia, PA 19107 USA. Bioserve Biotechnol, Laurel, MD 20707 USA. Yale Univ, Sch Med, New Haven, CT 06520 USA. NCI, Human Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. RP Turner, BC (reprint author), Thomas Jefferson Univ, Bodine Ctr Canc Treatment, Dept Radiat Oncol, 111 S 11th St, Philadelphia, PA 19107 USA. RI Shields, Peter/I-1644-2012 NR 41 TC 64 Z9 69 U1 0 U2 3 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD FEB 15 PY 2000 VL 60 IS 4 BP 1062 EP 1069 PG 8 WC Oncology SC Oncology GA 287JQ UT WOS:000085503100048 PM 10706125 ER PT J AU Yuan, CJ Mandal, AK Zhang, ZJ Mukherjee, AB AF Yuan, CJ Mandal, AK Zhang, ZJ Mukherjee, AB TI Transcriptional regulation of Cyclooxygenase-2 gene expression: Novel effects of nonsteroidal anti-inflammatory drugs SO CANCER RESEARCH LA English DT Article ID PANCREATIC PHOSPHOLIPASE A(2); FAMILIAL ADENOMATOUS POLYPOSIS; MAMMARY EPITHELIAL-CELLS; 3 C/EBP ISOFORMS; COLORECTAL-CANCER; BINDING-SITES; NUCLEAR-ENVELOPE; COLON-CANCER; ASPIRIN USE; PROSTAGLANDIN AB Cyclooxygenase-2 (COX-2) gene overexpression is suggested to play important roles in colorectal tumorigenesis. Epidemiological studies revealed that nonsteroidal anti-inflammatory drugs (NSAIDs), such as aspirin and sulindac, which inhibit COX activity, reduce colorectal cancer mortality. Current investigations have focused on delineating the molecular mechanisms that regulate COX-2 gene expression and the roles of NSAIDs in cancer chemoprevention. COX-2 catalyzes the production of prostaglandins (PGs) from arachidonic acid (AA), generated by phospholipases A(2) (PLA(2)s), a family of acyl esterases that cause the release of AA from cellular phospholipids, Pancreatic secretory PLA(2) (sPLA(2)), via its receptor (sPLA(2)R), transcriptionally activates COX-2 gene expression in several cell types, although a specific transcription Factor mediating COX-2 expression has not yet been identified. Were, we report that a transcription factor, CCAAT/enhancer-binding protein beta (C/EBP beta), plays a critical role in sPLA(2)/IB-induced, receptor-mediated COX-2 gene expression in MC3T3E1 and NIH3T3 cells. Furthermore, treatment of these cells with NSAIDs in the presence of sPLA(2)/IB appears to potentiate the stimulatory effects on COX-2 mRNA and COX-2 protein expression and a concomitant elevation in PG production. Most significantly, NSAID treatment appears to drastically suppress the production of cytosolic PLA(2) (cPLA(2)) mRNA. The lack of sPLA(2)IB, sPLA(2)IIA, and sPLA(2)V mRNA expression in both NIH3T3 and MC3T3E1 cells suggests that cPLA(2) is the most likely enzyme that catalyzes the release of AA, the rate-limiting substrate of COX for the production of PGs. Our results suggest that: (a) sPLA,IB receptor-mediated COX-2 expression is mediated via C/EBP beta; (b) NSAIDs in the presence of sPLA(2)IB potentiate the stimulatory effects of sPLA(2)IB on COX-2 mRNA expression; and (c) despite the apparent stimulation of COX-2 expression by NSAIDs, they strikingly deprive COX-2 of its substrate, AA, by suppressing cPLA(2) mRNA expression. Both AA and PGs regulate many vital biological functions (e.g., motility and invasiveness) that are dysregulated in most cancer cells, and they have profound effects on cellular differentiation. Our results raise the possibility that deprivation of COX-2 of its substrate by the suppression of cPLA(2) mRNA expression is an additional mechanism used by NSAIDs to inhibit tumorigenesis. C1 NICHHD, Sect Dev Genet, Heritable Disorders Branch, NIH, Bethesda, MD 20892 USA. RP Mukherjee, AB (reprint author), NICHHD, Sect Dev Genet, Heritable Disorders Branch, NIH, Bldg 10,Room 9S241, Bethesda, MD 20892 USA. NR 68 TC 56 Z9 57 U1 0 U2 1 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD FEB 15 PY 2000 VL 60 IS 4 BP 1084 EP 1091 PG 8 WC Oncology SC Oncology GA 287JQ UT WOS:000085503100051 PM 10706128 ER PT J AU Chang, TH Szabo, E AF Chang, TH Szabo, E TI Induction of differentiation and apoptosis by ligands of peroxisome proliferator-activated receptor gamma in non-small cell lung cancer SO CANCER RESEARCH LA English DT Article ID CYCLIN D1 OVEREXPRESSION; HUMAN BREAST-CANCER; PPAR-GAMMA; TERMINAL DIFFERENTIATION; IN-VITRO; MONOCYTE/MACROPHAGE DIFFERENTIATION; GROWTH-INHIBITION; GENE-EXPRESSION; LEUKEMIA-CELLS; RETINOIC ACID AB The peroxisome proliferator-activated receptor gamma (PPAR gamma) is a ligand-activated transcription factor belonging to the steroid receptor superfamily. It is a key regulator of adipogenic differentiation, the ligands of which have also been demonstrated to induce differentiation in human breast and colon cancer cell lines. This study examined PPAR gamma in non-small cell lung cancer (NSCLC). PPAR gamma mRNA and protein were expressed in NSCLC cell lines, with highest levels in adenocarcinomas. PPAR gamma protein was also expressed in 50% of primary lung cancers by immunohistochemistry. Treatment of multiple cell lines with two distinct PPAR gamma Ligands in the presence of serum resulted in growth arrest, irreversible loss of capacity for anchorage-independent growth, decreased activity and expression of matrix metalloproteinase 2, and modulation of multiple markers in a manner consistent with differentiation. Specifically, there was up-regulation of general markers of the differentiated state such as gelsolin, Mad, and p21. Down-regulation of specific markers of progenitor lineages for the peripheral lung, i.e., the type II pneumocyte lineage markers MUC1 and surfactant protein-A and the Clara cell lineage marker CC10, also occurred. In addition, HTI56, a marker of terminally differentiated type I pneumocytes, was also induced. Consistent with a more mature, less malignant phenotype, ligand treatment also inhibited the expression of cyclin D1 and led to hypophosphorylation of the retinoblastoma protein. In contrast, in the absence of serum, ligand treatment rapidly resulted in apoptosis and substantially earlier onset of differentiation. Taken together, these results show that depending on the growth milieu, ligands of PPAR gamma induce differentiation and apoptosis in NSCLC, suggesting clinical utility for these agents. C1 NCI, Div Clin Sci, Med Branch, Cell & Canc Biol Dept, Rockville, MD 20850 USA. RP Szabo, E (reprint author), NCI, Div Canc Prevent, Lung & Aerodigest Canc Res Grp, 6130 Execut Blvd,Room 330, Bethesda, MD 20892 USA. NR 67 TC 335 Z9 353 U1 1 U2 6 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD FEB 15 PY 2000 VL 60 IS 4 BP 1129 EP 1138 PG 10 WC Oncology SC Oncology GA 287JQ UT WOS:000085503100058 PM 10706135 ER PT J AU Bazzano, LA He, J Ogden, LG Vupputuri, S Loria, C Myers, L Whelton, PK AF Bazzano, LA He, J Ogden, LG Vupputuri, S Loria, C Myers, L Whelton, PK TI Fruit and vegetable intake reduces cardiovascular disease mortality: Results from the NHANES I epidemiologic follow-up study (NHEFS) SO CIRCULATION LA English DT Meeting Abstract C1 Tulane Univ, Sch Publ Hlth & Trop Med, New Orleans, LA 70118 USA. NHLBI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 1 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA 5 BP 711 EP 711 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600027 ER PT J AU Ogg, MS Tracy, RP Luong, L Kuller, LH Arnold, AM Sharrett, AR Humphries, S AF Ogg, MS Tracy, RP Luong, L Kuller, LH Arnold, AM Sharrett, AR Humphries, S TI The-174 G/C interleukin-6 polymorphism is associated with increased levels of inflammation markers and increased blood pressure. SO CIRCULATION LA English DT Meeting Abstract C1 UCL, London WC1E 6BT, England. Univ Vermont, Colchester, VT USA. Univ Pittsburgh, Pittsburgh, PA 15260 USA. Univ Washington, Seattle, WA 98195 USA. NHLBI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA 7 BP 712 EP 712 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600029 ER PT J AU O'Donnell, CJ Larson, MG Levy, D Benjamin, E Manning, WJ D'Agostino, RA Clouse, ME AF O'Donnell, CJ Larson, MG Levy, D Benjamin, E Manning, WJ D'Agostino, RA Clouse, ME TI Relationship of C-reactive protein to coronary and aortic calcification in the framingham heart study SO CIRCULATION LA English DT Meeting Abstract C1 NHLBI, Framingham Heart Study, Framingham, MA USA. Boston Univ, Boston, MA 02215 USA. Beth Israel Deaconess Med Ctr, Boston, MA USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA 15 BP 713 EP 714 PG 2 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600037 ER PT J AU Feldman, AH Kuller, LH Sutton-Tyrrell, K Lumley, T Manolio, TA Burke, GL Furberg, CD O'Leary, D AF Feldman, AH Kuller, LH Sutton-Tyrrell, K Lumley, T Manolio, TA Burke, GL Furberg, CD O'Leary, D TI Assessing risk and prognostic value of various subclinical measures stratified by carotid intimal medial thickness quintiles SO CIRCULATION LA English DT Meeting Abstract C1 Univ Pittsburgh, Pittsburgh, PA 15260 USA. Univ Washington, Seattle, WA 98195 USA. NHLBI, Washington, DC USA. Wake Forest Univ, Sch Med, Winston Salem, NC 27109 USA. New England Med Ctr, Boston, MA 02111 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P1 BP 717 EP 717 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600049 ER PT J AU Newman, AB Arnold, AM Burke, GL O'Leary, D Manolio, TA AF Newman, AB Arnold, AM Burke, GL O'Leary, D Manolio, TA TI Abdominal aortic aneurysm in older adults predicts total mortality and incident cardiovascular disease: The cardiovascular health study SO CIRCULATION LA English DT Meeting Abstract C1 Univ Pittsburgh, Pittsburgh, PA 15260 USA. Univ Washington, Seattle, WA 98195 USA. Wake Forest Univ, Sch Med, Winston Salem, NC 27109 USA. New England Med Ctr, Boston, MA 02111 USA. NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P12 BP 719 EP 719 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600060 ER PT J AU Jacobs, DR Tracy, RP Gross, MD Loria, C Bild, D Liu, K McCreath, H AF Jacobs, DR Tracy, RP Gross, MD Loria, C Bild, D Liu, K McCreath, H TI Plasma C-reactive protein and vitamin C are associated with coronary calcification in young adults: The CARDIA study SO CIRCULATION LA English DT Meeting Abstract C1 Univ Minnesota, Minneapolis, MN 55455 USA. Univ Vermont, Colchester, VT USA. DECA, NIH, NHLBI, Bethesda, MD USA. Northwestern Univ, Chicago, IL 60611 USA. Univ Alabama, Birmingham, AL USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P21 BP 720 EP 721 PG 2 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600069 ER PT J AU Wu, TJ Dorn, JP Donahue, RP Sempos, CT AF Wu, TJ Dorn, JP Donahue, RP Sempos, CT TI Serum C-reactive protein elevation associated with higher levels of plasma glucose and serum insulin: A study of a national sample of US adults SO CIRCULATION LA English DT Meeting Abstract C1 SUNY Buffalo, Buffalo, NY 14260 USA. NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P20 BP 720 EP 720 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600068 ER PT J AU Liao, DP Pankow, JS Sharrett, R Cascio, W Evans, GW Cai, JW AF Liao, DP Pankow, JS Sharrett, R Cascio, W Evans, GW Cai, JW TI Inflammation is associated with impaired autonomic cardiac control-the ARIC study. SO CIRCULATION LA English DT Meeting Abstract C1 Penn State Univ, Coll Med, Hershey, PA 17033 USA. Univ N Carolina, Chapel Hill, NC 27515 USA. NHLBI, NIH, Bethesda, MD 20892 USA. Wake Forest Univ, Sch Med, Winston Salem, NC 27109 USA. NR 0 TC 0 Z9 0 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P23 BP 721 EP 721 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600071 ER PT J AU Djousse, L Levy, D Murabito, JM Cupples, LA Ellison, RC AF Djousse, L Levy, D Murabito, JM Cupples, LA Ellison, RC TI Alcohol consumption and risk of intermittent claudication in the Framingham Heart Study SO CIRCULATION LA English DT Meeting Abstract C1 Boston Univ, Sch Med, Boston, MA 02215 USA. NHLBI, Framingham Heart Study, Framingham, MA USA. Boston Univ, Sch Publ Hlth, Boston, MA 02215 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P32 BP 723 EP 723 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600080 ER PT J AU Merz, CNB Johnson, BD Sharaf, BL Pepine, CJ Reichek, N Reis, SE Rogers, WJ Sopko, G Kelsey, SF AF Merz, CNB Johnson, BD Sharaf, BL Pepine, CJ Reichek, N Reis, SE Rogers, WJ Sopko, G Kelsey, SF TI Age, risk factors and coronary artery disease severity in women: The NHLBI-sponsored WISE study SO CIRCULATION LA English DT Meeting Abstract C1 Cedars Sinai Med Ctr, Los Angeles, CA 90048 USA. Univ Pittsburgh, Pittsburgh, PA 15260 USA. Brown Univ, Providence, RI 02912 USA. Univ Florida, Gainesville, FL 32611 USA. Allegheny Gen Hosp, Pittsburgh, PA 15212 USA. Univ Alabama, Birmingham, AL USA. NHLBI, Bethesda, MD 20892 USA. RI Reis, Steven/J-3957-2014 NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P51 BP 726 EP 727 PG 2 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600099 ER PT J AU Sutton-Tyrrell, K Havlik, RJ Vaitkevicius, PV Spurgeon, HA Pahor, M Simonsick, EM Lakatta, E Newman, AB AF Sutton-Tyrrell, K Havlik, RJ Vaitkevicius, PV Spurgeon, HA Pahor, M Simonsick, EM Lakatta, E Newman, AB TI Greater aortic stiffness among blacks compared to whites in the study of health, aging and body composition (Health ABC) SO CIRCULATION LA English DT Meeting Abstract C1 Univ Pittsburgh, Pittsburgh, PA 15260 USA. NIA, Bethesda, MD 20892 USA. Johns Hopkins Univ, Baltimore, MD 21218 USA. Univ Tennessee, Memphis, TN 38163 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P47 BP 726 EP 726 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600095 ER PT J AU Bairey, CN McGorray, SP Sharaf, BL Pepine, CJ Reichek, N Reis, SE Rogers, WJ Sopko, G Kelsey, SF AF Bairey, CN McGorray, SP Sharaf, BL Pepine, CJ Reichek, N Reis, SE Rogers, WJ Sopko, G Kelsey, SF TI Race, risk factors and coronary artery disease in women: The NHLBI-Sponsored WISE study SO CIRCULATION LA English DT Meeting Abstract C1 Cedars Sinai Med Ctr, Los Angeles, CA 90048 USA. Univ Florida, Gainesville, FL 32611 USA. Brown Univ, Providence, RI 02912 USA. Allegheny Gen Hosp, Pittsburgh, PA 15212 USA. Univ Alabama, Birmingham, AL USA. NHLBI, Bethesda, MD 20892 USA. RI Reis, Steven/J-3957-2014 NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P52 BP 727 EP 727 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600100 ER PT J AU MacCluer, JW Robbins, DC Lee, ET Welty, TK Fabsitz, RR Howard, BV AF MacCluer, JW Robbins, DC Lee, ET Welty, TK Fabsitz, RR Howard, BV TI Heritabilities of cardiovascular disease risk factors in American indians: The strong heart study SO CIRCULATION LA English DT Meeting Abstract C1 SW Fdn Biomed Res, San Antonio, TX 78284 USA. Medstar Res Inst, Washington, DC USA. Univ Oklahoma, Oklahoma City, OK USA. Aberdeen Area Tribal Chairmens Hlth Board, Rapid City, SD USA. NHLBI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P57 BP 728 EP 728 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600105 ER PT J AU Rodriguez, BL Resnick, H Curb, JD Chen, R Michele-Baraquino, N Foley, D Masaki, KH Schatz, I Petrovitch, H Ross, W Abbott, RD AF Rodriguez, BL Resnick, H Curb, JD Chen, R Michele-Baraquino, N Foley, D Masaki, KH Schatz, I Petrovitch, H Ross, W Abbott, RD TI ADA and WHO classifications for diabetes: Their impact on diabetes prevalence, total and CHD mortality in elderly Japanese-American men SO CIRCULATION LA English DT Meeting Abstract C1 Univ Hawaii, Honolulu, HI 96822 USA. NIA, Bethesda, MD 20892 USA. Univ Virginia, Charlottesville, VA USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P61 BP 729 EP 729 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600109 ER PT J AU D'Agostino, RB Ehrlich, J Schoentgen, S Savage, PJ Rewers, M AF D'Agostino, RB Ehrlich, J Schoentgen, S Savage, PJ Rewers, M TI Insulin resistance predicts coronary artery calcification independent of CVD risk factors or diabetes status SO CIRCULATION LA English DT Meeting Abstract C1 Wake Forest Univ, Bowman Gray Sch Med, Winston Salem, NC USA. Univ Colorado, Denver, CO 80202 USA. Univ Iowa, Iowa City, IA USA. NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P68 BP 730 EP 730 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600116 ER PT J AU Haffner, SM D'Agostino, RB Wagenknecht, LE Savage, PJ AF Haffner, SM D'Agostino, RB Wagenknecht, LE Savage, PJ TI Increased carotid wall thickness prior to the onset of type 2 diabetes SO CIRCULATION LA English DT Meeting Abstract C1 Univ Texas, Hlth Sci Ctr, San Antonio, TX USA. Wake Forest Univ, Bowman Gray Sch Med, Winston Salem, NC USA. NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P66 BP 730 EP 730 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600114 ER PT J AU Lloyd-Jones, DM Larson, MG Beiser, A D'Agostino, RB Wilson, PW Cleeman, JI Levy, D AF Lloyd-Jones, DM Larson, MG Beiser, A D'Agostino, RB Wilson, PW Cleeman, JI Levy, D TI Lifetime risk of coronary heart disease by total cholesterol level SO CIRCULATION LA English DT Meeting Abstract C1 NHLBI, Framingham Heart Study, Framingham, MA USA. Boston Univ, Boston, MA 02215 USA. NHLBI, Natl Cholesterol Educ Program, Bethesda, MD 20892 USA. RI Lloyd-Jones, Donald/C-5899-2009 NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P78 BP 732 EP 732 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600126 ER PT J AU Schreiner, PJ Jacobs, DR Folsom, AR Hulley, SB AF Schreiner, PJ Jacobs, DR Folsom, AR Hulley, SB TI Apolipoprotein B versus LDL-cholesterol as predictors of coronary artery calcification in young adults: The CARDIA study SO CIRCULATION LA English DT Meeting Abstract C1 Univ Minnesota, Minneapolis, MN USA. NHLBI, NIH, Bethesda, MD 20892 USA. Univ Calif San Francisco, San Francisco, CA 94143 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P76 BP 732 EP 732 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600124 ER PT J AU Bazzano, LA He, J Ogden, LG Vupputuri, S Loria, C Myers, L Whelton, PK AF Bazzano, LA He, J Ogden, LG Vupputuri, S Loria, C Myers, L Whelton, PK TI Bean consumption and incidence of coronary heart disease in US men and women: NHANES I epidemiologic follow-up study SO CIRCULATION LA English DT Meeting Abstract C1 Tulane Univ, Sch Publ Hlth & Trop Med, New Orleans, LA USA. NHLBI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P89 BP 734 EP 734 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600137 ER PT J AU Gardner, CD Dixon, LB Winkleby, MA AF Gardner, CD Dixon, LB Winkleby, MA TI Homocysteine levels, vitamin intake and dietary supplement use among black, Mexican-American and white US adults: The third national health and nutrition examination survey, (NHANES III, phase 2, 1991-1994) SO CIRCULATION LA English DT Meeting Abstract C1 Stanford Univ, Palo Alto, CA 94304 USA. NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 15 PY 2000 VL 101 IS 6 MA P86 BP 734 EP 734 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 284NZ UT WOS:000085339600134 ER PT J AU Qian, XL Esteban, L Vass, WC Upadhyaya, C Papageorge, AG Yienger, K Ward, JM Lowy, DR Santos, E AF Qian, XL Esteban, L Vass, WC Upadhyaya, C Papageorge, AG Yienger, K Ward, JM Lowy, DR Santos, E TI The Sos1 and Sos2 Ras-specific exchange factors: differences in placental expression and signaling properties SO EMBO JOURNAL LA English DT Article DE ERK activation pathway; genomic disruption; placental development; Ras signaling; spongiotrophoblasts ID EPIDERMAL GROWTH-FACTOR; GTPASE-ACTIVATING PROTEIN; KINASE PHOSPHORYLATION; TYROSINE KINASE; MICE LACKING; EGF RECEPTOR; V-SRC; TRANSFORMATION; IDENTIFICATION; MOUSE AB Targeted disruption of both alleles of mouse sos1, which encodes a Ras-specific exchange factor, conferred mid-gestational embryonic lethality that was secondary to impaired placental development and was associated,vith very low placental ERK activity. The trophoblastic layers of sos1(-/-) embryos were poorly developed, correlating with high sos1 expression in wild-type trophoblasts. A sos1(-/-) cell line, which expressed readily detectable levels of the closely related Sos2 protein, formed complexes between Sos2, epidermal growth factor receptor (EGFR) and Shc efficiently, gave normal Ras GTP and ERK responses when treated with EGF for less than or equal to 10 min and was transformed readily by activated Ras, However, the sos1(-/-) cells were resistant to transformation by v-Src or by overexpressed EGFR and continuous EGF treatment, unlike sos1(+/-) or wild-type cells. This correlated with Sos2 binding less efficiently than Sos1 to EGFR and Shc in cells treated with EGF for greater than or equal to 90 min or to v-Src and Shc in v-Src-expressing cells, acid with less ERK activity. We conclude that Sos1 participates in both short- and long-term signaling, while Sos2-dependent signals are predominantly short-term. C1 NCI, Cellular Oncol Lab, Bethesda, MD 20892 USA. NCI, Lab Cellular & Mol Biol, Bethesda, MD 20892 USA. NCI, Vet Tumor Pathol Sect, Frederick, MD 21702 USA. RP Lowy, DR (reprint author), NCI, Cellular Oncol Lab, Bethesda, MD 20892 USA. NR 57 TC 66 Z9 68 U1 0 U2 2 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD FEB 15 PY 2000 VL 19 IS 4 BP 642 EP 654 DI 10.1093/emboj/19.4.642 PG 13 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 285NJ UT WOS:000085394900017 PM 10675333 ER PT J AU Vogel, JL Kristie, TM AF Vogel, JL Kristie, TM TI The novel coactivator C1 (HCF) coordinates multiprotein enhancer formation and mediates transcription activation by GABP SO EMBO JOURNAL LA English DT Article DE coactivator; herpes simplex virus; protein interactions; transcription ID OCT-1 HOMEO DOMAIN; BINDING PROTEIN; TRANSACTIVATOR VP16; CELLULAR FACTOR; GENE-TRANSCRIPTION; DNA COMPLEX; POU DOMAIN; SIMPLEX; PROMOTER; ETS AB Transcription of the herpes simplex virus 1 (HSV-1) immediate early (IE) genes is determined by multiprotein enhancer complexes. The core enhancer assembly requires the interactions of the POU-homeodomain protein Oct-1, the viral transactivator alpha TIF and the cellular factor C1 (HCF). In this context, the C1 factor interacts with each protein to assemble the stable enhancer complex. In addition, the IE enhancer cores contain adjacent binding sites for other cellular transcription factors such as Sp1 and GA-binding protein (GABP). In this study, a direct interaction of the C1 factor with GABP is demonstrated, defining the C1 factor as the critical coordinator of the enhancer complex assembly. In addition, mutations that reduce the GABP transactivation potential also impair the C1-GABP interaction, indicating that the C1 factor functions as a novel coactivator of GABP-mediated transcription. The interaction and coordinated assembly of the enhancer proteins by the C1 factor may be critical for the regulation of the HSV lytic-latent cycle. C1 NIH, Viral Dis Lab, Bethesda, MD 20892 USA. RP Kristie, TM (reprint author), NIH, Viral Dis Lab, Bldg 4,Room 133,4 Ctr Dr, Bethesda, MD 20892 USA. NR 54 TC 58 Z9 62 U1 0 U2 1 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD FEB 15 PY 2000 VL 19 IS 4 BP 683 EP 690 DI 10.1093/emboj/19.4.683 PG 8 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 285NJ UT WOS:000085394900021 PM 10675337 ER PT J AU Jin, DY Wang, HL Zhou, Y Chun, ACS Kibler, KV Hou, YD Kung, HF Jeang, KT AF Jin, DY Wang, HL Zhou, Y Chun, ACS Kibler, KV Hou, YD Kung, HF Jeang, KT TI Hepatitis C virus core protein-induced loss of LZIP function correlates with cellular transformation SO EMBO JOURNAL LA English DT Article DE bZIP transcription factor; hepatitis C virus; hepatitis C virus core protein; hepatocellular carcinoma; LZIP ID LYMPHOTOXIN-BETA RECEPTOR; RAT EMBRYO FIBROBLASTS; HEPATOCELLULAR-CARCINOMA; TRANSCRIPTION FACTORS; SUBCELLULAR-LOCALIZATION; NUCLEAR-LOCALIZATION; PROMOTER ACTIVITY; TRANSGENIC MICE; EXPRESSION; APOPTOSIS AB Hepatitis C virus (HCV) is the major etiological agent of blood-borne non-A non-B hepatitis and a leading cause of liver cirrhosis and hepatocellular carcinoma worldwide. HCV core protein is a multifunctional protein with regulatory functions in cellular transcription and virus-induced transformation and pathogenesis. Here we report on the identification of a bZIP nuclear transcription protein as an HCV core cofactor for transformation. This bZIP factor, designated LZIP, activates CRE-dependent transcription and regulates cell proliferation. Loss of LZIP function in NIH 3T3 cells triggers morphological transformation and anchorage-independent growth. We show that HCV core protein aberrantly sequesters LZIP in the cytoplasm, inactivates LZIP function and potentiates cellular transformation. Our findings suggest that LZIP might serve a novel cellular tumor suppressor function that is targeted by the HCV core. C1 NIAID, Mol Microbiol Lab, NIH, Bethesda, MD 20892 USA. Chinese Acad Prevent Med, Inst Virol, Natl Key Lab Mol Virol & Genet Engn, Beijing 100052, Peoples R China. Univ Hong Kong, Inst Mol Biol, Hong Kong, Hong Kong, Peoples R China. RP Jin, DY (reprint author), NIAID, Mol Microbiol Lab, NIH, Bethesda, MD 20892 USA. RI Jeang, Kuan-Teh/A-2424-2008 NR 88 TC 111 Z9 126 U1 0 U2 1 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0261-4189 J9 EMBO J JI Embo J. PD FEB 15 PY 2000 VL 19 IS 4 BP 729 EP 740 DI 10.1093/emboj/19.4.729 PG 12 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 285NJ UT WOS:000085394900026 PM 10675342 ER PT J AU Galperin, MY Aravind, L Koonin, EV AF Galperin, MY Aravind, L Koonin, EV TI Aldolases of the DhnA family: a possible solution to the problem of pentose and hexose biosynthesis in archaea SO FEMS MICROBIOLOGY LETTERS LA English DT Article DE glycolysis; gluconeogenesis; ribose; fructose bisphosphate aldolase; deoxyribose phosphate aldolase; Chlamydia trachomatis ID COMPLETE GENOME SEQUENCE; AUTOTROPH METHANOBACTERIUM-THERMOAUTOTROPHICUM; FRUCTOSE-1,6-BISPHOSPHATE ALDOLASE; METHANOCOCCUS-MARIPALUDIS; RIBOSE BIOSYNTHESIS; ACTIVE-SITE; GENE; PATHWAY; JANNASCHII; METABOLISM AB Sequence analysis of the recently identified class I aldolase of Escherichia coli (dhnA gene product) helped to identify its homologs in Chlamydia trachomatis, Chlamydiophyla pneumoniae and in each of the completely sequenced archaeal genomes. Iterative database searches revealed sequence similarities between the DhnA-family enzymes, deoxyribose phosphate aldolases and bacterial (class II) fructose bisphosphate aldolases and allowed prediction of similar three-dimensional structures (TIM-barrel fold) in all these enzymes. The Schiff base-forming lysyl residues of DhnA and deoxyribose phosphate aldolase are conserved in all members of the DhnA and deoxyribose phosphate aldolase families, indicating that these enzymes share common features with both class I and class II aldolases. The DhnA-family enzymes are predicted to possess an aldolase activity and to play a critical role in sugar biosynthesis in archaea. (C) 2000 Published by Elsevier Science B.V. All rights reserved. C1 NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA. Texas A&M Univ, Dept Biol, College Stn, TX 77843 USA. RP Galperin, MY (reprint author), NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA. OI Galperin, Michael/0000-0002-2265-5572 NR 44 TC 31 Z9 31 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-1097 J9 FEMS MICROBIOL LETT JI FEMS Microbiol. Lett. PD FEB 15 PY 2000 VL 183 IS 2 BP 259 EP 264 DI 10.1111/j.1574-6968.2000.tb08968.x PG 6 WC Microbiology SC Microbiology GA 284UX UT WOS:000085352200011 PM 10675594 ER PT J AU Carpten, JD Makalowska, I Robbins, CM Scott, N Sood, R Connors, TD Bonner, TI Smith, JR Faruque, MU Stephan, DA Pinkett, H Morgenbesser, SD Su, K Graham, C Gregory, SG Williams, H McDonald, L Baxevanis, AD Klingler, KW Landes, GM Trent, JM AF Carpten, JD Makalowska, I Robbins, CM Scott, N Sood, R Connors, TD Bonner, TI Smith, JR Faruque, MU Stephan, DA Pinkett, H Morgenbesser, SD Su, K Graham, C Gregory, SG Williams, H McDonald, L Baxevanis, AD Klingler, KW Landes, GM Trent, JM TI A 6-Mb high-resolution physical and transcription map encompassing the hereditary prostate cancer 1 (HPC1) region SO GENOMICS LA English DT Article ID SUSCEPTIBILITY LOCUS; HUMAN GENOME; HUMAN DNA; FAMILIAL ARTHROPATHY; CHROMOSOME 1Q; CPG ISLANDS; COXA VARA; GENE; HYPERPARATHYROIDISM; PERICARDITIS AB Several hereditary disease loci have been genetically mapped to the chromosome 1q24-q31 interval, including the hereditary prostate cancer 1 (HPC1) locus. Here, we report the construction of a 20-Mb yeast artificial chromosome contig and a high-resolution 6-Mb sequence-ready bacterial artificial chromosome (BAC)/P1-derived artificial chromosome (PAC) contig of 1q25 by sequence and computational analysis, STS content mapping, and chromosome walking. One hundred thirty-six new STSs, including 10 novel simple sequence repeat polymorphisms that are being used for genetic refinement of multiple disease loci, have been generated from this contig and are shown to map to the 1q25 interval. The integrity of the 6-Mb BAC/PAC contig has been confirmed by restriction fingerprinting, and this contig is being used as a template for human chromosome 1 genome sequencing. A transcription mapping effort has resulted in the precise localization of 18 known genes and 31 ESTs by database searching, exon trapping, direct cDNA hybridization, and sample sequencing of BACs from the 1q25 contig, An additional 11 known genes and ESTs have been placed within the larger 1q24-q31 interval. These transcription units represent candidate genes for multiple hereditary diseases, including HPC1. (C) 2000 Academic Press. C1 NHGRI, NIH, Canc Genet Branch, Bethesda, MD 20892 USA. NHGRI, NIH, Gene Technol Branch, Bethesda, MD 20892 USA. NIMH, NIH, Bethesda, MD 20892 USA. Genzyme Genet Corp, Framingham, MA 01701 USA. Sanger Ctr, Cambridge, England. RP Carpten, JD (reprint author), NHGRI, NIH, Canc Genet Branch, Bldg 36,Room 3D04,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Smith, Jeff/C-3484-2012; Pinkett, Heather/M-9235-2014 NR 42 TC 24 Z9 24 U1 0 U2 1 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0888-7543 J9 GENOMICS JI Genomics PD FEB 15 PY 2000 VL 64 IS 1 BP 1 EP 14 DI 10.1006/geno.1999.6051 PG 14 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA 295VH UT WOS:000085988200001 PM 10708513 ER PT J AU Schriml, LM Dean, M AF Schriml, LM Dean, M TI Identification of 18 mouse ABC genes and characterization of the ABC superfamily in Mus musculus SO GENOMICS LA English DT Article ID MULTIDRUG-RESISTANCE; ANTIGEN PRESENTATION; LIVER; MICE; TRANSPORTER; SECRETION; PROTEINS; CLONING; DISEASE; DNA AB ATP-binding cassette (ABC) genes encode a family of transport proteins known to be involved in a number of human genetic diseases. In this study, we characterized the ABC superfamily in Mus musculus through in silico gene identification and mapping and phylogenetic analysis of mouse and human ABC genes. By querying dbEST with amino acid sequences from the conserved ATP-binding domains, we identified and partially sequenced 18 new mouse ABC genes, bringing the total number of mouse ABC genes to 34. Twelve of the new ABC genes were mapped in the mouse genome to the X chromosome and to 10 of the 19 autosomes. Phylogenetic relationships of mouse and human ABC genes were examined with maximum parsimony and neighbor-joining analyses that demonstrated that mouse and human ABC orthologs are more closely related than are mouse paralogs. The mouse ABC genes could be grouped into the seven previously described human ABC subfamilies. Three mouse ABC genes mapped to regions implicated in cholesterol gallstone susceptibility. (C) 2000 Academic Press. C1 NCI, Frederick Canc Res & Dev Ctr, Human Genet Sect, Lab Genom Divers, Frederick, MD 21702 USA. RP Dean, M (reprint author), NCI, Frederick Canc Res & Dev Ctr, Human Genet Sect, Lab Genom Divers, Bldg 560,Room 21-18, Frederick, MD 21702 USA. RI Dean, Michael/G-8172-2012; OI Dean, Michael/0000-0003-2234-0631; Schriml, Lynn/0000-0001-8910-9851 FU NCI NIH HHS [N01-CO-56000] NR 33 TC 26 Z9 30 U1 1 U2 1 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0888-7543 J9 GENOMICS JI Genomics PD FEB 15 PY 2000 VL 64 IS 1 BP 24 EP 31 DI 10.1006/geno.1999.6102 PG 8 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA 295VH UT WOS:000085988200003 PM 10708515 ER PT J AU Bliskovski, V Liddell, R Ramsay, ES Miller, MJ Mock, BA AF Bliskovski, V Liddell, R Ramsay, ES Miller, MJ Mock, BA TI Structure and localization of mouse PMSCL1 and PMSCL2 genes SO GENOMICS LA English DT Article ID PM-SCL; DIABETES SUSCEPTIBILITY; AUTOIMMUNE-DISEASE; NUCLEOLAR ANTIGEN; OVERLAP SYNDROMES; CYCLIN-A; AUTOANTIGEN; PROTEIN; REGION; MICE AB Sera from some patients with polymyositis-scleoderma overlap syndrome (PRI-SCL) recognize two antigenically unrelated proteins, PMSCL1 and PMSCL2. Complete mouse Pmscl1 and Pmscl2 cDNA sequences, chromosomal localizations, exon/intron structure, and promoter region sequences of the mouse Pmscl2 gene are presented. The PMSCL1 gene was found to overlap significantly with cyclin A2 in both human and mouse. As such, it may be deduced that PMSCL1 sequences map to human chromosome 4q27 and the proximal portion of mouse chromosome (Chr) 3 where human and mouse cyclin A2 genes reside. Analysis of human and mouse PMSCL1 cDNA sequences provides evidence that the PMSCL1 protein is 68 amino acids longer than previously thought. A BAC containing mouse Pmscl2 was localized to distal mouse Chr 4 by FISH. This BAC contains the microsatellite D4Mit310. D4Mit310 colocalizes with a number of genes that map to human 1p36. In fact, a STS (G25404) located 54.6 cR from the top of human chromosome 1 was found to contain PMSCL2 sequence upon BLAST search. (C) 2000 Academic Press. C1 NCI, NIH, Genet Lab, DBS, Bethesda, MD 20892 USA. NCI, NIH, Expt Carcinogenesis Lab, DBS, Bethesda, MD 20892 USA. RP Mock, BA (reprint author), NCI, NIH, Genet Lab, DBS, Bldg 37,Room 2B08,37 Convent Dr,MSC 4255, Bethesda, MD 20892 USA. NR 20 TC 3 Z9 5 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0888-7543 J9 GENOMICS JI Genomics PD FEB 15 PY 2000 VL 64 IS 1 BP 106 EP 110 DI 10.1006/geno.2000.6118 PG 5 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA 295VH UT WOS:000085988200012 PM 10708524 ER PT J AU Freund, YR Mirsalis, JC Fairchild, DG Brune, J Hokama, LA Schindler-Horvat, J Tomaszewski, JE Hodge, JW Schlom, J Kantor, JA Tyson, CA Donohue, SJ AF Freund, YR Mirsalis, JC Fairchild, DG Brune, J Hokama, LA Schindler-Horvat, J Tomaszewski, JE Hodge, JW Schlom, J Kantor, JA Tyson, CA Donohue, SJ TI Vaccination with a recombinant vaccinia vaccine containing the B7-1 co-stimulatory molecule causes no significant toxicity and enhances T cell-mediated cytotoxicity SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article ID PRIMARY BILIARY-CIRRHOSIS; ANTITUMOR IMMUNITY; ANTIMITOCHONDRIAL ANTIBODY; VIRUS-VACCINE; ANTIGEN; COSTIMULATION; MICE; CEA; IMMUNIZATION; ADMIXTURE AB B7-1 is a co-stimulatory molecule that provides a second signal for T-cell activation. Several studies have demonstrated that vaccination with a vector containing genes encoding B7-1 and an antigen appears to be efficacious at promoting immune responsiveness to the antigen, To evaluate the safety of such a protocol and determine the effect of the B7-1 vector on immune responsiveness, female C57BL/6 mice were administered Wyeth wild-type vaccinia virus (V-WT) or V-WT containing the gene for B7-1 (rV-B7-1) as a single s.c. injection or 3 monthly s.c. injections. Immunologic parameters were evaluated in half of the mice and general toxicity in the other half. Immunologic end points included determination of splenic lymphocyte phenotypes, mitogen-induced T- and B-cell proliferation, T-cell proliferation in response to alloantigens, cell-mediated cytotoxicity (CMC), natural killer cell activity and serum anti-nuclear antibody (ANA) titers, No significant signs of general toxicity were noted. The primary immunologic effect was an increase in the ability of spleen cells to lyse allogeneic targets and to proliferate in response to allogeneic stimulation. Numbers of splenic CD8(+) cells were also increased. These effects were more pronounced after 3 vaccinations than after a single vaccination. Minimal differences in ANA were observed between mice immunized with V-WT and rV-B7-1, In addition, no serum antibodies against B7-1 were detected in any mice. The data suggest that vaccination with rV-B7-1 augments CMC with minimal toxicity. Int. J, Cancer 85:508-517, 2000, (C) 2000 Wiley-Liss, Inc. C1 SRI Int, Menlo Park, CA 94025 USA. NCI, Toxicol & Pharmacol Branch, Bethesda, MD 20892 USA. NCI, Tumor Immunol & Biol Lab, Bethesda, MD 20892 USA. RP Freund, YR (reprint author), SRI Int, 333 Ravenswood Ave, Menlo Park, CA 94025 USA. RI Hodge, James/D-5518-2015 OI Hodge, James/0000-0001-5282-3154 FU NCI NIH HHS [N01-CM-37837]; PHS HHS [87101] NR 22 TC 13 Z9 13 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD FEB 15 PY 2000 VL 85 IS 4 BP 508 EP 517 DI 10.1002/(SICI)1097-0215(20000215)85:4<508::AID-IJC11>3.0.CO;2-D PG 10 WC Oncology SC Oncology GA 280WX UT WOS:000085127700011 PM 10699923 ER PT J AU Franchimont, D Galon, J Gadina, M Visconti, R Zhou, YJ Aringer, M Frucht, DM Chrousos, GP O'Shea, JJ AF Franchimont, D Galon, J Gadina, M Visconti, R Zhou, YJ Aringer, M Frucht, DM Chrousos, GP O'Shea, JJ TI Inhibition of Th1 immune response by glucocorticoids, dexamethasone selectively inhibits IL-12-induced Stat4 phosphorylation in T lymphocytes SO JOURNAL OF IMMUNOLOGY LA English DT Article ID EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS; TYROSINE PHOSPHORYLATION; SH2-CONTAINING PROTEIN; SIGNAL-TRANSDUCTION; CD4(+) LYMPHOCYTES; IL-12 PRODUCTION; LEWIS RATS; CELLS; ACTIVATION; RECEPTOR AB Glucocorticoids are widely used in the therapy of inflammatory, autoimmune, and allergic diseases. As the end-effecters of the hypothalamic-pituitary-adrenal axis, endogenous glucocorticoids also play an important role in suppressing innate and cellular immune responses. Previous studies have indicated that glucocorticoids inhibit Th1 and enhance Th2 cytokine secretion. IL-12 promotes Th1 cell-mediated immunity, while IL-4 stimulates Th2 humoral-mediated immunity. Here, we examined the regulatory effect of glucocorticoids on key elements of IL-12 and IL-4 signaling. We first investigated the effect of dexamethasone on IL-12-inducible genes and showed that dexamethasone inhibited IL-12-induced IFN-gamma secretion and IFN regulatory factor-1 expression in both NK and T cells. This occurred even though the level of expression of IL-12 receptors and IL-12-induced Janus kinase phosphorylation remained unaltered. However, dexamethasone markedly inhibited IL-12-induced phosphorylation of Stat4 without altering its expression. This was specific, as IL-4-induced Stat6 phosphorylation was not affected, and mediated by the glucocorticoid receptor, as it was antagonized by the glucocorticoid receptor antagonist RU486, Moreover, transfection experiments showed that dexamethasone reduced responsiveness to IL-12 through the inhibition of Stat4-dependent IPN regulatory factor-1 promoter activity. We conclude that blocking IL-12-induced Stat4 phosphorylation, without altering IL-4-induced Stat6 phosphorylation, appears to be a new suppressive action of glucocorticoids on the Th1 cellular immune response and may help explain the glucocorticoid-induced shift toward the Th2 humoral immune response. C1 NIAMSD, Lymphocyte Cell Biol Sect, NIH, Bethesda, MD 20892 USA. NICHHD, Dev Endocrinol Branch, NIH, Bethesda, MD 20892 USA. RP O'Shea, JJ (reprint author), NIAMSD, Lymphocyte Cell Biol Sect, NIH, Bldg 10,Room 10N262,10 Ctr Dr, Bethesda, MD 20892 USA. RI Visconti, Roberta/C-5299-2009 NR 45 TC 151 Z9 156 U1 2 U2 4 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD FEB 15 PY 2000 VL 164 IS 4 BP 1768 EP 1774 PG 7 WC Immunology SC Immunology GA 283VK UT WOS:000085296600020 PM 10657623 ER PT J AU Moratz, C Kang, VH Druey, KM Shi, CS Scheschonka, A Murphy, PM Kozasa, T Kehrl, JH AF Moratz, C Kang, VH Druey, KM Shi, CS Scheschonka, A Murphy, PM Kozasa, T Kehrl, JH TI Regulator of G protein signaling 1 (RGS1) markedly impairs G(i alpha) signaling responses of B lymphocytes SO JOURNAL OF IMMUNOLOGY LA English DT Article ID PLATELET-ACTIVATING-FACTOR; CHEMOKINE RECEPTOR CXCR4; NF-KAPPA-B; PERTUSSIS TOXIN; ALPHA-SUBUNITS; CELL-LINE; COUPLED RECEPTORS; TRANSITION-STATE; FAMILY MEMBERS; P115 RHOGEF AB Regulator of G protein signaling (RGS) proteins modulate signaling through pathways that use heterotrimeric G proteins as transducing elements. RGS1 is expressed at high levels in certain B cell lines and can be induced in normal B cells by treatment with TNF-alpha. To determine the signaling pathways that RGS1 may regulate, we examined the specificity of RGS1 for various G, subunits and assessed its effect on chemokine signaling. G protein binding and GTPase assays revealed that RGS1 is a G,, and G(q alpha) GTPase-activating protein and a potential G(12 alpha) effector antagonist, Functional studies demonstrated that RGS1 impairs platelet activating factor-mediated increases in intracellular Ca+2, stromal-derived factor-1-induced cell migration, and the induction of downstream signaling by a constitutively active form of G(12 alpha). Furthermore, germinial center B lymphocytes, which are refractory to stromal-derived factor-1-triggered migration, express high levels of RGS1, These results indicate that RGS proteins can profoundly effect the directed migration of lymphoid cells. C1 NIAID, Immunoregulat Lab, B Cell Mol Immunol Sect, NIH, Bethesda, MD 20892 USA. NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. Univ Texas, SW Med Ctr, Dept Pharmacol, Dallas, TX 75235 USA. RP Kehrl, JH (reprint author), NIAID, Immunoregulat Lab, B Cell Mol Immunol Sect, NIH, Bldg 10,Room 11B-13,10 Ctr Dr,MSC 1876, Bethesda, MD 20892 USA. NR 58 TC 75 Z9 75 U1 0 U2 2 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD FEB 15 PY 2000 VL 164 IS 4 BP 1829 EP 1838 PG 10 WC Immunology SC Immunology GA 283VK UT WOS:000085296600028 PM 10657631 ER PT J AU Zappacosta, F Tabaczewski, P Parker, KC Coligan, JE Stroynowski, I AF Zappacosta, F Tabaczewski, P Parker, KC Coligan, JE Stroynowski, I TI The murine liver-specific nonclassical MHC class I molecule Q10 binds a classical peptide repertoire SO JOURNAL OF IMMUNOLOGY LA English DT Article ID MAJOR HISTOCOMPATIBILITY COMPLEX; TISSUE-SPECIFIC EXPRESSION; CELL-MEDIATED LYSIS; ANTIGEN PRESENTATION; T-CELLS; HLA-E; SOLUBLE FORM; QA-2 ANTIGEN; MOUSE; GENE AB The biological properties of the nonclassical class I MHC molecules secreted into blood and tissue fluids are not currently understood. To address this issue, we studied the murine Q10 molecule, one of the most abundant, soluble class Ib molecules. Mass spectrometry analyses of hybrid Q10 polypeptides revealed that alpha 1 alpha 2 domains of Q10 associate with 8-9 long peptides similar to the classical class I MHC ligands, Several of the sequenced peptides matched intracellularly synthesized murine proteins, This finding and the observation that the Q10 hybrid assembly is TAP2-dependent supports the notion that Q10 groove is loaded by the classical class I Ag presentation pathway. Peptides eluted from Q10 displayed a binding motif typical of H-2K, D, and L ligands, They carried conserved residues at P2 (Gly), P6 (Leu), and P omega (Phe/Leu), The role of these residues as anchors/auxiliary anchors was confirmed by Ala substitution experiments. The Q10 peptide repertoire was heterogeneous, with 75% of the groove occupied by a multitude of diverse peptides; however, 25% of the molecules bound a single peptide identical to a region of a TCR V beta-chain. Since this peptide did not display enhanced binding affinity for Q10 nor does its origin and sequence suggest that it is functionally significant, we propose that the nonclassical class I groove of Q10 resembles H-2K, D, and L grooves more than the highly specialized clefts of nonclassical class I Ags such as Qa-1, HLA-E, and M3. C1 NIAID, Lab Allerg Dis, NIH, Rockville, MD 20852 USA. Univ Texas, SW Med Ctr, Ctr Immunol, Dallas, TX 75235 USA. Univ Texas, SW Med Ctr, Dept Microbiol, Dallas, TX 75235 USA. Univ Texas, SW Med Ctr, Dept Internal Med, Dallas, TX 75235 USA. RP Stroynowski, I (reprint author), NIAID, Lab Allerg Dis, NIH, Twinbrook 2,Room 205, Rockville, MD 20852 USA. OI Parker, Kenneth/0000-0002-6282-2478 FU NIAID NIH HHS [AI19624, AI37818] NR 67 TC 15 Z9 15 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 FEB 15 PY 2000 VL 164 IS 4 BP 1906 EP 1915 PG 10 WC Immunology SC Immunology GA 283VK UT WOS:000085296600037 PM 10657640 ER PT J AU Weinberg, AD Rivera, MM Prell, R Morris, A Ramstad, T Vetto, JT Urba, WJ Alvord, G Bunce, C Shields, J AF Weinberg, AD Rivera, MM Prell, R Morris, A Ramstad, T Vetto, JT Urba, WJ Alvord, G Bunce, C Shields, J TI Engagement of the OX-40 receptor in vivo enhances antitumor immunity SO JOURNAL OF IMMUNOLOGY LA English DT Article ID T-CELL ACTIVATION; COLONY-STIMULATING FACTOR; OX40 LIGAND; IN-VIVO; DENDRITIC CELLS; TUMOR-CELLS; AUTOIMMUNE ENCEPHALOMYELITIS; B7 COSTIMULATION; LYMPH-NODES; SPINAL-CORD AB The OX-40 receptor (OX-40R), a member of the TNFR family, is primarily expressed on activated CD4(+) T lymphocytes. Engagement of the OX-40R, with either OX-40 ligand (OX-40L) or an Ab agonist, delivers a strong costimulatory signal to effector T cells. OX-40R(+) T cells isolated from inflammatory lesions in the CNS of animals with experimental autoimmune encephalomyelitis are the cells that respond to autoantigen (myelin basic protein) in vivo. We identified OX-40R(+) T cells within primary tumors and tumor-invaded lymph nodes of patients with cancer and hypothesized that they are the tumor-Ag-specific T cells. Therefore, we investigated whether engagement of the OX-40R in vivo during tumor priming would enhance a tumor-specific T cell response. Injection of OX-40L:Ig or anti-OX-40R in vivo during tumor priming resulted in a significant improvement in the percentage of tumor-free survivors (20-55%) in four different murine tumors derived from four separate tissues. This anti-OX-40R effect was dose dependent and accentuated tumor-specific T cell memory. The data suggest that engagement of the OX-40R in vivo augments tumor-specific priming by stimulating/expanding the natural repertoire of the host's tumor-specific CD4(+) T cells. The identification of OX-40R(+) T cells clustered around human tumor cells in vivo suggests that engagement of the OX-40R may be a practical approach for expanding tumor-reactive T cells and thereby a method to improve tumor immunotherapy in patients with cancer. C1 Robert W Franz Canc Res Ctr, Earle A Chiles Res Inst, Prov Portland Med Ctr, Portland, OR 97213 USA. Oregon Hlth Sci Univ, Dept Surg, Sect Surg Oncol, Portland, OR 97201 USA. Cantab Pharmaceut, Cambridge, England. NCI, Comp Serv, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. NCI, Stat Serv, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RP Weinberg, AD (reprint author), Robert W Franz Canc Res Ctr, Earle A Chiles Res Inst, Prov Portland Med Ctr, 4805 NE Glisan, Portland, OR 97213 USA. FU NCI NIH HHS [1-RO1-CA81383-01] NR 55 TC 214 Z9 225 U1 0 U2 3 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD FEB 15 PY 2000 VL 164 IS 4 BP 2160 EP 2169 PG 10 WC Immunology SC Immunology GA 283VK UT WOS:000085296600067 PM 10657670 ER PT J AU Mattson, MP Zhu, HY Yu, J Kindy, MS AF Mattson, MP Zhu, HY Yu, J Kindy, MS TI Presenilin-1 mutation increases neuronal vulnerability to focal ischemia in vivo and to hypoxia and glucose deprivation in cell culture: Involvement of perturbed calcium homeostasis SO JOURNAL OF NEUROSCIENCE LA English DT Article DE Alzheimer's disease; dantrolene; endoplasmic reticulum; knock-in; stroke; transgenic ID AMYLOID-PRECURSOR PROTEIN; FAMILIAL ALZHEIMERS-DISEASE; HUMAN CORTICAL-NEURONS; SECRETED FORMS; BRAIN INJURY; BETA-PEPTIDE; MITOCHONDRIAL DYSFUNCTION; ENDOPLASMIC-RETICULUM; MUTANT PRESENILIN-1; HIPPOCAMPAL-NEURONS AB Many cases of early-onset inherited Alzheimer's disease (AD) are caused by mutations in the presenilin-1 (PS1) gene. Studies of cultured neural cells suggest that PS1 mutations result in perturbed cellular calcium homeostasis and may thereby render neurons vulnerable to apoptosis. In light of evidence that metabolic impairment plays a role in AD, that cerebral ischemia may be a risk factor for AD, and that individuals with AD have increased morbidity and mortality after stroke, we examined the impact of a PS1 mutation on neuronal vulnerability to ischemic injury. We report that the extent of brain injury after focal cerebral ischemia reperfusion is increased, and behavioral outcome is worsened, in PS1 mutant knock-in mice compared to wild-type mice. Cultured cortical neurons from PS1 mutant mice exhibit increased vulnerability to glucose deprivation and chemical hypoxia compared to their wild-type counterparts. Calcium imaging studies demonstrated enhanced elevation of intracellular calcium levels after glucose deprivation and chemical hypoxia in neurons from PS1 mutant mice. Agents that block calcium release from IP3- and ryanodine-sensitive stores (xestospongin and dantrolene, respectively) protected against the endangering action of the PS1 mutation. Our data suggest that presenilin mutations may promote neuronal degeneration in AD by increasing the sensitivity of neurons to age-related ischemia-like conditions. The data further suggest that drugs that stabilize endoplasmic reticulum calcium homeostasis may prove effective in suppressing the neurodegenerative process in AD patients. C1 NIA, Neurosci Lab, Baltimore, MD 21224 USA. Univ Kentucky, Sanders Brown Ctr Aging, Lexington, KY 40536 USA. Univ Kentucky, Dept Anat & Neurobiol, Lexington, KY 40536 USA. Univ Kentucky, Dept Biochem, Lexington, KY 40536 USA. Vet Affairs Med Ctr, Lexington, KY 40506 USA. RP Mattson, MP (reprint author), NIA, Neurosci Lab, GRC 4F01,5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012 NR 52 TC 136 Z9 143 U1 0 U2 3 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD FEB 15 PY 2000 VL 20 IS 4 BP 1358 EP 1364 PG 7 WC Neurosciences SC Neurosciences & Neurology GA 281VJ UT WOS:000085181200010 PM 10662826 ER PT J AU Asmus, SE Parsons, S Landis, SC AF Asmus, SE Parsons, S Landis, SC TI Developmental changes in the transmitter properties of sympathetic neurons that innervate the periosteum SO JOURNAL OF NEUROSCIENCE LA English DT Article DE cholinergic differentiation factor; neural crest; sweat glands; synapse formation; neuropoietic cytokines; transmitter plasticity; autonomic neurons ID VESICULAR ACETYLCHOLINE TRANSPORTER; VASOACTIVE-INTESTINAL-PEPTIDE; LEUKEMIA INHIBITORY FACTOR; RAT SWEAT GLANDS; NEUROTRANSMITTER PLASTICITY INVIVO; GENE-RELATED PEPTIDE; CHOLINE-ACETYLTRANSFERASE; DIFFERENTIATION FACTOR; NERVOUS-SYSTEM; SURGICAL SYMPATHECTOMY AB During the development of sweat gland innervation, interactions with the target tissue induce a change from noradrenergic to cholinergic and peptidergic properties. To determine whether the change in neurotransmitter properties that occurs in the sweat gland innervation occurs more generally in sympathetic neurons, we identified a new target of cholinergic sympathetic neurons in rat, the periosteum, which is the connective tissue covering of bone, and characterized the development of periosteal innervation of the sternum. During development, sympathetic axons grow from thoracic sympathetic ganglia along rib periosteum to reach the sternum. All sympathetic axons displayed catecholaminergic properties when they reached the sternum, but these properties subsequently disappeared. Many axons lacked detectable immunoreactivities for vesicular acetylcholine transporter and vasoactive intestinal peptide when they reached the sternum and acquired them after arrival. To determine whether periosteum could direct changes in the neurotransmitter properties of sympathetic neurons that innervate it, we transplanted periosteum to the hairy skin, a noradrenergic sympathetic target. We found that the sympathetic innervation of the transplant underwent a noradrenergic to cholinergic and peptidergic change. These results suggest that periosteum, in addition to sweat glands, regulates the neurotransmitter properties of the sympathetic neurons that innervate it. C1 NINDS, NIH, Bethesda, MD 20892 USA. RP Landis, SC (reprint author), NINDS, NIH, Bethesda, MD 20892 USA. FU NINDS NIH HHS [NS023678, NS09709] NR 64 TC 71 Z9 72 U1 0 U2 1 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD FEB 15 PY 2000 VL 20 IS 4 BP 1495 EP 1504 PG 10 WC Neurosciences SC Neurosciences & Neurology GA 281VJ UT WOS:000085181200023 PM 10662839 ER PT J AU McGowan, JA Pottern, L AF McGowan, JA Pottern, L TI Commentary on the Women's Health Initiative SO MATURITAS LA English DT Editorial Material DE Women's Health Initiative; postmenopausal women; heart disease; cancer; osteoporosis ID PREVENTION; DISEASE AB The Women's Health Initiative (WHI), established by the National Institutes of Health in 1991, is a long-term national health study that focuses on strategies for preventing heart disease, breast and colorectal cancer and osteoporosis in postmenopausal women. These chronic diseases are the major causes of death, disability and frailty in older women of all races and socioeconomic backgrounds. The WHI a 15-year multi-million dollar endeavor, and one of the largest U.S. prevention studies of its kind. The study involves over 161 000 women aged 50-79, and is one of the most definitive, far reaching clinical trials of women's health ever undertaken in the U.S. The WHI Clinical Trial and Observational Study will attempt to address many of the inequities in women's health research and provide practical information to women and their physicians about hormone replacement therapy, dietary patterns and calcium/vitamin D supplements, and their effects on the prevention of heart disease, cancer and osteoporosis. Emerging information from the NIH Women's Health Initiative and other studies of women's health begun in the 1990's should be changing the landscape of options for older women in the years to come. Published by Elsevier Science Ireland Ltd. All rights reserved. C1 NIAMSD, NIAMS, NIH, Bethesda, MD 20892 USA. NHLBI, Bethesda, MD 20892 USA. RP McGowan, JA (reprint author), NIAMSD, NIAMS, NIH, Natcher Bldg,Room 5AS-43E, Bethesda, MD 20892 USA. NR 7 TC 11 Z9 13 U1 0 U2 1 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0378-5122 J9 MATURITAS JI Maturitas PD FEB 15 PY 2000 VL 34 IS 2 BP 109 EP 112 DI 10.1016/S0378-5122(99)00109-7 PG 4 WC Geriatrics & Gerontology; Obstetrics & Gynecology SC Geriatrics & Gerontology; Obstetrics & Gynecology GA 284ZZ UT WOS:000085363900002 PM 10714904 ER PT J AU Chen, JH Le, SY Maizel, JV AF Chen, JH Le, SY Maizel, JV TI Prediction of common secondary structures of RNAs: a genetic algorithm approach SO NUCLEIC ACIDS RESEARCH LA English DT Article ID TARGET SEQUENCE; RIBOSOMAL-RNA; MESSENGER-RNA; ALIGNMENT; PROTEIN; ELEMENT; RRE AB In this study we apply a genetic algorithm to a set of RNA sequences to find common RNA secondary structures. Our method is a three-step procedure. At the first stage of the procedure for each sequence, a genetic algorithm is used to optimize the structures in a population to a certain degree of stability. In this step, the free energy of a structure is the fitness criterion for the algorithm. Next, for each structure, we define a measure of structural conservation with respect to those in other sequences. We use this measure in a genetic algorithm to improve the structural similarity among sequences for the structures in the population of a sequence. Finally, we select those structures satisfying certain conditions of structural stability and similarity as predicted common structures for a set of RNA sequences. We have obtained satisfactory results from a set of tRNA, 5S rRNA, rev response elements(RRE) of HIV-1 and RRE of HIV-2/SIV, respectively. C1 NCI, Frederick Canc Res & Dev Ctr, SAIC, Adv Biomed Comp Ctr, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, Lab Expt & Computat Biol, NIH, Frederick, MD 21702 USA. RP Chen, JH (reprint author), NCI, Frederick Canc Res & Dev Ctr, SAIC, Adv Biomed Comp Ctr, Frederick, MD 21702 USA. FU NCI NIH HHS [N01-CO-56000] NR 22 TC 63 Z9 66 U1 0 U2 4 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0305-1048 J9 NUCLEIC ACIDS RES JI Nucleic Acids Res. PD FEB 15 PY 2000 VL 28 IS 4 BP 991 EP 999 DI 10.1093/nar/28.4.991 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 284ZN UT WOS:000085362900019 PM 10648793 ER PT J AU Hu, XL Margolis, HS Purcell, RH Ebert, J Robertson, BH AF Hu, XL Margolis, HS Purcell, RH Ebert, J Robertson, BH TI Identification of hepatitis B virus indigenous to chimpanzees SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID NUCLEOTIDE-SEQUENCE; SURFACE-ANTIGEN; GENOME; HEPADNAVIRUS; WOODCHUCK; SUBTYPES AB Hepatitis B viruses (HBV) and related viruses, classified in the Hepadnaviridae family, are found in a wide variety of mammals and birds. Although the chimpanzee has been the primary experimental model of HBV infection, this species has not been considered a natural host for the virus. Retrospective analysis of 13 predominantly wild-caught chimpanzees with chronic HBV infection identified a unique chimpanzee HBV strain in 11 animals. Nucleotide and derived amino acid analysis of the complete HBV genome and the gene coding for the hepatitis B surface antigen (S gene) identified sequence patterns that could be used to reliably identify chimpanzee HBV. This analysis indicated that chimpanzee HBV is distinct from known human HBV genotypes and is closely related to HBVs previously isolated from a chimpanzee, gibbons, gorillas, and orangutans. C1 Ctr Dis Control & Prevent, Hepatitis Branch A33, Div Viral & Rickettsial Dis, Natl Ctr Infect Dis, Atlanta, GA 30333 USA. NIAID, Hepatitis Viruses Sect, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. RP Robertson, BH (reprint author), Ctr Dis Control & Prevent, Hepatitis Branch A33, Div Viral & Rickettsial Dis, Natl Ctr Infect Dis, 1600 Clifton Rd NE, Atlanta, GA 30333 USA. NR 21 TC 73 Z9 76 U1 0 U2 0 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD FEB 15 PY 2000 VL 97 IS 4 BP 1661 EP 1664 DI 10.1073/pnas.97.4.1661 PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 285VU UT WOS:000085409600069 PM 10677515 ER PT J AU Sawitzke, JA Austin, S AF Sawitzke, JA Austin, S TI Suppression of chromosome segregation defects of Escherichia coli muk mutants by mutations in topoisomerase I SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID BACILLUS-SUBTILIS; DNA GYRASE; BIPOLAR LOCALIZATION; POLAR LOCALIZATION; REPLICATION ORIGIN; PLASMID-PARTITION; P1 PLASMID; PROTEIN; GENE; CONDENSATION AB Escherichia coli muk mutants are temperature-sensitive and produce anucleate cells. A spontaneously occurring mutation was found in a Delta mukB::kan mutant strain that suppressed the temperature-sensitive phenotype and mapped in or near topA. the gene that encodes topoisomerase I. Previously characterized topA mutations, topA10 and topA66, were found to be general suppressors of muk mutants: they suppressed temperature sensitivity and anucleate cell production of cells containing null or point mutations in mukB and null mutations in mokE or mukF. The suppression correlated with excess negative supercoiling by DNA gyrase, and the gyrase inhibitor, coumermycin. reversed it. Defects in topA allow 99% of cell division events in muk null mutants to proceed without chromosome loss or loss of cell viability. This observation imposes important limitations on models for Muk activity and is consistent with a role for MukBEF in chromosome folding and DNA condensation. C1 NCI, ABL Basic Res Program, Frederick Canc Res & Dev Ctr, Ft Detrick, MD 21702 USA. RP Austin, S (reprint author), NCI, ABL Basic Res Program, Frederick Canc Res & Dev Ctr, Ft Detrick, MD 21702 USA. FU NIGMS NIH HHS [F32 GM016971, 5 F32 GM16971] NR 46 TC 123 Z9 126 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 FEB 15 PY 2000 VL 97 IS 4 BP 1671 EP 1676 DI 10.1073/pnas.030528397 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 285VU UT WOS:000085409600071 PM 10660686 ER PT J AU Caturegli, P Hejazi, M Suzuki, K Dohan, O Carrasco, N Kohn, LD Rose, NR AF Caturegli, P Hejazi, M Suzuki, K Dohan, O Carrasco, N Kohn, LD Rose, NR TI Hypothyroidism in transgenic mice expressing IFN-gamma in the thyroid SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID EXPERIMENTAL AUTOIMMUNE-THYROIDITIS; CULTURED HUMAN THYROCYTES; CLASS-II MHC; INTERFERON-GAMMA; THYROTROPIN RECEPTOR; GENE-EXPRESSION; ECTOPIC EXPRESSION; THYROGLOBULIN; DISEASE; TRANSCRIPTION AB IFN-gamma has been implicated with contradictory results in the pathogenetic process of autoimmune (Hashimoto's) thyroiditis, the most common cause of hypothyroidism in adults. To test whether the local production of IFN-gamma can lead to thyroid dysfunction. we have generated transgenic mice that express constitutively IFN-gamma in the thyroid follicular cells. This expression resulted in severe hypothyroidism. with growth retardation and disruption of the thyroid architecture. The hypothyroidism derived from a profound inhibition of the expression of the sodium iodide symporter gene. Taken together, these results indicate a direct role of IFN-gamma in the thyroid dysfunction that occurs in autoimmune thyroiditis. C1 Johns Hopkins Univ, Sch Med, Dept Pathol, Baltimore, MD 21205 USA. NIDDKD, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. Yeshiva Univ Albert Einstein Coll Med, Dept Mol Pharmacol, Bronx, NY 10461 USA. RP Caturegli, P (reprint author), Johns Hopkins Med Inst, Dept Pathol, Ross Bldg,Room 648,720 Rutland Ave, Baltimore, MD 21205 USA. RI Dohan, Orsolya/B-3750-2012 NR 40 TC 80 Z9 82 U1 0 U2 0 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD FEB 15 PY 2000 VL 97 IS 4 BP 1719 EP 1724 DI 10.1073/pnas.020522597 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 285VU UT WOS:000085409600079 PM 10677524 ER PT J AU Smith, JD Craig, AG Kriek, N Hudson-Taylor, D Kyes, S Fagen, T Pinches, R Baruch, DI Newbold, CI Miller, LH AF Smith, JD Craig, AG Kriek, N Hudson-Taylor, D Kyes, S Fagen, T Pinches, R Baruch, DI Newbold, CI Miller, LH TI Identification of a Plasmodium falciparum intercellular adhesion molecule-1 binding domain: A parasite adhesion trait implicated in cerebral malaria SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID ERYTHROCYTE-MEMBRANE PROTEIN-1; INFECTED ERYTHROCYTES; ANTIGENIC VARIATION; RECEPTOR; ADHERENCE; CD36; SEQUESTRATION; FAMILY; LIGAND; PFEMP1 AB Binding of infected erythrocytes to brain venules is a central pathogenic event in the lethal malaria disease complication, cerebral malaria. The only parasite adhesion trait linked to cerebral sequestration is binding to intercellular adhesion molecule-1 (ICAM-1). In this report, we show that Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) binds ICAM-1, We have cloned and expressed PfEMP1 recombinant proteins from the A4tres parasite. Using heterologous expression in mammalian cells, the minimal ICAM-1 binding domain was a complex domain consisting of the second Duffy binding-like (DBL) domain and the C2 domain. Constructs that contained either domain alone did not bind ICAM-1. Based on phylogenetic criteria, there are five distinct PfEMP1 DBL types designated alpha, beta, gamma, delta, and epsilon. The DBL domain from the A4tres that binds ICAM-1 is DBL beta type. A PfEMP1 cloned from a distinct ICAM-1 binding variant, the A4 parasite, contains a DBL beta domain and a C2 domain in tandem arrangement similar to the A4tres PfEMP1, Anti-PfEMP1 antisera implicate the DBL beta domain from A4var PfEMP1 in ICAM-1 adhesion. The identification of a P. falciparum ICAM-1 binding domain may clarify mechanisms responsible for the pathogenesis of cerebral malaria and lead to interventions or vaccines that reduce malarial disease. C1 NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. John Radcliffe Hosp, Nuffield Dept Med, Inst Mol Med, Mol Parasitol Grp, Oxford OX3 9DU, England. RP Newbold, CI (reprint author), NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. OI Newbold, Chris/0000-0002-9274-3789 FU NIDDK NIH HHS [DK02700-37] NR 34 TC 155 Z9 158 U1 0 U2 7 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 FEB 15 PY 2000 VL 97 IS 4 BP 1766 EP 1771 DI 10.1073/pnas.040545897 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 285VU UT WOS:000085409600087 PM 10677532 ER PT J AU Pourquier, P Takebayashi, Y Urasaki, Y Gioffre, C Kohlhagen, G Pommier, Y AF Pourquier, P Takebayashi, Y Urasaki, Y Gioffre, C Kohlhagen, G Pommier, Y TI Induction of topoisomerase I cleavage complexes by 1-beta-D-arabinofuranosylcytosine (ara-C) in vitro and in ara-C-treated cells SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE camptothecin; DNA repair; DNA damage; nucleoside analog ID EUKARYOTIC TOPOISOMERASE; CYTO-TOXICITY; DNA-SYNTHESIS; CAMPTOTHECIN; MECHANISM; ARABINOSYLCYTOSINE; INHIBITION; TOPOTECAN; COVALENT; ENZYME AB 1-beta-D-Arabinofuranosylcytosine (Ara-C) is a nucleoside analog commonly used in the treatment of leukemias. Ara-C inhibits DNA polymerases and can be incorporated into DNA, Its mechanism of cytotoxicity is not fully understood. Using oligonucleotides and purified human topoisomerase I (top1), we found a 4- to 6-fold enhancement of top1 cleavage complexes when ara-e was incorporated at the +1 position (immediately 3') relative to a unique top1 cleavage site. This enhancement was primarily due to a reversible inhibition of top1-mediated DNA religation. Because ara-C incorporation is known to alter base stacking and sugar puckering at the misincorporation site and at the neighboring base pairs, the observed inhibition of religation at the ara-C site suggests the importance of the alignment of the 5'-hydroxyl end for religation with the phosphate group of the top1 phosphotyrosine bond. This study also demonstrates that ara-C treatment and DNA incorporation trap top1 cleavage complexes in human leukemia cells. Finally, we report that camptothecin-resistant mouse P388/ CPT45 cells with no detectable top1 are crossresistant to ara-C, which suggests that top1 poisoning is a potential mechanism for ara-C cytotoxicity. C1 NCI, Mol Pharmacol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Pommier, Y (reprint author), NCI, Mol Pharmacol Lab, Div Basic Sci, NIH, Bldg 37,Room 5D02, Bethesda, MD 20892 USA. NR 42 TC 81 Z9 83 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 FEB 15 PY 2000 VL 97 IS 4 BP 1885 EP 1890 DI 10.1073/pnas.97.4.1885 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 285VU UT WOS:000085409600108 PM 10677551 ER PT J AU Kim, HJ Fay, MP Feuer, EJ Midthune, DN AF Kim, HJ Fay, MP Feuer, EJ Midthune, DN TI Permutation tests for joinpoint regression with applications to cancer rates SO STATISTICS IN MEDICINE LA English DT Article ID PROSTATE-CANCER AB The identification of changes in the recent trend is an important issue in the analysis of cancer mortality and incidence data. We apply a joinpoint regression model to describe such continuous changes and use the grid-search method to fit the regression function with unknown joinpoints assuming constant variance and uncorrelated errors. We find the number of significant joinpoints by performing several permutation tests, each of which has a correct significance level asymptotically. Each p-value is found using Monte Carlo methods, and the overall asymptotic significance level is maintained through a Bonferroni correction. These tests are extended to the situation with non-constant variance to handle rates with Poisson variation and possibly autocorrelated errors. The performance of these tests are studied via simulations and the tests are applied to U.S. prostate cancer incidence and mortality rates. Copyright (C) 2000 John Wiley & Sons, Ltd. C1 Syracuse Univ, Dept Math, Syracuse, NY 13244 USA. NCI, Bethesda, MD 20892 USA. RP Kim, HJ (reprint author), Syracuse Univ, Dept Math, 215 Carnegie Bldg, Syracuse, NY 13244 USA. RI Fay, Michael/A-2974-2008; OI Fay, Michael P./0000-0002-8643-9625 NR 27 TC 1275 Z9 1330 U1 4 U2 58 PU JOHN WILEY & SONS LTD PI W SUSSEX PA BAFFINS LANE CHICHESTER, W SUSSEX PO19 1UD, ENGLAND SN 0277-6715 J9 STAT MED JI Stat. Med. PD FEB 15 PY 2000 VL 19 IS 3 BP 335 EP 351 DI 10.1002/(SICI)1097-0258(20000215)19:3<335::AID-SIM336>3.3.CO;2-Q PG 17 WC Mathematical & Computational Biology; Public, Environmental & Occupational Health; Medical Informatics; Medicine, Research & Experimental; Statistics & Probability SC Mathematical & Computational Biology; Public, Environmental & Occupational Health; Medical Informatics; Research & Experimental Medicine; Mathematics GA 278NZ UT WOS:000084997000005 PM 10649300 ER PT J AU Giordano, V Jin, DY Rekosh, D Jeang, KT AF Giordano, V Jin, DY Rekosh, D Jeang, KT TI Intravirion targeting of a functional anti-human immunodeficiency virus ribozyme directed to pol SO VIROLOGY LA English DT Article ID GENE-THERAPY; REGULATED EXPRESSION; RAS RIBOZYME; RNA ENZYMES; STEM-CELLS; IN-VIVO; TYPE-1; REPLICATION; INFECTION; HIV-1 AB Ribozymes are catalytic RNAs that offer several advantages as specific therapeutic genes against human immunodeficiency virus type 1 (HIV-1). Significant challenges in antiviral uses of ribozymes include (1) how best to express and to deliver this agent and (2) what is the best locale to target ribozymes against HIV-1 RNA. To explore the former, we have previously characterized several vector systems for efficient expression/delivery of anti-HIV-1 ribozymes (Dropulic et al, 1992; Dropulic and Jeang, 1994a; Smith et al. 1997). Here, to investigate an optimal locale for ribozyme-targeting, we asked whether it might be advantageous to direct ribozymes into HIV-1 virions as opposed to the more conventional approach of targeting ribozymes into infected cells. Two series of experiments were performed. First, we demonstrated that anti-HIV-1 ribozymes could indeed be packaged specifically and efficiently into virions. Second, we compared the virus suppressing activity of a packageable ribozyme with its counterpart, which cannot be packaged into HIV-1 virions. Our results showed that although both ribozymes cleaved HIV-1 genomic RNA in vitro with equivalent efficiencies, the former ribozyme demonstrated significantly higher virus-suppressing activity than the latter. These findings provide proof-of-principle that to combat productive HIV-1 replication, intravirion targeting is more effective than intracellular targeting of ribozymes. (C) 2000 Academic Press. C1 NIAID, Mol Virol Sect, Mol Microbiol Lab, Bethesda, MD 20892 USA. Univ Virginia, Dept Microbiol, Charlottesville, VA 22908 USA. Univ Virginia, Myles H Thaler Ctr AIDS & Human Retrovirus Res, Charlottesville, VA 22908 USA. RP Jeang, KT (reprint author), NIAID, Mol Virol Sect, Mol Microbiol Lab, Bldg 4,Room 302,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Jeang, Kuan-Teh/A-2424-2008 NR 43 TC 13 Z9 15 U1 0 U2 1 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0042-6822 J9 VIROLOGY JI Virology PD FEB 15 PY 2000 VL 267 IS 2 BP 174 EP 184 DI 10.1006/viro.1999.0112 PG 11 WC Virology SC Virology GA 286FY UT WOS:000085434000005 PM 10662613 ER PT J AU Ho, NC Francomano, CA van Allen, M AF Ho, NC Francomano, CA van Allen, M TI Jeune asphyxiating thoracic dystrophy and short-rib polydactyly type III (Verma-Naumoff) are variants of the same disorder SO AMERICAN JOURNAL OF MEDICAL GENETICS LA English DT Article DE Jeune syndrome; short-rib polydactyly type III; intrafamilial variability; modifying loci AB Jeune syndrome (JS) and short-rib polydactyly syndrome type III (SRP type III) are autosomal recessive disorders characterized by short ribs and polydactyly. They are distinguished from each other by the more severe radiological and histological bone findings as well as the occurrence of facial anomalies, ambiguous genitalia, and occasionally, cloacal abnormalities in SRP type III, We present a family in which two children have mild JS and one has SRP type III as evidence that JS and SRP type III are variants of the same disorder. The intrafamilial variability may reflect the effects of modifying loci on gene expression, (C) 2000 Wiley-Liss, Inc. C1 NHGRI, Med Genet Branch, NIH, Bethesda, MD 20892 USA. Johns Hopkins Univ, Dept Med Genet, Baltimore, MD 21218 USA. British Columbia Childrens Hosp, Dept Med Genet, Vancouver, BC, Canada. RP Ho, NC (reprint author), NHGRI, Med Genet Branch, NIH, Bethesda, MD 20892 USA. NR 20 TC 31 Z9 33 U1 1 U2 3 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0148-7299 J9 AM J MED GENET JI Am. J. Med. Genet. PD FEB 14 PY 2000 VL 90 IS 4 BP 310 EP 314 DI 10.1002/(SICI)1096-8628(20000214)90:4<310::AID-AJMG9>3.0.CO;2-N PG 5 WC Genetics & Heredity SC Genetics & Heredity GA 281NB UT WOS:000085166200009 PM 10710229 ER PT J AU Nanni, L Croen, LA Lammer, EJ Muenke, M AF Nanni, L Croen, LA Lammer, EJ Muenke, M TI Holoprosencephaly: Molecular study of a California population SO AMERICAN JOURNAL OF MEDICAL GENETICS LA English DT Article DE holoprosencephaly; population-based study; Sonic ID MURINE HOMEOBOX GENE; SONIC-HEDGEHOG; RETINOIC ACID; MUTATIONS; MOUSE; EXPRESSION; INDUCTION; PROTEIN; HOMOLOG; EMBRYOS AB Holoprosencephaly (HPE) is a common developmental anomaly of the forebrain and midface in which the cerebral hemispheres fail to separate into distinct left and right halves. HPE is extremely heterogeneous. In addition to teratogenic agents, several genes are implicated in the cause of HPE, Using samples from a population-based birth defects registry in California, we performed a mutational analysis of the known HPE genes Sonic Hedgehog (SHH), ZIC2, and SIX3, in addition to two HPE candidate genes, TG-interacting factor (TGIF), and Patched (PTC), on a group of sporadic HPE patients. This is the first molecular study of HPE in a population-based sample of patients, Among these patients, a deletion in the homeodomain of SIX3 and several polymorphisms in SIX3 and TGIF were identified, No sequence changes were detected in SHH, ZIC2, and PTC, Our results suggest that mutations in the currently recognized HPE genes may explain <5% of all sporadic HPE cases. Published 2000 Wiley-Liss, Inc. C1 Natl Human Genome Res Inst, Med Genet Branch, NIH, Bethesda, MD 20892 USA. Univ Penn, Childrens Hosp Philadelphia, Sch Med, Dept Pediat, Philadelphia, PA 19104 USA. Univ Penn, Childrens Hosp Philadelphia, Sch Med, Dept Genet, Philadelphia, PA 19104 USA. Calif Birth Defects Monitoring Program, March Dimes Birth Defects Fdn, Emeryville, CA USA. Childrens Hosp, Div Med Genet, Oakland, CA 94609 USA. RP Muenke, M (reprint author), Natl Human Genome Res Inst, Med Genet Branch, NIH, 10 Ctr Dr,MSC 1852,Bldg 10,10C101, Bethesda, MD 20892 USA. FU NICHD NIH HHS [HD29862, HD28732] NR 39 TC 32 Z9 34 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0148-7299 J9 AM J MED GENET JI Am. J. Med. Genet. PD FEB 14 PY 2000 VL 90 IS 4 BP 315 EP 319 DI 10.1002/(SICI)1096-8628(20000214)90:4<315::AID-AJMG10>3.0.CO;2-Y PG 5 WC Genetics & Heredity SC Genetics & Heredity GA 281NB UT WOS:000085166200010 PM 10710230 ER PT J AU Erexson, GL Tindall, KR AF Erexson, GL Tindall, KR TI Reduction of diepoxybutane-induced sister chromatid exchanges by glutathione peroxidase and erythrocytes in transgenic Big Blue (R) mouse and rat fibroblasts SO MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS LA English DT Article DE Big Blue (R); SCE; diepoxybutane; detoxification ID CULTURED HUMAN-LYMPHOCYTES; SPECIES-DIFFERENCES; INHALATION TOXICITY; BLOOD CULTURES; WHOLE-BLOOD; 1,3-BUTADIENE; CARCINOGENICITY; BUTADIENE; INDUCTION; MICE AB We have investigated the effect of glutathione peroxidase (GSH-Px) and mammalian erythrocytes (RBCs) on spontaneous and diepoxybutane (DEB)-induced sister chromatid exchange (SCE) in primary Big Blue(R) mouse (BBM1) and Big Blue(R) rat (BBR1) fibroblasts. DEB is the putative carcinogenic metabolite of 1,3-butadiene (BD) for which inhalation exposure yields a high rate of malignancies in mice but not in rats. ED is metabolized differently in mice and rats, producing much higher levels of DEB in mice than in rats, which may partly explain the different carcinogenic responses. However, other factors may contribute to the observed differences in the rodent carcinogenic response to ED. DEB is a highly reactive compound. Upon epoxide hydrolysis, DEB can covalently bind to DNA bases. Likewise, DEB generates reactive oxygen species that, in turn, can either damage DNA or produce H2O2. Reduced glutathione (GSH) is known to play a role in the metabolism and detoxification of DEB; and GSH is reduced by GSH-Px in the presence of H2O2. GSH-Px is a constitutive enzyme that is found at high concentrations in mammalian RBCs. Therefore, we were interested in examining the role of RBCs and GSH-Px on DEB-induced SCE in rat and mouse cells for detection of possible differences in the species response. Transgenic BBM1 and BBR1 fibroblasts were treated with either 0, 2 or 4 mu M DEB plus 0, 2 or 20 units of GSH-Px with and without 2 x 10(8) species-specific RBCs. DEB effectively induced SCEs in both rat and mouse cells. The relative induction of SCEs in both cell types was comparable. Both GSH-Px and RBCs alone and in combination were effective in significantly reducing DEB-induced SCEs in both mouse and rat fibroblasts, although there was more variability in the SCE response in rat cells. The present study suggests that GSH-Px may be important in the detoxification of DEB-induced DNA damage that results in the formation of SCEs. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NIEHS, Mol Mutagenesis Grp, Lab Environm Carcinogenesis & Mutagenesis, Res Triangle Pk, NC 27709 USA. N Carolina State Univ, Coll Vet Med, Dept Anat Physiol Sci & Radiol, Comparat Biomed Sci Program, Raleigh, NC 27606 USA. RP Tindall, KR (reprint author), NIEHS, Mol Mutagenesis Grp, Lab Environm Carcinogenesis & Mutagenesis, POB 12233, Res Triangle Pk, NC 27709 USA. NR 33 TC 12 Z9 14 U1 0 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 FEB 14 PY 2000 VL 447 IS 2 BP 267 EP 274 DI 10.1016/S0027-5107(99)00218-3 PG 8 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology SC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology GA 293LD UT WOS:000085852400014 PM 10751610 ER PT J AU Ciccodicola, A D'Esposito, M Esposito, T Gianfrancesco, F Migliaccio, C Miano, MG Matarazzo, MR Vacca, M Franze, A Cuccurese, M Cocchia, M Curci, A Terracciano, A Torino, A Cocchia, S Mercadante, G Pannone, E Archidiacono, N Rocchi, M Schlessinger, D D'Urso, M AF Ciccodicola, A D'Esposito, M Esposito, T Gianfrancesco, F Migliaccio, C Miano, MG Matarazzo, MR Vacca, M Franze, A Cuccurese, M Cocchia, M Curci, A Terracciano, A Torino, A Cocchia, S Mercadante, G Pannone, E Archidiacono, N Rocchi, M Schlessinger, D D'Urso, M TI Differentially regulated and evolved genes in the fully sequenced Xq/Yq pseudoautosomal region SO HUMAN MOLECULAR GENETICS LA English DT Article ID HUMAN SEX-CHROMOSOMES; HOMOLOGY REGION; X-CHROMOSOMES; INACTIVATION; EVOLUTION; DNA; IDENTIFICATION; ORGANIZATION; EXPRESSION; RESOLUTION AB Human sex chromosomes, which are morphologically and genetically different, share few regions of homology, Among them, only pseudoautosomal regions (PARs) pair and recombine during meiosis, To better address the complex biology of these regions, we sequenced the telomeric 400 kb of the long arm of the human X chromosome, including 330 kb of the human Xq/YqPAR and the telomere, Sequencing reveals subregions with distinctive regulatory and evolutionary features. The proximal 295 kb contains two genes inactivated on both the inactive X and Y chromosomes [SYBL1 and a novel homologue (HSPRy3) of Drosophila sprouty], The GC-rich distal 35 kb, added in stages and much later in evolution, contains the X/Y expressed gene IL9R and a novel gene, CXYorf1, only 5 kb from the Xq telomere, These properties make Xq/YqPAR a model for studies of region-specific gene inactivation, telomere evolution, and involvement in sex-limited conditions. C1 CNR, Int Inst Genet & Biophys, I-80125 Naples, Italy. Univ Bari, Genet Inst, I-70126 Bari, Italy. NIA, GRC, NIH, Baltimore, MD 21224 USA. RP D'Urso, M (reprint author), CNR, Int Inst Genet & Biophys, Via Marconi 10, I-80125 Naples, Italy. RI terracciano, antonio/B-1884-2008; Miano, Maria Giuseppina/E-9056-2011; Franze', Annamaria/H-2539-2012; Vacca, Marcella/B-9513-2015; OI Miano, Maria Giuseppina/0000-0003-1396-9673; Vacca, Marcella/0000-0002-5495-9183; Franze', Annamaria/0000-0002-4392-9739; archidiacono, nicoletta/0000-0002-1439-9521; Esposito, Teresa/0000-0002-7879-8441 FU Telethon [E.0869, E.0672, E.0927, E.0962] NR 43 TC 67 Z9 77 U1 2 U2 3 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0964-6906 J9 HUM MOL GENET JI Hum. Mol. Genet. PD FEB 12 PY 2000 VL 9 IS 3 BP 395 EP 401 DI 10.1093/hmg/9.3.395 PG 7 WC Biochemistry & Molecular Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Genetics & Heredity GA 285GF UT WOS:000085380800010 PM 10655549 ER PT J AU Carter, MG Johns, MA Zeng, XB Zhou, L Zink, MC Mankowski, JL Donovan, DM Baylin, SB AF Carter, MG Johns, MA Zeng, XB Zhou, L Zink, MC Mankowski, JL Donovan, DM Baylin, SB TI Mice deficient in the candidate tumor suppressor gene Hic1 exhibit developmental defects of structures affected in the Miller-Dieker syndrome SO HUMAN MOLECULAR GENETICS LA English DT Article ID PROTEIN INTERACTION MOTIF; HUMAN BREAST-CANCER; DNA HYPERMETHYLATION; DROSOPHILA METAMORPHOSIS; LUNG CANCERS; EXPRESSION; DELETION; 17P13.3; DOMAIN; P53 AB HIC1 is a candidate tumor suppressor gene which is frequently hypermethylated in human tumors, and its location within the Miller-Dieker syndrome's critical deletion region at chromosome 17p13.3 makes it a candidate gene for involvement in this gene deletion syndrome. To study the function of murine Hid in development, we have created Hic1-deficient mice. These animals die perinatally and exhibit varying combinations of gross developmental defects throughout the second half of development, including acrania, exencephaly, cleft palate, limb abnormalities and omphalocele. These findings demonstrate a role for Hid in the development of structures affected in the Miller-Dieker syndrome, and provide functional evidence to strengthen its candidacy as a gene involved in this disorder. C1 Johns Hopkins Med Inst, Ctr Oncol, Baltimore, MD 21231 USA. Johns Hopkins Univ, Sch Med, Grad Program Human Genet & Mol Biol, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Grad Program Cellular & Mol Med, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Div Comparat Med, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Dept Pathol, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Dept Mol Microbiol & Immunol, Baltimore, MD 21205 USA. NIA, Transgen & Knockout Facil, NIH, Baltimore, MD 21224 USA. RP Baylin, SB (reprint author), Johns Hopkins Med Inst, Ctr Oncol, Baltimore, MD 21231 USA. RI Carter, Mark/B-5089-2010 FU NCI NIH HHS [CA43318] NR 44 TC 86 Z9 89 U1 0 U2 2 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0964-6906 J9 HUM MOL GENET JI Hum. Mol. Genet. PD FEB 12 PY 2000 VL 9 IS 3 BP 413 EP 419 DI 10.1093/hmg/9.3.413 PG 7 WC Biochemistry & Molecular Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Genetics & Heredity GA 285GF UT WOS:000085380800012 PM 10655551 ER PT J AU Piscitelli, SC Burstein, AH Chaitt, D Alfaro, RM Falloon, J AF Piscitelli, SC Burstein, AH Chaitt, D Alfaro, RM Falloon, J TI Indinavir concentrations and St John's wort SO LANCET LA English DT Article AB St John's wort reduced the area under the curve of the HIV-1 protease inhibitor indinavir by a mean of 57% (SD 19) and decreased the extrapolated 8-h indinavir trough by 81% (16) in healthy volunteers. A reduction in indinavir exposure of this magnitude could lead to the development of drug resistance and treatment failure. C1 NIAID, Dept Pharm, NIH, Bethesda, MD 20892 USA. NIAID, Warren G Magnuson Clin Ctr, NIH, Bethesda, MD 20892 USA. NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. RP Piscitelli, SC (reprint author), NIAID, Dept Pharm, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. NR 5 TC 588 Z9 604 U1 1 U2 10 PU LANCET LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0140-6736 J9 LANCET JI Lancet PD FEB 12 PY 2000 VL 355 IS 9203 BP 547 EP 548 DI 10.1016/S0140-6736(99)05712-8 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 287CN UT WOS:000085487400015 PM 10683007 ER PT J AU Purcell, RH Emerson, SU AF Purcell, RH Emerson, SU TI Hepatitis E virus infection SO LANCET LA English DT Letter C1 NIAID, Hepatitis Viruses Sect, NIH, Bethesda, MD 20892 USA. NIAID, Mol Hepatitis Sect, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. RP Purcell, RH (reprint author), NIAID, Hepatitis Viruses Sect, NIH, Bethesda, MD 20892 USA. NR 5 TC 19 Z9 19 U1 0 U2 2 PU LANCET LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0140-6736 J9 LANCET JI Lancet PD FEB 12 PY 2000 VL 355 IS 9203 BP 578 EP 578 DI 10.1016/S0140-6736(05)73231-1 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 287CN UT WOS:000085487400054 PM 10683033 ER PT J AU Kamitani, H Kameda, H Kelavkar, UP Eling, TE AF Kamitani, H Kameda, H Kelavkar, UP Eling, TE TI A GATA binding site is involved in the regulation of 15-lipoxygenase-1 expression in human colorectal carcinoma cell line, Caco-2 SO FEBS LETTERS LA English DT Article DE 15-lipoxygenase-1; GATA; differentiation; transcription factor ID TRANSCRIPTION FACTOR GATA-1; ARACHIDONATE 12-LIPOXYGENASES; EPITHELIAL-CELLS; DEVELOPING HEART; SODIUM-BUTYRATE; DIFFERENTIATION; PROTEIN; GENES; INTERLEUKIN-13; INTESTINE AB The data presented implicate a GATA binding site in the transcriptional regulation of 15-lipoxygenase-1 (15-LO-1) gene expression in human colorectal carcinoma Caco-2 cells. High expression of GATA-6 mRNA and protein was observed, while GATA-4 mRNA was expressed at a very low level in Caco-2 cells, The expression of GATA-6 was down-regulated, while 15-LO-1 expression was dramatically up-regulated after treatment with sodium butyrate (NaBT), A study using an electrophoretic mobility shift assay indicated that a GATA binding site of the 15-LO-1 promoter region binds to GATA proteins present in both undifferentiated and, to a lesser extent, NaBT-treated (differentiated) Caco-2 cells. Moreover, that DNA binding shift band was disrupted after the addition of GATA-6 antibody in a supershift assay in the absence of NaBT, suggesting that GATA-6 is bound to the GATA binding site of the 15-LO-1 promoter in undifferentiated cells, In contrast, the addition of GATA-6 antibody did not affect the DNA binding ability in NaBT-induced differentiated cells. On the other hand, mutation of the GATA site of the 15-LO-1 promoter decreased the transactivation of the 15-LO-1 promoter as measured by luciferase activity in both FBS and NaBT cultured cells, indicating an unknown GATA binding protein to up-regulate 15-LO-1 expression. These implicate the GATA site at -240 of the proximal region of the 15-LO-1 promoter in the basic transcription of 15-LO-1 gene expression in Caco-2 cells, with GAT4-6 acting to repress 15-LO-1 expression, (C) 2000 Federation of European Biochemical Societies. C1 NIEHS, Mol Carcinogenesis Lab, Res Triangle Pk, NC 27709 USA. Emory Univ, Div Renal, Atlanta, GA 30322 USA. Emory Univ, Ctr Glomerulonephritis, Atlanta, GA 30322 USA. RP Eling, TE (reprint author), NIEHS, Mol Carcinogenesis Lab, POB 12233, Res Triangle Pk, NC 27709 USA. EM eling@niehs.nih.gov NR 50 TC 31 Z9 31 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 FEB 11 PY 2000 VL 467 IS 2-3 BP 341 EP 347 DI 10.1016/S0014-5793(00)01155-8 PG 7 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 285ZF UT WOS:000085417700042 PM 10675566 ER PT J AU Kaye, FJ Modi, S Ivanovska, I Koonin, EV Thress, K Kubo, A Kornbluth, S Rose, MD AF Kaye, FJ Modi, S Ivanovska, I Koonin, EV Thress, K Kubo, A Kornbluth, S Rose, MD TI A family of ubiquitin-like proteins binds the ATPase domain of Hsp-70-like Stch SO FEBS LETTERS LA English DT Article DE cell cycle; Hsp70; Stch; ubiquitin-like gene ID HEAT-SHOCK PROTEINS; DNA-REPAIR; MOLECULAR CHAPERONE; UBA DOMAIN; IN-VITRO; ENCODES; BAG-1; ALIGNMENT; PATHWAY; HSC70 AB We have isolated two human ubiquitin-like (UbL) proteins that bind to a short peptide within the ATPase domain of the Hsp70-like Stch protein. Chap1 is a duplicated homologue of the yeast Dsk2 gene that is required for transit through the G2/M phase of the cell cycle and expression of the human full-length cDNA restored viability and suppressed the G2/M arrest phenotype of dsk2 Delta rad23 Delta Saccharomyces cerevisiae mutants. Chap2 is a homologue for Xenopus scythe which is an essential component of reaper-induced apoptosis in egg extracts, While the S-terminal UbL domains were not essential for Stch binding, Chap1/Dsk2 contains a Sti1-like repeat sequence that is required far binding to Stch and is also conserved in the Hsp70 binding proteins, Hip and p60/Sti1/Hop. These findings extend the association between Hsp70 members and genes encoding UbL sequences and suggest a broader role for the Hsp70-like ATPase family in regulating cell cycle and cell death events, (C) 2000 Federation of European Biochemical Societies. C1 NCI, Med Branch, Div Clin Sci, Bethesda, MD 20889 USA. Natl Naval Med Ctr, Bethesda, MD 20889 USA. Princeton Univ, Dept Mol Biol, Princeton, NJ 08544 USA. NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20814 USA. Duke Univ, Med Ctr, Dept Pharmacol & Canc Biol, Durham, NC 27710 USA. RP Kaye, FJ (reprint author), USN Hosp, NCI, Bldg 8,Rm 5101, Bethesda, MD 20889 USA. RI kaye, frederic/E-2437-2011; OI Rose, Mark/0000-0003-1112-4765 FU NIGMS NIH HHS [R01 GM56518] NR 30 TC 63 Z9 68 U1 0 U2 3 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0014-5793 J9 FEBS LETT JI FEBS Lett. PD FEB 11 PY 2000 VL 467 IS 2-3 BP 348 EP 352 DI 10.1016/S0014-5793(00)01135-2 PG 5 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 285ZF UT WOS:000085417700043 PM 10675567 ER PT J AU Hall, SK Armstrong, DL AF Hall, SK Armstrong, DL TI Conditional and unconditional inhibition of calcium-activated potassium channels by reversible protein phosphorylation SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID CA2+-ACTIVATED K+ CHANNEL; THYROTROPIN-RELEASING-HORMONE; VASOACTIVE INTESTINAL PEPTIDE; KINASE-A; BETA-SUBUNIT; CYCLIC-AMP; DIFFERENTIAL MODULATION; HIPPOCAMPAL-NEURONS; LARGE-CONDUCTANCE; PHOSPHATASE 2A AB Large conductance, calcium-activated potassium channels (BKCa or maxi-K) are important determinants of membrane excitability in many cell types. We used patch clamp techniques to study the biochemical regulatiom of native BKCa channel proteins by endogenous Ser/Thr-directed protein kinases and phosphatases in cell-free membrane patches from rat pituitary tumor cells (GE(4)C(1)). When protein kinase activity was blocked by removing ATP, endogenous protein phosphatases slowly increased BKCa channel activity approximately 3-fold. Dephosphorylated channels could be activated fully by physiological increases in cytoplasmic calcium or membrane depolarization. In contrast, endogenous protein kinases inhibited BKCa channel activity at two functionally distinct sites. A closely associated, cAMP-dependent protein kinase rapidly reduced channel activity in a conditional manner that could be overcome completely by increasing cytoplasmic free calcium 3-fold or 20 mV further depolarization. Phosphorylation at a pharmacologically distinct site inhibited channel activity unconditionally by reducing availability to approximately half that of maximum at all physiological calcium and voltages. Conditional versus unconditional inhibition of BKCa channel activity through different protein kinases provides cells with a powerful computational mechanism for regulating membrane excitability. C1 Cardiff Univ, Cardiff Sch Biosci, Physiol Unit, Cardiff CF10 3US, S Glam, Wales. NIEHS, Lab Signal Transduct, NIH, Res Triangle Pk, NC 27709 USA. RP Hall, SK (reprint author), Cardiff Univ, Cardiff Sch Biosci, Physiol Unit, POB 911,Museum Ave, Cardiff CF10 3US, S Glam, Wales. NR 46 TC 43 Z9 43 U1 0 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD FEB 11 PY 2000 VL 275 IS 6 BP 3749 EP 3754 DI 10.1074/jbc.275.6.3749 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 283RJ UT WOS:000085288800006 PM 10660522 ER PT J AU Taylor, GA Hudson, E Resau, JH Vande Woude, GF AF Taylor, GA Hudson, E Resau, JH Vande Woude, GF TI Regulation of P311 expression by Met-hepatocyte growth factor/scatter factor and the ubiquitin/proteasome system SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID TUMORIGENICITY; PROTOONCOGENE; CELLS; PROTEASOME; METASTASIS; RNA AB P311 is a mouse cDNA originally identified for its high expression in late-stage embryonic brain and adult cer ebellum, hippocampus, and olfactory bulb. The protein product of P311, however, had not been identified previously, and its function remains unknown. We report here that P311 expression is regulated at multiple levels by pathways that control cellular transformation. P311 mRNA expression was decreased sharply in both neural and smooth muscle cells when the cells were transformed by coexpression of the oncogenic tyrosine kinase receptor Met and its ligand hepatocyte growth factor/scatter factor. The P311 mRNA was found to encode an 8-kDa polypeptide that was subject to rapid degradation by the lactacystin-sensitive ubiquitin/proteasome system and an unidentified metalloprotease, resulting in a protein half-life of about 5 min. These data suggest that P311 expression is dramatically decreased by several pathways that regulate cellular growth. C1 Adv Biosci Labs, Basic Res Program, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RP Vande Woude, GF (reprint author), Van Andel Res Inst, 201 Monroe Ave NW,Suite 400, Grand Rapids, MI 49503 USA. FU NCI NIH HHS [N01-CO-46000] NR 23 TC 22 Z9 30 U1 0 U2 1 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD FEB 11 PY 2000 VL 275 IS 6 BP 4215 EP 4219 DI 10.1074/jbc.275.6.4215 PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 283RJ UT WOS:000085288800070 PM 10660586 ER PT J AU Wang, F Chen, LF Arcucci, O Harvey, EV Bowers, B Xu, YH Hammer, JA Sellers, JR AF Wang, F Chen, LF Arcucci, O Harvey, EV Bowers, B Xu, YH Hammer, JA Sellers, JR TI Effect of ADP and ionic strength on the kinetic and motile properties of recombinant mouse myosin V SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID MELANOSOME DISTRIBUTION; UNCONVENTIONAL MYOSIN; SMOOTH-MUSCLE; MOTOR PROTEIN; LIGHT-CHAIN; GENE; MELANOCYTES; DOMAIN; ACTIN; TRANSPORT AB Mouse myosin V is a two-headed unconventional myosin with an extended neck that binds six calmodulins. Double-headed (heavy meromyosin-like) and single-headed (subfragment 1-like) fragments of mouse myosin V were expressed in Sf9 cells, and intact myosin V was purified from mouse brain. The actin-activated MgATPase of the tissue-purified myosin V, and its expressed fragments had a high V-max and a low K-ATPase. Calcium regulated the MgATPase of intact myosin V but not of the fragments, Both the MgATPase activity and the in vitro motility were remarkably insensitive to ionic strength. Myosin V and its fragments translocated actin at very:low myosin surface densities. ADP markedly inhibited the actin-activated MgATPase activity and the in vitro motility, ADP dissociated from myosin V subfragment 1 at a rate of about 11.5 s(-1) under conditions where the V-max was 3.3 s(-1), indicating that, although not totally rate-limiting, ADP dissociation was close to the Fate-limiting step. The high affinity for actin and the slow rate of ADP release helps the myosin head to re main attached to actin for a large fraction of each ATPase cycle and allows actin filaments to be moved by only a few myosin V molecules in vitro. C1 NHLBI, Mol Cardiol Lab, NIH, Bethesda, MD 20892 USA. NHLBI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. NHLBI, Core Microscopy Facil, NIH, Bethesda, MD 20892 USA. RP Sellers, JR (reprint author), NHLBI, Mol Cardiol Lab, NIH, 10 Ctr Dr,Bldg 10,Rm 8N202, Bethesda, MD 20892 USA. NR 34 TC 104 Z9 104 U1 1 U2 4 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD FEB 11 PY 2000 VL 275 IS 6 BP 4329 EP 4335 DI 10.1074/jbc.275.6.4329 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 283RJ UT WOS:000085288800086 PM 10660602 ER PT J AU Uren, A Reichsman, F Anest, V Taylor, WG Muraiso, K Bottaro, DP Cumberledge, S Rubin, JS AF Uren, A Reichsman, F Anest, V Taylor, WG Muraiso, K Bottaro, DP Cumberledge, S Rubin, JS TI Secreted frizzled-related protein-1 binds directly to wingless and is a biphasic modulator of Wnt signaling SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID BETA-CATENIN; SPEMANN ORGANIZER; XENOPUS EMBRYOS; DROSOPHILA; GENE; FAMILY; ANTAGONIST; RECEPTORS; APOPTOSIS; CANCER AB Secreted Frizzled-related protein-1 (sFRP-1) contains a cysteine-rich domain homologous to the putative Wnt-binding site of Frizzleds. To facilitate the biochemical and biological analysis of sFRP-1, we developed a mammalian recombinant expression system that yields similar to 3 mg of purified protein/liter of conditioned medium. Using this recombinant protein, we demonstrated that sFRP-1 and Wg (wingless) interact in enzyme-linked immunosorbent and co-precipitation assays. Surprisingly, a derivative lacking the cysteine-rich domain retained the ability to bind Wg, Cross-linking experiments performed with radioiodinated sFRP-1 provided definitive evidence that sFRP-1 and Wg bind directly to each other. Besides detecting a cross-linked complex consistent in size with 1:1 stoichiometry of sFRP-1 and Wg, we also observed a larger complex whose size suggested the presence of a second sFRP-1 molecule. The formation of both complexes was markedly enhanced by an optimal concentration of exogenous heparin, emphasizing the potential importance of heparan-sulfate proteoglycan in Wnt binding and signaling. sFRP-1 exerted a biphasic effect on Wg activity in an armadillo stabilization assay, increasing armadillo level at low concentrations but reducing it at higher concentrations. These results provide new insights about the Wnt binding and biological activity of sFRPs. C1 NCI, Cellular & Mol Biol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. Univ Massachusetts, Dept Biochem & Mol Biol, Amherst, MA 01002 USA. RP Rubin, JS (reprint author), NCI, Cellular & Mol Biol Lab, Div Basic Sci, NIH, Bldg 37,Rm 1E24,37 Convent Dr,MSC 4255, Bethesda, MD 20892 USA. RI Bottaro, Donald/F-8550-2010 OI Bottaro, Donald/0000-0002-5057-5334 NR 57 TC 251 Z9 259 U1 0 U2 11 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD FEB 11 PY 2000 VL 275 IS 6 BP 4374 EP 4382 DI 10.1074/jbc.275.6.4374 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 283RJ UT WOS:000085288800092 PM 10660608 ER PT J AU Prasad, R Dianov, GL Bohr, VA Wilson, SH AF Prasad, R Dianov, GL Bohr, VA Wilson, SH TI FEN1 stimulation of DNA polymerase beta mediates an excision step in mammalian long patch base excision repair SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID FLAP ENDONUCLEASE-1; PURIFIED PROTEINS; SUBSTRATE-BINDING; PHOSPHATE LYASE; SITE REPAIR; RECONSTITUTION; PATHWAY; 3'-EXO/ENDONUCLEASE; REQUIREMENT; MECHANISM AB In mammalian cells, single-base lesions, such as uracil and abasic sites, appear to be repaired by at least two base excision repair (BER) subpathways: "single-nucleotide BER" requiring DNA synthesis of just one nucleotide and "long patch BER" requiring multi-nucleotide DNA synthesis. In single-nucleotide BER, DNA polymerase beta (beta-pol) accounts for both gap filling DNA synthesis and removal of the 5'-deoxyribose phosphate (dRP) of the abasic site, whereas the involvement of various DNA polymerases in long patch BER is less well understood. Recently, we found that beta-pol plays a role in mammalian cell extract-mediated long patch BER, in that formation of a key excision product, 5'-dRP-trinucleotide (5'-dRP-N-3), is dependent upon beta-pol (Dianov, G. L., Prasad, R., Wilson, S. H., and Bohr, V.A. (1999) J, Biol, Chem. 274, 13741-13743). The structure-specific endonuclease flap endonuclease 1 (FEN1) has also been suggested to be involved in long patch BER excision, Here, we demonstrate by immunodepletion experiments that 5'-dRP-N-3 excision in long patch BER of uracil-DNA in a human lymphoid cell extract is, indeed, dependent upon FEN1, Next, we reconstituted the excision step of long patch BER using purified human proteins and an oligonucleotide substrate with 5'-dRP at the margin of a one-nucleotide gap. Formation of the excision product 5'-dRP-N-3 was dependent upon both strand displacement DNA synthesis by beta-pol and FEN1 excision, FEN1 stimulated strand displacement DNA synthesis of beta-pol, FEN1 acting either alone, or without DNA synthesis by beta-pol, produced a two-nucleotide excision product, 5'-dRP-N-1, but not 5'-dRP-N-3. These results demonstrate that human FEN1 and beta-pol can cooperate in long patch BER excision and specify the predominant excision product seen with a cell extract. C1 NIEHS, NIH, Struct Biol Lab, Res Triangle Pk, NC 27709 USA. NIA, NIH, Mol Genet Lab, Baltimore, MD 21224 USA. RP Wilson, SH (reprint author), NIEHS, NIH, Struct Biol Lab, 111 TW Alexander Dr,Bldg 101,Rm B246, Res Triangle Pk, NC 27709 USA. NR 40 TC 163 Z9 166 U1 0 U2 4 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD FEB 11 PY 2000 VL 275 IS 6 BP 4460 EP 4466 DI 10.1074/jbc.275.6.4460 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 283RJ UT WOS:000085288800103 PM 10660619 ER PT J AU Issaq, HJ Xu, HY Chan, KC Dean, MC AF Issaq, HJ Xu, HY Chan, KC Dean, MC TI Effect of temperature on the separation of DNA fragments by highperformance liquid chromatography and capillary electrophoresis: a comparative study SO JOURNAL OF CHROMATOGRAPHY B LA English DT Article DE temperature effects; DNA fragments ID POLYMERASE CHAIN-REACTION; INDUCED FLUORESCENCE DETECTION; REACTION PRODUCTS; GEL-ELECTROPHORESIS; RESOLUTION; OLIGONUCLEOTIDES; QUANTITATION; GRADIENTS AB This study investigates the effect of experimental temperature on the separation of DNA fragments, 21-587 bp, by both high-performance liquid chromatography (HPLC) and capillary electrophoresis (CE). The results show that the temperature plays an important role in the HPLC separation of DNA fragments. The optimum temperature was found to be between 40 and 50 degrees C for HPLC, while 25 degrees C was the optimum temperature for the CE separation. Also, although CE migration times became shorter, efficiency and resolution decreased with an increase in temperature from 25 to 50 degrees C, but the separation was not significantly affected, Also, the optimum HPLC temperature might be different depending on the fragment sizes to be resolved. (C) 2000 Elsevier Science B.V. All rights reserved. C1 NCI, SAIC Frederick, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RP Issaq, HJ (reprint author), NCI, SAIC Frederick, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. OI Dean, Michael/0000-0003-2234-0631 FU NCI NIH HHS [N01-CO-56000] NR 26 TC 10 Z9 12 U1 0 U2 3 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-4347 J9 J CHROMATOGR B JI J. Chromatogr. B PD FEB 11 PY 2000 VL 738 IS 2 BP 243 EP 248 DI 10.1016/S0378-4347(99)00544-7 PG 6 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 286ZU UT WOS:000085479400006 PM 10718642 ER PT J AU Bi, DQ Anderson, LW Shapiro, J Shapiro, A Grem, JL Takimoto, CH AF Bi, DQ Anderson, LW Shapiro, J Shapiro, A Grem, JL Takimoto, CH TI Measurement of plasma uracil using gas chromatography-mass spectrometry in normal individuals and in patients receiving inhibitors of dihydropyrimidine dehydrogenase SO JOURNAL OF CHROMATOGRAPHY B LA English DT Article DE uracil; eniluracil; dihydropyrimidine dehydrogenase ID ORAL 5-FLUOROURACIL PRODRUGS; BLOOD MONONUCLEAR-CELLS; 5-ETHYNYLURACIL 776C85; ANTITUMOR-ACTIVITY; ANTIVIRAL DRUG; SORIVUDINE; FLUOROURACIL; TEGAFUR; PHARMACOKINETICS; BIOAVAILABILITY AB A sensitive gas chromatographic-mass spectrometric method is described for reliably measuring endogenous uracil in 100 mu l of human plasma. Validation of this assay over a wide concentration range, 0.025 mu M to 250 mu M (0.0028 mu g/ml to 28 mu g/ml), allowed for the determination of plasma uracil in patients treated with agents such as eniluracil, an inhibitor of the pyrimidine catabolic enzyme, dihydropyrimidine dehydrogenase. Calibration standards were prepared in human plasma using the stable isotope, [N-15(2)]uracil, to avoid interference from endogenous uracil and 10 mu M 5-chlorouracil was added as the internal standard. Published by Elsevier Science B.V. C1 NCI, Dev Therapeut Dept, Med Branch, Div Clin Sci,Natl Naval Med Ctr, Bethesda, MD 20889 USA. US FDA, Ctr Drug Evaluat & Res, Lab Clin Pharmacol, Rockville, MD 20850 USA. RP Takimoto, CH (reprint author), NCI, Dev Therapeut Dept, Med Branch, Div Clin Sci,Natl Naval Med Ctr, Bldg 8,Room 5101, Bethesda, MD 20889 USA. NR 25 TC 24 Z9 28 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-4347 J9 J CHROMATOGR B JI J. Chromatogr. B PD FEB 11 PY 2000 VL 738 IS 2 BP 249 EP 258 DI 10.1016/S0378-4347(99)00528-9 PG 10 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 286ZU UT WOS:000085479400007 PM 10718643 ER PT J AU Maciejewski, MW Liu, DJ Prasad, R Wilson, SH Mullen, GP AF Maciejewski, MW Liu, DJ Prasad, R Wilson, SH Mullen, GP TI Backbone dynamics and refined solution structure of the N-terminal domain of DNA polymerase beta. Correlation with DNA binding and dRP lyase activity SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE DNA polymerase beta; N-15 relaxation; protein dynamics; conformational exchange; DNA binding ID N-15 NMR RELAXATION; BASE EXCISION-REPAIR; MODEL-FREE APPROACH; ROTATIONAL DIFFUSION ANISOTROPY; MAGNETIC-RESONANCE RELAXATION; SPECTRAL DENSITY-FUNCTIONS; CHEMICAL-SHIFT ANISOTROPY; COLI RIBONUCLEASE HI; SINGLE-STRANDED-DNA; ESCHERICHIA-COLI AB Mammalian DNA polymerase beta functions in the base excision DNA repair pathway filling in short patches (1-5 nt) in damaged DNA and removing deoxyribose 5'-phosphate from the 5'-side of damaged DNA. The backbone dynamics and the refined solution structure of the N-terminal domain of P-Pol have been characterized in order to establish the potential contribution(s) of backbone motion to the DNA binding and deoxyribose 5'-phosphate lyase function of this domain. The N-terminal domain is formed from four helices packed as two antiparallel pairs with a 60 degrees crossing between the pairs. The RMSD of the NMR conformers (residues 13-80) is 0.37 Angstrom for the backbone heavy atoms and 0.78 Angstrom for all heavy atoms. NMR characterization of the binding site(s) for a ssDNA-5mer, ssDNA-8mer, ssDNA-9mer, and dsDNA-12mer shows a consensus surface for the binding of these various DNA oligomers, that surrounds and includes the deoxyribose 5'-phosphate lyase active site region. Connection segments between helices 1 and 2 and between helices 3 and 4 each contribute to DNA binding. Helix-3-turn-helix-4 forms a helix-hairpin-helix motif. The highly conserved hairpin sequence (LPGVG) displays a significant degree of picosecond time-scale motion within the backbone, that is possibly important for DNA binding at the phosphodiester backbone. An Omega-loop connecting helices 1 and 2 and helix-2 itself display significant exchange contributions (R-ex) at the backbone amides due to apparent conformational type motion on a millisecond time-scale. This motion is likely important in allowing the Omega-loop and helix-2 to shift toward, and productively interact with, gapped DNA. The deoxyribose 5'-phosphate lyase catalytic residues that include K72 which forms the Schiff's base, Y39 which is postulated to promote proton transfer to the aldehyde, and K35 which assists in phosphate elimination, show highly restricted backbone motion. H34, which apparently participates in detection of the abasic site hole and assists in the opening of the hemiacetal, shows conformational exchange. (C) 2000 Academic Press. C1 Univ Connecticut, Ctr Hlth, Dept Biochem, Farmington, CT 06032 USA. NIEHS, Res Triangle Pk, NC 27709 USA. RP Mullen, GP (reprint author), Univ Connecticut, Ctr Hlth, Dept Biochem, 263 Farmington Ave, Farmington, CT 06032 USA. FU NIGMS NIH HHS [GM18956, GM52738] NR 71 TC 60 Z9 61 U1 0 U2 4 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-2836 J9 J MOL BIOL JI J. Mol. Biol. PD FEB 11 PY 2000 VL 296 IS 1 BP 229 EP 253 DI 10.1006/jmbi.1999.3455 PG 25 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 283VC UT WOS:000085295900017 PM 10656829 ER PT J AU Poulos, SG Borlongan, CV AF Poulos, SG Borlongan, CV TI Artificial lighting conditions and melatonin alter motor performance in adult rats SO NEUROSCIENCE LETTERS LA English DT Article DE nocturnal; light; biological clock; circadian rhythm; rotorod; motor coordination ID CIRCADIAN-RHYTHMS; ENTRAINMENT AB Entrained circadian rhythms may modulate many behavioral: activities of animals and humans. In the present study, we examined whether lighting conditions and melatonin treatment participate behaviorally in the entrainment of circadian rhythms in the rodent. In experiment one, Sprague-Dawley:rats were introduced to the Rotorod test apparatus at nighttime or daytime and either with the lights on (4 lux) or in the dark. During nighttime tests, the exposure of rats to dark or I ig ht condition did not a Iter mean rev./min or length of times spent on the Rotorod. Interestingly, du ri ng daytime tests, animals exposed to light condition displayed significantly reduced mean rev./min (7.95 +/- 1.68), as well as length of rime on the Rotorod (41.07 +/- 3.45 s) compared with their performance in the dark condition (mean rev./min, 11.16 +/- 1.52; length of time spent on the Rotorod, 66.94 +/- 6.15 s). In experiment two, treatment with melatonin (1.5 mg/kg, orally administered at 1 h prior to testing) in animals introduced to the daytime test with exposure to light condition, restored the rev./min (12.90 +/- 1.26) and the time spent on the Rotorod (63.21 +/- 2.73 s) to near normal levels. Thus, we demonstrated here that exposure of nocturnal animals to their preferred dark condition and treatment with melatonin could enhance motor coordination. Published by Elsevier Science Ireland Ltd. C1 NIDA, Intramural Res Program, NIH, Baltimore, MD 21224 USA. Univ S Florida, Coll Med, Dept Neurosurg, Tampa, FL 33612 USA. RP Borlongan, CV (reprint author), NIDA, Intramural Res Program, NIH, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. OI Borlongan, Cesar/0000-0002-2966-9782 NR 23 TC 11 Z9 12 U1 0 U2 0 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0304-3940 J9 NEUROSCI LETT JI Neurosci. Lett. PD FEB 11 PY 2000 VL 280 IS 1 BP 33 EP 36 DI 10.1016/S0304-3940(99)00997-0 PG 4 WC Neurosciences SC Neurosciences & Neurology GA 282ZE UT WOS:000085251000009 PM 10696805 ER PT J AU Servant, G Weiner, OD Herzmark, P Balla, T Sedat, JW Bourne, HR AF Servant, G Weiner, OD Herzmark, P Balla, T Sedat, JW Bourne, HR TI Polarization of chemoattractant receptor signaling during neutrophil chemotaxis SO SCIENCE LA English DT Article ID PROTEIN-KINASE-B; PLECKSTRIN HOMOLOGY DOMAINS; PHOSPHATIDYLINOSITOL 3-KINASE; ACTIN POLYMERIZATION; BINDING; ACTIVATION; CDC42; CELLS; PHOSPHOINOSITIDES; LEUKOCYTES AB Morphologic polarity is necessary for chemotaxis of mammalian cells. As a probe of intracellular signals responsible for this asymmetry, the pleckstrin homology domain of the AKT protein kinase (or protein kinase B), tagged with the green fluorescent protein (PHAKT-GFP), was expressed in neutrophils. Upon exposure of cells to chemoattractant, PHAKT-GFP is recruited selectively to membrane at the cell's leading edge, indicating an internal signaling gradient that is much steeper than that of the chemoattractant. Translocation of PHAKT-GFP is inhibited by toxin-B from Clostridium difficile, indicating that it requires activity of one or more Rho guanosine triphosphatases. C1 Univ Calif San Francisco, Dept Cellular & Mol Pharmacol, San Francisco, CA 94143 USA. Univ Calif San Francisco, Dept Biochem & Biophys, San Francisco, CA 94143 USA. NICHHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. RP Bourne, HR (reprint author), Univ Calif San Francisco, Dept Cellular & Mol Pharmacol, San Francisco, CA 94143 USA. RI Weiner, Orion/F-2576-2011; OI Weiner, Orion/0000-0002-1778-6543; Balla, Tamas/0000-0002-9077-3335 FU NCI NIH HHS [CA-54427]; NIGMS NIH HHS [GM-27800, GM-25101, R01 GM027800-27] NR 48 TC 625 Z9 634 U1 2 U2 31 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 USA SN 0036-8075 J9 SCIENCE JI Science PD FEB 11 PY 2000 VL 287 IS 5455 BP 1037 EP 1040 DI 10.1126/science.287.5455.1037 PG 4 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 282WU UT WOS:000085245400047 PM 10669415 ER PT J AU Chang, AHC Horton, D Kovac, P AF Chang, AHC Horton, D Kovac, P TI Syntheses of fluorinated ligands to probe binding of antigenic determinants of Vibrio cholerae O : 1, serotypes Inaba and Ogawa, to antibodies SO TETRAHEDRON-ASYMMETRY LA English DT Article ID METHYL ALPHA-GLYCOSIDE; CARBOHYDRATE BENZYLIDENE ACETALS; CRYSTAL-STRUCTURE; O-POLYSACCHARIDE; REPEATING UNIT; DERIVATIVES; ANALOGS; ACID; O/1 AB Derivatives of methyl alpha-glycosides of antigenic determinants of Vibrio cholerae O:1, serotypes Inaba and Ogawa, specifically Fluorinated at position 2' or 4' have been synthesized by coupling the appropriately fluorinated derivatives of 3-deoxy-L-glycero-tetronic acid with the methyl oc-glycosides of perosamine. The compound having the fluorine atom at position 2 was obtained by electrophilic addition of fluorine to the glycal derived from the parent antigenic determinant, serotypes Inaba, using Selectfluor(TM) as a fluorination reagent. (C) 2000 Elsevier Science Ltd. All rights reserved. C1 NIDDK, NIH, LMC, Bethesda, MD 20892 USA. American Univ, Dept Chem, Washington, DC 20016 USA. RP Kovac, P (reprint author), NIDDK, NIH, LMC, Bethesda, MD 20892 USA. NR 26 TC 10 Z9 10 U1 0 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0957-4166 J9 TETRAHEDRON-ASYMMETR JI Tetrahedron: Asymmetry PD FEB 11 PY 2000 VL 11 IS 2 BP 595 EP 606 DI 10.1016/S0957-4166(99)00505-4 PG 12 WC Chemistry, Inorganic & Nuclear; Chemistry, Organic; Chemistry, Physical SC Chemistry GA 297JT UT WOS:000086080000027 ER PT J AU Minton, AP AF Minton, AP TI Protein folding: Thickening the broth SO CURRENT BIOLOGY LA English DT Article ID MOLECULAR CHAPERONES; LYSOZYME AB Recent results support the notion that macromolecular 'crowding' enhances protein aggregation, at the expense of correct folding. The results can be rationalised in terms of kinetic competition between distinct processes, taking into account the relative influence of crowding on each process. C1 NIH, Bethesda, MD 20892 USA. RP Minton, AP (reprint author), NIH, Bldg 8,Room 226, Bethesda, MD 20892 USA. NR 15 TC 35 Z9 36 U1 0 U2 3 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0960-9822 J9 CURR BIOL JI Curr. Biol. PD FEB 10 PY 2000 VL 10 IS 3 BP R97 EP R99 DI 10.1016/S0960-9822(00)00301-8 PG 3 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 284VT UT WOS:000085354100005 PM 10679316 ER PT J AU Cassman, M Hunter, T Pawson, T AF Cassman, M Hunter, T Pawson, T TI Proteins suggest form of their own database SO NATURE LA English DT Letter C1 Natl Inst Gen Med Sci, NIH, Bethesda, MD 20892 USA. Salk Inst, La Jolla, CA 92037 USA. Mt Sinai Hosp, Samuel Lunenfeld Res Inst, Toronto, ON M5G 1X5, Canada. RP Cassman, M (reprint author), Natl Inst Gen Med Sci, NIH, Bldg 45,2AN12B, Bethesda, MD 20892 USA. RI Pawson, Tony/E-4578-2013 NR 4 TC 3 Z9 3 U1 0 U2 0 PU MACMILLAN MAGAZINES LTD PI LONDON PA PORTERS SOUTH, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD FEB 10 PY 2000 VL 403 IS 6770 BP 591 EP 592 DI 10.1038/35001230 PG 2 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 283RD UT WOS:000085288200017 PM 10688172 ER PT J AU Corey, L Langenberg, AGM Burke, RL Straus, S AF Corey, L Langenberg, AGM Burke, RL Straus, S TI Immunologic strategies for herpes vaccination - Reply SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Letter ID PLACEBO-CONTROLLED TRIAL; SIMPLEX VIRUS TYPE-2; GENITAL HERPES; GLYCOPROTEIN-D; IMMUNOTHERAPY C1 Univ Washington, Seattle, WA 98195 USA. Chiron Corp, Emeryville, CA 94608 USA. NIH, Bethesda, MD 20892 USA. RP Corey, L (reprint author), Univ Washington, Seattle, WA 98195 USA. NR 6 TC 0 Z9 0 U1 0 U2 0 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD FEB 9 PY 2000 VL 283 IS 6 BP 746 EP 747 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 280XL UT WOS:000085129000023 ER PT J AU Waisbren, SE Hanley, W Levy, HL Shifrin, H Allred, E Azen, C Chang, PN Cipcic-Schmidt, S de la Cruz, F Hall, R Matalon, R Nanson, J Rouse, B Trefz, F Koch, R AF Waisbren, SE Hanley, W Levy, HL Shifrin, H Allred, E Azen, C Chang, PN Cipcic-Schmidt, S de la Cruz, F Hall, R Matalon, R Nanson, J Rouse, B Trefz, F Koch, R TI Outcome at age 4 years in offspring of women with maternal phenylketonuria - The maternal PKU collaborative study SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID HYPERPHENYLALANINEMIA; PREGNANCIES; ALCOHOL; PREVENTION AB Context Untreated maternal phenylketonuria (PKU) increases risk for developmental problems in offspring. The extent to which this risk is reduced by maternal dietary therapy at various stages of pregnancy is not known. Objective To determine whether dietary treatment during pregnancy of women with PKU affects developmental outcomes of offspring. Design The Maternal PKU Collaborative Study, an ongoing, longitudinal prospective study begun in 1984. Setting A total of 78 metabolic clinics and obstetrical offices in the United States, Canada, and Germany. Participants A total of 253 children of women with PKU (n = 149), with untreated mild hyperphenylalaninemia (n = 33), or without known metabolic problems (comparison group; n = 71) were followed up to age 4 years. Intervention Women with PKU were offered a low-phenylalanine diet prior to or during pregnancy with the aim of maintaining metabolic control (plasma phenylalanine less than or equal to 10 mg/dL [less than or equal to 605 mu mol/L]). Women with mild hyperphenylalaninemia, who had plasma phenylalanine levels of no more than 10 mg/dL (605 mu mol/L) on a normal diet, were not treated. Main Outcome Measures Children's scores on cognitive and behavioral assessments (McCarthy Scales of Children's Abilities, Test of Language Development, Achenbach Child Behavior Checklist, Vineland Adaptive Behavior Scales, and Home Observation for Measurement of the Environment), compared by maternal metabolic status at 0 to 10 weeks', 10 to 20 weeks', and after 20 weeks' gestation. Results Scores on the McCarthy General Cognitive Index decreased as weeks to metabolic control increased (r = -0.58; P<.001), Offspring of women who had metabolic control prior to pregnancy had a mean (SD) score of 99 (13). Forty-seven percent of offspring whose mothers did not have metabolic control by 20 weeks' gestation had a General Cognitive Index score 2 SDs below the norm. Overall, 30% of children born to mothers with PKU had social and behavioral problems. Conclusions Our data suggest that delayed development in offspring of women with PKU is associated with lack of maternal metabolic control prior to or early in pregnancy. Treatment at any time during pregnancy may reduce the severity of delay. C1 Childrens Hosp, Genet Serv, Boston, MA 02115 USA. Hosp Sick Children, Toronto, ON M5G 1X8, Canada. NICHHD, Bethesda, MD USA. Childrens Hosp Los Angeles, Los Angeles, CA 90027 USA. Childrens Hosp, Minneapolis, MN USA. Univ Tuebingen, Tuebingen, Germany. Univ Texas, Med Branch, Galveston, TX 77550 USA. Royal Univ Hosp, Saskatoon, SK S7N 0W8, Canada. Childrens Hosp Reutlingen, Reutlingen, Germany. RP Waisbren, SE (reprint author), Childrens Hosp, Genet Serv, 300 Longwood Ave,IC 107, Boston, MA 02115 USA. FU NICHD NIH HHS [N01-HD-2-3148] NR 35 TC 31 Z9 32 U1 1 U2 2 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD FEB 9 PY 2000 VL 283 IS 6 BP 756 EP 762 DI 10.1001/jama.283.6.756 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 280XL UT WOS:000085129000027 PM 10683054 ER PT J AU Seeff, LB AF Seeff, LB TI Hyman J. Zimmerman, MD - Obituary SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Biographical-Item C1 NIDDKD, NIH, Bethesda, MD 20892 USA. RP Seeff, LB (reprint author), NIDDKD, NIH, Bethesda, MD 20892 USA. NR 0 TC 1 Z9 1 U1 0 U2 0 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0098-7484 J9 JAMA-J AM MED ASSOC JI JAMA-J. Am. Med. Assoc. PD FEB 9 PY 2000 VL 283 IS 6 BP 812 EP 812 DI 10.1001/jama.283.6.812 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 280XL UT WOS:000085129000036 ER PT J AU Miller, H Prasad, R Wilson, SH Johnson, F Grollman, AP AF Miller, H Prasad, R Wilson, SH Johnson, F Grollman, AP TI 8-OxodGTP incorporation by DNA polymerase beta is modified by active-site residue Asn279 SO BIOCHEMISTRY LA English DT Article ID CRYSTAL-STRUCTURES; GAPPED DNA; FIDELITY; PROCESSIVITY; SPECIFICITY; REPLICATION; INSERTION; MECHANISM; KINETICS; DUPLEX AB To understand how the active site of a DNA polymerase might modulate the coding of 8-oxo-7,8-dihydrodeoxyguanine (8-oxodG), we performed steady-state kinetic analyses using wild-type DNA polymerase beta (pol beta) and two active-site mutants. We compared the coding of these polymerases by calculating the ratio of efficiencies for incorporation of dATP and dCTP opposite 8-oxodG and for incorporation of 8-oxodGTP opposite dA and dC. For wild-type pol beta, there is a 2:1 preference for incorporation of dCTP over dATP opposite 8-oxodG using a 5'-phosphorylated 4-base gap substrate. Mutation of either Asn279 or Arg283 to alanine has almost no effect on the ratio. 8-OxodGTP is preferentially incorporated opposite a template dA (24:1) by wild-type pol beta; mutation of Asn279 to alanine results dramatic change whereby there is preferential incorporation of 8-oxodGTP opposite dC (14:1). This suggests that interactions of 8-oxodGTP with Asn279 in the polymerase active site may alter the conformation of 8-oxodGTP and therefore alter its misincorporation. C1 SUNY Stony Brook, Dept Pharmacol Sci, Stony Brook, NY 11794 USA. Natl Inst Environm Hlth Sci, Res Triangle Pk, NC 27709 USA. RP Miller, H (reprint author), SUNY Stony Brook, Dept Pharmacol Sci, Stony Brook, NY 11794 USA. RI Miller, Holly/I-6942-2015 OI Miller, Holly/0000-0002-9076-5335 FU NCI NIH HHS [CA17395, CA47995] NR 24 TC 80 Z9 81 U1 0 U2 4 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD FEB 8 PY 2000 VL 39 IS 5 BP 1029 EP 1033 DI 10.1021/bi991789x PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 281QU UT WOS:000085172900021 PM 10653647 ER PT J AU Lenfant, C AF Lenfant, C TI NHLBI genomics initiatives - Looking beyond the Human Genome Project SO CIRCULATION LA English DT Editorial Material C1 NHLBI, Dept Hlth & Human Serv, NIH, Bethesda, MD 20892 USA. RP Lenfant, C (reprint author), NHLBI, Dept Hlth & Human Serv, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 0 TC 2 Z9 2 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD FEB 8 PY 2000 VL 101 IS 5 BP 468 EP 469 PG 2 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 281VM UT WOS:000085181500014 PM 10662740 ER PT J AU Max, MB Hagen, NA AF Max, MB Hagen, NA TI Do changes in brain sodium channels cause central pain? SO NEUROLOGY LA English DT Editorial Material C1 Natl Inst Dent & Craniofacial Res, Pain & Neurosensory Mechanisms Branch, NIH, Bethesda, MD 20892 USA. Univ Calgary, Dept Oncol, Calgary, AB T2N 1N4, Canada. Univ Calgary, Dept Clin Neurosci & Med, Calgary, AB T2N 1N4, Canada. Univ Calgary, Tom Baker Canc Ctr, Calgary, AB T2N 1N4, Canada. RP Max, MB (reprint author), Natl Inst Dent & Craniofacial Res, Pain & Neurosensory Mechanisms Branch, NIH, Bldg 10,3C-405, Bethesda, MD 20892 USA. NR 11 TC 12 Z9 13 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 FEB 8 PY 2000 VL 54 IS 3 BP 544 EP 545 PG 2 WC Clinical Neurology SC Neurosciences & Neurology GA 281KZ UT WOS:000085160300002 PM 10680780 ER PT J AU Baird, AE Lovblad, KO Schlaug, G Edelman, RR Warach, S AF Baird, AE Lovblad, KO Schlaug, G Edelman, RR Warach, S TI Multiple acute stroke syndrome - Marker of embolic disease? SO NEUROLOGY LA English DT Article; Proceedings Paper CT 50th Annual Meeting of the American-Academy-of-Neurology CY APR 25-MAY 02, 1998 CL MINNEAPOLIS, MINNESOTA SP Amer Acad Neurol DE ischemic stroke; cerebrovascular disorders; cerebral embolism ID ACUTE ISCHEMIC STROKE; BRAIN INFARCTIONS; SILENT STROKE AB Objective: To determine the frequency and etiologic significance of multiple acute ischemic lesions in stroke. Background: Although patients may have more than one stroke during the course of their lives, acute ischemic stroke is usually thought of as a single event. Using diffusion-weighted imaging (DWI), an MRI technique that detects ischemic injury within minutes after onset, we have often observed multiple acute ischemic lesions. Methods: The MRI scans of 59 consecutively studied patients were reviewed to determine the frequency and etiologic significance of multiple acute ischemic lesions on DWI. Results: Multiple acute ischemic lesions were present in 10 (17%) of 59 patients. The lesions usually occurred within one major circulation (anterior or posterior), but in two patients (3%), lesions occurred in both cerebral hemispheres or in the anterior and the posterior circulations. The lesions often were small and resulted from presumed multiple emboli or the break-up of an embolus. Two patients had internal carotid artery occlusive disease and four had a cardiac or aortic source. In the other four patients the source was not determined. Lesions larger than 1 cm in diameter progressed to infarction, but some smaller lesions were not seen on follow-up T2-weighted imaging. Conclusions: Multiple acute stroke lesions on DWI are common and could be caused by multiple emboli or the breakup of an embolus. In some cases it might become possible to make early inferences concerning the stroke mechanism that could be of use for immediately directing the clinical work-up and treatment of the patient. C1 Beth Israel Deaconess Med Ctr, Dept Neurol, Boston, MA 02215 USA. Beth Israel Deaconess Med Ctr, Dept Radiol, Boston, MA 02215 USA. Harvard Univ, Sch Med, Boston, MA USA. NINDS, Div Diagnost & Therapeut, Bethesda, MD 20892 USA. RP Baird, AE (reprint author), Beth Israel Deaconess Med Ctr, Dept Neurol, DANA 779,E Campus, Boston, MA 02215 USA. NR 24 TC 107 Z9 118 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 FEB 8 PY 2000 VL 54 IS 3 BP 674 EP 678 PG 5 WC Clinical Neurology SC Neurosciences & Neurology GA 281KZ UT WOS:000085160300024 PM 10680802 ER PT J AU Power, C Gladden, JGB Halliday, W Del Bigio, MR Nath, A Ni, W Major, EO Blanchard, J Mowat, M AF Power, C Gladden, JGB Halliday, W Del Bigio, MR Nath, A Ni, W Major, EO Blanchard, J Mowat, M TI AIDS- and non-AIDS-related PML association with distinct p53 polymorphism SO NEUROLOGY LA English DT Article DE progressive multifocal leukoencephalopathy; AIDS; p53 polymorphism ID PROGRESSIVE MULTIFOCAL LEUKOENCEPHALOPATHY; CANCER AB A population-based analysis of progressive multifocal leukoencephalopathy (PML) showed PML frequencies of 5.1% among patients with AIDS and 0.07% among patients with hematologic malignancies, but similar clinical features of PML in both groups. Sequencing of the p53 gene, exon 4, showed heterozygosity (Arg-Pro) at codon 72 in five of six PML patients. These findings indicate that frequencies of non-AIDS- and AIDS-related PML differ markedly but p53 polymorphisms may influence the occurrence of PML in both groups. C1 Univ Calgary, Dept Clin Neurol, Calgary, AB T2N 4N1, Canada. Univ Manitoba, Dept Pharmacol & Therapeut, Winnipeg, MB, Canada. Univ Manitoba, Dept Pathol, Winnipeg, MB, Canada. Univ Manitoba, Dept Med Microbiol, Winnipeg, MB, Canada. Manitoba Inst Cell Biol, Winnipeg, MB R3E 0V9, Canada. Univ Kentucky, Dept Neurol Microbiol & Immunol, Lexington, KY USA. NINDS, Lab Mol Med & Neurosci, Bethesda, MD 20892 USA. RP Power, C (reprint author), Univ Calgary, Dept Clin Neurol, Calgary, AB T2N 4N1, Canada. RI Power, Christopher/C-7181-2013; OI Power, Christopher/0000-0002-5131-9711; Del Bigio, Marc/0000-0001-8366-0394 NR 10 TC 44 Z9 45 U1 1 U2 3 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0028-3878 J9 NEUROLOGY JI Neurology PD FEB 8 PY 2000 VL 54 IS 3 BP 743 EP 746 PG 4 WC Clinical Neurology SC Neurosciences & Neurology GA 281KZ UT WOS:000085160300038 PM 10680816 ER PT J AU Foroud, T Castelluccio, PF Koller, DL Edenberg, HJ Miller, M Bowman, E Rau, NL Smiley, C Rice, JP Goate, A Armstrong, C Bierut, LJ Reich, T Detera-Wadleigh, SD Goldin, LR Badner, JA Guroff, JJ Gershon, ES McMahon, FJ Simpson, S MacKinnon, D McInnis, M Stine, OC DePaulo, JR Blehar, MC Nurnberger, JI AF Foroud, T Castelluccio, PF Koller, DL Edenberg, HJ Miller, M Bowman, E Rau, NL Smiley, C Rice, JP Goate, A Armstrong, C Bierut, LJ Reich, T Detera-Wadleigh, SD Goldin, LR Badner, JA Guroff, JJ Gershon, ES McMahon, FJ Simpson, S MacKinnon, D McInnis, M Stine, OC DePaulo, JR Blehar, MC Nurnberger, JI TI Suggestive evidence of a locus on chromosome 10p using the NIMH genetics initiative bipolar affective disorder pedigrees SO AMERICAN JOURNAL OF MEDICAL GENETICS LA English DT Article DE bipolar affective disorder; genetics; linkage; nonparametric analysis; genomic survey ID NONPARAMETRIC LINKAGE ANALYSIS; MANIC-DEPRESSIVE ILLNESS; SUSCEPTIBILITY LOCUS; GENOME SCAN; CONTROLLED FAMILY; DNA MARKERS; SCHIZOPHRENIA; SUPPORT; REGION AB As part of a four-center NIMH Genetics Initiative on Bipolar Disorder, a genome screen using 365 markers was performed on 540 DNAs from 97 families, enriched for affected relative pairs. This is the largest uniformly ascertained and assessed linkage sample for this disease, and includes 232 subjects diagnosed with bipolar I (BPI), 32 with schizo-affective, bipolar type (SABP), 72 with bipolar II (DPD), and 88 with unipolar recurrent depression (UPR). A hierarchical set of definitions of affected status was examined. Under Model I, affected individuals were those with a diagnosis of BPI or SABP, Model PI included as affected those fitting Model I plus BPII, and Model LII included those fitting Model II plus UPR. This data set was previously analyzed using primarily affected sib pair methods. We report the results of nonparametric linkage analyses of the extended pedigree structure using the program Genehunter Plus. The strongest finding was a lod score of 2.5 obtained on chromosome 10 near the marker D10S1423 with diagnosis as defined under Model II, This region has been previously implicated in genome-wide studies of schizophrenia and bipolar disorder. Other chromosomal regions with lod scores over 1.50 for at least one Model Included chromosomes 8 (Model III), 16 (Model III), and 20 (Model I). Am. J. Med. Genet. (Neuropsychiatr, Genet.) 96:18-23, 2000 (C) 2000 Wiley-Liss, Inc. C1 Indiana Univ, Sch Med, Dept Med & Mol Genet, Indianapolis, IN 46202 USA. Washington Univ, Sch Med, Dept Psychiat, St Louis, MO 63110 USA. NIMH, Clin Neurogenet Branch, NIH, Bethesda, MD 20892 USA. Johns Hopkins Univ, Sch Med, Dept Psychiat, Baltimore, MD 21205 USA. NIMH, Mood Anxiety & Personal Disorders Res Branch, NIH, Rockville, MD 20857 USA. Indiana Univ, Sch Med, Dept Psychiat, Indianapolis, IN 46202 USA. RP Foroud, T (reprint author), Indiana Univ, Sch Med, Dept Med & Mol Genet, 975 W Walnut St IB 155, Indianapolis, IN 46202 USA. RI McMahon, Francis/A-7290-2009; OI Nurnberger, John/0000-0002-7674-1767; McMahon, Francis/0000-0002-9469-305X; Edenberg, Howard/0000-0003-0344-9690 FU NIMH NIH HHS [U01 MH46280, U01 MH46282, U01 MH54794] NR 40 TC 57 Z9 57 U1 2 U2 2 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0148-7299 J9 AM J MED GENET JI Am. J. Med. Genet. PD FEB 7 PY 2000 VL 96 IS 1 BP 18 EP 23 DI 10.1002/(SICI)1096-8628(20000207)96:1<18::AID-AJMG6>3.0.CO;2-G PG 6 WC Genetics & Heredity SC Genetics & Heredity GA 276TM UT WOS:000084893500006 PM 10686547 ER PT J AU Taktakishvili, M Neamati, N Pommier, Y Nair, V AF Taktakishvili, M Neamati, N Pommier, Y Nair, V TI Recognition and inhibition of HIV integrase by a novel dinucleotide SO BIOORGANIC & MEDICINAL CHEMISTRY LETTERS LA English DT Article DE antivirals; nucleotides; enzyme inhibitors ID IMMUNODEFICIENCY-VIRUS TYPE-1; ISOMERIC DIDEOXYNUCLEOSIDES; ANTIVIRAL AGENTS; VIRAL-DNA AB The viral enzyme, HIV integrase, is involved in the integration of viral DNA into host cell DNA. In the quest for a small nucleotide system with nuclease stability of the internucleotide phosphate bond and critical structural features for recognition and inhibition of HIV-1 integrase, we have discovered a conceptually novel dinucleotide, pIsodApdC, which is a potent inhibitor of this key viral enzyme. (C) 2000 Elsevier Science Ltd. All rights reserved. C1 Univ Iowa, Dept Chem, Iowa City, IA 52242 USA. NCI, Pharmacol Lab, NIH, Bethesda, MD 20892 USA. RP Nair, V (reprint author), Univ Iowa, Dept Chem, Iowa City, IA 52242 USA. FU NIAID NIH HHS [AI 43181] NR 17 TC 14 Z9 15 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0960-894X J9 BIOORG MED CHEM LETT JI Bioorg. Med. Chem. Lett. PD FEB 7 PY 2000 VL 10 IS 3 BP 249 EP 251 DI 10.1016/S0960-894X(99)00677-0 PG 3 WC Chemistry, Medicinal; Chemistry, Organic SC Pharmacology & Pharmacy; Chemistry GA 284WJ UT WOS:000085355600012 PM 10698446 ER PT J AU Sijts, AJAM Ruppert, T Rehermann, B Schmidt, M Koszinowski, U Kloetzel, PM AF Sijts, AJAM Ruppert, T Rehermann, B Schmidt, M Koszinowski, U Kloetzel, PM TI Efficient generation of a hepatitis B virus cytotoxic T lymphocyte epitope requires the structural features of immunoproteasomes SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Article DE antigen processing; LMP7T1A; hepatitis B virus; immunoproteasome; MHC class I ID MAJOR HISTOCOMPATIBILITY COMPLEX; MHC CLASS-I; MULTICATALYTIC PROTEINASE COMPLEX; PROTEASOME BETA-SUBUNITS; ACUTE VIRAL-HEPATITIS; ANTIGEN PRESENTATION; INTERFERON-GAMMA; 20S PROTEASOME; ENCODED SUBUNITS; IFN-GAMMA AB Interferon (IFN)-gamma-induced cells express the proteasome subunits low molecular weight protein (LMP)2, LMP7, and MECL-1 (multicatalytic endopeptidase complex-like 1), leading to the formation of immunoproteasomes. Although these subunits are thought to optimize MHC class I antigen processing, the extent of their role and the mechanistic aspects involved remain unclear. Herein, we study the proteolytic generation of an human histocompatibility leukocyte antigen (HLA)-Aw68-restricted hepatitis B virus core antigen (HBcAg) cytotoxic T lymphocyte (CTL) epitope that is recognized by peripheral blood lymphocytes from patients with acute self-limited but not chronic hepatitis B virus (HBV). Immunological data suggest that IFN-gamma-induced rather than uninduced HeLa cells process and present the HBV CTL epitope upon infection with HBcAg-expressing vaccinia viruses. Analyses of 20S proteasome digests of. synthetic polypeptides coveting the antigenic HBcAg peptide demonstrate that only immunoproteasomes efficiently perform the cleavages needed for the liberation of this HBV CTL epitope. Although the concerted presence of the three immunosubunits appears essential, we find that both catalytically active LMP7 and inactive LMP7 T1A support CTL epitope generation. We conclude that LMP7 influences the structural features of 20S proteasomes, thereby enhancing the activity of the LMP2 and MECL-1 catalytic sites, which provide cleavage specificity. Thus, LMP7 incorporation is of greater functional importance for the generation of an HBV CTL epitope than cleavage specificity. C1 Humboldt Univ, Charite, Inst Biochem, D-10117 Berlin, Germany. Max Von Pettenkofer Inst, D-80336 Munich, Germany. NIDDKD, Liver Dis Sect, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. RP Kloetzel, PM (reprint author), Humboldt Univ, Charite, Inst Biochem, Monbijoustr 2, D-10117 Berlin, Germany. NR 54 TC 109 Z9 114 U1 0 U2 1 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 FEB 7 PY 2000 VL 191 IS 3 BP 503 EP 513 DI 10.1084/jem.191.3.503 PG 11 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 283UW UT WOS:000085295300011 PM 10662796 ER PT J AU Benyassi, A Schwartz, C Coon, SL Klein, DC Falcon, J AF Benyassi, A Schwartz, C Coon, SL Klein, DC Falcon, J TI Melatonin synthesis: arylalkylamine N-acetyltransferases in trout retina and pineal organ are different SO NEUROREPORT LA English DT Article DE arylalkylamine N-acetyltransferase; fish; pineal organ; retina AB Serotonin N-acetyltransferase (AANAT) is the first enzyme in the conversion of serotonin to melatonin. Changes in AANAT activity determine the daily rhythm in melatonin secretion. Two AANAT genes have been identified in the pike, pAANAT-1 and pAANAT-2, expressed in the retina and in the pineal, respectively. The genes preferentially expressed in these tissues encode proteins with distinctly different kinetic characteristics. Like the pike, trout retina primarily expresses the AANAT-1 gene and trout pineal primarily expresses the AANAT-2 gene. Here we show that the kinetic characteristics of AANAT in these tissues differ as in pike. These differences include optimal temperature for activity (pineal: 12 degrees C; retina: 25 degrees C) and relative affinity for indoleethylamines compared to phenylethylamines. In addition, retinal AANAT exhibited substrate inhibition, which was not seen with pineal AANAT. The kinetic differences between AANAT-1 and AANAT-2 appear to be defining characteristics of these gene subfamilies, and are not species specific. NeuroReport 11:255-258 (C) 2000 Lippincott Williams & Wilkins. C1 NICHHD, Sect Neuroendocrinol, Dev Neurobiol Lab, NIH, Bethesda, MD 20892 USA. Univ Poitiers, CNRS, UMR 6558, Lab Neurobiol Cellulaire & Neuroendocrinol, F-86022 Poitiers, France. Fac Sci Fes, Physiol Anim Lab, Fes, Morocco. RP Falcon, J (reprint author), NICHHD, Sect Neuroendocrinol, Dev Neurobiol Lab, NIH, Bethesda, MD 20892 USA. RI FALCON, Jack/I-5302-2013 OI FALCON, Jack/0000-0002-7572-6581 NR 9 TC 40 Z9 42 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0959-4965 J9 NEUROREPORT JI Neuroreport PD FEB 7 PY 2000 VL 11 IS 2 BP 255 EP 258 DI 10.1097/00001756-200002070-00006 PG 4 WC Neurosciences SC Neurosciences & Neurology GA 278LX UT WOS:000084992200007 PM 10674465 ER PT J AU Zhang, L Zhao, WQ Li, BS Alkon, DL Barker, JL Chang, YH Wu, M Rubinow, DR AF Zhang, L Zhao, WQ Li, BS Alkon, DL Barker, JL Chang, YH Wu, M Rubinow, DR TI TNF-alpha induced over-expression of GFAP is associated with MAPKs SO NEUROREPORT LA English DT Article DE astrocyte; GFAP; MAPKs; tumor necrosis factor-alpha ID TUMOR-NECROSIS-FACTOR; FIBRILLARY ACIDIC PROTEIN; CREUTZFELDT-JAKOB-DISEASE; CULTURED ASTROCYTES; BRAIN IRRADIATION; CELLS; CYTOKINES; MICE; INTERLEUKIN-1-ALPHA; ASTROGLIOSIS AB Increased levels of tumor necrosis factor-alpha (TNF-alpha), a pluripotent cytokine that is reportedly mitogenic to astrocytes, are associated with the expression of glial fibrillary acidic protein (GFAP), the most specific marker for astrocytes, in many neuropathological conditions, including brain injury, CNS infection, Creutzfeldt-Jakob disease and Alzheimer's disease. Here, we show that treatment of cultured astrocytes with TNF-alpha resulted in dramatic over-expression of GFAP, associated with a substantial activation of the mitogen activated protein kinase (MAPK) Erk2 (extracellular signal-regulated protein kinase). We also demonstrate chat TNF-alpha-induced over-expression of GFAP was significantly attenuated by the MAPK inhibitor PD98059. We conclude that TNF-alpha may upregulate GFAP through the MAPK signaling pathway. Because increased GFAP is a hallmark of reactive gliosis, understanding the mechanisms that regulate GFAP expression may facilitate development of strategies to minimize the gliosis associated with many brain diseases. NeuroReport 11:409-412 (C) 2000 Lippincott Williams & Wilkins. C1 NIMH, Behav Endocrinol Branch, NIH, Bethesda, MD 20892 USA. NINDS, Lab Adapt Syst, NIH, Bethesda, MD 20892 USA. NINDS, Neurochem Lab, NIH, Bethesda, MD 20892 USA. NINDS, Neurophysiol Lab, NIH, Bethesda, MD 20892 USA. RP Zhang, L (reprint author), NIMH, Behav Endocrinol Branch, NIH, Bldg 36,2C02,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 23 TC 33 Z9 36 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0959-4965 J9 NEUROREPORT JI Neuroreport PD FEB 7 PY 2000 VL 11 IS 2 BP 409 EP 412 DI 10.1097/00001756-200002070-00037 PG 4 WC Neurosciences SC Neurosciences & Neurology GA 278LX UT WOS:000084992200038 PM 10674496 ER PT J AU Kruger, EA Duray, PH Tsokos, MG Venzon, DJ Libutti, SK Dixon, SC Rudek, MA Pluda, J Allegra, C Figg, WD AF Kruger, EA Duray, PH Tsokos, MG Venzon, DJ Libutti, SK Dixon, SC Rudek, MA Pluda, J Allegra, C Figg, WD TI Endostatin inhibits microvessel formation in the ex vivo rat aortic ring angiogenesis assay SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article ID COLLAGEN; CANCER; XVIII AB Endostatin has demonstrated potent antiangiogenic and antitumor activity in mouse models. We have investigated the ex vivo rat aortic ring assay and a human vein model to assess the biological activity of murine and human endostatin. Rat aortic rings were exposed to recombinant murine endostatin (Spodoptera frugipera; Calbiochem, San Diego, CA) or recombinant human endostatin (Pichia pastoris; EntreMed, Rockville, MD). After 5 days, murine endostatin (500 mu g/ml) demonstrated inhibition of microvessel outgrowth with dose-dependent effects (down to 18 mu g/ml). No significant inhibition was observed with human endostatin in the rat assay. Human endostatin at 250 and 500 mu g/ml inhibited outgrowths from human saphenous vein rings after a 14-day incubation. Electron microscopy assessed the formation of basal lamina, confirming that the microvessels were progenitors of patent vessels. Immunostaining for Factor VIII or CD34 demonstrated that the microvessel cells were endothelial. BrdU incorporation assays supported the presence of proliferating endothelial cells, correlating with neovascularization from the aortic wall. We conclude that the rat aortic ring assay confirms the antiangiogenic activity of murine but not human endostatin, suggesting that the model may have species specificity. However, the human form shows biological activity against human vascular tissue, (C) 2000 Academic Press. C1 NCI, Med Branch, Div Clin Sci, Bethesda, MD 20892 USA. NCI, DCS, Pathol Lab, Bethesda, MD 20892 USA. NCI, DCS, Biostat & Data Management Sect, Bethesda, MD 20892 USA. NCI, DCS, Surg Branch, Bethesda, MD 20892 USA. NCI, Canc Treatment & Evaluat Program, Bethesda, MD 20892 USA. RP Figg, WD (reprint author), NIH, Bldg 10,Room 5A01, Bethesda, MD 20892 USA. RI Venzon, David/B-3078-2008; Figg Sr, William/M-2411-2016 NR 19 TC 69 Z9 76 U1 0 U2 3 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD FEB 5 PY 2000 VL 268 IS 1 BP 183 EP 191 DI 10.1006/bbrc.1999.2018 PG 9 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 281RA UT WOS:000085173500034 PM 10652234 ER PT J AU Hosohata, Y Vanderah, TW Burkey, TH Ossipov, MH Kovelowski, CJ Sora, I Uhl, GR Zhang, XY Rice, KC Roeske, WR Hruby, VJ Yamamura, HI Lai, J Porreca, F AF Hosohata, Y Vanderah, TW Burkey, TH Ossipov, MH Kovelowski, CJ Sora, I Uhl, GR Zhang, XY Rice, KC Roeske, WR Hruby, VJ Yamamura, HI Lai, J Porreca, F TI delta-opioid receptor agonists produce antinociception and [S-35]GTP gamma S binding in mu receptor knockout mice SO EUROPEAN JOURNAL OF PHARMACOLOGY LA English DT Article DE delta-opioid receptor; mu-opioid receptor; mu-opioid receptor-knockout mice; antinociception; G protein; opioid drug ID MORPHINE-INDUCED ANALGESIA; ANTISENSE OLIGODEOXYNUCLEOTIDE; SPINAL ANTINOCICEPTION; D-PEN5>ENKEPHALIN; EFFICACY; LACKING; 3.0.CO;2-8 PG 13 WC Zoology SC Zoology GA 277NW UT WOS:000084940500001 PM 10676707 ER PT J AU Rapoport, JL AF Rapoport, JL TI The development of neurodevelopmental psychiatry SO AMERICAN JOURNAL OF PSYCHIATRY LA English DT Editorial Material ID CEREBRAL-PALSY; FETAL GROWTH; COHORT; RISK C1 NIMH, Child Psychiat Branch, Bethesda, MD 20892 USA. RP Rapoport, JL (reprint author), Bldg 10,Rm 6N-240,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 10 TC 7 Z9 7 U1 0 U2 0 PU AMER PSYCHIATRIC PRESS, INC PI WASHINGTON PA 1400 K ST, N W, STE 1101, WASHINGTON, DC 20005 USA SN 0002-953X J9 AM J PSYCHIAT JI Am. J. Psychiat. PD FEB PY 2000 VL 157 IS 2 BP 159 EP 161 PG 3 WC Psychiatry SC Psychiatry GA 281PC UT WOS:000085169000001 ER PT J AU McNeil, TF Cantor-Graae, E Weinberger, DR AF McNeil, TF Cantor-Graae, E Weinberger, DR TI Relationship of obstetric complications and differences in size of brain structures in monozygotic twin pairs discordant for schizophrenia SO AMERICAN JOURNAL OF PSYCHIATRY LA English DT Article ID NEUROLOGICAL ABNORMALITIES; CORTICAL DEVELOPMENT; TEMPORAL-LOBE; HIGH-RISK; DISORDER; NEUROPATHOLOGY; DISTURBANCES; HIPPOCAMPUS; POSTMORTEM; CONCORDANT AB Objective: The aim of the study was to determine whether a history of obstetric complications and congenital minor physical anomalies are related to differences in the characteristics of brain structures observed within monozygotic twin pairs discordant for schizophrenia. Method: The size of the bilateral hippocampi and cerebral ventricles was studied by magnetic resonance imaging in 22 monozygotic twin pairs discordant for schizophrenia. Obstetric complications and minor physical anomalies were independently assessed through parental report and examination, respectively. Results: Compared with the well co-twins, the ill twins consistently had smaller left and right hippocampi as well as larger left lateral ventricles and third ventricles, Relatively small left and right hippocampi were each significantly related to labor-delivery complications and to prolonged labor per se. Relatively large right lateral ventricle size and large total ventricle size were significantly related to labor-delivery complications, prolonged labor, neonatal complications, and total complications for the entire reproductive sequence. In contrast, these brain size differences were not significantly associated with pregnancy complications or minor physical anomalies. Conclusions: Trauma at the time of labor and delivery and especially prolonged labor appear to be of importance for brain structure anomalies associated with schizophrenia. C1 Univ Lund, Dept Community Med, Hosp MAS, S-20502 Malmo, Sweden. NIMH, Clin Brain Disorders Branch, Bethesda, MD 20892 USA. RP McNeil, TF (reprint author), Univ Lund, Dept Community Med, Hosp MAS, S-20502 Malmo, Sweden. FU NIMH NIH HHS [MH-41176] NR 63 TC 126 Z9 129 U1 5 U2 12 PU AMER PSYCHIATRIC PRESS, INC PI WASHINGTON PA 1400 K ST, N W, STE 1101, WASHINGTON, DC 20005 USA SN 0002-953X J9 AM J PSYCHIAT JI Am. J. Psychiat. PD FEB PY 2000 VL 157 IS 2 BP 203 EP 212 DI 10.1176/appi.ajp.157.2.203 PG 10 WC Psychiatry SC Psychiatry GA 281PC UT WOS:000085169000009 PM 10671388 ER PT J AU Solomon, DA Keller, MB Leon, AC Mueller, TI Lavori, PW Shea, T Coryell, W Warshaw, M Turvey, C Maser, JD Endicott, J AF Solomon, DA Keller, MB Leon, AC Mueller, TI Lavori, PW Shea, T Coryell, W Warshaw, M Turvey, C Maser, JD Endicott, J TI Multiple recurrences of major depressive disorder SO AMERICAN JOURNAL OF PSYCHIATRY LA English DT Article ID DOUBLE-BLIND; UNIPOLAR; RELAPSE; CONTINUATION; PREVENTION; THERAPY; LITHIUM AB Objective: The authors of this study examined multiple recurrences of unipolar major depressive disorder. Method: A total of 318 subjects with unipolar major depressive disorder were prospectively followed for 10 years within a multicenter naturalistic study. Survival analytic techniques were used to examine the probability of recurrence after recovery from the index episode. Results: The mean number of episodes of major depression per year of follow-up was 0.21, and nearly two-thirds of the subjects suffered at least one recurrence. The number of lifetime episodes of major depression was significantly associated with the probability of recurrence, such that the risk of recurrence increased by 16% with each successive recurrence. The risk of recurrence progressively decreased as the duration of recovery increased. Within subjects, there was very little consistency in the time to recurrence. Conclusions: Major depressive disorder is a highly recurrent illness. The risk of the recurrence of major depressive disorder progressively increases with each successive episode and decreases as the duration of recovery increases. C1 Rhode Isl Hosp, Dept Psychiat, Mood Disorders Program, Providence, RI 02903 USA. Brown Univ, Dept Psychiat & Human Behav, Providence, RI 02912 USA. Cornell Univ, Dept Psychiat, New York, NY USA. Stanford Univ, Dept Hlth Policy & Res, Stanford, CA 94305 USA. Univ Iowa, Coll Med, Dept Psychiat, Iowa City, IA 52242 USA. NIMH, Brain Disorders Program, Bethesda, MD 20892 USA. New York State Psychiat Inst, Dept Res & Training, New York, NY 10032 USA. RP Solomon, DA (reprint author), Rhode Isl Hosp, Dept Psychiat, Mood Disorders Program, 593 Eddy St, Providence, RI 02903 USA. FU NIMH NIH HHS [R01 MH025478] NR 21 TC 335 Z9 344 U1 12 U2 44 PU AMER PSYCHIATRIC PRESS, INC PI WASHINGTON PA 1400 K ST, N W, STE 1101, WASHINGTON, DC 20005 USA SN 0002-953X J9 AM J PSYCHIAT JI Am. J. Psychiat. PD FEB PY 2000 VL 157 IS 2 BP 229 EP 233 DI 10.1176/appi.ajp.157.2.229 PG 5 WC Psychiatry SC Psychiatry GA 281PC UT WOS:000085169000012 PM 10671391 ER PT J AU Sherbourne, CD Hays, RD Fleishman, JA Vitiello, B Magruder, KM Bing, EG McCaffrey, D Burnam, A Longshore, D Eggan, F Bozzette, SA Shapiro, MF AF Sherbourne, CD Hays, RD Fleishman, JA Vitiello, B Magruder, KM Bing, EG McCaffrey, D Burnam, A Longshore, D Eggan, F Bozzette, SA Shapiro, MF TI Impact of psychiatric conditions on health-related quality of life in persons with HIV infection SO AMERICAN JOURNAL OF PSYCHIATRY LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; OF-LIFE; DISORDERS; PREVALENCE; DEPRESSION; DISEASE; MEN; RELIABILITY; DISABILITY; VALIDITY AB Objective: Little is known about the impact of comorbid psychiatric symptoms in persons with HIV. This study estimates the burden on health-related quality of life associated with comorbid psychiatric conditions in a nationally representative sample of persons with HIV. Method: The authors conducted a multistage sampling of urban and rural areas to produce a national probability sample of persons with HIV receiving medical care in the contiguous United States (N=2,864). Subjects were screened for psychiatric conditions with the short form of the Composite International Diagnostic interview. Heavy drinking was assessed on the basis of quantity and frequency of drinking. Health-related quality of life was rated with a 28-item instrument adapted from similar measures used in the Medical Outcomes Study. Results: HIV subjects with a probable mood disorder diagnosis had significantly lower scores on health-related quality of life measures than did those without such symptoms. Diminished health-related quality of life was not associated with heavy drinking, and in drug users it was accounted for by presence of a comoribid mood disorder. Conclusions: Optimization of health-related quality of life is particularly important now that HIV is a chronic disease with the prospect of long-term survival. Comorbid psychiatric conditions may serve as markers for impaired functioning and well-being in persons with HIV, Inclusion of sufficient numbers of appropriately trained mental health professionals to identify and treat such conditions may reduce unnecessary utilization of other health services and improve health-related quality of life in persons with HIV infection. C1 RAND Corp, Hlth Div, Santa Monica, CA 90407 USA. NIMH, Rockville, MD 20857 USA. Agcy Healthcare Res & Qual, Rockville, MD 20857 USA. RP Sherbourne, CD (reprint author), RAND Corp, Hlth Div, 1700 Main St,Box 2138, Santa Monica, CA 90407 USA. RI Hays, Ronald/D-5629-2013 FU AHRQ HHS [HS-08578] NR 35 TC 151 Z9 157 U1 1 U2 8 PU AMER PSYCHIATRIC PRESS, INC PI WASHINGTON PA 1400 K ST, N W, STE 1101, WASHINGTON, DC 20005 USA SN 0002-953X J9 AM J PSYCHIAT JI Am. J. Psychiat. PD FEB PY 2000 VL 157 IS 2 BP 248 EP 254 DI 10.1176/appi.ajp.157.2.248 PG 7 WC Psychiatry SC Psychiatry GA 281PC UT WOS:000085169000016 PM 10671395 ER PT J AU Giedd, JN Rapoport, JL Garvey, MA Perlmutter, S Swedo, SE AF Giedd, JN Rapoport, JL Garvey, MA Perlmutter, S Swedo, SE TI MRI assessment of children with obsessive-compulsive disorder or tics associated with streptococcal infection SO AMERICAN JOURNAL OF PSYCHIATRY LA English DT Article AB Objective: The authors assessed selective basal ganglia involvement in a subgroup of children with obsessive-compulsive disorder (OCD) and/or ties believed to be associated with streptococcal infection. Method: Using computer-assisted morphometric techniques, they analyzed the cerebral magnetic resonance images of 34 children with presumed streptococcus-associated OCD and/or ties and 82 healthy comparison children who were matched for age and sex. Results: The average sizes of the caudate, putamen, and globus pallidus, but not of the thalamus or total cerebrum, were significantly greater in the group of children with streptococcus-associated OCD and/or ties than in the healthy children. The differences were similar to those found previously for subjects with Sydenham's chorea compared with normal subjects. Conclusions: These results support the hypothesis that there is a distinct subgroup of subjects with OCD and/or ties who have enlarged basal ganglia, These findings are consistent with the hypothesis of an autoimmune response to streptococcal infection. C1 NIMH, Child Psychiat Branch, Bethesda, MD 20892 USA. NIMH, Pediat Branch, Bethesda, MD 20892 USA. NIMH, Dev Neuropsychiat Branch, Bethesda, MD 20892 USA. RP Giedd, JN (reprint author), NIMH, Child Psychiat Branch, 10 Ctr Dr,MSC 1367,Bldg 10,Rm 4C110, Bethesda, MD 20892 USA. RI Giedd, Jay/A-3080-2008; Giedd, Jay/B-7302-2012; Giedd, Jay/J-9644-2015 OI Giedd, Jay/0000-0003-0827-3460; Giedd, Jay/0000-0003-2002-8978 NR 5 TC 145 Z9 149 U1 1 U2 5 PU AMER PSYCHIATRIC PRESS, INC PI WASHINGTON PA 1400 K ST, N W, STE 1101, WASHINGTON, DC 20005 USA SN 0002-953X J9 AM J PSYCHIAT JI Am. J. Psychiat. PD FEB PY 2000 VL 157 IS 2 BP 281 EP 283 DI 10.1176/appi.ajp.157.2.281 PG 3 WC Psychiatry SC Psychiatry GA 281PC UT WOS:000085169000024 PM 10671403 ER PT J AU Fee, E Brown, TM AF Fee, E Brown, TM TI The microbial menace, then and now SO AMERICAN JOURNAL OF PUBLIC HEALTH LA English DT Editorial Material C1 Natl Lib Med, Hist Med Div, Bethesda, MD 20894 USA. Univ Rochester, Med Ctr, Dept Community & Prevent Med, Rochester, NY 14642 USA. RP Fee, E (reprint author), Natl Lib Med, Hist Med Div, Bldg 38,Room 1E21,8600 Rockville Pike, Bethesda, MD 20894 USA. NR 5 TC 2 Z9 2 U1 0 U2 0 PU AMER PUBLIC HEALTH ASSOC INC PI WASHINGTON PA 1015 FIFTEENTH ST NW, WASHINGTON, DC 20005 USA SN 0090-0036 J9 AM J PUBLIC HEALTH JI Am. J. Public Health PD FEB PY 2000 VL 90 IS 2 BP 184 EP 185 DI 10.2105/AJPH.90.2.184 PG 2 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 278QA UT WOS:000084999400004 PM 10667176 ER PT J AU Artz, L Macaluso, M Brill, I Kelaghan, J Austin, H Fleenor, M Robey, L Hook, EW AF Artz, L Macaluso, M Brill, I Kelaghan, J Austin, H Fleenor, M Robey, L Hook, EW TI Effectiveness of an intervention promoting the female condom to patients at sexually transmitted disease clinics SO AMERICAN JOURNAL OF PUBLIC HEALTH LA English DT Article ID AFRICAN-AMERICAN WOMEN; ACCEPTABILITY; PREVENTION; INFECTION; RISK AB Objectives. This study evaluated a behavioral intervention designed to promote female condoms and reduce unprotected sex among women at high risk for acquiring sexually transmitted diseases (STDs). Methods. The effect of the intervention on barrier use was evaluated with a pretest-posttest design with 1159 female STD clinic patients. Results. Among participants with follow-up data, 79% used the female condom at least once and after multiple times. More than one third of those who completed the study used female condoms throughout follow-up. Use of barrier protection increased significantly after the intervention, and high use was maintained during a 6-month follow-up. To account for attrition, the use of protection by all subjects was projected under 3 conservative assumptions. The initial visit and termination visit projections suggest that use increased sharply after the intervention and declined during follow-up but remained elevated compared with the baseline. Conclusions. Many clients of public STD clinics will try, and some will continue, to use female condoms when they are promoted positively and when women are trained to use them correctly and to promote them to their partners. A behavioral intervention that promotes both female and male condoms can increase barrier use. C1 Univ Alabama, Sch Publ Hlth, Dept Epidemiol & Int Hlth, Birmingham, AL 35294 USA. NICHHD, Contracept & Reprod Evaluat Branch, Bethesda, MD 20892 USA. Emory Univ, Dept Epidemiol, Rollins Sch Publ Hlth, Atlanta, GA 30322 USA. Jefferson Cty Dept Hlth, Birmingham, AL USA. Madison Cty Hlth Dept, Huntsville, AL USA. Univ Alabama, Sch Med, Div Infect Dis, Birmingham, AL USA. RP Macaluso, M (reprint author), Univ Alabama, Sch Publ Hlth, Dept Epidemiol & Int Hlth, MJH 108,1530 3rd Ave S, Birmingham, AL 35294 USA. RI Macaluso, Maurizio/J-2076-2015 OI Macaluso, Maurizio/0000-0002-2977-9690 FU NICHD NIH HHS [N01-HD-1-3135]; PHS HHS [U48/CCU409679-02] NR 30 TC 73 Z9 73 U1 0 U2 4 PU AMER PUBLIC HEALTH ASSOC INC PI WASHINGTON PA 1015 FIFTEENTH ST NW, WASHINGTON, DC 20005 USA SN 0090-0036 J9 AM J PUBLIC HEALTH JI Am. J. Public Health PD FEB PY 2000 VL 90 IS 2 BP 237 EP 244 DI 10.2105/AJPH.90.2.237 PG 8 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 278QA UT WOS:000084999400013 PM 10667185 ER PT J AU Kogan, MD Overpeck, MD Hoffman, HJ Casselbrant, ML AF Kogan, MD Overpeck, MD Hoffman, HJ Casselbrant, ML TI Factors associated with tympanostomy tube insertion among preschool-aged children in the United States SO AMERICAN JOURNAL OF PUBLIC HEALTH LA English DT Article ID ACUTE OTITIS-MEDIA; ENVIRONMENTAL TOBACCO-SMOKE; MIDDLE-EAR DISEASE; HEALTH-INSURANCE; YOUNG-CHILDREN; RISK-FACTORS; CARE; EFFUSION; APPROPRIATENESS; PREVALENCE AB Objective. Recurrent and persistent otitis media is often treated by tympanostomy tube insertion to ventilation the middle ear and restore hearing. This study examined the factors that predict which children are most likely to receive tympanostomy tubes through 3 years of age. Methods. Multiple logistic regression was conducted on data from a nationally representative sample of children (N = 8285). Results. By 3 years of age 6.8% of US children has tubes inserted. Logistic regression indicated that after control for number of ear infections, children without any gaps in health insurance, who attended a day-care center, who were White, whose birth-weight was less than 1500 g, who lived in the Midwest or South were significantly more likely to have tympanostomy tubes. Conclusions. These data suggest that differences exist as to who receives tubes. Of particular concern are differences by race/ethnicity and continuity of health care coverage. With expansions in health care coverage to larger proportions of uninsured children, it will be important to monitor these programs to ensure that all children who may need tympanostomy tubes have access to them. C1 Ctr Dis Control & Prevent, Natl Ctr Hlth Stat, Hyattsville, MD 20782 USA. NICHHD, NIH, Bethesda, MD 20892 USA. NIDOCD, NIH, Bethesda, MD USA. Univ Pittsburgh, Sch Med, Dept Otolaryngol, Pittsburgh, PA 15260 USA. Childrens Hosp Pittsburgh, Dept Pediat Otolaryngol, Pittsburgh, PA 15213 USA. RP Kogan, MD (reprint author), Maternal & Child Hlth Bur, US Hlth Resources & Serv Adm, 5600 Fishers Ln,Room 18A-55, Rockville, MD 20857 USA. NR 46 TC 61 Z9 62 U1 1 U2 2 PU AMER PUBLIC HEALTH ASSOC INC PI WASHINGTON PA 1015 FIFTEENTH ST NW, WASHINGTON, DC 20005 USA SN 0090-0036 J9 AM J PUBLIC HEALTH JI Am. J. Public Health PD FEB PY 2000 VL 90 IS 2 BP 245 EP 250 DI 10.2105/AJPH.90.2.245 PG 6 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 278QA UT WOS:000084999400014 PM 10667186 ER PT J AU Redline, S Kapur, VK Sanders, MH Quan, SF Gottlieb, DJ Rapoport, DM Bonekat, WH Smith, PL Kiley, JP Iber, C AF Redline, S Kapur, VK Sanders, MH Quan, SF Gottlieb, DJ Rapoport, DM Bonekat, WH Smith, PL Kiley, JP Iber, C CA Sleep Heart Hlth Res Grp TI Effects of varying approaches for identifying respiratory disturbances on sleep apnea assessment SO AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE LA English DT Article ID MORBIDITY; HYPOPNEA; HEALTH AB Varying approaches to measuring the respiratory disturbance index (RDI) may lead to discrepant estimates of the severity of sleep-disordered breathing (SDB). In this study, we assessed the impact of varying the use of corroborative data (presence and degree of desaturation and/or arousal) to identify hypopneas and apneas. The relationships among 10 RDIs defined by various definitions of apneas and hypopneas were assessed in 5,046 participants in the Sleep Heart Health Study (SHHS) who underwent overnight unattended 12-channel polysomnography (PSC). The magnitude of the median RDI varied 10-fold (i.e., 29.3 when the RDI was based on events identified on the basis of flow or volume amplitude criteria alone to 2.0 for an RDI that required an associated 5% desaturation with events). The correlation between RDIs based on different definitions ranged from 0.99 to 0.68. The highest correlations were among RDIs that required apneas and hypopneas to be associated with some level of desaturation. Lower correlations were observed between RDIs that required desaturation as compared with RDIs defined on the basis of amplitude criteria alone or associated arousal. These data suggest that different approaches for measuring the RDI may contribute to substantial variability in identification and classification of the disorder. C1 Case Western Reserve Univ, Dept Pediat, Cleveland, OH 44106 USA. Univ Washington, Dept Med, Seattle, WA USA. Univ Pittsburgh, Dept Med, Pittsburgh, PA USA. Univ Pittsburgh, Dept Anesthesiol, Pittsburgh, PA USA. Univ Arizona, Dept Med, Tucson, AZ USA. Univ Arizona, Resp Sci Ctr, Tucson, AZ USA. Boston Univ, Dept Med, Boston, MA 02215 USA. NYU, Dept Med, New York, NY 10016 USA. Univ Calif Davis, Dept Med, Davis, CA 95616 USA. Johns Hopkins Univ, Dept Med, Baltimore, MD USA. NHLBI, NIH, Bethesda, MD 20892 USA. Univ Minnesota, Dept Med, Minneapolis, MN 55455 USA. RP Redline, S (reprint author), Rainbow Babies & Childrens Hosp, Div Clin Epidemiol, 11100 Euclid Ave, Cleveland, OH 44106 USA. RI Kapur, Vishesh/K-1054-2014 OI Kapur, Vishesh/0000-0002-5417-1097 FU NHLBI NIH HHS [U01HL53940, U01 HL053938, U01HL53941, UO1HL53938] NR 15 TC 132 Z9 136 U1 0 U2 0 PU AMER LUNG ASSOC PI NEW YORK PA 1740 BROADWAY, NEW YORK, NY 10019 USA SN 1073-449X J9 AM J RESP CRIT CARE JI Am. J. Respir. Crit. Care Med. PD FEB PY 2000 VL 161 IS 2 BP 369 EP 374 PG 6 WC Critical Care Medicine; Respiratory System SC General & Internal Medicine; Respiratory System GA 285ED UT WOS:000085376000009 PM 10673173 ER PT J AU Campo, E Jaffe, ES AF Campo, E Jaffe, ES TI Nodal marginal zone lymphoma - Authors' reply SO AMERICAN JOURNAL OF SURGICAL PATHOLOGY LA English DT Letter ID B-CELL LYMPHOMA C1 Univ Barcelona, Pathol Lab, Hosp Clin, Barcelona, Spain. NCI, Hematopathol Sect, Pathol Lab, NIH, Bethesda, MD 20892 USA. RP Campo, E (reprint author), Univ Barcelona, Pathol Lab, Hosp Clin, Barcelona, Spain. OI Campo, elias/0000-0001-9850-9793 NR 7 TC 1 Z9 1 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0147-5185 J9 AM J SURG PATHOL JI Am. J. Surg. Pathol. PD FEB PY 2000 VL 24 IS 2 BP 315 EP 317 DI 10.1097/00000478-200002000-00035 PG 3 WC Pathology; Surgery SC Pathology; Surgery GA 280ZM UT WOS:000085133700030 ER PT J AU Roper, MH Torres, RSC Goicochea, CGC Andersen, EM Guarda, JSA Calampa, C Hightower, AW Magill, AJ AF Roper, MH Torres, RSC Goicochea, CGC Andersen, EM Guarda, JSA Calampa, C Hightower, AW Magill, AJ TI The epidemiology of malaria in an epidemic area of the Peruvian Amazon SO AMERICAN JOURNAL OF TROPICAL MEDICINE AND HYGIENE LA English DT Article ID PLASMODIUM-FALCIPARUM PARASITEMIA; ANOPHELES-DARLINGI; COMPARATIVE SUSCEPTIBILITY; RONDONIA; BRAZIL; INFECTION; MOSQUITOS; COMMUNITY; VIVAX; TRANSMISSION AB A longitudinal study of malariometric indicators and their association with potential risk factors was conducted during August 1997-July 1998 at Padre Cocha, a village of 1,400 residents in the Peruvian Amazon. The incidence of Plasmodium falciparum infections during the study year was 166/1,000 persons; that of P. vivax was 826/1,000 persons. The mean duration of symptoms prior to diagnosis was 2 days; presenting geometric mean parasite densities were 3,976 parasites/mu l for P. falciparum infections and 2,282 parasites/mu l for P. vivax. There were no malaria-associated deaths. Consistent with the epidemic nature of malaria in the area, the incidence of both parasite species increased with age and there were no age-specific differences in mean parasite densities. No specific occupational risks for malaria were identified. Activities significantly associated with malaria risk reflected local vector behavior and included strolling outdoors after 6:00 PM and arising before 6:00 AM for adults, and attending evening church services for children. C1 USN, Med Res Ctr Detachment, Lima, Peru. Ctr Dis Control & Prevent, Div Parasit Dis, Natl Ctr Infect Dis, Atlanta, GA 30341 USA. Direcc Reg Salud Loreto, Iquitos, Peru. RP Magill, AJ (reprint author), NIAID, Malaria Vaccine Dev Unit, Parasit Dis Lab, NIH, Twinbrook 2,Room 103,12441 Parklawn Dr, Rockville, MD 20852 USA. NR 42 TC 46 Z9 50 U1 0 U2 2 PU AMER SOC TROP MED & HYGIENE PI MCLEAN PA 8000 WESTPARK DRIVE SUITE 130, MCLEAN, VA 22101 USA SN 0002-9637 J9 AM J TROP MED HYG JI Am. J. Trop. Med. Hyg. PD FEB PY 2000 VL 62 IS 2 BP 247 EP 256 PG 10 WC Public, Environmental & Occupational Health; Tropical Medicine SC Public, Environmental & Occupational Health; Tropical Medicine GA 311BN UT WOS:000086864800014 PM 10813480 ER PT J AU Collins, WA Maccoby, EE Steinberg, L Hetherington, EM Bornstein, MH AF Collins, WA Maccoby, EE Steinberg, L Hetherington, EM Bornstein, MH TI Contemporary research on parenting - The case for nature and nurture SO AMERICAN PSYCHOLOGIST LA English DT Review ID MOTHER-CHILD INTERACTION; DEVELOPMENTAL PERSPECTIVE; ENVIRONMENT CORRELATIONS; PSYCHOLOGICAL DISTRESS; ROMANIAN ORPHANAGES; FAMILY ENVIRONMENT; ECONOMIC HARDSHIP; MULTIPLE PATHWAYS; MATERNAL-CARE; MENTAL-HEALTH AB Current findings on parental influences provide more sophisticated and less deterministic explanations than did earlier theory and research on parenting. Contemporary research approaches include (a) behavior-genetic designs, augmented with direct measures of potential environmental influences; (b) studies distinguishing among children with different, genetically influenced predispositions in terms of their responses to different environmental conditions; (c) experimental and quasi-experimental studies of change in children's behavior as a result of their exposure to parents' behavior, after controlling for children's initial characteristics; and (d) research on interactions between parenting and nonfamilial environmental influences and contexts, illustrating contemporary concern with influences beyond the parent-child dyad. These approaches indicate that parental influences on child development are neither as unambiguous as earlier researchers suggested nor as insubstantial as current critics claim. C1 Univ Minnesota, Inst Child Dev, Minneapolis, MN 55455 USA. Stanford Univ, Dept Psychol, Stanford, CA 94305 USA. Temple Univ, Dept Psychol, Philadelphia, PA 19122 USA. Univ Virginia, Dept Psychol, Charlottesville, VA 22903 USA. NICHHD, Bethesda, MD 20892 USA. RP Collins, WA (reprint author), Univ Minnesota, Inst Child Dev, 51 E River Rd, Minneapolis, MN 55455 USA. RI Nguyen, Linh/H-3611-2012 NR 146 TC 633 Z9 657 U1 15 U2 195 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 0003-066X J9 AM PSYCHOL JI Am. Psychol. PD FEB PY 2000 VL 55 IS 2 BP 218 EP 232 DI 10.1037/0003-066X.55.2.218 PG 15 WC Psychology, Multidisciplinary SC Psychology GA 289NP UT WOS:000085629100002 PM 10717969 ER PT J AU Morrell, CH Brant, LJ AF Morrell, CH Brant, LJ TI Lines in random effects plots from the linear mixed-effects model SO AMERICAN STATISTICIAN LA English DT Article DE linear mixed-effects model; plots; random effects ID LONGITUDINAL DATA AB After fitting a linear mixed-effects model to a set of repeated-measures or longitudinal data, it is common practice to plot the estimated random effects. On occasion it may be observed that in these plots a straight line may appear. How did this line arise? What influence does it have on the interpretation of the results from the model? This article demonstrates an artifact that can occur in the plots of random effects. If a cluster has exactly one observation, the plot of any estimated random effect against any other estimated random effect will fall on a straight line. The line in the plot of a pair of estimated random effects may even lie in the opposite direction of the direction suggested by the correlation from the random effects covariance matrix. For clusters with two observations at the same design points of the variables for the random effects, the estimated random effects will lie on a plane. Using an example, we demonstrate the patterns in the plots of the estimated random effects. This illustrates that care must be taken when using plots of the estimated random effects. C1 Loyola Coll, Dept Math Sci, Baltimore, MD 21210 USA. NIA, Gerontol Res Ctr, Baltimore, MD 21224 USA. RP Morrell, CH (reprint author), Loyola Coll, Dept Math Sci, Baltimore, MD 21210 USA. NR 14 TC 3 Z9 3 U1 1 U2 4 PU AMER STATISTICAL ASSOC PI ALEXANDRIA PA 1429 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0003-1305 J9 AM STAT JI Am. Stat. PD FEB PY 2000 VL 54 IS 1 BP 1 EP 4 DI 10.2307/2685602 PG 4 WC Statistics & Probability SC Mathematics GA 333JE UT WOS:000088126500001 ER PT J AU Berger, V AF Berger, V TI Ludbrook,J., and Dudley,H. (1998), "Why permutation tests are superior to t and F tests in biomedical research," The American Statistician, 52, 127-132: Comments by Langbehn, Berger, Higgins and Blair, Mallows, and reply SO AMERICAN STATISTICIAN LA English DT Letter C1 NCI, Bethesda, MD 20892 USA. RP Berger, V (reprint author), NCI, Execut Plaza N,Suite 344,6130 Execut Blvd,MSC 735, Bethesda, MD 20892 USA. NR 6 TC 2 Z9 2 U1 0 U2 3 PU AMER STATISTICAL ASSOC PI ALEXANDRIA PA 1429 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0003-1305 J9 AM STAT JI Am. Stat. PD FEB PY 2000 VL 54 IS 1 BP 85 EP 86 PG 2 WC Statistics & Probability SC Mathematics GA 333JE UT WOS:000088126500016 ER PT J AU Njoku, DB Marchick, MR Graf, ML Pohl, LR AF Njoku, DB Marchick, MR Graf, ML Pohl, LR TI High levels of soluble Fas ligand (sFasL) in the serum of clinical anesthesiologists: A possible role in tolerance to anesthetic hepatitis associated antigens SO ANESTHESIA AND ANALGESIA LA English DT Meeting Abstract C1 Johns Hopkins Med Inst, Baltimore, MD 21205 USA. NHLBI, Mol & Cellular Toxicol Sect, LMI, NIH, Bethesda, MD 20892 USA. NR 1 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0003-2999 J9 ANESTH ANALG JI Anesth. Analg. PD FEB PY 2000 VL 90 IS 2 SU S MA S370 PG 1 WC Anesthesiology SC Anesthesiology GA 279FQ UT WOS:000085034100370 ER PT J AU McDermott, MM Liu, K O'Brien, E Guralnik, JM Criqui, MH Martin, GJ Greenland, P AF McDermott, MM Liu, K O'Brien, E Guralnik, JM Criqui, MH Martin, GJ Greenland, P TI Measuring physical activity in peripheral arterial disease: A comparison of two physical activity questionnaires with an accelerometer SO ANGIOLOGY LA English DT Article ID CALTRAC ACCELEROMETER; CARDIOVASCULAR HEALTH; ENERGY-EXPENDITURE; OLDER ADULTS; EXERCISE; INDEX; ANKLE; RISK; MORTALITY; WALKING AB Peripheral arterial disease (PAD)-related exertional leg pain may limit physical activity, thereby contributing to mobility loss and increasing cardiovascular morbidity and mortality in men and women with PAD. The objectives of this study were: (1) to compare objectively measured physical activity levels between patients with and without PAD, (2) to assess the validity of two physical activity questionnaires in patients with PAD. Twenty PAD patients from a noninvasive vascular laboratory and 21 patients without PAD from a general medicine practice wore an accelerometer continuously for 7 days to measure physical activity objectively. After 7 days, participants completed the leisure time physical activity questionnaire (LTPAQ), derived from the Health Interview Survey, and the Stanford 7-day physical activity recall questionnaire (PARQ). PAD participants had markedly lower physical activity levels than non-PAD participants as measured by accelerometer (803 kcal/week +/-364 (range=284-2,000, median=708) vs 1,750 kcal/week +1,296 (range=882-6,586, median=1,278), p<0.001). For the LTPAQ, physical activity levels in PAD and non-PAD participants were 609 kcal/week +/-576 (range=0-2,085, median=529) vs 832 kcal/week +/-784 (range-53-2,820, median= 623), p=0.128. For the PARQ, physical activity levels in PAD and non-PAD participants were 234 METS/week +/-21 (range=214-301, median=229) vs 238 METS/week +/-11 (range=225-268, median=234), p=0.454, respectively. Pearson's correlation coefficient for the association between the accelerometer and the log-transformed LTPAQ measure was 0.419 (p=0.006). Pearson's correlation coefficient was 0.348 for the association between the accelerometer and the log-transformed PARQ measure of physical activity (p=0.026). In conclusion, PAD patients have significantly lower physical activity levels than non-PAD patients. Two commonly used physical activity questionnaires were less sensitive than objective measurement to the association between PAD and inactivity. C1 Northwestern Univ, Sch Med, Dept Med, Chicago, IL 60611 USA. Northwestern Univ, Sch Med, Dept Prevent Med, Chicago, IL 60611 USA. NIA, Epidemiol Demog & Biometry Program, Bethesda, MD 20892 USA. Univ Calif San Diego, Dept Family & Prevent Med, La Jolla, CA 92093 USA. RP McDermott, MM (reprint author), 675 N St Clair,Suite 18-200, Chicago, IL 60611 USA. FU NCRR NIH HHS [RR-00048]; NHLBI NIH HHS [R01 HL58099] NR 29 TC 61 Z9 61 U1 0 U2 5 PU WESTMINSTER PUBL INC PI GLEN HEAD PA 708 GLEN COVE AVE, GLEN HEAD, NY 11545 USA SN 0003-3197 J9 ANGIOLOGY JI Angiology PD FEB PY 2000 VL 51 IS 2 BP 91 EP 100 PG 10 WC Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 285HF UT WOS:000085383100001 PM 10701716 ER PT J AU DeGraba, T Azhar, S Dignat-George, F Brown, E Boutiere, B Altarescu, G McCarron, R Schiffmann, R AF DeGraba, T Azhar, S Dignat-George, F Brown, E Boutiere, B Altarescu, G McCarron, R Schiffmann, R TI Profile of endothelial and leukocyte activation in Fabry patients SO ANNALS OF NEUROLOGY LA English DT Article ID TUMOR NECROSIS FACTOR; VON-WILLEBRAND-FACTOR; PROCOAGULANT ACTIVITY; REPLACEMENT THERAPY; DISEASE; CELLS; GLYCOSPHINGOLIPIDS; INTERLEUKIN-1; ADHESION; LIPOPROTEINS AB Fabry disease is an X-linked recessive disorder resulting in the deposition of globotriaosylceramide in numerous cell types including vascular endothelial cells. Because this disease is associated with vascular injury and a high recurrence rate of thrombotic events, measurements of factors regulating endothelium and leukocyte interaction may provide insight into the mechanisms leading to a prothrombotic state. Twenty-five patients with Fabry disease and 25 control subjects participated in the study. Plasma from all 25 Fabry patients and 15 of the 25 controls were studied for multiple endothelial factors. Leukocyte integrins were measured by flow cytometry in 21 Fabry patients and 10 controls. The concentrations of soluble intercellular adhesion molecule-1, vascular cell adhesion molecule-1, P-selectin, and plasminogen activator inhibitor were significantly higher and thrombomodulin was significantly lower in Fabry patients. Expression of the integrin CD11b on monocytes was also significantly higher in the Fabry patients. This study reveals measurable evidence for endothelium and leukocyte activation that is consistent with a prothrombotic state in Fabry patients compared with controls. Further investigations of these findings may help to understand the mechanism of stroke in Fabry disease and provide indicators (or markers) of efficacy of future therapeutic intervention. C1 NINDS, Stroke Branch, NIH, Bethesda, MD 20892 USA. NINDS, Dev & Metab Neurol Branch, NIH, Bethesda, MD 20892 USA. USN, Med Res Ctr, Bethesda, MD 20084 USA. Lab Hematol & Immunol, Marseille, France. RP DeGraba, T (reprint author), NINDS, Stroke Branch, NIH, 36 Convent Dr,MSC 4128,Bldg 36,Room 4A-03, Bethesda, MD 20892 USA. NR 30 TC 111 Z9 117 U1 1 U2 3 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0364-5134 J9 ANN NEUROL JI Ann. Neurol. PD FEB PY 2000 VL 47 IS 2 BP 229 EP 233 DI 10.1002/1531-8249(200002)47:2<229::AID-ANA13>3.0.CO;2-T PG 5 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 280TR UT WOS:000085120300013 PM 10665494 ER PT J AU Strunk, T Bubel, S Mascher, B Schlenke, P Kirchner, H Wandinger, KP AF Strunk, T Bubel, S Mascher, B Schlenke, P Kirchner, H Wandinger, KP TI Increased numbers of CCR5(+) interferon-gamma- and tumor necrosis factor-alpha-secreting T lymphocytes in multiple sclerosis patients SO ANNALS OF NEUROLOGY LA English DT Article ID EXACERBATIONS; EXPRESSION; ACTIVATION AB To determine the frequency of in vivo activated T(H)1 lymphocytes; T-cell subsets of 9 multiple sclerosis patients with active disease and 17 healthy controls were analyzed by immunostaining for CCR5, CD26, and their expression of interleukin-2, interferon-gamma, and tumor necrosis factor-alpha. The numbers of CCR5(+) interferon-gamma- and tumor necrosis factor-alpha-producing T cells were significantly increased in the peripheral blood of multiple sclerosis patients. CCR5 expression may be a useful marker to identify effector cells in multiple sclerosis and could be used as a tool for monitoring disease activity. C1 NINDS, Neuroimmunol Branch, NIH, Bethesda, MD 20892 USA. Univ Lubeck, Sch Med, Dept Neurol, D-2400 Lubeck, Germany. Univ Lubeck, Sch Med, Inst Immunol & Transfus Med, D-2400 Lubeck, Germany. RP Wandinger, KP (reprint author), NINDS, Neuroimmunol Branch, NIH, Bldg 10 Room 5B-16,10 Ctr DR MSC 1400, Bethesda, MD 20892 USA. NR 10 TC 63 Z9 65 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0364-5134 J9 ANN NEUROL JI Ann. Neurol. PD FEB PY 2000 VL 47 IS 2 BP 269 EP 273 DI 10.1002/1531-8249(200002)47:2<269::AID-ANA23>3.0.CO;2-G PG 5 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 280TR UT WOS:000085120300023 PM 10665504 ER PT J AU Bielamowicz, S Ludlow, CL AF Bielamowicz, S Ludlow, CL TI Effects of botulinum toxin on pathophysiology in spasmodic dysphonia SO ANNALS OF OTOLOGY RHINOLOGY AND LARYNGOLOGY LA English DT Article DE Botox; botulinum toxin; electromyography; laryngology; larynx; spasmodic dysphonia; voice ID FOCAL LARYNGEAL DYSTONIA; NEUROTOXIN TYPE-A; SPASTIC DYSPHONIA; MUSCLE-ACTIVITY; ABDUCENS MOTONEURONS; INJECTIONS; ADDUCTOR; THERAPY; PATTERN; SPEECH AB To determine the mechanism of symptom relief with treatment by botulinum toxin injection in persons with adductor spasmodic dysphonia (ADSD), we evaluated the effects of unilateral thyroarytenoid muscle injections on both injected and noninjected muscles in 10 subjects with ADSD, using electromyography on bath sides of the larynx before and after treatment. The subjects' speech symptoms were reduced (p = .005) 2 weeks following injection; when the electromyographic study occurred. Muscle activation levels and the numbers of spasmodic muscle bursts decreased significantly (p less than or equal to.03) postinjection in both the injected and noninjected muscles. The reductions in laryngeal muscle bursts correlated with symptom reduction (r greater than or equal to.7) in all muscles. Reductions in laryngeal muscle bursts did not relate to either absolute or normalized levels of muscle activity before or after botulinum toxin injection. The results suggest that changes in the central pathophysiology are responsible for changes in speech symptoms following treatment. C1 NINDS, Laryngeal & Speech Sect, NIH, Bethesda, MD 20892 USA. RP Bielamowicz, S (reprint author), George Washington Univ, Med Ctr, Div Otolaryngol Head & Neck Surg, 2150 Penn Ave NW, Washington, DC 20037 USA. OI Ludlow, Christy/0000-0002-2015-6171 FU NIDCD NIH HHS [Z01 DC 00004] NR 38 TC 42 Z9 42 U1 1 U2 3 PU ANNALS PUBL CO PI ST LOUIS PA 4507 LACLEDE AVE, ST LOUIS, MO 63108 USA SN 0003-4894 J9 ANN OTO RHINOL LARYN JI Ann. Otol. Rhinol. Laryngol. PD FEB PY 2000 VL 109 IS 2 BP 194 EP 203 PG 10 WC Otorhinolaryngology SC Otorhinolaryngology GA 283CD UT WOS:000085257800015 PM 10685573 ER PT J AU Sausville, EA AF Sausville, EA TI Protein kinase antagonists: interim challenges and issues SO ANTI-CANCER DRUG DESIGN LA English DT Article C1 NCI, Div Canc Treatment & Diagnosis, Dev Therapeut Program, Rockville, MD 21852 USA. RP Sausville, EA (reprint author), NCI, Div Canc Treatment & Diagnosis, Dev Therapeut Program, Rockville, MD 21852 USA. NR 6 TC 11 Z9 12 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0266-9536 J9 ANTI-CANCER DRUG DES JI Anti-Cancer Drug Des. PD FEB PY 2000 VL 15 IS 1 BP 1 EP 2 PG 2 WC Biochemistry & Molecular Biology; Oncology; Chemistry, Medicinal; Pharmacology & Pharmacy SC Biochemistry & Molecular Biology; Oncology; Pharmacology & Pharmacy GA 337PG UT WOS:000088370400001 PM 10888031 ER PT J AU Gussio, R Zaharevitz, DW McGrath, CF Pattabiraman, N Kellogg, GE Schultz, C Link, A Kunick, C Leost, M Meijer, L Sausville, EA AF Gussio, R Zaharevitz, DW McGrath, CF Pattabiraman, N Kellogg, GE Schultz, C Link, A Kunick, C Leost, M Meijer, L Sausville, EA TI Structure-based design modifications of the paullone molecular scaffold for cyclin-dependent kinase inhibition SO ANTI-CANCER DRUG DESIGN LA English DT Article DE cyclin-dependent kinase inhibitors; 3-D QSAR; paullones; protein homology modeling; quantum mechanical calculations; structure-based drug design ID CRYSTAL-STRUCTURE; PURINE ANALOGS; CANCER CELLS; CDK2; FLAVOPIRIDOL; ROSCOVITINE; LIBRARIES; THERAPY AB A congeneric series of paullones were characterized using a 3-D QSAR with cyclin-dependent kinase 1 (CDK1) inhibition data. A homology model of CDK1-cyclin B was developed from the crystal structure of CDK2-cyclin A, which subsequently served as the basis for the structure-based design, Paullones were docked into the ATP binding site of the CDK1-cylin B models and were optimized with molecular mechanics. Hydropathic analysts of the paullone-CDK1 complexes were performed after the atom types were assigned based on each ligand's electronic properties calculated from quantum mechanics. Hydropathic descriptors formed a significant multiple regression equation that predicts paullone IC50 data. The results indicate that the combination of hydropathic descriptors with molecular mechanics geometries are sufficient to design overt steric and chemical complementarity of the ligands, However, the electronic properties derived from quantum mechanics helped direct synthetic chemistry efforts to produce ligands that promote butter charge transfer and strengthen hydrogen bonding as facilitated by resonance stabilization, Compounds with low affinity for CDK1,were poor charge accepters and made less than ideal hydrogen bonding arrangements with the receptor. These considerations led to the prediction that structures such as 9-cyanopaullone would be considerably more potent than the parent compound, a finding supported by enzyme inhibition data. Also, 9-nitropaullone emerged as a paullone which also had similar potency in enzyme inhibition as well as a favorable anti-proliferative activity profile in living cells. C1 NCI, Div Canc Treatment & Diagnosis, Dev Therapeut Program, Frederick, MD 21702 USA. NCI, Adv Biomed Comp Ctr, FCRDC, SAIC, Frederick, MD 21702 USA. Virginia Commonwealth Univ, Dept Med Chem, Sch Pharm, Richmond, VA 23298 USA. Virginia Commonwealth Univ, Inst Struct Biol & Drug Discovery, Sch Pharm, Richmond, VA 23298 USA. Univ Hamburg, Abt Pharmazeut Chem, Inst Pharm, D-20146 Hamburg, Germany. CNRS, Biol Stn, F-29682 Roscoff, France. RP Gussio, R (reprint author), NCI, Div Canc Treatment & Diagnosis, Dev Therapeut Program, Frederick, MD 21702 USA. RI Kellogg, Glen/A-8008-2011; Link, Andreas/H-5067-2013 OI Link, Andreas/0000-0003-1262-6636 NR 37 TC 64 Z9 68 U1 0 U2 4 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0266-9536 J9 ANTI-CANCER DRUG DES JI Anti-Cancer Drug Des. PD FEB PY 2000 VL 15 IS 1 BP 53 EP 66 PG 14 WC Biochemistry & Molecular Biology; Oncology; Chemistry, Medicinal; Pharmacology & Pharmacy SC Biochemistry & Molecular Biology; Oncology; Pharmacology & Pharmacy GA 337PG UT WOS:000088370400006 PM 10888036 ER PT J AU Ramaswamy, SV Amin, AG Goksel, S Stager, CE Dou, SJ El Sahly, H Moghazeh, SL Kreiswirth, BN Musser, JM AF Ramaswamy, SV Amin, AG Goksel, S Stager, CE Dou, SJ El Sahly, H Moghazeh, SL Kreiswirth, BN Musser, JM TI Molecular genetic analysis of nucleotide polymorphisms associated with ethambutol resistance in human isolates of Mycobacterium tuberculosis SO ANTIMICROBIAL AGENTS AND CHEMOTHERAPY LA English DT Article ID CELL-WALL; ARABINAN BIOSYNTHESIS; INHIBITION; ARABINOGALACTAN; SMEGMATIS; IDENTIFICATION; COMPLEX; AVIUM; EMBB; DRUG AB Ethambutol (EMB) is a central component of drug regimens used worldwide for the treatment of tuberculosis. To gain insight into the molecular genetic basis of EMB resistance, approximately 2 Mb of five chromosomal regions with 12 genes in 75 epidemiologically unassociated EMB-resistant and 33 EMB-susceptible Mycobacterium tuberculosis strains isolated from human patients were sequenced. Seventy-six: percent of EMB-resistant organisms had an amino acid replacement or other molecular change not found in EMB-susceptible strains. Thirty-eight (51%) EMB-resistant isolates had a resistance-associated mutation in only 1 of the 12 genes sequenced. Nineteen EMB-resistant isolates had resistance-associated nucleotide changes that conferred amino acid replacements or upstream potential regulatory region mutations in two or more genes. Most isolates (68%) with resistance-associated mutations in a single gene had nucleotide changes In embB, a gene encoding an arabinosyltransferase involved in cell wall biosynthesis. The majority of these mutations resulted in amino acid replacements at position 306 or 406 of EmbB, Resistance-associated mutations were also identified in several genes recently shown to be upregulated in response to exposure of M. tuberculosis to EMB in vitro, including genes in the iniA operon, Approximately one-fourth of the organisms studied lacked mutations inferred to participate in EMB resistance, a result indicating that one or more genes that mediate resistance to this drug remain to be discovered. Taken together, the results indicate that there are multiple molecular pathways to the EMB resistance phenotype. C1 NIAID, Rocky Mt Labs, Lab Human Bacterial Pathogenesis, NIH, Hamilton, MT 59840 USA. Baylor Coll Med, Inst Study Human Bacterial Pathogenesis, Dept Pathol, Houston, TX 77030 USA. Publ Hlth Res Inst City New York Inc, TB Ctr, New York, NY 10016 USA. RP Musser, JM (reprint author), NIAID, Rocky Mt Labs, Lab Human Bacterial Pathogenesis, NIH, 903 S 4th St, Hamilton, MT 59840 USA. EM jmusser@niaid.nih.gov FU NIAID NIH HHS [AI-37004] NR 39 TC 152 Z9 171 U1 0 U2 2 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0066-4804 EI 1098-6596 J9 ANTIMICROB AGENTS CH JI Antimicrob. Agents Chemother. PD FEB PY 2000 VL 44 IS 2 BP 326 EP 336 DI 10.1128/AAC.44.2.326-336.2000 PG 11 WC Microbiology; Pharmacology & Pharmacy SC Microbiology; Pharmacology & Pharmacy GA 276WZ UT WOS:000084902500017 PM 10639358 ER PT J AU Groll, AH Mickiene, D Piscitelli, SC Walsh, TJ AF Groll, AH Mickiene, D Piscitelli, SC Walsh, TJ TI Distribution of lipid formulations of amphotericin B into bone marrow and fat tissue in rabbits SO ANTIMICROBIAL AGENTS AND CHEMOTHERAPY LA English DT Article ID PULMONARY ASPERGILLOSIS; LIPOSOMAL FORMULATION; COLLOIDAL DISPERSION; PHARMACOKINETICS; GENTAMICIN; LEISHMANIASIS; PHARMACOLOGY; AMBISOME; SERUM AB The distribution of the three currently available lipid formulations of amphotericin B (AmB) into bone marrow and fat tissue was evaluated in noninfected rabbits. Groups of four animals each received either 1 mg of AmB deoxycholate (D-AmB) per kg of body weight per day or 5 mg of AmB colloidal dispersion, AmB lipid complex, or Liposomal AmB per kg per day for seven doses. Plasma, bone marrow, fat, and liver were collected at autopsy, and AmB concentrations were determined by high-performance liquid chromatography. At the investigated dosages of 5 mg/kg/day, all AmB lipid formulations achieved at least fourfold-higher concentrations in bone marrow than did standard D-AmB at a dosage of 1 mg/kg/day, Concentrations in bone marrow were 62 to 76% of concurrent AmB concentrations in the liver. In contrast, all AmB formulations accumulated comparatively poorly in fat tissue. The results of this study show that high concentrations of AmB can be achieved in the bone marrow after administration of lipid formulations, suggesting their particular usefulness against disseminated fungal infections involving the bone marrow and against visceral leishmaniasis. C1 NCI, Pediat Oncol Branch, Immunocompromised Host Sect, NIH, Bethesda, MD 20892 USA. NIH, Warren G Magnuson Clin Ctr, Dept Pharm, Pharmacokinet Res Lab, Bethesda, MD 20892 USA. RP Walsh, TJ (reprint author), NCI, Pediat Oncol Branch, Immunocompromised Host Sect, NIH, Bldg 10,Room 13N240,10 Ctr Dr, Bethesda, MD 20892 USA. NR 22 TC 11 Z9 14 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 USA SN 0066-4804 J9 ANTIMICROB AGENTS CH JI Antimicrob. Agents Chemother. PD FEB PY 2000 VL 44 IS 2 BP 408 EP 410 DI 10.1128/AAC.44.2.408-410.2000 PG 3 WC Microbiology; Pharmacology & Pharmacy SC Microbiology; Pharmacology & Pharmacy GA 276WZ UT WOS:000084902500030 PM 10639371 ER PT J AU Srinivas, RV AF Srinivas, RV TI May the drug transporter P glycoprotein affect the antiviral activity of human immunodeficiency virus type 1 proteinase inhibitors? Author's reply SO ANTIMICROBIAL AGENTS AND CHEMOTHERAPY LA English DT Letter ID CELL-LINES; EXPRESSION; RESISTANCE C1 NIH, Ctr Sci Review, Bethesda, MD 20892 USA. RP Srinivas, RV (reprint author), NIH, Ctr Sci Review, 6701 Rockledge Dr, Bethesda, MD 20892 USA. NR 8 TC 0 Z9 0 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 USA SN 0066-4804 J9 ANTIMICROB AGENTS CH JI Antimicrob. Agents Chemother. PD FEB PY 2000 VL 44 IS 2 BP 474 EP 474 PG 1 WC Microbiology; Pharmacology & Pharmacy SC Microbiology; Pharmacology & Pharmacy GA 276WZ UT WOS:000084902500050 ER PT J AU Geselowitz, DA Cramer, H Wondrak, EM Player, MR Torrence, PF AF Geselowitz, DA Cramer, H Wondrak, EM Player, MR Torrence, PF TI Fluorescence resonance energy transfer analysis of RNase L-catalyzed oligonucleotide cleavage SO ANTISENSE & NUCLEIC ACID DRUG DEVELOPMENT LA English DT Article ID INTERFERON-TREATED CELLS; DOUBLE-STRANDED-RNA; FLUOROGENIC SUBSTRATE; ANTISENSE CHIMERAS; PROTEIN-SYNTHESIS; 2-5A SYSTEM; ACTIVATION; 2',5'-OLIGOADENYLATE; DEGRADATION; BINDING AB A method is described for monitoring the cleavage of an oligoribonucleotide substrate by the 2-5A-dependent RNase L based on fluorescence resonance energy transfer (FRET). The oligoribonucleotide, rC(11)U(2)C(7), was labeled covalently at its 5'-terminus with fluorescein and at its 3'-terminus with rhodamine to provide a substrate for RNase L. On cleavage, the fluorescence at 538 mm (with 485 nm excitation) increased by a factor of 2.8, allowing real-time quantitation of the reaction progress. The method was performed easily in a 96-well plate format and allowed quantitative high throughput analyses of RNase L activity with different activators. C1 NIDDKD, Sect Biomed Chem, Med Chem Lab, NIH, Bethesda, MD 20892 USA. RP Torrence, PF (reprint author), No Arizona Univ, Dept Chem, Bldg 020, Flagstaff, AZ 86011 USA. NR 32 TC 7 Z9 7 U1 1 U2 5 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1087-2906 J9 ANTISENSE NUCLEIC A JI Antisense Nucleic Acid Drug Dev. PD FEB PY 2000 VL 10 IS 1 BP 45 EP 51 DI 10.1089/oli.1.2000.10.45 PG 7 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Research & Experimental Medicine GA 289VD UT WOS:000085641900006 PM 10726660 ER PT J AU Bray, M Driscoll, J Huggins, JW AF Bray, M Driscoll, J Huggins, JW TI Treatment of lethal Ebola virus infection in mice with a single dose of an S-adenosyl-L-homocysteine hydrolase inhibitor SO ANTIVIRAL RESEARCH LA English DT Article DE filovirus; Ebola virus; antiviral therapy; S-adenosyl-L-homocysteine hydrolase ID SPECTRUM ANTIVIRAL ACTIVITY; ADENOSYLHOMOCYSTEINE HYDROLASE; CARBOCYCLIC 3-DEAZAADENOSINE; 3-DEAZANEPLANOCIN-A; AGENT; ANALOG; PHARMACOKINETICS; PROPHYLAXIS; ACTIVATION; THERAPY AB Ebola Zaire virus causes lethal hemorrhagic fever in humans, for which there is no effective treatment. A variety of adenosine analogues inhibit the replication of Ebola virus in vitro, probably by blocking the cellular enzyme, S-adenosyl-L-homocysteine hydrolase, thereby indirectly limiting methylation of the 5' cap of viral messenger RNA. We previously observed that adult, immunocompetent mice treated thrice daily for 9 days with 2.2-20 mg/kg of an adenosine analogue, carbocyclic 3-deazaadenosine, were protected against lethal Ebola virus challenge. We now report that a single inoculation of 80 mg/kg or less of the same substance, or of 1 mg/kg or less of another analogue, 3-deazaneplanocin A, provides equal or better protection, without causing acute toxicity. One dose of drug given on the first or second day after virus infection reduced peak viremia more than 1000-fold, compared with mock-treated controls, and resulted in survival of most or all animals. Therapy was less effective when administered on the day of challenge, or on the third day postinfection. Single or multiple doses of the same medications suppressed Ebola replication in severe combined immunodeficient mice, but even daily treatment for 15 consecutive days did not eliminate the infection. Published by Elsevier Science B.V. C1 USA, Med Res Inst Infect Dis, Dept Viral Therapeut, Div Virol, Ft Detrick, MD 21702 USA. NCI, Med Chem Lab, Bethesda, MD 20892 USA. RP Bray, M (reprint author), USA, Med Res Inst Infect Dis, Dept Viral Therapeut, Div Virol, 1425 Porter St, Ft Detrick, MD 21702 USA. NR 39 TC 86 Z9 94 U1 2 U2 16 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0166-3542 J9 ANTIVIR RES JI Antiviral Res. PD FEB PY 2000 VL 45 IS 2 BP 135 EP 147 DI 10.1016/S0166-3542(00)00066-8 PG 13 WC Pharmacology & Pharmacy; Virology SC Pharmacology & Pharmacy; Virology GA 301VK UT WOS:000086330800006 PM 10809022 ER PT J AU Yu, HN Tyrrell, D Cashel, J Guo, NH Vogel, T Sipes, JM Lam, L Fillit, HM Hartman, J Mendelovitz, S Panel, A Roberts, DD AF Yu, HN Tyrrell, D Cashel, J Guo, NH Vogel, T Sipes, JM Lam, L Fillit, HM Hartman, J Mendelovitz, S Panel, A Roberts, DD TI Specificities of heparin-binding sites from the amino-terminus and type 1 repeats of thrombospondin-1 SO ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS LA English DT Article DE heparin-binding proteins; thrombospondins; peptides; affinity chromatography; oligosaccharides ID FIBROBLAST-GROWTH-FACTOR; VASCULAR ENDOTHELIAL-CELLS; HUMAN-MELANOMA CELLS; I REPEATS; PLATELET THROMBOSPONDIN; SULFATE PROTEOGLYCAN; CORE PROTEIN; TUMOR-GROWTH; PEPTIDES; EXPRESSION AB Interactions of heparin with intact human thrombospondin-l (TSP1) and with two heparin-binding fragments of TSP1 were characterized using chemically modified heparins, a vascular heparan sulfate proteoglycan, and a series of heparin oligosaccharides prepared by partial deaminative cleavage. The avidity of TSP1 binding increased with oligosaccharide size, with plateaus at 4 to 6 and at 8 to 10 monosaccharide units, The dependence on oligosaccharide size for binding to the recombinant amino-terminal heparin-binding domain of TSP1 was the same as that of the intact TSP1 molecule but differed from that of a synthetic heparin-binding peptide from the type 1 repeats, suggesting that the interaction between intact TSP1 and heparin is primarily mediated by the aminoterminal domain. Based on activities of chemically modified heparins, binding to TSP1 depended primarily on 2-N- and 6-O-sulfation of glucosamine and to a lesser degree on 2,3-O-sulfation and the carboxyl residues of the uronic acids. In contrast, all of these modifications were required for binding of heparin to the type 1 repeat peptides. Affinity purification of heparin octasaccharides on immobilized TSP1 type 1 repeat peptides revealed a preference for oligosaccharides containing the disaccharide sequence IdoA(2-OSO3)alpha 1-4-GlcNS(6-OSO3). Binding of these oligosaccharides to the peptide required the Trp residues. These data demonstrate that the heparin-binding specificities of intact TSP1 and peptides from the type 1 repeats overlap with that of basic fibroblast growth factor (FGF2) and are consistent with the ability of these TSP1-derived molecules to inhibit FGF2-stimulated angiogenesis. (C) 2000 Academic Press. C1 NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. Glycomed Inc, Alameda, CA 94501 USA. BioTechnol Gen Ltd, Rehovot, Israel. Inst Study Aging, New York, NY 10153 USA. CUNY, Mt Sinai Med Ctr, Dept Geriatr, New York, NY 10029 USA. RP Roberts, DD (reprint author), NCI, Pathol Lab, NIH, Bldg 10,Room 2A33,10 Ctr Dr,MSC 1500, Bethesda, MD 20892 USA. RI Roberts, David/A-9699-2008 OI Roberts, David/0000-0002-2481-2981 NR 70 TC 20 Z9 22 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0003-9861 J9 ARCH BIOCHEM BIOPHYS JI Arch. Biochem. Biophys. PD FEB 1 PY 2000 VL 374 IS 1 BP 13 EP 23 DI 10.1006/abbi.1999.1597 PG 11 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 278PX UT WOS:000084999100003 PM 10640391 ER PT J AU Jonkman, MF Schuur, J Dijk, F Heeres, K de Jong, MCJM van der Meer, JB Yancey, KB Pas, HH AF Jonkman, MF Schuur, J Dijk, F Heeres, K de Jong, MCJM van der Meer, JB Yancey, KB Pas, HH TI Inflammatory variant of epidermolysis bullosa acquisita with IgG autoantibodies against type VII collagen and laminin alpha 3 SO ARCHIVES OF DERMATOLOGY LA English DT Article; Proceedings Paper CT Joint Society Meeting of the European-Academy-of-Dermatology-and-Venereology/Groupe Immunodermatologie, Societe-Francaise-de-Dermatologie CY OCT 07, 1998 CL NICE, FRANCE SP European Acad Dermatol & Venereol, Soc Francaise Dermatol,Grp ImmunoDermatol ID BASEMENT-MEMBRANE; INDIRECT IMMUNOFLUORESCENCE; AUTO-ANTIGEN; SKIN; AUTOIMMUNE; ANTIBODIES; EPILIGRIN; DISEASE; CHAIN; IDENTIFICATION AB Background: The inflammatory variant of epidermolysis bullosa acquisita (EBA) may clinically closely resemble bullous or cicatricial pemphigoid. Patients with inflammatory EBA have IgG autoantibodies against type VII collagen. Patients with anti-epiligrin cicatricial pemphigoid have IgG autoantibodies against laminin 5. Observation: We describe a patient with inflammatory EBA exhibiting nonscarring oral and vaginal involvement. Indirect immunofluorescence using skin substrate lacking an epidermal basement membrane molecule, direct immunoelectron microscopy, immuno-blot, and immunoprecipitation studies revealed the simultaneous presence of circulating IgG autoantibodies against type VII collagen and laminin alpha 3. A final diagnosis of EBA was based on the sublamina densa level of blister formation. Conclusion: This case illustrates the clinical and immunological overlap between EBA and anti-epiligrin cicatricial pemphigoid, a unique finding that may have developed as a consequence of epitope spreading. C1 Univ Groningen, Dept Dermatol, Groningen, Netherlands. Univ Groningen, Lab Cell Biol & Electron Microscopy, Groningen, Netherlands. Natl Career Inst, Dermatol Branch, NIH, Bethesda, MD USA. RP Jonkman, MF (reprint author), Univ Groningen Hosp, Dept Dermatol, Hanzepl 1, NL-9713 E2 Groningen, Netherlands. RI Jonkman, Marcel/H-4647-2011; Pas, Hendri/G-9749-2011 OI Pas, Hendri/0000-0001-8823-2591 NR 34 TC 35 Z9 40 U1 0 U2 1 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0003-987X J9 ARCH DERMATOL JI Arch. Dermatol. PD FEB PY 2000 VL 136 IS 2 BP 227 EP 231 DI 10.1001/archderm.136.2.227 PG 5 WC Dermatology SC Dermatology GA 283CK UT WOS:000085258400010 PM 10677099 ER PT J AU Siegel, KL Metman, LV AF Siegel, KL Metman, LV TI Effects of bilateral posteroventral pallidotomy on gait of subjects with Parkinson disease SO ARCHIVES OF NEUROLOGY LA English DT Article ID MEDIAL PALLIDOTOMY; GPI PALLIDOTOMY; STRIDE LENGTH; DETERMINANTS; HYPOKINESIA; STABILITY; EFFICACY; THERAPY; DOPA AB Background: Most studies documenting the effect of pallidotomy on parkinsonian gait have reported unilateral surgery and used qualitative scales or timed tests that only provide measures of walking speed. Objective: To document the effect of bilateral posteroventral pallidotomy on the walking patterns of patients with Parkinson disease (PD). Design: Case series of gait evaluations performed 1 month before and 1 month after surgery, with antiparkinson medication withheld for 8 hours overnight. Setting: Movement analysis laboratory of a clinical research center. Patients: Consecutive sample of 8 men and 3 women with a diagnosis of PD scheduled for bilateral pallidotomy. Intervention: Bilateral posteroventral pallidotomy. Main Outcome Measures: A 3-dimensional motion-capture system allowed calculation of temporal and spatial measurements and joint angular displacements of the lower extremities and trunk during gait. Results: Pallidotomy significantly increased average walking speed from 0.214 statures/s preoperatively to 0.440 statures/s postoperatively (where stature indicates body height) (P = .03). ill faster postoperative walking speed was achieved almost exclusively by increasing average stride length from 0.24 to 0.47 statures (P = .03) rather than changing average gait cycle time (1.32 to 1.37 seconds; P = .08). A forward stepwise multiple regression analysis (P<.001) revealed that 96% of the change in stride length postoperatively could be explained by the combination of changes in foot-floor angle, knee, and hip excursion during gait. Conclusions: Bilateral posteroventral pallidotomy was associated with a 2-fold increase in walking speed. Previous studies have demonstrated that walking speed is an important indicator of locomotor performance and level of disability in patients with PD, so the increase in postoperative walking speed likely provided a functional benefit. C1 NINDS, Dept Rehabil Med, Biomech Lab, Warren Grant Magnuson Clin Ctr,NIH, Bethesda, MD 20892 USA. NINDS, Expt Therapeut Branch, NIH, Bethesda, MD 20892 USA. RP Siegel, KL (reprint author), NINDS, Dept Rehabil Med, Biomech Lab, Warren Grant Magnuson Clin Ctr,NIH, Bldg 10,Rm 6S235,10 Ctr Dr,Mail Stop MSC 1604, Bethesda, MD 20892 USA. RI Siegel, Karen Lohmann/B-5898-2008; OI Siegel, Karen Lohmann/0000-0002-0788-6612 NR 36 TC 20 Z9 21 U1 1 U2 1 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0003-9942 J9 ARCH NEUROL-CHICAGO JI Arch. Neurol. PD FEB PY 2000 VL 57 IS 2 BP 198 EP 204 DI 10.1001/archneur.57.2.198 PG 7 WC Clinical Neurology SC Neurosciences & Neurology GA 282GB UT WOS:000085208700005 PM 10681077 ER PT J AU Olanow, CW AF Olanow, CW CA Tasmar Advisory Panel TI Tolcapone and hepatotoxic effects SO ARCHIVES OF NEUROLOGY LA English DT Article ID IMPROVES MOTOR FUNCTION; DOUBLE-BLIND; PARKINSONIAN-PATIENTS; INHIBITOR TOLCAPONE; LEVODOPA; MULTICENTER; TRIAL AB Four patients with Parkinson disease have recently been described in whom severe hepatic dysfunction developed in association with tolcapone therapy. These reports led to the introduction of a "black box" warning and more intensive monitoring requirements in the United States. A review of these cases and all clinical trials indicates that liver dysfunction did not develop in any patient who had received monitoring of liver function according to the original prescribing information. Virtually all instances of liver enzyme abnormality and clinical liver dysfunction occurred within 6 months of initiating treatment, To assess the current role of tolcapone therapy in Parkinson disease, a panel of neurologists and hepatologists was convened. Consensus was reached with respect to the following: (1) Tolcapone is an effective agent in the treatment of patients with fluctuating Parkinson disease. (2) The risk of developing irreversible liver injury is negligible with appropriate monitoring, (3) It may be possible to reduce the frequency of monitoring after 6 months of treatment. (4) The requirement that tolcapone be withdrawn if liver enzymes are elevated above the upper limit of normal on a single occasion is unnecessarily restrictive. It was concluded that tolcapone, when used as an adjunct to levodopa, is an effective anti-parkinsonian agent and that less frequent monitoring after 6 months, with an action limit of 2 to 3 times the upper limit of normal, is sufficient to ensure safety in patients who are deriving benefit from the drug. C1 Mt Sinai Sch Med, Dept Neurol, New York, NY 10029 USA. NIH, Bethesda, MD 20892 USA. Albany Med Coll, Albany, NY 12208 USA. Columbia Univ, Coll Med, New York, NY 10027 USA. Rush Med Coll, Chicago, IL 60612 USA. Univ S Florida, Coll Med, Tampa, FL 33612 USA. Ohio State Univ, Sch Med, Columbus, OH 43210 USA. Univ Kansas, Sch Med, Kansas City, KS 66103 USA. Wayne State Univ, Sch Med, Detroit, MI 48202 USA. Boston Univ, Sch Med, Boston, MA 02118 USA. Univ Texas, Hlth Sci Ctr, San Antonio, TX 78284 USA. Univ So Calif, Sch Med, Los Angeles, CA 90089 USA. Univ Michigan, Med Ctr, Ann Arbor, MI 48109 USA. Univ Miami, Sch Med, Miami, FL 33136 USA. Armed Forces Inst Pathol, Washington, DC 20306 USA. RP Olanow, CW (reprint author), Mt Sinai Sch Med, Dept Neurol, 1 Gustave Levy Pl,Box 1137, New York, NY 10029 USA. NR 11 TC 93 Z9 94 U1 0 U2 3 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0003-9942 J9 ARCH NEUROL-CHICAGO JI Arch. Neurol. PD FEB PY 2000 VL 57 IS 2 BP 263 EP 267 DI 10.1001/archneur.57.2.263 PG 5 WC Clinical Neurology SC Neurosciences & Neurology GA 282GB UT WOS:000085208700016 PM 10681087 ER PT J AU Hortin, GL King, C Miller, KD Kopp, JB AF Hortin, GL King, C Miller, KD Kopp, JB TI Detection of indinavir crystals in urine - Dependence on method of analysis SO ARCHIVES OF PATHOLOGY & LABORATORY MEDICINE LA English DT Article ID ACUTE-RENAL-FAILURE; PROTEASE INHIBITOR; URETERAL OBSTRUCTION; NEPHROLITHIASIS; UROLITHIASIS; HIV; MICROSCOPY; PERFORMANCE; L-735,524; SECONDARY AB Objectives.-To determine the frequency of crystalluria in patients treated with the human immunodeficiency virus protease inhibitor indinavir and to compare methods of detecting crystalluria. Methods.-A total of 308 freshly voided urine specimens from 168 patients treated with indinavir were evaluated by manual microscopy of sediment and microscopy with an automated workstation and by dipstick analysis. Results.-Crystals were detected in 22%, 31%, or 32% of specimens using, respectively, an automated workstation, manual microscopy, or both methods. Proteinuria or hemoglobinuria occurred significantly more often in specimens with (28%) than without (18%) crystals. Frequency of crystalluria was unrelated to specific gravity, but it increased at higher pH. Crystals were detected in 21% of specimens with pH less than 6 and 42% of specimens with pH of 6 or higher. Conclusions.-Crystalluria occurs in more than 30% of urine specimens from patients treated with indinavir, but detection rates vary substantially with method of analysis. Manual microscopy detected crystalluria 41% more often than did an automated workstation. C1 NIDDKD, Dept Clin Pathol, NIH, Bethesda, MD 20892 USA. NIDDKD, Crit Care Med Dept, NIH, Bethesda, MD 20892 USA. NIDDKD, Warren Grant Magnuson Clin Ctr, NIH, Bethesda, MD 20892 USA. NIDDKD, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. RP Hortin, GL (reprint author), NIDDKD, Dept Clin Pathol, NIH, Bldg 10,Room 2C-407,10 Ctr Dr, Bethesda, MD 20892 USA. OI Kopp, Jeffrey/0000-0001-9052-186X NR 39 TC 12 Z9 14 U1 0 U2 0 PU COLLEGE AMER PATHOLOGISTS PI NORTHFIELD PA C/O KIMBERLY GACKI, 325 WAUKEGAN RD, NORTHFIELD, IL 60093-2750 USA SN 0003-9985 J9 ARCH PATHOL LAB MED JI Arch. Pathol. Lab. Med. PD FEB PY 2000 VL 124 IS 2 BP 246 EP 250 PG 5 WC Medical Laboratory Technology; Medicine, Research & Experimental; Pathology SC Medical Laboratory Technology; Research & Experimental Medicine; Pathology GA 283VQ UT WOS:000085297100013 PM 10656734 ER PT J AU Matsui, K Takeda, K Yu, ZX Travis, WD Moss, J Ferrans, VJ AF Matsui, K Takeda, K Yu, ZX Travis, WD Moss, J Ferrans, VJ TI Role for activation of matrix metalloproteinases in the pathogenesis of pulmonary lymphangioleiomyomatosis SO ARCHIVES OF PATHOLOGY & LABORATORY MEDICINE LA English DT Article ID UTERINE CERVICAL FIBROBLASTS; HORMONAL-REGULATION; TISSUE INHIBITOR; BREAST-CARCINOMA; CELL-SURFACE; GELATINASE-B; EXPRESSION; PROGESTERONE; MMP-2; MT1-MMP AB Background.-Matrix metalloproteinases (MMPs) have been shown to be involved in the pathogenesis of the destructive pulmonary lesions in patients with lymphangioleiomyomatosis (LAM); in the present report, the activation of these enzymes is examined. Objective.-To evaluate the role of MMPs and their activating enzymes, immunohistochemical and confocal microscopic techniques were used to localize alpha-smooth muscle actin (alpha-SMA), HMB-45, proliferating cell nuclear antigen (PCNA), MMP-2, membrane-type 1 MMP (MT1-MMP), MT2-MMP, and MT3-MMP in lung tissues from 10 women with LAM. Tissue samples were obtained from 5 patients before treatment and in 5 patients after hormone treatment (progesterone and/or tamoxifen citrate). Results.-Staining for alpha-SMA and MMP-2 was present in all the abnormal smooth muscle cells (LAM cells) in both groups. The percentages of PCNA-, MMP-2-, or MT1-MMP-positive LAM cells were much higher in the untreated group than in the treated group, whereas the percentages of HMB-45-reactive LAM cells were similar in both groups. The reactions for MT1-MMP and PCNA were preferentially localized in small spindle-shaped LAM cells; the reaction for HMB-45 was found in large epithelioid LAM cells. Many of the PCNA-positive cells were also positive for MT1-MMP. Staining for MT2-MMP and MT3-MMP was negative. Conclusions.-This study demonstrates an association between cellular proliferation and the presence of MT1-MMP in LAM cells. The activation of MMP-2 by MT1-MMP may play an important role in the destruction of lung tissue in this disorder. C1 NHLBI, Pathol Sect, NIH, Bethesda, MD 20892 USA. NHLBI, Pulm Crit Care Med Branch, NIH, Bethesda, MD 20892 USA. Armed Forces Inst Pathol, Washington, DC 20306 USA. RP Ferrans, VJ (reprint author), NHLBI, Pathol Sect, NIH, Bldg 10,Room 2N240,10 Ctr Dr,MSC 1518, Bethesda, MD 20892 USA. NR 37 TC 87 Z9 92 U1 0 U2 0 PU COLLEGE AMER PATHOLOGISTS PI NORTHFIELD PA C/O KIMBERLY GACKI, 325 WAUKEGAN RD, NORTHFIELD, IL 60093-2750 USA SN 0003-9985 J9 ARCH PATHOL LAB MED JI Arch. Pathol. Lab. Med. PD FEB PY 2000 VL 124 IS 2 BP 267 EP 275 PG 9 WC Medical Laboratory Technology; Medicine, Research & Experimental; Pathology SC Medical Laboratory Technology; Research & Experimental Medicine; Pathology GA 283VQ UT WOS:000085297100016 PM 10656737 ER PT J AU Klein-Soyer, C Azorsa, DO Cazenave, JP Lanza, F AF Klein-Soyer, C Azorsa, DO Cazenave, JP Lanza, F TI CD9 participates in endothelial cell migration during in vitro wound repair SO ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY LA English DT Article; Proceedings Paper CT XVIth Congress of the International-Society-on-Thrombosis-and-Haemostasis CY JUN 06-12, 1997 CL FLORENCE, ITALY SP Int Soc Thrombosis & Haemostasis DE CD9; integrins; endothelial cells; cell migration; wound repair ID FIBROBLAST GROWTH-FACTOR; GAMMA-II RECEPTOR; MONOCLONAL-ANTIBODY; PLATELET ACTIVATION; TYROSINE PHOSPHORYLATION; IN-VITRO; INTEGRIN; ANGIOGENESIS; ANTIGEN; EXPRESSION AB CD9, a widely expressed membrane protein of the tetraspanin family, has been implicated in diverse functions, such as signal transduction, cell adhesion, and cell motility, We tested the effects of an anti-CD9 monoclonal antibody (ALMA.1) on the migration and proliferation of human vascular endothelial cells (ECs) during repair of an in vitro mechanical wound mimicking angiogenic processes. ALMA.1 induced dose-dependent inhibition of wound repair with a 35 +/- 1.5% decrease at 20 mu g/mL. Only cell migration was affected, because the rate of proliferation of ECs at the lesion margin was not modified and because the inhibition of repair was also observed for nonproliferating irradiated ECs. Monoclonal antibodies against CD63 tetraspanin (H5C6) and control mouse IgG (MOPC-21) were inactive. CD9, one of the most abundant proteins at the surface of ECs, colocalized with beta(1) or beta(3) integrins on EC membranes in double-labeling immunofluorescence experiments with ALMA.1 and an anti-beta(1) (4B4) or anti-beta(3) (SDF.3) monoclonal antibody. Moreover, ALMA.1 and 4B4 had additive inhibitory effects on lesion repair, whereas 4B4 alone also inhibited EC proliferation In transmembrane Boyden-type assays, ALMA.1 induced dose-dependent inhibition of EC migration toward fibronectin and vitronectin with 45+/-6% and 31+/-10% inhibition, respectively, at 100 mu g/mL. 4B4 inhibited migration toward fibronectin at 10 mu g/mL but had no effect in the case of vitronectin, Adhesion of ECs to immobilized anti-CD9 monoclonal antibodies induced tyrosine-phosphorylated protein levels similar to those observed during interactions with beta(1) or beta(3) integrins. These results point to the involvement of CD9 in EC adhesion and migration during lesion repair and angiogenesis, probably through cooperation with integrins. As such, CD9 is a potential target to inhibit angiogenesis in metastatic and atherosclerotic processes. C1 Etablissement Transfus Sanguine, INSERM, U311, Strasbourg, France. Natl Human Genome Res Inst, Canc Genet Lab, NIH, Bethesda, MD USA. RP Klein-Soyer, C (reprint author), Etablissement Transfus Sanguine, INSERM, U311, 10 Rue Spielmann, BP 36, Strasbourg, France. RI Lanza, Francois/H-9252-2016 OI Lanza, Francois/0000-0002-5802-4748 NR 62 TC 45 Z9 46 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1079-5642 J9 ARTERIOSCL THROM VAS JI Arterioscler. Thromb. Vasc. Biol. PD FEB PY 2000 VL 20 IS 2 BP 360 EP 369 PG 10 WC Hematology; Peripheral Vascular Disease SC Hematology; Cardiovascular System & Cardiology GA 281UM UT WOS:000085179200012 PM 10669631 ER PT J AU Brousseau, ME Kauffman, RD Herderick, EE Demosky, SJ Evans, W Marcovina, S Santamarina-Fojo, S Brewer, HB Hoeg, JM AF Brousseau, ME Kauffman, RD Herderick, EE Demosky, SJ Evans, W Marcovina, S Santamarina-Fojo, S Brewer, HB Hoeg, JM TI LCAT modulates atherogenic plasma lipoproteins and the extent of atherosclerosis only in the presence of normal LDL receptors in transgenic rabbits SO ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY LA English DT Article DE familial hypercholesterolemia; metabolism; gene therapy; WHHL rabbits; low density lipoproteins ID HIGH-DENSITY-LIPOPROTEIN; LECITHIN-CHOLESTEROL ACYLTRANSFERASE; CORONARY HEART-DISEASE; ESTER TRANSFER PROTEIN; FAMILIAL HYPERCHOLESTEROLEMIA; APOLIPOPROTEIN-E; BLOOD-PLASMA; WHHL RABBIT; METABOLISM; GENE AB Elevated low density lipoprotein cholesterol (LDL-C) and reduced high density lipoprotein cholesterol (HDL-C) concentrations are independent risk factors for coronary heart disease. We have previously demonstrated that overexpression of an enzyme with a well established role in HDL metabolism, lecithin:cholesterol acyltransferase (LCAT), in New Zealand White rabbits not only raises HDL-C concentrations but reduces those of LDL-C as well, ultimately preventing diet-induced atherosclerosis. In the present study, the human LCAT gene (hLCAT) was introduced into LDL receptor (LDLr)-deficient (Watanabe heritable hyperlipidemic) rabbits to (1) investigate the role of the LDLr pathway in the hLCAT-mediated reductions of LDL-C and (2) determine the influence of hLCAT overexpression on atherosclerosis susceptibility in an animal model of familial hypercholesterolemia. Heterozygosity or homozygosity for the LDLr defect was determined by polymerase chain reaction, and 3 groups of hLCAT-transgenic (hLCAT+) rabbits that differed in LDLr status were established: (1) LDLr wild-type (LDLr+/+), (2) LDLr heterozygotes (LDLr+/-), and (3) LDLr homozygotes (LDLr-/--). Data for hLCAT+ rabbits were compared with those of nontransgenic (hLCAT-) rabbits of the same LDLr status. Plasma HDL-C concentrations were significantly elevated in the hLCAT+ animals of each LDLr status. However, LDL-C levels were significantly reduced only in hLCAT+/LDLr+/+ and hLCAT+/LDLr+/- rabbits but not in hLCAT+/LDLr-/- rabbits (405+/-14 versus 392+/-31 mg/dL), Metabolic studies revealed that the fractional catabolic rate (FCR, d(-1)) of LDL apolipoprotein (apo) B-100 was increased in hLCAT+/LDLr+/+ (26+/-4 versus 5+/-0) and hLCAT+/LDLr+/- (4+/-1 versus 1+/-0) rabbits, whereas the FCR of LDL apoB-100 in both groups of LDLr-/-- rabbits was nearly identical (0.16+/-0.02 versus 0.15+/-0.02). Consistently, neither aortic lipid concentrations nor the extent of aortic atherosclerosis was significantly different between hLCAT+/LDLr-/- and hLCAT-/LDLr-/- rabbits. Significant correlations were observed between the percent of aortic atherosclerosis and both LDL-C (r=0.985) and LDL apoB-100 FCR (-0.745), as well as between LDL-C and LDL apoB-100 FCR (-0.866). These data are the first to establish that LCAT modulates LDL metabolism via the LDLr pathway, ultimately influencing atherosclerosis susceptibility. Moreover, LCAT's antiatherogenic effect requires only a single functional LDLr allele, identifying LCAT as an attractive gene therapy candidate for the majority of dyslipoproteinemic patients. C1 NHLBI, Mol Dis Branch, NIH, Bethesda, MD 20892 USA. Ohio State Univ, Ctr Biomed Engn, Columbus, OH 43210 USA. Univ Washington, Sch Med, NW Lipid Res Labs, Seattle, WA USA. RP Brousseau, ME (reprint author), Tufts Univ, Human Nutr Res Ctr Aging, USDA, JM,Lipid Metab Lab, 711 Washington St, Boston, MA 02111 USA. NR 35 TC 34 Z9 42 U1 1 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1079-5642 J9 ARTERIOSCL THROM VAS JI Arterioscler. Thromb. Vasc. Biol. PD FEB PY 2000 VL 20 IS 2 BP 450 EP 458 PG 9 WC Hematology; Peripheral Vascular Disease SC Hematology; Cardiovascular System & Cardiology GA 281UM UT WOS:000085179200024 PM 10669643 ER PT J AU Feng, DL Tofler, GH Larson, MG O'Donnell, CJ Lipinska, I Schmitz, C Sutherland, PA Johnstone, MT Muller, JE D'Agostino, RB Levy, D Lindpaintner, K AF Feng, DL Tofler, GH Larson, MG O'Donnell, CJ Lipinska, I Schmitz, C Sutherland, PA Johnstone, MT Muller, JE D'Agostino, RB Levy, D Lindpaintner, K TI Factor VII gene polymorphism, factor VII levels, and prevalent cardiovascular disease - The Framingham Heart Study SO ARTERIOSCLEROSIS THROMBOSIS AND VASCULAR BIOLOGY LA English DT Article DE factor VII; genetics; polymorphisms; cardiovascular disease; risk factors ID COAGULATION-FACTOR-VII; PLASMA FACTOR-VII; ACUTE CORONARY SYNDROMES; DIETARY-FAT INTAKE; MYOCARDIAL-INFARCTION; UNSTABLE ANGINA; TISSUE FACTOR; HEMOSTATIC FUNCTION; BLOOD-COAGULATION; ANTIGEN LEVELS AB Elevated factor VII levels have been associated with increased cardiovascular risk in some studies. The arginine/glutamine (Arg/Gln) polymorphism of the factor VII gene has been previously shown to modify factor VII levels. However, the presence of a gene/environment interaction on factor VII levels or a link with cardiovascular disease (CVD) remains uncertain, We studied subjects from the Framingham Heart Study to determine (1) the extent to which this genetic polymorphism affects factor VII levels; (2) whether interactions exist between this polymorphism and environmental factors on factor VII levels; and (3) the association between the polymorphism and CVD, Genotype data and factor VII antigen levels were available in 1816 subjects. Factor VII levels differed significantly among genotypes in an additive fashion: Gin homozygous, 82.7+/-2.5%; heterozygous, 92.2+/-0.7%; and Arg homozygous, 100.5+/-0.4% (P<0.0001). The polymorphism was the strongest, single predictor of factor VII levels, explaining 7.7% of the total variance of factor VII levels, whereas other traditional risk factors combined explained an additional 11.5% of the variance. There was an interaction (P=0.02) between the genotype and total cholesterol on factor VII levels, such that the correlation coefficient and slope (factor VII level/total cholesterol) were greatest in Gln/Gln subjects. Among 3204 subjects characterized for genotype and CVD, there was no significant relationship between the genotype and CVD (P=0.12), In the Framingham Heart Study, the Arg/Gln polymorphism was significantly associated with factor VII antigen levels. The strength of the association suggests that generic variation plays an important role in determining factor VII levels. However, despite being associated with factor VII levels, the Arg/Gln polymorphism was not associated with prevalent CVD. C1 Harvard Univ, Sch Med, Brigham & Womens Hosp, Div Cardiovasc, Boston, MA 02115 USA. Harvard Univ, Sch Med, Beth Israel Deaconess Med Ctr, Inst Prevent Cardiovasc Dis, Boston, MA USA. Royal N Shore Hosp, Sydney, NSW, Australia. NHLBI, Framingham Heart Study, Framingham, MA USA. Harvard Univ, Sch Med, Massachusetts Gen Hosp, Boston, MA USA. Boston Univ, Dept Math, Boston, MA 02215 USA. RP Lindpaintner, K (reprint author), Harvard Univ, Sch Med, Brigham & Womens Hosp, Div Cardiovasc, 75 Francis St,Thorn 1203, Boston, MA 02115 USA. FU NHLBI NIH HHS [K04-HL-03138-01, R01-HC38038] NR 44 TC 45 Z9 52 U1 1 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1079-5642 J9 ARTERIOSCL THROM VAS JI Arterioscler. Thromb. Vasc. Biol. PD FEB PY 2000 VL 20 IS 2 BP 593 EP 600 PG 8 WC Hematology; Peripheral Vascular Disease SC Hematology; Cardiovascular System & Cardiology GA 281UM UT WOS:000085179200042 PM 10669660 ER PT J AU Couture, P Otvos, JD Cupples, LA Wilson, PWF Schaefer, EJ Ordovas, JM AF Couture, P Otvos, JD Cupples, LA Wilson, PWF Schaefer, EJ Ordovas, JM TI Absence of association between genetic variation in the promoter of the microsomal triglyceride transfer protein gene and plasma lipoproteins in the Framingham Offspring Study SO ATHEROSCLEROSIS LA English DT Article DE microsomal triglyceride transfer protein; LDL-cholesterol; lipoprotein subfractions ID B-CONTAINING LIPOPROTEINS; CORONARY-ARTERY DISEASE; APOLIPOPROTEIN-B; HEART-DISEASE; HEPG2 CELLS; MTP GENE; ABETALIPOPROTEINEMIA; CHOLESTEROL; EXPRESSION; SECRETION AB Microsomal triglyceride transfer protein (MTP) is a lipid transfer protein that is required for the assembly and secretion of very low density lipoproteins (VLDL) by the liver and chylomicrons by the intestine. The common G-493T polymorphism of the MTP promoter has been shown to be associated with decreased plasma LDL-cholesterol and ApoB content of VLDL. The purpose of the present study was, therefore, to investigate the association of this mutation with variations in lipid and apoprotein levels, lipoprotein subclass profiles and coronary heart diasease (CHD) risk in a population-based sample of 1226 male and 1284 female Framingham Offspring participants. In men and women, no significant association was found between the G-493T MTP polymorphism and variations of plasma levels of total cholesterol, LDL-cholesterol, apoprotein B, HDL-cholesterol, apoprotein AI and triglycerides. In order to further investigate potential relationships with variations of lipoprotein phenotypes, lipoprotein subclass profiles were measured using automated nuclear magnetic resonance (NMR) spectroscopy. Each NMR profile yielded information on lipid mass of VLDL, LDL, and HDL subclasses. In both genders, there was no significant association between the G-493T polymorphism and variability of lipoprotein subclass distributions or lipoprotein particle size. Furthermore, no significant association was found between the polymorphism of the MTP promoter and prevalence or the age of onset of CHD. Thus, our results suggest that the G-493T mutation in the MTP promoter is unlikely to have significant implications for cardiovascular disease in men and women. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved. C1 Tufts Univ, Jean Mayer USDA Human Nutr Res Ctr Aging, Lipid Metab Lab, Boston, MA 02111 USA. N Carolina State Univ, Dept Biochem, Raleigh, NC 27695 USA. Boston Univ, Sch Publ Hlth, NHLBI, Framingham, MA 01701 USA. NHLBI, Framington Heart Study, Framingham, MA 01701 USA. RP Ordovas, JM (reprint author), Tufts Univ, Jean Mayer USDA Human Nutr Res Ctr Aging, Lipid Metab Lab, 711 Washington St, Boston, MA 02111 USA. OI Ordovas, Jose/0000-0002-7581-5680 FU NHLBI NIH HHS [HL35243, HL54776]; PHS HHS [N01-38038] NR 37 TC 41 Z9 44 U1 0 U2 0 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0021-9150 J9 ATHEROSCLEROSIS JI Atherosclerosis PD FEB PY 2000 VL 148 IS 2 BP 337 EP 343 DI 10.1016/S0021-9150(99)00281-6 PG 7 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 289DR UT WOS:000085605700015 PM 10657570 ER PT J AU Zajac-Kaye, M Ben-Baruch, N Kastanos, E Kaye, FJ Allegra, C AF Zajac-Kaye, M Ben-Baruch, N Kastanos, E Kaye, FJ Allegra, C TI Induction of Myc-intron-binding polypeptides MIBP1 and RFX1 during retinoic acid-mediated differentiation of haemopoietic cells SO BIOCHEMICAL JOURNAL LA English DT Article DE c-myc oncogene; granulocytic differentiation; HL60 cell line; RFX gene; transcription ID HEPATITIS-B VIRUS; C-MYC; COOPERATIVE BINDING; REGULATORY ELEMENT; NUCLEAR-PROTEIN; ENHANCER REGION; DOWN-REGULATION; HL60 CELLS; RXR-ALPHA; IN-VIVO AB Retinoic acid-mediated differentiation of HL60 cells is associated with an alteration of chromatin structure that maps to protein-binding sequences within intron I of the c-myc gene and with down-regulation of c-myc expression. By using HeLa cell extracts, we previously identified two polypeptides, designated MIBP1 (for Myc-intron-binding peptide) and RFX1, that interact in vivo and bind to the intron I element; we showed that tandem repeats of an MIBP1/RFX1-binding site can exhibit silencer activity on a heterologous promoter. Here we demonstrate that p160 MIBP1 and p130 RFX1 are absent from undifferentiated HL60 cells. In addition, we show that treatment with retinoic acid induces both MIBP1 and RFX1 protein, as well as their DNA-binding activity, upon granulocytic differentiation of HL60 cells, with a gel mobility pattern identical to that of HeLa cells. In the absence of p160 MIBP1 and p130 RFX1, we observed that the altered gel mobility-shift pattern detected in undifferentiated HL60 cells reflects the binding of two novel polypeptides, p30 and p97, that can be cross-linked to the same recognition intron sequence. We also show that the time course of MIBP1 and RFX1 induction is inversely correlated with the down-regulation of c-myc levels during the treatment of HL60 cells with retinoic acid. C1 NCI, Med Branch, Dept Dev Therapeut, Div Clin Sci, Bethesda, MD 20889 USA. NCI, Div Clin Sci, Med Branch, Dept Genet, Bethesda, MD 20889 USA. RP Zajac-Kaye, M (reprint author), NCI, Med Branch, Dept Dev Therapeut, Div Clin Sci, Bethesda, MD 20889 USA. EM Kayem@exchange.nih.gov RI kaye, frederic/E-2437-2011 NR 45 TC 17 Z9 19 U1 0 U2 1 PU PORTLAND PRESS PI LONDON PA 59 PORTLAND PLACE, LONDON W1N 3AJ, ENGLAND SN 0264-6021 J9 BIOCHEM J JI Biochem. J. PD FEB 1 PY 2000 VL 345 BP 535 EP 541 DI 10.1042/0264-6021:3450535 PN 3 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 285FN UT WOS:000085379200018 PM 10642512 ER PT J AU Hill, RM Parmar, PK Coates, LC Mezey, E Pearson, JF Birch, NP AF Hill, RM Parmar, PK Coates, LC Mezey, E Pearson, JF Birch, NP TI Neuroserpin is expressed in the pituitary and adrenal glands and induces the extension of neurite-like processes in AtT-20 cells SO BIOCHEMICAL JOURNAL LA English DT Article DE endocrine; serpin; tissue plasminogen activator ID TISSUE-PLASMINOGEN ACTIVATOR; NERVE GROWTH-FACTOR; EXTRACELLULAR-MATRIX; CLINICAL ASPECTS; MOLECULAR-FORMS; FIBER GROWTH; INHIBITOR; MODULATION; VESICLES; NEURONS AB Two cDNAs encoding the serine protease inhibitor (serpin) neuroserpin were cloned from a rat pituitary cDNA library (rNS-1, 2922 bp; rNS-2, 1599 bp). In situ hybridization histochemistry showed neuroserpin transcripts in the intermediate, anterior and posterior lobes of the pituitary gland and medullary cells in the adrenal gland. Expression of rNS-1 mRNA was restricted to selected cells in the pituitary gland. Analysis of purified secretory-granule fractions from pituitary and adrenal tissues indicated that neuroserpin was found in dense-cored secretory granules. This result suggested that endocrine neuroserpin may regulate intragranular proteases or inhibit enzymes following regulated secretion. To investigate the function of neuroserpin in endocrine tissues we established stable anterior pituitary AtT-20 cell lines expressing neuroserpin. Cells with increased levels of neuroserpin responded by extending neurite-like processes. Extracellular proteolysis by serine protease plasminogen activators has been suggested to regulate neurite outgrowth. As neuroserpin inhibits tissue plasminogen activator (tPA) in vitro, we measured plasminogen-activator levels. Zymographic analysis indicated that AtT-20 cells synthesized and secreted a plasminogen activator identical in size to tPA. A higher-molecular-mass tPA-neuroserpin complex was also observed in AtT-20-cell conditioned culture medium. tPA levels were similar in parent AtT-20 cells and a stable cell line with increased levels of neuroserpin. There was no accumulation of a tPA-neuroserpin complex. Together these results identify endocrine cells as an important source of neuroserpin. Moreover they suggest that neuroserpin is released from dense-cored secretory granules to regulate cell-extracellular matrix interactions through a mechanism that may not directly involve tPA. C1 Univ Auckland, Sch Biol Sci, Mol Neuroendocrinol Lab, Auckland 1, New Zealand. NINDS, Basic Neurosci Program, NIH, Bethesda, MD 20892 USA. Univ Auckland, Sch Math & Informat Sci, Dept Stat, Auckland 1, New Zealand. RP Birch, NP (reprint author), Univ Auckland, Sch Biol Sci, Mol Neuroendocrinol Lab, Private Bag 92019, Auckland 1, New Zealand. RI Birch, Nigel/H-2498-2011; Pearson, John/B-4376-2012; pearson, john/D-7428-2012; OI Birch, Nigel/0000-0002-8417-3587; Pearson, John/0000-0001-5607-4517 NR 43 TC 35 Z9 37 U1 1 U2 1 PU PORTLAND PRESS LTD PI LONDON PA THIRD FLOOR, EAGLE HOUSE, 16 PROCTER STREET, LONDON WC1V 6 NX, ENGLAND SN 0264-6021 J9 BIOCHEM J JI Biochem. J. PD FEB 1 PY 2000 VL 345 BP 595 EP 601 DI 10.1042/0264-6021:3450595 PN 3 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 285FN UT WOS:000085379200024 PM 10642518 ER PT J AU Barry, CE Slayden, RA Sampson, AE Lee, RE AF Barry, CE Slayden, RA Sampson, AE Lee, RE TI Use of genomics and combinatorial chemistry in the development of new antimycobacterial drugs SO BIOCHEMICAL PHARMACOLOGY LA English DT Article DE tuberculosis; isoniazid; mycobacteria; drug design; Mycobacterium tuberculosis; genomics; microarray ID RESISTANT MYCOBACTERIUM-TUBERCULOSIS; ISONIAZID-RESISTANT; MURINE TUBERCULOSIS; GENE-EXPRESSION; MUTATIONS; PYRAZINAMIDE; TARGET; KATG; METRONIDAZOLE; ETHIONAMIDE AB With the completion of the genome of Mycobacterium tuberculosis comes the promise of a new generation of potent drugs to combat the emerging epidemic of multiply drug-resistant isolates. Translating this genomic information into realistic assays, valid targets, and preclinical drug candidates represents the next great hope in tuberculosis control. We propose a paradigm for exploiting the genome to inform the development of novel antituberculars, utilizing the techniques of differential gene expression as monitored by DNA microarrays coupled with the emerging discipline of combinatorial chemistry. A comparison of currently used antituberculars with the properties of other pharmaceuticals suggests that such compounds will have a defined range of physiochemical properties. In general, we can expect the next generation of antituberculars to be small, relatively hydrophilic molecules that bind tightly to specific cellular targets. Many current antimycobacterials require some form of cellular activation (e.g. the activation of isoniazid by a catalase-peroxidase). Activation corresponds to the oxidative, reductive, or hydrolytic unmasking of reactive groups, which occurs with many current antimycobacterial prodrugs. Understanding the mechanisms involved in activation of current antimycobacterial therapeutics also may facilitate the development of alternative activation strategies or of analogs that require no such processes. (C) 1999 Elsevier Science Inc. C1 NIAID, TB Res Sec, Host Def Lab, NIH, Rockville, MD 20852 USA. RP Barry, CE (reprint author), NIAID, TB Res Sec, Host Def Lab, NIH, 12441 Parklawn Dr, Rockville, MD 20852 USA. EM clifton_barry@nih.gov RI Barry, III, Clifton/H-3839-2012; Lee, Richard/J-4997-2013; Slayden, Richard/O-8626-2016 OI Lee, Richard/0000-0002-2397-0443; Slayden, Richard/0000-0001-6857-7277 FU Intramural NIH HHS [Z01 AI000693-15] NR 84 TC 97 Z9 99 U1 0 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0006-2952 J9 BIOCHEM PHARMACOL JI Biochem. Pharmacol. PD FEB 1 PY 2000 VL 59 IS 3 BP 221 EP 231 DI 10.1016/S0006-2952(99)00253-1 PG 11 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 265VU UT WOS:000084267300002 PM 10609550 ER PT J AU Kodama, EN McCaffrey, RP Yusa, K Mitsuya, H AF Kodama, EN McCaffrey, RP Yusa, K Mitsuya, H TI Antileukemic activity and mechanism of action of cordycepin against terminal deoxynucleotidyl transferase-positive (TdT(+)) leukemic cells SO BIOCHEMICAL PHARMACOLOGY LA English DT Article DE cordycepin; adenosine deaminase; antileukemic agent; terminal deoxynucleotidyl transferase ID DEAMINASE INHIBITOR 2'-DEOXYCOFORMYCIN; ADENOSINE-DEAMINASE; PROGNOSTIC-SIGNIFICANCE; B-CELLS; 2',3'-DIDEOXYADENOSINE; THYMOCYTES; EXPRESSION; INSERTION; MESSENGER AB The nucleoside analogue cordycepin (3'-deoxyadenosine, 3'-dA) is substantially more cytotoxic to terminal deoxynucleotidyl transferase positive (TdT(+)) leukemic cells than to TdT(-) leukemic cells in vitro in the presence of an adenosine deaminase inhibitor, deoxycoformycin (dCF), and has been considered as a therapeutic agent for TdT(+) leukemia. The intracellular metabolism of 3'-dA was examined with HPLC, and the mechanism of its anti-TdT(+) leukemic activity was analyzed. In the presence of dCF (2.5 mu M), TdT(+) leukemic cells (N = 5) were sensitive to the cytotoxic effect of 3' dA, whereas TdT(-) (N = 6) cells were not. A high level of 3'-dA-5'-triphosphate (3'-dATP) formation was detected in TdT(+) NALM-6 cells (67 pmol/10(6) cells) and TdT(-) K562 cells (49 pmol/10(6) cells) when cultured with 1 mu M [3'-H-3]-labeled 3'-dA. A substantial level of 3'-dATP was detected in TdT(-) HUT-102 cells (27 pmol/10(6) cells), whereas the level of 3'-dATP in TdT(+) MOLT-4 cells was low (0.3 pmol/10(6) cells). The mean Ic(50) values of 3'-dA against phytohemagglutinin (PHA)-activated and resting peripheral blood mononuclear cells (PBM) (N = 5) were 8 and 32 mu M, respectively. There was a modest level of 3'-dATP (7 pmol/10(6) cells) in PHA-PBM, whereas a lower level of 3'-dATP was detected in resting PBM (2.5 pmol/10(6) cells). These data suggest that the presence of 3'-dATP is not sufficient for the antileukemic effect of 3'-dA, but that TdT positivity is essential, and that PBM are significantly less sensitive to the cytotoxicity of 3' dA in vitro. Further development of 3'-dA as a potential antileukemic agent to treat patients with TdT(+) leukemia is warranted. (C) 1999 Elsevier Science Inc. C1 NCI, Expt Retrovirol Sect, Med Branch, NIH, Bethesda, MD 20892 USA. Boston Univ, Med Ctr, Martin H Semler Lab, Sect Med Oncol,Evans Mem Dept Clin Res, Boston, MA 02118 USA. Kumamoto Univ, Sch Med, Dept Internal Med 2, Kumamoto 860, Japan. RP Mitsuya, H (reprint author), NCI, Expt Retrovirol Sect, Med Branch, NIH, 9000 Rockville Pike, Bethesda, MD 20892 USA. RI Kodama, Eiichi /C-4032-2009 OI Kodama, Eiichi /0000-0002-6622-2752 NR 40 TC 78 Z9 105 U1 0 U2 5 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0006-2952 J9 BIOCHEM PHARMACOL JI Biochem. Pharmacol. PD FEB 1 PY 2000 VL 59 IS 3 BP 273 EP 281 DI 10.1016/S0006-2952(99)00325-1 PG 9 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 265VU UT WOS:000084267300008 PM 10609556 ER PT J AU Sharp, PM Bailes, E Gao, F Beer, BE Hirsch, VM Hahn, BH AF Sharp, PM Bailes, E Gao, F Beer, BE Hirsch, VM Hahn, BH TI Origins and evolution of AIDS viruses: estimating the time-scale SO BIOCHEMICAL SOCIETY TRANSACTIONS LA English DT Article; Proceedings Paper CT Colloquium on Evolution of Sequences, Structures and Genomes CY SEP 07-09, 1999 CL UNIV COLL CORK, CORK, IRELAND HO UNIV COLL CORK DE gamma distribution; HIV-I; host-dependent evolution; molecular clock primate lentiviruses ID SIMIAN IMMUNODEFICIENCY VIRUS; CROSS-SPECIES TRANSMISSION; AFRICAN-GREEN MONKEYS; MOLECULAR CLOCK; IN-VIVO; PHYLOGENETIC ANALYSIS; SEQUENCE; HIV-1; INFECTION; TYPE-1 AB The primate lentiviruses comprise SIV strains from various host species, as well as two viruses, HIV-1 and HIV-2, that cause AIDS in humans. The origins of HIV-1 and HIV-2 have been traced to cross-species transmissions from chimpanzees and sooty mangabey monkeys respectively. Two approaches have been taken to estimate the timescale of the evolution of these viruses. Certain groups of SIV strains appear to have evolved in a host-dependent manner, implying a time-scale of many thousands or even millions of years. In stark contrast, molecular clock calculations have previously been used to estimate a time-scale of only tens or hundreds of years. Those calculations largely ignored heterogeneity of evolutionary rates across different sites within sequences. In fact, the distribution of rates at different sites seems extremely skewed in HIV-1, and so the time-depth of the primate lentivirus evolutionary tree may have been underestimated by at least a factor of ten. However, these date estimates still seem to be far too recent to be consistent with host-dependent evolution. C1 Univ Nottingham, Queens Med Ctr, Genet Inst, Nottingham NG7 2UH, England. Univ Alabama, Dept Med, Birmingham, AL 35294 USA. Univ Alabama, Dept Microbiol, Birmingham, AL 35294 USA. NIAID, Mol Microbiol Lab, NIH, Rockville, MD 20852 USA. RP Sharp, PM (reprint author), Univ Nottingham, Queens Med Ctr, Genet Inst, Nottingham NG7 2UH, England. RI Sharp, Paul/F-5783-2010 OI Sharp, Paul/0000-0001-9771-543X NR 46 TC 99 Z9 100 U1 4 U2 21 PU PORTLAND PRESS PI LONDON PA 59 PORTLAND PLACE, LONDON W1N 3AJ, ENGLAND SN 0300-5127 J9 BIOCHEM SOC T JI Biochem. Soc. Trans. PD FEB PY 2000 VL 28 BP 275 EP 282 PN 2 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 284CL UT WOS:000085313100054 PM 10816142 ER PT J AU Haltiwanger, BM Matsumoto, Y Nicolas, E Dianov, GL Bohr, VA Taraschi, TF AF Haltiwanger, BM Matsumoto, Y Nicolas, E Dianov, GL Bohr, VA Taraschi, TF TI DNA base excision repair in human malaria parasites is predominantly by a long-patch pathway SO BIOCHEMISTRY LA English DT Article ID CELL-FREE-EXTRACTS; PLASMODIUM-FALCIPARUM; POLYMERASE-BETA; DEOXYRIBONUCLEIC ACID; BOVINE TESTIS; SITE REPAIR; LIGASE-I; RECONSTITUTION; REQUIREMENT; PROTEIN AB Mammalian cells repair apurinic/apyrimidinic (AP) sites in DNA by two distinct pathways: a polymerase beta (pol beta)-dependent, short- (one nucleotide) patch base excision repair (BER) pathway, which is the major route, and a PCNA-dependent, long- (several nucleotide) patch BER pathway. The ability of a cell-free lysate prepared from asexual Plasmodium falciparum malaria parasites to remove uracil and repair AP sites in a variety of DNA substrates was investigated. We found that the lysate contained uracil DNA glycosylase, AP endonuclease, DNA polymerase, flap endonuclease, and DNA ligase activities. This cell-free lysate effectively repaired a regular or synthetic AP site on a covalently closed circular (ccc) duplex plasmid molecule or a long (382 bp), Linear duplex DNA fragment, or a regular or reduced AP site in short (28 bp), duplex oligonucleotides. Repair of the AP sites in the various DNA substrates involved a long-patch BER pathway. This biology is different from mammalian cells, yeast, Xenopus, and Escherichia coli, which predominantly repair AP sites by a one-nucleotide patch BER pathway. The apparent absence of a short-patch BER pathway in P. falciparum may provide opportunities to develop antimalarial chemotherapeutic strategies for selectively damaging the parasites in vivo and will allow the characterization of the long-patch BER pathway without having to knock-out or inactivate a short-patch BER pathway, which is necessary in mammalian cells. C1 Thomas Jefferson Univ, Dept Microbiol & Mol Virol, Philadelphia, PA 19107 USA. Fox Chase Canc Ctr, Dept Radiat Oncol, Philadelphia, PA 19111 USA. Thomas Jefferson Univ, Dept Pathol Anat & Cell Biol, Philadelphia, PA 19107 USA. NIA, Genet Mol Lab, NIH, Baltimore, MD 21224 USA. RP Taraschi, TF (reprint author), Thomas Jefferson Univ, Dept Microbiol & Mol Virol, 1020 Locust St, Philadelphia, PA 19107 USA. FU NIAID NIH HHS [AI41761] NR 40 TC 30 Z9 31 U1 0 U2 0 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD FEB 1 PY 2000 VL 39 IS 4 BP 763 EP 772 DI 10.1021/bi9923151 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 278WZ UT WOS:000085013100017 PM 10651642 ER PT J AU Narayan, S Wilson, SH AF Narayan, S Wilson, SH TI Kinetic analysis of Sp1-mediated transcriptional activation of a TATA-Containing promoter SO BIOCHEMISTRY LA English DT Article ID RNA-POLYMERASE-II; OPEN COMPLEX-FORMATION; BOX-BINDING-PROTEINS; PREINITIATION COMPLEX; FUNCTIONAL-ANALYSIS; TFIID COMPLEX; PRM PROMOTER; SP1; INITIATION; MECHANISM AB Using a HeLa cell nuclear extract (NE)-based in vitro runoff transcription system, we have examined the effect of Spl on the activation of a TATA-containing chimeric DNA polymerase beta (pAS8) promoter. The results demonstrated that the TATA element-dependent basal activity of the pAS8 promoter was stimulated 4-fold by supplementation of a Sp1-depleted HeLa cell nuclear extract (NEd) with purified human Spl, indicating that pAS8 promoter activity is dependent upon Spl. A detailed kinetic analysis based on a three-step kinetic model of transcription initiation showed that Sp1 stimulates the activity of the pAS8 promoter by increasing the amount of closed preinitiation complex (RPc) assembly as well as by enhancing the rate of promoter clearance (k(3)). Then was no significant effect of Spl on the apparent rate of open complex (RPo) formation (k(2)) of the pAS8 promoter. These studies define more precisely the kinetic mechanisms by which Sp1 may regulate the rate of transcript formation of a TATA-containing promoter. C1 NIEHS, Struct Biol Lab, NIH, Res Triangle Pk, NC 27709 USA. Univ Texas, Med Branch, Sealy Ctr Mol Sci, Galveston, TX 77555 USA. Univ Texas, Med Branch, Sealy Ctr Oncol & Hematol, Galveston, TX 77555 USA. RP Wilson, SH (reprint author), NIEHS, Struct Biol Lab, NIH, Res Triangle Pk, NC 27709 USA. FU NCI NIH HHS [CA77721]; NIEHS NIH HHS [ES06492] NR 40 TC 6 Z9 6 U1 0 U2 1 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0006-2960 J9 BIOCHEMISTRY-US JI Biochemistry PD FEB 1 PY 2000 VL 39 IS 4 BP 818 EP 823 DI 10.1021/bi9912701 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 278WZ UT WOS:000085013100023 PM 10651648 ER PT J AU Masuda, S Kamao, M Schroeder, NJ Makin, HLJ Jones, G Kremer, R Rhim, J Okano, T AF Masuda, S Kamao, M Schroeder, NJ Makin, HLJ Jones, G Kremer, R Rhim, J Okano, T TI Characterization of 3-epi-1 alpha,25-dihydroxyvitamin D-3 involved in 1 alpha,25-dihydroxyvitamin D-3 metabolic pathway in cultured cell lines SO BIOLOGICAL & PHARMACEUTICAL BULLETIN LA English DT Article DE 3-epi-1 alpha,25-dihydroxyvitamin D-3; 1 alpha,25-dihydroxyvitamin D-3; C24-oxidation; bone; C3-epimerization ID A-RING DIASTEREOMERS; IN-VITRO METABOLISM; D ENDOCRINE SYSTEM; VITAMIN-D ANALOG; 1,25-DIHYDROXYVITAMIN-D3; ENZYME AB Using sis different cultured cell models representing osteoblast, intestine, kidney and keratinocyte, we have demonstrated that 1 alpha,25-dihydroxyvitamin D-3 (1 alpha,25(OH)(2)D-3) is metabolized into 3-epi-1 alpha,25(OH)(2)D-3 in vitamin D-target cells. Although differences existed in the amount of 3-epi-1 alpha,25(OH)(2)D-3 formed with different cell types, it was apparent that 1 alpha,25(OH)(2)D-3 was subjected to metabolism both through the C24-oxidation and 3-epimerization pathways. Time course and dose response studies showed that the production of 3-epi-1 alpha,25(OH)(2)D-3 was enzymatic. It is interesting to note that this epimerization proceeded from 3 beta towards 3 or unidirectionally, and this conversion was not inhibited by ketoconazole. These data suggest that cytochrome P450 related enzymes including the 24-hydroxylase would not affect this reaction. The biological activity of J-epi-1 alpha,25(OH)(2)D-3 was found to be lower than the native 1 alpha,25(OH)(2)D-3 in suppressing of proliferation of HL-60 cells, while the affinity of 3-epi-1 alpha,25(OH)(2)D-3 for vitamin D-binding protein was 2.5-fold higher than that of 1 alpha,25(OH)(2)D-3. The results indicate that 3-epimerization may change the pharmacokinetics and catabolism of 1 alpha,25(OH)(2)D-3 in vitamin D-target cells. C1 Kobe Pharmaceut Univ, Dept Hyg Sci, Higashinada Ku, Kobe, Hyogo 6588558, Japan. St Bartholomews & Royal London Sch Med & Dent, Dept Clin Biochem, London E1 2AD, England. Queens Univ, Dept Biochem, Kingston, ON K7L 3N6, Canada. Royal Victoria Hosp, Calcium Res Lab, Montreal, PQ H3A 1A1, Canada. NCI, Lab Biochem Physiol, Frederick, MD 21702 USA. RP Masuda, S (reprint author), Coll Nursing Art & Sci, Dept Biochem & Physiol, 13-71 Kitaoji Cho, Akashi, Hyogo 6588588, Japan. NR 22 TC 24 Z9 25 U1 0 U2 0 PU PHARMACEUTICAL SOC JAPAN PI TOKYO PA 2-12-15 SHIBUYA, SHIBUYA-KU, TOKYO, 150-0002, JAPAN SN 0918-6158 J9 BIOL PHARM BULL JI Biol. Pharm. Bull. PD FEB PY 2000 VL 23 IS 2 BP 133 EP 139 PG 7 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 282TA UT WOS:000085233700001 PM 10706373 ER PT J AU Jefferson, WN Couse, JF Banks, EP Korach, KS Newbold, RR AF Jefferson, WN Couse, JF Banks, EP Korach, KS Newbold, RR TI Expression of estrogen receptor beta is developmentally regulated in reproductive tissues of male and female mice SO BIOLOGY OF REPRODUCTION LA English DT Article ID MESSENGER-RIBONUCLEIC-ACID; WILD-TYPE; ER-BETA; CHROMOSOMAL LOCALIZATION; BINDING PROTEIN; RNA-BINDING; MOUSE; DIETHYLSTILBESTROL; ALPHA; TRACT AB By the use of ribonuclease protection assay (RPA) combined with immunohistochemical techniques, the expression of estrogen receptor (ER) or and ERP was mapped in the developing gonads and reproductive tracts of male and female mice from fetal day 14 to postnatal day 26 (PND 26), This study was designed to determine the pattern of expression of both ER subtypes in specific tissue compartments during development. In ovaries, ER alpha mRNA was detected at all ages examined; ER beta mRNA was seen as early as PND 1, and its expression increased with age. Immunolocalization showed ER beta in differentiating granulosa cells of the ovary, whereas ER alpha was predominantly seen in interstitial cells. The remainder of the female reproductive tract showed ER alpha mRNA at all ages examined with little or no significant levels of ER beta, except on END 1 when a low level of message appeared. In males, ER alpha and ERP mRNA were detected in the fetal testis; however, ER beta gradually increased until PND 5 and subsequently diminished to undetectable levels by PND 26. Immunolocalization showed ER alpha in the interstitial compartment of the testis, whereas ER beta was seen predominantly in developing spermatogonia, The remainder of the male reproductive tract showed varying amounts of both receptors by RPA and immunostaining throughout development. These studies provide information useful in studying the role of both ER subtypes in normal differentiation, and they provide indications of differential tissue expression during development. C1 NIEHS, Dev Endocrinol Sect, Res Triangle Pk, NC 27709 USA. NIEHS, Reprot Toxicol Grp, Toxicol Lab, Environm Toxicol Program, Res Triangle Pk, NC 27709 USA. NIEHS, Receptor Biol Sect, Lab Reprod & Dev Toxicol, Environm Dis & Med Program, Res Triangle Pk, NC 27709 USA. RP Newbold, RR (reprint author), NIEHS, Dev Endocrinol Sect, Mail Drop E4-02,111 Alexander Dr,South Campus,POB, Res Triangle Pk, NC 27709 USA. OI Korach, Kenneth/0000-0002-7765-418X NR 36 TC 145 Z9 157 U1 0 U2 1 PU SOC STUDY REPRODUCTION PI MADISON PA 1603 MONROE ST, MADISON, WI 53711-2021 USA SN 0006-3363 J9 BIOL REPROD JI Biol. Reprod. PD FEB PY 2000 VL 62 IS 2 BP 310 EP 317 DI 10.1095/biolreprod62.2.310 PG 8 WC Reproductive Biology SC Reproductive Biology GA 278JV UT WOS:000084987400012 PM 10642567 ER PT J AU Haverkos, H Rohrer, M Pickworth, W AF Haverkos, H Rohrer, M Pickworth, W TI The cause of invasive cervical cancer could be multifactorial SO BIOMEDICINE & PHARMACOTHERAPY LA English DT Article DE cervical cancer; human papillomavirus; tars ID HUMAN-PAPILLOMAVIRUS INFECTION; HUMAN-IMMUNODEFICIENCY-VIRUS; ORAL-CONTRACEPTIVE USE; RISK-FACTORS; CIGARETTE-SMOKING; TYPE-2; WOMEN; DNA; DYSPLASIA; NEOPLASIA AB Cancer of the cervix is the third most common cancer among women worldwide. with incidence rates ranging from 3.8 per 100.000 women per year in Israel to 48.2 per 100.000 per year in Colombia. Epidemiologic and clinical data suggest that human papillomaviruses. especially HPV-16 and HPV-18, play the major role in the etiology of cervical cancer. However, many investigators acknowledge that HPV is neither necessary nor sufficient in the etiology or cervical cancer and that a mulifactorial etiology is likely. HPV cannot be found in every patient with the disease and other factor. such us herpes simplex virus type 2 infection. cigarette smoking, vaginal douching. nutrition, and use of oral contraceptives, have been associated with cervical cancer. In two different animal models. tumors can be produced following exposure to DNA viruses and tars. Using those animal models as prototypes. we propose that the etiology of cervical cancer in humans could he an interaction between DNA viruses. specifically papillomavirus and/or HSV-2 infection, and tar exposure through cigarette smoking and/or tar-based vaginal douching. (C) 2000 Editions scientifiques et medicales Elsevier SAS. C1 Walter Reed Army Med Ctr, Dept Med, Div Infect Dis, Washington, DC 20307 USA. NIDA, Addict Res Ctr, NIH, Baltimore, MD 21224 USA. RP Haverkos, H (reprint author), HFD-530,5600 Fishers Lane, Rockville, MD 20857 USA. NR 47 TC 34 Z9 38 U1 0 U2 3 PU EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER PI PARIS CEDEX 15 PA 23 RUE LINOIS, 75724 PARIS CEDEX 15, FRANCE SN 0753-3322 J9 BIOMED PHARMACOTHER JI Biomed. Pharmacother. PD FEB PY 2000 VL 54 IS 1 BP 54 EP 59 DI 10.1016/S0753-3322(00)88642-4 PG 6 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 291NA UT WOS:000085741900010 PM 10721464 ER PT J AU Bevan, DR Li, LP Pedersen, LG Darden, TA AF Bevan, DR Li, LP Pedersen, LG Darden, TA TI Molecular dynamics simulations of the d(CCAACGTTGG)(2) decamer: Influence of the crystal environment SO BIOPHYSICAL JOURNAL LA English DT Article ID PARTICLE-MESH-EWALD; B-DNA; NUCLEIC-ACIDS; A-DNA; MINOR-GROOVE; FORCE-FIELD; HYDRATION; SEQUENCE; WATER; D(CGCGAATTCGCG) AB Molecular dynamics (MD) simulations of the DNA duplex d(CCAACGTTGG)(2) were used to study the relationship between DNA sequence and structure. Two crystal simulations were carried out; one consisted of one unit cell containing two duplexes, and the other of two unit cells containing four duplexes. Two solution simulations were also carried out, one starting from canonical B-DNA and the other starting from the crystal structure, For many helicoidal parameters, the results from the crystal and solution simulations were essentially identical. However, for other parameters, in particular, alpha, gamma, delta, (epsilon - zeta), phase, and helical twist, differences between crystal and solution simulations were apparent. Notably, during crystal simulations, values of helical twist remained comparable to those in the crystal structure, to include the sequence-dependent differences among base steps, in which values ranged from 20 degrees to 50 degrees per base step, However, in the solution simulations, not only did the average values of helical twist decrease to similar to 30 degrees per base step, but every base step was similar to 30 degrees, suggesting that the sequence-dependent information may be lost. This study reveals that MD simulations of the crystal environment complement solution simulations in validating the applicability of MD to the analysis of DNA structure. C1 Virginia Polytech Inst & State Univ, Dept Biochem, Blacksburg, VA 24061 USA. NIEHS, Struct Biol Lab, Res Triangle Pk, NC 27709 USA. RP Bevan, DR (reprint author), Virginia Tech, Dept Biochem, 201 Fralin Biotechnol Ctr, Blacksburg, VA 24061 USA. EM drbevan@vt.edu RI Pedersen, Lee/E-3405-2013 OI Pedersen, Lee/0000-0003-1262-9861 NR 47 TC 46 Z9 47 U1 0 U2 9 PU CELL PRESS PI CAMBRIDGE PA 600 TECHNOLOGY SQUARE, 5TH FLOOR, CAMBRIDGE, MA 02139 USA SN 0006-3495 J9 BIOPHYS J JI Biophys. J. PD FEB PY 2000 VL 78 IS 2 BP 668 EP 682 PG 15 WC Biophysics SC Biophysics GA 282YL UT WOS:000085249300011 PM 10653780 ER PT J AU Shi, CS Tuscano, J Kehrl, JH AF Shi, CS Tuscano, J Kehrl, JH TI Adaptor proteins CRK and CRKL associate with the serine/threonine protein kinase GCKR promoting GCKR and SAPK activation SO BLOOD LA English DT Article ID GERMINAL CENTER KINASE; N-TERMINAL KINASE; SH3 DOMAIN; V-CRK; PATHWAY; HPK1; CASCADE; FAMILY; RAS; C3G AB STE20-related kinases play significant regulatory roles in a range of cellular responses to environmental stimuli. GCKR (also referred to as KHS1) is a serine/threonine protein kinase that has an STE20-like protein kinase domain and that stimulates the stress-activated protein kinase (SAPK, also referred to as Jun kinase or JNK) pathway. GCKR has a large C-terminal regulatory domain that provides sites for interactions with other proteins. Adaptor proteins mediate the Interactions between signaling molecules. In this study we showed that the adaptor proteins Crk and CrkL associated with GCKR. When Crk-I, Crk-II, or CrkL was transiently expressed In HEK 293T cells along with GCKR, each coimmunoprecipitated with GCKR. Furthermore, in the Bcr-Abl transformed cell line, K562 endogenous GCKR and CrkL coimmunoprecipitated, indicating a constitutive association, Detection of the CrkL-GCKR Interaction required the SH3 domains of CrkL and 2 regions in GCKR - 1 between amino acids 387 and 395 that contains a consensus SH3 binding motif and the other between amino acids 599 and 696, Crk or CrkL overexpression increased GCKR catalytic activity, A dominant negative form of Res abolished Crk- or CrkL-induced GCKR activation, suggesting a dependence on Res activation for their activation of GCKR, Finally, we showed Impairment of the known ability of CrkL to activate the SAPK pathway by a catalytically Inactive form of GCKR or by a GCKR antisense construct. Thus, GCKR associates with other proteins through interactions mediated by SH2/SH3 adaptor proteins, which can lead to GCKR and SAPK activation. (C) 2000 by The American Society of Hematology. C1 NIAID, Immunoregulat Lab, Cell Mol Immunol Sect B, NIH, Bethesda, MD 20892 USA. Univ Calif Davis, Davis Canc Ctr, Dept Med, Sacramento, CA 95817 USA. RP Kehrl, JH (reprint author), NIAID, Immunoregulat Lab, Cell Mol Immunol Sect B, NIH, Bldg 10,Room 11B08,10 Ctr Dr,MSC 1876, Bethesda, MD 20892 USA. OI Kehrl, John/0000-0002-6526-159X NR 34 TC 15 Z9 15 U1 0 U2 0 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1200 19TH ST, NW, STE 300, WASHINGTON, DC 20036-2422 USA SN 0006-4971 J9 BLOOD JI Blood PD FEB 1 PY 2000 VL 95 IS 3 BP 776 EP 782 PG 7 WC Hematology SC Hematology GA 278MN UT WOS:000084993700007 PM 10648385 ER PT J AU Smith, RD Li, J Noguchi, CT Schechter, AN AF Smith, RD Li, J Noguchi, CT Schechter, AN TI Quantitative PCR analysis of HbF inducers in primary human adult erythroid cells SO BLOOD LA English DT Article ID FETAL HEMOGLOBIN PRODUCTION; GLOBIN GENE-EXPRESSION; BETA-THALASSEMIA; GAMMA-GLOBIN; HYDROXYUREA; ANEMIA; 5-AZACYTIDINE; BUTYRATE; INDUCTION; DISEASE AB The development and evaluation of drugs to elevate fetal hemoglobin in the treatment of the genetic diseases of hemoglobin would be facilitated by the availability of reliable cell assays. We have used rear-time, quantitative polymerase chain reaction (PCR) analyses of globin messenger RNA (mRNA) levels in a biphasic, erythropoietin-dependent primary culture system for human adult erythroid cells in order to assay compounds for their ability to modulate levels of adult (beta) and fetal (gamma) globin mRNA, Complementary DNA synthesized from total RNA extracted at timed intervals from aliquots of cells were assayed throughout the period that the culture was studied. gamma-globin mRNA levels were found to be much lower (less than 1%) than beta-globin mRNA levels. At concentrations of agents chosen for minimal effect on cell division, we find that the 3 drugs studied, 5-azacytidine (5 mu mol/L), hydroxyurea (40 mu mol/L), and butyric acid (0.5mmol/L) significantly increase gamma-globin mRNA levels. Interestingly, hydroxyurea also had a small stimulatory effect on beta-globin mRNA levels, while butyric acid caused a twofold inhibition of beta-globin mRNA levels, and 5-azacytidine had little effect on beta-globin mRNA levels. The net result of all 3 drugs was to increase the gamma/(gamma + beta) mRNA ratios by threefold to fivefold. These data suggest that the mechanism is distinct for each drug. The profile of butyric-acid-induced changes on globin gene expression is also quite distinct from changes produced by trichostatin A, a known histone deacetylase inhibitor. Quantitative PCR analyses of human erythroid cells should prove useful for studying the mechanism(s) of action of known inducers of gamma-globin and identifying new drug candidates. (C) 2000 by The American Society of Hematology. C1 NIDDKD, Biol Chem Lab, NIH, Bethesda, MD 20892 USA. RP Schechter, AN (reprint author), NIDDKD, Biol Chem Lab, NIH, Bldg 10,Room 9N307,10 Ctr Dr,MSC 1822, Bethesda, MD 20892 USA. OI Schechter, Alan N/0000-0002-5235-9408 NR 40 TC 52 Z9 53 U1 0 U2 2 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1200 19TH ST, NW, STE 300, WASHINGTON, DC 20036-2422 USA SN 0006-4971 J9 BLOOD JI Blood PD FEB 1 PY 2000 VL 95 IS 3 BP 863 EP 869 PG 7 WC Hematology SC Hematology GA 278MN UT WOS:000084993700019 PM 10648397 ER PT J AU Aokl, Y Tosato, G Nambu, Y Iwamoto, A Yarchoan, R AF Aokl, Y Tosato, G Nambu, Y Iwamoto, A Yarchoan, R TI Detection of vascular endothelial growth factor in AIDS-related primary effusion lymphomas SO BLOOD LA English DT Letter C1 US FDA, Ctr Biol Evaluat & Res, Bethesda, MD 20014 USA. Univ Tokyo, Inst Med Sci, Tokyo, Japan. NCI, HIV & AIDS Malignancy Branch, Div Clin Sci, NIH, Bethesda, MD 20892 USA. RP Aokl, Y (reprint author), US FDA, Ctr Biol Evaluat & Res, Bethesda, MD 20014 USA. NR 5 TC 0 Z9 0 U1 0 U2 0 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1200 19TH ST, NW, STE 300, WASHINGTON, DC 20036-2422 USA SN 0006-4971 J9 BLOOD JI Blood PD FEB 1 PY 2000 VL 95 IS 3 BP 1109 EP 1110 PG 2 WC Hematology SC Hematology GA 278MN UT WOS:000084993700054 ER PT J AU Chanock, SJ Roesler, J Zhan, SX Hopkins, P Lee, P Barrett, DT Christensen, BL Curnutte, JT Gorlach, A AF Chanock, SJ Roesler, J Zhan, SX Hopkins, P Lee, P Barrett, DT Christensen, BL Curnutte, JT Gorlach, A TI Genomic structure of the human p47-phox (NCF1) Gene SO BLOOD CELLS MOLECULES AND DISEASES LA English DT Article DE gene; NADPH-oxidase; chronic granulomatous disease; respiratory burst ID CHRONIC GRANULOMATOUS-DISEASE; RESPIRATORY-BURST OXIDASE; CCAAT DISPLACEMENT PROTEIN; AUTOSOMAL RECESSIVE FORMS; NADPH OXIDASE; CHROMOSOMAL LOCATION; WILLIAMS-SYNDROME; EXPRESSION; ACTIVATION; COMPONENTS AB The cytosolic factor p47-phox, encoded by the NCF1 gene, is an essential component of the phagocyte NADPH-oxidase system. Upon activation of this multicomponent system, p47-phox translocates to the membrane and participates in the electron transfer from NADPH to molecular oxygen. A deficiency or absence of p47-phox is the most common autosomal form of chronic granulomatous disease (CGD). We have cloned and characterized the NCF1 gene from four bacteriophage clones, a P1 clone and genomic DNA from normal individuals. The gene is 15,236 base pairs long and includes 11 exons. It is 98.6% homologous in sequence to at least one pseudogene that maps to the same region of chromosome 7q11.23. Slightly more than half (50.37%) of the wild-type NCF1 gene consists of repetitive elements. In particular, the density of Alu sequences is high (1.4 Alu/kb); there are 21 Alu repeats interspersed through 10 introns. These findings are consistent with the observation that recombination events between the wild-type gene and its highly homologous pseudogenes account for the majority of potentially lethal mutations in p47-phox-deficient chronic granulomatous disease. Analysis of 1.96 kb of sequence 5' of the start of translation revealed a high homology (99.6%) between wild-type and pseudogene clones. Characterization of NCF1 establishes a foundation for detailed molecular analysis of p47-phox-deficient CGD patients as well as for the study of the regulation of the NCF1 gene and pseudogenes, both of which are present as full-length transcripts in normal individuals. C1 NCI, Pediat Oncol Branch, Ctr Adv Technol, Gaithersburg, MD 20877 USA. Genentech Inc, Res Immunol, S San Francisco, CA USA. Scripps Res Inst, Dept Mol & Expt Med, La Jolla, CA USA. RP Chanock, SJ (reprint author), NCI, Pediat Oncol Branch, Ctr Adv Technol, 8717 Grovement Circle, Gaithersburg, MD 20877 USA. RI Gorlach, Agnes/B-3494-2013 FU NIAID NIH HHS [AI 24838] NR 51 TC 13 Z9 15 U1 1 U2 2 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1079-9796 J9 BLOOD CELL MOL DIS JI Blood Cells Mol. Dis. PD FEB PY 2000 VL 26 IS 1 BP 37 EP 46 DI 10.1006/bcmd.2000.0274 PG 10 WC Hematology SC Hematology GA 331GM UT WOS:000088010000005 PM 10772875 ER PT J AU Hentunen, TA Choi, SJ Boyce, BF Dallas, MR Dallas, SL Shen-Ong, GL Roodman, GD AF Hentunen, TA Choi, SJ Boyce, BF Dallas, MR Dallas, SL Shen-Ong, GL Roodman, GD TI A murine model of inflammatory bone disease SO BONE LA English DT Article DE inflammatory bone disease; bone resorption; cytokines; osteomyelitis ID MARROW CULTURES; IN-VIVO; OSTEOCLAST; RESORPTION; HYPERCALCEMIA; MOUSE AB We have recently reported the identification of a new recessive mutation on murine chromosome 18 that results in tail kinks and deformity in the lower extremities of mice. Preliminary examination of the bones of these mice showed that there are abnormalities present that resembled chronic recurrent multifocal osteomyelitis. Accordingly, this new mutation was named "CMO." In this report, we describe the histology of bones in CMO mice, as well as the capacity of the bone marrow cells from these animals to form osteoclasts (OCLs), In addition, we tested conditioned media from nonadherent marrow cells and total. marrow cells from CMO mice for their capacity to induce OCL formation in normal murine marrow cultures. These studies demonstrated that the bone disease in these animals is inflammatory in nature, and a soluble factor(s) that is not IL-1 alpha, IL-6 or TNF-alpha is released by marrow cells from CMO animals and enhances OCL formation in normal murine marrow cultures. (C) 2000 by Elsevier Science Inc. All rights reserved. C1 Audie Murphy VA Hosp, Res Serv 151, San Antonio, TX 78284 USA. Univ Texas, Hlth Sci Ctr, Dept Med Hematol, San Antonio, TX USA. Univ Texas, Hlth Sci Ctr, Dept Pathol, San Antonio, TX USA. Univ Texas, Hlth Sci Ctr, Dept Med Endocrinol, San Antonio, TX USA. NCI, Div Canc Biol, Canc Genet Branch, NIH, Bethesda, MD 20892 USA. RP Roodman, GD (reprint author), Audie Murphy VA Hosp, Res Serv 151, 7400 Merton Minter Blvd, San Antonio, TX 78284 USA. RI Shen-Ong, Grace/C-8327-2014 NR 19 TC 19 Z9 20 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 8756-3282 J9 BONE JI Bone PD FEB PY 2000 VL 26 IS 2 BP 183 EP 188 DI 10.1016/S8756-3282(99)00247-1 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 281LE UT WOS:000085160900013 PM 10678414 ER PT J AU Grafman, J AF Grafman, J TI Picking two scientific roses for the next century SO BRAIN AND COGNITION LA English DT Article C1 NINDS, Cognit Neurosci Sect, NIH, Bethesda, MD 20892 USA. RP Grafman, J (reprint author), NINDS, Cognit Neurosci Sect, NIH, Bldg 10,Room 5C205,MSC 1440, Bethesda, MD 20892 USA. OI Grafman, Jordan H./0000-0001-8645-4457 NR 6 TC 2 Z9 2 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0278-2626 J9 BRAIN COGNITION JI Brain Cogn. PD FEB PY 2000 VL 42 IS 1 BP 10 EP 12 DI 10.1006/brcg.1999.1147 PG 3 WC Neurosciences; Psychology, Experimental SC Neurosciences & Neurology; Psychology GA 299NF UT WOS:000086204200004 PM 10739584 ER PT J AU Schmidt, LA Schulkin, J AF Schmidt, LA Schulkin, J TI Toward a computational affective neuroscience SO BRAIN AND COGNITION LA English DT Article C1 McMaster Univ, Dept Psychol, Hamilton, ON L8S 4K1, Canada. Georgetown Univ, Dept Physiol & Biophys, Washington, DC 20057 USA. NIMH, Clin Neuroendocrinol Branch, Bethesda, MD 20892 USA. RP Schmidt, LA (reprint author), McMaster Univ, Dept Psychol, Hamilton, ON L8S 4K1, Canada. NR 16 TC 5 Z9 5 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0278-2626 J9 BRAIN COGNITION JI Brain Cogn. PD FEB PY 2000 VL 42 IS 1 BP 95 EP 98 DI 10.1006/brcg.1999.1172 PG 4 WC Neurosciences; Psychology, Experimental SC Neurosciences & Neurology; Psychology GA 299NF UT WOS:000086204200029 PM 10739609 ER PT J AU Miyaguchi, K Maeda, Y Collin, C Sihag, RK AF Miyaguchi, K Maeda, Y Collin, C Sihag, RK TI Gene transfer into hippocampal slice cultures with an adenovirus vector driven by cytomegalovirus promoter: Stable co-expression of green fluorescent protein and lacZ genes SO BRAIN RESEARCH BULLETIN LA English DT Article DE gene transfer; hippocampal slice cultures; adenovirus vector; long-term potentiation; astrocytes; pyramidal neurons ID LONG-TERM POTENTIATION; VIRUS; BRAIN; NEURONS; INFECTION; DELIVERY; SYSTEM; MODEL; VIVO; CA1 AB Virus-mediated gene transfer into identified neurons of organotypic hippocampal slice cultures offers a great potential for studying the cellular and molecular mechanisms of synaptic plasticity. We describe here a new adenovirus vector Ad-GFP-lacZ carrying an early cytomegalovirus (CMV) gene promoter that efficiently co-transferred the beta-galactosidase (lacZ) and green fluorescent protein (GFP) genes in rat organotypic hippocampal slice cultures. Monitoring of GFP fluorescence and immuno-histochemical staining for beta-galactosidase showed that the expression of the transferred genes was widespread in the glial cells and neurons of CA1, CA3/4, and dentate gyrus regions. Immunoblot analyses showed that the expression of beta-galactosidase and GFP was maximal about 48 h after infection of hippocampal slices with the adenovirus vector and the expression levels were maintained for several weeks. Also, immunoblot analyses showed no significant differences in the MAP-2 and glial fibrillary acidic protein levels in the adenovirus vector infected and uninfected hippocampal slices. In addition, we found that the infection of hippocampal slices with the adenovirus vector caused no significant increase in the induction of heat shock protein (HSP)-70 and showed no change in their electrophysiological properties as measured by stable field synaptic potentials in CA1 region and its reactivity to high frequency stimulation. Our data suggest that this adenovirus vector can be exploited to transfer multiple genes into neurons and may have implications for developing strategies for gene therapy. (C) 2000 Elsevier Science Inc. C1 Natl Inst Neurol Disorders & Stroke, Neurobiol Lab, NIH, Bethesda, MD 20892 USA. Natl Inst Neurol Disorders & Stroke, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Sihag, RK (reprint author), Natl Inst Neurol Disorders & Stroke, Neurobiol Lab, NIH, Bldg 36,Room 2A-21, Bethesda, MD 20892 USA. NR 28 TC 10 Z9 12 U1 1 U2 2 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0361-9230 J9 BRAIN RES BULL JI Brain Res. Bull. PD FEB PY 2000 VL 51 IS 3 BP 195 EP 202 DI 10.1016/S0361-9230(99)00217-8 PG 8 WC Neurosciences SC Neurosciences & Neurology GA 282HX UT WOS:000085213300001 PM 10718511 ER PT J AU Furey, ML Pietrini, P Alexander, GE Schapiro, MB Horwitz, B AF Furey, ML Pietrini, P Alexander, GE Schapiro, MB Horwitz, B TI Cholinergic enhancement improves performance on working memory by modulating the functional activity in distinct brain regions: A positron emission tomography regional cerebral blood flow study in healthy humans SO BRAIN RESEARCH BULLETIN LA English DT Article DE physostigmine; PET; rCBF; working memory; cognition ID ALZHEIMERS-DISEASE; BASAL FOREBRAIN; NEURAL SYSTEMS; PHYSOSTIGMINE; LESIONS; TASK; HIPPOCAMPUS; STIMULATION; SCOPOLAMINE; OBJECT AB Previously, we have shown that physostigmine, an acetylcholinesterase inhibitor, improved performance on a working memory for faces task, as reflected by reduced reaction time (RT), and reduced task-specific regional cerebral blood flow (rCBF) in right prefrontal cortex and, further, that these reductions in RT and right frontal rCBF were significantly correlated. Here we investigated the relation between the effects of physostigmine on task performance and task-specific functional brain response throughout the cortex by examining correlations between physostigmine-related changes in rCBF in all brain areas and changes in RT, In subjects who received an infusion of physostigmine, reduced RT correlated (p < 0.001) positively with reduced rCBF in right frontal cortex, left temporal cortex, anterior cingulate, and left hippocampus; and correlated with increased rCBF in medial occipital visual cortex, In subjects who received a placebo infusion of saline, no significant correlations between changes in RT and cortical rCBF were observed. The results show that cholinergically induced improvements in working memory performance are related to alterations in neural activity in multiple cortical regions, including increased neural activity in regions associated with early perceptual processing and decreased neural activity in regions associated with attention, memory encoding, and memory maintenance. (C) 2000 Elsevier Science Inc. C1 NIA, Neurosci Lab, NIH, Bethesda, MD USA. Univ Pisa, Dept Human & Environm Sci, Pisa, Italy. RP Furey, ML (reprint author), NIMH, Lab Brain & Cognit, NIH, Bldg 10,Rm 4C104,10 Ctr Dr,MSC 1366, Bethesda, MD 20892 USA. RI Furey, Maura/H-5273-2013 NR 35 TC 67 Z9 67 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0361-9230 J9 BRAIN RES BULL JI Brain Res. Bull. PD FEB PY 2000 VL 51 IS 3 BP 213 EP 218 DI 10.1016/S0361-9230(99)00219-1 PG 6 WC Neurosciences SC Neurosciences & Neurology GA 282HX UT WOS:000085213300003 PM 10718513 ER PT J AU Paradiso, A Simone, G Petroni, S Leone, B Vallejo, C Lacava, J Romero, A Machiavelli, M De Lena, M Allegra, CJ Johnston, PG AF Paradiso, A Simone, G Petroni, S Leone, B Vallejo, C Lacava, J Romero, A Machiavelli, M De Lena, M Allegra, CJ Johnston, PG TI Thymidilate synthase and p53 primary tumour expression as predictive factors for advanced colorectal cancer patients SO BRITISH JOURNAL OF CANCER LA English DT Article DE TS; p53; predictive factor; prognosis; colorectal cancer ID HUMAN THYMIDYLATE SYNTHASE; PROGNOSTIC-SIGNIFICANCE; SUPPRESSOR PROTEIN; CHEMOTHERAPY; GENE; LEUCOVORIN; CELLS; ADENOCARCINOMAS; 5-FLUOROURACIL; RESISTANCE AB The purpose of this work was to analyse the ability of p53 and thymidilate synthase (TS) primary tumour expression to retrospectively predict clinical response to chemotherapy and long-term prognosis in patients with advanced colorectal cancers homogeneously treated by methotrexate (MTX)-modulated-5-fluorouracil (5-FU-FA). A total of 108 advanced colorectal cancer patients entered the present retrospective study. Immunohistochemical p53 (pAb 1801 mAb) and TS (TS106 mAb) expression on formalin-fixed paraffin-embedded primary tumour specimens was related to probability of clinical response to chemotherapy, time to progression and overall survival. p53 was expressed in 53/108 (49%) tumours, while 54/108 (50%) showed TS immunostaining. No relationship was demonstrated between p53 positivity and clinical response to chemotherapy (objective response (OR): 20% vs 23%, in p53+ and 953- cases respectively) or overall survival. Percent of OR was significantly higher in TS-negative with respect to TS positive tumours (30% vs 15% respectively; P< 0.04); simultaneous analysis of TS and p53 indicated 7% OR for p53-positively-positive tumours vs 46% for p53-positive/TS-negative tumours (P < 0.03). Logistic regression analysis confirmed a significant association between TS tumour status and clinical response to chemotherapy (hazard ratio (HR): 2.91;95% confidence interval (CI) 8.34-1.01;two-sided P< 0.05). A multivariate analysis of overall survival showed that only a small number of metastatic sites was statistically relevant (HR 1.89; 95% CI 2.85-1.26; two-sided P < 0.03). Our study suggests that immunohistochemical expression of p53 and TS could assist the clinician in predicting response of colorectal cancer patients to modulated MTX5-FU therapy. (C) 2000 Cancer Research Campaign. C1 Inst Oncol, Clin Epidemiol Oncol Lab, I-70126 Bari, Italy. Inst Oncol, Histopathol Serv, I-70126 Bari, Italy. Grp Oncol Cooperat Sur, Neuquen, Argentina. Navy Med Oncol Branch, NIH, Bethesda, MD USA. Queens Univ Hosp, Dept Oncol, Belfast, Antrim, North Ireland. RP Paradiso, A (reprint author), Inst Oncol, Clin Epidemiol Oncol Lab, Via Amendola 209, I-70126 Bari, Italy. NR 42 TC 132 Z9 136 U1 0 U2 1 PU CHURCHILL LIVINGSTONE PI EDINBURGH PA JOURNAL PRODUCTION DEPT, ROBERT STEVENSON HOUSE, 1-3 BAXTERS PLACE, LEITH WALK, EDINBURGH EH1 3AF, MIDLOTHIAN, SCOTLAND SN 0007-0920 J9 BRIT J CANCER JI Br. J. Cancer PD FEB PY 2000 VL 82 IS 3 BP 560 EP 567 DI 10.1054/bjoc.1999.0964 PG 8 WC Oncology SC Oncology GA 279EP UT WOS:000085031700011 PM 10682666 ER PT J AU Yoo, JY Wang, XW Rishi, AK Lessor, T Xia, XM Gustafson, TA Hamburger, AW AF Yoo, JY Wang, XW Rishi, AK Lessor, T Xia, XM Gustafson, TA Hamburger, AW TI Interaction of the PA2G4 (EBP1) protein with ErbB-3 and regulation of this binding by heregulin SO BRITISH JOURNAL OF CANCER LA English DT Article DE ErbB-3; ErbB-2; heregulin; breast cancer ID EPIDERMAL GROWTH-FACTOR; MAMMARY EPITHELIAL-CELLS; PHOSPHATIDYLINOSITOL 3-KINASE; MOLECULAR-CLONING; GENE-PRODUCT; HUMAN-BREAST; CARCINOMA-CELLS; RECEPTOR; EXPRESSION; CDNA AB The processes by which ErbB-3, an inactive tyrosine kinase, exerts its biological effects are poorly understood. Using the yeast two-hybrid system, we have isolated an ErbB-3 binding protein (Ebp1) that interacts with the juxtamembrane domain of ErbB-3. This protein is identical to that predicted to be encoded for by the human PA2G4 gene. Ebp1 is the human homologue of a previously identified cell cycle-regulated mouse protein p38-2G4. Two transcripts of ebp1 mRNA (1.7 and 2.2 kb) were detected in several normal human organs. The interaction of Ebp1 with ErbB-3 was examined in vitro and in vivo. The first 15 amino acids of the juxtamembrane domain of ErbB-3 were essential for Ebp1 binding in vitro. Treatment of AU565 cells with the ErbB-3 ligand heregulin resulted in dissociation of Ebp1 from ErbB-3. Ebp1 translocated from the cytoplasm into the nucleus following heregulin stimulation. These findings suggest that Ebp1 may be a downstream member of an ErbB-3-regulated signal transduction pathway. (C) 2000 Cancer Research Campaign. C1 Univ Maryland, Mol & Cellular Biol Program, Baltimore, MD 21201 USA. Univ Maryland, Greenbaum Canc Ctr, Baltimore, MD 21201 USA. Univ Maryland, Dept Physiol, Baltimore, MD 21201 USA. Univ Maryland, Dept Pathol, Baltimore, MD 21201 USA. NCI, Human Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. RP Hamburger, AW (reprint author), Univ Maryland, Mol & Cellular Biol Program, 655 W Baltimore St, Baltimore, MD 21201 USA. RI Wang, Xin/B-6162-2009 FU NCI NIH HHS [F33CA63763, R01CA 76047] NR 37 TC 86 Z9 99 U1 0 U2 0 PU CHURCHILL LIVINGSTONE PI EDINBURGH PA JOURNAL PRODUCTION DEPT, ROBERT STEVENSON HOUSE, 1-3 BAXTERS PLACE, LEITH WALK, EDINBURGH EH1 3AF, MIDLOTHIAN, SCOTLAND SN 0007-0920 J9 BRIT J CANCER JI Br. J. Cancer PD FEB PY 2000 VL 82 IS 3 BP 683 EP 690 PG 8 WC Oncology SC Oncology GA 279EP UT WOS:000085031700028 PM 10682683 ER PT J AU Whitby, D Luppi, M Sabin, C Barozzi, P Di Biase, AR Balli, F Cucci, F Weiss, RA Boshoff, C Torelli, G AF Whitby, D Luppi, M Sabin, C Barozzi, P Di Biase, AR Balli, F Cucci, F Weiss, RA Boshoff, C Torelli, G TI Detection of antibodies to human herpesvirus 8 in Italian children: evidence for horizontal transmission SO BRITISH JOURNAL OF CANCER LA English DT Article ID SARCOMA-ASSOCIATED HERPESVIRUS; CLASSIC KAPOSIS-SARCOMA; HUMAN-HERPESVIRUS-8 INFECTION; SEXUAL TRANSMISSION; PREVALENCE; RISK; MEN; SEROPREVALENCE; CHILDHOOD; HISTORY AB Human herpesvirus 8 (HHV-8), also known as Kaposi's sarcoma associated herpesvirus (KSHV), has been shown to be the causative agent for Kaposi's sarcoma (KS) and to be more prevalent in populations or risk groups at increased risk for KS. HHV-8 infection is rare in children from the US and the UK, but has been reported in African children. In this study we examine HHV-8 infection in children from Italy, a country with an elevated prevalence of HHV-8 in adults and high socio-economic conditions. (C) 2000 Cancer Research Campaign. C1 Inst Canc Res, London SW3 6JB, England. Univ Modena, Policlin, Dept Med Sci, I-41100 Modena, Italy. Univ Modena, Policlin, Dept Paediat, Sect Haematol, I-41100 Modena, Italy. UCL Royal Free & Univ Coll Med Sch, Dept Primary Care & Populat Sci, London, England. Ctr Blood Transfus, Brindisi, Italy. RP Whitby, D (reprint author), NCI, Frederick Canc Res & Dev Ctr, POB B, Frederick, MD 21702 USA. RI Sabin, Caroline/C-2464-2008; Luppi, Mario/J-3668-2016; Barozzi, Patrizia/Q-2638-2016 OI Luppi, Mario/0000-0002-0373-1154; Barozzi, Patrizia/0000-0002-8936-1114 NR 27 TC 51 Z9 55 U1 0 U2 1 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0007-0920 J9 BRIT J CANCER JI Br. J. Cancer PD FEB PY 2000 VL 82 IS 3 BP 702 EP 704 PG 3 WC Oncology SC Oncology GA 279EP UT WOS:000085031700030 PM 10682685 ER PT J AU Hayes, RB Pottern, LM Strickler, H Rabkin, C Pope, V Swanson, GM Greenberg, RS Schoenberg, JB Liff, J Schwartz, AG Hoover, RN Fraumeni, JF AF Hayes, RB Pottern, LM Strickler, H Rabkin, C Pope, V Swanson, GM Greenberg, RS Schoenberg, JB Liff, J Schwartz, AG Hoover, RN Fraumeni, JF TI Sexual behaviour, STDs and risks for prostate cancer SO BRITISH JOURNAL OF CANCER LA English DT Article DE epidemiology; prostatic neoplasms; racial aspects; sexual behaviour ID HUMAN-PAPILLOMAVIRUS INFECTION; UNITED-STATES; EPIDEMIOLOGY; SYPHILIS; BLACKS; WHITES; CARCINOMA; HERPESVIRUS; CALIFORNIA; VASECTOMY AB A population-based case-control Study was carried out among 981 men (479 black, 502 white) with pathologically confirmed prostate cancer and 1315 controls (594 black, 721 white). In-person interviews elicited information on sexual behaviour and other potential risk factors for prostate cancer. Blood was drawn for serologic studies in a subset of the cases (n = 276) and controls (n = 295). Prostate cancer risk was increased among men who reported a history of gonorrhoea or syphilis (odds ratio (OR) = 1.6; 95% confidence internal (CI) 1.2-2.1) or showed serological evidence of syphilis (MHA-TP) (OR = 1.8; 95% CI 1.0-3.5). Patterns of risk for gonorrhoea and syphilis were similar for blacks (OR = 1.7; 95% CI 1.2-2.2) and whites (OR = 1.6; 95% CI 0.8-3.2). Risks increased with increasing occurrences of gonorrhoea, rising to OR = 3.3 (95% CI 1.4-7.8) among subjects with three or more events (P-trend = 0.0005). Frequent sexual encounters with prostitutes and failure to use condoms were also associated with increased risk. Syphilis, gonorrhoea, sex with prostitutes and unprotected sexual intercourse may be indicators of contact with a sexually transmissible factor that increases the risk of prostate cancer. (C) 2000 Cancer Research Campaign. C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. NIH, Off Director, Bethesda, MD USA. Ctr Dis Control & Prevent, Atlanta, GA USA. Michigan Canc Fdn, Detroit, MI 48201 USA. Emory Univ, Rollins Sch Publ Hlth, Dept Epidemiol, Atlanta, GA USA. RP Hayes, RB (reprint author), NCI, Div Canc Epidemiol & Genet, EPN 418, Bethesda, MD 20892 USA. OI Hayes, Richard/0000-0002-0918-661X FU NCI NIH HHS [N01-CN-51089, N01-CN-05225, N01-CP-5109] NR 51 TC 119 Z9 123 U1 4 U2 5 PU CHURCHILL LIVINGSTONE PI EDINBURGH PA JOURNAL PRODUCTION DEPT, ROBERT STEVENSON HOUSE, 1-3 BAXTERS PLACE, LEITH WALK, EDINBURGH EH1 3AF, MIDLOTHIAN, SCOTLAND SN 0007-0920 J9 BRIT J CANCER JI Br. J. Cancer PD FEB PY 2000 VL 82 IS 3 BP 718 EP 725 DI 10.1054/bjoc.1999.0986 PG 8 WC Oncology SC Oncology GA 279EP UT WOS:000085031700033 PM 10682688 ER PT J AU Yotsumoto, S Kanzaki, T Ko, MSH AF Yotsumoto, S Kanzaki, T Ko, MSH TI The beta subunit of the high-affinity IgE receptor, a candidate for atopic dermatitis, is not imprinted SO BRITISH JOURNAL OF DERMATOLOGY LA English DT Letter ID EPSILON-RI-BETA; IMMUNOGLOBULIN-E RECEPTOR; ASSOCIATION; EXPRESSION; VARIANT; MOUSE C1 Wayne State Univ, Sch Med, Ctr Mol Med & Genet, Detroit, MI 48202 USA. Kagoshima Univ, Fac Med, Dept Dermatol, Kagoshima 8908520, Japan. NIA, Dev Genom & Ageing Sect, Genet Lab, NIH, Baltimore, MD 21224 USA. RP Yotsumoto, S (reprint author), Wayne State Univ, Sch Med, Ctr Mol Med & Genet, 5047 Gullen Mall, Detroit, MI 48202 USA. RI Ko, Minoru/B-7969-2009 OI Ko, Minoru/0000-0002-3530-3015 NR 9 TC 3 Z9 3 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0007-0963 J9 BRIT J DERMATOL JI Br. J. Dermatol. PD FEB PY 2000 VL 142 IS 2 BP 370 EP 371 DI 10.1046/j.1365-2133.2000.03312.x PG 2 WC Dermatology SC Dermatology GA 291LX UT WOS:000085737300031 PM 10730778 ER PT J AU Bahceci, E Read, EJ Leitman, S Childs, R Dunbar, C Young, NS Barrett, AJ AF Bahceci, E Read, EJ Leitman, S Childs, R Dunbar, C Young, NS Barrett, AJ TI CD34+cell dose predicts relapse and survival after T-cell-depleted HLA-identical haematopoietic stem cell transplantation (HSCT) for haematological malignancies SO BRITISH JOURNAL OF HAEMATOLOGY LA English DT Article DE CD34+cells; allogeneic stem cell transplantation; haematological malignancy ID BONE-MARROW TRANSPLANTATION; VERSUS-HOST DISEASE; HEMATOLOGIC MALIGNANCIES; HIGH-RISK; LEUKEMIA; ENGRAFTMENT; CYTOMEGALOVIRUS; ADULTS; CSF AB Seventy-eight patients with haematological malignancies, received T-cell-depleted stem cell transplants and cyclosporin followed by delayed add-back, of donor lymphocytes to prevent leukaemia relapse. The source of stem cells was bone marrow in 50 patients and granulocyte colony-stimulating factor (G-CSF)-mobilized peripheral blood in 28 patients. In univariate analysis, only the CD34+ cell dose (but not the stem cell source or the T lymphocyte dose) and disease status were predictive for transplant-related mortality, relapse and survival. Patients receiving greater than or equal to 3 x 10(6) CD34+ cells/kg had an overall actuarial survival of 68% compared with 52%, 35% and 10%, respectively, for cell doses of 2-2.99, 1-1.99 and < 1 x 10(6)/kg, Multivariate analysis of risk factors for relapse identified disease risk and CD34+ cell dose as the only factors. Relapse was 62.5% in 38 patients at high risk of relapse vs, 25% for 40 patients at intermediate or low risk, CD34+ cell doses of greater than or equal to 3 x 10(6)/kg were associated with a 13.5% relapse vs. 48% for recipients of lower doses. This favourable effect of CD34+ cell dose on relapse was apparent in both high- and intermediate- plus low-risk groups. Our results support the potential benefit of a high stem cell dose in lowering transplant-related mortality (TRM) and in reducing relapse after allogeneic marrow or blood stem cell transplants. C1 NHLBI, Bone Marrow Transplant Unit, Hematol Branch, NIH, Bethesda, MD 20894 USA. NIH, Warren Grant Magnuson Clin Ctr, Dept Transfus Med, Bethesda, MD 20892 USA. RP Barrett, AJ (reprint author), NHLBI, Bone Marrow Transplant Unit, Hematol Branch, NIH, Bldg 10,Rm 7C103,9000 Rockville Pike, Bethesda, MD 20894 USA. NR 23 TC 55 Z9 56 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0007-1048 J9 BRIT J HAEMATOL JI Br. J. Haematol. PD FEB PY 2000 VL 108 IS 2 BP 408 EP 414 PG 7 WC Hematology SC Hematology GA 295MG UT WOS:000085970600031 PM 10691874 ER PT J AU Heinz, A Jones, DW AF Heinz, A Jones, DW TI Serotonin transporters in ecstasy users SO BRITISH JOURNAL OF PSYCHIATRY LA English DT Letter ID BETA-CIT BINDING; HUMAN BRAIN; IN-VIVO; DOPAMINE C1 NIMH, SPECT Lab, CBDB, NIH, Bethesda, MD 20792 USA. RP Heinz, A (reprint author), NIMH, SPECT Lab, CBDB, NIH, 10 Ctr Dr Rm 4C216, Bethesda, MD 20792 USA. NR 9 TC 17 Z9 17 U1 0 U2 0 PU ROYAL COLLEGE OF PSYCHIATRISTS PI LONDON PA BRITISH JOURNAL OF PSYCHIATRY 17 BELGRAVE SQUARE, LONDON SW1X 8PG, ENGLAND SN 0007-1250 J9 BRIT J PSYCHIAT JI Br. J. Psychiatry PD FEB PY 2000 VL 176 BP 193 EP 194 PG 2 WC Psychiatry SC Psychiatry GA 283TT UT WOS:000085292400019 PM 10755061 ER PT J AU Ratnasinghe, D Tangrea, JA Forman, MR Hartman, T Gunter, EW Qiao, YL Yao, SX Barett, MJ Giffen, CA Erozan, Y Tockman, MS Taylor, PR AF Ratnasinghe, D Tangrea, JA Forman, MR Hartman, T Gunter, EW Qiao, YL Yao, SX Barett, MJ Giffen, CA Erozan, Y Tockman, MS Taylor, PR TI Serum tocopherols, selenium and lung cancer risk among tin miners in China SO CANCER CAUSES & CONTROL LA English DT Article DE lung cancer; selenium; tocopherol; vitamin E ID VITAMIN-E; SUBSEQUENT RISK; BETA-CAROTENE; INTERVENTION TRIAL; FINNISH MEN; FOLLOW-UP; ANTIOXIDANTS; YOUNGER; COHORT; YUNNAN AB Objective: To evaluate the association of prediagnostic serum antioxidants and lung cancer risk we conducted a case-control study nested in an occupational cohort of tin miners. Methods: Male workers free of cancer enrolled in the cohort. During up to 6 years of follow-up, 339 lung cancer cases were diagnosed and, among these cases, those who donated blood prospectively (n = 108) were eligible for this study. For each case, two controls alive and free of cancer at the time of case diagnosis were matched on age and date of blood collection. Results: Overall, we observed no association between serum alpha-tocopherol, gamma-tocopherol or selenium levels and lung cancer risk. However, a significant gradient of decreasing lung cancer risk with increasing serum alpha-tocopherol was apparent for men less than 60 years old (odds ratio by tertile: 1.0, 0.9, 0.2; trend p = 0.002). Alpha-tocopherol was also protective in men who reported no alcohol drinking (OR by tertile: 1.0, 0.6, 0.3; trend p = 0.008). Conclusion: Although there were no significant overall associations between prospectively collected serum alpha-tocopherol, gamma-tocopherol or selenium and incidence of lung cancer, results from this study suggest that higher alpha-tocopherol levels may be protective in men less than 60 years old and in those who do not drink alcohol. C1 NCI, Div Clin Sci, NIH, Bethesda, MD USA. Ctr Dis Control & Prevent, NHANES Lab Biochem Anal, Atlanta, GA USA. Yunnan Tin Corp, Gejiu, Yunnan Province, Peoples R China. Informat Management Serv Inc, Silver Spring, MD USA. Johns Hopkins Univ, Sch Med, Dept Pathol, Baltimore, MD 21205 USA. H Lee Moffit Canc Ctr & Res Inst, Tampa, FL USA. RP Ratnasinghe, D (reprint author), NCI, DCS, Canc Prevent Studies Branch, 6006 Execut Blvd,Suite 321, Bethesda, MD 20892 USA. RI Qiao, You-Lin/B-4139-2012 OI Qiao, You-Lin/0000-0001-6380-0871 NR 32 TC 37 Z9 37 U1 1 U2 2 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0957-5243 J9 CANCER CAUSE CONTROL JI Cancer Causes Control PD FEB PY 2000 VL 11 IS 2 BP 129 EP 135 DI 10.1023/A:1008977320811 PG 7 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 275GE UT WOS:000084811800004 PM 10710196 ER PT J AU Dal Maso, L La Vecchia, C Franceschi, S Preston-Martin, S Ron, E Levi, F Mack, W Mark, SD McTiernan, A Kolonel, L Mabuchi, K Jin, F Wingren, G Galanti, MR Hallquist, A Glattre, E Lund, E Linos, D Negri, E AF Dal Maso, L La Vecchia, C Franceschi, S Preston-Martin, S Ron, E Levi, F Mack, W Mark, SD McTiernan, A Kolonel, L Mabuchi, K Jin, F Wingren, G Galanti, MR Hallquist, A Glattre, E Lund, E Linos, D Negri, E TI A pooled analysis of thyroid cancer studies. V. Anthropometric factors SO CANCER CAUSES & CONTROL LA English DT Article DE body mass index; case-control studies; height; risk; thyroid cancer; weight ID RISK; DISEASE; PAPILLARY; HISTORY; WEIGHT; HEIGHT; WOMEN; CARCINOMA; EXPOSURE; DIET AB Objective: To assess the relation between anthropometric factors and thyroid cancer risk in a pooled analysis of individual data from 12 case-control studies conducted in the US, Japan, China and Europe. Methods: 2056 female and 417 male cases, 3358 female and 965 male controls were considered. Odds ratios (OR) were derived from logistic regression, conditioning on age, A-bomb exposure (Japan) and study, and adjusting for radiotherapy. Results: Compared to the lowest tertile of height, the pooled OR was 1.2 for females for the highest one, and 1.5 for males, and trends in risk were significant. With reference to weight at diagnosis, the OR for females was 1.2 for the highest tertile, and the trend in risk was significant, whereas no association was observed in males. Body mass index (BMI) at diagnosis was directly related to thyroid cancer risk in females (OR = 1.2 for the highest tertile), but not in males. No consistent pattern of risk emerged with BMI during the late teens. Most of the associations were observed both for papillary and follicular cancers, and in all age groups. However, significant heterogeneity was observed across studies. Conclusions: Height and weight at diagnosis are moderately related to thyroid cancer risk. C1 Ist Ric Farmacol Mario Negri, Lab Gen Epidemiol, I-20157 Milan, Italy. Ctr Riferimento Oncol, I-33081 Aviano, PN, Italy. Univ Milan, Ist Stat Med & Biometria, I-20133 Milan, Italy. Univ So Calif, Dept Prevent Med, Los Angeles, CA 90033 USA. NCI, Div Canc Epidemiol & Genet, Rockville, MD 20852 USA. Registre Vaudois Tumeurs, CH-1011 Lausanne, Switzerland. Fred Hutchinson Canc Res Ctr, Seattle, WA 98109 USA. Univ Hawaii Manoa, Canc Res Ctr Hawaii, Honolulu, HI 96813 USA. Radiat Effects Res Fdn, Hiroshima 732, Japan. Shanghai Canc Inst, Shanghai, Peoples R China. Linkoping Univ, Fac Hlth Sci, Div Environm & Occupat Med, SE-58185 Linkoping, Sweden. Univ Uppsala Hosp, Dept Canc Epidemiol, SE-57185 Uppsala, Sweden. Karolinska Inst, Dept Oncol, SE-11235 Stockholm, Sweden. Stockholms Sjukhem, SE-11235 Stockholm, Sweden. Canc Registry Norway, NO-3010 Oslo, Norway. Univ Tromso, Inst Community Med, NO-9037 Tromso, Norway. Inst Prevent Med, GR-14561 Kifissia, Greece. RP Ron, E (reprint author), Ist Ric Farmacol Mario Negri, Lab Gen Epidemiol, Via Eritrea 62, I-20157 Milan, Italy. RI Negri, Eva/B-7244-2013 OI Negri, Eva/0000-0001-9712-8526 NR 28 TC 89 Z9 91 U1 0 U2 2 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0957-5243 J9 CANCER CAUSE CONTROL JI Cancer Causes Control PD FEB PY 2000 VL 11 IS 2 BP 137 EP 144 PG 8 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 275GE UT WOS:000084811800005 PM 10710197 ER PT J AU Mellemkjaer, L Johansen, C Gridley, G Linet, MS Kjaer, SK Olsen, JH AF Mellemkjaer, L Johansen, C Gridley, G Linet, MS Kjaer, SK Olsen, JH TI Crohn's disease and cancer risk (Denmark) SO CANCER CAUSES & CONTROL LA English DT Article DE cohort studies; Crohn's disease; inflammatory bowel diseases; intestinal neoplasms; risk ID INFLAMMATORY-BOWEL-DISEASE; ULCERATIVE-COLITIS; INTESTINAL CANCER; COLORECTAL-CANCER; POPULATION; MALIGNANCY AB Objectives: The large number of studies of intestinal cancer among patients with Crohn's disease have provided inconsistent risk estimates in regard to risk of both colorectal and small intestinal cancer. We investigated incidence of cancer among Crohn's disease patients in comparison with the incidence in the general population of Denmark. Methods: From the Danish National Registry of Patients we identified 2645 patients who had been hospitalized with Crohn's disease during 1977-1989. Cancer incidence for up to 17 years was determined in the cohort and compared to an expected number derived from national cancer incidence rates. Results: The 15 observed cases of colorectal cancer were close to the expected number of 13.1 (SIR = 1.1; 95% CI 0.6-1.9), whereas the five cases of small intestinal cancer (three adenocarcinomas and two carcinoids) observed corresponded to an 18-fold increased risk (SIR = 17.9; 95% CI 5.8-42). Conclusions: A potential excess of colorectal cancer among subgroups of patients with Crohn's disease was not detectable in the overall risk estimate for colorectal cancer. Only for small intestinal cancer was a significantly elevated risk found among these patients hospitalized with Crohn's disease. C1 Danish Canc Soc, Inst Canc Epidemiol, DK-2100 Copenhagen O, Denmark. NCI, Epidemiol & Biostat Program, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. RP Mellemkjaer, L (reprint author), Danish Canc Soc, Inst Canc Epidemiol, Strandblvd 49, DK-2100 Copenhagen O, Denmark. FU NCI NIH HHS [N01-CP-61111] NR 24 TC 59 Z9 59 U1 0 U2 2 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0957-5243 J9 CANCER CAUSE CONTROL JI Cancer Causes Control PD FEB PY 2000 VL 11 IS 2 BP 145 EP 150 DI 10.1023/A:1008988215904 PG 6 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 275GE UT WOS:000084811800006 PM 10710198 ER PT J AU Kelloff, GJ Sigman, CC Johnson, KM Boone, CW Greenwald, P Crowell, JA Hawk, ET Doody, LA AF Kelloff, GJ Sigman, CC Johnson, KM Boone, CW Greenwald, P Crowell, JA Hawk, ET Doody, LA TI Perspectives on surrogate end points in the development of drugs that reduce the risk of cancer SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Review ID CORONARY HEART-DISEASE; CERVICAL INTRAEPITHELIAL NEOPLASIA; PRIMARY-PREVENTION TRIAL; IN-SITU HYBRIDIZATION; BLADDER-CANCER; BARRETTS-ESOPHAGUS; GENETIC ALTERATIONS; CLINICAL-TRIALS; CLONAL ORIGIN; NECK-CANCER AB This paper proposes a scientific basis and possible strategy for applying surrogate end points in chemopreventive drug development. The potential surrogate end points for cancer incidence described are both phenotypic (at the tissue, cellular, and molecular levels) and genotypic biomarkers, To establish chemopreventive efficacy in randomized, placebo-controlled clinical trials, it is expected that in most cases it will be critical to ensure that virtually all of the biomarker lesions are prevented or that the lesions prevented are those with the potential to progress. This would require that both the phenotype and genotype of the target tissue in agent-treated subjects, especially in any new or remaining precancers, are equivalent to or show less progression than those of placebo-treated subjects. In the National Cancer Institute chemoprevention program, histological modulation of a precancer (intraepithelial neoplasia) has thus far been the primary phenotypic surrogate end point in chemoprevention trials. Additionally, we give high priority to biomarkers measuring specific and general genotypic changes correlating to the carcinogenesis progression model for the targeted cancer (e.g., progressive genomic instability as measured by loss of heterozygosity or amplification at a specific microsatellite loci). Other potential surrogate end points that may occur earlier in carcinogenesis are being analyzed in these precancers and in nearby normal appearing tissues. These biomarkers include proliferation and differentiation indices, specific gene and general chromosome damage, cell growth regulatory molecules, and biochemical activities (e.g., enzyme inhibition). Serum biomarkers also may be monitored (e.g., prostate-specific antigen) because of their accessibility, Potentially chemopreventive drug effects of the test agent also may be measured (e.g., tissue and serum estrogen levels in studies of steroid aromatase inhibitors). These initial studies are expected to expand the list of validated surrogate end points for future use. Continued discussion and research among the National Cancer Institute, the Food and Drug Administration, industry, and academia are needed to ensure that surrogate end point-based chemoprevention indications are feasible. C1 NCI, Div Canc Prevent, Bethesda, MD 20892 USA. CCS Associates, Mt View, CA 94043 USA. RP Kelloff, GJ (reprint author), NCI, Div Canc Prevent, EPN 201,MSC 7322,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 95 TC 59 Z9 63 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1055-9965 J9 CANCER EPIDEM BIOMAR JI Cancer Epidemiol. Biomarkers Prev. PD FEB PY 2000 VL 9 IS 2 BP 127 EP 137 PG 11 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 287FU UT WOS:000085496500001 PM 10698472 ER PT J AU Zheng, TZ Holford, TR Mayne, ST Tessari, J Ward, B Carter, D Owens, PH Boyle, P Dubrow, R Archibeque-Engle, S Dawood, O Zahm, SH AF Zheng, TZ Holford, TR Mayne, ST Tessari, J Ward, B Carter, D Owens, PH Boyle, P Dubrow, R Archibeque-Engle, S Dawood, O Zahm, SH TI Risk of female breast cancer associated with serum polychlorinated biphenyls and 1,1-dichloro-2,2 '-bis(p-chlorophenyl)ethylene SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Article ID ENVIRONMENTAL ORGANOCHLORINE EXPOSURE; BLOOD-LEVELS; ADIPOSE-TISSUE; DDT; WOMEN; RESIDUES; ESTROGENS; PESTICIDES; MEXICO; GROWTH AB This case-control study was designed to investigate the relationship between polychlorinated biphenyls (PCBs) and 1,1-dichloro-2,2'-bis(p-chlorophenyl)ethylene (DDE) and breast cancer risk in Connecticut. Cases were incident breast cancer patients who were either residents of Tolland County or who had a breast-related surgery at the Yale-New Haven Hospital in New Haven County. Controls were randomly selected from Tolland County residents or from patients who had newly diagnosed benign breast diseases or normal tissue at Yale-New Haven Hospital. A total of 475 cases and 502 controls had their serum samples analyzed for PCBs and DDE in 1995-1997, The age- and lipid-adjusted geometric mean serum level of DDE was comparable between the cases (460.1 ppb) and controls (456.2 ppb), The geometric mean serum level of PCBs was also comparable between cases (733.1 ppb) and controls (747.6 ppb), After adjustment for confounding factors, odds ratios of 0.96 (95% confidence interval, 0.67-1.36) for DDE and 0.95 (95% confidence interval, 0.68-1.32) for PCBs were observed when the third tertile was compared with the lowest. Further stratification by parity, lactation, and menopausal and estrogen receptor status also showed no significant association with serum levels of DDE or PCBs. The results by PCB congener groups also showed no major increased risk associated with any of the congener groups, Our study does not support the hypothesis that DDE and PCBs, as encountered through environmental exposure, increase the risk of female breast cancer. C1 Yale Univ, Sch Med, Dept Epidemiol, New Haven, CT 06510 USA. Yale Univ, Ctr Canc, New Haven, CT 06520 USA. Colorado State Univ, Ft Collins, CO 80523 USA. Ist Europeo Oncol, I-20141 Milan, Italy. NCI, Bethesda, MD 20892 USA. RP Zheng, TZ (reprint author), Yale Univ, Sch Med, Dept Epidemiol, 129 Church St,Suite 700-704, New Haven, CT 06510 USA. RI Boyle, Peter/A-4380-2014; Zahm, Shelia/B-5025-2015 OI Boyle, Peter/0000-0001-6251-0610; FU NCI NIH HHS [CA-62986] NR 43 TC 89 Z9 90 U1 1 U2 4 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1055-9965 J9 CANCER EPIDEM BIOMAR JI Cancer Epidemiol. Biomarkers Prev. PD FEB PY 2000 VL 9 IS 2 BP 167 EP 174 PG 8 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 287FU UT WOS:000085496500006 PM 10698477 ER PT J AU Olshan, AF Weissler, MC Watson, MA Bell, DA AF Olshan, AF Weissler, MC Watson, MA Bell, DA TI GSTM1, GSTT1, GSTP1, CYP1A1, and NAT1 polymorphisms, tobacco use, and the risk of head and neck cancer SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Article ID GLUTATHIONE-S-TRANSFERASE; ORAL-CANCER; SUSCEPTIBILITY; M1; GENOTYPES; LARYNX; LUNG; ACETYLTRANSFERASES; EPIDEMIOLOGY; POPULATION AB Squamous cell carcinoma of the head and neck (SCCHN), including the oral cavity, pharynx, and larynx, provides an ideal tumor model to investigate gene-environment interaction. We conducted a hospital-based case-control study including 182 cases with newly diagnosed SCCHN and 202 controls with nonneoplastic conditions of the head and neck that required surgery. Lifetime tobacco use and risk of SCCHN were evaluated in relation to the Polymorphisms Of GSTM1, GSTT1, GSTP1, CYP1A1, and NAT1. The main effects of genotype were associated with a slightly increased risk of SCCHN for GSTP1 [age-, race-, and sex-adjusted odds ratio (OR), 1.2; confidence interval (CI), 0.8-1.9], GSTT1 (OR, 1.2; CI, 0.7-2.3), and NAT1 (OR, 1.1; CI, 0.7-1.7), The joint effects of genotype combinations showed some excess risk for the combination of the GSTM1 null genotype and the CYP1A1 IIe/Val polymorphism (OR, 2.6; CI, 0.7-10.3). The analysis of the joint effects (interaction) of the "at-risk" genotypes and tobacco use did not reveal any interaction on either the multiplicative or additive scale for GSTM1, GSTP1, or CYP1A1, However, there was a suggestion of an interaction on the additive scale between the pack-years of tobacco use and the GSTT1 null genotype. The combined heterozygote and homozygote NAT1*10 genotypes also had a suggestive interaction with tobacco smoking history. The results of this study suggest a possible gene-environment interaction for certain carcinogen metabolizing enzymes, but larger studies that fully evaluate the interaction are needed. C1 Univ N Carolina, Sch Publ Hlth, Dept Epidemiol, Chapel Hill, NC 27599 USA. Univ N Carolina, Sch Med, Dept Surg, Div Otolaryngol Head & Neck Surg, Chapel Hill, NC 27599 USA. NIEHS, Lab Computat Biol & Risk Assessment, Res Triangle Pk, NC 27709 USA. RP Olshan, AF (reprint author), Univ N Carolina, Sch Publ Hlth, Dept Epidemiol, CB 7400, Chapel Hill, NC 27599 USA. FU NCI NIH HHS [CA61188] NR 33 TC 121 Z9 129 U1 0 U2 1 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1055-9965 J9 CANCER EPIDEM BIOMAR JI Cancer Epidemiol. Biomarkers Prev. PD FEB PY 2000 VL 9 IS 2 BP 185 EP 191 PG 7 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 287FU UT WOS:000085496500009 PM 10698480 ER PT J AU Baris, D Kwak, LW Rothman, N Wilson, W Manns, A Tarone, RE Hartge, P AF Baris, D Kwak, LW Rothman, N Wilson, W Manns, A Tarone, RE Hartge, P TI Blood levels of organochlorines before and after chemotherapy among non-Hodgkin's lymphoma patients SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Article ID POLYCHLORINATED BIPHENYL CONGENERS; CHLORINATED HYDROCARBONS; GAS-CHROMATOGRAPHY; BREAST-CANCER; RISK-FACTORS; TIME TRENDS; SERUM; PESTICIDES; TISSUE AB Several small studies suggest a link between environmental exposure to organochlorine compounds and risk of non Hodgkin's lymphoma (NHL). Because NHL is uncommon, studies of the topic often use a population-based case-control design, in which cases generally are enrolled after treatment has begun. If chemotherapy affects blood levels of organochlorines, exposure will be misclassified and findings distorted. To determine whether chemotherapy alters serum levels of organochlorines in NHL cases, we compared serum samples before and after treatment in 22 cases diagnosed with NHL between March 1994 and August 1995 and enrolled in a clinical trial at the United States National Cancer Institute's Clinical Center. The time difference between pretreatment and posttreatment samples ranged from 15 to 27 months with an average of 20 months. Laboratory analyses were conducted in blinded pretreatment and posttreatment pairs of the subjects. Pretreatment and posttreatment organochlorine serum levels were compared using Pearson correlation coefficient (r) and paired t test. The pretreatment and posttreatment serum levels were highly correlated for 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE) and polychlorinated biphenyls (PCBs) PCB-138, PCB-153, PCB-156, and total PCBs (ranging from 0.78 to 0.93), Serum levels of all of these organochlorines significantly decreased between initiation and completion of chemotherapy, 25% for total PCB (P = 0.0044), 28% for DDE (P = 0.0014), 25% for PCB-138 (P = 0.0053), 27% for PCB-153 (P = 0.0031), and 29% for PCB-156 (P = 0.045), Neither weight change nor lipid change was correlated with changes in chemical levels. There was no association between the length of time between blood draws and changes in chemical levels. Our data raise the possibility that lymphoma treatment depresses serum organochlorine levels. C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. NCI, Div Clin Sci, Bethesda, MD 20892 USA. RP Baris, D (reprint author), NCI, Div Canc Epidemiol & Genet, Execut Plaza S,Room 8122, Bethesda, MD 20892 USA. NR 21 TC 33 Z9 33 U1 0 U2 2 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1055-9965 J9 CANCER EPIDEM BIOMAR JI Cancer Epidemiol. Biomarkers Prev. PD FEB PY 2000 VL 9 IS 2 BP 193 EP 197 PG 5 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 287FU UT WOS:000085496500010 PM 10698481 ER PT J AU Whyatt, RM Perera, FP Jedrychowski, W Santella, RM Garte, S Bell, DA AF Whyatt, RM Perera, FP Jedrychowski, W Santella, RM Garte, S Bell, DA TI Association between polycyclic aromatic hydrocarbon-DNA adduct levels in maternal and newborn white blood cells and glutathione S-transferase P1 and CYP1A1 polymorphisms SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Article ID LUNG-CANCER; GENETIC POLYMORPHISMS; DIOL EPOXIDES; CATALYTIC EFFICIENCY; AIR-POLLUTION; SUSCEPTIBILITY; M1; GENOTYPES; VARIANTS; POLAND AB Polgcyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental pollutants; a number are carcinogenic. Metabolic polymorphisms may modulate susceptibility to PAM-induced DNA damage and carcinogenesis. This study investigates the relationship between PAH-DNA adduct levels (in maternal and newborn WBCs) and two polymorphisms: (a) an MspI RFLP in the 3' noncoding region of cytochrome P4501A1 (CYP1A1); and (b) an A-->G transition in nucleotide 313 of glutathione S-transferase PI (GSTP1), resulting in an ile105val substitution, CYP1A1 catalyzes the bioactivation of PAH; the CYP1A1 MspI RFLP has been associated with cancer of the lung, GSTP1 catalyzes the detoxification of PAH; the Pa1 allele has greater catalytic efficiency toward PAH diol epoxides, The study involves 160 mothers and their newborns from Poland. Regression models controlled for maternal smoking and other confounders. No association was seen between maternal adduct levels and either polymorphism, separately or combined, However, adduct levels were higher among newborns with the CYP1A1 MspI restriction site (heterozygotes and homozygotes combined) compared with newborns lacking the restriction site (P = 0.06), Adducts were higher among GSTP1 ile/val and ile/ile newborns compared with GSTP1 val/val newborns (P = 0.08), Adduct levels were 4-fold higher among GSTP1 ile/ile newborns having the CYP1A1 restriction site compared with GSTP1 val/val newborns who lacked the CYP1A1 restriction site (P = 0.04), This study demonstrates a significant combined effect of phase I and phase II polymorphisms on DNA damage from PAHs in fetal tissues. It illustrates the importance of considering interindividual variation in assessing risks of transplacental exposure to PAHs. C1 Columbia Univ, Joseph L Mailman Sch Publ Hlth, Div Environm Hlth Sci, New York, NY 10032 USA. Jagiellonian Univ, Coll Med, Krakow, Poland. Environm & Occupat Hlth Sci Inst, Piscataway, NJ 08854 USA. NIEHS, Res Triangle Pk, NC 27709 USA. RP Whyatt, RM (reprint author), Columbia Univ, Joseph L Mailman Sch Publ Hlth, Div Environm Hlth Sci, 60 Haven Ave,B-1, New York, NY 10032 USA. FU NCI NIH HHS [R01CA-39174, R01CA-35809]; NIEHS NIH HHS [1PO1-ESO5294, R01 ES008977, P01 ES009600] NR 60 TC 61 Z9 68 U1 0 U2 1 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1055-9965 J9 CANCER EPIDEM BIOMAR JI Cancer Epidemiol. Biomarkers Prev. PD FEB PY 2000 VL 9 IS 2 BP 207 EP 212 PG 6 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 287FU UT WOS:000085496500012 PM 10698483 ER PT J AU Hursting, S Shen, J Klein, R Guan, Y Chang, S Wei, Q AF Hursting, S Shen, J Klein, R Guan, Y Chang, S Wei, Q TI Low-dose genistein induces cyclin-dependent kinase inhibitors and G1 cell cycle arrest in prostate cancer cells. SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1055-9965 J9 CANCER EPIDEM BIOMAR JI Cancer Epidemiol. Biomarkers Prev. PD FEB PY 2000 VL 9 IS 2 BP 234 EP 234 PG 1 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 287FU UT WOS:000085496500023 ER PT J AU Stolzenberg-Solomon, RZ Pietinen, P Barrett, MJ Taylor, PR Virtamo, J Albanes, D AF Stolzenberg-Solomon, RZ Pietinen, P Barrett, MJ Taylor, PR Virtamo, J Albanes, D TI Dietary folate and pancreas cancer risk in a prospective cohort of male smokers SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Meeting Abstract C1 NCI, Canc Prevent Studies Branch, Bethesda, MD 20892 USA. RI Albanes, Demetrius/B-9749-2015 NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1055-9965 J9 CANCER EPIDEM BIOMAR JI Cancer Epidemiol. Biomarkers Prev. PD FEB PY 2000 VL 9 IS 2 BP 234 EP 234 PG 1 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 287FU UT WOS:000085496500025 ER PT J AU Daly, T Royal, RE Kershaw, MH Treisman, J Wang, G Li, WP Herlyn, D Eshhar, Z Hwu, P AF Daly, T Royal, RE Kershaw, MH Treisman, J Wang, G Li, WP Herlyn, D Eshhar, Z Hwu, P TI Recognition of human colon cancer by T cells transduced with a chimeric receptor gene SO CANCER GENE THERAPY LA English DT Article DE immunotherapy; colon cancer; lymphocyte; gene therapy; redirection ID CARCINOMA-ASSOCIATED ANTIGEN; HIGH-DOSE INTERLEUKIN-2; ACTIVATION MOTIF ITAM; MONOCLONAL-ANTIBODIES; COLORECTAL-CARCINOMA; GASTROINTESTINAL CARCINOMA; GAMMA-CHAIN; TUMOR-CELLS; LYMPHOCYTES; GROWTH AB Transduction with chimeric T-cell receptor genes can be used to redirect primary T lymphocytes to recognize specific antigens (Ags), including, ovarian and breast cancer Ags. To extend this approach to colon cancer we report here redirection of T cells using a chimeric receptor recognizing the colon cancer-associated Ag EGP40. Chimeric T cell receptors were constructed by ligating single-chain genes of either of two EGP40-specific monoclonal antibodies (CO17.1A, GA733) to the Fe receptor gamma-signaling chain. Retroviral vectors incorporating these constructs were used to transduce a murine T-cell line and human peripheral blood lymphocytes. These modified T cells were analyzed for specific recognition of colon cancer lines by measuring cytokine release and lyric activity against tumor targets. Murine lymphocytes transduced with the chimeric receptor based on GA733, but not CO17.1A, released cytokine specifically in response to EGP40-expressing colon cancer cell lines. Recognition of colon cancer targets by murine lymphocytes was blocked by the addition of GA733 antibody or soluble EGP40 Ag, confirming that colon cancer recognition is based on specific chimeric receptor-Ag interaction. Human lymphocytes transduced with chimeric GA733 specifically recognized colon carcinoma cells in cytokine release assays and lysed EGP40-expressing tumor cells. Genetic modification of T cells can be used to redirect T cells against EGP40-expressing tumor cells. The expression of chimeric GA733 in the autologous lymphocytes of patients may provide a source of tumor-reactive cells with therapeutic application against colon cancer. C1 NCI, NIH, Surg Branch, Tumor Immunol Sect, Bethesda, MD 20892 USA. Univ Penn, Sch Med, Wistar Inst Anat & Biol, Philadelphia, PA 19104 USA. Weizmann Inst Sci, IL-76100 Rehovot, Israel. RP Hwu, P (reprint author), NCI, NIH, Surg Branch, Tumor Immunol Sect, Bldg 10,Room 2B42,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 47 TC 41 Z9 43 U1 1 U2 2 PU STOCKTON PRESS PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 0929-1903 J9 CANCER GENE THER JI Cancer Gene Ther. PD FEB PY 2000 VL 7 IS 2 BP 284 EP 291 DI 10.1038/sj.cgt.7700121 PG 8 WC Biotechnology & Applied Microbiology; Oncology; Genetics & Heredity; Medicine, Research & Experimental SC Biotechnology & Applied Microbiology; Oncology; Genetics & Heredity; Research & Experimental Medicine GA 299BR UT WOS:000086176400016 PM 10770638 ER PT J AU Paul, D Qazilbash, MH Song, KM Xu, H Sinha, BK Liu, J Cowan, KH AF Paul, D Qazilbash, MH Song, KM Xu, H Sinha, BK Liu, J Cowan, KH TI Construction of a recombinant adeno-associated virus (rAAV) vector expressing murine interleukin-12 (IL-12) SO CANCER GENE THERAPY LA English DT Article DE adeno-associated virus; interleukin-12 vector; gene transfer ID CYTOKINE GENE-THERAPY; T RESPONSES INVITRO; WILD-TYPE P53; ADENOASSOCIATED VIRUS; IN-VIVO; ANTITUMOR IMMUNITY; HUMAN-LYMPHOCYTES; TUMOR-REGRESSION; COLON-CARCINOMA; NK CELLS AB IL-12 is a heterodimeric cytokine that is known to induce tumor regression and long-term antitumor immunity. Recombinant adeno-associated virus (rAAV) vectors are advantageous for gene therapy in that they lack pathogenicity in humans, infect dividing as well as nondividing cells, and show a broad range of infectivity. We constructed an rAAV vector expressing interleukin-12 (IL-12) for cancer immunotherapy studies in a mouse model by inserting murine IL-12 (mIL-12) p35 and p40 cDNAs into the plasmid pRep4 and inserting the encephalomyocarditis virus internal ribosomal entry site between the p35 and p40 cDNAs. The mIL-12 expression cassette containing the Rous sarcoma virus promoter and a simian virus 40 polyadenylation signal was subcloned into the AAV plasmid p008Sub/NeoR which contains two AAV inverted terminal repeat sequences and the NeoR gene driven by the thymidine kinase promoter. rAAV virions (10(4) infectious particles/ml) were generated by cotransfection of rAAV-mIL-12 and a helper plasmid (pAAV/Ad) into 293 cells previously infected with adenovirus 5. After infection of DG fibroblasts with rAAV-mIL-12, G418-resistant clones were isolated. Each of the 1D D6 clones isolated produced up to 5.2 ng/10(6) cells/48 hours of mIL-12 as determined by enzyme-linked immunosorbent assay. Induction of interferon-gamma, enhanced lymphocyte proliferation, and cytotoxicity assays confirmed biologically functional IL-12 production by the vector. This is the First report indicating that an rAAV vector expresses mIL-12, which can be used to model the effects of mIL-12 alone and/or in combination with Other antitumor agents. C1 NCI, NIH, Med Branch, Bethesda, MD 20892 USA. NEI, NIH, Bethesda, MD 20892 USA. NHLBI, NIH, Hematol Branch, Bethesda, MD 20892 USA. RP Cowan, KH (reprint author), NCI, NIH, Med Branch, 10-12N-226, Bethesda, MD 20892 USA. NR 59 TC 11 Z9 12 U1 0 U2 3 PU STOCKTON PRESS PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 0929-1903 J9 CANCER GENE THER JI Cancer Gene Ther. PD FEB PY 2000 VL 7 IS 2 BP 308 EP 315 DI 10.1038/sj.cgt.7700105 PG 8 WC Biotechnology & Applied Microbiology; Oncology; Genetics & Heredity; Medicine, Research & Experimental SC Biotechnology & Applied Microbiology; Oncology; Genetics & Heredity; Research & Experimental Medicine GA 299BR UT WOS:000086176400019 PM 10770641 ER PT J AU Khaled, HM Aly, MS Magrath, IT AF Khaled, HM Aly, MS Magrath, IT TI Loss of Y chromosome in bilharzial bladder cancer SO CANCER GENETICS AND CYTOGENETICS LA English DT Article ID INSITU HYBRIDIZATION; INTERPHASE CYTOGENETICS; PROSTATIC ADENOCARCINOMA; DNA PROBES; TUMORS; FISH; ABERRATIONS; ANEUPLOIDY; CELLS; SET AB Bilharzial bladder cancer is the most common malignant neoplasm in Egypt, also occurring with a high incidence in other regions of the Middle East and East Africa. In a previous study, using centromere probes specific for chromosomes 3, 4, 7-11, 16, and 17, we demonstrated that monosomy of chromosome 9 (48.4%), and numerical aberrations of chromosome 17 (19.4%) were the most common observed imbalances. The present study extends the establishment of the baseline cytogenetic profile of this type of malignancy. Interphase cytogenetics by fluorescence in situ hybridization with the use of a panel of centromere-associated DNA probes for chromosomes 1, 2, 5, 6, 12, 13/21, 14, 15, 18, 19, 20, X, and Y was performed on paraffin-embedded bladder specimens from 25 Egyptian patients affected with bilharzial bladder cancer. No numerical aberrations were detected in the 25 cases for chromosomes 1, 2, 5, 6, 12, 13/21, 14, is, 18, 19, 20, and X. However, loss of chromosome Y was observed in 7 of the 17 male cases studied (41.2%). No significant correlation was observed between loss of the Y chromosome and any of the different clinicopathologic characteristics of these cases. These data suggest that loss of the Y chromosome is the second frequent event that can occur in bilharzial bladder cancer. A molecular genetic model of bilharzial bladder cancer is evolving. (C) Elsevier Science Inc., 2000. All rights reserved. C1 Cairo Univ, Natl Canc Inst, Dept Med Oncol, Cairo, Egypt. Cairo Univ, Fac Sci, Beni Suef, Egypt. NCI, Pediat Branch, NIH, Bethesda, MD 20892 USA. RP Aly, MS (reprint author), POB 169 Imbaba, Cairo 12411, Egypt. NR 24 TC 12 Z9 12 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0165-4608 J9 CANCER GENET CYTOGEN JI Cancer Genet. Cytogenet. PD FEB PY 2000 VL 117 IS 1 BP 32 EP 36 DI 10.1016/S0165-4608(99)00126-0 PG 5 WC Oncology; Genetics & Heredity SC Oncology; Genetics & Heredity GA 284PH UT WOS:000085340500006 PM 10700863 ER PT J AU Atkins, MB Kunkel, L Sznol, M Rosenberg, SA AF Atkins, MB Kunkel, L Sznol, M Rosenberg, SA TI High-dose recombinant interleukin-2 therapy in patients with metastatic melanoma: Long-term survival update SO CANCER JOURNAL FROM SCIENTIFIC AMERICAN LA English DT Article; Proceedings Paper CT 2nd International Symposium on the Use of Interleukin-2 for the Treatment of Cancer CY JUL 29-31, 1999 CL SAN FRANCISCO, CALIFORNIA DE immunotherapy; interleukin-2; melanoma ID RENAL-CELL CARCINOMA; PHASE-I TRIAL; MALIGNANT-MELANOMA; RANDOMIZED TRIAL; COMBINATION; CANCER; RECEPTOR; CHEMOTHERAPY; PREVENTION; CISPLATIN AB PURPOSE To update response duration and survival data for patients with metastatic melanoma receiving the high-dose TV bolus recombinant interleukin (IL)-2 regimen. PATIENTS AND METHODS Two hundred seventy assessable patients were entered into eight clinical trials conducted between 1985 and 1993. IL-2 600,000 or 720,000 IU/kg was administered by 15-minute intravenous infusion every 8 hours for up to 14 consecutive doses over 5 days as clinically tolerated with maximum support, including pressors. A second, identical cycle of treatment was scheduled following 6 to 9 days of rest, and courses could be repeated every 6 to 12 weeks in stable or responding patients. Responding patients received up to five courses (two cycles/course) of treatment. All data were updated through December 1998 using report forms completed by the clinical investigators. RESULTS The objective overall response rate was unchanged from the previous report. Tumor responses were seen in 16% of patients, with complete responses in 17 (6%) and partial responses in 26 (10%). Median survival for the group as a whole is now 12 months. Median follow-up time for surviving patients exceeds 7 years. Median duration of response for the 43 responding patients and the 26 patients with partial responses remained unchanged at 8.9 and 5.9 months, respectively. Response durations ranged from 1.5 to > 122 months. The median duration of complete responses has yet to be reached, but is at least 59 months. Thirty-one patients (11%) were alive as of last contact; 28 were confirmed, including 18 responding patients. Three patients were lost to follow-up at > 1, > 13, and > 104 months. Twelve responding patients remained continually disease- or progression-free from > 70 to > 150 months following initiation of therapy. Disease progression was not observed in any patient who was responding as of the last report or in any patient responding for longer than 30 months. CONCLUSION These data continue to support the notion that high-dose IL-2 produces durable responses in some patients with metastatic melanoma and should be considered a therapeutic option for appropriately selected patients with this disease. C1 Harvard Univ, Beth Israel Deaconess Med Ctr, Sch Med, Div Hematol Oncol,Cytokine Working Grp, Boston, MA 02115 USA. CHIRON, Emeryville, CA USA. Natl Canc Inst, Bethesda, MD USA. RP Atkins, MB (reprint author), Harvard Univ, Beth Israel Deaconess Med Ctr, Sch Med, Div Hematol Oncol,Cytokine Working Grp, 330 Brookline Ave E-KS-158, Boston, MA 02115 USA. NR 33 TC 282 Z9 286 U1 2 U2 9 PU SCI AMERICAN INC PI NEW YORK PA 415 MADISON AVE, NEW YORK, NY 10017 USA SN 1081-4442 J9 CANCER J SCI AM JI Cancer J. Sci. Am. PD FEB PY 2000 VL 6 SU 1 BP S11 EP S14 PG 4 WC Oncology SC Oncology GA 283PL UT WOS:000085283900004 PM 10685652 ER PT J AU Fisher, RI Rosenberg, SA Fyfe, G AF Fisher, RI Rosenberg, SA Fyfe, G TI Long-term survival update for high-dose recombinant interleukin-2 in patients with renal cell carcinoma SO CANCER JOURNAL FROM SCIENTIFIC AMERICAN LA English DT Article; Proceedings Paper CT 2nd International Symposium on the Use of Interleukin-2 for the Treatment of Cancer CY JUL 29-31, 1999 CL SAN FRANCISCO, CALIFORNIA DE immunotherapy; interleukin-2; renal cell carcinoma ID ACTIVATED KILLER-CELLS; CONTINUOUS INFUSION INTERLEUKIN-2; PHASE-II TRIAL; THERAPY; CANCER; BOLUS AB PURPOSE To update response duration and survival data for patients with metastatic renal cell carcinoma treated with high-dose interleukin (IL)-2. PATIENTS AND METHODS Two hundred fifty-five assessable patients were entered onto seven phase II clinical trials. Recombinant IL-2 600,000 or 720,000 IU/kg was administered by 15-minute intravenous infusion every 8 hours for up to 14 consecutive doses over 5 days as clinically tolerated with maximal support. A second, identical cycle of treatment was scheduled following 5 to 9 days of rest, and courses could be repeated every 6 to 12 weeks in stable or responding patients. All data were updated as of December 1998, with report forms completed by the clinical investigators. These data had last been updated as part of the Food and Drug Administration reporting requirements in 1996. RESULTS Objective responses previously have been reported in 37 of 255 patients (15%) with 17 complete responses (7%) and 20 partial responses (8%). These data remain unchanged from previous reports. Median response duration for all objective responders remains unchanged at 54 months, but the range now extends from 3 to > 131 months. Median duration for all complete responses has not yet been reached, but was at least 80 months (range, 7- > 131 mo) at the time of this analysis. Median duration for all partial responses remains 20 months (range, > 126 mo). Median survival time for all 255 patients remains 16.3 months, with 10% to 20% of patients estimated to be alive 5 to 10 years after treatment with high-dose IL-2. CONCLUSION With prolonged follow-up, treatment with high-dose recombinant IL-2 remains extremely effective for a subset of patients with metastatic renal cell carcinoma. C1 Loyola Univ, Med Ctr, Cardinal Bernardin Canc Ctr, Maywood, IL 60153 USA. Natl Canc Inst, Bethesda, MD USA. CHI RON, Emeryville, CA USA. RP Fisher, RI (reprint author), Loyola Univ, Med Ctr, Cardinal Bernardin Canc Ctr, Room 255,, Maywood, IL 60153 USA. NR 11 TC 273 Z9 276 U1 0 U2 1 PU SCI AMERICAN INC PI NEW YORK PA 415 MADISON AVE, NEW YORK, NY 10017 USA SN 1081-4442 J9 CANCER J SCI AM JI Cancer J. Sci. Am. PD FEB PY 2000 VL 6 SU 1 BP S55 EP S57 PG 3 WC Oncology SC Oncology GA 283PL UT WOS:000085283900012 PM 10685660 ER PT J AU Overwijk, WW Theoret, MR Restifo, NP AF Overwijk, WW Theoret, MR Restifo, NP TI The future of interleukin-2: Enhancing therapeutic anticancer vaccines SO CANCER JOURNAL FROM SCIENTIFIC AMERICAN LA English DT Article; Proceedings Paper CT 2nd International Symposium on the Use of Interleukin-2 for the Treatment of Cancer CY JUL 29-31, 1999 CL SAN FRANCISCO, CALIFORNIA DE adjuvant; cancer vaccines; gp100; interleukin-2; melanoma; T lymphocytes ID TUMOR-INFILTRATING LYMPHOCYTES; ESTABLISHED PULMONARY METASTASES; DOSE RECOMBINANT INTERLEUKIN-2; ACTIVATION-INDUCED DEATH; HUMAN T-CELLS; IMMUNE-RESPONSES; MICE; ANTIGEN; CANCER; IMMUNIZATION AB PURPOSE The purpose of our efforts is to trigger the immune destruction of established cancer. Interleukin (IL)-2 can mediate the regression of tumors in patients with melanoma and renal cell carcinoma In animal models, the antitumor effects of IL-2 are mediated by T lymphocytes. Stimulation with specific antigen can enhance the ability of T cells to respond to IL-2 by triggering the rapid upregulation of the high-affinity IL-2 receptor. We are seeking to design recombinant and synthetic vaccines capable of preferentially priming T cells with specificity for tumor cells. METHODS The antitumor activity of experimental vaccines is being studied preclinically using recently developed murine models that employ the mouse homologues of human tumor-associated antigens, Once the most effective experimental vaccines are optimized in experimental animals, clinical trials can be conducted. Vaccines are being evaluated for their ability to mediate the regression of established tumors, and a variety of immunologic correlates are being measured RESULTS In animal models, vaccines based on molecularly defined tumor-associated antigens expressed in viral vectors or delivered as "naked" DNA stimulate the expansion of CD4+ and CD8+ tumor-specific T lymphocytes, Coadministration of IL-2 with these vaccines dramatically enhances their ability to mediate the regression of established cancer, In the clinic, treatment of melanoma patients with peptide vaccine and IL-2 resulted in objective responses in approximately 40% of patients, a response rate more than twice that typically achieved with IL-2 alone. Paradoxically, tumor-specific CD8+ T-cell levels were nor increased in these patients. CONCLUSION The addition of recombinant and synthetic cancer vaccines to a regimen of IL-2 can result in improved antitumor responses in both animal models and melanoma patients. Vaccine-primed, tumor-specific T cells may preferentially proliferate upon administration of IL-2. The apparent lack of increase in CD8+ T-cell numbers in this setting suggests that the vaccine-primed T cells functionally disappear after a transient period of activation, Preventing the disappearance of activated T cells upon IL-2 administration-for example, by blocking proapoptotic signals-may enhance the therapeutic effectiveness of anticancer vaccines. C1 NCI, Surg Branch, NIH, Bethesda, MD 20892 USA. Howard Hughes Med Inst, Natl Inst Hlth, Res Scholars Program, Bethesda, MD 20817 USA. RP Overwijk, WW (reprint author), NCI, Surg Branch, NIH, Bldg 10,Room 2B42, Bethesda, MD 20892 USA. RI Restifo, Nicholas/A-5713-2008 FU Intramural NIH HHS [Z01 BC010763-01, NIH0010139353, Z99 TW999999] NR 36 TC 50 Z9 52 U1 0 U2 1 PU SCI AMERICAN INC PI NEW YORK PA 415 MADISON AVE, NEW YORK, NY 10017 USA SN 1081-4442 J9 CANCER J SCI AM JI Cancer J. Sci. Am. PD FEB PY 2000 VL 6 SU 1 BP S76 EP S80 PG 5 WC Oncology SC Oncology GA 283PL UT WOS:000085283900016 PM 10685664 ER PT J AU Rosenberg, SA AF Rosenberg, SA TI Interleukin-2 and the development of immunotherapy for the treatment of patients with cancer SO CANCER JOURNAL FROM SCIENTIFIC AMERICAN LA English DT Article; Proceedings Paper CT 2nd International Symposium on the Use of Interleukin-2 for the Treatment of Cancer CY JUL 29-31, 1999 CL SAN FRANCISCO, CALIFORNIA ID TUMOR-INFILTRATING LYMPHOCYTES; CYTOLYTIC T-LYMPHOCYTES; METASTATIC CANCER; HLA-A2 MELANOMAS; ANTIGEN; GENE; IDENTIFICATION; TYROSINASE; REGRESSION; PEPTIDES C1 NCI, Div Clin Sci, Surg Branch, NIH, Bethesda, MD 20892 USA. RP Rosenberg, SA (reprint author), NCI, Div Clin Sci, Surg Branch, NIH, Bldg 10,Room 2B42,10 Ctr Dr,MSC 1502, Bethesda, MD 20892 USA. NR 36 TC 40 Z9 42 U1 1 U2 2 PU SCI AMERICAN INC PI NEW YORK PA 415 MADISON AVE, NEW YORK, NY 10017 USA SN 1081-4442 J9 CANCER J SCI AM JI Cancer J. Sci. Am. PD FEB PY 2000 VL 6 SU 1 BP S2 EP S7 PG 6 WC Oncology SC Oncology GA 283PL UT WOS:000085283900002 PM 10685650 ER PT J AU Wattenberg, LW Wiedmann, TS Estensen, RD Zimmerman, CL Galbraith, AR Steele, VE Kelloff, GJ AF Wattenberg, LW Wiedmann, TS Estensen, RD Zimmerman, CL Galbraith, AR Steele, VE Kelloff, GJ TI Chemoprevention of pulmonary carcinogenesis by brief exposures to aerosolized budesonide or beclomethasone dipropionate and by the combination of aerosolized budesonide and dietary myo-inositol SO CARCINOGENESIS LA English DT Article ID FEMALE A/J MICE; BENZOPYRENE-INDUCED NEOPLASIA; DEXAMETHASONE; MYOINOSITOL; INHIBITION; LUNG AB This investigation is part of an effort to develop chemoprevention for carcinogenesis of the lung. It focuses on the efficacy of low doses of synthetic glucocorticoids administered either as single agents or in combination with a second compound, myo-inositol, Glucocorticoids are potent inhibitors of carcinogenesis. The use of low doses is important to avoid potential side-effects. The synthetic glucocorticoid budesonide, administered by aerosol for 20 a three times a week, was studied to determine its effects on benzo[a]pyrene-induced pulmonary adenoma formation in female A/J mice. Two dose levels were employed, 10 and 25 mu g/kg body wt. The lower dose produced a 34% reduction in lung tumor formation and the higher dose level a 60% reduction in lung tumors. In additional groups of mice, the effects of 0.3% myo-inositol added to the diet was found to reduce pulmonary tumor formation by 53%. The two agents given in combination resulted in a greater inhibition of lung tumor formation than either by itself. Budesonide at 10 mu g/kg body wt plus 0.3% myo-inositol reduced the number of tumors by 60% and budesonide at 25 mu g/kg body wt plus 0.3% inyo-inositol reduced lung tumor formation by 79%. To determine whether a glucocorticoid other than budesonide would have inhibitory effects in this experimental model, beclomethasone dipropionate administered by aerosol for 20 s three times a week was studied as a single agent and showed almost identical inhibitory properties to budesonide, The doses of the glucocorticoids calculated on a daily basis are within the range of those used widely for control of chronic allergic respiratory diseases in the human. The capacity of low doses of inhaled glucocorticoids to prevent pulmonary neoplasia and the enhancement of this preventive effect by myo-inositol, an essentially non-toxic compound, are findings that should encourage further work to evaluate the applicability of these agents to the prevention of neoplasia of the lung in the human. C1 Univ Minnesota, Sch Pharm, Dept Lab Med & Pathol, Minneapolis, MN 55455 USA. Univ Minnesota, Coll Pharmaceut, Sch Pharm, Minneapolis, MN 55455 USA. NCI, Bethesda, MD 20892 USA. RP Wattenberg, LW (reprint author), Univ Minnesota, Sch Pharm, Dept Lab Med & Pathol, Minneapolis, MN 55455 USA. FU NCI NIH HHS [N01-CN-75114] NR 20 TC 97 Z9 97 U1 0 U2 1 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0143-3334 J9 CARCINOGENESIS JI Carcinogenesis PD FEB PY 2000 VL 21 IS 2 BP 179 EP 182 DI 10.1093/carcin/21.2.179 PG 4 WC Oncology SC Oncology GA 287JU UT WOS:000085503400009 PM 10657955 ER PT J AU Shilkaitis, A Green, A Steele, V Lubet, R Kelloff, G Christov, K AF Shilkaitis, A Green, A Steele, V Lubet, R Kelloff, G Christov, K TI Neoplastic transformation of mammary epithelial cells in rats is associated with decreased apoptotic cell death SO CARCINOGENESIS LA English DT Article ID PRENEOPLASTIC LESIONS; HORMONE DEPENDENCE; BREAST-CANCER; GLAND; CARCINOGENESIS; PROLIFERATION; KINETICS; TUMORS; MICE; 7,12-DIMETHYLBENZANTHRACENE AB Previous studies have shown that terminal end buds (TEBs) in the murine mammary gland have high proliferative activity and demonstrate apoptotic cell death (ACD), Since TEBs are considered the place of origin of most chemically induced mammary carcinomas, we hypothesized that the development of hyperplastic and premalignant (carcinoma in situ, CIS) lesions in TEBs is associated with either a further increase in cell proliferation and/or with a decrease in ACD, To test this hypothesis we used the N-methyl-N-nitorosourea (MNU) carcinogenesis model in rats, where the occurrence of mammary tumors is preceded by hyperplastic and premalignant lesions arising mostly in TEBs, as well as in ducts and alveoli. The percentage of proliferating cells, as evaluated by 5-bromodeoxyuridine labeling (BrdU-LI), was similar in TEBs to those in terminal endbud hyperplasia (TEBH), CIS, and carcinomas (CA), whereas the percentage of apoptotic cells (apoptotic index, AI) was relatively high in TEBs and decreased in TEBH, CIS, and CA. This indicates that neoplastic transformation of mammary epithelial cells in TEBs is not associated with an increase in cell proliferation, but with a decrease in ACD, In addition to TEBH, hyperplastic lesions developed in ductal branching areas (ductal hyperplasia, DH) and alveolar structures (alveolar hyperplasia, AH). However, BrdU-LI in both DH and AH was lower than in TEBH, whereas the AI values were similar, suggesting that TEBH has a higher potential for progression and malignant transformation than DH and AH. In mammary tumors apoptotic cells were rare in the peripheral, proliferative areas, but frequent close to the necrotic areas, suggesting that intratumoral factors may significantly affect ACD, Thus, it appears that dissociation between cell proliferation and apoptosis occurs in the hyperplastic stages of mammary carcinogenesis and that neoplastic transformation of mammary epithelial cells is associated with decreased ACD but not with increased cell proliferation. C1 Univ Illinois, Dept Surg Oncol, Chicago, IL 60612 USA. Natl Canc Inst, Div Chemoprevent, Bethesda, MD USA. RP Christov, K (reprint author), Univ Illinois, Dept Surg Oncol, 840 S Wood St M-C, Chicago, IL 60612 USA. FU NCI NIH HHS [N01-CN-55179-MAO, N01-CN-65031-MAO] NR 35 TC 20 Z9 20 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0143-3334 J9 CARCINOGENESIS JI Carcinogenesis PD FEB PY 2000 VL 21 IS 2 BP 227 EP 233 DI 10.1093/carcin/21.2.227 PG 7 WC Oncology SC Oncology GA 287JU UT WOS:000085503400016 PM 10657962 ER PT J AU Souici, AC Mirkovitch, J Hausel, P Keefer, LK Felley-Bosco, E AF Souici, AC Mirkovitch, J Hausel, P Keefer, LK Felley-Bosco, E TI Transition mutation in codon 248 of the p53 tumor suppressor gene induced by reactive oxygen species and a nitric oxide-releasing compound SO CARCINOGENESIS LA English DT Article ID DNA STRAND BREAKAGE; HYDROGEN-PEROXIDE TREATMENT; CELLULAR-ENERGY DEPLETION; PEROXYNITRITE IN-VITRO; LIPID-PEROXIDATION; OXIDATIVE DAMAGE; BACTERIAL LIPOPOLYSACCHARIDE; TERT-BUTYLHYDROPEROXIDE; BASE MODIFICATION; EPITHELIAL-CELLS AB Exposing the human bronchial epithelial cell line BEAS2B to the nitric oxide (NO) donor sodium 1-(N,N-diethylamino)diazen-1-ium-1,2-diolate (DEA/NO) at an initial concentration of 0.6 mM while generating superoxide ion at the rate of 1 mu M/min with the hypoxanthine/xanthine oxidase (HX/XO) system induced C:G-->T:A transition mutations in codon 248 of the p53 gene. This pattern of mutagenicity was not seen by 'fish-restriction fragment length polymorphism/polymerase chain reaction' (fish-RFLP/PCR) on exposure to DEA/NO alone, however, exposure to HX/XO led to various mutations, suggesting that co-generation of NO and superoxide was responsible for inducing the observed point mutation. DEA/NO potentiated the ability of HX/XO to induce lipid peroxidation as well as DNA single- and double-strand breaks under these conditions, while 0.6 mM DEA/NO in the absence of HX/XO had no significant effect on these parameters. The results show that a point mutation seen at high frequency in certain common human tumors can be induced by simultaneous exposure to reactive oxygen species and a NO source. C1 Univ Lausanne, Inst Pharmacol & Toxicol, CH-1005 Lausanne, Switzerland. Swiss Inst Expt Canc Res, CH-1066 Epalinges, Switzerland. NCI, Frederick Canc Res & Dev Ctr, Comparat Carcinogenesis Lab, Chem Sect, Ft Detrick, MD 21702 USA. RP Felley-Bosco, E (reprint author), Univ Lausanne, Inst Pharmacol & Toxicol, Rue Bugnon 27, CH-1005 Lausanne, Switzerland. RI Keefer, Larry/N-3247-2014; Felley-Bosco, Emanuela/E-7484-2017 OI Keefer, Larry/0000-0001-7489-9555; Felley-Bosco, Emanuela/0000-0002-3408-0294 NR 86 TC 36 Z9 42 U1 0 U2 2 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0143-3334 J9 CARCINOGENESIS JI Carcinogenesis PD FEB PY 2000 VL 21 IS 2 BP 281 EP 287 DI 10.1093/carcin/21.2.281 PG 7 WC Oncology SC Oncology GA 287JU UT WOS:000085503400023 PM 10657969 ER PT J AU Rashid, H Marshall, RJ Diver, DJ Breall, JA AF Rashid, H Marshall, RJ Diver, DJ Breall, JA TI Spontaneous and diffuse coronary artery spasm unresponsive to conventional intracoronary pharmacological therapy: A case report SO CATHETERIZATION AND CARDIOVASCULAR INTERVENTIONS LA English DT Article DE spasm; myocardial infarction; nitrates; calcium blockers ID VARIANT ANGINA; MYOCARDIAL-INFARCTION; STENOSIS; PATHOGENESIS; ANGIOPLASTY; VASOSPASM; WOMEN AB We report a case of coronary artery spasm in a woman without atherosclerotic risk factors. Coronary arteries were initially normal angiographically; however, repeat angiogram for severe chest pain revealed diffuse spasm of left circumflex system. Vasospasm did not respond to intracoronary pharmacotherapy and resulted in Q-wave myocardial infarction. Published 2000 Wiley-Liss, Inc.(dagger) C1 NHLBI, Clin Trials Sci Res Grp, Div Epidemiol & Clin Applicat, Bethesda, MD 20892 USA. Georgetown Univ, Med Ctr, Div Cardiol, Washington, DC 20007 USA. Georgetown Univ, Med Ctr, Inst Cardiovasc Sci, Washington, DC 20007 USA. RP Rashid, H (reprint author), NHLBI, Clin Trials Sci Res Grp, Div Epidemiol & Clin Applicat, Rockledge Ctr 2,6701 Rockledge Dr,Room 8149, Bethesda, MD 20892 USA. NR 25 TC 3 Z9 3 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 1522-1946 J9 CATHETER CARDIO INTE JI Catheter. Cardiovasc. Interv. PD FEB PY 2000 VL 49 IS 2 BP 188 EP 191 DI 10.1002/(SICI)1522-726X(200002)49:2<188::AID-CCD16>3.0.CO;2-6 PG 4 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 278VG UT WOS:000085009200016 PM 10642771 ER PT J AU Griffiths, EJ Ocampo, CJ Savage, JS Stern, MD Silverman, HS AF Griffiths, EJ Ocampo, CJ Savage, JS Stern, MD Silverman, HS TI Protective effects of low and high doses of cyclosporin A against reoxygenation injury in isolated rat cardiomyocytes are associated with differential effects on mitochondrial calcium levels SO CELL CALCIUM LA English DT Article ID CYTOSOLIC FREE CALCIUM; REPERFUSION INJURY; PERMEABILITY TRANSITION; ISCHEMIA-REPERFUSION; CARDIAC MYOCYTES; OXIDATIVE STRESS; BRAIN-DAMAGE; CA2+; HEPATOCYTES; HEARTS AB In this study we aimed to determine the concentration range of cyclosporin A (CsA) which was effective in protecting against reoxygenation injury in isolated cardiomyocytes, and its effects on intramitochondrial free calcium levels ([Ca2+](m)). We also determined whether a high [CsA] had any deleterious effect on normal myocyte function. Isolated adult rat ventricular myocytes were placed in a chamber on the stage of a fluorescence microscope for induction of hypoxia, [Ca2+](m) was determined from indo-1/am loaded cells where the cytosolic fluorescence signal had been quenched by superfusion with Mn2+. Cell length was measured using an edge-tracking device. Upon induction of hypoxia, control cells underwent rigor-contracture in 37 +/- 1 min (n=99) (T1); CsA had no effect on T1. The percentage of control cells which recovered upon reoxygenation depended on the time spent in rigor (T2), With a T2 of 21-30 min, only 36% of control cells recovered compared with 90% and 78% of cells treated with 0.2 mu M and 1 mu M CsA respectively. After 40 min in rigor, [Ca2+](m) was 280+/-60 nM in control-recovered cells (50% of cells) and 543+/-172 nM and 153+/-26 nM in cells treated with 0.2 and 1 mu M CsA, respectively (all CsA treated cells recovered). in normoxic studies, CsA had no effect on cell contractility or [Ca2+](m) upon rapid pacing, even in presence of an elevated external [Ca2+]. In conclusion, both low and high [CsA] protected against reoxygenation injury to cardiomyocytes despite having opposing effects on [Ca2+](m), suggesting more than one mechanism of action. CsA had no effect on either cell contractility or [Ca2+](m) in normoxic cells. (C) 2000 Harcourt Publishers Ltd. C1 Bristol Royal Infirm, Bristol Heart Inst, Bristol BS2 8HW, Avon, England. Johns Hopkins Univ Hosp, Div Cardiol, Baltimore, MD 21205 USA. NIH, Gerontol Res Ctr, Cardiovasc Sci Lab, Baltimore, MD USA. RP Griffiths, EJ (reprint author), Bristol Royal Infirm, Bristol Heart Inst, Level 7, Bristol BS2 8HW, Avon, England. NR 46 TC 46 Z9 51 U1 0 U2 1 PU CHURCHILL LIVINGSTONE PI EDINBURGH PA JOURNAL PRODUCTION DEPT, ROBERT STEVENSON HOUSE, 1-3 BAXTERS PLACE, LEITH WALK, EDINBURGH EH1 3AF, MIDLOTHIAN, SCOTLAND SN 0143-4160 J9 CELL CALCIUM JI Cell Calcium PD FEB PY 2000 VL 27 IS 2 BP 87 EP 95 DI 10.1054/ceca.1999.0094 PG 9 WC Cell Biology SC Cell Biology GA 294RM UT WOS:000085924200003 PM 10756975 ER PT J AU De Santis, ML Martinez-Lacaci, I Bianco, C Seno, M Wallace-Jones, B Kim, N Ebert, A Wechselberger, C Salomon, DS AF De Santis, ML Martinez-Lacaci, I Bianco, C Seno, M Wallace-Jones, B Kim, N Ebert, A Wechselberger, C Salomon, DS TI Cripto-1 induces apoptosis in HC-11 mouse mammary epithelial cells SO CELL DEATH AND DIFFERENTIATION LA English DT Article DE apoptosis; Cripto-1; caspase activation; mammary epithelial cells ID GROWTH-FACTOR-ALPHA; BREAST-CANCER CELLS; GLAND INVOLUTION; TRANSGENIC MICE; DOWN-REGULATION; EXPRESSION; DIFFERENTIATION; AMPHIREGULIN; SURVIVAL; DEATH AB Cripto-1 (CR-1) is an epidermal growth factor (EGF)-related protein. CR-1 can inhibit beta-casein and whey acidic protein expression in mouse mammary epithelial cells. The present study demonstrates that CR-1 can induce apoptosis in HC-11 mouse mammary epithelial cells, as measured by bisbenzimide stained nuclei, TUNEL assay and cell death ELISA, Apoptosis could be observed after 2 days of exposure of confluent HC-11 cells to CR-1 in the absence of the survival factors EGF and insulin, with maximum apoptosis occurring at 3 days. A reduction in poly(ADP-ribose) polymerase (PARP) expression and an increase in beta-catenin cleavage was found after 18 h of exposure to CR-1 suggesting that apoptosis was preceded by the induction of a caspase activity since the caspase inhibitor ZFAD.FMK could block the CR-1-induced reduction in PARP expression and CR-1-induced apoptosis, CR-1 was found to increase the expression of caspase-3-like protease, Although, the levels of p27(kip1) and p21Bax did not change after exposure to CR-1 for 18 h, the levels of Bcl-x(L) became undetectable. These studies suggest that CR-1 promotes apoptosis by mediating the induction of caspase-3-like protease and downregulating the expression of Bcl-x(L). C1 NCI, Tumor Growth Factor Sect, Tumor Immunol & Biol Lab, NIH, Bethesda, MD 20892 USA. Walter Reed Army Med Ctr, Walter Reed Army Inst Res, Dept Mol Pathol, Washington, DC 20307 USA. Okayama Univ, Fac Engn, Dept Biosci & Biotechnol, Okayama 7008530, Japan. Free Univ Berlin, Dept Obstet, D-1000 Berlin, Germany. Free Univ Berlin, Dept Gynecol, D-1000 Berlin, Germany. RP Salomon, DS (reprint author), NCI, Tumor Growth Factor Sect, Tumor Immunol & Biol Lab, NIH, Bldg 10,Room 5B39, Bethesda, MD 20892 USA. RI SENO, Masaharu /B-2092-2011 OI SENO, Masaharu /0000-0001-8547-6259 NR 51 TC 11 Z9 11 U1 0 U2 1 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 1350-9047 J9 CELL DEATH DIFFER JI Cell Death Differ. PD FEB PY 2000 VL 7 IS 2 BP 189 EP 196 DI 10.1038/sj.cdd.4400588 PG 8 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 292VU UT WOS:000085816600006 PM 10713733 ER PT J AU Preston, GA Srinivasan, D Barrett, JC AF Preston, GA Srinivasan, D Barrett, JC TI Apoptotic response to growth factor deprivation involves cooperative interactions between c-Fos and p300 SO CELL DEATH AND DIFFERENTIATION LA English DT Article DE p300; c-Fos; apoptosis; p53; transcription factor ID CREB-BINDING-PROTEIN; ADENOVIRUS E1A PROTEINS; PROGRAMMED CELL-DEATH; TUMOR-SUPPRESSOR GENE; HAMSTER EMBRYO CELLS; NF-KAPPA-B; TRANSCRIPTIONAL ACTIVATION; NEOPLASTIC PROGRESSION; COACTIVATOR P300; IN-VITRO AB Two preneoplastic cell lines have been utilized to study changes in the regulation of apoptosis during neoplastic progression [sup(+)I (stage I) and sup(-)II (stage II)]. Sup(+)I cells are prone to undergo apoptosis, while sup(-)II cells are relatively resistant. We report that induction of apoptosis in sup(+)I cells is tightly correlated with the formation of c-Fos/p300 complexes, which were not present in the non-apoptotic sup(-)II cells under the same conditions. When apoptosis was induced in the sup(-)II cells by over-expression of c-Fos, concomitant c-Fos:p300 complexes were detected. Overexpression of p300 resulted in apoptosis in sup(-)II cells and also in p53(wt) human tumor cells, but not in p53(mutant) human tumor cells. Over-expression of the C-terminal fragment of p300, which contains the c-fos binding site, enhanced apoptosis, suggesting that the c-fos:p300 complex is actively involved in apoptosis. We propose that p300 could function as a general mediator of transcription factor-induced apoptosis. C1 NIEHS, Mol Carcinogenesis Lab, NIH, Res Triangle Pk, NC 27709 USA. Univ N Carolina, Dept Med, Div Nephrol, Chapel Hill, NC 27599 USA. Harvard Univ, Massachusetts Gen Hosp, Ctr Canc, Grad Program Biol & Biomed Sci, Charlestown, MA 02129 USA. RP Preston, GA (reprint author), Univ N Carolina, Div Nephrol & Hypertens, CB 7155,346 MacNider Bld, Chapel Hill, NC 27599 USA. NR 70 TC 19 Z9 20 U1 0 U2 0 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 1350-9047 J9 CELL DEATH DIFFER JI Cell Death Differ. PD FEB PY 2000 VL 7 IS 2 BP 215 EP 226 DI 10.1038/sj.cdd.4400637 PG 12 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 292VU UT WOS:000085816600009 PM 10713736 ER PT J AU Holden, PR Hasmall, SC James, NH West, DR Brindle, RD Gonzalez, FJ Peters, JM Roberts, RA AF Holden, PR Hasmall, SC James, NH West, DR Brindle, RD Gonzalez, FJ Peters, JM Roberts, RA TI Tumour necrosis factor alpha (TNF alpha): Role in suppression of apoptosis by the peroxisome proliferator nafenopin SO CELLULAR AND MOLECULAR BIOLOGY LA English DT Article DE TNF alpha; peroxisome proliferator activated receptor alpha (PPAR alpha); apoptosis; DNA synthesis; non-genotoxic carcinogenesis ID NF-KAPPA-B; ACTIVATED RECEPTOR; DNA-SYNTHESIS; CELL-DEATH; PPAR-ALPHA; RAT-LIVER; KUPFFER CELLS; ALTERED FOCI; NONGENOTOXIC HEPATOCARCINOGEN; PHENOTYPIC-EXPRESSION AB The peroxisome proliferator (PPs) class of non-genotoxic rodent hepatocarcinogens induce mouse hepatocytes DNA synthesis and suppress apoptosis, This phenotype can be reproduced in vitro using exogenous tumour necrosis factor alpha (TNF alpha), suggesting a role for TNF alpha in mediating the liver growth response to PPs. In hepatocytes isolated from the peroxisome proliferator activated receptor alpha (PPAR alpha) null mouse, PPs are unable to stimulate DNA synthesis or to suppress either spontaneous or TCF beta 1-induced apoptosis. However, the ability of TNF alpha to modulate hepatocyte survival and growth is unaltered, suggesting that TNF alpha acts independently or downstream of PPAR alpha to mediate the growth changes associated with PPAR alpha activation. Since PPAR alpha is a ligand activated transcription factor, we determined if TNF alpha gene expression was altered by PP treatment during an early time window preceding PP-induced growth changes. However there was no induction of TNF alpha expression by nafenopin over the constitutive levels noted in control cultured cells. In summary, TNF alpha acts downstream or independently of PPAR alpha to mediate the suppression of apoptosis and induction of DNA synthesis by PPs. In this in vitro model, the PP nafenopin do nor appear to mediate de novo TNF alpha gene expression suggesting that the response to nafenopin may be mediated by bioactivation or release of pre-existing TNF alpha protein from Kupffer cells. C1 Zeneca Cent Toxicol Lab, Macclesfield SK10 4TJ, Cheshire, England. NCI, Mol Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. RP Roberts, RA (reprint author), Zeneca Cent Toxicol Lab, Alderley Pk, Macclesfield SK10 4TJ, Cheshire, England. RI Peters, Jeffrey/D-8847-2011 NR 57 TC 19 Z9 22 U1 0 U2 0 PU CELLULAR & MOLECULAR BIOLOGY PI NOISY-LE-GRAND PA PROF R WEGMANN RESIDENCE HAUSSMANN 1 AVENUE DU PAVE NEUF, 93160 NOISY-LE-GRAND, FRANCE SN 0145-5680 J9 CELL MOL BIOL JI Cell. Mol. Biol. PD FEB PY 2000 VL 46 IS 1 BP 29 EP 39 PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 289GL UT WOS:000085612100004 PM 10726969 ER PT J AU Ganpat, MM Nishimura, M Toyoshige, M Okuya, S Pointer, RH Rebois, RV AF Ganpat, MM Nishimura, M Toyoshige, M Okuya, S Pointer, RH Rebois, RV TI Evidence for stimulation of adenylyl cyclase by an activated G(s) heterotrimer in cell membranes - An experimental method for controlling the G(s) subunit composition of cell membranes SO CELLULAR SIGNALLING LA English DT Article DE stimulatory heterotrimeric G protein; subunit dissociation; adenylyl cyclase; cholera toxin; cell membranes ID G-PROTEINS; REGULATORY COMPONENT; LIPID MODIFICATIONS; BINDING PROTEINS; LYMPHOMA-CELLS; ALPHA-SUBUNIT; GTP-BINDING; DISSOCIATION; RECEPTOR; FLUOROALUMINATE AB Heterotrimeric (alpha beta gamma) G(s) mediates agonist-induced stimulation of adenylyl cyclase (AC). Cholera toxin (CTx) will ADP-ribosylate the alpha-subunit of G(s) (G(s)alpha). G(s)alpha-deficient cyc(-) membranes were "stripped" of GP. When the stripped cyc(-) were incubated with G(s)alpha and/or G beta gamma, each was incorporated into the membranes independently of the other. Both G(s)alpha and G beta gamma had to be present in the membranes, and they had to be able to form a heterotrimer in order for CTx to ADP-ribosylate G(s)alpha, indicating that the membrane bound G(s) heterotrimer is a substrate for CTx, but the G(s)alpha subunit by itself is not. When G(s)alpha was completely and irreversibly activated with GTP gamma S and incorporated into stripped cyc(-), it was a poor substrate for CTx and a weak stimulator of AC unless G beta gamma was also incorporated. Furthermore, the level of AC stimulation corresponded to the amount of G(s) heterotrimer that was formed in the membranes from GTP gamma S-activated G(s)alpha and G beta gamma. These data suggest that AC is stimulated by an activated G(s) heterotrimer in cell membranes. (C) 2000 Elsevier Science Inc. All rights reserved. C1 NINDS, Membrane Biochem Sect, Lab Mol & Cellular Neurobiol, NIH, Bethesda, MD 20892 USA. RP Rebois, RV (reprint author), NINDS, Membrane Biochem Sect, Lab Mol & Cellular Neurobiol, NIH, Bldg 49,Room 2A28, Bethesda, MD 20892 USA. EM rebois@box-r.nih.gov NR 30 TC 14 Z9 14 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0898-6568 J9 CELL SIGNAL JI Cell. Signal. PD FEB PY 2000 VL 12 IS 2 BP 113 EP 122 DI 10.1016/S0898-6568(99)00078-9 PG 10 WC Cell Biology SC Cell Biology GA 285UR UT WOS:000085407000006 PM 10679580 ER PT J AU Werhahn, KJ Wiseman, K Herzog, J Forderreuther, S Dichgans, M Straube, A AF Werhahn, KJ Wiseman, K Herzog, J Forderreuther, S Dichgans, M Straube, A TI Motor cortex excitability in patients with migraine with aura and hemiplegic migraine SO CEPHALALGIA LA English DT Article DE transcranial magnetic stimulation; motor cortex; cortical excitability; migraine; familial hemiplegic migraine ID TRANSCRANIAL MAGNETIC STIMULATION; INTERICTAL CORTICAL EXCITABILITY; HUMANS; INHIBITION; HYPEREXCITABILITY; BRAIN; POTENTIALS; NEOCORTEX; RESPONSES; GENE AB We studied the excitability of the motor cortex using transcranial magnetic stimulation (TMS) in 12 patients with migraine with aura (MA) and nine patients with familial hemiplegic migraine (FHM). Motor thresholds at rest, the duration of the cortical and peripheral silent period and intracortical inhibition and facilitation using paired-pulse TMS at intervals of 2 to 15 ms were measured with patients free of attacks for at least 48 h. In contrast to previous reports we could not find any significant differences between patient groups and compared to controls (n=17) in the parameters tested. The results suggest that there are no interictal changes of excitability of the motor cortex in migraine. This study does not support the concept of general cortical hyperexcitability in migraine secondary to a genetic predisposition or a structural alteration of inhibitory interneurones in the cortex due to repeated parenchymal insults during attacks. C1 Univ Munich, Dept Neurol, D-8000 Munich, Germany. RP Werhahn, KJ (reprint author), NINDS, NIH, Human Cort Physiol Sect, Bld 10,Room 5N242,10 Ctr Dr, Bethesda, MD 20892 USA. NR 34 TC 53 Z9 53 U1 0 U2 1 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0333-1024 J9 CEPHALALGIA JI Cephalalgia PD FEB PY 2000 VL 20 IS 1 BP 45 EP 50 DI 10.1046/j.1468-2982.2000.00011.x PG 6 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 315DP UT WOS:000087097600007 PM 10817446 ER PT J AU Chaffin, CL Trewin, AL Hutz, RJ AF Chaffin, CL Trewin, AL Hutz, RJ TI Estrous cycle-dependent changes in the expression of aromatic hydrocarbon receptor (AHR) and AHR-nuclear translocator (ARNT) mRNAs in the rat ovary and liver SO CHEMICO-BIOLOGICAL INTERACTIONS LA English DT Article DE aromatic hydrocarbon receptor; aromatic hydogen receptor nuclear translocator protein; ovary; liver; estrous cycle; estrogen ID IN-UTERO; 2,3,7,8-TETRACHLORODIBENZO-P-DIOXIN TCDD; GRANULOSA-CELLS; HOLTZMAN RATS; MODULATION; SECRETION; MECHANISM; PROTEIN AB The aromatic hydrocarbon receptor (AHR) and AHR nuclear translocator protein (ARNT) mediate the toxic effects of a wide variety of halogenated and polycyclic aromatic hydrocarbons. While it can be assumed that AHR has an endogenous function, its role in reproduction is currently undefined. The present study seeks to examine the regulation of AHR and ARNT mRNAs in liver and ovarian tissues across the rat estrous cycle. Message for hepatic AHR was increased significantly on the morning of proestrus, and decreased dramatically by the evening of proestrus; while hepatic ARNT mRNA was significantly decreased between diestrus and the morning of proestrus, and between the evening of proestrus and the morning of estrus. Ovarian AHR mRNA was unchanged from diestrus to proestrus, and was decreased on the evening of proestrus. Changes in the expression of ARNT mRNA mirrored changes in the liver. To assess interaction between the AHR- and estrogen-receptor (ER)-signaling pathways and to test the hypothesis that estrogen regulates AHR mRNA, 25-day-old female rats were injected with either 17 beta-estradiol, the ER antagonist ICI 182 780, or with vehicle, and hepatic AHR mRNA was measured. Treatment with estrogen or the estrogen antagonist did not alter the abundance of AHR mRNA in the liver. These data suggest that while estrogen may not be the key regulator of AHR mRNA expression, a factor associated with the rat reproductive cycle may be important in regulating the expression of both the AHR and ARNT genes in the ovary and liver. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved. C1 Univ Wisconsin, Dept Biol Sci, Milwaukee, WI 53211 USA. UWM, NIEHS Marine & Freshwater Biomed MFB Sci Ctr, Milwaukee, WI 53211 USA. RP Hutz, RJ (reprint author), Univ Wisconsin, Dept Biol Sci, 3209 N Maryland Ave, Milwaukee, WI 53211 USA. FU NIEHS NIH HHS [ES08342, ES06807] NR 30 TC 32 Z9 32 U1 0 U2 1 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0009-2797 J9 CHEM-BIOL INTERACT JI Chem.-Biol. Interact. PD FEB 1 PY 2000 VL 124 IS 3 BP 205 EP 216 DI 10.1016/S0009-2797(99)00157-X PG 12 WC Biochemistry & Molecular Biology; Pharmacology & Pharmacy; Toxicology SC Biochemistry & Molecular Biology; Pharmacology & Pharmacy; Toxicology GA 284VN UT WOS:000085353700005 PM 10728779 ER PT J AU Hurd, S AF Hurd, S TI The impact of COPD on lung health worldwide - Epidemiology and incidence SO CHEST LA English DT Article DE COPD; education programs; morbidity; public health AB Information on the prevalence of COPD was obtained from vital statistics, health interview surveys, hospital charge records, national publications, and the World Health Organization (WHO). These data indicate that COPD is a common disease with implications for global health. In the United States, morbidity caused by COPD is 4%, making COPD the fourth leading cause of death, exceeded only by heart attacks, cancer, and stroke. Internationally, there is substantial variation in death rates possibly reflecting smoking behavior, type and processing of tobacco, pollution, climate, respiratory management, and genetic factors. The Global Obstructive Lung Disease Initiative, initiated by the National Heart, Lung, and Blood Institute and the WHO, aims to raise awareness of the increasing burden of COPD, decrease morbidity and mortality, promote further study of the condition, and implement programs to, prevent COPD. C1 NHLBI, Div Lung Dis, Bethesda, MD 20892 USA. RP Hurd, S (reprint author), NHLBI, Div Lung Dis, 6701 Rockledge Dr,Suite 10122, Bethesda, MD 20892 USA. NR 2 TC 90 Z9 92 U1 2 U2 6 PU AMER COLL CHEST PHYSICIANS PI NORTHBROOK PA 3300 DUNDEE ROAD, NORTHBROOK, IL 60062-2348 USA SN 0012-3692 J9 CHEST JI Chest PD FEB PY 2000 VL 117 IS 2 SU S BP 1 EP 4 PG 4 WC Critical Care Medicine; Respiratory System SC General & Internal Medicine; Respiratory System GA 287WU UT WOS:000085530300001 ER PT J AU Reischl, IG AF Reischl, IG TI Dendritic cells and lung antigen responses SO CLINICAL AND EXPERIMENTAL ALLERGY LA English DT Editorial Material ID COLONY-STIMULATING FACTOR; IN-VITRO; HEMATOPOIETIC PROGENITORS; PERIPHERAL-BLOOD; T-CELLS; MONOCYTES; DIFFERENTIATION; MACROPHAGES; MATURATION; GENERATION C1 Southampton Gen Hosp, Southampton SO16 6YD, Hants, England. RP Reischl, IG (reprint author), NIH, 9000 Rockville Pike,Bldg 10,9N256, Bethesda, MD 20892 USA. NR 28 TC 2 Z9 2 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0954-7894 J9 CLIN EXP ALLERGY JI Clin. Exp. Allergy PD FEB PY 2000 VL 30 IS 2 BP 160 EP 163 PG 4 WC Allergy; Immunology SC Allergy; Immunology GA 287NF UT WOS:000085511600003 PM 10651766 ER PT J AU Frankel, AE Kreitman, RJ Sausville, EA AF Frankel, AE Kreitman, RJ Sausville, EA TI Targeted toxins SO CLINICAL CANCER RESEARCH LA English DT Review ID RICIN-A-CHAIN; B-CELL LYMPHOMA; PHASE-I TRIAL; VASCULAR LEAK SYNDROME; PSEUDOMONAS EXOTOXIN; CONTINUOUS-INFUSION; ANTIBODY-RICIN; ANTI-B4-BLOCKED RICIN; ANTI-CD22 IMMUNOTOXIN; ANTILEUKEMIC ACTIVITY AB Targeted toxins, consisting of tumor-selective ligands coupled to polypeptide toxins, represent a nem class of cancer therapeutics that kills malignant cells by inactivating cytosolic protein synthesis and inducing apoptosis, A number of these molecules have been produced under good manufacturing practice conditions and given systemically to patients with a variety of neoplasms, The promising results to date and the remaining pharmacological hurdles are discussed. C1 Wake Forest Univ, Bowman Gray Sch Med, Winston Salem, NC 27157 USA. NCI, Med Branch, Clin Trials Unit, Mol Biol Lab,NIH, Bethesda, MD 20892 USA. NCI, Med Branch, Clin Trials Unit, Dev Therapeut Program,NIH, Bethesda, MD 20892 USA. RP Frankel, AE (reprint author), Wake Forest Univ, Bowman Gray Sch Med, Hanes 4046,Med Ctr Dr, Winston Salem, NC 27157 USA. NR 58 TC 130 Z9 144 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD FEB PY 2000 VL 6 IS 2 BP 326 EP 334 PG 9 WC Oncology SC Oncology GA 287JK UT WOS:000085502600003 PM 10690507 ER PT J AU Ornstein, DK Englert, C Gillespie, JW Paweletz, CP Linehan, WM Emmert-Buck, MR Petricoin, EF AF Ornstein, DK Englert, C Gillespie, JW Paweletz, CP Linehan, WM Emmert-Buck, MR Petricoin, EF TI Characterization of intracellular prostate-specific antigen from laser capture microdissected benign and malignant prostatic epithelium SO CLINICAL CANCER RESEARCH LA English DT Article ID MULTICENTER CLINICAL-TRIAL; CANCER; ALPHA-1-ANTICHYMOTRYPSIN; SERUM; COMPLEX; TISSUE; CELLS AB The proportion of unbound serum prostate-specific antigen (PSA; percent-free PSA) is reported to be lower in men with prostate cancer compared to men with benign prostates (U. H. Stenman et al,, Cancer Res., 51: 222-226, 1991; H, Lilja et at, Clin, Chem,, 37: 1618-1625, 1991; D. L. Woodrum ed at, J. Urol., 159: 5-12, 1998; W. J. Catalona et al., J. Am. Med. Assoc., 279: 1542-1547, 1998), The majority of immunoreactive PSA in serum is complexed to alpha-1-antichymotrypsin (ACT), Two major mechanistic questions have previously been unknown: (a) Does PSA in human prostate cancer cells in tissue exist in a free or bound form? and (b) Is PSA produced by malignant cells in the free form because it has lost the ability to form a complex with ACT? Laser capture microdissection (LCM) enables the acquisition of pure populations of defined cell types from tissue (M. R. Emmert-Buck et al., Science, 274: 998-1001, 1996; R. F. Bonner et al,, Science, 278: 1481-1483, 1997), This technology provides a unique opportunity to study intracellular protein composition and structure from human cells, In this study, we used LCM to assess the bound versus free form of intracellular PSA in both benign and malignant epithelium procured from prostate tissue, One-dimensional and two-dimensional PAGE were performed on cellular lysates from LCM-procured benign and malignant prostate epithelium from frozen tissue specimens. Western blotting analysis of one-dimensional PAGE gels revealed a strong band at nl, 30,000 (expected molecular weight of unbound PSA) in ail cases demonstrating that the vast majority of intracellular tumor and normal PSA exists within cells in the "free" form. Binding studies showed that PSA recovered from LCM-procured cells retained the full ability to bind ACT, and two-dimensional PAGE Western analysis demonstrated that the PSA/ACT complex was stable under strong reducing conditions. We conclude that intracellular PSA exists in the "free" form and that binding to ACT occurs exclusively outside of the cell. C1 NCI, Urol Oncol Branch, NIH, Bethesda, MD 20892 USA. NCI, Canc Genome Anat Project, Off Director, NIH, Bethesda, MD 20892 USA. US FDA, Ctr Biol Evaluat & Res, Div Cytokine Biol, Bethesda, MD 20892 USA. Georgetown Univ, Dept Chem, Washington, DC 20057 USA. NCI, Pathol Lab, Pathogenet Unit, NIH, Bethesda, MD 20892 USA. RP Ornstein, DK (reprint author), NCI, Urol Oncol Branch, NIH, Bldg 10,Room 2B47, Bethesda, MD 20892 USA. NR 11 TC 65 Z9 73 U1 0 U2 5 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD FEB PY 2000 VL 6 IS 2 BP 353 EP 356 PG 4 WC Oncology SC Oncology GA 287JK UT WOS:000085502600006 PM 10690510 ER PT J AU Wilson, WH Sorbara, L Figg, WD Mont, EK Sausville, E Warren, KE Balis, FM Bauer, K Raffeld, M Senderowicz, AM Monks, A AF Wilson, WH Sorbara, L Figg, WD Mont, EK Sausville, E Warren, KE Balis, FM Bauer, K Raffeld, M Senderowicz, AM Monks, A TI Modulation of clinical drug resistance in a B cell lymphoma patient by the protein kinase inhibitor 7-hydroxystaurosporine: Presentation of a novel therapeutic paradigm SO CLINICAL CANCER RESEARCH LA English DT Article ID SELECTIVE INHIBITOR; EPOCH CHEMOTHERAPY; ANTITUMOR-ACTIVITY; CANCER-THERAPY; IN-VIVO; UCN-01; APOPTOSIS; CYTOTOXICITY; P53; ARREST AB Emerging evidence suggests that apoptosis is an important mechanism of tumor cell death from antineoplastic therapy. 7-hydroxystaurosporine (UCN-01) is a novel protein kinase inhibitor that increases chemotherapy-induced apoptosis in vitro and is in early phases of clinical development. In this report, we present a 68-year-old patient with chemotherapy-resistant lymphoma treated with UCN-01 and chemotherapy, He had a stage IV plasmacytoid lymphoma that failed to enter remission with high-dose EPOCH LI (etoposide, prednisone, vincristine, cyclophosphamide, doxorubicin) chemotherapy, Due to disease progression and transformation to large cell lymphoma in the liver and bone marrow, he received UCN-01, Four weeks later, he received "standard-dose" EPOCH because of progression, developed severe neutropenia for 9 days, and expired from Candida sepsis on day 23, At autopsy, there was no histological evidence of residual lymphoma, although PCR for immunoglobulin gene rearrangement analysis revealed a faint clonal band in two of six nodes but none in the liver. Significantly, no B cells were detected by immunohistochemistry in lymph nodes, and a polyclonal ladder pattern associated with the presence of normal B cells was not seen in the immunoglobulin gene rearrangement PCR assay. Profound peripheral lymphopenia (50 cells/mu I) was also observed. Pharmacokinetics showed UCN-01 salivary concentrations, a surrogate for free drug concentrations, to be within an effective range in vitro (45 mmol/L) as a modulator of DNA-damaging agent cytotoxicity. In vitro, UCN-01 is synergistic with multiple cytotoxic agents and increases fludarabine-induced apoptosis in a human breast cell line. These results suggest that UCN-01 sensitized the lymphoma to the cytotoxic effects of EPOCH, possibly by modulating the "threshold" for apoptosis, and may illustrate a new paradigm for reversal of drug resistance. C1 NCI, Med Branch, Bethesda, MD 20892 USA. NCI, Pathol Lab, Bethesda, MD 20892 USA. NCI, Pediat Oncol Branch, Bethesda, MD 20892 USA. NCI, Dev Therapeut Program, Bethesda, MD 20892 USA. RP Wilson, WH (reprint author), NCI, Med Branch, Bldg 10,Room 12N-226,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Figg Sr, William/M-2411-2016 NR 32 TC 33 Z9 33 U1 0 U2 2 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD FEB PY 2000 VL 6 IS 2 BP 415 EP 421 PG 7 WC Oncology SC Oncology GA 287JK UT WOS:000085502600014 PM 10690518 ER PT J AU Robbins, DH Marguiles, I Stetler-Stevenson, M Kreitman, RJ AF Robbins, DH Marguiles, I Stetler-Stevenson, M Kreitman, RJ TI Hairy cell leukemia, a B-cell neoplasm that is particularly sensitive to the cytotoxic effect of anti-Tac(Fv)-PE38 (LMB-2) SO CLINICAL CANCER RESEARCH LA English DT Article ID SINGLE-CHAIN IMMUNOTOXIN; PSEUDOMONAS-EXOTOXIN; INTERLEUKIN-2 RECEPTOR; DIPHTHERIA-TOXIN; ANTI-TAC; PHASE-I; RECOMBINANT IMMUNOTOXINS; MONOCLONAL-ANTIBODY; CONTINUOUS-INFUSION; FV IMMUNOTOXINS AB Anti-Tac(Fv)-PE38 (LMB-2) is a recombinant, single-chain immunotoxin composed of the variable domains of the anti-Tac (anti-CD25) monoclonal antibody fused to a truncated form of Pseudomonas exotoxin (PE), Until now, this agent has been reported to be very cytotoxic toward T-cell but not B-cell leukemic cells freshly obtained from patients and is being tested clinically in patients with CD25+ malignancies of both B- and T-cell origin. Hairy cell leukemia (HCL) is a B-cell malignancy in which the cells are usually CD25+ and their ex vivo sensitivity to LMB-2 was unknown. Malignant cells from the first HCL patient to be tested were very sensitive to the cytotoxic effect of LMB-2 in vitro (IC50 1.1 ng/ml), and this patient responded clinically to LMB-2 administered systemically, Therefore, we decided to assess the potential clinical utility of LMB-2 in other patients with HCL, We tested fresh leukemic cells from nine additional CD25+ HCL patients. LMB-2 was very cytotoxic ex vivo in all patients with IC(50)s as low as 0.5 ng/ml. Malignant cells freshly obtained from patients with chronic lymphocytic leukemia were also sensitive to LMB-2 but not as sensitive as cells from HCL patients, These results indicate that CD25+ HCL is a B-cell neoplasm that is particularly sensitive to LMB-2, and this agent may be useful in patients who have faded standard therapies. C1 NCI, Clin Pathol Lab, Div Canc Therapy, NIH, Bethesda, MD 20892 USA. RP Kreitman, RJ (reprint author), NCI, Mol Biol Lab, Div Canc Biol, NIH, 37-4B27,37 Convent Dr,MSC 4255, Bethesda, MD 20892 USA. NR 48 TC 35 Z9 39 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1078-0432 J9 CLIN CANCER RES JI Clin. Cancer Res. PD FEB PY 2000 VL 6 IS 2 BP 693 EP 700 PG 8 WC Oncology SC Oncology GA 287JK UT WOS:000085502600051 PM 10690555 ER PT J AU Amemiya, K Semino-Mora, C Granger, RP Dalakas, MC AF Amemiya, K Semino-Mora, C Granger, RP Dalakas, MC TI Downregulation of TGF-beta 1 mRNA and protein in the muscles of patients with inflammatory myopathies after treatment with high-dose intravenous immunoglobulin SO CLINICAL IMMUNOLOGY LA English DT Article DE inflammatory myopathies; TGF-beta 1; intravenous immunoglobulin ID GROWTH-FACTOR-BETA; GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE; TRANSFORMING GROWTH-FACTOR-BETA-1; DERMATOMYOSITIS; CYTOKINE; GLOBULIN; FIBROSIS; GENE; EXPRESSION; DISEASES AB We used reverse(1) transcription-polymerase chain reaction to study the level of TGF-beta 1 mRNA expression and immunocytochemistry to examine the immunoreactive TGF-beta 1 in muscle biopsy specimens from five patients with dermatomyositis (DM) and five patients with inclusion body myositis (IBM) obtained before and after 3 months treatment with intravenous immunoglobulin (Mg). At baseline, the mRNA expression of TGF-beta 1 was increased up to fivefold in the muscles of DM patients compared to that of IBM patients. After IVIg, TGF-beta 1 was downregulated and the TGF-PI mRNA decreased twofold in the muscles of patients with DM who had successfully responded to therapy, but remained unchanged in the muscles of patients with IBM who did not respond. The downregulation of TGF-beta 1 in DM was associated with improvement of the muscle cytoarchitecture and reduction of the endomysial inflammation and connective tissue, suggesting that in DM the excess of TGF-beta 1 may be involved in the pathogenesis of chronic inflammation, fibrosis, and accumulation of extracellullar matrix proteins. (C) 2000 Academic Press. C1 NINDS, Neuromuscular Dis Sect, NIH, Bethesda, MD 20892 USA. RP Dalakas, MC (reprint author), NINDS, Neuromuscular Dis Sect, NIH, Bldg 10,Room 4N248,10 Ctr Dr,MSC 1382, Bethesda, MD 20892 USA. NR 20 TC 78 Z9 80 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1521-6616 J9 CLIN IMMUNOL JI Clin. Immunol. PD FEB PY 2000 VL 94 IS 2 BP 99 EP 104 DI 10.1006/clim.1999.4823 PG 6 WC Immunology SC Immunology GA 278UN UT WOS:000085007500003 PM 10637094 ER PT J AU Falloon, J Piscitelli, S Vogel, S Sadler, B Mitsuya, H Kavlick, MF Yoshimura, K Rogers, M LaFon, S Manion, DJ Lane, HC Masur, H AF Falloon, J Piscitelli, S Vogel, S Sadler, B Mitsuya, H Kavlick, MF Yoshimura, K Rogers, M LaFon, S Manion, DJ Lane, HC Masur, H TI Combination therapy with amprenavir, abacavir, and efavirenz in human immunodeficiency virus (HIV)-infected patients failing a protease-inhibitor regimen: Pharmacokinetic drug interactions and antiviral activity SO CLINICAL INFECTIOUS DISEASES LA English DT Article ID TYPE-1 VARIANTS; HIV-INFECTION; RESISTANCE; SENSITIVITY AB Patients with plasma viral RNA >50,000 copies/mL, despite a protease-inhibitor regimen, received abacavir, amprenavir, and efavirenz to assess efavirenz-amprenavir drug interactions and to evaluate safety and antiviral response. Patients first received amprenavir with abacavir and other nucleoside analogs. Amprenavir levels were measured before and after adding efavirenz. Patients then received a second protease inhibitor, There was evidence of genotypic and phenotypic resistance at study entry. No patient had study drugs discontinued because of toxicity. Efavirenz decreased the steady-state area under the curve, maximum plasma concentration, and minimum plasma concentration of amprenavir by 24%, 33%, and 43%, respectively. Three of 10 patients had >1.5 log(10) viral response to abacavir and amprenavir. All 8 patients who added efavirenz had >0.5 log(10) decline in viral load, and this response lasted >24 weeks for 3 of the patients. A combination regimen that included abacavir, amprenavir, and efavirenz was well tolerated and had sustained activity in some patients, Concomitant efavirenz therapy decreases amprenavir concentrations. C1 NIAID, NIH, Bethesda, MD 20892 USA. NCI, NIH, Bethesda, MD 20892 USA. NIH, Warren G Magnuson Clin Ctr, Bethesda, MD 20892 USA. Dupont Merck Pharmaceut Co, Wilmington, DE 19880 USA. RP Falloon, J (reprint author), NIAID, NIH, Bldg 10,Room 11C103,10 Ctr Dr MSC 1880, Bethesda, MD 20892 USA. NR 19 TC 60 Z9 60 U1 0 U2 3 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 1058-4838 J9 CLIN INFECT DIS JI Clin. Infect. Dis. PD FEB PY 2000 VL 30 IS 2 BP 313 EP 318 DI 10.1086/313667 PG 6 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 293JX UT WOS:000085849400016 PM 10671334 ER PT J AU Mima, T Steger, J Schulman, AE Gerloff, C Hallett, M AF Mima, T Steger, J Schulman, AE Gerloff, C Hallett, M TI Electroencephalographic measurement of motor cortex control of muscle activity in humans SO CLINICAL NEUROPHYSIOLOGY LA English DT Article DE EEG; coherence; cortical-muscular coupling; phase spectra; motor control; oscillatory activity ID EVENT-RELATED DESYNCHRONIZATION; FIELD POTENTIAL OSCILLATIONS; INFERIOR OLIVARY NEURONS; ISOMETRIC CONTRACTION; SENSORIMOTOR CORTEX; CORTICAL POTENTIALS; CORTICOSPINAL PROJECTIONS; COHERENT OSCILLATIONS; NEURAL ASSEMBLIES; HAND MOVEMENTS AB Objective: To detect and measure correlation between cortical and muscle activities, coherence analysis was used. Methods: The electroencephalogram (EEG) and electromyogram (EMG) were recorded in 9 normal volunteers during tonic contraction of upper and lower limb muscles on the right side. Coherence between EEG and EMG was computed to analyze their linear association. Results: EEG over the contralateral sensorimotor area was coherent with EMG, with peak coherence at 11-36 Hz (mean, 22 Hz). For the abductor pollicis brevis (APB) muscle, peak coherence, as determined by functional brain mapping with Focal transcranial magnetic stimulation (TMS), was over or slightly posterior to the hand area on the primary motor cortex determined by focal transcranial magnetic stimulation (TMS). Peak coherence over the scalp was somatotopically organized. The temporal relation between EEC and EMG was analyzed with a new model for interpreting the phase shift ('constant phase shift plus constant time lag' model). For the APE muscle, the phase relation between cortical and muscular oscillations differed in the frequency ranges of 3-13 Hz and 14-50 Hz, respectively, suggesting that different coupling mechanisms operate in different bands. Only the phase shift between cortical and motoneuronal firing at 14-50 Hz was reliably estimated by a linear model. At 14-50 Hz, motoneuronal firing was led by surface-negative cortical activity with a constant time lag that depended on the cortical-muscular distance. For the APE muscle, the time lag was slightly shorter than the cortical-muscular conduction time determined by TMS. Vibratory stimulation (100 Hz) of a muscle tendon during tonic contraction had no significant effect on cortical-muscular coherence. indicating that cortical oscillation reflected motor rather than sensory activity. Conclusions: The present findings suggest temporal coding of the oscillatory motor control system (3-13 Hz vs. 14-50 Hz), and confirm the functional importance of cortical beta and gamma rhythms in the motor efferent command. Cortical-muscular synchronization is most likely mediated by the direct corticospinal pathway within the frequency range of 14-50 Hz. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved. C1 NINDS, Human Motor Control Sect, Med Neurol Branch, NIH, Bethesda, MD 20892 USA. NINDS, Biometry & Field Studies Branch, NIH, Bethesda, MD 20892 USA. RP Hallett, M (reprint author), NINDS, Human Motor Control Sect, Med Neurol Branch, NIH, 10 Ctr Dr,MSC-1428, Bethesda, MD 20892 USA. OI Mima, Tatsuya/0000-0001-7787-4855 NR 70 TC 139 Z9 139 U1 4 U2 14 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 1388-2457 J9 CLIN NEUROPHYSIOL JI Clin. Neurophysiol. PD FEB PY 2000 VL 111 IS 2 BP 326 EP 337 DI 10.1016/S1388-2457(99)00229-1 PG 12 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 278UU UT WOS:000085008000017 PM 10680569 ER PT J AU Muellbacher, W Facchini, S Boroojerdi, B Hallett, M AF Muellbacher, W Facchini, S Boroojerdi, B Hallett, M TI Changes in motor cortex excitability during ipsilateral hand muscle activation in humans SO CLINICAL NEUROPHYSIOLOGY LA English DT Article DE transcranial magnetic stimulation; ipsilateral motor cortex; intracortical excitability; facilitation; motor evoked potentials ID TRANSCRANIAL MAGNETIC STIMULATION; VOLUNTARY CONTRACTION; CORTICOCORTICAL INHIBITION; HUMAN-BRAIN; FACILITATION; RESPONSES; POTENTIALS; CONDUCTION AB Objectives: To test whether unilateral hand muscle activation involves changes in ipsilateral primary motor cortex (M1) excitability. Methods: Single- and paired-pulse transcranial magnetic stimulation (TMS) of the right hemisphere was used to evoke motor evoked potentials (MEPs) from the resting left abductor pollicis brevis (APB) in 9 normal volunteers. We monitored changes in motor threshold (MT), MEP recruitment, intracortical inhibition (ICI) and intracortical facilitation (ICF) while the ipsilateral right APE was either at rest or voluntarily activated. Spinal motoneuron excitability was assessed using F-wave recording procedures. Results: Voluntary muscle activation of the ipsilateral APE significantly facilitated the MEPs and F-waves recorded from the contralateral APE. Facilitation was observed with muscle activation >50% of the maximum voluntary force and with stimulus intensities >20% above the individual resting motor threshold. Intracortical inhibition significantly decreased in the ipsilateral M1, while there was no significant change in intracortical facilitation during this maneuver. Conclusions: Unilateral hand muscle activation changes the excitability of homotopic hand muscle representations in both the ipsilateral M1 and the contralateral spinal cord. While the large proportion of MEP facilitation most likely occurred at a spinal level, involvement of the ipsilateral hemisphere may have contributed to the enlargement of magnetic responses. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved. C1 NINDS, Human Motor Control Sect, Med Neurol Branch, NIH, Bethesda, MD 20892 USA. RP Hallett, M (reprint author), NINDS, Human Motor Control Sect, Med Neurol Branch, NIH, Bldg 10,Room 5N226,10 Ctr Dr MSC-1428, Bethesda, MD 20892 USA. NR 22 TC 161 Z9 166 U1 0 U2 9 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 1388-2457 J9 CLIN NEUROPHYSIOL JI Clin. Neurophysiol. PD FEB PY 2000 VL 111 IS 2 BP 344 EP 349 DI 10.1016/S1388-2457(99)00243-6 PG 6 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 278UU UT WOS:000085008000019 PM 10680571 ER PT J AU Dionne, RA Jaber, L Gilron, I Brahim, JS Rowan, JS AF Dionne, RA Jaber, L Gilron, I Brahim, JS Rowan, JS TI Effect of the NK2 antagonist SR 48968C on acute pain and inflammation. SO CLINICAL PHARMACOLOGY & THERAPEUTICS LA English DT Meeting Abstract C1 Natl Inst Dent & Craniofacial Res, NIH, Bethesda, MD USA. NR 0 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 USA SN 0009-9236 J9 CLIN PHARMACOL THER JI Clin. Pharmacol. Ther. PD FEB PY 2000 VL 67 IS 2 MA PI13 BP 92 EP 92 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 288CA UT WOS:000085543200014 ER PT J AU Dionne, RA Brahim, JS Rowan, JS Guivarch, PH AF Dionne, RA Brahim, JS Rowan, JS Guivarch, PH TI Analgesic effect of microencapsulated flurbiprofen in the oral surgery model of acute pain. SO CLINICAL PHARMACOLOGY & THERAPEUTICS LA English DT Meeting Abstract C1 RTP Pharma, Quebec City, PQ, Canada. Natl Inst Dent & Craniofacial Res, NIH, Bethesda, MD USA. NR 0 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 USA SN 0009-9236 J9 CLIN PHARMACOL THER JI Clin. Pharmacol. Ther. PD FEB PY 2000 VL 67 IS 2 MA PI12 BP 92 EP 92 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 288CA UT WOS:000085543200013 ER PT J AU Gilron, I Max, MB Lee, G Booher, SL Dionne, RA AF Gilron, I Max, MB Lee, G Booher, SL Dionne, RA TI Safety and analgesic efficacy of the AMPA/kainate antagonist LY293558 following oral surgery. SO CLINICAL PHARMACOLOGY & THERAPEUTICS LA English DT Meeting Abstract C1 Natl Inst Dent & Craniofacial Res, Pain & Neurosensory Mech Branch, NIH, Bethesda, MD USA. NR 0 TC 1 Z9 1 U1 0 U2 0 PU MOSBY-YEAR BOOK INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0009-9236 J9 CLIN PHARMACOL THER JI Clin. Pharmacol. Ther. PD FEB PY 2000 VL 67 IS 2 MA PI16 BP 93 EP 93 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 288CA UT WOS:000085543200017 ER PT J AU Kearns, GL Stalker, D Blumer, JL James, LP Jungbluth, G Reed, MD Jacobs, RF Abdel-Rahman, S AF Kearns, GL Stalker, D Blumer, JL James, LP Jungbluth, G Reed, MD Jacobs, RF Abdel-Rahman, S TI Linezolid disposition in infants and children. SO CLINICAL PHARMACOLOGY & THERAPEUTICS LA English DT Meeting Abstract C1 NICHD, Bethesda, MD USA. Univ Arkansas Med Sci, Little Rock, AR 72205 USA. Case Western Reserve Univ, Cleveland, OH 44106 USA. Pharmacia & Upjohn Inc, Kalamazoo, MI 49001 USA. Childrens Mercy Hosp, Kansas City, MO 64108 USA. NR 0 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 USA SN 0009-9236 J9 CLIN PHARMACOL THER JI Clin. Pharmacol. Ther. PD FEB PY 2000 VL 67 IS 2 MA PI65 BP 105 EP 105 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 288CA UT WOS:000085543200065 ER PT J AU Aszalos, A Ibrahim, S Knapton, A Licht, T AF Aszalos, A Ibrahim, S Knapton, A Licht, T TI Reversal of P-glycoprotein-mediated multidrug resistance in vitro by antihistamines and diuretics. SO CLINICAL PHARMACOLOGY & THERAPEUTICS LA English DT Meeting Abstract C1 NCI, NIH, Bethesda, MD 20892 USA. US FDA, Ctr Drug Evaluat & Res, Laurenceville, MD USA. NR 0 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 USA SN 0009-9236 J9 CLIN PHARMACOL THER JI Clin. Pharmacol. Ther. PD FEB PY 2000 VL 67 IS 2 MA PI70 BP 106 EP 106 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 288CA UT WOS:000085543200070 ER PT J AU Economou, AP Yang, HYT Iadarola, MI Dionne, RA AF Economou, AP Yang, HYT Iadarola, MI Dionne, RA TI Expression of COX-2 in tissue biopsies following oral surgery. SO CLINICAL PHARMACOLOGY & THERAPEUTICS LA English DT Meeting Abstract C1 Natl Inst Dent & Craniofacial Res, NIH, Bethesda, MD USA. NR 0 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 USA SN 0009-9236 J9 CLIN PHARMACOL THER JI Clin. Pharmacol. Ther. PD FEB PY 2000 VL 67 IS 2 MA OIB2 BP 114 EP 114 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 288CA UT WOS:000085543200100 ER PT J AU Yu, Z Huse, LM Engler, MM Hammock, BD Zeldin, DC Kroetz, DL AF Yu, Z Huse, LM Engler, MM Hammock, BD Zeldin, DC Kroetz, DL TI Antihypertensive effect of soluble epoxide hydrolase (sEH) inhibition. SO CLINICAL PHARMACOLOGY & THERAPEUTICS LA English DT Meeting Abstract C1 NIEHS, Res Triangle Pk, NC 27709 USA. Univ Calif Davis, Dept Entomol, Davis, CA 95616 USA. Univ Calif San Francisco, Dept Biopharmaceut Sci, San Francisco, CA 94143 USA. Univ Calif San Francisco, Dept Physiol Nursing, San Francisco, CA 94143 USA. NR 0 TC 0 Z9 0 U1 0 U2 1 PU MOSBY-YEAR BOOK INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0009-9236 J9 CLIN PHARMACOL THER JI Clin. Pharmacol. Ther. PD FEB PY 2000 VL 67 IS 2 MA PII53 BP 128 EP 128 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 288CA UT WOS:000085543200155 ER PT J AU Ibrahim, S Knapton, A Licht, T Aszalos, A AF Ibrahim, S Knapton, A Licht, T Aszalos, A TI Modulation of the function of P-glycoprotein in NIH3T3 and human capillary endothelial cells by Ca2+ channel blockers. SO CLINICAL PHARMACOLOGY & THERAPEUTICS LA English DT Meeting Abstract C1 NCI, NIH, Bethesda, MD 20892 USA. US FDA, Ctr Drug Evaluat & Res, Rockville, MD 20857 USA. NR 0 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 USA SN 0009-9236 J9 CLIN PHARMACOL THER JI Clin. Pharmacol. Ther. PD FEB PY 2000 VL 67 IS 2 MA PII79 BP 134 EP 134 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 288CA UT WOS:000085543200181 ER PT J AU Kent, AA Brahim, JS Gilron, I Rowan, JS Dionne, RA AF Kent, AA Brahim, JS Gilron, I Rowan, JS Dionne, RA TI Effect of dexamethasone on PGE(2) and thromboxane B-2 levels following tissue injury. SO CLINICAL PHARMACOLOGY & THERAPEUTICS LA English DT Meeting Abstract C1 Natl Inst Dent & Craniofacial Res, NIH, Bethesda, MD USA. NR 0 TC 1 Z9 1 U1 0 U2 0 PU MOSBY-YEAR BOOK INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0009-9236 J9 CLIN PHARMACOL THER JI Clin. Pharmacol. Ther. PD FEB PY 2000 VL 67 IS 2 MA PIII5 BP 143 EP 143 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 288CA UT WOS:000085543200216 ER PT J AU Marshall, JD Rodarte, A Blumer, JL Khoo, K Akbari, B Kearns, GL AF Marshall, JD Rodarte, A Blumer, JL Khoo, K Akbari, B Kearns, GL CA PPRU Network TI Pediatric pharmacodynamics of midazolam oral syrup. SO CLINICAL PHARMACOLOGY & THERAPEUTICS LA English DT Meeting Abstract C1 NICHD, Bethesda, MD USA. Roche Labs Inc, Nutley, NJ USA. Case Western Reserve Univ, Cleveland, OH 44106 USA. Univ Calif San Diego, La Jolla, CA 92093 USA. Childrens Mercy Hosp, Kansas City, MO 64108 USA. NR 0 TC 0 Z9 0 U1 0 U2 1 PU MOSBY-YEAR BOOK INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0009-9236 J9 CLIN PHARMACOL THER JI Clin. Pharmacol. Ther. PD FEB PY 2000 VL 67 IS 2 MA PIII6 BP 143 EP 143 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 288CA UT WOS:000085543200217 ER PT J AU Thevanayagam, L Hayashi, S Watts, H Gambertoglio, J Aberg, J Shelton, M Lizak, P Aweeka, F AF Thevanayagam, L Hayashi, S Watts, H Gambertoglio, J Aberg, J Shelton, M Lizak, P Aweeka, F TI The impact of gender and contraceptive therapy on the pharmacokinetics (PK) of plasma (P) and intracellular (IC) zidovudine (Z). ACTG protocol 317. SO CLINICAL PHARMACOLOGY & THERAPEUTICS LA English DT Meeting Abstract C1 NICHD, NIH, Bethesda, MD 20892 USA. SUNY Buffalo, Dept Pharm Practice & Med, Buffalo, NY USA. Univ Calif San Francisco, San Francisco Gen Hosp, Dept Med, San Francisco, CA 94143 USA. Univ Calif San Francisco, San Francisco Gen Hosp, Dept Clin Pharm, San Francisco, CA 94143 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI NEW YORK PA 75 VARICK ST, 9TH FLR, NEW YORK, NY 10013-1917 USA SN 0009-9236 J9 CLIN PHARMACOL THER JI Clin. Pharmacol. Ther. PD FEB PY 2000 VL 67 IS 2 MA PIII76 BP 161 EP 161 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 288CA UT WOS:000085543200287 ER PT J AU Magee, MH Forrest, A Shelton, M Para, M Fischi, M Morse, G AF Magee, MH Forrest, A Shelton, M Para, M Fischi, M Morse, G TI An integrated PK/PD model for delavirdine (DLV). SO CLINICAL PHARMACOLOGY & THERAPEUTICS LA English DT Meeting Abstract C1 NIH, ACTG Protocol Team 260, Adult ACTG, Bethesda, MD 20892 USA. Univ Miami, Miami, FL 33152 USA. Ohio State Univ, Columbus, OH 43210 USA. SUNY Buffalo, Buffalo, NY USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI NEW YORK PA 75 VARICK ST, 9TH FLR, NEW YORK, NY 10013-1917 USA SN 0009-9236 J9 CLIN PHARMACOL THER JI Clin. Pharmacol. Ther. PD FEB PY 2000 VL 67 IS 2 MA PIII82 BP 162 EP 162 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 288CA UT WOS:000085543200293 ER PT J AU Leeder, JS Adcock, K Gaedigk, A Gotschall, R Wilson, JT Kearns, GL AF Leeder, JS Adcock, K Gaedigk, A Gotschall, R Wilson, JT Kearns, GL TI Acquisition of CYP2D6 and CYP3A activities in the first year of life. SO CLINICAL PHARMACOLOGY & THERAPEUTICS LA English DT Meeting Abstract C1 Childrens Mercy Hosp, Kansas City, MO 64108 USA. LSU, Med Ctr, NICHD, PPRUs, Shreveport, LA USA. NR 0 TC 5 Z9 5 U1 0 U2 0 PU MOSBY-YEAR BOOK INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0009-9236 J9 CLIN PHARMACOL THER JI Clin. Pharmacol. Ther. PD FEB PY 2000 VL 67 IS 2 MA OIIIB3 BP 169 EP 169 PG 1 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 288CA UT WOS:000085543200320 ER PT J AU Grady, CL McIntosh, AR Horwitz, B Rapoport, SI AF Grady, CL McIntosh, AR Horwitz, B Rapoport, SI TI Age-related changes in the neural correlates of degraded and nondegraded face processing SO COGNITIVE NEUROPSYCHOLOGY LA English DT Article ID POSITRON-EMISSION TOMOGRAPHY; CEREBRAL BLOOD-FLOW; HUMAN EXTRASTRIATE CORTEX; SUPERIOR TEMPORAL SULCUS; PRIMATE VISUAL-SYSTEM; PARTIAL LEAST-SQUARES; WORKING-MEMORY; HUMAN-BRAIN; FUNCTIONAL NEUROANATOMY; POSTERIOR PARIETAL AB In order to explore the neural correlates of age-related changes in visual perception of faces, positron emission tomographic scans were obtained on young and old adults while they were engaged in tasks of nondegraded and degraded face matching. Old adults were less accurate than were young adults across all face matching conditions, although the age difference was greatly reduced when degraded performance was adjusted for nondegraded performance. The interaction of age and degree of degradation on performance measures was not significant. Brain activity patterns during nondegraded face matching were similar in the two groups with some differences in parietal and prestriate cortices (greater activity in young adults) and in prefrontal cortex, thalamus, and hippocampus (greater activity in old adults). Increases in activity related to increasing degradation of the faces were seen mainly in prefrontal cortices in both age groups. Despite this similarity in the brain response to face degradation, there were striking differences between groups in the correlations between brain activity and degraded task performance. Different regions of extrastriate cortex were positively correlated with behavioural measures in the two groups (fusiform gyrus in. the young adults and posterior occipital regions in old adults). In addition two areas where older adults showed greater activity during nondegraded face matching, thalamus and hippocampus, also showed positive correlations with behaviour during the degraded tasks in this group, but not in the young group. Thus, although the elderly are not more vulnerable to the effects of increasing face degradation, the brain systems involved in carrying out these visual discriminations in young and old adults are not the same. These results are consistent with the idea of functional plasticity in face processing over the life span. C1 Univ Toronto, Baycrest Ctr Geriat Care, Rotman Res Inst, Toronto, ON M6A 2E1, Canada. NIA, Bethesda, MD 20892 USA. RP Grady, CL (reprint author), Univ Toronto, Baycrest Ctr Geriat Care, Rotman Res Inst, 3560 Bathurst St, Toronto, ON M6A 2E1, Canada. RI McIntosh, Anthony/G-4955-2011; OI McIntosh, Anthony/0000-0002-1784-5662 NR 73 TC 58 Z9 59 U1 0 U2 4 PU PSYCHOLOGY PRESS PI HOVE PA 27 CHURCH RD, HOVE BN3 2FA, EAST SUSSEX, ENGLAND SN 0264-3294 J9 COGNITIVE NEUROPSYCH JI Cogn. Neuropsychol. PD FEB-MAY PY 2000 VL 17 IS 1-3 BP 165 EP 186 PG 22 WC Psychology; Psychology, Experimental SC Psychology GA 305QC UT WOS:000086550900011 PM 20945178 ER PT J AU Gamel, JW Weller, EA Wesley, MN Feuer, EJ AF Gamel, JW Weller, EA Wesley, MN Feuer, EJ TI Parametric cure models of relative and cause-specific survival for grouped survival times SO COMPUTER METHODS AND PROGRAMS IN BIOMEDICINE LA English DT Article DE parametric cure model; relative survival; cause-specific survival; lognormal model; loglogistic model; Weibull model; gompertz ID TUMOR AB With parametric cure models, we can express survival parameters (e.g. cured fraction, location and scale parameters) as functions of covariates. These models can measure survival from a specific disease process, either by examining deaths due to the cause under study (cause-specific survival), or by comparing all deaths to those in a matched control population (relative survival). We present a binomial maximum likelihood algorithm to be used for actuarial data, where follow-up times are grouped into specific intervals. Our algorithm provides simultaneous maximum likelihood estimates for all the parameters of a cure model and can be used for cause-specific or relative survival analysis with a variety of survival distributions. Current software does not provide the flexibility of this unified approach. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved. C1 Vet Adm Hosp, Louisville, KY 40292 USA. Univ Louisville, Sch Med, Louisville, KY 40292 USA. Harvard Univ, Sch Publ Hlth, Boston, MA 02115 USA. Dana Farber Canc Inst, Boston, MA 02115 USA. Informat Management Serv, Silver Spring, MD 20904 USA. NCI, Div Canc Prevent & Control, Bethesda, MD 20892 USA. RP Gamel, JW (reprint author), Vet Adm Hosp, 301 E Muhammed Ali Blvd, Louisville, KY 40292 USA. NR 13 TC 9 Z9 10 U1 0 U2 1 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0169-2607 J9 COMPUT METH PROG BIO JI Comput. Meth. Programs Biomed. PD FEB PY 2000 VL 61 IS 2 BP 99 EP 110 DI 10.1016/S0169-2607(99)00022-X PG 12 WC Computer Science, Interdisciplinary Applications; Computer Science, Theory & Methods; Engineering, Biomedical; Medical Informatics SC Computer Science; Engineering; Medical Informatics GA 271XJ UT WOS:000084619900003 PM 10661395 ER PT J AU McCrae, RR AF McCrae, RR TI Personality and person perception across cultures SO CONTEMPORARY PSYCHOLOGY-APA REVIEW OF BOOKS LA English DT Book Review C1 NIA, Personal Stress & Coping Sect, Gerontol Res Ctr, NIH, Baltimore, MD 21224 USA. RP McCrae, RR (reprint author), NIA, Personal Stress & Coping Sect, Gerontol Res Ctr, NIH, POB 03,5600 Nathan Shock, Baltimore, MD 21224 USA. NR 11 TC 0 Z9 0 U1 0 U2 1 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 0010-7549 J9 CONTEMP PSYCHOL JI Comtemp. Psychol. PD FEB PY 2000 VL 45 IS 1 BP 38 EP 40 PG 3 WC Psychology, Multidisciplinary SC Psychology GA 289EW UT WOS:000085608400016 ER PT J AU Dixon, DO Lagakos, SW AF Dixon, DO Lagakos, SW TI Should data and safety monitoring boards share confidential interim data? SO CONTROLLED CLINICAL TRIALS LA English DT Article DE clinical trial; data sharing; DSMB procedures ID CLINICAL-TRIALS; ISSUES AB Consider the following situation: Two clinical trials are underway, closely related in terms of the interventions being compared and the target populations. In preparing for a planned interim analysis, the statistician for trial 1 finds that the results support a recommendation to stop the trial early. Should the statistician ask the investigators for trial 2 to make interim results of their trial available to the data and safety monitoring board (DSMB) for trial 1? If so, in what form? Would the answers change if the trial 1 results showed a strong but not convincing trend? What is the obligation of the trial 2 investigators to respond to such a request? What role do the two DSMBs have, either in initiating a request or in agreeing to respond to it? In this article, we examine this situation in some detail, having faced it occasionally in our own experience with clinical trials and DSMBs. The chief argument in favor of sharing data is that data from trial 2 are obviously relevant to the question being addressed by trial 1 and therefore ought to be available to those who must interpret the results from that trial. On the other hand, there are several reasons for not sharing interim data. For example, sharing is incompatible with the independence of the trials; the time for synthesizing evidence from both trials is after the two teams of investigators have presented the full analysis and interpretation of their separate trials. For this and other conceptual and practical reasons we conclude that it is better, in most cases, for DSMBs to consider only information that has already been made public in some form. Control Clin Trials 2000;21:1-6 (C) Elsevier Science Inc. 2000. C1 NIAID, Biostat Res Branch, Div AIDS, NIH,DAIDS, Bethesda, MD 20892 USA. Harvard Univ, Sch Publ Hlth, Dept Biostat, Boston, MA 02115 USA. RP Dixon, DO (reprint author), NIAID, Biostat Res Branch, Div AIDS, NIH,DAIDS, Rockledge 4115,6700-B Rockledge Dr MSC 7620, Bethesda, MD 20892 USA. FU NIAID NIH HHS [AI24643] NR 4 TC 13 Z9 13 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0197-2456 J9 CONTROL CLIN TRIALS JI Controlled Clin. Trials PD FEB PY 2000 VL 21 IS 1 BP 1 EP 6 DI 10.1016/S0197-2456(99)00042-2 PG 6 WC Medicine, Research & Experimental; Pharmacology & Pharmacy SC Research & Experimental Medicine; Pharmacology & Pharmacy GA 277AX UT WOS:000084911500001 PM 10659999 ER PT J AU Mattson, AJ Levin, HS Grafman, J AF Mattson, AJ Levin, HS Grafman, J TI A case of prosopagnosia following moderate closed head injury with left hemisphere focal lesion SO CORTEX LA English DT Article ID FACE RECOGNITION; OBJECT; PATHWAYS; DISSOCIATION; AGNOSIA; PET AB This study describes a left handed woman with prosopagnosia following traumatic brain injury with a focal lesion confined to the left-posterior hemisphere. Few cases of prosopagnosia following unilateral left hemisphere lesions have previously been reported in the literature. Corrected visual acuity was 20/70 (binocular), color vision was intact on screening, and shape detection was borderline. Impairments in higher order visual perception were evident to varying degrees on nonfacial tasks. Matching of unfamiliar faces was very slow but accurate. A marked impairment in the ability to recognize familiar faces and learn new face-name associations was evident on experimental tasks relative to the performance of healthy control subjects. In contrast, identification of characteristics of faces (gender, age) and identification and matching of facial expressions were relatively preserved. We discuss the cognitive processing stages that appear to be disrupted using Bruce and Young's (1986) model of facial recognition and perception as a framework. C1 Western State Hosp, Neuropsychol Serv, Steilacoom, WA USA. Baylor Coll Med, Dept Phys Med & Rehabil, Houston, TX 77030 USA. Baylor Coll Med, Dept Psychiat & Behav Sci, Houston, TX 77030 USA. Baylor Coll Med, Dept Neurosurg, Houston, TX 77030 USA. NIH, Cognit Neurosci Sect, Med Neurol Branch, Bethesda, MD 20892 USA. RP Levin, HS (reprint author), PM&R Res Off, 1333 Moursund Ave,Room A205, Houston, TX 77003 USA. OI Grafman, Jordan H./0000-0001-8645-4457 FU NINDS NIH HHS [NS-21889] NR 36 TC 48 Z9 49 U1 1 U2 2 PU MASSON DIVISIONE PERIODICI PI MILAN PA VIA FRATELLI BRESSAN 2, 20126 MILAN, ITALY SN 0010-9452 J9 CORTEX JI Cortex PD FEB PY 2000 VL 36 IS 1 BP 125 EP 137 DI 10.1016/S0010-9452(08)70841-4 PG 13 WC Behavioral Sciences; Neurosciences SC Behavioral Sciences; Neurosciences & Neurology GA 291MN UT WOS:000085740800010 PM 10728902 ER PT J AU DePietro, MR Eichacker, PQ AF DePietro, MR Eichacker, PQ TI Lidocaine for acute lung injury: Questions still to answer SO CRITICAL CARE MEDICINE LA English DT Editorial Material DE immune response, neutrophil; lung, edema; acute respiratory distress syndrome; pancreatitis; anesthetics, lidocaine ID ACUTE MYOCARDIAL-INFARCTION; ENDOTOXIN-SHOCK; SURVIVAL; INDOMETHACIN; SEPSIS; RATS C1 NIH, Bethesda, MD 20892 USA. RP DePietro, MR (reprint author), NIH, Bethesda, MD 20892 USA. NR 25 TC 3 Z9 6 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0090-3493 J9 CRIT CARE MED JI Crit. Care Med. PD FEB PY 2000 VL 28 IS 2 BP 589 EP 591 DI 10.1097/00003246-200002000-00060 PG 3 WC Critical Care Medicine SC General & Internal Medicine GA 288CB UT WOS:000085543300060 PM 10708215 ER PT J AU Ambudkar, IS AF Ambudkar, IS TI Regulation of calcium in salivary gland secretion SO CRITICAL REVIEWS IN ORAL BIOLOGY & MEDICINE LA English DT Review DE calcium homeostasis; calcium influx; ion channels; fluid secretion salivary gland; physiology ID PAROTID ACINAR-CELLS; INOSITOL 1,4,5-TRISPHOSPHATE RECEPTOR; BASOLATERAL PLASMA-MEMBRANE; DIVALENT-CATION ENTRY; CAPACITATIVE CA2+ ENTRY; DEPENDENT CA-2+ TRANSPORT; SUBLINGUAL MUCOUS ACINI; RAT MAST-CELLS; PANCREATIC ACINAR; ENDOPLASMIC-RETICULUM AB Neurotransmitter-regulation of fluid secretion in the salivary glands is achieved by a coordinated sequence of intracellular signaling events, including the activation of membrane receptors, generation of the intracellular second messenger, inositol 1,4,5, trisphosphate, internal Ca2+ release, and Ca2+ influx. The resulting increase in cytosolic [Ca2+] ([Ca2+](i)) regulates a number of ion transporters, e.g., Ca2+-activated K+ channel, Na+/K+/2Cl(-) co-transporter in the basolateral membrane, and the Ca2+-activated Cl- channel in the luminal membrane, which are intricately involved in fluid secretion. Thus, regulation of [Ca2+](i) is central to the regulation of salivary acinar cell function and is achieved by the concerted activities of several ion channels and Ca2+-pumps localized in various cellular membranes. Ca2+ pumps, present in the endoplasmic reticulum and the plasma membrane, serve to remove Ca2+ from the cytosol. Ca2+ channels present in the endoplasmic reticulum and the plasma membrane facilitate rapid influx of Ca2+ into the cytosol from the internal Ca2+ stores and from the external medium, respectively. It is well-established that prolonged fluid secretion is regulated via a sustained elevation in [Ca2+](i) that is primarily achieved by the influx of Ca2+ into the cell from the external medium. This Ca2+ influx occurs via a putative plasma-membrane-store-operated Ca2+ channel which has not yet been identified in any non-excitable cell type. Understanding the molecular nature of this Ca2+ influx mechanism is critical to our understanding of Ca2+ signaling in salivary gland cells. This review focuses on the various active and passive Ca2+ transport mechanisms in salivary gland cells-their localization, regulation, and role in neurotransmitter-regulation of fluid secretion. In addition to a historical perspective of Ca2+ signaling, recent findings and challenging problems facing this field are highlighted. C1 Natl Inst Dent & Craniofacial Res, Gene Therapy & Therapeut Branch, Secretory Physiol Sect, NIH, Bethesda, MD 20892 USA. RP Ambudkar, IS (reprint author), Natl Inst Dent & Craniofacial Res, Gene Therapy & Therapeut Branch, Secretory Physiol Sect, NIH, Bldg 10,Room 1N-113, Bethesda, MD 20892 USA. NR 175 TC 68 Z9 69 U1 0 U2 2 PU INT AMER ASSOC DENTAL RESEARCH PI ALEXANDRIA PA 1619 DUKE ST, ALEXANDRIA, VA 22314-3406 USA SN 1045-4411 J9 CRIT REV ORAL BIOL M JI Crit. Rev. Oral Biol. Med. PD FEB PY 2000 VL 11 IS 1 BP 4 EP 25 PG 22 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA 313PJ UT WOS:000087007800001 PM 10682899 ER PT J AU Frederikse, PH Zigler, JS Farnsworth, PN Carper, DA AF Frederikse, PH Zigler, JS Farnsworth, PN Carper, DA TI Prion protein expression in mammalian lenses SO CURRENT EYE RESEARCH LA English DT Article DE prion protein; lens; cataract; oxidative stress; Alzheimer pathophysiology ID INCLUSION-BODY MYOSITIS; AMYLOID PRECURSOR PROTEIN; CREUTZFELDT-JAKOB-DISEASE; VULNERABLE BRAIN-REGIONS; OXIDATIVE STRESS; NEUROMUSCULAR-JUNCTIONS; THIAMINE-DEFICIENCY; ALZHEIMERS-DISEASE; EPITHELIAL-CELLS; GENE-EXPRESSION AB Purpose. Aging and oxidative stress resulting from overexpression of Alzheimer precursor protein (beta APP) have been studied as important factors contributing to the major age-related (sporadic), and minor (hereditary) forms of Alzheimer's disease (AD), and muscle inclusion body myositis, (IBM). AD and prion proteins accumulate in plaques linked with AD and scrapie diseases, and in rimmed vacuoles of IBM. Soluble beta-amyloid (Ab) fibrillar forms are now thought to play a critical role in and outside of cells by producing oxidative stress. In lens, beta APP and Ab increase in cultured lenses exposed to oxidative stress, and in areas of lens fiber cell degeneration in thiamine (vitamin B1) deprived mice, a classic model of systemic oxidative stress. The purpose of the present study is to extend our studies of amyloid disease-related protein expression in mammalian lenses. Methods. Western blot, immunohistochemical detection, and RT-PCR methods were used to identify and quantitate prion protein expression in human, monkey, and guinea pig lenses. Results. We demonstrate for the first time that prion protein gene expression increases with oxidative stress in cultured human lens epithelial cells. In addition, we detected greater prion protein gene expression in fiber cells than epithelial cells in vivo. This is consistent with increases in prion protein expression demonstrated in myoblasts and neuronal cells induced to differentiate. Our initial investigations of prion protein in human lens cataracts identified increased prion protein immunoreactivity in regions of lens fiber cell degeneration. Conclusions. The present data indicate that prion protein expression increases during lens development, and is substantially increased in cultured human lens epithelial cells exposed to oxidative stress. We also provide evidence that prion protein immunoreactivity can be increased in regions of fiber cell disorganization. These data suggest a potential role for prion protein as a marker for some types of lens pathology, and in the mechanism of oxidative stress-related lens degeneration. C1 Univ Med & Dent New Jersey, New Jersey Med Sch, Dept Pharmacol & Physiol, Newark, NJ 07103 USA. Univ Med & Dent New Jersey, New Jersey Med Sch, Dept Ophthalmol, Newark, NJ 07103 USA. NEI, NIH, Lab Mech Ocular Dis, Bethesda, MD USA. RP Frederikse, PH (reprint author), Univ Med & Dent New Jersey, New Jersey Med Sch, Dept Pharmacol & Physiol, MSB H-645, Newark, NJ 07103 USA. NR 37 TC 24 Z9 25 U1 0 U2 1 PU SWETS ZEITLINGER PUBLISHERS PI LISSE PA P O BOX 825, 2160 SZ LISSE, NETHERLANDS SN 0271-3683 J9 CURR EYE RES JI Curr. Eye Res. PD FEB PY 2000 VL 20 IS 2 BP 137 EP 143 DI 10.1076/0271-3683(200002)20:2;1-D;FT137 PG 7 WC Ophthalmology SC Ophthalmology GA 287BM UT WOS:000085484900011 PM 10617916 ER PT J AU Brantner, CA Buchholz, LA Remsen, CC Collins, MLP AF Brantner, CA Buchholz, LA Remsen, CC Collins, MLP TI Isolation of intracytoplasmic membrane from the methanotrophic bacterium Methylomicrobium album BG8 SO CURRENT MICROBIOLOGY LA English DT Article ID METHYLOCOCCUS-CAPSULATUS BATH; PARTICULATE METHANE MONOOXYGENASE; RHODOSPIRILLUM-RUBRUM; OUTER-MEMBRANE; COPPER AB Methane-oxidizing bacteria, including Methylomicrobium album BG8, form an intracytoplasmic membrane in addition to the cytoplasmic and outer membranes of the cell envelope. Techniques to isolate the intracytoplasmic membrane of M. album BG8 were developed. An intracytoplasmic membrane fraction was separated from a cell envelope fraction on the basis of sedimentation velocity in sucrose density gradients. Proteins associated with the particulate methane monooxygenase were found in both membrane fractions. C1 Univ Wisconsin, Dept Biol Sci, Milwaukee, WI 53201 USA. Univ Wisconsin, Great Lakes Water Inst, Milwaukee, WI 53201 USA. RP Collins, MLP (reprint author), NINDS, NIH, 36 Convent Dr, Bethesda, MD 20892 USA. OI Brantner, Christine/0000-0001-8172-901X FU NIGMS NIH HHS [R15GM51006] NR 17 TC 4 Z9 4 U1 0 U2 5 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0343-8651 J9 CURR MICROBIOL JI Curr. Microbiol. PD FEB PY 2000 VL 40 IS 2 BP 132 EP 134 DI 10.1007/s002849910026 PG 3 WC Microbiology SC Microbiology GA 269XK UT WOS:000084502600012 PM 10594229 ER PT J AU Wu, XF Jung, G Hammer, JA AF Wu, XF Jung, G Hammer, JA TI Functions of unconventional myosins SO CURRENT OPINION IN CELL BIOLOGY LA English DT Review ID GTPASE-ACTIVATING PROTEIN; SACCHAROMYCES-CEREVISIAE; ORGANELLE TRANSPORT; LIGHT-CHAIN; IN-VIVO; DROSOPHILA EMBRYOS; ACTIN CYTOSKELETON; DISTINCT FUNCTIONS; POLARIZED GROWTH; HAIR-CELLS AB To date, fourteen classes of;unconventional myosins have been identified. Recent reports have implicated a number of these myosins in organelle transport, and in the formation, maintenance and/or dynamics of actin-rich structures involved in a variety of cellular processes including endocytosis, cell migration, and sensory transduction. Characterizations of organelle dynamics in pigment cells and neurons have further defined the contributions made by unconventional myosins and microtubule motors to the transport and distribution of organelles. Several studies have provided evidence of complexes through which cooperative organelle transport may be coordinated. Finally, the myosin superfamily has been shown to contain at least one processive motor and one backwards motor. C1 NIH, Cell Biol Lab, Sect Mol Cell Biol, Bethesda, MD 20892 USA. RP Wu, XF (reprint author), NIH, Cell Biol Lab, Sect Mol Cell Biol, Bldg 3,Room B1-22,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 106 TC 93 Z9 93 U1 2 U2 13 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0955-0674 J9 CURR OPIN CELL BIOL JI Curr. Opin. Cell Biol. PD FEB PY 2000 VL 12 IS 1 BP 42 EP 51 DI 10.1016/S0955-0674(99)00055-1 PG 10 WC Cell Biology SC Cell Biology GA 286XY UT WOS:000085475200005 PM 10679359 ER PT J AU Abrams, SI Schlom, J AF Abrams, SI Schlom, J TI Rational antigen modification as a strategy to upregulate or downregulate antigen recognition SO CURRENT OPINION IN IMMUNOLOGY LA English DT Review ID ALTERED PEPTIDE LIGAND; MHC CLASS-I; HUMAN CARCINOEMBRYONIC ANTIGEN; T-CELL EPITOPES; COMPLEX STABILITY; PARTIAL AGONISTS; CANCER VACCINES; CTL EPITOPE; IMMUNOGENICITY; IDENTIFICATION AB Recent and rapid advances in our understanding of the cellular and molecular mechanisms of antigen recognition by CD8(+) and CD4(+) T lymphocytes have led to the birth of possibilities for site-directed, rational modification of cognate antigenic determinants. This immunologic concept has vast biomedical implications for regulation of host immunity against the pathogenesis of diverse disease processes. The upregulation of antigen-specific T-cell responses by 'agonistic' peptides would be most desirable in response to invasive pathogenic challenges, such as infectious and neoplastic disease, while the downregulation of antigen-specific T-cell responses by 'antagonistic' peptides would be most efficacious during inappropriate pathologic consequences, such as autoimmunity. The capacity to experimentally manipulate intrinsic properties of cognate peptide ligands to appropriately alter the nature, course and potency of cellular immune interactions has important potential in both preventive and therapeutic clinical paradigms. C1 NCI, NIH, Tumor Immunol & Biol Lab, Bethesda, MD 20892 USA. RP Abrams, SI (reprint author), NCI, NIH, Tumor Immunol & Biol Lab, 10 Ctr Dr,Room 8B07,MSC, Bethesda, MD 20892 USA. NR 49 TC 19 Z9 19 U1 0 U2 0 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0952-7915 J9 CURR OPIN IMMUNOL JI Curr. Opin. Immunol. PD FEB PY 2000 VL 12 IS 1 BP 85 EP 91 DI 10.1016/S0952-7915(99)00055-2 PG 7 WC Immunology SC Immunology GA 283ZX UT WOS:000085306900012 PM 10679405 ER PT J AU Minton, AP AF Minton, AP TI Implications of macromolecular crowding for protein assembly SO CURRENT OPINION IN STRUCTURAL BIOLOGY LA English DT Review ID SELF-ASSOCIATION; EXCLUDED-VOLUME; PHYSIOLOGICAL CONSEQUENCES; POLY(ETHYLENE GLYCOL); HUMAN SPECTRIN; ACTIN; DIFFUSION; MICROTUBULES; ORGANIZATION; DETERMINANT AB Recent studies have led to increased appreciation of the influence of excluded volume in solutions of high total macromolecular content ('macromolecular crowding') upon the various classes of reaction that lead to the assembly of proteins and protein complexes. In general, crowding is expected to stabilize native protein structure relative to less compact non-native structures and to favor the formation of functional complexes of native proteins. Under certain pathological conditions, 'overcrowding' may enhance the formation of nonfunctional aggregates of non-native protein (e.g, amyloid and inclusion bodies). C1 NIDDKD, Lab Biochem & Genet, Sect Phys Biochem, NIH, Bethesda, MD 20892 USA. RP Minton, AP (reprint author), NIDDKD, Lab Biochem & Genet, Sect Phys Biochem, NIH, Bldg 8,Room 226, Bethesda, MD 20892 USA. OI Minton, Allen/0000-0001-8459-1247 NR 48 TC 418 Z9 432 U1 5 U2 54 PU CURRENT BIOLOGY LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0959-440X J9 CURR OPIN STRUC BIOL JI Curr. Opin. Struct. Biol. PD FEB PY 2000 VL 10 IS 1 BP 34 EP 39 DI 10.1016/S0959-440X(99)00045-7 PG 6 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 287WG UT WOS:000085529200004 PM 10679465 ER PT J AU Andrews, J Garcia-Estefania, D Delon, I Lu, JN Mevel-Ninio, M Spierer, A Payre, F Pauli, D Oliver, B AF Andrews, J Garcia-Estefania, D Delon, I Lu, JN Mevel-Ninio, M Spierer, A Payre, F Pauli, D Oliver, B TI OVO transcription factors function antagonistically in the Drosophila female germline SO DEVELOPMENT LA English DT Article DE sex determination; transcription; epidermis; denticle; maternal-effect sterility; maternal-effect lethality; germline; Drosophila melanogaster ID LINE SEX DETERMINATION; GENE-EXPRESSION; MELANOGASTER; LOCUS; REPRESSION; ACTIVATION; SHAVENBABY; MECHANISMS; OOGENESIS; CODONS AB OVO controls germline and epidermis differentiation in flies and mice. In the Drosophila germline, alternative OVO-B and OVO-A isoforms have a common DNA-binding domain, but different N-termini, We show that these isoforms are transcription factors with opposite regulatory activities. Using yeast one-hybrid assays, we identified a strong activation domain within a common region and a counteracting repression domain within the OVO-Aspecific region. In flies, OVO-B positively regulated the ovarian tumor promoter, while OVO-A was a negative regulator of the ovarian turner and eve promoters. OVO-B isoforms supplied ovo(+) function in the female germline and epidermis, while OVO-A isoforms had dominant-negative activity in both tissues, Moreover, elevated expression of OVO-A resulted in maternal-effect lethality while the absence of OVO-A resulted in maternal-effect sterility. Our data indicate that tight regulation of antagonistic OVO-B and OVO-A isoforms is critical for germline formation and differentiation. C1 Univ Geneva, Dept Zool & Anim Biol, Geneva, Switzerland. Ctr Dev Biol, UMR 5547, Toulouse, France. CNRS, Ctr Mol Genet, Gif Sur Yvette, France. RP Oliver, B (reprint author), NIDDK, Lab Cellular & Dev Biol, NIH, Bethesda, MD 20892 USA. NR 40 TC 34 Z9 36 U1 0 U2 3 PU COMPANY OF BIOLOGISTS LTD PI CAMBRIDGE PA BIDDER BUILDING CAMBRIDGE COMMERCIAL PARK COWLEY RD, CAMBRIDGE CB4 4DL, CAMBS, ENGLAND SN 0950-1991 J9 DEVELOPMENT JI Development PD FEB PY 2000 VL 127 IS 4 BP 881 EP 892 PG 12 WC Developmental Biology SC Developmental Biology GA 290AC UT WOS:000085653300021 PM 10648246 ER PT J AU Lilly, MA de Cuevas, M Spradling, AC AF Lilly, MA de Cuevas, M Spradling, AC TI Cyclin A associates with the fusome during germline cyst formation in the Drosophila ovary SO DEVELOPMENTAL BIOLOGY LA English DT Article DE cyclin A; cell cycle; fusome; oogenesis; Drosophila ID BAG-OF-MARBLES; CELL-CYCLE; S-PHASE; OOCYTE DIFFERENTIATION; DNA-REPLICATION; RECOMBINATION NODULES; DOWN-REGULATION; PROTEIN; MITOSIS; UBIQUITIN AB Regulated changes in the cell cycle underlie many aspects of growth and differentiation. Prior to meiosis, germ cell cycles in many organisms become accelerated, synchronized, and modified to lack cytokinesis. These changes cause cysts of interconnected germ cells to form that typically contain 2 " cells. In Drosophila, developing germ cells during this period contain a distinctive organelle, the fusome, that is required for normal cyst formation. We find that the cell cycle regulator Cyclin A transiently associates with the fusome during the cystocyte cell cycles, suggesting that fusome-associated Cyclin A drives the interconnected cells within each cyst synchronously into mitosis. In the presence of a normal fusome, overexpression of Cyclin A forces cysts through an extra round of cell division to produce cysts with 32 germline cells. Female sterile mutations in UbcD1, encoding an E2 ubiquitin-conjugating enzyme, have a similar effect. Our observations suggest that programmed changes in the expression and cytoplasmic localization of key cell cycle regulatory proteins control germline cyst production. (C) 2000 Academic Press. C1 Carnegie Inst Washington, Dept Embryol, Howard Hughes Med Inst, Baltimore, MD 21210 USA. NICHHD, Cell Biol & Metab Branch, NIH, Bethesda, MD 20892 USA. RP Lilly, MA (reprint author), Univ Maryland, HJ Patterson Hall, College Pk, MD 20742 USA. OI Lilly, Mary/0000-0003-1564-619X FU Howard Hughes Medical Institute; NIGMS NIH HHS [GM27875] NR 58 TC 52 Z9 52 U1 1 U2 8 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0012-1606 J9 DEV BIOL JI Dev. Biol. PD FEB 1 PY 2000 VL 218 IS 1 BP 53 EP 63 DI 10.1006/dbio.1999.9570 PG 11 WC Developmental Biology SC Developmental Biology GA 281RE UT WOS:000085173900005 PM 10644410 ER PT J AU Yuan, BB Li, CG Kimura, S Engelhardt, RT Smith, BR Minoo, P AF Yuan, BB Li, CG Kimura, S Engelhardt, RT Smith, BR Minoo, P TI Inhibition of distal lung morphogenesis in Nkx2.1 (-/-) embryos SO DEVELOPMENTAL DYNAMICS LA English DT Article DE lung morphogenesis; homeodomain transcription factor; Nkx2.1; TTF-1; epithelial-mesenchymal interactions; BMP4; FCF-10; collagen type IV; alpha-integrins; Vegf differential splicing; lung circulation ID MAGNETIC-RESONANCE MICROSCOPY; BRANCHING MORPHOGENESIS; CELL-DIFFERENTIATION; IN-VIVO; INTEGRIN; EXPRESSION; MOUSE; ORGANOGENESIS; EPITHELIUM; MESENCHYME AB In vitro and in vivo results are consistent with a critical role for NKX2.1, an epithelial homeodomain transcription factor in lung morphogenesis, Nkx2.1 null mutant embryos die at birth due to respiratory insufficiency caused by profoundly abnormal lungs, However, the precise role of NKX2.1 in the multistep process of lung structural morphogenesis and differentiation of various pulmonary cell types remains unknown. In the current study, we tested the hypothesis that the mutant lungs do not undergo branching morphogenesis beyond the formation of the mainstem bronchi and therefore consist solely of dilated tracheobronchial structures. To test this hypothesis, we determined the spatial and temporal expression pattern of a number of extracellular matrix (ECM) proteins and their cellular receptors, including ru-integrins, laminin, and collagen type IV. Although laminin is expressed in the mutant Nkx2.1(-/-) lungs, expression of alpha-integrins and collagen type IV is significantly reduced or absent. In addition, examination of regionally specific expression of differentially spliced Vegf (vascular endothelial growth factor) transcripts, clearly indicates that the epithelial phenotype of the Nkx2.1(-/-) lungs is similar to the tracheobronchial epithelium. In contrast to wild-type lungs in which both Vegf1 and Vegf3 are developmentally expressed, Nkx2.1(-/-) lungs are characterized by predominant expression of Vegf1 and reduced or absent Vegf3. A similar pattern of Vegf expression is also observed in isolated tracheo-bronchial tissue. The sum of these findings suggest that at least two separate pathways may exist in embryonic lung morphogenesis: proximal lung morphogenesis is Nkx2.1 independent, while distal lung morphogenesis appears to be strictly dependent on the wild-type activity of Nkx2.1. (C) 2000 Wiley-Liss, Inc. C1 Univ So Calif, Los Angeles Cty Med Ctr, Dept Pediat, Womens & Childrens Hosp,Sch Med, Los Angeles, CA 90033 USA. NCI, Lab Metab, NIH, Bethesda, MD 20892 USA. Sci Applicat Int Corp, Magnet Resonance Microscopy Grp, Frederick, MD USA. Duke Univ, Med Ctr, Dept Radiol, Durham, NC 27710 USA. RP Minoo, P (reprint author), Univ So Calif, Los Angeles Cty Med Ctr, Dept Pediat, Womens & Childrens Hosp,Sch Med, Gen Labs Bldg,1801 E Marengo St,Room 1G38, Los Angeles, CA 90033 USA. FU NHLBI NIH HHS [HL60231, HL56590] NR 37 TC 77 Z9 78 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 1058-8388 J9 DEV DYNAM JI Dev. Dyn. PD FEB PY 2000 VL 217 IS 2 BP 180 EP 190 DI 10.1002/(SICI)1097-0177(200002)217:2<180::AID-DVDY5>3.0.CO;2-3 PG 11 WC Anatomy & Morphology; Developmental Biology SC Anatomy & Morphology; Developmental Biology GA 280DB UT WOS:000085086000005 PM 10706142 ER PT J AU Resnick, HE Harris, MI Brock, DB Harris, TB AF Resnick, HE Harris, MI Brock, DB Harris, TB TI American Diabetes Association diabetes diagnostic criteria, advancing age, and cardiovascular disease risk profiles - Results from the Third National Health and Nutrition Examination Survey SO DIABETES CARE LA English DT Article ID GLUCOSE-TOLERANCE; ADULTS AB OBJECTIVE - To evaluate age-specific effects on diabetes prevalence estimates resulting from the American Diabetes Association (ADA) recommendation against use of the oral glucose tolerance rest (OGTT), we contrasted the prevalence of two mutually exclusive groups: undiagnosed diabetes according to ADA criteria (no report of diabetes and fasting glucose [FG] greater than or equal to 126 mg/dl) and isolated postchallenge hyperglycemia (IPH) (FG <126 mg/dl and OGTT greater than or equal to 200 mg/dl), a group designated to have diabetes by World Health Organization (WHO) criteria bur not ADA criteria. RESEARCH DESIGN AND METHODS - The weighted age-specific I arius of undiagnosed diabetes:IPH were calculated for 2,844 subjects aged 40-74 years without reported diabetes who had both FG and OGTT. A ratio >1.0 indicated that the proportion of undiagnosed diabetes was greater than that of IPH. Mean levels of HbA(1c) and cardiovascular disease (CVD) risk factors were contrasted among people with undiagnosed diabetes and IPH and those without either abnormality ("nondiabetic"). RESULTS - Both undiagnosed diabetes and IPH increased with age, but age-specific undiagnosed diabetes:IPH ratios decreased from 5.49 in the 40-44 age-group to 0.77 in the 70-74 age-group. Regression analysis showed a significant (P = 0.006) negative association between age and these ratios. Mean HbA(1c) was 7.1% in the undiagnosed diabetes group and differed significantly from that of the IPH and nondiabetic groups (5.6 and 5.3%, respectively). Individuals with undiagnosed diabetes had less favorable triglycerides, BMI, and HDL cholesterol compared with people with IPH. CONCLUSIONS - Compared with WHO criteria, the ADA criteria underestimate glucose abnormalities more with increasing age. However, compared to those with undiagnosed diabetes, individuals with IPH had a mean HbA(1c) level that is considered in the nondiabetic range, and this group had significantly more favorable levels of several key CVD risk factors. These findings suggest that the ADA criteria, although underestimating the abnormalities of postchallenge hyperglycemia that occur frequently with increasing age, appear to be effective at identifying a group of individuals with both unfavorable CVD risk factor profiles and evidence of long-term exposure to hyperglycemia. C1 NIA, Epidemiol Demog & Biometry Program, Bethesda, MD 20892 USA. NIDDKD, Div Endocrinol Diabet & Metab Dis, Bethesda, MD 20892 USA. RP Resnick, HE (reprint author), NIA, Epidemiol Demog & Biometry Program, 7201 Wisconsin Ave,3C-309, Bethesda, MD 20892 USA. NR 19 TC 128 Z9 137 U1 0 U2 2 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0149-5992 J9 DIABETES CARE JI Diabetes Care PD FEB PY 2000 VL 23 IS 2 BP 176 EP 180 DI 10.2337/diacare.23.2.176 PG 5 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 279HA UT WOS:000085037300009 PM 10868827 ER PT J AU Lee, ET Howard, BV Go, O Savage, PJ Fabsitz, RR Robbins, DC Welty, TK AF Lee, ET Howard, BV Go, O Savage, PJ Fabsitz, RR Robbins, DC Welty, TK TI Prevalence of undiagnosed diabetes in three American Indian populations - A comparison of the 1997 American Diabetes Association diagnostic criteria and the 1985 World Health Organization diagnostic criteria: The Strong Heart Study SO DIABETES CARE LA English DT Article ID GLUCOSE-TOLERANCE; CATEGORIES AB OBJECTIVE - In 1997, the Expert Committee on the Diagnosis and Classification of Diabetes Mellitus of the American Diabetes Association (ADA) recommended three new sets of criteria for the diagnosis of diabetes that were different from those established by the World Health Organization (WHO) in 1985. One of these three methods was based on a fasting plasma glucose value only This article compares ADA criteria with WHO criteria by applying them to three subgroups of American Indians in the Strong Heart Study who had no known diabetes. RESEARCH DESIGN AND METHODS - The Strong Heart Study is a prospective epidemiological study of vascular disease in three American Indian populations aged 45-74 ears. During the baseline examination from 1988 to 1991, participants without diagnosed diabetes underwent a fasting glucose test and a 2-h oral glucose tolerance test. These values were used to compare the ADA and WHO diagnostic criteria. RESULTS - By using fasting and 2-h glucose values, prevalence rates of undiagnosed diabetes were 15.9% according to WHO criteria and 14.4% according to ADA criteria. The overall agreement rate was 65%, and the weighted kappa statistic was 0.474, which indicates moderate agreement. The age-specific analysis showed that, among participants between 45 and 54 years of age, the prevalence rates of undiagnosed diabetes were 13.4% according to WHO criteria and 12.7% according to ADA criteria. Among those aged 55-74 years, the rates were 18.7% according to WHO criteria and 16.3% according to ADA criteria. Thus, the difference in the prevalence rates when using WHO and ADA criteria, although generally small in this population, mas three times higher in the older group (2.4%) than the difference in the younger group (0.7%). CONCLUSIONS - The Strong Heart Study found that prevalence rates of undiagnosed diabetes determined by ADA criteria and WHO criteria were similar in its American Indian population. The data suggest that the difference between the two criteria may increase as age increases. Longitudinal data will be needed to evaluate further the utility of the two criteria. C1 Univ Oklahoma, Hlth Sci Ctr, Coll Publ Hlth, Ctr Amer Indian Hlth Res, Oklahoma City, OK 73190 USA. Medlant Res Inst, Washington, DC USA. NHLBI, NIH, Bethesda, MD 20892 USA. Aberdeen Area Tribal Chairmens Hlth Board, Rapid City, SD USA. RP Lee, ET (reprint author), Univ Oklahoma, Hlth Sci Ctr, Coll Publ Hlth, Ctr Amer Indian Hlth Res, POB 26901, Oklahoma City, OK 73190 USA. FU NHLBI NIH HHS [U01-HL-41654, U01-HL-41652, U01-HL-41642] NR 13 TC 29 Z9 30 U1 0 U2 0 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0149-5992 J9 DIABETES CARE JI Diabetes Care PD FEB PY 2000 VL 23 IS 2 BP 181 EP 186 DI 10.2337/diacare.23.2.181 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 279HA UT WOS:000085037300010 PM 10868828 ER PT J AU Rohlfing, CL Little, RR Wiedmeyer, HM England, JD Madsen, R Harris, MI Flegal, KM Eberhardt, MS Goldstein, DE AF Rohlfing, CL Little, RR Wiedmeyer, HM England, JD Madsen, R Harris, MI Flegal, KM Eberhardt, MS Goldstein, DE TI Use of GHb (HbA(1c)) in screening for undiagnosed diabetes in che US population SO DIABETES CARE LA English DT Article ID GLUCOSE-TOLERANCE TEST; FASTING PLASMA-GLUCOSE; GLYCOSYLATED HEMOGLOBIN; MICROVASCULAR COMPLICATIONS; GLYCATED HEMOGLOBIN; MELLITUS; DIAGNOSIS; INTOLERANCE; FRUCTOSAMINE AB OBJECTIVE - To evaluate the use of GHb as a screening test for undiagnosed diabetes (fasting plasma glucose greater than or equal to 7.0 mmol/l) in a representative sample of the U.S. population. RESEARCH DESIGN AND METHODS - The Third National Health and Nutrition Examination Sun-ey included national samples of non-Hispanic whites, non-Hispanic blacks, and Mexican Americans aged greater than or equal to 20 years. Of these subjects, 7,832 participated in a morning examination session, of which 1,273 were excluded because of a previous diagnosis of diabetes, missing data, or fasting rime of <8 h before examination. Venous blood was obtained to measure fasting plasma glucose and GHb in the remaining 6,559 subjects. Received operating characteristic curve analysis was used To examine the sensitivity and specificity of GHb for detecting diabetes at increasing GHb cutoff levels. RESULTS - GHb demonstrated high sensitivity (83.4%) and specificity(83.4%) for detecting undiagnosed diabetes at a GHb cutoff of 1 SD above the normal mean. Moderate sensitivity (63.2%) and very high specificity (97.4%) were evident at a GHb cutoff of 2 SD above the normal mean, Sensitivity at this level ranged from 58.6% in the non-Hispanic white population to 83.6% in the Mexican-American population; specificity ranged from 93.0% in the non-Hispanic black population to 98.3% in the non-Hispanic white population. CONCLUSIONS - GHb is a highly specific and convenient alternative to fasting plasma glucose for diabetes screening. A GHb value of 2 SD above the normal mean could identify a high proportion of individuals with undiagnosed diabetes who are at risk for developing diabetes complications. C1 Univ Missouri, BES, Dept Child Hlth, Sch Med, Columbia, MO 65203 USA. NIDDKD, NIH, Bethesda, MD 20892 USA. Natl Ctr Hlth Stat, Hyattsville, MD 20782 USA. RP Rohlfing, CL (reprint author), Univ Missouri, BES, Dept Child Hlth, Sch Med, 1 Hosp Dr,M772, Columbia, MO 65203 USA. RI Flegal, Katherine/A-4608-2013; OI Flegal, Katherine/0000-0002-0838-469X; Little, Randie/0000-0001-6450-8012 NR 40 TC 232 Z9 244 U1 0 U2 3 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0149-5992 J9 DIABETES CARE JI Diabetes Care PD FEB PY 2000 VL 23 IS 2 BP 187 EP 191 DI 10.2337/diacare.23.2.187 PG 5 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 279HA UT WOS:000085037300011 PM 10868829 ER PT J AU Hawa, MI Fava, D Medici, F Deng, YJ Notkins, AL De Mattia, G Leslie, RDG AF Hawa, MI Fava, D Medici, F Deng, YJ Notkins, AL De Mattia, G Leslie, RDG TI Antibodies to IA-2 and GAD65 in type 1 and type 2 diabetes - Isotype restriction and polyclonality SO DIABETES CARE LA English DT Article ID GLUTAMIC-ACID DECARBOXYLASE; IGG SUBCLASS DISTRIBUTION; MONOCLONAL-ANTIBODIES; FOLLOW-UP; INSULIN; AUTOANTIBODIES; PREDICTION; PATIENT; DISEASE; MELLITUS AB OBJECTIVE - To determine the isotypes and clonality of antibodies to GAD (GADA) and 1A-2 (1A-2A) in patients with type 1 and type 2 diabetes. RESEARCH DESIGN AND METHODS - We studied the following consecutive series of patients who attended a diabetes center for antibodies to GADA and 1A-2A: 52 newly diagnosed type 1 diabetic patients, 199 type 2 diabetic patients, 200 control patients, and a cohort of 34 nondiabetic identical twins of patients with type 1 diabetes (15 of whom developed diabetes) who were followed prospectively RESULTS- GADA or 1A-2A were detected in 37 (71%) type 1 diabetic patients compared with only 10 (5%) type 2 diabetic patients (P < 0.0001). Both GAD and 1A-2 antibodies, regardless of the type of diabetes, were usually subclass restricted to IgG1 and were polyclonal. IgM, IgG3, and IgE isotypes were also detected, but all isotypes of GADA and 1A-2A were less prevalent than IgG 1 (P < 0.017 for either antibody), There was no evidence of spreading or switching of isotypes before the onset of type 1 diabetes. CONCLUSIONS - These observations suggest that the pathogenesis of antigen-specific antibodies in type 1 and type 2 diabetes is similar and probably involves a chronic nonrandom antigen-driven polyclonal B-cell activation that is consistent with a Th1-type immune response. C1 St Bartholomews Hosp, Dept Diabet & Metab, London EC1A 7BE, England. Univ La Sapienza, Med Clin 1, Fdn A Cesalpino, Rome, Italy. NIH, Oral Med Lab, Bethesda, MD 20892 USA. RP Leslie, RDG (reprint author), St Bartholomews Hosp, Dept Diabet, London EC1A 7BE, England. NR 28 TC 56 Z9 62 U1 1 U2 2 PU AMER DIABETES ASSOC PI ALEXANDRIA PA 1660 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0149-5992 J9 DIABETES CARE JI Diabetes Care PD FEB PY 2000 VL 23 IS 2 BP 228 EP 233 DI 10.2337/diacare.23.2.228 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 279HA UT WOS:000085037300018 PM 10868836 ER PT J AU Robles, E Silverman, K Preston, KL Cone, EJ Katz, E Bigelow, GE Stitzer, ML AF Robles, E Silverman, K Preston, KL Cone, EJ Katz, E Bigelow, GE Stitzer, ML TI The brief abstinence test: voucher-based reinforcement of cocaine abstinence SO DRUG AND ALCOHOL DEPENDENCE LA English DT Article DE voucher-based reinforcement; abstinence initiation; cocaine abuse; urinalysis; methadone patients; contingency contracting ID CARBON-MONOXIDE REDUCTION; METHADONE PATIENTS; PAY AMOUNT; CRACK USE; INCENTIVES; CLIENTS AB This study assessed the effectiveness of a brief abstinence reinforcement procedure for initiating cocaine abstinence in methadone maintenance patients. On Monday of the test week, 72 cocaine-abusing methadone patients were offered a $100 voucher if urine samples collected on Wednesday indicated that they had abstained from cocaine across that 2-day period. A patient was considered abstinent and the voucher delivered if the urine benzoylecgonine concentration decreased by 50% from Monday to Wednesday (quantitative criterion) or if the concentration of Wednesday's urine sample was less than or equal to 300 ng/ml. Overall, 79% of study patients showed urinalysis evidence of abstention from cocaine between Monday and Wednesday of the test week. In a subsample with complete data (n = 50), significantly more patients abstained from cocaine from Monday to Wednesday of the test week (84%) than from Monday to Wednesday of the week before (36%) or after (32%) the test week. Furthermore, while almost all patients (94%) decreased their benzoylecgonine concentration from Monday to Wednesday of the test week, significantly fewer patients' benzoylecgonine concentrations decreased from Monday to Wednesday of the week before (56%) or after (48%) the test week. This highly efficacious procedure may have clinical application where reliable abstinence initiation is desired, either on a temporary basis (e.g. sobriety sampling) or at the start of longer-term interventions. It may also be possible to use the brief abstinence test as an experimental model to assess the effects of other therapeutic interventions on abstinence initiation in treatment settings. (C) 2000 Elsevier Science Ireland Ltd. All rights reserved. C1 Johns Hopkins Univ, Sch Med, Bayview Med Ctr, Behav Pharmacol Res Unit, Baltimore, MD 21224 USA. Univ Arkansas Med Sci, Little Rock, AR 72205 USA. NIDA, Intramural Res Program, Baltimore, MD 21224 USA. RP Stitzer, ML (reprint author), Johns Hopkins Univ, Sch Med, Bayview Med Ctr, Behav Pharmacol Res Unit, 5510 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Preston, Kenzie/J-5830-2013; OI Preston, Kenzie/0000-0003-0603-2479; Silverman, Kenneth/0000-0003-2724-1413 FU NIDA NIH HHS [K05 DA00050, P50 DA09258, T32 DA07209] NR 27 TC 35 Z9 35 U1 0 U2 1 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 FEB 1 PY 2000 VL 58 IS 1-2 BP 205 EP 212 DI 10.1016/S0376-8716(99)00090-3 PG 8 WC Substance Abuse; Psychiatry SC Substance Abuse; Psychiatry GA 271BP UT WOS:000084573100025 PM 10669073 ER PT J AU Ravn, P Kjaer, S Jensen, KH Wind, T Jensen, KB Kristensen, P Brosh, RM Orren, DK Bohr, VA Clark, BFC AF Ravn, P Kjaer, S Jensen, KH Wind, T Jensen, KB Kristensen, P Brosh, RM Orren, DK Bohr, VA Clark, BFC TI Identification of phage antibodies toward the Werner protein by selection on Western blots SO ELECTROPHORESIS LA English DT Article DE phage antibody; selection strategy; Western blot; human Werner protein; recombinant human Werner protein ID HUMAN MONOCLONAL-ANTIBODIES; SINGLE-CHAIN FV; DISPLAY LIBRARY; FILAMENTOUS BACTERIOPHAGES; FRAGMENTS; AFFINITY; CELLS; GENE; PURIFICATION; EXPRESSION AB A procedure was established for selecting phage antibodies (phage-abs) from phage-displayed antibody repertoires by panning against proteins, separated by sodium dodecyl phosphate-polyacrylamide gel electrophoresis (SDS-PAGE) and electroblotted onto nitrocellulose membranes (Western blots). This immobilization strategy is applicable for secondary rounds of panning in selections against semipurified proteins, and directs the selection toward antibodies suitable as immunochemical reagents in Western blots. In model experiments, enrichment factors as high as 1.9 x 10(5) were obtained in a single round of panning. Furthermore, we demonstrate the application of this approach by selection of phage-abs recognizing the human Werner protein, which is defective in a premature aging syndrome. C1 Aarhus Univ, Dept Mol & Struct Biol, DK-8000 Aarhus C, Denmark. Karolinska Inst, Dept Neurosci, Stockholm, Sweden. NIA, Genet Mol Lab, NIH, Baltimore, MD 21224 USA. RP Ravn, P (reprint author), Aarhus Univ, Dept Mol & Struct Biol, Gustav Wieds Vej 10 C, DK-8000 Aarhus C, Denmark. RI Jensen, Kim/F-5333-2014; OI Jensen, Kim/0000-0001-6569-1664; Kristensen, Peter/0000-0001-7205-6853 NR 34 TC 8 Z9 8 U1 0 U2 4 PU WILEY-V C H VERLAG GMBH PI BERLIN PA MUHLENSTRASSE 33-34, D-13187 BERLIN, GERMANY SN 0173-0835 J9 ELECTROPHORESIS JI Electrophoresis PD FEB PY 2000 VL 21 IS 3 BP 509 EP 516 DI 10.1002/(SICI)1522-2683(20000201)21:3<509::AID-ELPS509>3.3.CO;2-X PG 8 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 291HN UT WOS:000085728600005 PM 10726750 ER PT J AU Bergman, AC Benjamin, T Alaiya, A Waltham, M Sakaguchi, K Franzen, B Linder, S Bergman, T Auer, G Appella, E Wirth, PJ Jornvall, H AF Bergman, AC Benjamin, T Alaiya, A Waltham, M Sakaguchi, K Franzen, B Linder, S Bergman, T Auer, G Appella, E Wirth, PJ Jornvall, H TI Identification of gel-separated tumor marker proteins by mass spectrometry SO ELECTROPHORESIS LA English DT Article DE two-dimensional gel electrophoresis; mass spectrometry; tumor markers ID POLYPEPTIDE EXPRESSION; MALIGNANT-TUMORS; HUMAN LIVER; ELECTROPHORESIS; GALECTIN-1; CARCINOMA; PEPTIDES; BENIGN; CELLS AB Two-dimensional gel electrophoresis with subsequent analysis by mass spectrometry was applied to study differences in protein expression between benign and malignant solid tumors from human beast, lung and ovary cells. Cells from freshly resected clinical material were lysed and the extracts were subjected to isoelectric focusing with immobilized pH gradients followed by second-dimensional separation on 10-13% sodium dodecyl sulfate (SDS)/polyacrylamide gels. Polypeptides were identified using matrix-assisted laser desorption/ionization and electrospray ionization mass spectrometry after in-gel protein digestion. Some of the upregulated polypeptides in malignant cells are of potential importance as markers of tumor proliferation. Twenty such proteins were identified, ten constituting novel identifications and ten sequence verifications of previously gel-matched proteins. The proteins identified span a wide range of functions, but several cases of protein truncation were found. Truncated forms of cytokeratins 6D and 8, and of cathepsin D were identified. Truncated froms of these overexpressed proteins support the presence of proteolytic processing steps in tumor material. The protein processing and the difference between protein and mRNA abundancies in tumors of different malignancy and origin suggest that studies at the protein level are important for an understanding of tumor phenotypes. C1 Karolinska Inst, Dept Med Biochem & Biophys, S-17177 Stockholm, Sweden. NCI, Expt Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. Karolinska Hosp & Inst, Dept Pathol & Oncol, Stockholm, Sweden. NCI, Mol Pharmacol Lab, NIH, Bethesda, MD 20892 USA. RP Jornvall, H (reprint author), Karolinska Inst, Dept Med Biochem & Biophys, S-17177 Stockholm, Sweden. RI Waltham, Mark/A-1728-2009 NR 23 TC 74 Z9 89 U1 0 U2 6 PU WILEY-V C H VERLAG GMBH PI BERLIN PA MUHLENSTRASSE 33-34, D-13187 BERLIN, GERMANY SN 0173-0835 J9 ELECTROPHORESIS JI Electrophoresis PD FEB PY 2000 VL 21 IS 3 BP 679 EP 686 DI 10.1002/(SICI)1522-2683(20000201)21:3<679::AID-ELPS679>3.3.CO;2-1 PG 8 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 291HN UT WOS:000085728600032 PM 10726777 ER PT J AU Vajo, Z Francomano, CA Wilkin, DJ AF Vajo, Z Francomano, CA Wilkin, DJ TI The molecular and genetic basis of fibroblast growth factor receptor 3 disorders: The achondroplasia family of skeletal dysplasias, Muenke craniosynostosis, and Crouzon syndrome with acanthosis nigricans SO ENDOCRINE REVIEWS LA English DT Review ID JACKSON-WEISS-SYNDROME; AUTOSOMAL-DOMINANT CRANIOSYNOSTOSIS; RECURRENT G380R MUTATIONS; TYROSINE KINASE DOMAIN; THANATOPHORIC DYSPLASIA; PFEIFFER-SYNDROME; POINT MUTATION; FGFR3 GENE; TRANSMEMBRANE DOMAIN; APERT-SYNDROME AB Achondroplasia, the most common form of short-limbed dwarfism in humans, occurs between 1 in 15,000 and 40,000 Live births. More than 90% of cases are sporadic and there is, on average, an increased paternal age at the time of conception of affected individuals. More then 97% of persons with achondroplasia have a Gly380Arg mutation in the transmembrane domain of the fibroblast growth factor receptor (FGFR) 3 gene. Mutations in the FGFR3 gene also result in hypochondroplasia, the lethal thanatophoric dysplasias, the recently described SADDAN (severe achondroplasia with developmental delay and acanthosis nigricans) dysplasia, and two craniosynostosis disorders: Muenke coronal craniosynostosis and Crouzon syndrome with acanthosis nigricans. Recent evidence suggests that the phenotypic differences may be due to specific alleles with varying degrees of ligand-independent activation, allowing the receptor to be constitutively active. Since the Gly380Arg achondroplasia mutation was recognized, similar observations regarding the conserved nature of FGFR mutations and resulting phenotype have been made regarding other skeletal phenotypes, including hypochondroplasia, thanatophoric dysplasia, and Muenke coronal craniosynostosis. These specific genotype-phenotype correlations in the FGFR disorders seem to be unprecedented in the study of human disease. The explanation for this high degree of mutability at specific bases remains an intriguing question. C1 NIDCR, Craniofacial & Skeletal Dis Branch, NIH, Bethesda, MD 20892 USA. Vet Affairs Med Ctr, Dept Endocrinol & Med, Phoenix, AZ 85012 USA. Natl Human Genome Res Inst, Med Genet Branch, Bethesda, MD 20892 USA. RP Wilkin, DJ (reprint author), NIDCR, Craniofacial & Skeletal Dis Branch, NIH, 30 Convent Dr,Bldg 30,Room 228, Bethesda, MD 20892 USA. EM dwilkin@dir.nidcr.nih.gov NR 149 TC 191 Z9 205 U1 1 U2 10 PU ENDOCRINE SOC PI CHEVY CHASE PA 8401 CONNECTICUT AVE, SUITE 900, CHEVY CHASE, MD 20815-5817 USA SN 0163-769X EI 1945-7189 J9 ENDOCR REV JI Endocr. Rev. PD FEB PY 2000 VL 21 IS 1 BP 23 EP 39 DI 10.1210/er.21.1.23 PG 17 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 304YT UT WOS:000086512800003 PM 10696568 ER PT J AU Mann, CL Hughes, FM Cidlowski, JA AF Mann, CL Hughes, FM Cidlowski, JA TI Delineation of the signaling pathways involved in glucocorticoid-induced and spontaneous apoptosis of rat thymocytes SO ENDOCRINOLOGY LA English DT Article ID CELL-DEATH; TRANSFORMING GROWTH-FACTOR-BETA-1; DEOXYRIBONUCLEIC-ACID; PROTEASE ACTIVATION; REGRESSING LIVER; DNA-DEGRADATION; LYMPHOCYTES; THYMUS; INHIBITOR; FRAGMENTATION AB In primary rat thymocytes, both glucocorticoids and the withdrawal of in vivo survival factors elicit apoptosis. In this study we wanted to determine whether distinct pathways leading to apoptosis are engaged by these two stimuli. To address this question, we conducted a multiparametric analysis of cell viability, DNA fragmentation, activation of caspase-3-like activity, cell shrinkage, the loss of mitochondrial membrane potential, and externalization of phosphatidylserine in the absence and presence of protein and RNA synthesis. The role of caspase activity was also examined in both glucocorticoid- and survival factor withdrawal-induced cell death. We show that glucocorticoid-induced, but not spontaneous, loss of viability is dependent upon macromolecular synthesis and caspase activity. Furthermore, glucocorticoid-induced phosphatidylserine externalization and cell shrinkage are dependent upon gene regulation and caspase activity, whereas these features manifest independently of gene regulation and caspase activity in spontaneous death. In contrast, the loss of mitochondrial membrane potential was dependent upon macromolecular synthesis only in glucocorticoid-induced death and was independent of caspases in both spontaneous and dexamethasone-induced death. These results suggest that thymocytes can die by a caspase-independent mechanism and that a major difference between glucocorticoid- and survival factor deprivation-induced death is the dependence on gene expression. C1 NIEHS, Lab Signal Transduct, NIH, Res Triangle Pk, NC 27709 USA. Univ N Carolina, Curriculum Toxicol, Chapel Hill, NC 27599 USA. RP Cidlowski, JA (reprint author), POB 12233,MD E2-02, Res Triangle Pk, NC 27709 USA. EM cidlowski@niehs.nih.gov NR 42 TC 51 Z9 52 U1 0 U2 3 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 FEB PY 2000 VL 141 IS 2 BP 528 EP 538 DI 10.1210/en.141.2.528 PG 11 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 276ET UT WOS:000084863200008 PM 10650932 ER PT J AU Greenland, KJ Jantke, I Jenatschke, S Bracken, KE Vinson, C Gellersen, B AF Greenland, KJ Jantke, I Jenatschke, S Bracken, KE Vinson, C Gellersen, B TI The human NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase gene promoter is controlled by Ets and activating protein-1 transcription factors and progesterone SO ENDOCRINOLOGY LA English DT Article ID ENDOMETRIAL STROMAL CELLS; PROSTAGLANDIN DEHYDROGENASE; 11-BETA-HYDROXYSTEROID DEHYDROGENASE; PROLACTIN GENE; KINASE-A; 15-HYDROXY-PROSTAGLANDIN DEHYDROGENASE; GLUCOCORTICOID-RECEPTOR; DEPENDENT TRANSCRIPTION; PROSTANOID RECEPTORS; MIFEPRISTONE RU486 AB NAD(+)-dependent 15-hydroxyprostaglandin dehydrogenase (PGDH) is a key catabolic enzyme in the inactivation of PGF(2 alpha) and PGE, and therefore serves as an important determinant in regulating their local concentrations. To gain insights into the transcriptional regulation of this enzyme, we have isolated 3.5 kb of the 5'-flanking sequence of the human PGDH promoter and characterized its control in hemopoietic cells and cells of myometrial and placental origin. Several potential binding sites for cAMP-responsive element-binding protein (CREB), Ets, and activating protein-1 (AP-1) transcription factors are present within 2368 bp of the 5'-flanking region. This region and deletions thereof were fused to the luciferase reporter gene and used for transient transfection experiments. In Jurkat leukemic T cells, which express PGDH endogenously, the transfected PGDH promoter was strongly induced by phorbol ester. Induction was reversed by coexpression of A-Fos, a dominant negative to AP-1. In primary cultures of myometrial smooth muscle cells (SMC), the Ets family members Ets-1, Ets-2, and PEA3 potently stimulated transcriptional activity of the PGDH promoter. PEAS-mediated activation was partially repressed by A-Fos, suggesting an involvement of AP-1 proteins, which might be conferred by a distal and a proximal Ets/ AP-1 composite element. The distal Ets/AP-1 element is flanked by two CRE-like sequences. Cotransfection of A-CREB, a dominant negative to CREB, inhibited stimulation of PGDH-2368/luc3 by PEA3 in myometrial SMC, whereas treatment with 8-bromo-cAMP moderately enhanced promoter activity. Progesterone is believed to be an important stimulus for PGDH expression in the utero-placental unit, thus contributing to the maintenance of a quiescent uterus during pregnancy. In myometrial SMC, both isoforms of the progesterone receptor, PR-B and PR-A, caused a ligand-dependent activation of PGDH-2368/luc3. Transcriptional activity of PR-B, but not PR-A, was further enhanced by the addition of 8-bromo-cAMP. We could not confirm a recently proposed transcriptional control of PGDH by mineralocorticoid receptor. No effect of mineralocorticoid receptor, in the absence or presence of aldosterone, with or without 8-bromo-cAMP, was observed on PGDH-2368/luc3. Taken together, these findings demonstrate control of the PGDH promoter by multiple pathways and provide evidence for cross-talk among Ets, AP-1, cAMP, and PR-mediated signaling, suggesting complex regulatory mechanisms for the expression of PGDH. C1 Univ Hamburg, IHF Inst Hormone & Fertil Res, D-22529 Hamburg, Germany. NCI, Biochem Lab, NIH, Bethesda, MD 20892 USA. RP Gellersen, B (reprint author), Univ Hamburg, IHF Inst Hormone & Fertil Res, Grandweg 64, D-22529 Hamburg, Germany. EM gellersen@ihf.de NR 81 TC 62 Z9 62 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 FEB PY 2000 VL 141 IS 2 BP 581 EP 597 DI 10.1210/en.141.2.581 PG 17 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 276ET UT WOS:000084863200015 PM 10650939 ER PT J AU Royaux, IE Suzuki, K Mori, A Katoh, R Everett, LA Kohn, LD Green, ED AF Royaux, IE Suzuki, K Mori, A Katoh, R Everett, LA Kohn, LD Green, ED TI Pendrin, the protein encoded by the Pendred syndrome gene (PDS), is an apical porter of iodide in the thyroid and is regulated by thyroglobulin in FRTL-5 cells SO ENDOCRINOLOGY LA English DT Article ID FOLLICULAR THYROGLOBULIN; SULFATE TRANSPORTER; NA+/I-SYMPORTER; PLASMA-MEMBRANE; HEARING-LOSS; EXPRESSION; TRANSCRIPTION; CLONING; IDENTIFICATION; HETEROGENEITY AB Pendred syndrome is an autosomal recessive disorder characterized by congenital deafness and thyroid goiter. The thyroid disease typically develops around puberty and is associated with a mild organification defect, characterized by an inappropriate discharge of iodide upon perchlorate stimulation (a positive perchlorate discharge test). The gene (PDS) mutated in Pendred syndrome is expressed in thyroid and encodes a 780-amino acid protein (pendrin) that has recently been shown to function as an iodide/chloride transporter. We sought to establish the location of pendrin in the thyroid and to examine the regulatory network controlling its synthesis. Using peptide-specific antibodies for immunolocalization studies, pendrin was detected in a limited subset of cells within the thyroid follicles, exclusively at the apical membrane of the follicular epithelium. Interestingly, significantly greater amounts of pendrin were encountered in thyroid tissue from patients with Graves' disease. Using a cultured rat thyroid cell line (FRTL-5), PDS expression was found to be significantly induced by low concentrations of thyroglobulin (TG), but not by TSH, sodium iodide, or insulin. This is different from the established effect of TG, more typically a potent suppressor of thyroid-specific gene expression. Together, these results suggest that pendrin is an apical porter of iodide in the thyroid and that the expression and function of both the apical and basal iodide porters are coordinately regulated by follicular TG. C1 Natl Human Genome Res Inst, Genome Technol Branch, NIH, Bethesda, MD 20892 USA. NIDDKD, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. Yamanashi Med Univ, Sch Med, Dept Pathol, Tamaho, Yamanashi 40938, Japan. RP Green, ED (reprint author), Natl Human Genome Res Inst, Genome Technol Branch, NIH, 49 Convent Dr,Bldg 49,Room 2A08, Bethesda, MD 20892 USA. EM egreen@nhgri.nih.gov NR 42 TC 259 Z9 278 U1 0 U2 6 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 FEB PY 2000 VL 141 IS 2 BP 839 EP 845 DI 10.1210/en.141.2.839 PG 7 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 276ET UT WOS:000084863200043 PM 10650967 ER PT J AU van Birgelen, APJM AF van Birgelen, APJM TI Hexachlorobenzene: van Birgelen's response SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Letter C1 NIEHS, Res Triangle Pk, NC 27709 USA. RP van Birgelen, APJM (reprint author), NIEHS, POB 12233, Res Triangle Pk, NC 27709 USA. NR 3 TC 0 Z9 0 U1 0 U2 1 PU US DEPT HEALTH HUMAN SCIENCES PUBLIC HEALTH SCIENCE PI RES TRIANGLE PK PA NATL INST HEALTH, NATL INST ENVIRONMENTAL HEALTH SCIENCES, PO BOX 12233, RES TRIANGLE PK, NC 27709-2233 USA SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD FEB PY 2000 VL 108 IS 2 BP A58 EP A58 PG 1 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 287LY UT WOS:000085508400003 ER PT J AU Wilson, SH AF Wilson, SH TI Environmental medicine at a crossroad: Health in the United States SO ENVIRONMENTAL HEALTH PERSPECTIVES LA English DT Editorial Material ID LEAD C1 NIEHS, Res Triangle Pk, NC 27709 USA. RP Wilson, SH (reprint author), NIEHS, POB 12233, Res Triangle Pk, NC 27709 USA. NR 9 TC 1 Z9 1 U1 0 U2 0 PU US DEPT HEALTH HUMAN SCIENCES PUBLIC HEALTH SCIENCE PI RES TRIANGLE PK PA NATL INST HEALTH, NATL INST ENVIRONMENTAL HEALTH SCIENCES, PO BOX 12233, RES TRIANGLE PK, NC 27709-2233 USA SN 0091-6765 J9 ENVIRON HEALTH PERSP JI Environ. Health Perspect. PD FEB PY 2000 VL 108 IS 2 BP A56 EP A56 PG 1 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 287LY UT WOS:000085508400001 PM 10656861 ER PT J AU Bottner, A Eisenhofer, G Friberg, P Rundqvist, B Bornstein, SR AF Bottner, A Eisenhofer, G Friberg, P Rundqvist, B Bornstein, SR TI Serum leptin levels in heart failure patients may be altered differently according to clinical stage SO EUROPEAN HEART JOURNAL LA English DT Letter C1 Univ Leipzig, Dept Internal Med 3, D-7010 Leipzig, Germany. NINDS, Clin Neurosci Branch, Bethesda, MD 20892 USA. Gothenburg Univ, Dept Clin Physiol, S-41124 Gothenburg, Sweden. NICHHD, Bethesda, MD 20892 USA. RP Bottner, A (reprint author), Univ Leipzig, Dept Internal Med 3, D-7010 Leipzig, Germany. RI Korner, Antje/B-3988-2015 OI Korner, Antje/0000-0001-6001-0356 NR 9 TC 3 Z9 3 U1 0 U2 0 PU W B SAUNDERS CO LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0195-668X J9 EUR HEART J JI Eur. Heart J. PD FEB PY 2000 VL 21 IS 4 BP 334 EP 335 DI 10.1053/euhj.1999.1979 PG 2 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 284PM UT WOS:000085341000017 ER PT J AU Lener, M Horn, IR Cardinale, A Messina, S Nielsen, UB Rybak, SM Hoogenboom, HR Cattaneo, A Biocca, S AF Lener, M Horn, IR Cardinale, A Messina, S Nielsen, UB Rybak, SM Hoogenboom, HR Cattaneo, A Biocca, S TI Diverting a protein from its cellular location by intracellular antibodies - The case of p21Ras SO EUROPEAN JOURNAL OF BIOCHEMISTRY LA English DT Article DE intracellular antibodies; scFv fragments; anti-p21Ras; phage display library ID SINGLE-CHAIN ANTIBODY; MONOCLONAL-ANTIBODIES; ONCOGENE PRODUCT; MAMMALIAN-CELLS; PHAGE DISPLAY; RAS; EXPRESSION; VIRUS; IMMUNIZATION; FRAGMENTS AB We describe the use of phage libraries to derive new antibodies against p21Ras to be used for intracellular expression in mammalian cells. A panel of single-chain antibody fragments, binding to Ras, were analyzed and characterized for their capacity to interfere in vitro with (a) the intrinsic GTPase activity of Ras and (b) the binding of Ras to its effector Raf, and were found not to neutralize its function, according to these biochemical criteria. When expressed intracellularly in mouse 3T3 K-Ras transformed cells all the anti-aas single-chain variable fragments (scFv) tested inhibited cell proliferation, as assessed by bromodeoxyuridine incorporation. Double immunofluorescence analysis of transfected cells using confocal microscopy confirmed that anti-Ras antibody fragments colocalize with endogenous Ras, at subcellular locations where the protein Ras is not normally found. These data suggest that the ability of phage-derived anti-Ras scFv fragments to inhibit the function of Ras in vivo is a rather general and frequent property and that the range of antibodies that can be successfully used for intracellular inhibition studies is much greater than anticipated, exploiting the mode of action of diverting protein traffic. C1 Univ Roma Tor Vergata, Dept Neurosci, I-00133 Rome, Italy. Univ Hosp Maastricht, Dept Pathol, Maastricht, Netherlands. NCI, Dev Therapeut Program, FCRDC, NIH, Frederick, MD 21701 USA. Int Sch Adv Studies, Program Neurosci, Trieste, Italy. RP Biocca, S (reprint author), Univ Roma Tor Vergata, Dept Neurosci, Via Tor Vergata 135, I-00133 Rome, Italy. OI MESSINA, Samantha/0000-0003-2185-8236; CATTANEO, ANTONINO/0000-0002-6975-8923 NR 50 TC 55 Z9 56 U1 1 U2 2 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0014-2956 J9 EUR J BIOCHEM JI Eur. J. Biochem. PD FEB PY 2000 VL 267 IS 4 BP 1196 EP 1205 DI 10.1046/j.1432-1327.2000.01125.x PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 291MG UT WOS:000085738300034 PM 10672031 ER PT J AU Castagne, C Murphy, EC Gronenborn, AM Delepierre, M AF Castagne, C Murphy, EC Gronenborn, AM Delepierre, M TI P-31 NMR analysis of the DNA conformation induced by protein binding - SRY/DNA complexes SO EUROPEAN JOURNAL OF BIOCHEMISTRY LA English DT Article DE P-31-NMR; SRY protein; DNA-protein complex ID DETERMINING FACTOR SRY; NUCLEIC-ACIDS; SEX DETERMINATION; MOLECULAR-BASIS; SPECTROSCOPY; ANGLES; SITE; H-1; RECOGNITION; DEFINITION AB Complexes of the HMG box protein SRY with two duplexes of 8 and 14 base pairs have been studied by P-31 NMR and complete assignment of all phosphorus signals of the bound DNA duplexes are presented. While for the free DNA, all P-31 signals display Limited spectral dispersion (< 0.8 p.p.m.) for the bound duplexes, P-31 resonances are spread over 2 p.p.m. Based on the previously published 3D structure of hSRY-HMG, with the 8 mer it is demonstrated that the upfield shifted resonances correspond to the site of partial intercalation of an isoleucine side chain into the DNA. Moreover, the observation of significant difference in linewidths between the two duplexes allows to estimate lifetime of the complexes from P-31-P-31 2D exchange experiments. C1 Inst Pasteur, Nucl Magnet Resonance Lab, CNRS, URA 1129, F-75724 Paris 15, France. NIDDKD, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. RP Delepierre, M (reprint author), Inst Pasteur, Nucl Magnet Resonance Lab, CNRS, URA 1129, 28 Rue Dr Roux, F-75724 Paris 15, France. NR 41 TC 16 Z9 16 U1 0 U2 5 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0014-2956 J9 EUR J BIOCHEM JI Eur. J. Biochem. PD FEB PY 2000 VL 267 IS 4 BP 1223 EP 1229 DI 10.1046/j.1432-1327.2000.01124.x PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 291MG UT WOS:000085738300037 PM 10672034 ER PT J AU Chan, FKM Lenardo, MJ AF Chan, FKM Lenardo, MJ TI A crucial role for p80 TNF-R2 in amplifying p60 TNF-R1 apoptosis signals in T lymphocytes SO EUROPEAN JOURNAL OF IMMUNOLOGY LA English DT Article DE TNF-alpha; NF-kappa B; TRAF2; signal transduction ID TUMOR-NECROSIS-FACTOR; NF-KAPPA-B; DOMAIN-CONTAINING RECEPTOR; INDUCED CELL-DEATH; DEPENDENT APOPTOSIS; KINASE RIP; ACTIVATION; COMPLEX; PROTEIN; FADD AB Tumor necrosis factor-alpha (TNF-alpha) can elicit many cellular responses including programmed cell death or apotosis. TNF-alpha-induced apoptosis has been largely attributed to the p60 TNF-R1 receptor The role of p80 in TNF-alpha-mediated apoptosis is largely unknown. We now present evidence that signaling through p80 switches on the previously dormant apoptotic machinery associated with p60. This effect on p60-associated apoptosis involves the proximal activation of caspases and proceeds in the presence of strong NF-kappa B induction. We evaluated the role of TRAF2 in p80-assisted apoptosis and found that a decrease in TRAF2 protein occurs with p80 but not p60 stimulation. However, the decrease in TRAF2 protein can be dissociated from apoptosis in the presence of a caspase inhibitor. Hence, one means by which p80 TNF-R2 regulates apoptosis is through its ability to amplify internally apoptotic signal transduction from p60 TNF-R1. C1 NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Lenardo, MJ (reprint author), NIAID, Immunol Lab, NIH, Bldg 10,Room 11N311,10 Ctr Dr,MSC-1892, Bethesda, MD 20892 USA. RI Chan, Francis/E-9647-2014; OI Chan, Francis/0000-0002-4803-8353; Chan, Francis K.L./0000-0001-7388-2436 NR 42 TC 105 Z9 108 U1 0 U2 3 PU WILEY-V C H VERLAG GMBH PI BERLIN PA MUHLENSTRASSE 33-34, D-13187 BERLIN, GERMANY SN 0014-2980 J9 EUR J IMMUNOL JI Eur. J. Immunol. PD FEB PY 2000 VL 30 IS 2 BP 652 EP 660 PG 9 WC Immunology SC Immunology GA 285VE UT WOS:000085408300036 PM 10671223 ER PT J AU Dimmock, JR Vashishtha, SC Stables, JP AF Dimmock, JR Vashishtha, SC Stables, JP TI Anticonvulsant properties of various acetylhydrazones, oxamoylhydrazones and semicarbazones derived from aromatic and unsaturated carbonyl compounds SO EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article DE semicarbazones; acetylhydrazones; oxamoylhydrazones; anticonvulsant; atomic charges ID ARYL SEMICARBAZONES; THIOSEMICARBAZONES AB Various acetylhydrazones, oxamoylhydrazones and semicarbazones were prepared as candidate anticonvulsants with a view to examining the viability of a putative binding site hypothesis. Atomic charge calculations were undertaken to determine the hydrogen bonding capacities of various molecules. The biological results obtained revealed that in general the acetylhydrazones and semicarbazones afforded good protection against convulsions while the oxamoylhydrazones were significantly less active. These data suggest that terminal electron-donating groups enhanced the hydrogen bonding capabilities and anticonvulsant properties of these molecules. (C) 2000 Editions scientifiques et medicales Elsevier SAS. C1 Univ Saskatchewan, Coll Pharm & Nutr, Saskatoon, SK S7N 5C9, Canada. NINDS, NIH, Bethesda, MD 20892 USA. RP Dimmock, JR (reprint author), Univ Saskatchewan, Coll Pharm & Nutr, Saskatoon, SK S7N 5C9, Canada. EM dimmock@skyway.usask.ca NR 22 TC 158 Z9 166 U1 1 U2 7 PU ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER PI PARIS PA 23 RUE LINOIS, 75724 PARIS, FRANCE SN 0223-5234 J9 EUR J MED CHEM JI Eur. J. Med. Chem. PD FEB PY 2000 VL 35 IS 2 BP 241 EP 248 DI 10.1016/S0223-5234(00)00123-9 PG 8 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 299WX UT WOS:000086221800006 PM 10758285 ER PT J AU Mikaels, A Livet, J Westphal, H de Lapeyriere, O Ernfors, P AF Mikaels, A Livet, J Westphal, H de Lapeyriere, O Ernfors, P TI A dynamic regulation of GDNF-family receptors correlates with a specific trophic dependency of cranial motor neuron subpopulations during development SO EUROPEAN JOURNAL OF NEUROSCIENCE LA English DT Article DE facial; gfr alpha; ret; trigeminal nucleus; GDNF ID C-RET PROTOONCOGENE; MICE LACKING GDNF; NEUROTROPHIC FACTOR; SENSORY NEURONS; CELL-DEATH; NERVOUS-SYSTEM; CUTANEOUS MECHANORECEPTORS; REQUIRE NEUROTROPHIN-3; GENE-EXPRESSION; TYROSINE KINASE AB Glial cell line-derived neurotrophic factor (GDNF) family ligands promote the survival of developing motor neurons in vivo and in vitro. However, not all neurons survive with any single ligand in culture and GDNF null mutant mice display only a partial motor neuron loss. An interesting possibility is that subpopulations of motor neurons based on their function and/or their myotopic organization require distinct members of GDNF family ligands. Because responsiveness to the different ligands depends on the expression of their cognate ligand-binding receptor we have herein addressed this issue by examining the expression of GDNF-family receptors (gfr) during development and in the adult in cranial motor nuclei subpopulations. We have furthermore examined the in vivo role of GDNF for cranial motor neuron subpopulations. The shared ret receptor was expressed in all somatic, branchial and visceral cranial embryonic motor nuclei examined, showing that they are all competent to respond to GDNF family ligands during development. At early stages of development both the GDNF receptor, gfr alpha 1, and the neurturin (NTN) receptor, gfr alpha 2, were expressed in the oculomotor, facial and spinal accessory, and only gfr alpha 1 in the trochlear, superior salivatory, trigeminal, hypoglossal and weakly in the dorsal motor nucleus of the vagus and the ambiguus nucleus. The abducens nucleus was negative for both gfr alpha 1 and gfr alpha 2. The artemin (ART) receptor, gfr alpha 3, was expressed only in the superior salivatory nucleus. A motor neuron subnuclei-specific expression of gfr alpha 1 and gfr alpha 2 was seen in the facial and trigeminal nuclei which corresponded to their dependence on GDNF in null mutant mice. We found that the expression was dynamic in these nuclei, which may reflect developmental changes in their trophic factor dependency. Analysis of GDNF null mutant mice revealed that the dynamic receptor expression is regulated by the ligand in vivo, indicating that the acquirement of changes in dependency could be ligand induced. Our results indicate that specific GDNF family ligands support selective muscle-motor neuron circuits during development. C1 Karolinska Inst, MBB, Mol Neurobiol Lab, S-17177 Stockholm, Sweden. Univ Mediterranee, INSERM, CNRS, IBDM,U382, F-13288 Marseille, France. NIH, Lab Mammalial Genes & Dev, Bethesda, MD 20892 USA. RP Ernfors, P (reprint author), Karolinska Inst, MBB, Mol Neurobiol Lab, S-17177 Stockholm, Sweden. RI livet, jean/C-2766-2012 NR 68 TC 28 Z9 28 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0953-816X J9 EUR J NEUROSCI JI Eur. J. Neurosci. PD FEB PY 2000 VL 12 IS 2 BP 446 EP 456 DI 10.1046/j.1460-9568.2000.00924.x PG 11 WC Neurosciences SC Neurosciences & Neurology GA 294UC UT WOS:000085927900004 PM 10712625 ER PT J AU Vicario-Abejon, C Collin, C Tsoulfas, P McKay, RDG AF Vicario-Abejon, C Collin, C Tsoulfas, P McKay, RDG TI Hippocampal stem cells differentiate into excitatory and inhibitory neurons SO EUROPEAN JOURNAL OF NEUROSCIENCE LA English DT Article DE BDNF; GABAergic; glutamatergic; NT-3; rat ID FIBROBLAST GROWTH-FACTOR; CENTRAL-NERVOUS-SYSTEM; NEUROTROPHIC FACTOR; PROGENITOR CELLS; IN-VITRO; PRECURSOR CELLS; MOUSE-BRAIN; CNS; TRANSPLANTATION; SURVIVAL AB Stem cell technology promises new and rapid advances in cell therapy and drug discovery. Clearly, the value of this approach will be limited by the differentiated functions displayed by the progeny of stem cells. The foetal and adult central nervous system (CNS) harbour stem cells that can be expanded in vitro and differentiate into immature neurons and glia. Surprisingly, we do not know if neurons derived from stem cells form synapses, a definitive feature of neuronal function. Neuronal differentiation is a complex process and in this paper we establish conditions that permit extensive maturation of neurons in the presence of neurotrophins. These conditions permit the differentiation of rat hippocampal stem cells into both excitatory (glutamatergic) and inhibitory (GABAergic) neurons. The proportion of excitatory and inhibitory synapses was strongly influenced by specific neurotrophins, and these responses reflect the region of origin of the stem cells in the brain. These data show that stem cells can be used to study mechanisms of excitation and inhibition in the nervous system. C1 NINDS, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. Univ Miami, Sch Med, Dept Neurol Surg, Miami, FL 33136 USA. Univ Miami, Sch Med, Miami Project, Miami, FL 33136 USA. RP Vicario-Abejon, C (reprint author), CSIC, Ctr Invest Biol, Velazquez 144, E-28006 Madrid, Spain. OI Tsoulfas, Pantelis/0000-0003-1974-6366 NR 58 TC 73 Z9 85 U1 0 U2 2 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0953-816X J9 EUR J NEUROSCI JI Eur. J. Neurosci. PD FEB PY 2000 VL 12 IS 2 BP 677 EP 688 DI 10.1046/j.1460-9568.2000.00953.x PG 12 WC Neurosciences SC Neurosciences & Neurology GA 294UC UT WOS:000085927900028 PM 10712648 ER PT J AU Panlilio, LV Weiss, SJ Schindler, CW AF Panlilio, LV Weiss, SJ Schindler, CW TI Stimulus compounding enhances conditioned suppression produced by cocaine-paired stimuli SO EXPERIMENTAL AND CLINICAL PSYCHOPHARMACOLOGY LA English DT Article ID APPETITIVE-AVERSIVE INTERACTIONS; SELF-ADMINISTERED COCAINE; BEHAVIORAL SENSITIZATION; DISCRIMINATIVE STIMULI; LATENT INHIBITION; OPERANT; TOLERANCE; RATS; AMPHETAMINE; REINFORCEMENT AB When 2 stimuli that occasion cocaine self-administration are presented in compound, their ability to increase cocaine-reinforced operant responding is substantially enhanced. The goal of the present experiment was to determine whether stimulus compounding could produce analogous enhancements of a classically conditioned drug effect. Food-maintained responding in rats was suppressed by a tone and a light that were individually paired with response-independent cocaine (3 mg/kg iv). This conditioned suppression was significantly enhanced when the stimuli were presented together in a stimulus-compounding test. The magnitude of this enhancement was similar to that in previous studies in which responding was suppressed by shock-paired stimuli. These results demonstrate that multiple drug-related cues interact in a predictable manner to influence both operant and classically conditioned behavior. C1 Natl Inst Drug Abuse, Preclin Pharmacol Sect, Behav Neurosci Branch, Intramural Res Program, Baltimore, MD 21224 USA. American Univ, Dept Psychol, Washington, DC 20016 USA. RP Panlilio, LV (reprint author), Natl Inst Drug Abuse, Preclin Pharmacol Sect, Behav Neurosci Branch, Intramural Res Program, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. FU NIDA NIH HHS [DA-08651-01A1] NR 77 TC 6 Z9 7 U1 1 U2 1 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 1064-1297 J9 EXP CLIN PSYCHOPHARM JI Exp. Clin. Psychopharmacol. PD FEB PY 2000 VL 8 IS 1 BP 6 EP 13 PG 8 WC Psychology, Biological; Psychology, Clinical; Pharmacology & Pharmacy; Psychiatry SC Psychology; Pharmacology & Pharmacy; Psychiatry GA 293GQ UT WOS:000085844000002 PM 10743900 ER PT J AU Taylor, RC Harris, NA Singleton, EG Moolchan, ET Heishman, SJ AF Taylor, RC Harris, NA Singleton, EG Moolchan, ET Heishman, SJ TI Tobacco craving: Intensity-related effects of imagery scripts in drug abusers SO EXPERIMENTAL AND CLINICAL PSYCHOPHARMACOLOGY LA English DT Article; Proceedings Paper CT 106th Annual Convention of the American-Psychological-Association CY AUG 15-16, 1998 CL SAN FRANCISCO, CALIFORNIA SP Amer Psychol Assoc ID SMOKING WITHDRAWAL SYMPTOMS; CUE REACTIVITY; URGES; COCAINE; SMOKERS; ALCOHOL; REINSTATEMENT; QUESTIONNAIRE; MANIPULATION; ABSTINENCE AB Two experiments were conducted to determine whether active imagery would elicit tobacco craving in smokers with histories of drug abuse who were not interested in quitting smoking. In Experiment 1, the authors used scripts that contained positive, negative, or neutral affective content with and without descriptions of smoking urge. Scripts with urge content and negative affect scripts increased subjective reports of tobacco craving. An interaction between affective manipulation and urge content was observed on self-reported mood. In Experiment 2, positive affect scripts that varied in amount of urge content produced an orderly increase in tobacco craving as a function of urge intensity, suggesting that changes were specific to the imagery manipulation. In both experiments, increases in tobacco craving were positively correlated with craving for drug of choice, suggesting that stimuli that engender smoking urges may occasion craving for other drugs of abuse. C1 Natl Inst Drug Abuse, Clin Pharmacol & Therapeut Branch, Baltimore, MD 21224 USA. Johns Hopkins Univ, Behav Therapy Res Ctr, Sch Med, Dept Psychiat & Behav Sci, Baltimore, MD 21218 USA. RP Heishman, SJ (reprint author), Natl Inst Drug Abuse, Clin Pharmacol & Therapeut Branch, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. OI Singleton, Edward G./0000-0003-3442-877X NR 50 TC 47 Z9 47 U1 4 U2 8 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 1064-1297 J9 EXP CLIN PSYCHOPHARM JI Exp. Clin. Psychopharmacol. PD FEB PY 2000 VL 8 IS 1 BP 75 EP 87 DI 10.1037/1064-1297.8.1.75 PG 13 WC Psychology, Biological; Psychology, Clinical; Pharmacology & Pharmacy; Psychiatry SC Psychology; Pharmacology & Pharmacy; Psychiatry GA 293GQ UT WOS:000085844000009 PM 10743907 ER PT J AU Hansson, SR Hoffman, BJ AF Hansson, SR Hoffman, BJ TI Transient expression of a functional serotonin transporter in Merkel cells during late gestation and early postnatal rat development SO EXPERIMENTAL BRAIN RESEARCH LA English DT Article DE fluoxetine; quinacrine; trigeminal sensory system; primary cell culture; whiskers ID LECTIN-BINDING ANALYSIS; NEURAL CREST CELLS; TRIGEMINAL SYSTEM; COMPREHENSIVE IMMUNOFLUORESCENCE; CUTANEOUS MECHANORECEPTORS; QUINACRINE FLUORESCENCE; SOMATOSENSORY CORTEX; NERVOUS-SYSTEM; MYSTACIAL PAD; GROWTH-FACTOR AB We and others have previously identified serotonin transporter mRNA throughout the trigeminal system in the whisker region, trigeminal ganglion, trigeminal nucleus and thalamic relay stations. In order to further implicate a role for the serotonin transporter in this sensory system, we have now characterized serotonin transporter gene expression and function in primary cultures from the rat snout, at several stages of gestation. In this study, we have demonstrated a transient expression of serotonin transporter mRNA in quinacrine-positive Merkel cells between embryonic day 16 and postnatal day 5. Peak levels of mRNA occurred at embryonic day 20 and postnatal day 1. Merkel cells in culture exhibited a transient, antidepressant-sensitive [H-3]-serotonin uptake, which was maximal at a time in culture corresponding to embryonic day 22 (day of birth). This transient uptake of serotonin suggests a role for this monoamine during a critical time period of the developing trigeminal sensory system. Regulation of extracellular serotonin levels by transporter activity may reflect the specific formation of the merkel cell-sensory neuron complex in an analogous mechanism by which serotonin modulates synaptogenesis in the central nervous system. C1 NIMH, Unit Mol Pharmacol, Lab Cellular & Mol Regulat, Bethesda, MD 20892 USA. Univ Lund, Wallenberg Neurosci Ctr, Dept Physiol & Neurosci, Div Mol Neurobiol, S-22362 Lund, Sweden. RP Hansson, SR (reprint author), NIMH, Unit Mol Pharmacol, Lab Cellular & Mol Regulat, Bethesda, MD 20892 USA. NR 54 TC 6 Z9 6 U1 0 U2 0 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0014-4819 J9 EXP BRAIN RES JI Exp. Brain Res. PD FEB PY 2000 VL 130 IS 3 BP 401 EP 409 DI 10.1007/s002219900242 PG 9 WC Neurosciences SC Neurosciences & Neurology GA 277RB UT WOS:000084946400011 PM 10706438 ER PT J AU Zhang, XP Morham, SG Langenbach, R Baggs, RB Young, DA AF Zhang, XP Morham, SG Langenbach, R Baggs, RB Young, DA TI Lack of cyclooxygenase-2 inhibits growth of teratocarcinomas in mice SO EXPERIMENTAL CELL RESEARCH LA English DT Article DE prostaglandin G/H synthase; cyclooxygenase-2; teratocarcinoma; embryonic stem cells; prostaglandin E2 ID PROSTAGLANDIN-G/H SYNTHASE; COLON-CANCER CELLS; EPITHELIAL-CELLS; ARACHIDONIC-ACID; GENE DISRUPTION; PHORBOL ESTER; MESSENGER-RNA; EXPRESSION; FIBROBLASTS; INDUCTION AB Two isoforms of cyclooxygenase (COX-1 or COX-2) have been identified in the prostanoid biosynthetic pathway. The constitutive form, COX-1, is thought to maintain cellular homeostasis and the inducible form, COX-2, is recognized as a primary response gene thought to be involved in modulating cell proliferation and differentiation. To further characterize the role of the cyclooxygenases in cell proliferation, differentiation, and tumorigenicity we developed embryonic stem (ES) cell lines which contain homozygous disruptions in either the COX-1 or the COX-2 gene. These lines were then examined in terms of their viability, proliferation, and in vitro differentiation potential. Our results demonstrate that the wild-type ES cells do not express either COX-1 or COX-2 until the cells undergo differentiation. And the lack of either cyclooxygenase has no apparent effect on ES cell proliferation in vitro, However, the absence of a functional COX-2 gene leads to a dramatic reduction in the formation and growth of teratocarcinomas that appear when ES cells are injected into syngeneic mice. Histological microscopy shows that the few very small tumors that were generated from ES cells lacking COX-2 appear more differentiated than tumors emerging from COX-1 -/- or wild-type cells by exhibiting greater keratinization in the areas of squamous epithelium and the ossification of bone-forming cartilage. We conclude that the presence of a functional COX-2 enzyme is necessary for the efficient growth of these teratocarcinomas in animals. (C) 2000 Academic Press. C1 Univ Rochester, Sch Med & Dent, Dept Med, Div Endocrinol & Metab,E Henry Keutmann Lab, Rochester, NY 14642 USA. Univ Rochester, Sch Med & Dent, Div Lab Anim Med, Rochester, NY 14642 USA. Myriad Genet Inc, Salt Lake City, UT 84108 USA. Natl Inst Environm Hlth Sci, Lab Expt Carcinogenesis & Mutagenesis, Res Triangle Pk, NC 27709 USA. RP Young, DA (reprint author), Univ Rochester, Sch Med & Dent, Dept Med, Div Endocrinol & Metab,E Henry Keutmann Lab, 601 Elmwood Ave, Rochester, NY 14642 USA. NR 33 TC 18 Z9 18 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0014-4827 J9 EXP CELL RES JI Exp. Cell Res. PD FEB 1 PY 2000 VL 254 IS 2 BP 232 EP 240 DI 10.1006/excr.1999.4758 PG 9 WC Oncology; Cell Biology SC Oncology; Cell Biology GA 280YP UT WOS:000085131600005 PM 10640421 ER PT J AU Sun, JK Iwata, T Zigler, JS Carper, DA AF Sun, JK Iwata, T Zigler, JS Carper, DA TI Differential gene expression in male and female rat lenses undergoing cataract induction by transforming growth factor-beta (TGF-beta) SO EXPERIMENTAL EYE RESEARCH LA English DT Article DE cataract; differential display; gamma B-crystallin; sex differences; lens; proteasome Z subunit; TGF-beta ID ANTERIOR SUBCAPSULAR CATARACTS; MESSENGER-RNA; GAMMA-CRYSTALLINS; INTERFERON-GAMMA; EYE LENS; DISPLAY; CELL; IDENTIFICATION; SUPERFAMILY; PROTEASOMES AB Epidemiologic studies in humans as well as immunohistologic studies in animals have demonstrated significant sex differences in the propensity to develop cataract. Several studies suggest that estrogen may play a protective role against cataractogenesis, Indeed, male and ovariectomized female rat lenses have a greater susceptibility to cataract induced by transforming growth factor-beta (TGF-beta) than do normal female lenses, However, in spite of the current evidence that estrogen may play a pivotal role in cataractogenesis, the molecular mechanisms behind this phenomenon are largely undetermined. Our study utilized the differential display procedure to examine gene up- and down-regulation in male, normal female and ovariectomized female rat lenses exposed to TGF-beta. Male and normal female rat lenses were cultured with or without 0.15 ng ml(-1) TGF-beta. Lenses were then harvested, and total RNA was isolated for analysis by reverse-transcriptase differential display. Differentially expressed mRNAs were subcloned, sequenced and identified through GenBank database searches, The original experiment was repeated with the addition of ovariectomized female TGF-beta (+/-) conditions, and all differential patterns of gene expression were verified using Northern blot and RT-PCR analysis. Screening of approximately 12% of the mRNA population led to the identification of 27 differentially expressed cDNAs. Notably, strong gender differences were found in expression levels of gamma B-crystallin. In addition, proteasome Z subunit was up-regulated in TGF-beta-treated male and ovariectomized female lenses, but was down-regulated in TGF-beta-treated normal female lenses, This pattern of expression is consistent with the increased susceptibility of male and ovariectomized lenses to TGF-beta-induced cataract. We conclude that differential display is a useful and expedient method for analysing changes in gene expression in the lens. Structural and functional studies of the genes identified in this study may further elucidate mechanisms underlying the TGF-beta-induced cataract formation and differential rates of cataractogenesis in males vs females. In particular, our data suggest that the role of proteasome Z subunit in TGF-beta-induced anterior subcapsular cataract warrants further investigation. (C) 2000 Academic Press. C1 NEI, NIH, Bethesda, MD 20892 USA. RP Carper, DA (reprint author), NEI, NIH, 9000 Rockville Pike,Bldg 6,Room 232, Bethesda, MD 20892 USA. NR 40 TC 12 Z9 14 U1 0 U2 0 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0014-4835 J9 EXP EYE RES JI Exp. Eye Res. PD FEB PY 2000 VL 70 IS 2 BP 169 EP 181 DI 10.1006/exer.1999.0771 PG 13 WC Ophthalmology SC Ophthalmology GA 284UZ UT WOS:000085352400005 PM 10655142 ER PT J AU Akin, C Kirshenbaum, AS Semere, T Worobec, AS Scott, LM Metcalfe, DD AF Akin, C Kirshenbaum, AS Semere, T Worobec, AS Scott, LM Metcalfe, DD TI Analysis of the surface expression of c-kit and occurrence of the c-kit Asp816Val activating mutation in T cells, B cells, and myelomonocytic cells in patients with mastocytosis SO EXPERIMENTAL HEMATOLOGY LA English DT Article DE mastocytosis; c-kit; mutation; hematopoiesis ID RECEPTOR TYROSINE KINASE; BLOOD MONONUCLEAR-CELLS; MURINE FETAL LIVER; PERIPHERAL-BLOOD; INDEPENDENT GROWTH; PROGENITOR CELLS; MAST-CELLS; STEM-CELLS; DIFFERENTIATION; LINE AB Objective. The Asp816Val c-kit activating mutation is detectable in the peripheral blood cells of some patients with mastocytosis and in lesional skin biopsies obtained from adult patients with urticaria pigmentosa, These observations led to the conclusion that this mutation is present in mast cells and mast cell precursors that express c-kit, However. the distribution of the Asp816Val mutation among hematopoietic lineages is unknown, To determine the distribution of the AspS16Val mutation among hematopoietic lineages and to explore its relationship to clinical disease, we examined cells bearing differentiation markers for myelomorlocytic cells as well as T and B lymphocytes, in both peripheral blood and bone marrow obtained from patients with mastocytosis. Materials and Methods. The presence of Asp816Val c-kit mutation in cells magnetically sorted from peripheral blood or bone marrow according to surface differentiation markers was studied by reverse transcriptase polymerase chain reaction (RT-PCR) restriction fragment length polymorphism (RFLP) analysis, The surface expression of c-kit was determined by flow cytometry. Results. The mutation was detectable by RT-PCR in at Least one cell lineage in the bone marrow in 7 of 7 patients examined and in the peripheral blood of 11 of 11 adult patients with urticaria pigmentosa and indolent disease. The mutation was identified most frequently in B cell?, and myeloid cells. Flow cytometric analysis demonstrated that the differentiated cells expressing mutated c-kit were negative for surface KIT. Conclusion. These results are consistent with the conclusion that the c-kit Asp816Val mutation occurs in an early progenitor cell and is carried by myelomonocytic cells, T cells, and B cells in addition to mast cells. However, unlike mast cells, these myelomonocgtic cells, T cells, and B cells in cells do not concomitantly express surface c-kit and thus may be less susceptible to the effects of this mutation. (C) 2000 International Society for Experimental Hematology. Published by Elsevier Science Inc. C1 NIAID, Lab Allerg Dis, NIH, Bethesda, MD 20892 USA. RP Akin, C (reprint author), NIAID, Lab Allerg Dis, NIH, Bldg 10,Room 11C205,10 Ctr Dr,MSC 1881, Bethesda, MD 20892 USA. NR 31 TC 132 Z9 136 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0301-472X J9 EXP HEMATOL JI Exp. Hematol. PD FEB PY 2000 VL 28 IS 2 BP 140 EP 147 DI 10.1016/S0301-472X(99)00145-9 PG 8 WC Hematology; Medicine, Research & Experimental SC Hematology; Research & Experimental Medicine GA 289GG UT WOS:000085611700003 PM 10706069 ER PT J AU Glazner, GW Mattson, MP AF Glazner, GW Mattson, MP TI Differential effects of BDNF, ADNF9, and TNF alpha on levels of NMDA receptor subunits, calcium homeostasis, and neuronal vulnerability to excitotoxicity SO EXPERIMENTAL NEUROLOGY LA English DT Article DE BDNF; TNF alpha; ADNF; AMPA; excitotoxicity; NF-kappa B; glutamate ID CULTURED HIPPOCAMPAL-NEURONS; CEREBELLAR GRANULE CELLS; GLUTAMATE-INDUCED NEUROTOXICITY; NECROSIS-FACTOR-ALPHA; RAT CORTICAL-NEURONS; NERVE GROWTH-FACTOR; PROTEIN-KINASE-C; NEUROTROPHIC FACTOR; SYNAPTIC TRANSMISSION; CEREBRAL-ISCHEMIA AB Calcium influx through N-methyl-D-aspartate (NMDA) receptors can result in neuronal apoptosis or necrosis and may play a pivotal role in neuronal death in many different neurodegenerative diseases. In the present study we employed primary neuronal cultures and three different excitoprotective factors, brain-derived neurotrophic factor (BDNF), activity-dependent neurotrophic factor (ADNF9), and tumor necrosis factor alpha (TNF alpha), to elucidate the mechanisms whereby trophic factors modify the excitotoxic process. Neurons pretreated with BDNF exhibited increased levels of the NMDAreceptor subunits NR1 and NR2A which was associated with increased calcium responses to NMDA and vulnerability to excitotoxic necrosis and reduced vulnerability to apoptosis. ADNF9 and TNF alpha suppressed calcium responses to glutamate and protected neurons against both excitotoxic necrosis and apoptosis, but had no effect on levels of NMDA receptor subunits. Inhibition of phosphorylation and DNA binding of NF-kappa B, by H7 and KB decoy DNA, respectively; suggest that the extitotoxic-modulating actions of BDNF are mediated by kinases, while those of ADNF9 and TMF alpha are mediated by both kinases and the transcription factor NF-kappa B. Our data show that, whereas BDNF increases neuronal responses to glutamate while ADNF9 and TNF alpha decrease the same, all three protect against excitotoxic apoptosis. (C) 2000 Academic Press. C1 Univ Kentucky, Sanders Brown Ctr Aging, Lexington, KY 40536 USA. Univ Kentucky, Dept Anat & Neurobiol, Lexington, KY 40536 USA. RP Mattson, MP (reprint author), NIA, Neurosci Lab, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012 FU NINDS NIH HHS [NS29001, NS35253]; NINR NIH HHS [NRSA467305] NR 64 TC 72 Z9 74 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0014-4886 J9 EXP NEUROL JI Exp. Neurol. PD FEB PY 2000 VL 161 IS 2 BP 442 EP 452 DI 10.1006/exnr.1999.7242 PG 11 WC Neurosciences SC Neurosciences & Neurology GA 290FW UT WOS:000085666500002 PM 10686066 ER PT J AU Conejero-Goldberg, C Tornatore, C Abi-Saab, W Monaco, MC Dillon-Carter, O Vawter, M Elsworth, J Freed, W AF Conejero-Goldberg, C Tornatore, C Abi-Saab, W Monaco, MC Dillon-Carter, O Vawter, M Elsworth, J Freed, W TI Transduction of human GAD67 cDNA into immortalized striatal cell lines using an Epstein-Barr virus-based plasmid vector increases GABA content SO EXPERIMENTAL NEUROLOGY LA English DT Article DE GAD; GABA; EBV; immortalized cell lines; SV40 large T antigen ID LARGE T-ANTIGEN; GLUTAMIC-ACID DECARBOXYLASE; CEREBROSPINAL-FLUID; PARKINSONS-DISEASE; EXPRESSION VECTOR; MAMMALIAN-CELLS; GENE DELIVERY; NEURONS; RELEASE; RAT AB The M213-20 and M213-1L cell lines were immortalized from rat striatum using the tsA58 allele of the SV40 large T antigen, contain the GAD enzyme, and produce GABA (Giordano ct al., 1994, Exp. Neurol 124:395-400). Cell lines that produce large amounts of GABA may be useful for transplantation into the brain in conditions such as Huntington's disease or epilepsy, where localized application of GABA may be of therapeutic value. We have explored the potential use of the pREP10 plasmid vector, which replicates episomally, to increase GAD expression and GABA production in M213-20 and M213-1L cells. Human GAD(67) cDNA was transfected into M213-20 and M213-1L, and subclones were isolated with hygromycin selection. Immunochemical studies showed increased GAD(67) expression compared to the parent M213-20 and M213-1L cell lines. Staining for the EBNA antigen and Southern blots demonstrated that the pREP10 plasmid was stably maintained in the cells for at least 12-15 months in culture. Several clones were isolated in which GABA concentrations were increased by as much as 4-fold (M213-1L) or 44-fold (M213-20) compared to the parent cell lines or 12-fold (M213-1L) and 94-fold (M213-20) greater than rat striatal tissue (1.678 +/- 0.4 mu mol/g prot). The ability of these cells to continue to produce large amounts of GABA while being maintained in culture for extended periods suggests that similar methods might be used with human cell lines to produce cells that can be transplanted into the brain to deliver GABA for therapeutic purposes. (C) 2000 Academic Press. C1 NIDA, Cellular Neurobiol Branch, Dev & Plast Sect, Baltimore, MD 21224 USA. NINDS, Lab Mol Med & Neurosci, Bethesda, MD 20892 USA. Yale Univ, Dept Psychiat, New Haven, CT 06519 USA. Yale Univ, Dept Neuropsychopharmacol, New Haven, CT 06519 USA. RP NIDA, Cellular Neurobiol Branch, Dev & Plast Sect, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 46 TC 19 Z9 20 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 0014-4886 EI 1090-2430 J9 EXP NEUROL JI Exp. Neurol. PD FEB PY 2000 VL 161 IS 2 BP 453 EP 461 DI 10.1006/exnr.1999.7258 PG 9 WC Neurosciences SC Neurosciences & Neurology GA 290FW UT WOS:000085666500003 PM 10686067 ER PT J AU Messam, CA Hou, J Major, EO AF Messam, CA Hou, J Major, EO TI Coexpression of nestin in neural and glial cells in the developing human CNS defined by a human-specific anti-nestin antibody SO EXPERIMENTAL NEUROLOGY LA English DT Article DE stem and progenitor cells; intermediate filament; glia; human CNS ID INTERMEDIATE FILAMENT NESTIN; CENTRAL-NERVOUS-SYSTEM; ADULT-MOUSE BRAIN; STEM-CELLS; PRECURSOR CELLS; SUBEPENDYMAL CELLS; GROWTH-FACTOR; SPINAL-CORD; EXPRESSION; ASTROCYTES AB The presence of the intermediate filament protein nestin has been the predominant marker used to describe stem and progenitor cells in the mammalian CNS. In this study, a 998-bp fragment in the 3' region of the nestin mRNA was cloned from human fetal brain cells (HFBC). The nucleotide sequence of the cloned cDNA revealed 21 differences with the previously published human nestin sequence, resulting in 17 amino acid changes. A 150 amino-acid fragment derived from the cloned nestin cDNA was coupled to glutathione S-transferase and used as an immunogen to generate a rabbit polyclonal antiserum that selectively detects human nestin, HFBC that proliferated in response to basic fibroblast growth factor incorporated 5-bromo-2'-deoxyuridine into their nuclei and immunostained for nestin, indicating nestin expression in proliferating CNS progenitor cells. In all cell cultures, nestin contained with the neuroepithelial cell. marker vimentin. A small subset of nestin-stained cells (1-2%) immunostained with neuronal marker MAP-2 during the first week and after 4 weeks in culture. However, during the first week in culture, approximately 10-30% of the total cell population of HFBC stained for the glial cell marker GFAP, and nearly all coimmunostained for nestin, After 4 weeks in culture, a subset of GFAP-positive cells emerged that no longer contained with nestin, These results describe nestin expression not only in CNS progenitor cells but also in the cells which were in transition from a progenitor stage to glial differentiation. Collectively, these data suggest a differential temporal regulation of nestin expression during glial and neuronal cell differentiation. (C) 2000 Academic Press. C1 NINDS, NIH, Lab Mol Med & Neurosci, Bethesda, MD 20892 USA. RP NINDS, NIH, Lab Mol Med & Neurosci, Bethesda, MD 20892 USA. EM messamc@ninds.nih.gov NR 45 TC 142 Z9 153 U1 0 U2 6 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0014-4886 EI 1090-2430 J9 EXP NEUROL JI Exp. Neurol. PD FEB PY 2000 VL 161 IS 2 BP 585 EP 596 DI 10.1006/exnr.1999.7319 PG 12 WC Neurosciences SC Neurosciences & Neurology GA 290FW UT WOS:000085666500014 PM 10686078 ER PT J AU McAreavey, D Exner, DV Curtin, EL Domanski, MJ AF McAreavey, D Exner, DV Curtin, EL Domanski, MJ TI beta-blockers in heart failure: recently completed and ongoing clinical trials SO EXPERT OPINION ON INVESTIGATIONAL DRUGS LA English DT Article DE beta-blocker; clinical trial; heart failure; mortality; prognosis ID LEFT-VENTRICULAR DYSFUNCTION; IDIOPATHIC DILATED CARDIOMYOPATHY; DOUBLE-BLIND; MYOCARDIAL-INFARCTION; RANDOMIZED EVALUATION; CONGESTIVE CARDIOMYOPATHY; ADRENERGIC-BLOCKADE; CARVEDILOL; METOPROLOL; MORTALITY AB beta-blockers have emerged as an important therapy in patients with symptomatic left ventricular systolic dysfunction. Early studies demonstrated that beta-blocker therapy improved left ventricular function, reduced neurohumoral activity and reduced heart failure symptoms in these patients. While none of these small studies demonstrated a significant benefit in terms of overall survival, several meta-analyses suggested that beta-blocker therapy could, in fact, reduce mortality in patients with left ventricular systolic dysfunction and mild to moderate heart failure symptoms (New York Heart Association class II or III). Three large, recently completed, trials have confirmed the benefit of beta-blockade in these patients. This report reviews some of the initial clinical studies of beta-blockade in heart failure, examines the findings of the three large multicentre trials and other relevant research. Finally, ongoing trials designed to assess the relative efficacy of different beta-blockers and evaluate the utility of beta-blockade in specific subsets of patients with heart failure are discussed. C1 NHLBI, Clin Trials Res, Bethesda, MD 20892 USA. SmithKline Beecham Pharma, Clin Res, Cardiovasc Therapeut Unit, Collegeville, PA USA. RP McAreavey, D (reprint author), NHLBI, Clin Trials Res, 6701 Rockledge Dr,Room 8146, Bethesda, MD 20892 USA. NR 57 TC 1 Z9 1 U1 0 U2 0 PU ASHLEY PUBL LTD PI LONDON PA 1ST FL, THE LIBRARY, 1 SHEPHERDS HILL HIGHGATE, LONDON N6 5QJ, ENGLAND SN 1354-3784 J9 EXPERT OPIN INV DRUG JI Expert Opin. Investig. Drugs PD FEB PY 2000 VL 9 IS 2 BP 415 EP 428 DI 10.1517/13543784.9.2.415 PG 14 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 344VK UT WOS:000088780000016 PM 11060685 ER PT J AU Zhao, WQ Meiri, N Xu, H Cavallaro, S Quattrone, A Zhang, L Alkon, DL AF Zhao, WQ Meiri, N Xu, H Cavallaro, S Quattrone, A Zhang, L Alkon, DL TI Spatial learning induced changes in expression of the ryanodine type II receptor in the rat hippocampus SO FASEB JOURNAL LA English DT Article DE water maze task; learning and memory; calcium; mRNA; in situ hybridization ID CALCIUM RELEASE CHANNEL; LONG-TERM POTENTIATION; DENTATE GYRUS; MEMORY; CA2+; KINASE; BRAIN; IDENTIFICATION; LOCALIZATION; DEPRESSION AB Calcium signaling critical to neural functions is mediated through Ca2+ channels localized on both the plasma membrane and intracellular organelles such as endoplasmic reticulum, Whereas Ca2+ influx occurs via the voltage- or/and ligand-sensitive Ca2+ channels, Ca2+ release from intracellular stores that amplifies further the Ca2+ signal is thought to be involved in more profound and lasting changes in neurons, The ryanodine receptor, one of the two major intracellular Ca2+ channels, has been an important target for studying Ca2+ signaling in brain functions, including learning and memory, due to its characteristic Ca2+-induced Ca2+ release, In this study, we report regional and cellular distributions of the type-2 ryanodine receptor (RyR2) mRNA in the rat brain, and effects of spatial learning on RyR2 gene expression at mRNA and protein levels in the rat hippocampus, Using in situ hybridization, reverse transcription polymerase chain reaction, and ribonuclease protection assays, significant increases in RyR2 mRNA were found in the hippocampus of rats trained in an intensive water maze task, With immunoprecipitation and immunoblotting, protein levels of RyR2 were also demonstrated to be increased in the microsomal fractions prepared from hippocampi of trained rats, These results suggest that RyR2, and hence the RyR2-mediated Ca2+ signals, may be involved in memory processing after spatial learning. The increases in RyR2 mRNA and protein at 12 and 24 h after training could contribute to more permanent changes such as structural modifications during long-term memory storage. C1 NINDS, Lab Adapt Syst, NIH, Bethesda, MD 20892 USA. NIMH, Behav Endocrinol Branch, NIH, Bethesda, MD 20892 USA. Agr Res Org, Volcani Ctr, Inst Anim Sci, IL-50250 Bet Dagan, Israel. CNR, Inst Bioimaging & Pathophysiol Cent Nervous Syst, Catania, Italy. IRCCS, Oasai Inst Res Mental Retardat & Brain Aging, Troina, EN, Italy. RP Zhao, WQ (reprint author), NINDS, Lab Adapt Syst, NIH, Bldg 36,Room 4A22,36 Convent Dr, Bethesda, MD 20892 USA. RI Cavallaro, Sebastiano/F-3104-2010; OI Cavallaro, Sebastiano/0000-0001-7590-1792; Quattrone, Alessandro/0000-0003-3333-7630 NR 43 TC 58 Z9 61 U1 0 U2 1 PU FEDERATION AMER SOC EXP BIOL PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0892-6638 J9 FASEB J JI Faseb J. PD FEB PY 2000 VL 14 IS 2 BP 290 EP 300 PG 11 WC Biochemistry & Molecular Biology; Biology; Cell Biology SC Biochemistry & Molecular Biology; Life Sciences & Biomedicine - Other Topics; Cell Biology GA 281WQ UT WOS:000085184800008 PM 10657985 ER PT J AU Lindner, G Menrad, A Gherardi, E Merlino, G Welker, P Handjiski, B Roloff, B Paus, R AF Lindner, G Menrad, A Gherardi, E Merlino, G Welker, P Handjiski, B Roloff, B Paus, R TI Involvement of hepatocyte growth factor/scatter factor and Met receptor signaling in hair follicle morphogenesis and cycling SO FASEB JOURNAL LA English DT Article DE apoptosis; hair growth; HGF/SF; keratinocytes; murine hepatocyte growth factor receptor ID FACTOR SCATTER FACTOR; EPITHELIAL-CELLS; REGRESSION CATAGEN; MURINE SKIN; TYROSINE KINASE; CYCLOSPORINE-A; ORGAN-CULTURE; BETA-CATENIN; MOUSE SKIN; IN-VIVO AB HGF/SF and its receptor (Met) are principal mediators of mesenchymal-epithelial interactions in several different systems and have recently been implicated in the control of hair follicle (HF) growth, We have studied their expression patterns during HF morphogenesis and cycling in C57BL/6 mice, whereas functional hair growth effects of HGF/SF were assessed in vivo by analysis of transgenic mice and in skin organ culture. In normal mouse skin, follicular expression of HGF/SF and Met was strikingly localized: HGF/SF was found only in the HF mesenchyme (dermal papilla fibroblasts) and Met in the neighboring hair bulb keratinocytes, Both HGF/SF and Met expression peaked during the initial phases of HF morphogenesis, the stage of active hair growth (early and mid anagen), and during the apoptosis-driven HF regression (catagen), Met+ cells in the regressing epithelial strand appeared to be protected from undergoing apoptosis. Compared to wild-type controls, transgenic mice overexpressing HGF/SF under the control of the MT-I promoter had twice as many developing HF and displayed accelerated HF development on postnatal day 3, They also showed significant catagen retardation on P17, In organ culture and in vivo, HGF/SF i.c. resulted in a significant catagen retardation. These results demonstrate an important role of HGF/SF and Met in murine hair growth control and suggest that Met-mediated signaling might be exploited for therapeutic manipulation of human hair growth disorders.-Lindner, G., Menrad, A., Gherardi, E., Merlino, G,, Welker, P., Handjiski, B., Roloff, B,, Paus, R. Involvement of hepatocyte growth factor/scatter factor and Met receptor signaling in hair follicle morphogenesis and cycling. C1 Univ Hamburg, Hosp Eppendorf, Dept Dermatol, D-20246 Hamburg, Germany. Humboldt Univ, Dept Dermatol, D-1086 Berlin, Germany. Schering AG, Res Labs, D-1000 Berlin, Germany. MRC Ctr, Growth Factors Grp, Dept Oncol, Cambridge, England. NCI, Mol Biol Lab, Bethesda, MD 20892 USA. RP Paus, R (reprint author), Univ Hamburg, Hosp Eppendorf, Dept Dermatol, Martinistr 52, D-20246 Hamburg, Germany. NR 71 TC 86 Z9 89 U1 1 U2 5 PU FEDERATION AMER SOC EXP BIOL PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0892-6638 J9 FASEB J JI Faseb J. PD FEB PY 2000 VL 14 IS 2 BP 319 EP 332 PG 14 WC Biochemistry & Molecular Biology; Biology; Cell Biology SC Biochemistry & Molecular Biology; Life Sciences & Biomedicine - Other Topics; Cell Biology GA 281WQ UT WOS:000085184800011 PM 10657988 ER PT J AU Anson, RM Hudson, E Bohr, VA AF Anson, RM Hudson, E Bohr, VA TI Mitochondrial endogenous oxidative damage has been overestimated SO FASEB JOURNAL LA English DT Article DE 8-hydroxy-2 '-deoxyguanosine; 8-hydroxydeoxy-guanosine; 8-oxoguanine; 8-oxodG; aging; mitochondria ID FORMAMIDOPYRIMIDINE DNA GLYCOSYLASE; NUCLEAR-DNA; CELLS; REPAIR; DISEASE AB The oxidatively induced DNA lesion 8-oxo-dG in mitochondrial DNA (mtDNA) is commonly used as a marker for oxidative damage to mitochondria, which in turn is thought to be a fundamental cause of aging. For years, mitochondrial levels of 8-oxo-dG were believed to be similar to 10-fold higher in mtDNA than in nuclear DNA even in normal, young animals. However, studies in our own and other laboratories have shown that this lesion is efficiently repaired. Also, mutational consequences specific to 8-oxo-dG (G to T transversions) are rarely reported, In the present study, we showed that the levels of damage measured using high-pressure liquid chromatography/electrochemical detection and an enzymatic/Southern blot assay were comparable. The latter assay does not require isolation of mitochondria, and so this assay was then used to determine the level of in vivo damage present in rat Liver mtDNA both with and without organelle isolation. Levels of 8-oxo-dG are approximately threefold higher when measured in mtDNA purified from isolated mitochondria than when measured without prior mitochondrial isolation. Furthermore, most genomes were free of endogenous enzyme-sensitive sites (i.e., they did not contain 8-oxo-dG), and only after mitochondrial isolation were levels higher in mtDNA than in a nuclear sequence. C1 NIA, Mol Genet Lab, GRC, NIH, Baltimore, MD 21224 USA. RP Bohr, VA (reprint author), NIA, Cellular & Mol Biol Lab, NIH, Baltimore, MD 21224 USA. NR 25 TC 58 Z9 61 U1 0 U2 3 PU FEDERATION AMER SOC EXP BIOL PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0892-6638 J9 FASEB J JI Faseb J. PD FEB PY 2000 VL 14 IS 2 BP 355 EP 360 PG 6 WC Biochemistry & Molecular Biology; Biology; Cell Biology SC Biochemistry & Molecular Biology; Life Sciences & Biomedicine - Other Topics; Cell Biology GA 281WQ UT WOS:000085184800014 PM 10657991 ER PT J AU Dozortsev, D Coleman, A Nagy, P Diamond, MP Ermilov, A Weier, U Liyanage, M Reid, T AF Dozortsev, D Coleman, A Nagy, P Diamond, MP Ermilov, A Weier, U Liyanage, M Reid, T TI Nucleoli in a pronuclei-stage mouse embryo are represented by major satellite DNA of interconnecting chromosomes SO FERTILITY AND STERILITY LA English DT Article; Proceedings Paper CT XVI World Congress on Fertility and Sterility / 54th Annual Meeting of the American-Society-for-Reproductive-Medicine CY OCT 03-09, 1998 CL SAN FRANCISCO, CALIFORNIA SP Amer Soc Reprod Med, Int Federat Fertil & Steril DE zygote; nucleoli; alpha-satellite; centromeres; telomeres ID CHROMATIN; HYBRIDIZATION; ARRANGEMENTS; ASSOCIATION; INSITU; CELLS AB Objective: To investigate the arrangement of chromosomes within pronuclei-stage mouse zygotes. Design: In vitro study. Setting: Academic medical center. Patient(s): None. Intervention(s): None. Main Outcome Measure(s): Location of major alpha-satellite DNA, centromeres, and telomeres, and relative location of chromosomes. Result(s): Chromosomes appeared to be oriented inward by centromeres and to be interconnected by major alpha-satellite DNA, which appeared to be the sole DNA component of the nucleoli. This chromosomal arrangement persisted throughout interphase. Chromosomal painting failed to identify chromosomal ordering within pronuclei. Conclusion(s): Pronuclear nucleoli are represented by alpha-satellite sequences of interconnecting chromosomes that hold all chromosomes together during interphase. Chromosomes within the pronucleus are randomly positioned relative to each other. (Fertil Steril(R) 2000;73:366-71. (C)2000 by American Society for Reproductive Medicine.). C1 Wayne State Univ, Dept Obstet & Gynecol, Div Reprod Endocrinol, Detroit, MI USA. European Hosp, Rome, Italy. Univ Calif Berkeley, Lawrence Berkeley Lab, Berkeley, CA 94720 USA. NCI, Genet Lab, Div Basic Sci, Bethesda, MD 20892 USA. NIH, Genome Technol Branch, Natl Human Genome Res Inst, Bethesda, MD 20892 USA. RP Dozortsev, D (reprint author), Hutzel Hosp, Dept Obstet & Gynecol, 4707 St Antoine Blvd, Detroit, MI 48201 USA. OI Diamond, Michael/0000-0001-6353-4489 NR 19 TC 15 Z9 16 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0015-0282 J9 FERTIL STERIL JI Fertil. Steril. PD FEB PY 2000 VL 73 IS 2 BP 366 EP 371 DI 10.1016/S0015-0282(99)00491-4 PG 6 WC Obstetrics & Gynecology; Reproductive Biology SC Obstetrics & Gynecology; Reproductive Biology GA 281UF UT WOS:000085178600031 PM 10685545 ER PT J AU Bialkowski, K Kasprzak, KS AF Bialkowski, K Kasprzak, KS TI Activity of the antimutagenic enzyme 8-oxo-2 '-deoxyguanosine 5 '-triphosphate pyrophosphohydrolase (8-oxo-dGTPase) in cultured Chinese hamster ovary cells: Effects of cell cycle, proliferation rate, and population density SO FREE RADICAL BIOLOGY AND MEDICINE LA English DT Article DE 8-oxo-dGTPase; antimutagenic enzyme; free radicals; MTH1; MutT; nucleotide oxidative damage ID ESCHERICHIA-COLI; DNA-SYNTHESIS; NUCLEOSIDE TRIPHOSPHATASE; ENCODING 8-OXO-DGTPASE; TRANSVERSION MUTATION; REPLICATION FIDELITY; MUTAGENIC SUBSTRATE; OXIDATIVE STRESS; MESSENGER-RNA; MUTT PROTEIN AB Mammalian 8-oxo-2'-deoxyguanosine 5'-triphosphate pyrophosphohydrolases (8-oxo-dGTPases), such as MTH1, are believed to play the same antimutagenic role as their bacterial homologues, like MutT. Both decompose promutagenic 8-oxo-dGTP, a product of active oxygen's attack on dGTP. It is not known how 8-oxo-dGTPase expression and function are regulated. Therefore, we investigated the effect of cell population density, proliferation rate, and cell cycle phase on 8-oxo-dGTPase specific activity in cultured Chinese hamster ovary K1-BH4 (CHO) cells. With increasing cell population density (from 30 to 95% confluence), the activity of 8-oxo-dGTPase per milligram protein decreased by 33% (p = .007 by ANOVA) while cells shifted by 9% into the G(0)/G(1) phase, with a 5% drop in cells in S phase. Importantly, inhibition of the cells' proliferation rate by calf serum deprivation caused a more dramatic 23% shift toward the G(0)/G(1) phase and a 25% drop in S phase, but had no effect on 8-oxo-dGTPase activity. Likewise, no differences in the enzyme activity were observed within cell populations of different cell cycle phases separated by centrifugal elutriation. Thus, the present results exclude cell cycle-dependent regulation of 8-oxo-dGTPase activity in CHO cells or its simple dependence on proliferation rate. The observed decrease of 8-oxo-dGTPase activity with increasing cell population density might be related to augmentation of cell-to-cell contact. (C) 2000 Elsevier Science Inc. C1 L Rydygier Univ, Sch Med Sci, Dept Clin Biochem, PL-85092 Bydgoszcz, Poland. NCI, Frederick Canc Res & Dev Ctr, Comparat Carcinogenesis Lab, Frederick, MD USA. RP Bialkowski, K (reprint author), L Rydygier Univ, Sch Med Sci, Dept Clin Biochem, Ul Karlowicza 24, PL-85092 Bydgoszcz, Poland. RI Bialkowski, Karol/E-2328-2014 NR 33 TC 11 Z9 12 U1 0 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0891-5849 J9 FREE RADICAL BIO MED JI Free Radic. Biol. Med. PD FEB 1 PY 2000 VL 28 IS 3 BP 337 EP 344 DI 10.1016/S0891-5849(99)00246-4 PG 8 WC Biochemistry & Molecular Biology; Endocrinology & Metabolism SC Biochemistry & Molecular Biology; Endocrinology & Metabolism GA 289KD UT WOS:000085619000004 PM 10699744 ER PT J AU Manal, K McClay, I Stanhope, S Richards, J Galinat, B AF Manal, K McClay, I Stanhope, S Richards, J Galinat, B TI Comparison of surface mounted markers and attachment methods in estimating tibial rotations during walking: an in vivo study SO GAIT & POSTURE LA English DT Article DE surface-tracking marker; tibia; distal lateral shank; tibial rotation ID GAIT; KINEMATICS; KNEE; MOVEMENT; KINETICS; SYSTEM; MOTION AB The overall goal of this work was to determine an optimal surface-tracking marker set Ibr tracking motion of the tibia during natural cadence walking. Eleven different marker sets were valuated. The marker sets differed in location they were attached to the shank. the method used to attach the marker sets to the segment and the physical characteristics of the marker sets. Angular position during stance for each marker set was expressed relative to the orientation of the tibia as measured using bone anchored markers. A marker set consisting of four markers attached to a rigid shell positioned over the distal lateral shank and attached to the leg using an underwrap attachment yielded the brst estimate of tibial rotation. Rotational deviations of +/- 2 degrees about the medio-lateral and antero-posterior axes. and +/- 4 degrees about the longitudinal axis did occur even when using the optimal set of markers. (C) 2000 Elsevier Science B.V. All rights reserved. C1 Univ Delaware, Dept Mech Engn, Newark, DE 19716 USA. NIH, Biomech Lab, Bethesda, MD 20982 USA. Delaware Orthopaed Ctr, Newark, DE 19716 USA. RP Manal, K (reprint author), Univ Delaware, Dept Mech Engn, Newark, DE 19716 USA. NR 20 TC 142 Z9 144 U1 0 U2 13 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0966-6362 J9 GAIT POSTURE JI Gait Posture PD FEB PY 2000 VL 11 IS 1 BP 38 EP 45 DI 10.1016/S0966-6362(99)00042-9 PG 8 WC Neurosciences; Orthopedics; Sport Sciences SC Neurosciences & Neurology; Orthopedics; Sport Sciences GA 287ND UT WOS:000085511400005 PM 10664484 ER PT J AU Licht, T Goldenberg, SK Vieira, WD Gottesman, MM Pastan, I AF Licht, T Goldenberg, SK Vieira, WD Gottesman, MM Pastan, I TI Drug selection of MDR1-transduced hematopoietic cells ex vivo increases transgene expression and chemoresistance in reconstituted bone marrow in mice SO GENE THERAPY LA English DT Article DE gene therapy; bone marrow transplantation; chemotherapy; multidrug resistance; selectable marker genes ID MULTIDRUG-RESISTANCE GENE; P-GLYCOPROTEIN EXPRESSION; HUMAN MDR1 GENE; STEM-CELLS; PROGENITOR CELLS; RETROVIRAL TRANSFER; TRANSPLANTED MICE; MEDIATED TRANSFER; PERIPHERAL-BLOOD; PRECURSOR CELLS AB The MDR1 (multidrug resistance) gene, transferred to hematopoietic cells, is expected to protect them from anticancer chemotherapy and may serve as a selectable marker, restoring gene expression in vivo. Appropriate selection strategies, however, need to be established To investigate whether preselection ex vivo affects chemoresistance, murine bone marrow cells were retrovirally transduced with high-titer or, as a model for suboptimal gene:expression, low-titer retroviruses and exposed to daunomycin or colchicine for 48-96 h. Selection significantly increased chemoresistance of clonogenic progenitor cells. in tissue culture, the entire target population was rendered highly drug resistant after MDR1 transfer with high-titer viruses. if transduction was performed under suboptimal conditions, drug selection increased the frequency of chemoresistant colonies up to 40% over the number of unselected cells. Colchicine and daunomycin were equally efficient in increasing drug resistance ex vivo, but colchicine-preselected cells rescued lethally irradiated mice under conditions where daunomycin-selected bone marrow cells failed to do so. Hence, while hematopoietic cells can be protected by MDR1, the selection strategy is critical for repopulation of bone marrow with transduced cells. Preselection in culture before transplantation significantly increased P-gp expression and chemoresistance in vivo in mice reconstituted with transduced bone marrow cells. This study may help to facilitate the use of MDR1 as a selectable marker in gene therapy of the hematopoietic system. C1 NCI, Mol Biol Lab, Natl Canc Inst, Bethesda, MD 20892 USA. NCI, Cell Biol Lab, Natl Canc Inst, Bethesda, MD 20892 USA. RP Pastan, I (reprint author), NCI, Mol Biol Lab, Natl Canc Inst, Bldg 37,Room 4E16,37Convent Dr MSC 4255, Bethesda, MD 20892 USA. NR 52 TC 20 Z9 23 U1 0 U2 1 PU STOCKTON PRESS PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0969-7128 J9 GENE THER JI Gene Ther. PD FEB PY 2000 VL 7 IS 4 BP 348 EP 358 DI 10.1038/sj.gt.3301087 PG 11 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity; Research & Experimental Medicine GA 285FR UT WOS:000085379500012 PM 10694816 ER PT J AU Shakhov, AN Rubtsov, AV Lyakhov, IG Tumanov, AV Nedospasov, SA AF Shakhov, AN Rubtsov, AV Lyakhov, IG Tumanov, AV Nedospasov, SA TI SPLASH (PLA(2)IID), a novel member of phospholipase A(2) family, is associated with lymphotoxin deficiency SO GENES AND IMMUNITY LA English DT Article DE phospholipase A(2); subtractive cloning; lymphotoxin; knock-out mice; spleen; tumor suppressor genes ID TUMOR-NECROSIS-FACTOR; MULTIPLE INTESTINAL NEOPLASIA; SECONDARY LYMPHOID-TISSUES; LOW-MOLECULAR-WEIGHT; ABNORMAL-DEVELOPMENT; MAJOR MODIFIER; BETA-RECEPTOR; MOUSE; MICE; GENE AB Lymphotoxin (LT) deficient mice have profound defects in the splenic microarchitecture associated with defective expression on certain gene products, including chemokines. By using subtraction cloning of splenic cDNA from wild-type and LT alpha or TNF/LT alpha double deficient mice we isolated a novel murine gene encoding a secretory type phospholipase A(2), called SPLASH. The two major alternative transcripts of SPLASH gene are predominantly expressed in lymphoid tissues, such as spleen and lymph nodes. SPLASH maps to the distal part of chromosome 4, to which several cancer-related loci have been also mapped. C1 NCI, FCRDC, SAIC Frederick, Intramural Res Support Program, Frederick, MD 21702 USA. NCI, FCRDC, Div Basic Sci, Mol Immunoregulat Lab, Frederick, MD 21702 USA. ETH, Inst Toxicol, Schwerzenbach, Switzerland. Russian Acad Sci, Engelhardt Inst Mol Biol, Lab Mol Immunol, Moscow 117984, Russia. Moscow MV Lomonosov State Univ, Belozersky Inst Phys Chem Biol, Moscow 119899, Russia. RP Shakhov, AN (reprint author), NCI, FCRDC, SAIC Frederick, Intramural Res Support Program, Bldg 560,Rm 31-33, Frederick, MD 21702 USA. RI Nedospasov, Sergei/J-5936-2013; Nedospasov, Sergei/L-1990-2015; Nedospasov, Sergei/Q-7319-2016 FU NCI NIH HHS [N01-CO-56000] NR 32 TC 20 Z9 22 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 1466-4879 J9 GENES IMMUN JI Genes Immun. PD FEB PY 2000 VL 1 IS 3 BP 191 EP 199 DI 10.1038/sj.gene.6363659 PG 9 WC Genetics & Heredity; Immunology SC Genetics & Heredity; Immunology GA 342TX UT WOS:000088662200003 PM 11196711 ER PT J AU Eskdale, J Turestkaya, RL Armstrong, C Kuprash, DV Nedospasov, SA Gallagher, G AF Eskdale, J Turestkaya, RL Armstrong, C Kuprash, DV Nedospasov, SA Gallagher, G TI A polymorphic microsatellite marker in the human p55 TNF receptor, CD120a SO GENES AND IMMUNITY LA English DT Article DE microsatellite; TNF; TNF-R1; CD1ZOa; polymorphism ID NECROSIS-FACTOR RECEPTOR; DISEASE; DEATH; ROLES; ALPHA; GENE; P75 AB We provide characterization of a highly polymorphic dinucleotide repeat in the human TNF-receptor 1 gene (TNFR1, TNFRp55, CD120a). We have observed 11 alleles at this locus in individuals from the West of Scotland. in a panel of healthy, unrelated individuals from the West of Scotland (n = 143), the overall heterozygosity was 68%, indicating the potential usefulness of these markers in immunogenetic studies. C1 Univ Glasgow, Royal Infirm, Dept Surg, Glasgow G31 2ER, Lanark, Scotland. Russian Acad Sci, VA Engelhardt Mol Biol Inst, Lab Mol Immunol, Moscow 117984, Russia. Moscow MV Lomonosov State Univ, Belozersky Inst Physicochem Biol, Moscow 119899, Russia. SAIC Frederick, Intramural Res Support Program, Frederick, MD 21702 USA. NCI, Mol Immunoregulat Lab, Div Basic Sci, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RP Gallagher, G (reprint author), Univ Glasgow, Royal Infirm, Dept Surg, Queen Elizabeth Bldg, Glasgow G31 2ER, Lanark, Scotland. RI Nedospasov, Sergei/J-5936-2013; Nedospasov, Sergei/L-1990-2015; Kuprash, Dmitry/O-4899-2015; Nedospasov, Sergei/Q-7319-2016 OI Kuprash, Dmitry/0000-0002-1488-4148; FU NCI NIH HHS [N01-CO-56000] NR 16 TC 7 Z9 8 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 1466-4879 J9 GENES IMMUN JI Genes Immun. PD FEB PY 2000 VL 1 IS 3 BP 228 EP 230 DI 10.1038/sj.gene.6363663 PG 3 WC Genetics & Heredity; Immunology SC Genetics & Heredity; Immunology GA 342TX UT WOS:000088662200009 PM 11196717 ER PT J AU Gaidano, G Vivenza, D Forconi, F Capello, D Gloghini, A Bhatia, K Gutierrez, M Gallicchio, M Avanzi, GC Fassone, L Ariatti, C Buonaiuto, D Cingolani, A Saglio, G Tirelli, U Larocca, LM Dalla-Favera, R Carbone, A AF Gaidano, G Vivenza, D Forconi, F Capello, D Gloghini, A Bhatia, K Gutierrez, M Gallicchio, M Avanzi, GC Fassone, L Ariatti, C Buonaiuto, D Cingolani, A Saglio, G Tirelli, U Larocca, LM Dalla-Favera, R Carbone, A TI Mutation of BAX occurs infrequently in acquired immunodeficiency syndrome-related non-Hodgkin's lymphomas SO GENES CHROMOSOMES & CANCER LA English DT Article ID CAVITY-BASED LYMPHOMAS; EPSTEIN-BARR-VIRUS; IN-VITRO ESTABLISHMENT; TUMOR-SUPPRESSOR P53; CELL-LINES; DNA-SEQUENCES; B-CELLS; AIDS; GENE; HETEROGENEITY AB Acquired immunodeficiency syndrome (AIDS)-related non-Hodgkin's lymphomas (AIDS-NHLs) consistently derive from B cells, are histologically heterogeneous, and are associated with distinct molecular pathways depending upon histology. Recently, it has been proposed that inactivating mutations of the bax death agonist may contribute to the pathogenesis of human tumors. In particular, among B-cell malignancies, BAX mutations have been detected at a certain frequency in Burkitt lymphomas. This study is aimed at defining the status of the BAX gene throughout the clinicopathologic spectrum of AIDS-NHL (n = 54), including AIDS-related Burkitt lymphoma (n = 14), AIDS-related Burkitt-like lymphoma (n = 8), AIDS-related diffuse large cell lymphoma (n = 15), AIDS-related primary central nervous system lymphoma (n = 6), and AIDS-related primary effusion lymphoma (n =11). All 6 BAX exons and flanking sequences were subjected to mutational analysis by polymerase chain reaction-single strand conformation polymorphism followed by DNA direct sequencing of positive cases. Mutations of BAX among AIDS-NHL were restricted to a cell line of AIDS-related primary effusion lymphoma, which harbored a frameshift mutation causing the introduction of a proximal stop codon. All other AIDS-NHL displayed wild-type BAX alleles. In order to investigate whether BAX inactivation in AIDS-NHL may occur through mechanisms other than gene mutation, bax protein expression was investigated by Western blot analysis or immunohistochemistry in selected cases. All AIDS-NHL analyzed expressed normal bax proteins. Overall, this study indicates that deregulation of apoptotic control in AIDS-NHL is not caused by BAX alterations. Genes Chromosomes Cancer 27:177-182, 2000. (C) 2000 Wiley-Liss, Inc. C1 Amedeo Avogadro Univ Eastern Piedmont, Dept Med Sci, Div Internal Med, I-28100 Novara, Italy. Ist Nazl Tumori, Ctr Riferimento Oncol, Div Pathol, Aviano, Italy. NCI, Lymphoma Biol Sect, Pediat Branch, NIH, Bethesda, MD 20892 USA. Univ Sacred Heart, Inst Infect Dis, I-00168 Rome, Italy. Univ Turin, Dept Clin & Biol Sci, Div Internal Med & Hematol, Orbassano, Italy. Ist Nazl Tumori, Ctr Riferimento Oncol, Div Med Oncol A, Aviano, Italy. Univ Sacred Heart, Inst Pathol, I-00168 Rome, Italy. Columbia Univ, Coll Phys & Surg, Dept Pathol, Div Oncol, New York, NY USA. RP Gaidano, G (reprint author), Amedeo Avogadro Univ Eastern Piedmont, Dept Med Sci, Div Internal Med, Via Solaroli 17, I-28100 Novara, Italy. RI Larocca, Luigi/A-1577-2008; Capello, Daniela/J-4110-2012; OI Larocca, Luigi/0000-0003-1739-4758; Capello, Daniela/0000-0001-9157-8753; cingolani, antonella/0000-0002-3793-2755; Gloghini, Annunziata/0000-0002-7226-1942; Forconi, Francesco/0000-0002-2211-1831 NR 42 TC 11 Z9 11 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 1045-2257 J9 GENE CHROMOSOME CANC JI Gene Chromosomes Cancer PD FEB PY 2000 VL 27 IS 2 BP 177 EP 182 DI 10.1002/(SICI)1098-2264(200002)27:2<177::AID-GCC9>3.0.CO;2-O PG 6 WC Oncology; Genetics & Heredity SC Oncology; Genetics & Heredity GA 268TQ UT WOS:000084432000009 PM 10612806 ER PT J AU Ghadimi, BM Sackett, DL Difilippantonio, MJ Schrock, E Neumann, T Jauho, A Auer, G Ried, T AF Ghadimi, BM Sackett, DL Difilippantonio, MJ Schrock, E Neumann, T Jauho, A Auer, G Ried, T TI Centrosome amplification and instability occurs exclusively in aneuploid, but not in diploid colorectal cancer cell lines, and correlates with numerical chromosomal aberrations SO GENES CHROMOSOMES & CANCER LA English DT Article ID COMPARATIVE GENOMIC HYBRIDIZATION; GENETIC INSTABILITY; TUMORS; REVEALS; PLK1; POLO; P53 AB Measurement of the nuclear DNA content allows classification of human cancers as either diploid or aneuploid. To gain further insight into mechanisms of aneuploidy, we compared the cytogenetic profile of mismatch-repair-deficient diploid versus mismatch-repair-proficient aneuploid colorectal carcinoma cell lines using comparative genomic hybridization and spectral karyotyping. Aneuploid carcinomas revealed an average of 19 chromosomal imbalances per cell line. Such numerical aberrations were exceedingly scarce in the diploid tumors. This pattern of chromosomal aberrations is consistent with a mechanism involving the impairment of chromosome segregation fidelity during mitotic cell division. In support of this idea, we demonstrate the exclusive occurrence of centrosome amplification and instability in all of the aneuploid tumor cell lines analyzed. All diploid tumors contained centrosomes that were functionally and structurally indistinguishable from those in normal human fibroblasts. Due to the observed differences in centrosomes between these two classes of tumors, we incubated the cells with the microtubule depolymerizing drugs nocodazole and griseofulvin. Our results indicate that the aneuploid tumor cell lines have an increased sensitivity to these reagents and a delay in aster formation and microtubule regrowth. However, microtubule nucleation was initiated from one or two centers in both the diploid and aneuploid cells. These observations support the notion that the integrity of the centrosome plays a central role in the development of aneuploidy. Genes Chromosomes Cancer 27: 183-190, 2000. Published 2000 Wiley-Liss, Inc.dagger. C1 NCI, Dept Genet, NIH, Bethesda, MD 20892 USA. NCI, Lab Drug Discovery Res & Dev, NIH, Bethesda, MD 20892 USA. Karolinska Hosp & Inst, Dept Pathol & Oncol, Div Cellular & Mol Anal, Stockholm, Sweden. RP Ried, T (reprint author), NCI, Dept Genet, NIH, Bldg 49,Room 4A28,49 Convent Dr, Bethesda, MD 20892 USA. FU Intramural NIH HHS [Z99 CA999999] NR 19 TC 205 Z9 210 U1 0 U2 7 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 1045-2257 J9 GENE CHROMOSOME CANC JI Gene Chromosomes Cancer PD FEB PY 2000 VL 27 IS 2 BP 183 EP 190 DI 10.1002/(SICI)1098-2264(200002)27:2<183::AID-GCC10>3.0.CO;2-P PG 8 WC Oncology; Genetics & Heredity SC Oncology; Genetics & Heredity GA 268TQ UT WOS:000084432000010 PM 10612807 ER PT J AU de Vries, WN Binns, LT Fancher, KS Dean, J Moore, R Kemler, R Knowles, BB AF de Vries, WN Binns, LT Fancher, KS Dean, J Moore, R Kemler, R Knowles, BB TI Expression of Cre recombinase in mouse oocytes: A means to study maternal effect genes SO GENESIS LA English DT Article ID MESSENGER-RNA; POLYADENYLATION; UNMASKING; MASKING C1 Jackson Lab, Bar Harbor, ME 04609 USA. Wesleyan Univ, Middletown, CT USA. NIH, Bethesda, MD 20892 USA. Max Planck Inst Immunbiol, D-7800 Freiburg, Germany. Inst Curie, F-91405 Orsay, France. RP Knowles, BB (reprint author), Jackson Lab, 600 Main St, Bar Harbor, ME 04609 USA. FU NCI NIH HHS [P30 CA34186]; NICHD NIH HHS [HD37102, HD35252] NR 9 TC 165 Z9 166 U1 0 U2 2 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 1526-954X J9 GENESIS JI Genesis PD FEB PY 2000 VL 26 IS 2 BP 110 EP 112 DI 10.1002/(SICI)1526-968X(200002)26:2<110::AID-GENE2>3.0.CO;2-8 PG 3 WC Developmental Biology; Genetics & Heredity SC Developmental Biology; Genetics & Heredity GA 358KE UT WOS:000089556600002 PM 10686600 ER PT J AU Lowe, LA Yamada, S Kuehn, MR AF Lowe, LA Yamada, S Kuehn, MR TI HoxB6-Cre transgenic mice express Cre recombinase in extra-embryonic mesoderm, in lateral plate and limb mesoderm and at the midbrain/hindbrain junction SO GENESIS LA English DT Article ID REPORTER STRAIN; GENE C1 NCI, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Kuehn, MR (reprint author), NCI, Div Basic Sci, NIH, Bldg 10 Rm 4B-05,10 Ctr Dr, Bethesda, MD 20892 USA. RI Kuehn, Michael/A-4573-2014 OI Kuehn, Michael/0000-0002-7703-9160 NR 8 TC 25 Z9 25 U1 1 U2 2 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 1526-954X J9 GENESIS JI Genesis PD FEB PY 2000 VL 26 IS 2 BP 118 EP 120 DI 10.1002/(SICI)1526-968X(200002)26:2<118::AID-GENE5>3.0.CO;2-S PG 3 WC Developmental Biology; Genetics & Heredity SC Developmental Biology; Genetics & Heredity GA 358KE UT WOS:000089556600005 PM 10686603 ER PT J AU Rattray, AJ Shafer, BK Garfinkel, DJ AF Rattray, AJ Shafer, BK Garfinkel, DJ TI The Saccharomyces cerevisiae DNA recombination and repair functions of the RAD52 epistasis group inhibit Tyl transposition SO GENETICS LA English DT Review ID DOUBLE-STRAND BREAKS; NUCLEOTIDE EXCISION-REPAIR; REPLICATION PROTEIN-A; YEAST RETROTRANSPOSON TY; VIRUS-LIKE PARTICLES; MISMATCH REPAIR; MITOTIC RECOMBINATION; HOMOLOGOUS RECOMBINATION; REVERSE-TRANSCRIPTASE; MEIOTIC RECOMBINATION AB RNA transcribed from the Saccharomyces cerevisiae retrotransposon Ty1 accumulates to a high level in mitotically growing haploid cells, yet transposition occurs at very low frequencies. The product of reverse transcription is a linear double-stranded DNA molecule that reenters the genome by either Ty1-integrase-mediated insertion or homologous recombination with one of the preexisting genomic Ty1 (or delta) elements. Here we examine the role of the cellular homologous recombination functions on Ty1 transposition. We find that transposition is elevated in cells mutated for genes in the RAD52 recombinational repair pathway, such as RAD50, RAD51, RAD52 RAD54, or RAD57, or in the DNA ligase I gene CDC9, but is not elevated in cells mutated in the DNA repair functions encoded by the RAD1, RAD2, or MSH2 genes. The increase in Ty1 transposition observed when genes in the RAD52 recombinational pathway are mutated is not associated with a significant increase in Ty1 RNA or proteins. However, unincorporated Ty1 cDNA levels are markedly elevated. These results suggest that members of the RAD52 recombinational repair pathway inhibit Ty1 post-translationally by influencing the fate of Ty1 cDNA. C1 NCI, Gene Regulat & Chromosome Biol Lab, ABL Basic Res Program, FCRDC, Frederick, MD 21702 USA. RP Rattray, AJ (reprint author), NCI, Gene Regulat & Chromosome Biol Lab, ABL Basic Res Program, FCRDC, POB B,Bldg 539,Rm 151, Frederick, MD 21702 USA. RI Rattray, Alison/A-4847-2008 NR 135 TC 30 Z9 30 U1 0 U2 2 PU GENETICS PI BALTIMORE PA 428 EAST PRESTON ST, BALTIMORE, MD 21202 USA SN 0016-6731 J9 GENETICS JI Genetics PD FEB PY 2000 VL 154 IS 2 BP 543 EP 556 PG 14 WC Genetics & Heredity SC Genetics & Heredity GA 281UG UT WOS:000085178700006 PM 10655210 ER PT J AU O'Donnell, CJ AF O'Donnell, Christopher J. TI Cardiovascular genomics: recent progress, current challenges, future promise SO GENOME BIOLOGY LA English DT Editorial Material AB A report from the second Annual Cardiovascular Genomics Meeting, Sponsored by Cambridge Healthtech, Inc., Miami, January 31-February 1, 2000. C1 [O'Donnell, Christopher J.] Massachusetts Gen Hosp, Div Cardiol, Framingham, MA 02172 USA. [O'Donnell, Christopher J.] NHLBI, Framingham Heart Study, Framingham, MA 02172 USA. RP O'Donnell, CJ (reprint author), Massachusetts Gen Hosp, Div Cardiol, 5 Thurber St, Framingham, MA 02172 USA. EM chris@fram.nhlbi.nih.gov NR 0 TC 0 Z9 0 U1 0 U2 0 PU BIOMED CENTRAL LTD PI LONDON PA 236 GRAYS INN RD, FLOOR 6, LONDON WC1X 8HL, ENGLAND SN 1465-6906 EI 1474-760X J9 GENOME BIOL JI Genome Biol. PD FEB PY 2000 VL 1 IS 1 AR 409 PG 3 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA V34QL UT WOS:000209100800016 ER PT J AU Touchman, JW Anikster, Y Dietrich, NL Maduro, VVB McDowell, G Shotelersuk, V Bouffard, GG Beckstrom-Sternberg, SM Gahl, WA Green, ED AF Touchman, JW Anikster, Y Dietrich, NL Maduro, VVB McDowell, G Shotelersuk, V Bouffard, GG Beckstrom-Sternberg, SM Gahl, WA Green, ED TI The genomic region encompassing the nephropathic cystinosis gene (CTNS): Complete sequencing of a 200-kb segment and discovery of a novel gene within the common cystinosis-causing deletion SO GENOME RESEARCH LA English DT Article ID MYELOID-LEUKEMIA; DNA; COMPLICATIONS; INSUFFICIENCY; RECOMBINATION; MUTATIONS; RECEPTOR; SUBUNIT; CLONING; 17P13 AB Nephropathic cystinosis is an autosomal recessive disorder caused by the defective transport of cystine our of lysosomes. Recently, the causative gene (CTNS) was identified and presumed to encode an integral membrane protein called cystinosin. Many of the disease-associated mutations in CTNS are deletions, including one >55 kb in size that represents the most common cystinosis allele encountered to date. In an effort to determine the precise genomic organization of CTNS and to gain sequence-based insight about the DNA within and flanking cystinosis-associated deletions, we mapped and sequenced the region of human chromosome 17p13 encompassing CTNS. Specifically, a bacterial artificial chromosome (BAC)-based physical map spanning CTNS was constructed by sequence-tagged site (STS)-content mapping. The resulting BAC contig provided the relative order of 43 STSs. Two overlapping BACs, which together contain all of the CTNS exons as well as extensive amounts of flanking DNA, were selected and subjected to shotgun sequencing. A total of 200,237 bp of contiguous, high-accuracy sequence was generated. Analysis of the resulting data revealed a number of interesting features about this genomic region, including the long-range organization of CTNS insight about the breakpoints and intervening DNA associated with the common cystinosis-causing deletion, and structural information about Five genes neighboring CTNS (human ortholog of rat vanilloid receptor subtype 1 gene, CARKL, TIP-1, P2X5, and HUMINAE). In particular, sequence analysis detected the presence of a novel gene (CARKL) residing within the most common cystinosis-causing deletion. This gene encodes a previously unknown protein that is predicted to Function as a carbohydrate kinase. Interestingly, both CTNS and CARKL are absent in nearly half of all cystinosis patients (i.e., those homozygous For the common deletion). C1 NIH, NIH Intramural Sequencing Ctr, Gaithersburg, MD 20877 USA. NICHD, Heritable Disorders Branch, NIH, Bethesda, MD 20892 USA. Natl Human Genome Res Inst, Genome Technol Branch, NIH, Bethesda, MD 20892 USA. RP Green, ED (reprint author), NIH, NIH Intramural Sequencing Ctr, Gaithersburg, MD 20877 USA. NR 47 TC 77 Z9 86 U1 1 U2 5 PU COLD SPRING HARBOR LAB PRESS PI PLAINVIEW PA 1 BUNGTOWN RD, PLAINVIEW, NY 11724 USA SN 1088-9051 J9 GENOME RES JI Genome Res. PD FEB PY 2000 VL 10 IS 2 BP 165 EP 173 DI 10.1101/gr.10.2.165 PG 9 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 285ZH UT WOS:000085417900004 PM 10673275 ER PT J AU Santini, G Coser, P Congiu, AM Salvagno, L De Souza, C Sertoli, MR Olivieri, A Chisesi, T Rubagotti, A Truini, M Contu, A Porcellini, A Zambaldi, G Nati, S Marino, G Rizzoli, V AF Santini, G Coser, P Congiu, AM Salvagno, L De Souza, C Sertoli, MR Olivieri, A Chisesi, T Rubagotti, A Truini, M Contu, A Porcellini, A Zambaldi, G Nati, S Marino, G Rizzoli, V TI VACOP-B, high-dose cyclophosphamide and high-dose therapy with peripheral blood progenitor cell rescue for aggressive non-Hodgkin's lymphoma with bone marrow involvement: a study by the non-Hodgkin's Lymphoma Co-operative Study Group SO HAEMATOLOGICA LA English DT Article DE high-dose therapy; aggressive non-Hodgkin's lymphoma; bone marrow involvement ID COLONY-STIMULATING FACTOR; AUTOLOGOUS TRANSPLANTATION; ITALIAN MULTICENTER; MACOP-B; CHEMOTHERAPY; IDENTIFICATION; DEXAMETHASONE; MOBILIZATION; CISPLATIN; REGIMEN AB Background and Objectives. Sequential treatment with the addition of high-dose therapy (HDT) and peripheral blood progenitor cell (PBPC) rescue has been reported to be active as front-line therapy in aggressive non-Hodgkin's lymphoma (NHL) with bone marrow (BM) involvement. We designed an intensive sequential therapy as front-line therapy in this subset of patients and conducted a phase II study. Design and Methods. Patients with aggressive non-Hodgkin's lymphoma and BM involvement at diagnosis received 8 weeks of VACOP-B chemotherapy as Induction therapy. The second phase included high-dose cyclophosphamide (HDCY) (7 g/m(2)) with granulocyte colony-stimulating factor (G-CSF) followed by leukaphereses. The third phase included HDT according to the BEAM protocol or melphalan (140 mg/m(2)) plus total body irradiation (8 Gy in a single dose). Results. Forty patients were Included in the study. According to the intention-to-treat, after VACOP-B, 11 (27.5%) and 22 (55%) patients achieved complete remission (CR) and partial remission (PR), respectively. Thirty-four received HDCY. After HDCY, 18 patients (45%) were in CR and 13 (32.5%) in PR. Twenty-nine underwent HDT plus peripheral blood cell rescue (PBPC) rescue. At the completion of treatment 29 patients (72.5%) were in CR, and 3 patients (7.5%) in PR. The actuarial 3-year overall survival, disease free survival and failure free sur vival are 48%, 55% and 40%, respectively. Overall severe toxicity was 7.5%. Interpretation and Conclusions. This phase tl study suggests that the intensified treatment described is feasible and active in aggressive NHL with BM involvement. A randomized trial is now underway to test this approach. (C)2000, Ferrata Storti Foundation. C1 Osped San Martino Genova, Dept Hematol, Genoa, Italy. San Maurizio Hosp, Dept Hematol, Bolzano, Italy. Civil Hosp, Oncol Unit, Vittorio Veneto, Italy. Univ Estadual Campinas, BMT Unit, Hematol & Blood Transfus Ctr, BR-13081970 Campinas, SP, Brazil. Univ Genoa, Dept Oncol, Genoa, Italy. Univ Genoa, Biostat Unit, Genoa, Italy. Natl Canc Inst, Genoa, Italy. Univ Ancona, Le Torrette Hosp, Dept Hematol, Ancona, Italy. Venezia Hosp, Dept Hematol, Venice, Italy. Civil Hosp, Dept Med Oncol, Sassari, Italy. Ist Osped Cremonesi, Dept Hematol, Casalmaggiore, Cremona, Italy. Arco Hosp, Trento, Italy. Univ Parma, Dept Hematol, BMT Unit, I-43100 Parma, Italy. RP Santini, G (reprint author), Azienda Osped S Martino, Div Ematol 1, Largo R Benzi 10, I-16132 Genoa, Italy. NR 30 TC 9 Z9 12 U1 0 U2 1 PU FERRATA STORTI FOUNDATION PI PAVIA PA STRADA NUOVA 134, 27100 PAVIA, ITALY SN 0390-6078 J9 HAEMATOLOGICA JI Haematologica PD FEB PY 2000 VL 85 IS 2 BP 160 EP 166 PG 7 WC Hematology SC Hematology GA 286DX UT WOS:000085429300009 PM 10681723 ER PT J AU Yancik, R Ries, LAG AF Yancik, R Ries, LAG TI Cancer and aging in America - Demographic and epidemiologic perspectives SO HEMATOLOGY-ONCOLOGY CLINICS OF NORTH AMERICA LA English DT Article AB Data on cancer that emphasize the impact of this disease on older persons in the context of an expanding aging population in the United States is described using population-based statistics on incidence and mortality. Sixty percent of all incident cancers and 70% of all cancer mortality occur in people 65 years and older. A combined cancer and aging epiemiologic perspective of the cancer burden for the elderly is given, providing a backdrop for the articles in this issue. There is an urgent need for special attention to older persons as a target group for the integration of aging and cancer research. A discussion on pertinent clinical issues and questions that should be considered in treatment of cancer in the elderly is also provided. C1 NIA, NIH, Canc Sect, Geriatr Program, Bethesda, MD 20892 USA. NCI, Canc Stat Branch, Canc Surveillance Res Program, Div Canc Control & Populat Sci, Bethesda, MD USA. RP Yancik, R (reprint author), NIA, NIH, Canc Sect, Geriatr Program, Gateway Bldg,7201 Wisconsin Ave,Suite 3E327,MSC 9, Bethesda, MD 20892 USA. NR 9 TC 252 Z9 255 U1 1 U2 1 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0889-8588 J9 HEMATOL ONCOL CLIN N JI Hematol. Oncol. Clin. North Am. PD FEB PY 2000 VL 14 IS 1 BP 17 EP + DI 10.1016/S0889-8588(05)70275-6 PG 9 WC Oncology; Hematology SC Oncology; Hematology GA 282YF UT WOS:000085248800003 PM 10680069 ER PT J AU Ruhl, CE Everhart, JE AF Ruhl, CE Everhart, JE TI Association of diabetes, serum insulin, and C-peptide with gallbladder disease SO HEPATOLOGY LA English DT Article ID NON-HISPANIC WHITES; GALL STONE DISEASE; GALLSTONE DISEASE; RESISTANCE-ATHEROSCLEROSIS; GLUCOSE-TOLERANCE; PHYSICAL-ACTIVITY; FASTING INSULIN; HIGH PREVALENCE; PLASMA-INSULIN; YOUNG-ADULTS AB An inconsistent association has been found between gallbladder disease and diabetes mellitus. We hypothesized that insulin resistance rather than diabetes status may be a primary factor involved in gallstone formation. A total of 5,653 adult participants in the third United States National Health and Nutrition Examination Survey without known diabetes underwent gallbladder ultrasonography and phlebotomy after an overnight fast for measurement of serum insulin, C-peptide, and glucose. Gallbladder disease was defined as ultrasound-documented gallstones or evidence of cholecystectomy. Subjects were characterized as having normal fasting glucose (<110 mg/dL), impaired fasting glucose (110 to < 126 mg/dL), or undiagnosed diabetes (greater than or equal to 126 mg/dL). After controlling for other known gallbladder disease risk factors, among women, undiagnosed diabetes was associated with increased risk of gallbladder disease (prevalence ratio [PR] = 1.91, 95% confidence interval [CI] = 1.29-2.83); whereas impaired fasting glucose was unassociated, Gallbladder disease risk in women increased with levels of fasting insulin (PR = 1.63, 95% CI = 1.11-2.40) and C-peptide (PR = 2.07, 95% CI = 1.32-3.25) comparing highest to lowest quintiles. However, the association of gallbladder disease with undiagnosed diabetes was not diminished when the model included fasting insulin (PR = 1.85, 95% CI = 1.24-2.77), In men, there was a statistically nonsignificant association with undiagnosed diabetes (PR = 2.11, 95% CI = 0.76-5.85), but no association of gallbladder disease with insulin or C-peptide, Among women higher fasting serum insulin levels increased the risk of gallbladder disease, bur did not account for the increased risk in persons with diabetes. C1 Social & Sci Syst Inc, Bethesda, MD 20814 USA. NIDDKD, NIH, Bethesda, MD 20892 USA. RP Ruhl, CE (reprint author), Social & Sci Syst Inc, 7101 Wisconsin Ave,Suite 1300, Bethesda, MD 20814 USA. FU NIDDK NIH HHS [N01-DK-6-2220] NR 41 TC 91 Z9 98 U1 2 U2 3 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0270-9139 J9 HEPATOLOGY JI Hepatology PD FEB PY 2000 VL 31 IS 2 BP 299 EP 303 DI 10.1002/hep.510310206 PG 5 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 281EB UT WOS:000085144000006 PM 10655249 ER PT J AU Bergwitz, C Wendlandt, T Potter, E Glomb, I Gras, K von zur Muhlen, A Brabant, G AF Bergwitz, C Wendlandt, T Potter, E Glomb, I Gras, K von zur Muhlen, A Brabant, G TI A versatile chondrogenic rat calvaria cell line R-tTA-24 that permits tetracycline-regulated gene expression SO HISTOCHEMISTRY AND CELL BIOLOGY LA English DT Article ID CARTILAGE DIFFERENTIATION; PARATHYROID-HORMONE; BONE; DEXAMETHASONE; DOXYCYCLINE; POPULATION; INVITRO; MUSCLE AB The clonal rat calvaria cell line RCJ3.1C5.18 (RCJ) undergoes chondrogenic differentiation after long-term culture post confluence. To allow flexible genetic manipulation, a tetracycline-regulated gene expression system was established in this cell line. Treatment with tetracycline in operational doses does not affect the differentiation of RCJ cells with respect to the markers tested. After stable transfection with pUHD15.1 containing the tetracycline transactivator (tTA) in the presence of pTK-hyg for hygromycin selection, 28 clones were isolated and characterized for alcian blue staining of cartilage-specific proteoglycans and for collagen type II expression. Clone R-tTA-24 was selected on the basis of phenotype and displayed tetracycline-dependent down-regulation of luciferase activity (tet-OFF system) by two orders of magnitude (57-149-fold) after stable transfection with the reporter gene pBI-EGFP/luc. The novel, chondrogenic cell line R-tTA-24 may be stably transfected with various genes of interest for tetracycline-regulated gene expression using neomycin selection and may be a valuable tool to study the process of chondro-genic differentiation in vitro. C1 NICHD, Heritable Disorders Branch, NIH, Bethesda, MD 20892 USA. Med Hsch Hannover, Abt Klin Endokrinol, D-30625 Hannover, Germany. RP Bergwitz, C (reprint author), NICHD, Heritable Disorders Branch, NIH, Ctr Dr, Bethesda, MD 20892 USA. NR 32 TC 9 Z9 10 U1 0 U2 0 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0301-5564 J9 HISTOCHEM CELL BIOL JI Histochem. Cell Biol. PD FEB PY 2000 VL 113 IS 2 BP 145 EP 150 DI 10.1007/s004180050017 PG 6 WC Cell Biology; Microscopy SC Cell Biology; Microscopy GA 298WY UT WOS:000086163600009 PM 10766267 ER PT J AU Mitchison, NA Muller, B Segal, RM AF Mitchison, NA Muller, B Segal, RM TI Natural variation in immune responsiveness, with special reference to immunodeficiency and promoter polymorphism in class II MHC genes SO HUMAN IMMUNOLOGY LA English DT Article; Proceedings Paper CT Annual Scientific Meeting of the Australian-and-South-East-Asian-Tissue-Typing-Association (ASEATTA) CY 1998 CL NEW DELHI, INDIA SP Australian & South Asian Tissue Typing Assoc, DBT, DST, ICMR DE MHC; immunodeficiency; polymorphism; heterozygote; promoter ID COLLAGEN-INDUCED ARTHRITIS; LYMPHOPROLIFERATIVE DISEASE; FAS LIGAND; F-ANTIGEN; T-HELPER; EXPRESSION; CELLS; MICE; SUSCEPTIBILITY; INTERLEUKIN-4 AB This review deals with natural selection operating on heterozygotes as a key factor controlling (a) the frequency of immunodeficiencies, and (b) promoter polymorphism in MHC class II genes. The known difference in frequency distribution of X-linked and autosomal deficiencies lend support to this possibility, and suggest chat the frequency of neonatal defect may rise as old-established equilibria between entry and exit of deleterious mutations change. MHC class II gene promoters differ in their capacity to favor Th1 (or reciprocally Th2) responses, thus suggesting chat promoter polymorphism is sustained by the greater flexibility in response that this confers on heterozygotes. Human Immunology 61, 177-181 (2000). (C) American Society for Histocompatibility and Immunogenetics, 2000. Published by Elsevier Science Inc. C1 UCL, Sch Med, Dept Immunol, London W1P 6DB, England. Deutsch Rheuma Forsch Zentrum, D-10117 Berlin, Germany. NIH, Immunol Lab, Bethesda, MD 20892 USA. RP Mitchison, NA (reprint author), UCL, Sch Med, Dept Immunol, 46 Cleveland St, London W1P 6DB, England. EM n.mitchison@ucl.ac.uk RI mueller-hilke, brigitte/G-5583-2013 NR 34 TC 16 Z9 17 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 360 PARK AVE SOUTH, NEW YORK, NY 10010-1710 USA SN 0198-8859 J9 HUM IMMUNOL JI Hum. Immunol. PD FEB PY 2000 VL 61 IS 2 SI SI BP 177 EP 181 DI 10.1016/S0198-8859(99)00141-X PG 5 WC Immunology SC Immunology GA 279WC UT WOS:000085067200015 PM 10717812 ER PT J AU Aprile, MR Azzoni, C Gibril, F Jensen, RT Bordi, C AF Aprile, MR Azzoni, C Gibril, F Jensen, RT Bordi, C TI Intramucosal cysts in the gastric body of patients with Zollinger-Ellison syndrome SO HUMAN PATHOLOGY LA English DT Article DE gastric body; intramucosal cysts; Zollinger-Ellison syndrome ID SOMATOSTATIN RECEPTOR SCINTIGRAPHY; MULTIPLE ENDOCRINE NEOPLASIA; CLASSIFICATION; OMEPRAZOLE; PARIETAL; STOMACH; GROWTHS; THERAPY; POLYPS AB To ascertain the frequency and the clinico-functional correlations of intramucosal cysts in the gastric body of patients with the Zollinger-Ellison syndrome (ZES) and to clarify the relevant mechanism of development, a total of 106 consecutive ZES patients (58 M, 48 F; mean age: 53 yrs, range 19-93 yrs) were investigated with a mean of 7.2 biopsy specimens of the body mucosa per patient proved to be suitable for the study. Biopsies of endoscopically detectable polypoid lesions were not considered. Cystic changes were evaluated with respect to their severity by assessing the cyst grade (0, absent, 1; <30%, 2; 30-60%; 3 >60% of the mucosal area of the biopsy specimen of individual patients showing the most pronounced finding, respectively) and to their intragastric distribution by assessing the ratio of biopsy specimens showing cystic changes over the total number of biopsies examined in each patient. Intramucosal cysts were found in biopsies of non-polypoid gastric body mucosa in 71.7% of 106 patients with Zollinger-Ellison syndrome (ZES) and showed grade 2 and 3 severity in 22 and 8 cases, respectively. The severity of cystic changes correlated with the gastrin levels (p = 0.0005) and was more advanced in patients with active than in those with cured disease (p = 0.037). In the former group, furthermore, advanced cystic changes correlated with age (p = 0.03), male gender (p = 0.014), years of disease from onset (p < 0.02), years of omeprazole treatment (p = 0.033), basal acid output (p < 0.02), severity of ECL cell proliferative changes (p = 0.028), and absence of previous gastrinoma resection (p = 0.039) whereas they did not correlate with MEN-1 status, gastritis, maximal acid output, total duration of any antisecretory drug treatment, daily doses of omeprazole (>20 mg vs 20 mg), years from surgery, duodenal localization of gastrinoma(s), presence of gastric carcinoid tumor(s) and of liver metastases. In groups of patients subdivided according to three levels of serum gastrin, the duration of omeprazole treatment was not related to the severity of cystic changes. It is concluded that intramucosal cysts in non polypoid gastric body mucosa of ZES patients are by far more common than the already reported fundic gland polyps, to which they likely give raise. Circulating levels of gastrin have an important independent role in their development. Copyright (C) 2000 by W.E. Saunders Company. C1 Univ Parma, Dept Anat Pathol, I-43100 Parma, Italy. NIDDKD, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. RP Bordi, C (reprint author), Univ Parma, Ist Anat Patol, I-43100 Parma, Italy. NR 29 TC 17 Z9 17 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0046-8177 J9 HUM PATHOL JI Hum. Pathol. PD FEB PY 2000 VL 31 IS 2 BP 140 EP 148 DI 10.1016/S0046-8177(00)80213-0 PG 9 WC Pathology SC Pathology GA 282UR UT WOS:000085237900001 PM 10685627 ER PT J AU Otis, CN Krebs, PA Quezado, MM Albuquerque, A Bryant, B San Juan, X Kleiner, D Sobel, ME Merino, MJ AF Otis, CN Krebs, PA Quezado, MM Albuquerque, A Bryant, B San Juan, X Kleiner, D Sobel, ME Merino, MJ TI Loss of heterozygosity in P53, BRCA1, and estrogen receptor genes and correlation to expression of p53 protein in ovarian epithelial tumors of different cell types and biological behavior SO HUMAN PATHOLOGY LA English DT Article DE ovarian carcinoma; loss of heterozygosity; LOH; p53; BRCA1 ID MUTATION ANALYSIS; CANCER; CHROMOSOME-17; CARCINOMAS; BORDERLINE; OVEREXPRESSION; ALLELOTYPE; BREAST AB Loss of heterozygosity (LOH) of tumor suppressor genes (TSGs) in ovarian epithelial tumors of differing cell types and biological behavior has not been thoroughly investigated. Moreover, there have been conflicting reports correlating LOH of the p53 gene to overexpression of p53 protein. This study evaluated 34 formalin-fixed, paraffin-embedded ovarian epithelial tumors for LOH by polymerase chain reaction (PCR) for the following microsatellite markers: TP53(17p13.l/p53 gem), D17S579(17q/BRCA1 gene), and ESR (6q24-27/estrogen receptor gene). LOH of the TP53 marker was detected in 4 (44%) of 9 informative serous cystadenocarcinomas (SCa) but in 0 of 4 informative clear cell carcinomas (CCa) and 0 of 5 informative serous tumors of low malignant potential (SLMP). LOH of the BRCA1 marker was detected in 5 (83%) of 6 informative SCa, but in 1 (13%) of 8 informative CCa and 1 (14%) of 7 informative SLMP. LOH of the ESR marker was detected in 4 (50%) of 8 informative SCa, but in 0 of 4 informative CCa and 1 (16%) of 6 informative SLMP. p53 protein overexpression was present in 8 of 12 SCa but did not correlate to TP53 LOH. LOH for TP53, D17S579/BRCA1, and ESR is common in ovarian SCa, and is observed in primary tumors as well as metastases. In contrast, these genetic alterations are less common in CCa and in the biologically less aggressive SLMP tumors. These data suggest different mechanisms of oncogenesis in ovarian epithelial tumors of different cell types and biological behavior. Copyright (C) 2000 by W.B. Saunders Company. C1 Tufts Univ, Sch Med, Baystate Med Ctr, Dept Pathol, Springfield, MA 01199 USA. NCI, Surg Pathol Sect, NIH, Bethesda, MD 20892 USA. NCI, Mol Pahol Sect, NIH, Bethesda, MD 20892 USA. RP Otis, CN (reprint author), Tufts Univ, Sch Med, Baystate Med Ctr, Dept Pathol, Springfield, MA 01199 USA. OI Kleiner, David/0000-0003-3442-4453 NR 26 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 USA SN 0046-8177 J9 HUM PATHOL JI Hum. Pathol. PD FEB PY 2000 VL 31 IS 2 BP 233 EP 238 DI 10.1016/S0046-8177(00)80225-7 PG 6 WC Pathology SC Pathology GA 282UR UT WOS:000085237900013 PM 10685639 ER PT J AU Jarai, Z Wagner, JA Goparaju, SK Wang, L Razdan, RK Sugiura, T Zimmer, AM Bonner, TI Zimmer, A Kunos, G AF Jarai, Z Wagner, JA Goparaju, SK Wang, L Razdan, RK Sugiura, T Zimmer, AM Bonner, TI Zimmer, A Kunos, G TI Cardiovascular effects of 2-arachidonoyl glycerol in anesthetized mice SO HYPERTENSION LA English DT Article DE receptors; mice; hypotension; cannabinoid ID CANNABINOID CB1 RECEPTOR; SELECTIVE ANTAGONIST; INDUCED HYPOTENSION; ANANDAMIDE; BRAIN; CELLS; AGONIST; LIGAND; POTENT; BINDS AB Cannabinoids, including the endogenous ligand anandamide, elicit pronounced hypotension and bradycardia through the activation of CB1 cannabinoid receptors. A second endogenous cannabinoid, 2-arachidonoyl glycerol (2-AG), has been proposed to be the natural Ligand of CB1 receptors. In the present study, we examined the effects of 2-AG on mean arterial pressure and heart rate in anesthetized mice and assessed the role of CB1 receptors through the use of selective cannabinoid receptor antagonists and CB1 receptor knockout (CB1(-/-)) mice. In control ICR mice, intravenous injections of 2-AG or its isomer 1-AG elicit dose-dependent hypotension and moderate tachycardia that are unaffected by the CB1 receptor antagonist SR141716A. The same dose of SR141716A (6 nmol/g IV) completely blocks the hypotensive effect and attenuates the bradycardic effect of anandamide. 2-AG elicits a similar hypotensive effect, resistant to blockade by either SR141716A or the CB2 antagonist SR144528, in both CB1(-/-) mice and their homozygous (CB1(+/+)) control littermates. In ICR mice, arachidonic acid (AA, 15 nmol/g TV) elicits hypotension and tachycardia, and indomethacin (14 nmol/g IV) inhibits the hypotensive effect of both AA and 2-AG. Synthetic 2-AG incubated with mouse blood is rapidly (<2 minutes) and completely degraded with the parallel appearance of AA, whereas anandamide is stable under the same conditions. A metabolically stable ether analogue of 2-AG causes prolonged hypotension and bradycardia in ICR mice, and both effects are completely blocked by SR141716A, whereas the same dose of ZAG-ether does not influence blood pressure and heart rate in CB1(-/-) mice. These findings are interpreted to indicate that exogenous 2-AG is rapidly degraded in mouse blood, probably by a lipase, which masks its ability to interact with CB1 receptors. Although the observed cardiovascular effects of 2-AG probably are produced by an arachidonate metabolite through a noncannabinoid mechanism, the CB1 receptor-mediated cardiovascular effects of a stable analogue of 2-AG leaves open the possibility that endogenous 2-AG may elicit cardiovascular effects through CB1 receptors. C1 Virginia Commonwealth Univ, Med Coll Virginia, Dept Pharmacol & Toxicol, Richmond, VA 23298 USA. Organix Inc, Woburn, MA USA. Teikyo Univ, Fac Pharmaceut Sci, Kanagawa, Japan. NIMH, Genet Sect, Bethesda, MD 20892 USA. Univ Wurzburg, Dept Med, Wurzburg, Germany. RP Kunos, G (reprint author), Virginia Commonwealth Univ, Med Coll Virginia, Dept Pharmacol & Toxicol, 410 N 12th St,POB 980613, Richmond, VA 23298 USA. RI Goparaju, Sravan/H-3422-2011; Zimmer, Andreas/B-8357-2009 FU NHLBI NIH HHS [HL-59257] NR 31 TC 64 Z9 66 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0194-911X J9 HYPERTENSION JI Hypertension PD FEB PY 2000 VL 35 IS 2 BP 679 EP 684 PG 6 WC Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 285YA UT WOS:000085414900026 PM 10679517 ER PT J AU Singh, J Larson, M Levy, D AF Singh, J Larson, M Levy, D TI Heart rate variability: How to assess effects of mild therapies on autonomic control in small groups of mild and borderline hypertensives? Response SO HYPERTENSION LA English DT Letter C1 NHLBI, Framingham Heart Study, Framingham, MA 01702 USA. Boston Univ, Sch Med, Boston, MA 02118 USA. Harvard Univ, Massachusetts Gen Hosp, Sch Med, Boston, MA USA. RP Singh, J (reprint author), NHLBI, Framingham Heart Study, Framingham, MA 01702 USA. NR 4 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0194-911X J9 HYPERTENSION JI Hypertension PD FEB PY 2000 VL 35 IS 2 BP E7 EP E7 PG 1 WC Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 285YA UT WOS:000085414900030 ER PT J AU Lu, FWM Yasutomo, K Goodman, GB McHeyzer-Williams, LJ McHeyzer-Williams, MG Germain, RN Ashwell, JD AF Lu, FWM Yasutomo, K Goodman, GB McHeyzer-Williams, LJ McHeyzer-Williams, MG Germain, RN Ashwell, JD TI Thymocyte resistance to glucocorticoids leads to antigen-specific unresponsiveness due to "holes" in the T cell repertoire SO IMMUNITY LA English DT Article ID POSITIVE SELECTION; CYTOCHROME-C; RECEPTOR; MACROPHAGE; APOPTOSIS; COMPLEX; THYMUS; MODEL AB We have proposed that glucocorticoids antagonize TCR-mediated activation and influence which TCR avidities result in positive or negative selection. We now analyze the immune response of mice whose thymocytes express antisense transcripts to the glucocorticoid receptor (TKO mice). TKO mice responded normal ly to the complex antigen PPD but were proliferative nonresponders to pigeon cytochrome c 81-104 (PCC), having a large decrease in the frequency of PCC-responsive CD4(+) T cells. Unlike wild-type T cells, few TKO T cells in PCC-specific cell lines expressed V alpha 11(+)V beta 3(+). Furthermore, for naive CD4(+) T cells from unimmunized TKO mice, the frequencies of many of the molecular features common to the CDR3 regions of PCC-responsive V alpha 11(+)V beta 3(+) cells were substantially decreased. Thus, thymocyte glucocorticoid hyporesponsiveness resulted in loss of cells capable of responding to PCC, corresponding to an antigen-specific "hole" in the T cell repertoire. C1 NCI, Lab Immune Cell Biol, NIH, Bethesda, MD 20892 USA. NIAID, Lymphocyte Biol Sect, Immunol Lab, NIH, Bethesda, MD 20892 USA. Duke Univ, Med Ctr, Dept Immunol, Durham, NC 27710 USA. RP Ashwell, JD (reprint author), NCI, Lab Immune Cell Biol, NIH, Bethesda, MD 20892 USA. EM jda@box-j.nih.gov NR 23 TC 51 Z9 51 U1 0 U2 0 PU CELL PRESS PI CAMBRIDGE PA 600 TECHNOLOGY SQUARE, 5TH FLOOR, CAMBRIDGE, MA 02139 USA SN 1074-7613 J9 IMMUNITY JI Immunity PD FEB PY 2000 VL 12 IS 2 BP 183 EP 192 DI 10.1016/S1074-7613(00)80171-5 PG 10 WC Immunology SC Immunology GA 289LA UT WOS:000085621400008 PM 10714684 ER PT J AU Mekori, YA Metcalfe, DD AF Mekori, YA Metcalfe, DD TI Mast cells in innate immunity SO IMMUNOLOGICAL REVIEWS LA English DT Review ID TUMOR-NECROSIS-FACTOR; MACROPHAGE INFLAMMATORY PROTEIN-1-ALPHA; FC-EPSILON-RI; ADHESION MOLECULE-1; FACTOR-ALPHA; TNF-ALPHA; T-CELLS; SCHISTOSOMA-MANSONI; CYTOKINE PRODUCTION; ENDOTHELIAL-CELLS AB Masts cells are known to be the main effector cells in the elicitation of the IgE-mediated allergic response. The specific location of mast cells within tissues that interface the external environment, and the extent of their functional capacity, including the ability to phagocytose and to produce and secrete a wide spectrum of mediators, have led investigators to propose a potential role for mast cells in innate immune responses. Certain microorganisms have been found to interact either directly or indirectly with mast cells. This interaction results in mast cell activation and mediator release which elicit an inflammatory response or direct killing leading to bacterial clearance. The in vivo relevance of these in vitro observations has been demonstrated by the use of complement-deficient and/or mast cell-deficient and mast cell-reconstituted mice. It thus has been shown that both C3 and mast cell- and tumor necrosis factor-alpha-dependent recruitment of circulating leukocytes with bactericidal properties are crucial to a full response in certain models of acute infection. Modulation of mast cell numbers in vivo was also found to affect the host response against bacterial infection. Thus, mast cells do have a role in innate immunity in defined animal models of bacterial infection. Whether mast cells participate in innate immune responses in the protection of the human host against bacteria remains to be determined. C1 Meir Gen Hosp, Dept Med B, IL-44281 Kfar Saba, Israel. Tel Aviv Univ, Sackler Sch Med, IL-69978 Tel Aviv, Israel. NIAID, Lab Allerg Dis, NIH, Bethesda, MD USA. RP Mekori, YA (reprint author), Meir Gen Hosp, Dept Med B, IL-44281 Kfar Saba, Israel. NR 80 TC 261 Z9 266 U1 1 U2 7 PU MUNKSGAARD INT PUBL LTD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0105-2896 J9 IMMUNOL REV JI Immunol. Rev. PD FEB PY 2000 VL 173 BP 131 EP 140 DI 10.1034/j.1600-065X.2000.917305.x PG 10 WC Immunology SC Immunology GA 285WL UT WOS:000085411300012 PM 10719674 ER PT J AU Nilsson, G Metcalfe, DD Taub, DD AF Nilsson, G Metcalfe, DD Taub, DD TI Demonstration that platelet-activating factor is capable of activating mast cells and inducing a chemotactic response SO IMMUNOLOGY LA English DT Article ID SIGNAL-TRANSDUCTION PATHWAY; TRANSFORMING GROWTH-FACTOR-BETA-1; PEPTIDE CHEMOATTRACTANTS; CROSS-DESENSITIZATION; MEDIATOR RELEASE; RECEPTORS; ASTHMA; C5A; ANTAGONISTS; NEUTROPHILS AB Platelet-activating factor (PAF) is generated in a variety of inflammatory conditions in which mast cells accumulate. However, little is known about the ability of PAF to influence mast cell function directly. In this study we examine the ability of PAF to activate mast cells and regulate mast cell chemotaxis. PAF was found to induce intracellular calcium mobilization and chemotactic responses in both murine and human mast cells. PAF induced transient increases in intracellular Ca2+ concentrations with a 50% effective dose of 1 nm and induced significant migratory responses at PAF concentrations of 1 nm to 1 mu m in the human leukaemia mast cell line (HMC-1). Using signal transduction inhibitors, both PAF-induced calcium mobilization and migration of mast cells were shown to require activation of pertussis toxin-sensitive G proteins. PAF-induced calcium and chemotactic responses were cross-desensitized by C5a. Together, these data demonstrate that PAF is capable of activating distinct signalling pathways in mast cells associated with calcium mobilization and cell migration; and that PAF may thus contribute to the regulation of mast cell responses and hyperplasia at sites of inflammation. C1 Univ Uppsala, Dept Genet & Pathol, S-75185 Uppsala, Sweden. NIAID, Lab Allerg Dis, NIH, Bethesda, MD 20892 USA. NIA, Clin Immunol Sect, Immunol Lab, NIH, Baltimore, MD 21224 USA. RP Nilsson, G (reprint author), Univ Uppsala, Dept Genet & Pathol, S-75185 Uppsala, Sweden. NR 37 TC 42 Z9 43 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0019-2805 J9 IMMUNOLOGY JI Immunology PD FEB PY 2000 VL 99 IS 2 BP 314 EP 319 DI 10.1046/j.1365-2567.2000.00972.x PG 6 WC Immunology SC Immunology GA 289MM UT WOS:000085625600021 PM 10692052 ER PT J AU Puel, A Leonard, WJ AF Puel, A Leonard, WJ TI Interleukin-7 receptor alpha chain-dependent signaling is required for T-cell development - Basis for T-B+NK+ severe combined immunodeficiencies in humans SO IMMUNOLOGY AND ALLERGY CLINICS OF NORTH AMERICA LA English DT Article ID COMMON GAMMA-CHAIN; NATURAL-KILLER-CELLS; DEFECTIVE LYMPHOID DEVELOPMENT; JAK-3 JANUS KINASE; MICE LACKING JAK3; IL-2 RECEPTOR; DEFICIENT MICE; MURINE INTERLEUKIN-7; GROWTH-FACTOR; BETA-CHAIN AB X-linked SCID (XSCID) is characterized by profoundly diminished T and natural killer (NK) cells. XSCID is caused by mutations in the common cytokine receptor gamma chain, gamma(c), which is a component of the receptors for IL-2, IL-4, IL-7, IL-9, and IL-15. A similar clinical and immunologic spectrum results from mutations in JAK3, which associates with gamma(c). This article reviews these findings, and summarizes more recent evidence that mutations in IL-7R alpha cause a form of SCID in which T-tell numbers diminished profoundly but NK cells were normal. These findings not only provide the first elucidation of a basis for T-B+NK+ SCID but also suggest that defective IL-7R alpha-dependent signaling accounts for the T-cell defect in XSCID and JAK3 deficiency. Other implications of these findings also are discussed. C1 NHLBI, Mol Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Leonard, WJ (reprint author), NHLBI, Mol Immunol Lab, NIH, Bldg 10,Room 7N252,900 Rockville Pike, Bethesda, MD 20892 USA. NR 99 TC 5 Z9 5 U1 0 U2 2 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0889-8561 J9 IMMUNOL ALLERGY CLIN JI Immunol. Allerg. Clin. North Am. PD FEB PY 2000 VL 20 IS 1 BP 51 EP + DI 10.1016/S0889-8561(05)70133-8 PG 14 WC Allergy; Immunology SC Allergy; Immunology GA 283FK UT WOS:000085265300005 ER PT J AU Notarangelo, LD Candotti, F AF Notarangelo, LD Candotti, F TI JAK3-deficient severe combined immunodeficiency SO IMMUNOLOGY AND ALLERGY CLINICS OF NORTH AMERICA LA English DT Article ID RECEPTOR-GAMMA-CHAIN; DEFECTIVE LYMPHOID DEVELOPMENT; BONE-MARROW TRANSPLANTATION; JAK-3 JANUS KINASE; MICE LACKING JAK3; TYROSINE PHOSPHORYLATION; IL-2 RECEPTOR; MYELOID CELLS; T-CELLS; INTERLEUKIN-2 AB Genetic mutations affecting JAK3 expression or function result in autosomal recessive SCID. Children affected with JAK3-deficient SCID present with virtual absence of T lymphocytes and NK cells, whereas B lymphocytes usually are normal or elevated in number but functionally defective. Because of the profound impairment of cellular and humoral immunity, patients with JAK3 deficiency are exposed to recurrent and life-threatening infections unless immune reconstitution is achieved by allogeneic bone marrow transplantation. The identification of JAK3 as a key player in the development of the immune system in humans has stimulated many studies detailing the biologic characteristics of this important molecule, and has opened the way to possible genetic intervention. C1 Univ Brescia, Ist Med Mol Angelo Nocivelli, Dept Pediat, Spedali Civili, I-25123 Brescia, Italy. NHGRI, Clin Gene Therapy Branch, NIH, Bethesda, MD USA. RP Notarangelo, LD (reprint author), Univ Brescia, Ist Med Mol Angelo Nocivelli, Dept Pediat, Spedali Civili, I-25123 Brescia, Italy. RI Notarangelo, Luigi/F-9718-2016 OI Notarangelo, Luigi/0000-0002-8335-0262 NR 57 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 USA SN 0889-8561 J9 IMMUNOL ALLERGY CLIN JI Immunol. Allerg. Clin. North Am. PD FEB PY 2000 VL 20 IS 1 BP 97 EP + DI 10.1016/S0889-8561(05)70136-3 PG 16 WC Allergy; Immunology SC Allergy; Immunology GA 283FK UT WOS:000085265300008 ER PT J AU Ferrone, S Finerty, JF Jaffee, EM Nabel, GJ AF Ferrone, S Finerty, JF Jaffee, EM Nabel, GJ TI How much longer will tumour cells fool the immune system? SO IMMUNOLOGY TODAY LA English DT Article ID LIGAND AB In spite of successful control of tumour growth in mice; immunotherapy has yielded disappointing results in patients with malignant diseases; A recent meeting* focused on the mechanisms used by tumours to escape immune recognition and the implications for cancer immunotherapy. C1 Roswell Pk Canc Inst, Dept Immunol, Buffalo, NY 14263 USA. NCI, Canc Immunol & Hematol Branch, Rockville, MD 20852 USA. Johns Hopkins Univ, Sch Med, Dept Oncol & Immunol, Baltimore, MD 21205 USA. NIH, Vaccine Res Ctr, Bethesda, MD 20892 USA. RP Ferrone, S (reprint author), Roswell Pk Canc Inst, Dept Immunol, Elm & Carlton St, Buffalo, NY 14263 USA. NR 8 TC 37 Z9 40 U1 0 U2 1 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 0167-5699 J9 IMMUNOL TODAY JI Immunol. Today PD FEB PY 2000 VL 21 IS 2 BP 70 EP 72 DI 10.1016/S0167-5699(99)01569-8 PG 3 WC Immunology SC Immunology GA 281DA UT WOS:000085141800005 PM 10712001 ER PT J AU Nakajima, H Noguchi, M Leonard, WJ AF Nakajima, H Noguchi, M Leonard, WJ TI Role of-the common cytokine receptor gamma chain (gamma(c)) in thymocyte selection SO IMMUNOLOGY TODAY LA English DT Article ID SEVERE COMBINED IMMUNODEFICIENCY; T-CELL DEVELOPMENT; NEGATIVE SELECTION; POSITIVE SELECTION; DEFICIENT MICE; IL-7 RECEPTOR; STEM-CELLS; CALPAIN; TCR; INTERLEUKIN-7 AB During thymocyte development,, T-cell receptor (TCR) alpha beta-mediated intracellular signals can elicit two entirely different cellular responses: positive selection (resulting in rescue from death and maturation or differentiation) and negative selection (induction of apoptosis). Here, Hiroshi Nakajima and colleagues discuss how survival signals that ave dependent on the common cytokine receptor gamma drain (gamma(c)) might affect the TCR-driven selection process in thymocytes, underscoring the potential role of cytokines in this process. C1 Chiba Univ, Sch Med, Dept Internal Med 2, Chiba 2608670, Japan. Harvard Univ, Sch Med, Beth Israel Deaconess Med Ctr, Dept Med,Div Immunol, Boston, MA 02115 USA. NHLBI, Lab Mol Immunol, Bethesda, MD 20892 USA. RP Nakajima, H (reprint author), Chiba Univ, Sch Med, Dept Internal Med 2, Chiba 2608670, Japan. NR 52 TC 20 Z9 20 U1 0 U2 1 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 0167-5699 J9 IMMUNOL TODAY JI Immunol. Today PD FEB PY 2000 VL 21 IS 2 BP 88 EP 94 DI 10.1016/S0167-5699(99)01555-8 PG 7 WC Immunology SC Immunology GA 281DA UT WOS:000085141800009 PM 10652467 ER PT J AU Lukomski, S Hoe, NP Abdi, I Rurangirwa, J Kordari, P Liu, MY Dou, SJ Adams, GG Musser, JM AF Lukomski, S Hoe, NP Abdi, I Rurangirwa, J Kordari, P Liu, MY Dou, SJ Adams, GG Musser, JM TI Nonpolar inactivation of the hypervariable streptococcal inhibitor of complement gene (sic) in serotype M1 Streptococcus pyogenes significantly decreases mouse mucosal colonization SO INFECTION AND IMMUNITY LA English DT Article ID GROUP-A STREPTOCOCCUS; ESCHERICHIA-COLI; PROTEIN; CLONING; VIRULENCE; VECTORS; STRAINS; TN916; EXPRESSION; INFECTIONS AB Group A Streptococcas (GAS) is a human pathogen that commonly infects the upper respiratory tract. GAS serotype M1 strains are frequently isolated from human infections and contain the gene encoding the hypervariable streptococcal inhibitor of complement protein (Sic), It was recently shown that Sic variants were rapidly selected on mucosal surfaces in epidemic waves caused by M1 strains, an observation suggesting that Sic participates in host-pathogen interactions on the mucosal surface (N. P. Hoe, K. Nakashima, S, Lukomski, D. Grigsby, M. Liu, P, Kordari, S.-J. Dou, X. Pan, J. Vuopio-Varkila, S. Salmelinna, A. McGeer, D. E. Low, B. Schwartz, A. Schuchat, S, Naidich, D, De Lorenzo, Y.-X. Fu, and J, M, Musser, Nat, Med. 5:924-929, 1999), To test this idea, a new nonpolar mutagenesis method employing a spectinomycin resistance cassette was used to inactivate the sk gene in an MI GAS strain, The isogenic Sie-negative mutant strain was significantly (P < 0.019) impaired in ability to colonize the mouse mucosal surface after intranasal infection. These results support the hypothesis that the predominance of M1 strains in human infections is related, in part, to a Sie-mediated enhanced colonization ability. C1 NIAID, Rocky Mt Labs, Lab Human Bacterial Pathogenesis, NIH, Hamilton, MT 59840 USA. Baylor Coll Med, Dept Pathol, Inst Study Human Bacterial Pathogenesis, Houston, TX 77030 USA. RP Musser, JM (reprint author), NIAID, Rocky Mt Labs, Lab Human Bacterial Pathogenesis, NIH, 903 S 4th St, Hamilton, MT 59840 USA. FU NIAID NIH HHS [AI33119] NR 32 TC 96 Z9 98 U1 1 U2 2 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD FEB PY 2000 VL 68 IS 2 BP 535 EP 542 DI 10.1128/IAI.68.2.535-542.2000 PG 8 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 275VZ UT WOS:000084842000015 PM 10639414 ER PT J AU Rathore, D McCutchan, TF AF Rathore, D McCutchan, TF TI The cytotoxic T-lymphocyte epitope of the Plasmodium falciparum circumsporozoite protein also modulates the efficiency of receptor-ligand interaction with hepatocytes SO INFECTION AND IMMUNITY LA English DT Article ID HEPARAN-SULFATE PROTEOGLYCANS; MALARIA PARASITES CONTAIN; REGION-II-PLUS; ORAL SALMONELLA; THROMBOSPONDIN; SPOROZOITES; VACCINE; CELLS; SEQUENCE; GENE AB Malaria sporozoites are transmitted from the mosquito salivary gland to host hepatocytes within minutes of an infectious bite. The circumsporozoite protein (CS), which covers the surface of Plasmodium sporozoites, functions during these minutes in the targeting of host liver cells. The protein's potentially important role in an antimalaria vaccine has spawned interest in both the host immune responses to the parasite's presence and the actual functional role of the protein in the targeting of host liver cells. Here we show that the region of CS known to elicit a cytotoxic T-lymphocyte (CTL) response to irradiated sporozoites also, somewhat ironically, mediates the receptor-ligand interaction essential to parasite invasion of the host. Hence, the structure of CS represents a balance of potentially counterdirectional forces. Polymorphism in the CTL epitope appears to be a product of this balanced state as opposed to an "arms race" as it is so often portrayed. The conceptual difference between the theories regarding the maintainance of polymorphism in CTL epitopes may have significant implication for vaccine design. C1 NIAID, Parasit Dis Lab, Growth & Dev Sect, NIH, Bethesda, MD 20892 USA. RP McCutchan, TF (reprint author), NIAID, Parasit Dis Lab, Growth & Dev Sect, NIH, Rm 126,Bldg 4,4 Ctr Dr MSC 0425, Bethesda, MD 20892 USA. NR 40 TC 12 Z9 12 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD FEB PY 2000 VL 68 IS 2 BP 740 EP 743 DI 10.1128/IAI.68.2.740-743.2000 PG 4 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 275VZ UT WOS:000084842000042 PM 10639441 ER PT J AU George, KM Pascopella, L Welty, DM Small, PLC AF George, KM Pascopella, L Welty, DM Small, PLC TI A Mycobacterium ulcerans toxin, mycolactone, causes apoptosis in guinea pig ulcers and tissue culture cells SO INFECTION AND IMMUNITY LA English DT Article ID DNA FRAGMENTATION; EPITHELIAL-CELLS; ESCHERICHIA-COLI; DEATH; TUBERCULOSIS; LYMPHOCYTES; ACTIVATION; INHIBITION; INDUCTION; ABSENCE AB Mycobacterium ulcerans is the causative agent of Buruli ulcer, a tropical ulcerative skin disease. One of the most intriguing aspects of this disease is the presence of extensive tissue damage in the absence of an acute inflammatory response. We recently purified and characterized a macrolide toxin, mycolactone, from M. ulcerans. Injection of this molecule into guinea pig skin reproduced cell death and lack of acute inflammatory response similar to that seen following the injection of viable bacteria. We also showed that mycolactone causes a cytopathic effect on mouse fibroblast L929 cells that is characterized by cytoskeletal rearrangements and growth arrest within 48 h. However, these results could not account for the extensive cell death which occurs in Buruli ulcer. The results presented here demonstrate that L929 and J774 mouse macrophage cells die via apoptosis after 3 to 5 days of exposure to mycolactone. Treatment of cells with a pan-caspase inhibitor can inhibit mycolactone-induced apoptosis, We demonstrate that injection of mycolactone into guinea pig skin results in cell death via apoptosis and that the extent of apoptosis increases as the lesion progresses, These results may help to explain why tissue damage in Buruli ulcer is not accompanied by an acute inflammatory response. C1 NIAID, Rocky Mt Labs, Microscopy Branch, NIH, Hamilton, MT 59840 USA. RP Small, PLC (reprint author), NIAID, Rocky Mt Labs, Microscopy Branch, NIH, 903 S 4th St, Hamilton, MT 59840 USA. NR 32 TC 147 Z9 147 U1 0 U2 5 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 USA SN 0019-9567 J9 INFECT IMMUN JI Infect. Immun. PD FEB PY 2000 VL 68 IS 2 BP 877 EP 883 DI 10.1128/IAI.68.2.877-883.2000 PG 7 WC Immunology; Infectious Diseases SC Immunology; Infectious Diseases GA 275VZ UT WOS:000084842000059 PM 10639458 ER PT J AU Miele, PS Levy, CS Marcus, KA Rosenthal, J Gill, VJ Croxton, M Lucey, DR AF Miele, PS Levy, CS Marcus, KA Rosenthal, J Gill, VJ Croxton, M Lucey, DR TI Seven cases of native valve endocarditis due to coagulase negative staphylococci: A one year experience. SO INFECTION CONTROL AND HOSPITAL EPIDEMIOLOGY LA English DT Meeting Abstract C1 Kaiser Permanente, Washington, DC USA. Washington Hosp Ctr, Washington, DC 20010 USA. NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU SLACK INC PI THOROFARE PA 6900 GROVE RD, THOROFARE, NJ 08086 USA SN 0899-823X J9 INFECT CONT HOSP EP JI Infect. Control Hosp. Epidemiol. PD FEB PY 2000 VL 21 IS 2 BP 96 EP 96 PG 1 WC Public, Environmental & Occupational Health; Infectious Diseases SC Public, Environmental & Occupational Health; Infectious Diseases GA 285XM UT WOS:000085413700072 ER PT J AU Fahey, BJ Lee, LM Wesley, RA Henderson, DK AF Fahey, BJ Lee, LM Wesley, RA Henderson, DK TI Sustained decrease of cutaneous exposures among healthcare workers. SO INFECTION CONTROL AND HOSPITAL EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU SLACK INC PI THOROFARE PA 6900 GROVE RD, THOROFARE, NJ 08086 USA SN 0899-823X J9 INFECT CONT HOSP EP JI Infect. Control Hosp. Epidemiol. PD FEB PY 2000 VL 21 IS 2 BP 108 EP 109 PG 2 WC Public, Environmental & Occupational Health; Infectious Diseases SC Public, Environmental & Occupational Health; Infectious Diseases GA 285XM UT WOS:000085413700155 ER PT J AU Suzuki, K Nakajima, H Saito, Y Saito, T Leonard, WJ Iwamoto, I AF Suzuki, K Nakajima, H Saito, Y Saito, T Leonard, WJ Iwamoto, I TI Janus kinase 3 (Jak3) is essential for common cytokine receptor gamma chain (gamma(c))-dependent signaling: comparative analysis of gamma(c), Jak3, and gamma(c) and Jak3 double-deficient mice SO INTERNATIONAL IMMUNOLOGY LA English DT Article DE apoptosis; Bcl-2; common cytokine receptor gamma chain; Jak3; knockout mice ID SEVERE COMBINED IMMUNODEFICIENCY; DEFECTIVE LYMPHOID DEVELOPMENT; TRANSDUCING ADAPTER MOLECULE; T-CELL DEVELOPMENT; C-MYC; GENE-EXPRESSION; LACKING JAK3; STEM-CELLS; INDUCTION; BCL-2 AB The common cytokine receptor gamma chain (gamma(c)) is an essential receptor component for IL-2, IL-4, IL-7, IL-9 and IL-15, and thereby gamma(c)-deficient mice exhibit impaired T cell and B cell development. The Janus family tyrosine kinase 3 (Jak3) is known to be associated with gamma(c), and the reported phenotypes of gamma(c)-deficient (gamma(c)(-)) and Jak3-deficient (Jak3(-)) mice are similar, indicating that Jak3 is an essential transducer of gamma(c)-dependent signals. Nevertheless, certain differences have been suggested related to the range of actions of gamma(c) and Jak3, To clarify whether gamma(c)-dependent cytokines can partially transduce their signals without Jak3, we compared lymphocyte development in gamma(c)(-), Jak3(-), and gamma(c) and Jak3 double-deficient (gamma(c)(-)Jak3(-)) mice in the same genetic background. With the exception that T and B cells in Jak3- mice express high levels of gamma(c), the defects in thymocyte and peripheral T cell and B cell development are indistinguishable among gamma(c)(-), Jak3(-) and gamma(c)(-)Jak3(-) mice. Interestingly, although Bcl-2 induction was previously suggested to be Jak3-independent, IL-7 cannot induce Bcl-2 expression in CD4 single-positive (SP) thymocytes in either gamma(c)(-) or Jak3(-) mice nor can IL-7 rescue CD4 SP thymocytes from dexamethasone-induced cell death in gamma(c)(-) or Jak3(-) mice. These results indicate that Jak3 is absolutely essential for gamma(c)-dependent T cell and B cell development, and for gamma(c)-dependent prevention of thymocyte apoptosis. C1 Chiba Univ, Sch Med, Dept Internal Med 2, Chuo Ku, Chiba 2608670, Japan. Chiba Univ, Grad Sch Med, Dept Mol Genet, Chiba 2608670, Japan. NHLBI, Lab Mol Immunol, Bethesda, MD 20892 USA. RP Nakajima, H (reprint author), Chiba Univ, Sch Med, Dept Internal Med 2, Chuo Ku, 1-8-1 Inohana, Chiba 2608670, Japan. RI Saito, Takashi/C-9684-2009 OI Saito, Takashi/0000-0001-9495-3547 NR 65 TC 73 Z9 74 U1 0 U2 3 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0953-8178 J9 INT IMMUNOL JI Int. Immunol. PD FEB PY 2000 VL 12 IS 2 BP 123 EP 132 DI 10.1093/intimm/12.2.123 PG 10 WC Immunology SC Immunology GA 284XZ UT WOS:000085359300001 PM 10653847 ER PT J AU Wallin, KL van Doornum, GJJ Andersson-Ellstrom, A Kallings, I Wiklund, F Hallmans, G Schiller, J Dillner, J AF Wallin, KL van Doornum, GJJ Andersson-Ellstrom, A Kallings, I Wiklund, F Hallmans, G Schiller, J Dillner, J TI Seroepidemiology of human papillomavirus type 73: A sexually transmitted low-risk virus SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article ID SQUAMOUS-CELL CARCINOMA; SERUM ANTIBODIES; HPV DNA; WORLDWIDE PERSPECTIVE; ESOPHAGEAL CANCER; NATURAL-HISTORY; INFECTION; WOMEN; ASSOCIATION; CAPSIDS AB Human papillomavirus type 73 (HPV 73) has been detected in some invasive cervical cancers and has been cloned from a squamous-cell carcinoma of the esophagus, but the epidemiology of this infection and its associated risk of cancer is unknown. We investigated the seroepidemiology of this virus using virus-like particles. The IgG response to HPV 73 appeared to be HPV type-specific, since a comparison of HPV 73 antibody levels before and after infection with HPV 6, 11, 16, 18 or 33 found no evidence of cross-induction of HPV 73 antibodies and since there was little correlation between the antibody levels to HPV 73 and the other 5 investigated HPV types. In both a cross-sectional serosurvey that included 274 women and a 7-year follow-up study that enrolled 98 women, HPV 73 seropositivity was found to be strongly dependent on the number of lifetime sexual partners [OR for > 4 vs. 0 to I partners: 6.0 (95%CI: 1.4-53.6) and 7.9 (95% CI: 2.8-28.3), respectively]. Finally, the risk for HPV 73 seropositive women to develop CIN was investigated in a prospective study nested in a cohort of 15,234 Swedish women. The population-based HPV 73 seroprevalence in Sweden was 14%. No excess risk for CIN was found (OR: 0.77). We conclude that HPV 73 is a mainly sexually transmitted, probably mostly transient, infection that does not confer any measurably increased risk for CIN development. (C) 2000 Wiley-Liss, Inc. C1 Karolinska Inst, Lab Tumor Virus Epidemiol, Ctr Microbiol & Tumor Biol, S-17177 Stockholm, Sweden. Municipal Hlth Serv Amsterdam, Dept Publ Hlth, Amsterdam, Netherlands. Ctr Publ Hlth Res, Karlstad, Sweden. Dist Hlth Ctr Gripen, Karlstad, Sweden. Swedish Inst Infect Dis Control, Dept Bacteriol, Stockholm, Sweden. Umea Univ, Dept Oncol, S-90187 Umea, Sweden. Umea Univ, Med Bank, Dept Publ Hlth & Clin Med, S-90187 Umea, Sweden. NCI, Cellular Oncol Lab, Bethesda, MD 20892 USA. Univ Tampere, Sch Publ Hlth, FIN-33101 Tampere, Finland. RP Wallin, KL (reprint author), Karolinska Inst, Lab Tumor Virus Epidemiol, Ctr Microbiol & Tumor Biol, Box 280, S-17177 Stockholm, Sweden. NR 29 TC 7 Z9 8 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0020-7136 J9 INT J CANCER JI Int. J. Cancer PD FEB 1 PY 2000 VL 85 IS 3 BP 353 EP 357 DI 10.1002/(SICI)1097-0215(20000201)85:3<353::AID-IJC10>3.0.CO;2-M PG 5 WC Oncology SC Oncology GA 271HP UT WOS:000084588400010 PM 10652426 ER PT J AU Perna, AM Bonura, F Vitale, F Viviano, E Di Benedetto, MA Ajello, F Villafrate, MR Prestileo, T Mancuso, S Goedert, JJ Romano, N AF Perna, AM Bonura, F Vitale, F Viviano, E Di Benedetto, MA Ajello, F Villafrate, MR Prestileo, T Mancuso, S Goedert, JJ Romano, N TI Antibodies to human herpes virus type 8 (HHV8) in general population and in individuals at risk for sexually transmitted diseases in Western Sicily SO INTERNATIONAL JOURNAL OF EPIDEMIOLOGY LA English DT Article DE human herpes virus 8; antibodies; epidemiology; human immunodeficiency virus; risk groups; sexually transmitted diseases ID SARCOMA-ASSOCIATED HERPESVIRUS; KAPOSIS-SARCOMA; DNA-SEQUENCES; AIDS; INFECTION; HUMAN-HERPESVIRUS-8 AB Background Human herpes virus 8 (HHV8) appears to be the primary aetiologic agent of Kaposi sarcoma (KS). To study the distribution of HHV8, a seroepidemiological study was carried out in western Sicily, where a high incidence rate of classical KS is well documented. Methods A total of 970 sera of healthy human immunodeficiency virus (HIV) negative individuals of general population (1-70 years old) and 742 sera of individuals in different risk groups for HIV infection were evaluated by means of an indirect immunofluorescence assay able to detect antibodies to lytic and latent HHV8 antigens. Results Crude seroprevalence to HHV8 antigens was 11.5% in the general population, and it increased significantly with age from 6% under age 16 to 22% after age 50. Significantly higher HHV8 seroprevalence rates were detected among HIV positive and negative homosexual men (62% and 22%, respectively), men who had sex with prostitutes (40% and 29%, respectively); female prostitutes (42% and 30%, respectively), and clients at a sexually transmitted disease clinic (male: 60% and 33%, respectively, female: 63% and 43%, respectively). In contrast, heterosexual intravenous drug users had seroprevalence rates comparable to those found in the general population. Conclusions The results suggest that HHV8 infection is widespread in Western Sicily. The high seroprevalence in individuals with high risk sexual activity point to the role of sexual behaviour in the transmission of the infection in adults, whereas the detection of antibodies in younger population (under 16 years old) is suggestive of a non-sexual route of transmission, probably occurring during childhood by close personal contact. C1 Univ Palermo, Policlin, Dipartimento Igiene & Microbiol, Serv Riferimento Reg Diagnost Sierol & Virol AIDS, I-90127 Palermo, Italy. Presidio Osped Casa Sole, Azienda Unita Sanit Locale 6, Palermo, Italy. Univ Palermo, Policlin, Serv Riferimento Reg Diagnosi Clin AIDS & Sindrom, I-90133 Palermo, Italy. NCI, Viral Epidemiol Branch, NIH, Bethesda, MD 20892 USA. RP Romano, N (reprint author), Univ Palermo, Policlin, Dipartimento Igiene & Microbiol, Serv Riferimento Reg Diagnost Sierol & Virol AIDS, Via Vespro 133, I-90127 Palermo, Italy. OI mancuso, salvatrice/0000-0002-7926-090X NR 19 TC 56 Z9 61 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0300-5771 J9 INT J EPIDEMIOL JI Int. J. Epidemiol. PD FEB PY 2000 VL 29 IS 1 BP 175 EP 179 DI 10.1093/ije/29.1.175 PG 5 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 295QB UT WOS:000085978400024 PM 10750620 ER PT J AU Zimonjic, DB Keck-Waggoner, CL Yuan, BZ Kraus, MH Popescu, NC AF Zimonjic, DB Keck-Waggoner, CL Yuan, BZ Kraus, MH Popescu, NC TI Profile of genetic alterations and tumorigenicity of human breast cancer cells SO INTERNATIONAL JOURNAL OF ONCOLOGY LA English DT Article DE breast cancer; proto-oncogenes; gene amplification; tumor suppressor genes; deletion; translocation; gene expression ID COMPARATIVE GENOMIC HYBRIDIZATION; CLONAL CHROMOSOME-ABNORMALITIES; AMPLIFIED DNA-SEQUENCES; C-MYC; SOLID TUMORS; COPY NUMBER; AMPLIFICATION; CARCINOMAS; REGION; ONCOGENE AB The profile of genetic alterations in four breast carcinoma cell lines, SK-BR-3, BT-474, MDA-MB361 and ZR-75-1 was examined by comparative genomic hybridization, G-band karyotyping, reverse chromosome painting and fluorescence in situ hybridization of single-copy genes. These lines are aneuploid with complex structural rearrangements and have DNA copy-number imbalances involving multiple sites that include amplification of ERBB-2 and MYC protooncogenes which are implicated in breast cancer pathogenesis. A novel site of high level amplification was mapped on chromosome 15. All lines were tumorigenic in nude mice, however, the latency and the incidence of tumor formation varied; SK-BR-3 and MDA-MB361 produced tumors in a shorter time and had a higher total number of genomic imbalances compared to BT-474 and ZR-75-1 cells. Tumor cell behavior in vivo was not reflected by the rate of in vitro cell proliferation. Underrepresentation on the long arm of chromosome 18 was the sole alteration that correlated with an increased tumorigenicity. Chromosome 18q is rich in tumor suppressor genes and its loss is prevalent in. primary node-positive breast tumors. Cell lines with monoclonal populations preserve the genetic characteristics of the primary tumor and their use may facilitate the detection of specific alterations associated with breast cancer progression. C1 NCI, Expt Carcinogenesis Lab, Bethesda, MD 20892 USA. European Inst Oncol, Dept Expt Oncol, Milan, Italy. RP Popescu, NC (reprint author), NCI, Expt Carcinogenesis Lab, Bldg 37,Room 3C05,37 Convent Dr,MSC 4255, Bethesda, MD 20892 USA. NR 55 TC 17 Z9 17 U1 0 U2 5 PU PROFESSOR D A SPANDIDOS PI ATHENS PA 1, S MERKOURI ST, EDITORIAL OFFICE,, ATHENS 116 35, GREECE SN 1019-6439 J9 INT J ONCOL JI Int. J. Oncol. PD FEB PY 2000 VL 16 IS 2 BP 221 EP 230 PG 10 WC Oncology SC Oncology GA 275FL UT WOS:000084808700001 PM 10639563 ER PT J AU Cartee, L Vrana, JA Wang, ZL Park, JS Birrer, M Fisher, PB Grant, S Dent, P AF Cartee, L Vrana, JA Wang, ZL Park, JS Birrer, M Fisher, PB Grant, S Dent, P TI Inhibition of the mitogen activated protein kinase pathway potentiates radiation-induced cell killing via cell cycle arrest at the G(2)/M transition and independently of increased signaling by the JNK/c-Jun pathway SO INTERNATIONAL JOURNAL OF ONCOLOGY LA English DT Article DE MAPK; JNK; c-Jun; p21(Cip-1/WAF1); ionizing radiation; proliferation; cell death; apoptosis ID MYELOID-LEUKEMIA CELLS; N-TERMINAL KINASE; INDUCED APOPTOSIS; INDUCED PROLIFERATION; RAT HEPATOCYTES; STRESS; CERAMIDE; SURVIVAL; CASCADE AB The ability of low dose ionizing radiation (2 Gy) to modulate the activities of the mitogen activated protein kinase (MAPK) and c-Jun NH2-terminal kinase (JNK1) cascades in human monocytic leukemia (U937/pREP4) cells and in cells over-expressing dominant negative c-Jun (TAM67) (U937/ TAM67) was investigated. Radiation exposure caused prolonged (similar to 1 h) MAPK activations in U937 cells. In contrast, low dose irradiation weakly modulated JNK1 activity in these cells. Inhibition of the MAPK pathway by use of the specific MEK1/2 inhibitor (10 mu M PD98059) in both U937/pREP4 and U937/TAM67 cells prior to radiation exposure permitted strong prolonged radiation-induced activations of JNK1. Expression of TAM67 decreased the ability of radiation to cause apoptosis compared to control transfected cells. However, combined MEK1/2 inhibition and radiation exposure in both cell types caused a large decrease in suspension culture growth and a large increase in apoptosis, when compared to either treatment alone. Reduced proliferation after combined irradiation and PD98059 treatment in both cell types correlated with prolonged cell cycle arrest in G(2)/M phase. Prolonged growth arrest was abolished when MEK1/2 inhibitor was removed 6 h following irradiation, which was associated with a reduction in apoptosis. The ability of MEK1/2 inhibition to cause prolonged G(2)/M growth arrest was reduced in U937 cells stably transfected with a p21(Cip-1/WAF1) antisense construct of radiation-induced apoptosis and a reduced ability of MEK1/2 inhibition to potentiate apoptosis. Collectively our data demonstrate that inhibition of MEK1/2 function increases the radiation sensitivity of U937 cells, independently of c-Jun function, and decreases the ability of these cells to recover from the radiation-induced G(2)/M cell cycle checkpoint arrest. In addition, our data also demonstrate that the ability of MEK1/2 inhibition to potentiate radiation-induced cell death in U937 cells in part requires an ability of cells to express low levels of p21(Cip-1/WAF1). C1 Virginia Commonwealth Univ, Med Coll Virginia, Dept Radiat Oncol, Richmond, VA 23298 USA. Virginia Commonwealth Univ, Med Coll Virginia, Dept Med, Richmond, VA 23298 USA. Virginia Commonwealth Univ, Med Coll Virginia, Dept Pharmacol & Toxicol, Richmond, VA 23298 USA. NCI, Biomarkers Branch, Div Clin Sci, Rockville, MD 20850 USA. Columbia Univ Coll Phys & Surg, New York, NY 10032 USA. RP Dent, P (reprint author), Virginia Commonwealth Univ, Med Coll Virginia, Dept Radiat Oncol, 401 Coll St, Richmond, VA 23298 USA. FU NCI NIH HHS [P01-CA72955, R01-CA77141, R01-CA63753] NR 35 TC 24 Z9 24 U1 0 U2 1 PU PROFESSOR D A SPANDIDOS PI ATHENS PA 1, S MERKOURI ST, EDITORIAL OFFICE,, ATHENS 116 35, GREECE SN 1019-6439 J9 INT J ONCOL JI Int. J. Oncol. PD FEB PY 2000 VL 16 IS 2 BP 413 EP 422 PG 10 WC Oncology SC Oncology GA 275FL UT WOS:000084808700024 PM 10639586 ER PT J AU Pietrini, P Furey, ML Guazzelli, M AF Pietrini, P Furey, ML Guazzelli, M TI The neural basis of cognitive decline: Functional brain studies in humans SO INTERNATIONAL JOURNAL OF PSYCHOPHYSIOLOGY LA English DT Meeting Abstract C1 Univ Pisa, Dept Human & Environm Sci, Pisa, Italy. NIA, Neurosci Lab, Bethesda, MD 20892 USA. NIMH, Lab Brain & Cognit, NIH, Bethesda, MD 20892 USA. Univ Pisa, Dept Psychiat Neurobiol Pharmacol & Biotechnol, Pisa, Italy. NR 0 TC 0 Z9 0 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0167-8760 J9 INT J PSYCHOPHYSIOL JI Int. J. Psychophysiol. PD FEB PY 2000 VL 35 IS 1 BP 12 EP 12 PG 1 WC Psychology, Biological; Neurosciences; Physiology; Psychology; Psychology, Experimental SC Psychology; Neurosciences & Neurology; Physiology GA 273UY UT WOS:000084727300027 ER PT J AU Duncan, MK Cvekl, A Li, X Piatigorsky, J AF Duncan, MK Cvekl, A Li, X Piatigorsky, J TI Truncated forms of Pax-6 disrupt lens morphology in transgenic mice SO INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE LA English DT Article ID MISSENSE MUTATIONS; CRYSTALLIN GENE; PAIRED DOMAIN; SMALL EYE; ANIRIDIA; EXPRESSION; CATARACT; MOUSE; TRANSCRIPTION; ANTERIOR AB PURPOSE. Extensive Literature shows that Pax-6 is critical fur lens development and that Pax-6 mutations can result in aniridia in humans. In addition, it has been reported that truncated Pax-6 molecules can act as dominant-negative repressors of wild-type Pax-6 activity in cultured cells. This study was designed to determine whether Pax-6 molecules without either the activation domain (AD) or the homeodomain (HD) and the AD can function as dominant-negative repressors in vivo and alter the phenotype of the lens. METHODS. Transgenic mice were created harboring the alpha A-crystallin promoter linked to a cDNA encoding either a truncated Pax-6 without the C terminus (paired domain [PD] + homeodomain) or Pax-6 consisting of only the PD. The phenotype of the resultant animals was investigated by light and electron microscopy as well as atomic absorption spectroscopy. RESULTS. Two lines of PD + HD mice and three lines of PD mice were generated,all of which exhibit posterior nuclear and/or cortical cataracts of variable severity. The lenses from mice transgenic for either Pax-6 truncation are smaller and more hydrated than normal. Morphologically, the mice expressing the PD + HD of Pax-6 have swollen lens fibers with attenuated ball-and-socket junctions. In contrast, the lenses from mice overexpressing the PD of Pax-6 have posterior nuclear cataracts composed of cell debris, whereas the remaining fiber cells appear generally normal. CONCLUSIONS. The presence of truncated Pax-6 protein in the lens is sufficient to induce cataract in a wild-type genetic background. The simplest explanation for this phenomenon is a dominant-negative effect; however, a number of other possible mechanisms are presented. C1 Univ Delaware, Dept Biol Sci, Newark, DE 19716 USA. Albert Einstein Coll Med, Dept Ophthalmol, Bronx, NY 10467 USA. Albert Einstein Coll Med, Dept Mol Genet, Bronx, NY 10467 USA. NEI, Mol & Dev Biol Lab, Bethesda, MD 20892 USA. RP Duncan, MK (reprint author), Univ Delaware, Dept Biol Sci, 331 Wolf Hall, Newark, DE 19716 USA. RI Cvekl, Ales/B-2427-2013 FU NCRR NIH HHS [RR11820]; NEI NIH HHS [1Z01EY00126-17, R01 EY12221-01] NR 50 TC 40 Z9 43 U1 0 U2 2 PU ASSOC RESEARCH VISION OPHTHALMOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0146-0404 J9 INVEST OPHTH VIS SCI JI Invest. Ophthalmol. Vis. Sci. PD FEB PY 2000 VL 41 IS 2 BP 464 EP 473 PG 10 WC Ophthalmology SC Ophthalmology GA 282EP UT WOS:000085204100019 PM 10670477 ER PT J AU Cheng, QF Lichtstein, D Russell, P Zigler, JS AF Cheng, QF Lichtstein, D Russell, P Zigler, JS TI Use of a lipophilic cation to monitor electrical membrane potential in the intact rat lens SO INVESTIGATIVE OPHTHALMOLOGY & VISUAL SCIENCE LA English DT Article ID GLIOMA-CELLS; METABOLISM; MONENSIN; SYNAPTOSOMES; TRANSPORT; CATARACTS; ACID; NA+ AB PURPOSE. Tetraphenylphosphonium (TPP+) is a permeant lipophilic cation that accumulates in cultured cells and tissues as a function of the electrical membrane potential across the plasma membrane. This study was undertaken to determine whether TPP+ can be used for assessing membrane potential in intact lenses in organ culture. METHODS. Rat lenses were cultured in media containing 10 mu M TPP+ and a tracer level of H-3-TPP+ for various times. H-3-TPP+ levels in whole lenses or dissected portions of lenses were determined by liquid scintillation counting. Ionophores, transport inhibitors, and neurotransmitters were also added to investigate their effects on TPP+ uptake. RESULTS. Incubation of lenses in low-K+ balanced salt solution and TC-199 medium, containing physiological concentrations of Na+ and K+, led to a biphasic accumulation of TPP+ in the lens that approached equilibrium by 12 to 16 hours of culture, the TPP+ equilibrated within 1 hour in the epithelium but penetrated more slowly into the fiber mass. The steady state level of TPP+ accumulation in the lens was depressed by 90% when the lenses were cultured in a medium containing high K+. The calculated membrane potential for normal rat lens in TC-199 was -75 +/- 3 mV. Monensin (1 mu M) and nigericin (1 mu M). Na+H+ and K+H+ exchangers respectively, as well as the protonophore carbonylcyanide-m-chlorophenylhydrazone (CCCP, 10 mu M) and the calcium ionophore A23187 (10 mu M), abolished TPP+ accumulation and caused cloudiness of the lenses. The neurotransmitter acetylcholine at 50 mu M decreased TPP+ accumulation in the lens, but this effect could be prevented by simultaneous application of 1 mM atropine. CONCLUSIONS. TPP+ accumulation can be used as an indicator of changes in membrane potential in intact lenses, but because of the long time required to reach steady state, its utility is limited., The slow accumulation of TPP+ and its slow efflux from the lens under conditions known to depolarize membranes are consistent with a diffusion barrier in the deep cortex and nucleus of the lens. C1 NEI, Lab Mechanisms Ocular Dis, NIH, Bethesda, MD 20892 USA. Hebrew Univ Jerusalem, Hadassah Med Sch, Dept Physiol, IL-91010 Jerusalem, Israel. RP Zigler, JS (reprint author), NEI, Lab Mechanisms Ocular Dis, NIH, 6 Ctr Dr,MSC 2735, Bethesda, MD 20892 USA. NR 26 TC 5 Z9 5 U1 2 U2 2 PU ASSOC RESEARCH VISION OPHTHALMOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0146-0404 J9 INVEST OPHTH VIS SCI JI Invest. Ophthalmol. Vis. Sci. PD FEB PY 2000 VL 41 IS 2 BP 482 EP 487 PG 6 WC Ophthalmology SC Ophthalmology GA 282EP UT WOS:000085204100021 PM 10670479 ER PT J AU Choi, YH Lee, WH Park, KY Zhang, LJ AF Choi, YH Lee, WH Park, KY Zhang, LJ TI p53-independent induction of p21 (WAF1/CIP1), reduction of cyclin B1 and G2/M arrest by the isoflavone genistein in human prostate carcinoma cells SO JAPANESE JOURNAL OF CANCER RESEARCH LA English DT Article DE genistein; cell cycle; G2/M arrest; p21 ID KINASE INHIBITOR GENISTEIN; DNA STRAND BREAKAGE; LEUKEMIA-CELLS; DEPENDENT KINASES; MAMMALIAN FIBROBLASTS; SIGNAL TRANSDUCTION; MELANOMA-CELLS; G(2)/M ARREST; CANCER-CELLS; G(1) ARREST AB Genistein. a natural isoflavonoid phytoestrogen, is a strong inhibitor of protein tyrosine kinase and DNA topoisomerase II activities. Genistein has been shown to have anticancer proliferation, differentiation and chemopreventive effects. In the present study, we have addressed the mechanism of action by which genistein suppressed the proliferation of p53-null human prostate carcinoma cells. Genistein significantly inhibited the cell growth, which effect was reversible, and induced dendrite-like structure, The inhibitory effects of genistein on cell growth proliferation were associated with a G2/M arrest in cell cycle progression concomitant with a marked inhibition of cyclin B1 and an induction of Cdk inhibitor p21 (WAFI/CIP1) in a p53-independent manner. Following genistein treatment of cells, an increased binding of p21 with Cdk2 and Cdc2 paralleled a significant decrease in Cdc2 and Cdk2 kinase activity with no change in Cdk2 and Cdc2 expression. Genistein also induced the activation of a p21 promoter reporter construct, utilizing a sequence distinct from the p53-binding site, Analysis of deletion constructs of the p21 promoter indicated that the response to genistein could be localized to the 300 base pairs proximal to the transcription start site. These data suggest that genistein may exert a strong anticarcinogenic effect, and that this effect possibly involves an induction of p21, which inhibits the threshold kinase activities of Cdks and associated cyclins, leading to a G2/M arrest in the cell cycle progression. C1 Pusan Natl Univ, Pusan Canc Res Ctr, Dept Food Sci & Nutr, Pusan 609735, South Korea. Pusan Natl Univ, Pusan Canc Res Ctr, Coll Nat Sci, Dept Biol, Pusan 609735, South Korea. NCI, Pediat Oncol Branch, NIH, Bethesda, MD 20892 USA. RP Park, KY (reprint author), Pusan Natl Univ, Pusan Canc Res Ctr, Dept Food Sci & Nutr, Pusan 609735, South Korea. NR 56 TC 123 Z9 127 U1 0 U2 2 PU BUSINESS CENTER ACADEMIC SOCIETIES JAPAN PI TOKYO PA 5-16-9 HONKOMAGOME, BUNKYO-KU, TOKYO, 113-8633, JAPAN SN 0910-5050 J9 JPN J CANCER RES JI Jpn. J. Cancer Res. PD FEB PY 2000 VL 91 IS 2 BP 164 EP 173 PG 10 WC Oncology SC Oncology GA 286PP UT WOS:000085454600004 PM 10761703 ER PT J AU Epstein, FH Arai, AE AF Epstein, FH Arai, AE TI Optimization of fast cardiac imaging using an echo-train readout SO JMRI-JOURNAL OF MAGNETIC RESONANCE IMAGING LA English DT Article DE cardiac imaging; echoplanar imaging; optimization ID SINGLE BREATH-HOLD; STEADY-STATE; HUMAN-HEART; PLANAR; MAGNETIZATION; CONTRAST; MOTION; MRI; EPI AB Fast gradient-echo sequences that use an echo-train readout are becoming more widely used, particularly for imaging the heart. An important issue for these sequences involves determining the optimal duration for the echo-train readout, In normal volunteer scans and theoretically the echo-train readout duration was varied from 2.4 to 32.8 msec, Myocardial signal-to-noise ratio [SNR], myocardiumtag signal difference-to-noise ratio (SDNR), now artifact-to-noise ratio (FNR), and geometric distortion were measured and/or calculated, Our results showed that to obtain high SNR, SDNR, and data acquisition efficiency while minimizing FNR and geometric distortion, the readout duration should be 10-15 msec at 1.5 T. Published 2000 Wiley-Liss, Inc.(dagger) C1 NHLBI, Cardiac Energet Lab, NIH, Bethesda, MD 20892 USA. RP Epstein, FH (reprint author), NHLBI, Cardiac Energet Lab, NIH, Bldg 10,Room B1D416,10 Ctr Dr,MSC 1061, Bethesda, MD 20892 USA. NR 26 TC 18 Z9 18 U1 0 U2 0 PU JOHN WILEY & SONS INC PI NEW YORK PA 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 1053-1807 J9 JMRI-J MAGN RESON IM JI JMRI-J. Magn. Reson. Imaging PD FEB PY 2000 VL 11 IS 2 BP 75 EP 80 DI 10.1002/(SICI)1522-2586(200002)11:2<75::AID-JMRI1>3.0.CO;2-P PG 6 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 297JA UT WOS:000086078100001 PM 10713937 ER PT J AU Hertle, RW Zhu, XS AF Hertle, RW Zhu, XS TI Oculographic and clinical characterization of thirty-seven children with anomalous head postures, nystagmus, and strabismus: The basis of a clinical algorithm SO JOURNAL OF AAPOS LA English DT Article ID MANIFEST LATENT NYSTAGMUS AB Background and Purpose: We studied children with nystagmus who also had anomalous head postures and strabismus to determine the etiology of the conditions and present a diagnostic clinical algorithm. Methods: The patients for this study were among the 560 patients evaluated in the ocular motor neurophysiology laboratory between the years 1991 and 1997, Clinical characteristics, infrared oculography data, and medical and surgical treatments were entered into a database for analysis. Oculography was performed on all patients according to a standard protocol, and data were stored and analyzed off-line. Etiology of anomalous head posture was determined with both clinical and oculography information. Results: Thirty-seven children are the subjects of this report. The etiology of anomalous head posture was a "gaze null" due to congenital nystagmus in 23 (62%) patients, an "adduction null" due to manifest latent nystagmus in 12 (32%) patients, spasmus nutans in 1 (3%) patient, and strabismus in 1 (3%) patient. The patients' ages ranged from 9 months to 12 years and averaged 4.4 years. Sixty-nine percent were male patients. Nineteen (63%) of 30 patients had abnormal recognition (linear optotype) acuity in at least 1 eye on monocular cover; the recognition remained abnormal in 5 (17%) of 30 patients under binocular conditions. Thirty percent of patients had amblyopia, 16% had some structural disease of the eyes, 22% had some systemic syndrome or abnormality, 57% had a significant refractive error, and 27% had some ability to fuse. Conclusions: The major etiology for anomalous head posture in these patients was to adopt a gaze null due to congenital nystagmus (62% of patients) regardless of the direction of their anomalous head posture or type of strabismus. Moving the fixing eye as the first step for the anomalous head posture, combined with moving the nonfixing eye for the resulting strabismus may help treat these patients. C1 NEI, Sensorimotor Res Lab, NIH, Bethesda, MD 20892 USA. Univ Penn, Childrens Hosp Philadelphia, Philadelphia, PA 19104 USA. RP Hertle, RW (reprint author), NEI, Sensorimotor Res Lab, NIH, Bldg 49 Room 2A50, Bethesda, MD 20892 USA. NR 28 TC 18 Z9 22 U1 0 U2 1 PU MOSBY-YEAR BOOK INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 1091-8531 J9 J AAPOS JI J. AAPOS PD FEB PY 2000 VL 4 IS 1 BP 25 EP 32 DI 10.1016/S1091-8531(00)90008-9 PG 8 WC Ophthalmology; Pediatrics SC Ophthalmology; Pediatrics GA 286PW UT WOS:000085455200005 PM 10675868 ER PT J AU Hertle, RW Dell'Osso, LF AF Hertle, RW Dell'Osso, LF TI Congenital nystagmus: In search of simplicity on the other side of complexity SO JOURNAL OF AAPOS LA English DT Letter C1 NEI, Sensorimotor Res Lab, Bethesda, MD 20892 USA. NIH, Bethesda, MD 20892 USA. Case Western Reserve Univ, Dept Neurol, Cleveland, OH 44106 USA. Case Western Reserve Univ, Vet Affairs Med Ctr, Cleveland, OH 44106 USA. RP Hertle, RW (reprint author), NEI, Sensorimotor Res Lab, Bldg 10, Bethesda, MD 20892 USA. NR 2 TC 0 Z9 0 U1 0 U2 1 PU MOSBY-YEAR BOOK INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 1091-8531 J9 J AAPOS JI J. AAPOS PD FEB PY 2000 VL 4 IS 1 BP 62 EP 62 DI 10.1016/S1091-8531(00)90016-8 PG 1 WC Ophthalmology; Pediatrics SC Ophthalmology; Pediatrics GA 286PW UT WOS:000085455200013 PM 10675876 ER PT J AU Scheidt, P Overpeck, M Wyatt, W Ross, J Aszmann, A AF Scheidt, P Overpeck, M Wyatt, W Ross, J Aszmann, A TI Health perceptions and symptoms of young adolescents: An international study SO JOURNAL OF ADOLESCENT HEALTH LA English DT Meeting Abstract C1 NICHHD, Bethesda, MD 20892 USA. Childrens Natl Med Ctr, Washington, DC 20010 USA. Macro Int, Calverton, MD USA. Natl Inst Child Hlth, Budapest, Hungary. NR 0 TC 0 Z9 0 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 1054-139X J9 J ADOLESCENT HEALTH JI J. Adolesc. Health PD FEB PY 2000 VL 26 IS 2 BP 123 EP 123 PG 1 WC Psychology, Developmental; Public, Environmental & Occupational Health; Pediatrics SC Psychology; Public, Environmental & Occupational Health; Pediatrics GA 285NZ UT WOS:000085396300081 ER PT J AU Cheng, TL Wright, JL Fields, CB Brenner, RA O'Donnell, R Schwarz, D Scheidt, PC AF Cheng, TL Wright, JL Fields, CB Brenner, RA O'Donnell, R Schwarz, D Scheidt, PC TI Violent injuries among adolescents: Declining morbidity and mortality in an urban population SO JOURNAL OF ADOLESCENT HEALTH LA English DT Meeting Abstract C1 Childrens Natl Med Ctr, MPH & DC Child Adolescent Injury Res Network, Washington, DC 20010 USA. George Washington Univ, Washington, DC USA. NICHHD, Bethesda, MD 20892 USA. Childrens Hosp Philadelphia, Philadelphia, PA 19104 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 1054-139X J9 J ADOLESCENT HEALTH JI J. Adolesc. Health PD FEB PY 2000 VL 26 IS 2 BP 146 EP 146 PG 1 WC Psychology, Developmental; Public, Environmental & Occupational Health; Pediatrics SC Psychology; Public, Environmental & Occupational Health; Pediatrics GA 285NZ UT WOS:000085396300127 ER PT J AU Magone, MT Whitcup, SM Fukushima, A Chan, CC Silver, PB Rizzo, LV AF Magone, MT Whitcup, SM Fukushima, A Chan, CC Silver, PB Rizzo, LV TI The role of IL-12 in the induction of late-phase cellular infiltration in a murine model of allergic conjunctivitis SO JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY LA English DT Article DE IL-12; IFN-gamma; allergic conjunctivitis; cytokine-deficient mice; IL-4 ID INDUCED AIRWAY HYPERRESPONSIVENESS; EXPERIMENTAL AUTOIMMUNE UVEITIS; INDUCED EOSINOPHIL INFILTRATION; COLONY-STIMULATING FACTOR; IFN-GAMMA; DEFICIENT MICE; INTERLEUKIN-4-DEFICIENT MICE; INTERFERON-GAMMA; IL-12-DEFICIENT MICE; OCULAR ALLERGY AB Background: The applied murine model of allergic conjunctivitis mimics human disease, and an immediate hypersensitivity reaction (LHR) and a late-phase cellular reaction typically develop in sensitized mice after topical challenge with the allergen. Objective: We investigated the role of IL-4, IFN-gamma, and IL-12 in the early and late phases of ocular allergy with use of cytokine knockout (KO) mice and neutralizing antibodies. Methods: Ragweed-sensitized wild-type or IL-4KO, IL-12KO, IFN-gamma KO, anti-IL-12 mAb-treated, recombinant murine IL-12-treated, and anti-IFN-gamma mAb-treated mice were challenged with the allergen 10 days after the immunization, IHR, cellular infiltration, lymphoproliferative response, and cytokine production from draining lymph nodes were recorded and compared among groups. Results: We show that IL-12KO mice and anti-IL-12 antibody-treated wild-type animals failed to have a cellular infiltration into the conjunctiva. Treatment with recombinant murine IL-12 also reduced the number of infiltrating PMNs but increased the percentage of mononuclear cells in the conjunctiva compared with controls, IFN-gamma KO mice had a significantly stronger MR and prolonged infiltration into the conjunctiva after challenge with ragweed than controls. Conclusion: Our data suggest that the presence of IL-12, although better known as a T(H)1-inducing cytokine, is important for the development and the regulation of the late-phase pathologic features in ocular allergy. Furthermore, IFN-gamma is a limiting factor in the late phase of allergy and thus may be important in preventing chronic allergic disease. C1 NEI, NIH, Bethesda, MD 20892 USA. Kochi Med Sch, Kochi, Japan. Univ Sao Paulo, Sao Paulo, Brazil. RP Magone, MT (reprint author), NEI, NIH, Bldg 10,Rm 10N208, Bethesda, MD 20892 USA. RI Rizzo, Luiz Vicente/B-4458-2009 NR 63 TC 41 Z9 41 U1 2 U2 3 PU MOSBY-YEAR BOOK INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0091-6749 J9 J ALLERGY CLIN IMMUN JI J. Allergy Clin. Immunol. PD FEB PY 2000 VL 105 IS 2 BP 299 EP 308 DI 10.1016/S0091-6749(00)90080-0 PN 1 PG 10 WC Allergy; Immunology SC Allergy; Immunology GA 285RQ UT WOS:000085402200015 PM 10669851 ER PT J AU Smith, WB AF Smith, WB TI Commentary on Schoenthaler et al.: Vitamin and mineral supplements - Is the methodology sufficient to support the conclusions? SO JOURNAL OF ALTERNATIVE AND COMPLEMENTARY MEDICINE LA English DT Editorial Material ID CONTROLLED TRIAL; INTELLIGENCE; SCHOOLCHILDREN C1 NIAAA, DCPR, NIH, Bethesda, MD 20892 USA. RP Smith, WB (reprint author), NIAAA, DCPR, NIH, 6000 Execut Blvd,Suite 505, Bethesda, MD 20892 USA. NR 10 TC 4 Z9 4 U1 0 U2 0 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1075-5535 J9 J ALTERN COMPLEM MED JI J. Altern. Complement Med. PD FEB PY 2000 VL 6 IS 1 BP 31 EP 35 DI 10.1089/acm.2000.6.31 PG 5 WC Integrative & Complementary Medicine SC Integrative & Complementary Medicine GA 285PN UT WOS:000085397600005 PM 10706233 ER PT J AU Sandkvist, M Keith, JM Bagdasarian, M Howard, SP AF Sandkvist, M Keith, JM Bagdasarian, M Howard, SP TI Two regions of EpsL involved in species-specific protein-protein interactions with EpsE and EpsM of the general secretion pathway in Vibrio cholerae SO JOURNAL OF BACTERIOLOGY LA English DT Article ID GRAM-NEGATIVE BACTERIA; OUTER-MEMBRANE BIOGENESIS; PSEUDOMONAS-AERUGINOSA; ESCHERICHIA-COLI; AEROMONAS-HYDROPHILA; TOXIN SECRETION; EXTRACELLULAR SECRETION; ERWINIA-CHRYSANTHEMI; PREPILIN PEPTIDASE; RNA-POLYMERASE AB Extracellular secretion of proteins via the type II or general secretion pathway in gram-negative bacteria requires the assistance of at least 12 gene products that are thought to form a complex apparatus through which secreted proteins are translocated. Although this apparatus is specifically required only for the outer membrane translocation step during transport across the bacterial cell envelope, it is believed to span both membranes. The EpsE, EpsL, and EpsM proteins of the type II apparatus in Vibrio cholerae are thought to form a trimolecular complex that is required to either control the opening and closing of the secretion pore or to transduce energy to the site of outer membrane translocation. EpsL is likely to play an important role in this relay by interacting with both the cytoplasmic EpsE protein and the cytoplasmic membrane protein EpsM, which is predominantly exposed on the periplasmic side of the membrane. We have now extended this model and mapped the separate regions within EpsL that contain the EpsE and EpsM binding domains. By taking advantage of the species specificity of the type II pathway, we have used chimeric proteins composed of EpsL and its homologue, ExeL, from Aeromonas hydrophila together with either EpsE or its Aeromonas homologue, ExeE, to complement the secretion defect in both epsL and exeL mutant strains, These studies have mapped the species-specific EpsE binding site to the N-terminal cytoplasmic region between residues 57 and 216 of EpsL. In addition, the species-specific EpsM binding site was mapped to the C-terminal half of EpsL by coimmunoprecipitation of EpsM with different EpsL-ExeL chimeras. This site is present in the region between amino acids 216 and 296, which contains the predicted membrane-spanning segment of EpsL. C1 Amer Red Cross, Dept Biochem, Holland Lab, Rockville, MD 20855 USA. NIDR, Vaccine & Therapeut Dev Sect, Oral Infect & Immun Branch, NIH, Bethesda, MD 20892 USA. Michigan State Univ, Dept Microbiol, E Lansing, MI 48824 USA. Univ Regina, Dept Biol, Regina, SK S4S 0A2, Canada. RP Sandkvist, M (reprint author), Amer Red Cross, Dept Biochem, Holland Lab, 15601 Crabbs Branch Way, Rockville, MD 20855 USA. NR 43 TC 41 Z9 44 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 USA SN 0021-9193 J9 J BACTERIOL JI J. Bacteriol. PD FEB PY 2000 VL 182 IS 3 BP 742 EP 748 DI 10.1128/JB.182.3.742-748.2000 PG 7 WC Microbiology SC Microbiology GA 278RP UT WOS:000085003000023 PM 10633109 ER PT J AU Smaili, SS Hsu, YT Youle, RJ Russell, JT AF Smaili, SS Hsu, YT Youle, RJ Russell, JT TI Mitochondria in Ca2+ signaling and apoptosis SO JOURNAL OF BIOENERGETICS AND BIOMEMBRANES LA English DT Review DE Ca2+ signaling; inositol 1,4,5-trisphosphate receptor; mitochondrial Ca2+ uptake; mitochondrial Ca2+ efflux; permeability transition; apoptosis; Bcl-2 family ID PERMEABILITY TRANSITION PORE; CYTOCHROME-C RELEASE; ADENINE-NUCLEOTIDE TRANSLOCASE; PROGRAMMED CELL-DEATH; ENDOPLASMIC-RETICULUM; CALCIUM-UPTAKE; HEART-MITOCHONDRIA; BCL-2 ONCOPROTEIN; ENDOTHELIAL-CELLS; CYCLOSPORINE-A AB Cellular Ca2+ signals are crucial in the control of most physiological processes, cell injury and programmed cell death; mitochondria play a pivotal role in the regulation of such cytosolic Ca2+ ([Ca2+](c)) signals. Mitochondria are endowed with multiple Ca2+ transport mechanisms by which they take up and release Ca2+ across their inner membrane. These transport processes function to regulate local and global [Ca2+](c), thereby regulating a number of Ca2+-sensitive cellular mechanisms. The permeability transition pore (PTP) forms the major Ca2+ efflux pathway from mitochondria. In addition, Ca2+ efflux from the mitochondrial matrix occurs by the reversal of the uniporter and through the inner membrane Na+/Ca2+ exchanger. During cellular Ca2+ overload, mitochondria take up [Ca2+](c), which, in turn, induces opening of PTP, disruption of mitochondrial membrane potential (Delta Psi(m)) and cell death. In apoptosis signaling, collapse of Delta Psi(m) and cytochrome c release from mitochondria occur followed by activation of caspases, DNA fragmentation, and cell death. Translocation of Bar, an apoptotic signaling protein from the cytosol to the mitochondrial membrane, is another step during this apoptosis-signaling pathway. The role of permeability transition in the context of cell death in relation to Bcl-2 family of proteins is discussed. C1 NICHHD, Lab Cellular & Mol Neurophysiol, NIH, Bethesda, MD 20892 USA. Univ Fed Sao Paulo, Dept Farmacol, Sao Paulo, Brazil. NINDS, Biochem Sect, Surg Neurol Branch, NIH, Bethesda, MD 20892 USA. RP Russell, JT (reprint author), NICHHD, Lab Cellular & Mol Neurophysiol, NIH, Bethesda, MD 20892 USA. NR 100 TC 112 Z9 119 U1 0 U2 9 PU KLUWER ACADEMIC/PLENUM PUBL PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013 USA SN 0145-479X J9 J BIOENERG BIOMEMBR JI J. Bioenerg. Biomembr. PD FEB PY 2000 VL 32 IS 1 BP 35 EP 46 DI 10.1023/A:1005508311495 PG 12 WC Biophysics; Cell Biology SC Biophysics; Cell Biology GA 306ZZ UT WOS:000086629900005 PM 11768760 ER PT J AU Herynek, V Bulte, JWM Douglas, T Brooks, RA AF Herynek, V Bulte, JWM Douglas, T Brooks, RA TI Dynamic relaxometry: application to iron uptake by ferritin SO JOURNAL OF BIOLOGICAL INORGANIC CHEMISTRY LA English DT Article DE ferritin; dynamic relaxometry; paramagnetism; antiferromagnetism ID HORSE SPLEEN APOFERRITIN; CORE FORMATION; HUMAN-H; KINETICS AB We introduce dynamic relaxometry as a novel technique for studying biochemical reactions, such as those leading to mineral formation (biomineralization). This technique was applied to follow the time course of iron oxidation and hydrolysis by the protein ferritin. Horse spleen apoferritin was loaded with single additions of 4, 10, 20, 40, and 100 ferrous ions per protein, and with multiple additions of 4, 10, 20, and 100 ferrous ions. The NMR T-2 relaxation time was then measured sequentially and continuously for up to 24 h. At low loading factors of 4-10 Fe atoms/molecule, the iron is rapidly bound and oxidized by the protein on a time scale of approximately 15 s to several minutes. At intermediate loading factors (10-40), rapid initial oxidation was observed, followed by the formation of antiferromagnetic clusters. This process occurred at a much slower rate and continued for up to several hours, but was inhibited at lower pH values. At higher loading factors (40-1000), iron oxidation may occur directly on the core, and this process may continue for up to 24 h following the initial loading. Dynamic relaxometry appears to be a potentially powerful technique that may well have applications beyond the study of iran upake by the ferritin protein. C1 NIH, Lab Diagnost Radiol Res, Bethesda, MD 20892 USA. NINDS, Neuroimaging Branch, NIH, Bethesda, MD 20892 USA. Temple Univ, Dept Chem, Philadelphia, PA 19122 USA. RP Bulte, JWM (reprint author), NIH, Lab Diagnost Radiol Res, Bldg 10,Room B1N256,10 Ctr Dr,MSC 1074, Bethesda, MD 20892 USA. RI Bulte, Jeff/A-3240-2008; Douglas, Trevor/F-2748-2011 OI Bulte, Jeff/0000-0003-1202-1610; NR 22 TC 6 Z9 7 U1 0 U2 3 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0949-8257 J9 J BIOL INORG CHEM JI J. Biol. Inorg. Chem. PD FEB PY 2000 VL 5 IS 1 BP 51 EP 56 PG 6 WC Biochemistry & Molecular Biology; Chemistry, Inorganic & Nuclear SC Biochemistry & Molecular Biology; Chemistry GA 295EZ UT WOS:000085954900006 PM 10766436 ER PT J AU Cowan, JA Ohyama, T Howard, K Rausch, JW Cowan, SML Le Grice, SFJ AF Cowan, JA Ohyama, T Howard, K Rausch, JW Cowan, SML Le Grice, SFJ TI Metal-ion stoichiometry of the HIV-1 RT ribonuclease H domain: evidence for two mutually exclusive sites leads to new mechanistic insights on metal-mediated hydrolysis in nucleic acid biochemistry SO JOURNAL OF BIOLOGICAL INORGANIC CHEMISTRY LA English DT Article DE human immunodeficiency virus type 1 reverse transcriptase ribonuclease H domain; calorimetry; metal binding; mechanism; stoichiometry ID RETROVIRAL REVERSE-TRANSCRIPTASE; DEPENDENT DNA POLYMERASE; CRYSTAL-STRUCTURE; ACTIVE-SITE; ANGSTROM RESOLUTION; POINT MUTATIONS; SARCOMA VIRUS; STRANDED-DNA; RNASE-H; BINDING AB Crystallographic studies of the Mn2+-doped RNase H domain of human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) [1] have revealed two bound Mn2+ separated by approximately 4 Angstrom and surrounded by a cluster of four conserved carboxylates. Escherichia coli RNase H is structurally similar to the RNase H domain of HIV-1 RT, but requires one divalent metal cation for its activity [2, 3], implying either that the HIV-1 RT RNase H domain contrasts in its ability to bind two divalent metal ions, or that the crystallographic data reflect specific use of Mn2+ and/or the doping technique employed. Metal binding stoichiometry has been determined for Mn2+ and the biologically more relevant Mg2+ cation by solution calorimetric studies of native and recombinant p66/p51 HIV-1 RT. Three Mn2+ ions bind to HIV-1 RT ape-enzyme: one at the DNA polymerase and two at the RNase H catalytic center, the latter being consistent with crystallographic results. However, only one Mg2+ ion is bound in the RNase H catalytic center. Several mechanistic implications arise from these results, including the possibility of mutually exclusive Mg2+ binding sites that might be occupied according to the specific reaction being catalyzed by the multifunctional RNase H domain. The occurrence of distinct binding stoichiometries for Mg2+ and Mn2+ to multifunctional enzymes has previously been reported [4]. C1 Ohio State Univ, Evans Lab Chem, Columbus, OH 43210 USA. Univ Hosp Cleveland, Ctr AIDS Res, Div Infect Dis, Cleveland, OH 44106 USA. Case Western Reserve Univ, Cleveland, OH 44106 USA. NCI, Resistance Mech Lab, HIV Drug Resistance Program, Frederick, MD 21702 USA. RP Cowan, JA (reprint author), Ohio State Univ, Evans Lab Chem, 100 W 18th Ave, Columbus, OH 43210 USA. FU NIAID NIH HHS [AI36219]; NIGMS NIH HHS [GM52263] NR 39 TC 35 Z9 38 U1 1 U2 7 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0949-8257 J9 J BIOL INORG CHEM JI J. Biol. Inorg. Chem. PD FEB PY 2000 VL 5 IS 1 BP 67 EP 74 DI 10.1007/s007750050009 PG 8 WC Biochemistry & Molecular Biology; Chemistry, Inorganic & Nuclear SC Biochemistry & Molecular Biology; Chemistry GA 295EZ UT WOS:000085954900008 PM 10766438 ER PT J AU Koenig, BW Mitchell, DC Konig, S Grzesiek, S Litman, BJ Bax, A AF Koenig, BW Mitchell, DC Konig, S Grzesiek, S Litman, BJ Bax, A TI Measurement of dipolar couplings in a transducin peptide fragment weakly bound to oriented photo-activated rhodopsin SO JOURNAL OF BIOMOLECULAR NMR LA English DT Article DE alignment; membrane protein; receptor; rhodopsin; transferred dipolar coupling; transducin ID LIQUID-CRYSTALLINE MEDIUM; G-PROTEIN RECEPTOR; PROJECTION STRUCTURE; ALPHA-SUBUNIT; NMR; INFORMATION; BINDING; FORM AB Rhodopsin-containing disks, isolated from rod outer segments of bovine retina, align at high magnetic fields with their membrane normal parallel to the magnetic field. After light-activation of rhodopsin, transient binding of the C-terminal transducin undecapeptide, selectively labeled with N-15 at Leu(5) and Gly(9), results in residual dipolar contributions to the (1)J(NH) splittings for these two residues. Both residues show (1)J(NH) splittings which are smaller than in the dark-adapted or rhodopsin-free sample, and return to their isotropic values at a rate determined by the decay of the meta II state of rhodopsin. The dipolar couplings indicate that in the bound state, N-H vectors of Leu(5) and Gly(9) make angles of 48 +/- 4 degrees and 40 +/- 8 degrees, respectively, with the disk normal. These `transferred' dipolar couplings potentially offer a useful method for studying the conformation and orientation of flexible, low affinity ligands when bound to oriented integral membrane receptors. C1 NIDDK, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. Tech Univ Darmstadt, Inst Biochem, D-64287 Darmstadt, Germany. Res Ctr Julich, Inst Biol Struct, D-52425 Julich, Germany. NIAAA, Lab Membrane Biophys & Biochem, NIH, Rockville, MD 20852 USA. NHLBI, Biophys Chem Lab, NIH, Bethesda, MD 20892 USA. RP Koenig, BW (reprint author), NIDDK, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. RI Koenig, Bernd/B-4315-2008; Konig, Simone/B-6504-2008 OI Koenig, Bernd/0000-0002-5300-6276; Konig, Simone/0000-0003-0672-7246 NR 32 TC 49 Z9 49 U1 0 U2 2 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 0925-2738 J9 J BIOMOL NMR JI J. Biomol. NMR PD FEB PY 2000 VL 16 IS 2 BP 121 EP 125 DI 10.1023/A:1008378523816 PG 5 WC Biochemistry & Molecular Biology; Spectroscopy SC Biochemistry & Molecular Biology; Spectroscopy GA 280YD UT WOS:000085130600003 PM 10723991 ER PT J AU Menaa, C Barsony, J Reddy, SV Cornish, J Cundy, T Roodman, GD AF Menaa, C Barsony, J Reddy, SV Cornish, J Cundy, T Roodman, GD TI 1,25-dihydroxyvitamin D-3 hypersensitivity of osteoclast precursors from patients with Paget's disease SO JOURNAL OF BONE AND MINERAL RESEARCH LA English DT Article DE Paget's disease; osteoclasts; precursors; 1,25-(OH)(2)D-3; vitamin D receptor ID VITAMIN-D-RECEPTOR; MARROW CULTURES; IMMUNOCYTOCHEMICAL LOCALIZATION; MULTINUCLEATED CELLS; GENE-EXPRESSION; PHORBOL ESTER; BONE-MARROW; HORMONE; BINDING; LIGAND AB Our previous studies suggested that increased osteoclast formation and activity in Paget's disease may be related in part to increased responsiveness of highly purified osteoclast precursors to 1,25-dihydroxyvitamin D-3 [1,25-(OH)(2)D-3]. However, the basis for this enhanced sensitivity to 1,25-(OH),D3 is unclear To address this question, we examined 24-hydroxylase and 1,25-(OH)(2)D-3 receptor (VDR) messenger RNA (mRNA) expression during human osteoclast differentiation from normal subjects and patients with Paget's disease in response to 1,25-(OH)(2)D-3 as well as VDR content and affinity, Reverse-transcription polymerase chain reaction (RT-PCR) analysis of granulocyte-macrophage colony-forming unit (GM-CFU), the earliest identifiable osteoclast precursor, derived from patients with Paget's disease demonstrated 24-hydroxylase mRNA expression in response to 1,25-(OH)(2)D-3 was induced at concentrations of 1,25-(OH)(2)D-3 that were at least one log less than that required for normal GM-CFU, VDR mRNA and VDR protein were detected in both immature and more differentiated osteoclast precursors, as well as in osteoclast-like multinucleated cells (MNCs), However, VDR expression was lower in MNCs than the mononuclear precursor cells, Osteoclast precursors and MNCs from patients with Paget's disease had levels of VDR expression similar to those of normal subjects but showed increased VDR affinity for 1,25-(OH)(2)D-3. Because the effects of 1,25-(OH)(2)D-3 are in part mediated by induction of expression of RANK ligand on marrow stromal cells, which in turn stimulates osteoclast formation, we examined expression of RANK ligand mRNA by marrow stromal cell lines derived from patients with Paget's disease and normal subjects in response to 1,25-(OH)(2)D-3. RT-PCR analysis showed no difference in sensitivity of marrow stromal cells to 1,25-(OH)(2)D-3 from normal subjects or patients with Paget's disease although the Paget's stromal cells expressed increased basal levels of RANK ligand mRNA, These results show that VDR protein is expressed in early and more differentiated osteoclast precursors, that expression levels of VDR decline with osteoclast differentiation, and that 1,25-(OH)(2)D-3 has direct effects on osteoclast precursors, The enhanced sensitivity to 1,25-(OH)(2)D-3 is an intrinsic property of osteoclast precursors from patients with Paget's disease that distinguishes them from normal osteoclast precursors, Furthermore, our results suggest that an increased affinity of VDR for 1,25-(OH)(2)D-3 may be responsible for the enhanced 1,25-(OH)(2)D-3 sensitivity of osteoclast precursors in patients with Paget's disease compared with normal subjects. C1 Univ Texas, Hlth Sci Ctr, Dept Med Hematol, San Antonio, TX 78284 USA. NIH, Bethesda, MD 20892 USA. Univ Auckland, Auckland 1, New Zealand. Audie Murphy Vet Adm Hosp, San Antonio, TX USA. RP Roodman, GD (reprint author), Res Hematol 151, 7400 Merton Minter Blvd, San Antonio, TX 78284 USA. FU NIA NIH HHS [AG13625]; NIAMS NIH HHS [AR41336, AR44603] NR 27 TC 52 Z9 54 U1 0 U2 3 PU AMER SOC BONE & MINERAL RES PI DURHAM PA PO BOX 2759, DURHAM, NC 27715-2759 USA SN 0884-0431 J9 J BONE MINER RES JI J. Bone Miner. Res. PD FEB PY 2000 VL 15 IS 2 BP 228 EP 236 DI 10.1359/jbmr.2000.15.2.228 PG 9 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 282HR UT WOS:000085212800006 PM 10703924 ER PT J AU Wortman, MJ Krachmarov, CP Kim, JH Gordon, RG Chepenik, LG Brady, JN Gallia, GL Khalili, K Johnson, EM AF Wortman, MJ Krachmarov, CP Kim, JH Gordon, RG Chepenik, LG Brady, JN Gallia, GL Khalili, K Johnson, EM TI Interaction of HIV-1 Tat with Pur alpha in nuclei of human glial cells: Characterization of RNA-mediated protein-protein binding SO JOURNAL OF CELLULAR BIOCHEMISTRY LA English DT Article DE AIDS; JC virus; JCV; progressive multifocal leukoencephalopathy; PML ID HUMAN-IMMUNODEFICIENCY-VIRUS; SINGLE-STRANDED-DNA; CARBOXY-TERMINAL DOMAIN; CENTRAL-NERVOUS-SYSTEM; IN-VITRO; STIMULATES TRANSCRIPTION; RETINOBLASTOMA PROTEIN; TRANSACTIVATOR TAT; CELLULAR PROTEIN; POLYMERASE-II AB A complex between the Tat protein, encoded by human immunodeficiency virus type 1 (HIV-1), and the cellular protein, Puree, has been implicated in activation of the late promoter of IC virus (JCV) and in enhancement of JCV DNA replication. JCV is the causative agent of progressive multifocal leukoencephalopathy (PML), an acquired immunodeficiency syndrome (AIDS) opportunistic infection of the brain. Pur alpha also binds the HIV-1 TAR RNA element and activates HIV-1 transcription, suggesting a role for RNA binding in the action of this protein. Using immunoelectron microscopy, we find that in human glial cells expressing both proteins, Tat and Pur alpha are colocalized in extranucleolar chromatin structural elements. The colocalized Pur alpha and Tat are nearly exclusively nuclear, although individual proteins can be seen in both nucleus and cytoplasm, suggesting a preferential tropism of the complex for the nucleus. Analysis of the interaction between purified proteins indicates that the Tat-Pur alpha interaction is strongly enhanced by the presence of RNA. Tat amino acids from 37-48 are essential for Tat binding. Residues 49-72, including the TAR RNA-binding domain, are critical for binding to Pura, while Cys(22), in the Tat transactivation domain, is responsible for an important global effect. Puree repeat II domains are involved in the interaction, and a polypeptide based on one such sequence inhibits binding. After RNase treatment of Pur alpha enhancement of Tat binding can be partially restored by addition of a single-stranded JCV DNA PUR element, to which Tat does not bind. The results indicate that the Tat-Pur alpha interaction is direct, rather than through an RNA link, and that RNA binding configures Pur alpha for optimal interaction with Tat. J. Cell. Biochem. 77:65-74, 2000. (C) 2000 Wiley-Liss, Inc. C1 CUNY Mt Sinai Sch Med, Dept Pathol, New York, NY 10029 USA. CUNY Mt Sinai Sch Med, Brookdale Ctr Mol & Dev Biol, New York, NY 10029 USA. NCI, Mol Virol Lab, NIH, Bethesda, MD 20892 USA. MCP Hahnemann Univ, Ctr Neurovirol, Philadelphia, PA 19102 USA. RP Johnson, EM (reprint author), CUNY Mt Sinai Sch Med, Dept Pathol, New York, NY 10029 USA. FU NIEHS NIH HHS [T32-ES07265]; NINDS NIH HHS [NS-35000] NR 45 TC 30 Z9 31 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0730-2312 J9 J CELL BIOCHEM JI J. Cell. Biochem. PD FEB PY 2000 VL 77 IS 1 BP 65 EP 74 DI 10.1002/(SICI)1097-4644(20000401)77:1<65::AID-JCB7>3.0.CO;2-U PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 286RE UT WOS:000085458300007 PM 10679817 ER PT J AU Dieudonne, SC Kerr, JM Xu, TS Sommer, B DeRubeis, AR Kuznetsov, SA Kim, IS Robey, PG Young, MF AF Dieudonne, SC Kerr, JM Xu, TS Sommer, B DeRubeis, AR Kuznetsov, SA Kim, IS Robey, PG Young, MF TI Differential display of human marrow stromal cells reveals unique mRNA expression patterns in response to dexamethasone SO JOURNAL OF CELLULAR BIOCHEMISTRY LA English DT Article DE differential display; human bone marrow stroma; dexamethasone; beta ig-h3 ID GROWTH-FACTOR-BETA; CALVARIAL OSTEOBLAST DIFFERENTIATION; MATRIX VESICLE PROTEINS; GENE-EXPRESSION; IN-VITRO; TGF-BETA; EXTRACELLULAR-MATRIX; MESSENGER-RNA; FIBRONECTIN; FIBROBLASTS AB Human bone marrow stromal cells (hBMSC) are pluripotent cells that have the ability to differentiate into bone, cartilage, hematopoietic-supportive stroma, and adipocytes in a process modulated by dexamethasone (DEX). To characterize changes in hBMSC in response to DEX, we carried out differential display experiments using hBMSC cultured for 1 week in the presence or absence of 10(-8) M DEX. When RNA from these cells was used for differential display, numerous cDNA bands were identified that were up-regulated and down-regulated by DEX. The cDNA bands were reamplified by PCR and directly used to screen an hBMSC cDNA library. Seven clones were isolated and characterized by DNA sequencing and found to encode the following genes: transforming growth factor-beta-induced gene product (beta ig-h3), calphobindin II, cytosolic thyroid-binding protein, 22-kDA smooth muscle protein (SM22), and the extracellular matrix proteins osteonectin/SPARC, type III collagen, and fibronectin. To confirm that these genes were regulated by EX, the cells were treated continuously with this hormone for periods ranging from 2 to 30 days, and steady-state mRNA levels were measured by Northern blot analysis. All genes showed some level of regulation by DEX. The most profound regulation by DEX was observed in the beta ig-h3 gene,which showed a relative 10-fold decrease in mRNA levels after 6 days of treatment. Interestingly, beta ig-h3 expression was not altered by DEX in fibroblasts from other human tissues, including thymus stromal fibroblasts, spleen stromal fibroblasts, and foreskin fibroblasts. In summary, differential display of DEX-treated hBMSC revealed unique patterns of gene expression and has provided new information about phenotypic changes that accompany the differentiation of hBMSC toward osteogenesis. Published 1999 Wiley-Liss, Inc.(dagger). C1 Kyung Pook Natl Univ, Sch Med, Dept Biochem, Taegu, South Korea. NIDR, Craniofacial & Skeletal Dis Branch, NIH, Bethesda, MD 20892 USA. RP Young, MF (reprint author), Natl Inst Craniofacial & Dental Res, Craniofacial & Skeletal Dis Branch, NIH, Bldg 30,Room 222, Bethesda, MD 20892 USA. RI Robey, Pamela/H-1429-2011; 김, 인산/I-8988-2014; Kim, In-San/D-3956-2017 OI Robey, Pamela/0000-0002-5316-5576; NR 67 TC 3 Z9 3 U1 0 U2 1 PU WILEY-LISS PI HOBOKEN PA DIV JOHN WILEY & SONS INC, 111 RIVER ST, HOBOKEN, NJ 07030 USA SN 0730-2312 J9 J CELL BIOCHEM JI J. Cell. Biochem. PD FEB PY 2000 VL 76 IS 2 BP 231 EP 243 DI 10.1002/(SICI)1097-4644(20000201)76:2<231::AID-JCB7>3.0.CO;2-X PG 13 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 273FQ UT WOS:000084697600007 ER PT J AU Tang, XM Beesley, JS Grinspan, JB Seth, P Kamholz, J Cambi, F AF Tang, XM Beesley, JS Grinspan, JB Seth, P Kamholz, J Cambi, F TI Cell cycle arrest induced by ectopic expression of p27 is not sufficient to promote oligodendrocyte differentiation SO JOURNAL OF CELLULAR BIOCHEMISTRY LA English DT Article DE oligodendrocytes; cell cycle; differentiation; p27; cdk2; cdk5 ID NEURONAL CDC2-LIKE KINASE; CENTRAL NERVOUS-SYSTEM; DEPENDENT KINASE-5; GROWTH-FACTOR; MUSCLE DIFFERENTIATION; MONOCLONAL-ANTIBODY; PROGENITOR CELLS; SKELETAL-MUSCLE; PDGF RECEPTORS; MICE LACKING AB Oligodendrocyte differentiation is-accompanied by dramatic changes in gene expression as well as cell cycle arrest. To determine whether cell cycle arrest is sufficient to induce the changes:in cell phenotype associated with differentiation, we inhibited oligodendrocyte precursor proliferation in vitro by overexpressing p27, a cyclin kinase inhibitor, using a recombinant adenovirus. Ectopic expression of p27 efficiently inhibited oligodendrocyte precursor cell division, even in the presence of exogenous mitogens, by blocking the activity of the cyclin-dependent kinase, cdk2. Although the cells had stopped dividing, they did not express galactocerebroside (GalC) or myelin: basic protein (MBP), changes associated with oligodendrocyte differentiation, suggesting that they had not differentiated. After removal of exogenous mitogens, however, adenovirus-expressing oligodendrocyte precursors differentiated with atemporal profile similar to that of control, uninfected oligodendrocytes, as indicated by expression of GalC and MBP. We conclude that cell cycle arrest is not sufficient to induce differentiation of dividing oligodendrocyte precursors, and that modulation of additional, as yet unknown, signaling pathways is required for this to occur. (C) 1999 Wiley-Liss, Inc. C1 Thomas Jefferson Univ, Dept Neurol, Philadelphia, PA 19107 USA. Childrens Hosp Philadelphia, Dept Res Neurol, Philadelphia, PA 19104 USA. NCI, Div Clin Sci, Med Branch, Bethesda, MD 20892 USA. Wayne State Univ, Sch Med, Dept Neurol, Detroit, MI 48201 USA. Wayne State Univ, Sch Med, Ctr Mol Med & Genet, Detroit, MI 48201 USA. RP Cambi, F (reprint author), Thomas Jefferson Univ, Dept Neurol, 1020 Locust St 391, Philadelphia, PA 19107 USA. NR 57 TC 1 Z9 1 U1 0 U2 0 PU WILEY-LISS PI HOBOKEN PA DIV JOHN WILEY & SONS INC, 111 RIVER ST, HOBOKEN, NJ 07030 USA SN 0730-2312 J9 J CELL BIOCHEM JI J. Cell. Biochem. PD FEB PY 2000 VL 76 IS 2 BP 270 EP 279 DI 10.1002/(SICI)1097-4644(20000201)76:2<270::AID-JCB10>3.0.CO;2-6 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 273FQ UT WOS:000084697600010 ER PT J AU Bruno, S Torrisi, R Costantini, M Baglietto, L Fontana, V Gatteschi, B Melioli, G Nicolo, G Curotto, A Malcangi, B Bruttini, GP Varaldo, M Bruzzi, P Decensi, A AF Bruno, S Torrisi, R Costantini, M Baglietto, L Fontana, V Gatteschi, B Melioli, G Nicolo, G Curotto, A Malcangi, B Bruttini, GP Varaldo, M Bruzzi, P Decensi, A TI Assessment of DNA flow cytometry as a surrogate end point biomarker in a bladder cancer chemoprevention trial SO JOURNAL OF CELLULAR BIOCHEMISTRY LA English DT Article DE bladder cancer; DNA flow cytometry; biomarker ID TRANSITIONAL-CELL CARCINOMA; PROGNOSTIC-SIGNIFICANCE; CYTOLOGY; HYBRIDIZATION; ANEUPLOIDY; DIAGNOSIS; WASHINGS AB Although conventional cytology represents the most widely performed cytometric analysis of bladder cancer cells, DNA flow cytometry has, over the past decade, been increasingly used to evaluate cell proliferation and DNA ploidy in cells from bladder washings. We have investigated whether DNA flow cytometry and conventional cytology of epithelial cells obtained from bladder washings provide reliable surrogate endpoint biomarkers in clinical chemoprevention trials. We used cytometric and clinical data from a chemoprevention trial of the synthetic retinoid Fenretinide on 99 patients with superficial bladder cancer. A total of 642 bladder washing specimens obtained from the patients at 4 month intervals was analyzed. Intra-individual agreement and correlation of flow cytometric DNA ploidy (diploid vs. aneuploid), DNA Index, Hyper-Diploid-Fraction (proportion of cells with DNA content higher than 2C), and conventional cytologic examination, as assessed by kappa statistics and Spearman's correlation test, were poor from baseline through 24 months. Moreover, no correlation was found between DNA ploidy and cytology at each rime point. The same results were obtained when the analyses were stratified by treatment group. In addition, the association between the results of bladder washing (by either DNA flow cytometry or cytology) and concomitant tumor recurrence was significant only for abnormal cytology, while neither biomarker was predictive of tumor recurrence at the subsequent visit. During the time of this study only four patients progressed to muscle-invasive bladder cancer, indicating the "low-risk" features of the patient population. We conclude that DNA flow cytometry and conventional cytology on epithelial cells obtained from bladder washings do not appear to provide suitable surrogate endpoint biomarkers during the early stages of bladder carcinogenesis. (C) 1999 Wiley-Liss, Inc. C1 Univ Genoa, Inst Human Anat, Dept Expt Med, Human Anat Sect, I-16132 Genoa, Italy. Natl Canc Inst, Cytometry Unit, I-16132 Genoa, Italy. Natl Canc Inst, Unit Med Oncol 2, I-16132 Genoa, Italy. European Inst Oncol, Chemoprevent Unit, I-20141 Milan, Italy. Natl Canc Inst, Unit Clin Epidemiol & Trials, I-16132 Genoa, Italy. European Inst Oncol, Div Epidemiol, I-20141 Milan, Italy. Natl Canc Inst, Biostat Unit, I-16132 Genoa, Italy. Natl Canc Inst, Unit Pathol, I-16132 Genoa, Italy. S Martino Hosp, Urol Clin, I-16132 Genoa, Italy. Gallino Hosp, Div Urol, I-16164 Genoa, Italy. Nervi Hosp, Div Urol, I-16167 Genoa, Italy. RP Bruno, S (reprint author), Univ Genoa, Inst Human Anat, Dept Expt Med, Human Anat Sect, Via Toni 14, I-16132 Genoa, Italy. RI costantini, massimo/G-1443-2012; Bruno, Silvia/B-8980-2013 NR 33 TC 0 Z9 0 U1 1 U2 2 PU WILEY-LISS PI HOBOKEN PA DIV JOHN WILEY & SONS INC, 111 RIVER ST, HOBOKEN, NJ 07030 USA SN 0730-2312 J9 J CELL BIOCHEM JI J. Cell. Biochem. PD FEB PY 2000 VL 76 IS 2 BP 311 EP 321 DI 10.1002/(SICI)1097-4644(20000201)76:2<311::AID-JCB14>3.0.CO;2-A PG 11 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 273FQ UT WOS:000084697600014 ER PT J AU Sheikh, MS Fornace, AJ AF Sheikh, MS Fornace, AJ TI Role of p53 family members in apoptosis SO JOURNAL OF CELLULAR PHYSIOLOGY LA English DT Review ID BREAST-CANCER CELLS; RADIATION-INDUCED APOPTOSIS; NECROSIS-FACTOR-ALPHA; TUMOR-SUPPRESSOR P53; BCL-X-L; GROWTH ARREST; GENE-EXPRESSION; TRANSCRIPTIONAL REPRESSION; P53-INDUCED APOPTOSIS; P53-RELATED PROTEIN AB p53-mediated apoptosis involves multiple mechanisms. A number of p53-regulated apoptosis-related genes have been identified. Some of these genes encode proteins that are important in controlling the integrity of mitochondria while the others code for membrane death receptors. p53 may also induce apoptosis by interfering with the growth factor-mediated survival signals. Although the transactivation-deficient p53 can induce apoptosis, evidence suggests that both the transcription-dependent and independent functions are: needed for full apoptotic activity. p73 and p63 are two other members of the p53 family that show homology to p53 in their respective transactivation, DNA-binding and oligomerization domains. Both p73 and p63 transactivate p53-regulated promoters and induce apoptosis. Evidence suggests that both p73 and p63 may mediate apoptosis via some of the same mechanisms that are utilized by p53. However, both p73 and p63 exhibit features that are different from those of p53. Hence, both p73 and p63 are predicted to mediate apoptosis via mechanisms that are completely distinct from those engaged by p53. Published 2000 Wiley-Liss, Inc.(dagger) C1 SUNY Hlth Sci Ctr, Dept Pharmacol, Syracuse, NY 13210 USA. NCI, Gene Response Sect, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Sheikh, MS (reprint author), SUNY Hlth Sci Ctr, Dept Pharmacol, 750 E Adams St, Syracuse, NY 13210 USA. RI Fornace, Albert/A-7407-2008 OI Fornace, Albert/0000-0001-9695-085X NR 124 TC 134 Z9 140 U1 0 U2 5 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0021-9541 J9 J CELL PHYSIOL JI J. Cell. Physiol. PD FEB PY 2000 VL 182 IS 2 BP 171 EP 181 DI 10.1002/(SICI)1097-4652(200002)182:2<171::AID-JCP5>3.0.CO;2-3 PG 11 WC Cell Biology; Physiology SC Cell Biology; Physiology GA 270LP UT WOS:000084537400005 PM 10623880 ER PT J AU Enkemann, SA Wang, RH Trumbore, MW Berger, SL AF Enkemann, SA Wang, RH Trumbore, MW Berger, SL TI Functional discontinuities in prothymosin alpha caused by caspase cleavage in apoptotic cells SO JOURNAL OF CELLULAR PHYSIOLOGY LA English DT Article ID RETINOBLASTOMA PROTEIN; POLY(ADP-RIBOSE) POLYMERASE; NUCLEAR-PROTEIN; HISTONE H1; IN-VITRO; LAMIN-A; DEATH; EXPRESSION; PROTEASES; GENE AB Our study examines the effect of apoptosis on prothymosin alpha, an abundant, nuclear protein intimately involved with proliferation of all mammalian cells. When HeLa cells were treated with actinomycin D, with etoposide, or with staurosporine following synchronization with hydroxyurea, they underwent apoptosis based on several specific criteria, including fragmentation of DNA and activation of specific caspases. Similarly treated NIH3T3 cells arrested and displayed no indicators of apoptosis. In HeLa, but not in NIH3T3 cells, prothymosin or levels declined precipitously and a truncated version of the protein was formed. The following observations implicate caspase activity: (1) The truncated polypeptide arose only in the treated HeLa cell cultures. (2) The appearance of the truncated polypeptide coincided with the activation of caspase 3 and the cleavage of poly(ADP-ribose) polymerase, a known caspase substrate. (3) Carbobenzoxy-DEVD-fluoromethylketone, a cell-permeable caspase 3 inhibitor, blocked cleavage and degradation of prothymosin alpha. (4) The same inhibitor, when added to mixed extracts of apoptotic and normal cells, prevented cleavage of intact prothymosin alpha. (5) Recombinant caspase 3 and, to a much lesser extent, caspase 7 truncated purified prothymosin alpha. (6) In HeLa cells, cleavage occurred at three overlapping caspase 3-like sites with the consensus sequence D-X-X-D and released 10 to 14 residues from the carboxyl terminus, including the core nuclear localization signal. Two immediate consequences of the cleavage were observed: truncated prothymosin alpha was no longer confined to the nucleus and it was deficient in phosphate. These data suggest that the disabling of prothymosin alpha is a significant event in apoptosis. Published 2000 Wiley-Liss, Inc.(dagger) C1 NCI, Sect Genes & Gene Prod, NIH, Bethesda, MD 20892 USA. RP Berger, SL (reprint author), NCI, Sect Genes & Gene Prod, NIH, Bldg 37,Room 5D-13, Bethesda, MD 20892 USA. NR 71 TC 23 Z9 26 U1 0 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0021-9541 J9 J CELL PHYSIOL JI J. Cell. Physiol. PD FEB PY 2000 VL 182 IS 2 BP 256 EP 268 DI 10.1002/(SICI)1097-4652(200002)182:2<256::AID-JCP15>3.0.CO;2-N PG 13 WC Cell Biology; Physiology SC Cell Biology; Physiology GA 270LP UT WOS:000084537400015 PM 10623890 ER PT J AU Shoaf, SE Carson, RE Hommer, D Williams, WA Higley, JD Schmall, B Herscovitch, P Eckelman, WC Linnoila, M AF Shoaf, SE Carson, RE Hommer, D Williams, WA Higley, JD Schmall, B Herscovitch, P Eckelman, WC Linnoila, M TI The suitability of [C-11]-alpha-methyl-L-tryptophan as a tracer for serotonin synthesis: Studies with dual administration of [C-11] and [C-14] labeled tracer SO JOURNAL OF CEREBRAL BLOOD FLOW AND METABOLISM LA English DT Article DE serotonin; positron emission tomography; rhesus monkeys; pharmacokinetics; brain; alpha-methyl-L-tryptophan ID POSITRON-EMISSION-TOMOGRAPHY; METHYL-L-TRYPTOPHAN; HUMAN BRAIN; SYNTHESIS INVIVO; SYNTHESIS RATES; RHESUS-MONKEYS; IN-VIVO; ALPHA-METHYL-L-TRYPTOPHAN; KINETICS; PET AB The tracer [C-11]-alpha-methyl-L-tryptophan (alpha MTP) has been used to measure brain serotonin synthesis rates with positron emission tomography (PET). To address questions about the accuracy of the kinetic model, [C-14]alpha MTP was used to directly measure conversion to [C-14]-alpha-methyl-serotonin (alpha M5HT) in monkeys that had been previously studied with PET and [C-11]alpha MTP. Four male, fasted, isonurane-anesthetized rhesus monkeys were studied with [C-11]aMTP and PET. Immediately after the initial 3-hour scan, a second dose of [C-11]alpha MTP was coinjected with 1 mCi of [C-14]alpha MTP, and additional PET data were collected. Approximately 90 minutes after the second aMTP administration. the animals were killed with an overdose of phenobarbital. and brain samples from 21 regions were taken and analyzed by HPLC. Minimal conversion of alpha MTP to alpha M5HT occurred; HPLC analysis of C-14 radioactivity showed that greater than 96% of the total counts were in fractions corresponding to the alpha MTP peak. Brain concentrations of serotonin, tryptophan, 5-hydroxyindole-3-acetic acid, and alpha MTP also were determined fluorometrically using exernal quantification. Patlak plots generated from PET images acquired over 3 hours showed no time period of linear increase, and final slopes were not significantly different from zero, consistent With the finding of minimal conversion to [C-14]alpha M5HT. These data indicate that in the 3-hour period after injection, [C-11]alpha MTP is acting predominantly as a tracer of tryptophan uptake, not serotonin synthesis. C1 NIAAA, DICBR, Clin Studies Lab, Bethesda, MD 20892 USA. NIH, PET Dept, Ctr Clin, Bethesda, MD 20892 USA. RP Shoaf, SE (reprint author), OAPI, 2440 Res Blvd, Rockville, MD 20850 USA. RI Carson, Richard/H-3250-2011 OI Carson, Richard/0000-0002-9338-7966 NR 23 TC 45 Z9 45 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0271-678X J9 J CEREBR BLOOD F MET JI J. Cereb. Blood Flow Metab. PD FEB PY 2000 VL 20 IS 2 BP 244 EP 252 PG 9 WC Endocrinology & Metabolism; Hematology; Neurosciences SC Endocrinology & Metabolism; Hematology; Neurosciences & Neurology GA 285JE UT WOS:000085385300004 PM 10698060 ER PT J AU Maital, SL Dromi, E Sagi, A Bornstein, MH AF Maital, SL Dromi, E Sagi, A Bornstein, MH TI The Hebrew Communicative Development Inventory: language specific properties and cross-linguistic generalizations SO JOURNAL OF CHILD LANGUAGE LA English DT Article; Proceedings Paper CT Biennial Meeting of the Society-for-Research-in-Child-Development CY APR 03-06, 1997 CL WASHINGTON, DC SP Soc Res Child Dev ID EARLY LEXICAL DEVELOPMENT; EARLY VOCABULARIES; CHECKLIST; MOTHER; ISRAEL; NOUNS; VERBS AB Cultural, linguistic, and developmental evidence was taken into consideration in constructing the HCDI, a Hebrew adaptation of the MCDI. The HCDI was then administered to a stratified sample of Israeli mothers of 253 toddlers aged I;6 to 2;0 (M = I;8.18). Hebrew results are presented and compared with scores from the original MCDI sample (Fenson, Dale, Reznick, Bares, Thal gt Pethick, 1994). The HCDI is a reliable and sensitive measure of lexical development and emergent grammar, capturing wide variability among Israeli toddlers. In comparison with English, the relation between vocabulary size and age, as well as the shape of the growth curves for nouns, predicate terms, and closed class words relative to size of lexicon, were strikingly similar. These results indicate that conclusions concerning cross-linguistic similarities can be best documented by using parallel methods of measurement. The HCDI results support the claim that early lexical development in Hebrew and in English follow remarkably similar development patterns, despite the typological differences Between the two target languages. C1 Univ Haifa, Fac Educ, IL-31999 Haifa, Israel. Tel Aviv Univ, IL-69978 Tel Aviv, Israel. Univ Texas, Dallas, TX 75230 USA. NICHHD, Bethesda, MD 20892 USA. RP Maital, SL (reprint author), Univ Haifa, Fac Educ, Mt Carmel, IL-31999 Haifa, Israel. EM maitals@construct.haifa.ac.il FU NICHD NIH HHS [R0I HD25975] NR 37 TC 55 Z9 55 U1 1 U2 10 PU CAMBRIDGE UNIV PRESS PI NEW YORK PA 32 AVENUE OF THE AMERICAS, NEW YORK, NY 10013-2473 USA SN 0305-0009 J9 J CHILD LANG JI J. Child Lang. PD FEB PY 2000 VL 27 IS 1 BP 43 EP 67 DI 10.1017/S0305000999004006 PG 25 WC Psychology, Developmental; Linguistics; Psychology, Experimental SC Psychology; Linguistics GA 295NB UT WOS:000085973200003 PM 10740967 ER PT J AU Nopoulos, P Berg, S Castellenos, FX Delgado, A Andreasen, NC Rapoport, JL AF Nopoulos, P Berg, S Castellenos, FX Delgado, A Andreasen, NC Rapoport, JL TI Developmental brain anomalies in children with attention-deficit hyperactivity disorder SO JOURNAL OF CHILD NEUROLOGY LA English DT Article; Proceedings Paper CT 35th Annual Meeting of the American-College-of-Neuropsychopharmacology CY DEC 09-13, 1996 CL SAN JUAN, PUERTO RICO SP Amer Coll Neuropsychopharmacol ID CAVUM SEPTI PELLUCIDI; GRAY-MATTER HETEROTOPIAS; CORPUS-CALLOSUM; POSTERIOR-FOSSA; SCHIZOPHRENIA; MRI; MALFORMATIONS; MORPHOLOGY; PREGNANCY AB The pathoetiology of attention-deficit hyperactivity disorder (ADHD) has been considered to be neurodevelopmental, yet the timing and processes involved are not clearly identified. Neurodevelopmental brain anomalies have been associated with a variety of psychiatric conditions. However, they have never been evaluated in a population of patients with ADHD. This study was designed to determine the frequency of specific developmental brain anomalies in a group, of children with ADHD (n = 85; mean age, 10.9 years) and healthy control children (n = 95; mean age, 11.7 years) by visually inspecting brain magnetic resonance imaging scans. Compared to controls, the ADHD group showed an increase in frequency of two developmental anomalies: (1) gray-matter heterotopia, a neuronal migration anomaly, in 2 of 85 patients versus 0 of 95 controls; and (2) posterior fossa abnormality (excess cerebrospinal fluid in the posterior fossa) in 8 of 85 patients versus 2 of 95 controls. There were no differences in frequency of enlarged cavum septi pellucidi between the two groups. These findings support and extend the idea that ADHD is of developmental origin, and further suggest that the timing of aberrant brain development could be in early gestation. C1 Univ Iowa Hosp & Clin, Mental Hlth Clin Res Ctr, Iowa City, IA 52242 USA. NIMH, Child Psychiat Branch, Bethesda, MD 20892 USA. RP Nopoulos, P (reprint author), Univ Iowa Hosp & Clin, Mental Hlth Clin Res Ctr, 2887 JPP,200 Hawkins Dr, Iowa City, IA 52242 USA. FU NIMH NIH HHS [MH31593, MH40856, MHCRC 43271] NR 49 TC 21 Z9 21 U1 2 U2 2 PU DECKER PERIODICALS INC PI HAMILTON PA 4 HUGHSON STREET SOUTH PO BOX 620, LCD 1, HAMILTON, ONTARIO L8N 3K7, CANADA SN 0883-0738 J9 J CHILD NEUROL JI J. Child Neurol. PD FEB PY 2000 VL 15 IS 2 BP 102 EP 108 DI 10.1177/088307380001500208 PG 7 WC Clinical Neurology; Pediatrics SC Neurosciences & Neurology; Pediatrics GA 285EE UT WOS:000085376100008 PM 10695895 ER PT J AU Mericq, MV Eggers, M Avila, A Cutler, GB Cassorla, F AF Mericq, MV Eggers, M Avila, A Cutler, GB Cassorla, F TI Near final height in pubertal growth hormone (GH)-deficient patients treated with GH alone or in combination with luteinizing hormone-releasing hormone analog: Results of a prospective, randomized trial SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID LONG-TERM TREATMENT; ADULT HEIGHT; PRECOCIOUS PUBERTY; DEFICIENCY; CHILDREN; DESLORELIN; AGONIST; DELAY AB To study the effects of delaying puberty in GH-deficient (GHD) children, we studied 21 GHD (9 boys, 14 girls), treatment-naive, pubertal patients in a prospective, randomized trial. Their chronological age was 14.3 +/- 1.6 yr, and their bone age was 11.3 +/- 1.1 yr (mean +/- SD) at the beginning of the study. Four patients who developed hypogonadotropic hypogonadism were subsequently excluded from the study. Patients were randomly assigned to receive GH; LH-releasing hormone analog(LHRH-A) (n = 7), or GH alone (n = 10). GH and LHRH-A treatment started simultaneously in each patient. GH (Nutropin) was administered at a dose of 0.1 U/kg . day sc, until patients reached a bone age (BA) of 14 yr in girls and 16 yr in boys, and LHRH-A (Lupron depot) was administered at a dose of 300 mu g/kg . every 28 days in during 3 yr. We defined GH deficiency as patients with a growth velocity less than 4 cm/yr, BA delay more than 1 yr in relationship to chronological age, GH response to two stimulation tests less than 7 mu g/L, associated with low serum insulin-like growth factor I and insulin-like growth factor binding protein 3 levels. Statistical analysis was performed by ANOVA or Kruskall Wallis when variances were not homogeneous. We observed a significant decrease in the rate of BA maturation in the group treated with GH + LHRH-A (1.5 +/- 0.2 yr) compared with the group treated with GH alone (4.2 +/- 0.5 yr) during the 3 years of LHRH-A therapy (P < 0.05). This delay in BA maturation produced a significant gain in final height in the group treated with GH+LHRH-A, which reached - 1.3 +/- 0.5 so score compared with -2.7 +/- 0.3 SD score (P < 0.05) in the group treated with GH alone. These results indicate that delaying puberty with LHRH-A in GHD children during treatment with GH increases final height. C1 Univ Chile, Inst Invest Materno Infantil, Santiago, Chile. NICHD, DEB, NIH, Bethesda, MD 20892 USA. RP Mericq, MV (reprint author), Univ Chile, Inst Invest Materno Infantil, Casilla 226-3, Santiago, Chile. NR 22 TC 54 Z9 57 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 FEB PY 2000 VL 85 IS 2 BP 569 EP 573 DI 10.1210/jc.85.2.569 PG 5 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 285EX UT WOS:000085377700020 PM 10690857 ER PT J AU Schuppert, F Ehrenthal, D Frilling, A Suzuki, K Napolitano, G Kohn, LD AF Schuppert, F Ehrenthal, D Frilling, A Suzuki, K Napolitano, G Kohn, LD TI Increased major histocompatibility complex (MHC) expression in nontoxic goiters is associated with iodide depletion, enhanced ability of the follicular thyroglobulin to increase MHC gene expression, and thyroid autoantibodies SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID THYROTROPIN RECEPTOR GENE; TRANSCRIPTION FACTOR-I; GRAVES-DISEASE; ASIALOGLYCOPROTEIN RECEPTOR; MULTINODULAR GOITER; CELLS; RECOGNITION; MEMBRANES; GROWTH; IMMUNOGLOBULINS AB Recent studies suggest that thyroglobulin (TG) accumulated in the follicular lumen of colloid nodular goiters can increase major histocompatibility complex (MHC) class I gene expression in FRTL-5 thyrocytes. Iodide deficiency, also present in these patients, was separately suggested to enhance thyroidal MHC class I and class LI gene expression in vivo and in vitro. To test the clinical relevance of these observations, we examined 41 nontoxic goiters surgically removed from patients who had compression problems. Northern analysis revealed that there was a mean 3.9-fold increase in MHC class I expression and a 8.3-fold increase in class II expression by comparison to 9 normal glands. In situ hybridization showed that thyrocytes were the main source of class I and class II transcripts; histological examination revealed that lymphocytic infiltration was minimal to nonexistent. The iodine content of the 41 nontoxic goiters was significantly lower than in normal glands, consistent with increased MHC class I and class II. There is also a profound accumulation of TG in the follicles of the nontoxic goiters, and TG purified from the follicles of these glands increased MHC class I gene expression in FRTL-5 thyroid cells significantly more than TG from normal glands per mg protein. Nearly all patients with nontoxic goiter had low, but significantly elevated, levels of antibodies against thyroid peroxidase and/or against TG in their sera compared with those in normal individuals. Moreover, there was a positive correlation between the titer of the serum antibodies against thyroid peroxidase and against TG and MHC class I and class II expression in the thyroid. The data support the possibility that the TG accumulated in the follicular lumen of nontoxic goiters together with relative iodine deficiency contributes to increased MHC expression in thyroid cells in vivo and that increased MHC gene expression contributes to the ability of thyroid antigens to trigger an autoimmune reaction. C1 Hannover Med Sch, Dept Clin Endocrinol, D-30625 Hannover, Germany. Univ Essen Gesamthsch, Dept Surg, D-45122 Essen, Germany. Univ G DAnnunzio, Chair Endocrinol, I-66100 Chieti, Italy. NIDDKD, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. RP Schuppert, F (reprint author), Hannover Med Sch, Dept Clin Endocrinol, Carl Neuberg Str 1, D-30625 Hannover, Germany. NR 40 TC 6 Z9 8 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 FEB PY 2000 VL 85 IS 2 BP 858 EP 867 DI 10.1210/jc.85.2.858 PG 10 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 285EX UT WOS:000085377700066 PM 10690902 ER PT J AU Murray, SA Davis, K Fishman, LM Bornstein, SR AF Murray, SA Davis, K Fishman, LM Bornstein, SR TI alpha(1) connexin 43 gap junctions are decreased in human adrenocortical tumors SO JOURNAL OF CLINICAL ENDOCRINOLOGY & METABOLISM LA English DT Article ID C6 GLIOMA-CELLS; ADRENAL-GLAND; IN-VIVO; EXPRESSION; COMMUNICATION; PROTEIN; GROWTH; GENES; RAT; MODULATION AB Gap junctional communication disorders have been implicated in the etiology of benign and malignant tumors. Understanding the type, distribution, and frequency of gap junctions in adrenal disorders should provide insight into the role of gap junctions in adrenal carcinogenesis as well as information that may be useful in developing improved diagnosis and treatment of adrenal diseases. Using immunocytochemical techniques, we have characterized and compared a, connexins 43 gap junction protein levels in normal adrenal glands to those in benign and malignant adrenocortical human tumors. In addition, gap junction protein levels were studied in a human adrenal cancer cell line (H295). In both normal and neoplastic adrenal tissues, only alpha(1) connexin 43 could be detected, whereas beta(1) connexin 32 and beta(2) connexin 26 were not found. In the normal adrenal gland, the tons fasciculata was demonstrated to have the highest number of gap junctions per cell (mean +/- SEM, 13.78 +/- 1.93). In contrast, in benign adrenocortical adenomas, the number of gap junctions per cell compared to that detected in normal adrenal glands was significantly reduced (mean +/- SEM, 4.6 +/- 1.17; P less than or equal to 0.05), and the lowest number was found in malignant adrenocortical tumors(1.42 +/- 0.58; P less than or equal to 0.05). Similarly, there were few or no a, connexin 43 gap junctions in the H295 population. There was a progressive decrease in gap junction plaques in adrenocortical cancer cell populations compared to those in normal cell populations. Therefore, analysis of gap junction protein may be helpful for the differential diagnosis of benign and malignant adrenal tumors. The induction of gap junctions in malignant cells may provide a novel therapeutic strategy for adrenal cancer. C1 Univ Pittsburgh, Sch Med, Dept Cell Biol & Physiol, Pittsburgh, PA 15261 USA. Univ Miami, Sch Med, Dept Med, Div Endocrinol, Miami, FL 33125 USA. Vet Affairs Med Ctr, Med Res Serv, Miami, FL 33125 USA. NICHHD, Dept Endocrinol Branch, NIH, Bethesda, MD 20892 USA. RP Murray, SA (reprint author), Univ Pittsburgh, Sch Med, Dept Cell Biol & Physiol, Pittsburgh, PA 15261 USA. NR 43 TC 27 Z9 35 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 FEB PY 2000 VL 85 IS 2 BP 890 EP 895 DI 10.1210/jc.85.2.890 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 285EX UT WOS:000085377700071 PM 10690907 ER PT J AU Gavrilova, O Marcus-Samuels, B Graham, D Kim, JK Shulman, GI Castle, AL Vinson, C Eckhaus, M Reitman, ML AF Gavrilova, O Marcus-Samuels, B Graham, D Kim, JK Shulman, GI Castle, AL Vinson, C Eckhaus, M Reitman, ML TI Surgical implantation of adipose tissue reverses diabetes in lipoatrophic mice SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article ID PARTIAL LIPODYSTROPHY; INSULIN-RESISTANCE; GLUCOSE-TOLERANCE; OBESITY; GENE; MUSCLE; MOUSE; LEPTIN; RATS AB In lipoatrophic diabetes, a lack of fat is associated with insulin resistance and hyperglycemia. This is in striking contrast to the usual association of diabetes with obesity. To understand the underlying mechanisms, we transplanted adipose tissue into A-ZIP/F-1 mice, which have a severe form of lipoatrophic diabetes. Transplantation of wild-type fat reversed the hyperglycemia, dramatically lowered insulin levels, and improved muscle insulin sensitivity, demonstrating that the diabetes in A-ZIP/F-1 mice is caused by the lack of adipose tissue. All aspects of the A-ZIP/F-1 phenotype including hyperphagia, hepatic steatosis, and somatomegaly were either partially or completely reversed. However, the improvement in triglyceride and FFA levels was modest. Donor fat taken from parametrial and subcutaneous sites was equally effective in reversing the phenotype. The beneficial effects of transplantation were dose dependent and required near-physiological amounts of transplanted fat. Transplantation of,genetically modified fat into A-ZIP/F-1 mice is a new and powerful technique for studying adipose physiology and the metabolic and endocrine communication between adipose tissue and the rest of the body. C1 NIDDKD, NIH, Diabet Branch, Bethesda, MD 20892 USA. Yale Univ, Sch Med, Howard Hughes Med Inst, New Haven, CT 06536 USA. Yale Univ, Sch Med, Dept Internal Med, New Haven, CT 06536 USA. NCI, NIH, Biochem Lab, Bethesda, MD 20892 USA. NCI, NIH, Vet Resources Program, Off Res Serv, Bethesda, MD 20892 USA. RP Reitman, ML (reprint author), NIDDKD, NIH, Diabet Branch, Bldg 10,Room 8N-250, Bethesda, MD 20892 USA. RI Reitman, Marc/B-4448-2013 OI Reitman, Marc/0000-0002-0426-9475 FU NIDDK NIH HHS [R01 DK040936, R01 DK080756] NR 31 TC 391 Z9 400 U1 0 U2 5 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA ROOM 4570 KRESGE I, 200 ZINA PITCHER PLACE, ANN ARBOR, MI 48109-0560 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD FEB PY 2000 VL 105 IS 3 BP 271 EP 278 DI 10.1172/JCI7901 PG 8 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 282FP UT WOS:000085207600005 PM 10675352 ER PT J AU Morteau, O Morham, SG Sellon, R Dieleman, LA Langenbach, R Smithies, O Sartor, RB AF Morteau, O Morham, SG Sellon, R Dieleman, LA Langenbach, R Smithies, O Sartor, RB TI Impaired mucosal defense to acute colonic injury in mice lacking cyclooxygenase-1 or cyclooxygenase-2 SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article ID NONSTEROIDAL ANTIINFLAMMATORY DRUGS; DEXTRAN SULFATE SODIUM; EXPERIMENTAL COLITIS; PERITONEAL-MACROPHAGES; ULCERATIVE-COLITIS; GENE DISRUPTION; PROSTAGLANDINS; INFLAMMATION; INHIBITION; STOMACH AB To investigate roles in intestinal inflammation for the 2 cyclooxygenase (COX) isoforms, we determined susceptibility to spontaneous and induced acute colitis in mice lacking either the COX-1 or COX-2 isoform. We treated wild-type, COX-1(-/-), COX-2(-/-), and heterozygous mice with dextran sodium sulfate (DSS) to provoke acute colonic inflammation, and we quantified tissue damage, prostaglandin (PG) E-2, and interleukin-1 beta. No spontaneous gastrointestinal inflammation was detected in mice homozygous for either mutation, despite almost undetectable basal intestinal PGE(2) production in COX-1-/- mice. Both COX-1(-/-) and COX-2(-/-) mice showed increased susceptibility to a low-dose of DSS that caused mild colonic epithelial injury in wild-type mice. COX-2(-/-) mice were more susceptible than COX-1(-/-) mice, and selective pharmacologic blockade of COX-2 potentiated injury in COX-1(-/-) mice. At a high dose, DSS treatment was fatal to 50% of the animals in each mutant group, but all wild-type mice survived. DSS treatment increased PGE(2) intestinal secretion in all groups except COX-2(-/-) mice. These results demonstrate that COX-1 and COX-2 share a crucial role in the defense of the intestinal mucosa (with inducible COX-2 being perhaps more active during inflammation) and that neither isoform is essential in maintaining mucosal homeostasis in the absence of injurious stimuli. C1 Univ N Carolina, Div Digest Dis, Dept Med, Chapel Hill, NC 27599 USA. Univ N Carolina, Dept Pathol, Chapel Hill, NC 27599 USA. N Carolina State Univ, Sch Vet Med, Dept Med, Raleigh, NC 27606 USA. NIEHS, Res Triangle Pk, NC 27709 USA. RP Sartor, RB (reprint author), Univ N Carolina, Div Digest Dis, Dept Med, Campus Box 7080,Room 030,Glaxo Bldg, Chapel Hill, NC 27599 USA. FU NIDDK NIH HHS [DK 53347, DK 34987, DK 40249, P30 DK034987, R01 DK040249, R01 DK053347] NR 49 TC 203 Z9 207 U1 0 U2 2 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA ROOM 4570 KRESGE I, 200 ZINA PITCHER PLACE, ANN ARBOR, MI 48109-0560 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD FEB PY 2000 VL 105 IS 4 BP 469 EP 478 DI 10.1172/JCI6899 PG 10 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 286LD UT WOS:000085446700010 PM 10683376 ER PT J AU Blagosklonny, MV Robey, R Bates, S Fojo, T AF Blagosklonny, MV Robey, R Bates, S Fojo, T TI Pretreatment with DNA-damaging agents permits selective killing of checkpoint-deficient cells by microtubule-active drugs SO JOURNAL OF CLINICAL INVESTIGATION LA English DT Article ID HUMAN CANCER-CELLS; ANTICANCER AGENTS; INDUCED APOPTOSIS; P53; ARREST; TAXOL; P21; THERAPY; GROWTH; INHIBITOR AB Cell-cycle checkpoint mechanisms, including the p53- and p21-dependent G(2) arrest that follows DNA damage, are often lost during tumorigenesis. We have exploited the ability of DNA-damaging drugs to elicit this checkpoint, and we show here that such treatment allows microtubule drugs, which cause cell death secondary to mitotic arrest, to kill checkpoint-deficient tumor cells while sparing checkpoint-competent cells. Low doses of the DNA-damaging drug doxorubicin cause predominantly Gz arrest without killing HCT116 cells that harbor wt p53. Doxorubicin treatment prevented mitotic arrest, Bcl-2 phosphorylation, and cell death caused by paclitaxel, epothilones, and vinblastine. In contrast, doxorubicin enhanced cytotoxicity of FR901228, an agent that does not affect microtubules. Low doses of doxorubicin did not arrest p21-deficient clones of HCT116 cells and did not protect these cells from cytotoxicity caused by microtubule drugs, but cells in which p21 expression was restored enjoyed partial protection under these conditions. Moreover, in p53-deficient clones of HCT116 cells doxorubicin did not induce either p53 or p21 and provided no protection against paclitaxel-induced cytotoxicity. Therefore, (a) p53-dependent p21 induction caused by doxorubicin protects from microtubule drug-induced cytotoxicity, and (b) pretreatment with cytostatic doses of DNA-damaging drugs before treatment with microtubule drugs results in selective cytotoxicity to cancer cells with defective p53/p21-dependent checkpoint. C1 NCI, Med Branch, NIH, Bethesda, MD 20892 USA. RP Blagosklonny, MV (reprint author), NCI, Med Branch, NIH, Bldg 10,R12N226, Bethesda, MD 20892 USA. NR 38 TC 95 Z9 96 U1 0 U2 1 PU AMER SOC CLINICAL INVESTIGATION INC PI ANN ARBOR PA ROOM 4570 KRESGE I, 200 ZINA PITCHER PLACE, ANN ARBOR, MI 48109-0560 USA SN 0021-9738 J9 J CLIN INVEST JI J. Clin. Invest. PD FEB PY 2000 VL 105 IS 4 BP 533 EP 539 DI 10.1172/JCI8625 PG 7 WC Medicine, Research & Experimental SC Research & Experimental Medicine GA 286LD UT WOS:000085446700017 PM 10683383 ER PT J AU Woods, GL Bergmann, JS Witebsky, FG Fahle, GA Boulet, B Plaunt, M Brown, BA Wallace, RJ Wanger, A AF Woods, GL Bergmann, JS Witebsky, FG Fahle, GA Boulet, B Plaunt, M Brown, BA Wallace, RJ Wanger, A TI Multisite reproducibility of Etest for susceptibility testing of Mycobacterium abscessus, Mycobacterium chelonae, and Mycobacterium fortuitum SO JOURNAL OF CLINICAL MICROBIOLOGY LA English DT Article ID RAPIDLY GROWING MYCOBACTERIA; ANTIMICROBIAL SUSCEPTIBILITY; CLARITHROMYCIN; RESISTANCE; INFECTIONS; SUBGROUPS; ORGANISM; AMIKACIN; FEATURES AB A multicenter study was conducted to assess the inter- and intralaboratory reproducibility of the Etest for susceptibility. testing of the rapidly growing mycobacteria. The accuracy also was evaluated by comparing Etest results to those obtained by broth microdilution, Ten isolates (four of the Mycobacterium fortuitum group, three of Mycobacterium abscessus, and three of Mycobacterium chelonae) were tested against amikacin, cefoxitin, ciprofloxacin, clarithromycin, doxycycline, imipenem, and trimethoprim-sulfamethoxazole in each of four laboratories, At each site, isolates were tested three times on each of three separate days (nine testing events per isolate) using common lots of media and Etest strips. Interlaboratory agreement among MICs (i.e., mode +/- I twofold dilution) varied for the different drug-isolate combinations and overall was best for trimethoprimsulfamethoxazole (75% for one isolate and 100% for all others), followed by doxycycline and ciprofloxacin. Interlaboratory agreement based on interpretive category also varied and overall was best for doxycycline (100% for all isolates, followed by trimethoprim-sulfamethoxazole and ciprofloxacin. Interlaboratory reproducibility among MICs was most variable for imipenem, and agreement by interpretive category was lowest for imipenem and amikacin, Modal Etest MICs agreed with those by broth microdilution only for doxycycline and the sulfonamides, For all other drugs, the modal MICs by the two methods differed by more than +/- I twofold dilution for one or more isolates. In all cases, the Etest MIC was higher and would hare caused reports of false resistance. In summary, the Etest in this evaluation did not perform as well as broth microdilution for susceptibility testing of the rapidly growing mycobacteria, It was problematic for most species and drugs, primarily because of a trailing endpoint and/or high MICs compared to broth. Its use will necessitate further investigation, including determination of the optimal medium and incubation conditions and clarification of endpoint interpretation. C1 Univ Texas, Med Branch, Dept Pathol, Galveston, TX 77555 USA. NIH, WG Magnuson Clin Ctr, Dept Clin Pathol, Microbiol Serv, Bethesda, MD 20892 USA. Univ Texas, Sch Med, Dept Pathol, Houston, TX 77030 USA. StatProbe, Ann Arbor, MI 48108 USA. Univ Texas, Ctr Hlth, Dept Microbiol, Tyler, TX 75710 USA. RP Woods, GL (reprint author), Univ Texas, Med Branch, Dept Pathol, Galveston, TX 77555 USA. NR 21 TC 48 Z9 49 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 USA SN 0095-1137 J9 J CLIN MICROBIOL JI J. Clin. Microbiol. PD FEB PY 2000 VL 38 IS 2 BP 656 EP 661 PG 6 WC Microbiology SC Microbiology GA 281XR UT WOS:000085187200033 PM 10655363 ER PT J AU Soini, H Pan, X Amin, A Graviss, EA Siddiqui, A Musser, JM AF Soini, H Pan, X Amin, A Graviss, EA Siddiqui, A Musser, JM TI Characterization of Mycobacterium tuberculosis isolates from patients in Houston, Texas, by spoligotyping SO JOURNAL OF CLINICAL MICROBIOLOGY LA English DT Article ID BOVIS STRAINS; DIFFERENTIATION; POLYMORPHISM; IDENTIFICATION; EPIDEMIOLOGY; TRANSMISSION; DIAGNOSIS; GENOTYPE; MICROTI AB Mycobacterium tuberculosis isolates (n = 1,429) from 1,283 patients collected as part of an ongoing population-based tuberculosis epidemiology study in Houston, Texas, were analyzed by spoligotyping and IS6110 profiling. The isolates were also assigned to one of three major genetic groups on the basis of nucleotide polymorphisms located at codons 463 and 95 in the genes (katG and gyrA) encoding catalase-peroxidase and the A subunit of DNA gyrase, respectively. A total of 225 spoligotypes were identified in the 1,429 isolates. There were 54 spoligotypes identified among 713 isolates (n = 623 patients) assigned to 73 IS6110 clusters. In addition, among 716 isolates (n = 660 patients) with unique IS6110 profiles, 200 spoligotypes were identified. No changes were observed either in the IS6110 profile or in the spoligotype for the 281 isolates collected sequentially from 133 patients. Five instances in which isolates with slightly different spoligotypes had the same IS6110 profile were identified, suggesting that in rare cases isolates with different spoligotypes can be clonally related. Spoligotypes correlated extremely well with major genetic group designations. Only three very similar spoligotypes were shared by isolates from genetic groups 2 and 3, and none was shared by group 1 and group 2 organisms or by group 1 end group 3 organisms. All organisms belonging to genetic groups 2 and 3 failed to hybridize with spacer probes 33 to 36, Taken together, the results support the existence of three distinct genetic groups of ill. tuberculosis organisms and provide new information about the relationship between IS6110 profiles, spoligotypes, and major genetic groups of hi. tuberculosis. C1 Baylor Coll Med, Inst Study Human Bacterial Pathogenesis, Dept Pathol, Houston, TX 77030 USA. RP Musser, JM (reprint author), NIAID, Rocky Mt Lab, Lab Human Bacterial Pathogenesis, NIH, 903 S 4th St, Hamilton, MT 59840 USA. FU NIDA NIH HHS [DA09238] NR 29 TC 101 Z9 101 U1 0 U2 2 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 USA SN 0095-1137 J9 J CLIN MICROBIOL JI J. Clin. Microbiol. PD FEB PY 2000 VL 38 IS 2 BP 669 EP 676 PG 8 WC Microbiology SC Microbiology GA 281XR UT WOS:000085187200035 PM 10655365 ER PT J AU Bemis, DA Krahwinkel, DJ Bowman, LA Mondon, P Kwon-Chung, KJ AF Bemis, DA Krahwinkel, DJ Bowman, LA Mondon, P Kwon-Chung, KJ TI Temperature-sensitive strain of Cryptococcus neoformans producing hyphal elements in a feline nasal granuloma SO JOURNAL OF CLINICAL MICROBIOLOGY LA English DT Article ID CATS AB We report the isolation of a temperature-sensitive, serotype A, mating type alpha strain of Cryptococcus neoformans from a case of nasal cryptococcosis in a cat. The strain grew extremely slowly at 35 degrees C and failed to grow at 37 degrees C in vitro. Histopathological sections of the infected tissue revealed Feast cells producing hyphae up to several hundred micrometers in length, in addition to numerous encapsulated yeast cells typical of C. neoformans. The cultures grown on yeast extract-peptone glucose agar at 35 degrees C also produced some yeast cells with germ tube-like hyphal elements up to 100 mu m in length. C1 Univ Tennessee, Coll Vet Med, Dept Comparat Med, Knoxville, TN 37996 USA. Univ Tennessee, Coll Vet Med, Dept Small Anim Clin Sci, Knoxville, TN 37996 USA. Univ Tennessee, Coll Vet Med, Dept Pathol, Knoxville, TN 37996 USA. NIAID, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. RP Bemis, DA (reprint author), Univ Tennessee, Coll Vet Med, Dept Comparat Med, 2407 River Dr, Knoxville, TN 37996 USA. NR 9 TC 14 Z9 15 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1325 MASSACHUSETTS AVENUE, NW, WASHINGTON, DC 20005-4171 USA SN 0095-1137 J9 J CLIN MICROBIOL JI J. Clin. Microbiol. PD FEB PY 2000 VL 38 IS 2 BP 926 EP 928 PG 3 WC Microbiology SC Microbiology GA 281XR UT WOS:000085187200089 PM 10655419 ER PT J AU Takimoto, CH Morrison, G Harold, N Quinn, M Monahan, BP Band, RA Cottrell, J Guemei, A Llorens, V Hehman, H Ismail, AS Flemming, D Gosky, DM Hirota, H Berger, SJ Berger, NA Chen, AP Shapiro, JD Arbuck, SG Wright, J Hamilton, JM Allegra, CJ Grem, JL AF Takimoto, CH Morrison, G Harold, N Quinn, M Monahan, BP Band, RA Cottrell, J Guemei, A Llorens, V Hehman, H Ismail, AS Flemming, D Gosky, DM Hirota, H Berger, SJ Berger, NA Chen, AP Shapiro, JD Arbuck, SG Wright, J Hamilton, JM Allegra, CJ Grem, JL TI Phase I and pharmacologic study of irinotecan administered as a 96-hour infusion weekly to adult cancer patients SO JOURNAL OF CLINICAL ONCOLOGY LA English DT Article ID ACTIVE METABOLITE SN-38; CAMPTOTHECIN DERIVATIVE IRINOTECAN; EVERY 3 WEEKS; COLORECTAL-CANCER; CPT-11; TRIAL; PHARMACOKINETICS; PHARMACODYNAMICS; INHIBITORS; TOPOTECAN AB Purpose: We conducted a phase I and pharmacologic study of a weekly 96-hour infusion of irinotecan ta determine the maximum-tolerated dose, define the toxicity profile, and characterize the clinical pharmacology of irinotecan and its metabolites. Patients and Methods: In 26 adult patients with solid tumors, the duration and dose rate of infusion were escalated in new patients until toxicity was observed. Results: In 11 patients who were treated with irinotecan at 12.5 mg/m(2)/d for 4 days weekly for 2 of 3 weeks, dose-limiting grade 3 diarrhea occurred in three patients and grade 3 thrombocytopenia occurred in two patients. The recommended phase II dose is 10 mg/m2/d for 4 days given weekly for 2 of 3 weeks. At this dose, the steady-state plasma concentration (Css) of total SN-38 (the active metabolite of irinotecan) was 6.42 +/- 1.10 nmol/L, and the Css of total irinotecan was 28.60 +/- 17.78 nmol/L. No patient experienced grade 3 or 4 neutropenia during any cycle. All other toxicities were mild to moderate. The systemic exposure to SN-38 relative to irinotecan was greater than anticipated, with a molar ratio of the area under the concentration curve (AUC) of SN-38 to irinotecan of 0.24 +/- 0.08. One objective response lasting 12 months in duration was observed in a patient with metastatic colon cancer. Conclusion: The recommended phase II dose of irinotecan of 10 mg/m(2)/d for 4 days weekly for 2 of 3 weeks was extremely well tolerated. Further efficacy testing of this pharmacologic strategy of administering intermittent low doses of irinotecan is warranted. J Clin Oncol 18:659-667. (C) 2000 by American Society of Clinical Oncology. C1 NCI, Med Branch, Natl Naval Med Ctr, Dept Dev Therapeut,Div Clin Sci, Bethesda, MD 20889 USA. Case Western Reserve Univ, Sch Med, Canc Res Ctr, Cleveland, OH USA. NCI, Investigat Drug Branch, Canc Therapy Evaluat Program, Div Canc Treatment & Diag, Bethesda, MD 20889 USA. Natl Naval Med Ctr, Dept Radiol, Bethesda, MD USA. RP Takimoto, CH (reprint author), NCI, Med Branch, Natl Naval Med Ctr, Dept Dev Therapeut,Div Clin Sci, Bldg 8,Rm 5101,8901 Wisconsin Ave, Bethesda, MD 20889 USA. NR 32 TC 48 Z9 48 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0732-183X J9 J CLIN ONCOL JI J. Clin. Oncol. PD FEB PY 2000 VL 18 IS 3 BP 659 EP 667 PG 9 WC Oncology SC Oncology GA 281ZX UT WOS:000085192300024 PM 10653882 ER PT J AU Eisenhauer, EA O'Dwyer, PJ Christian, M Humphrey, JS AF Eisenhauer, EA O'Dwyer, PJ Christian, M Humphrey, JS TI Phase I clinical trial design in cancer drug development SO JOURNAL OF CLINICAL ONCOLOGY LA English DT Article ID CONTINUAL REASSESSMENT METHOD AB The past decade has seen the publication of a number of new proposals for the design of phase I trials of anticancer agents. The purpose of these proposals has been to address ethical concerns about treating excessive numbers of patients at subtherapeutic doses of a new agent and to increase the overall efficiency of the process while enhancing the precision of the recommended phase II dose. In early 1998, a workshop of phase I investigators was held under the sponsorship of Bristol-Myers Squibb Pharmaceutical Research Institute (Wallingford, CT) to review the experience to date with novel phase I methodologies, with a particular focus on their efficiency and safety. This report summarizes the material presented. It was concluded that for phase I trials of antineoplastics (cytotoxics), which begin at 0.1 mouse-equivalent LD10 doses, evidence to date suggests that the historic approach of using a modified Fibonacci escalation and three patients per dose level is not necessary and is seldom used. One patient per dose level and more rapid escalation schemes, both empirically based and statistically based, are commonly used with apparent safety. There remain questions, however: Which of the dose escalation schemes is optimal? Are there alternatives to toxicity as a phase I end point, and will these end points be reliable in defining active doses? Answering these questions in a reasonable time frame will be important if new anticancer agents are not to suffer undue delays in phase I evaluation. J Clin Oncol 18:684-692. (C) 2000 by American Society of Clinical Oncology. C1 Queens Univ, Natl Canc Inst Canada, Clin Trials Grp, Kingston, ON K7L 3N6, Canada. Univ Penn, Dept Hematol Oncol, Philadelphia, PA 19104 USA. NCI, Canc Therapy Evaluat Program, Bethesda, MD 20892 USA. Bristol Myers Squibb Pharmaceut Res Inst, Wallingford, CT USA. RP Eisenhauer, EA (reprint author), Queens Univ, Natl Canc Inst Canada, Clin Trials Grp, 82-84 Barrie St, Kingston, ON K7L 3N6, Canada. NR 12 TC 135 Z9 137 U1 1 U2 13 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0732-183X J9 J CLIN ONCOL JI J. Clin. Oncol. PD FEB PY 2000 VL 18 IS 3 BP 684 EP 692 PG 9 WC Oncology SC Oncology GA 281ZX UT WOS:000085192300026 PM 10653884 ER PT J AU Fine, HA Figg, WD Jaeckle, K Wen, PY Kyritsis, AP Loeffler, JS Levin, VA Black, PM Kaplan, R Pluda, JM Yung, WKA AF Fine, HA Figg, WD Jaeckle, K Wen, PY Kyritsis, AP Loeffler, JS Levin, VA Black, PM Kaplan, R Pluda, JM Yung, WKA TI Phase II trial of the antiangiogenic agent thalidomide in patients with recurrent high-grade gliomas SO JOURNAL OF CLINICAL ONCOLOGY LA English DT Article ID ENDOTHELIAL GROWTH-FACTOR; TUMOR ANGIOGENESIS; FACTOR EXPRESSION; INHIBITION; CHEMOTHERAPY; ASTROCYTOMAS; THERAPIES; LYMPHOMA; CRITERIA; DISEASE AB Purpose: Little progress has been made in the treatment of adult high-grade gliomas over the last two decades, thus necessitating a search for novel therapeutic strategies. Malignant gliomas are vascular or angiogenic tumors, which leads to the supposition that angiogenesis inhibition may represent a potentially promising strategy in the treatment of these tumors. We present the results of a phase II trial of thalidomide, a putative inhibitor of angiogenesis, in the treatment of adults with previously irradiated, recurrent high-grade gliomas. Patients and Methods: Patients with a histologic diagnosis of anaplastic mixed glioma, anaplastic astrocytoma, or glioblastoma multiforme who had radiographic demonstration of tumor progression after standard external-beam radiotherapy with or without chemotherapy were eligible. Patients were initially treated with thalidomide 800 mg/d with increases in dose by 200 mg/d every 2 weeks until a final daily dose of 1,200 mg was achieved. Patients were evaluated every 8 weeks for response by both clinical and radiographic criteria. Results: A total of 39 patients were accrued, with 36 patients being assessable for both toxicity and response. Thalidomide was well tolerated, with constipation and sedation being the major toxicities. One patient developed a grade 2 peripheral neuropathy after treatment with thalidomide for nearly a year. There were two objective radiographic partial responses (6%), two minor responses (6%), and 12 patients with stable disease (33%). Eight patients were alive more than 1 year after starting thalidomide, although almost all with tumor progression. Changes in serum levels of basic fibroblastic growth factor (bFGF) were correlated with time to tumor progression and overall survival. Conclusion: Thalidomide is a generally well-tolerated drug that may have antitumor activity in a minority of patients with recurrent high-grade gliomas. Future studies will better define the usefulness of thalidomide in newly diagnosed patients with malignant gliomas and in combination with radiotherapy and chemotherapy. Additionally, studies will be needed to confirm the potential utility of changes in serum bFGF as a marker of antiangiogenic activity and/or glioma growth. (C) 2000 by American Society of Clinical Oncology. C1 Harvard Univ, Sch Med, Brigham & Womens Hosp, Ctr Neurooncol,Dana Farber Canc Inst, Boston, MA 02115 USA. Harvard Univ, Sch Med, Brigham & Womens Hosp, Dept Neurol, Boston, MA 02115 USA. Harvard Univ, Sch Med, Brigham & Womens Hosp, Dept Surg, Boston, MA 02115 USA. Harvard Univ, Sch Med, Brigham & Womens Hosp, Dept Radiat Oncol, Boston, MA 02115 USA. Univ Texas, MD Anderson Canc Ctr, Dept Neurooncol, Houston, TX 77030 USA. Natl Cent Nervous Syst Consortium, Houston, TX USA. NCI, Canc Treatment & Evaluat Program, Investigat Drug Branch, Div Canc Therapy & Diagnosis, Bethesda, MD 20892 USA. RP Fine, HA (reprint author), NINDS, NCI, Neurooncol Branch, NIH, Bldg 10,Rm 12W253,10 Ctr Dr, Bethesda, MD 20892 USA. RI Ain, Kenneth/A-5179-2012; Figg Sr, William/M-2411-2016 OI Ain, Kenneth/0000-0002-2668-934X; NR 34 TC 331 Z9 341 U1 2 U2 7 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0732-183X J9 J CLIN ONCOL JI J. Clin. Oncol. PD FEB PY 2000 VL 18 IS 4 BP 708 EP 715 PG 8 WC Oncology SC Oncology GA 285RL UT WOS:000085401800002 PM 10673511 ER PT J AU Zujewski, J Horak, ID Bol, CJ Woestenborghs, R Bowden, C End, DW Piotrovsky, VK Chiao, J Belly, RT Todd, A Kopp, WC Kohler, DR Chow, C Noone, M Hakim, FT Larkin, G Gress, RE Nussenblatt, RB Kremer, AB Cowan, KH AF Zujewski, J Horak, ID Bol, CJ Woestenborghs, R Bowden, C End, DW Piotrovsky, VK Chiao, J Belly, RT Todd, A Kopp, WC Kohler, DR Chow, C Noone, M Hakim, FT Larkin, G Gress, RE Nussenblatt, RB Kremer, AB Cowan, KH TI Phase I and pharmacokinetic study of farnesyl protein transferase inhibitor R115777 in advanced cancer SO JOURNAL OF CLINICAL ONCOLOGY LA English DT Article ID RAS TRANSGENIC MICE; HUMAN BREAST-CANCER; TUMOR-CELL LINES; K-RAS; ACTIN CYTOSKELETON; N-RAS; MUTATIONS; PRENYLATION; ACTIVATION; PATHWAY AB Purpose: To determine the maximum-tolerated dose, toxicities, and pharmacokinetic profile of the farnesyl protein transferase inhibitor R115777 when administered orally bid for 5 days every 2 weeks. Patients and Methods: Twenty-seven patients with a median age of 58 years received 85 cycles of R115777 using an intrapatient and interpatient dose escalation schema. Drug was administered orally at escalating doses as a solution (25 to 850 mg bid) or as pellet capsules (500 to 1300 mg bid). Pharmacokinetics were assessed after the first dose and the last dose administered during cycle 1. Results: Dose-limiting toxicity of grade 3 neuropathy was observed in one patient and grade 2 fatigue (decrease in two performance status levels) was seen in four of six patients treated with 1.300 mg bid. The most frequent clinical grade 2 or 3 adverse events in any cycle included nausea, vomiting, headache, fatigue, anemia, and hypotension. Myelosuppression was mild and infrequent. Peak plasma concentrations of R115777 were achieved within 0.5 to 4 hours after oral drug administration. The elimination of R115777 from plasma was biphasic, with sequential half-lives of about 5 hours and 16 hours. There was little drug accumulation after bid dosing, and steady-state concentrations were achieved within 2 to 3 days. The pharmacokinetics were dose proportional in the 25 to 325 mg/dose range for the oral solution. Urinary excretion of unchanged R115777 was less than 0.1% of the oral dose. One patient with metastatic colon cancer treated at the 500-mg bid dose had a 46% decrease in carcinoembryonic antigen levels, improvement in cough, and radiographically stable disease for 5 months. Conclusion: R115777 is bioavailable after oral administration and has an acceptable toxicity profile. Based upon pharmacokinetic data, the recommended dose for phase II trials is 500 mg orally bid (total daily dose, 1,000 mg) for 5 consecutive days followed by 9 days of rest. Studies of continuous dosing and studies of R115777 in combination with chemotherapy are ongoing. (C) 2000 by American Society of Clinical Oncology. C1 Natl Canc Inst, Div Clin Sci, Med Branch, Bethesda, MD 20892 USA. Natl Inst Hlth, Ctr Clin, Bethesda, MD USA. NEI, Bethesda, MD 20892 USA. SAIC Frederick, Frederick, MD USA. Janssen Res Inst, Titusville, NJ USA. Janssen Res Fdn, B-2340 Beerse, Belgium. Orthoclin Diagnost, Rochester, NY USA. Johnson & Johnson Res, Sydney, NSW, Australia. RP Zujewski, J (reprint author), Natl Canc Inst, Div Clin Sci, Med Branch, 9000 Rockville Pike, Bethesda, MD 20892 USA. FU NCI NIH HHS [N01-CO-56000] NR 68 TC 204 Z9 210 U1 0 U2 3 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0732-183X J9 J CLIN ONCOL JI J. Clin. Oncol. PD FEB PY 2000 VL 18 IS 4 BP 927 EP 941 PG 15 WC Oncology SC Oncology GA 285RL UT WOS:000085401800027 PM 10673536 ER PT J AU Wieand, S Wolmark, N AF Wieand, S Wolmark, N TI The logic of evidence - In reply SO JOURNAL OF CLINICAL ONCOLOGY LA English DT Letter C1 Natl Surg Adjuvant Breast & Bowel Project, Pittsburgh, PA USA. RP Wieand, S (reprint author), Natl Surg Adjuvant Breast & Bowel Project, Pittsburgh, PA USA. NR 1 TC 0 Z9 0 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0732-183X J9 J CLIN ONCOL JI J. Clin. Oncol. PD FEB PY 2000 VL 18 IS 4 BP 942 EP 942 PG 1 WC Oncology SC Oncology GA 285RL UT WOS:000085401800029 ER PT J AU Zhang, Z Schwartz, S Wagner, L Miller, W AF Zhang, Z Schwartz, S Wagner, L Miller, W TI A greedy algorithm for aligning DNA sequences SO JOURNAL OF COMPUTATIONAL BIOLOGY LA English DT Article DE sequence alignment; greedy algorithms; dynamic programming ID GENERATION; ALIGNMENT; PROGRAM; TOOL AB For aligning DNA sequences that differ only by sequencing errors, or by equivalent errors from other sources, a greedy algorithm can be much faster than traditional dynamic programming approaches and Set produce an alignment that is guaranteed to be theoretically optimal. We introduce a nem greedy alignment algorithm with particularly good performance and show that it computes the same alignment as does a certain dynamic programming algorithm, while executing over 10 times faster on appropriate data. An implementation of this algorithm is currently used in a program that assembles the UniGene database at the National Center for Biotechnology Information. C1 Penn State Univ, Dept Comp Sci & Engn, University Pk, PA 16802 USA. Natl Lib Med, Natl Ctr Biotechnol Informat, NIH, Bethesda, MD 20894 USA. RP Miller, W (reprint author), Penn State Univ, Dept Comp Sci & Engn, University Pk, PA 16802 USA. RI Yang, Chen/G-1379-2010 FU NLM NIH HHS [LM05110] NR 18 TC 1833 Z9 1921 U1 10 U2 101 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1066-5277 J9 J COMPUT BIOL JI J. Comput. Biol. PD FEB-APR PY 2000 VL 7 IS 1-2 BP 203 EP 214 DI 10.1089/10665270050081478 PG 12 WC Biochemical Research Methods; Biotechnology & Applied Microbiology; Computer Science, Interdisciplinary Applications; Mathematical & Computational Biology; Statistics & Probability SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Computer Science; Mathematical & Computational Biology; Mathematics GA 328DJ UT WOS:000087833300012 PM 10890397 ER PT J AU Berman, P Zhang, Z Wolf, YI Koonin, EV Miller, W AF Berman, P Zhang, Z Wolf, YI Koonin, EV Miller, W TI Winnowing sequences from a database search SO JOURNAL OF COMPUTATIONAL BIOLOGY LA English DT Article DE sequence database; sequence alignment; Blast program ID TREES AB In database searches for sequence similarity, matches to a distinct sequence region (e.g., protein domain) are frequently obscured by numerous matches to another region of the same sequence. In order to cope with this problem, algorithms are developed to discard redundant matches, One model for this problem begins with a list of intervals, each with an associated score; each interval gives the range of positions in the query sequence that align to a database sequence, and the score is that of the alignment. If interval I is contained in interval J, and I's score is less than J's, then I is said to be dominated by J, The problem is then to identify each interval that is dominated by at least K other intervals, where K is a given level of "tolerable redundancy." An algorithm is developed to solve the problem in O(N log N) time and O(N*) space, where N is the number of intervals and N* is a precisely defined value that never exceeds N and is frequently much smaller. This criterion for discarding database hits has been implemented in the Blast program, as illustrated herein with examples. Several variations and extensions of this approach are also described. C1 Penn State Univ, Dept Comp Sci & Engn, University Pk, PA 16802 USA. Natl Lib Med, Natl Ctr Biotechnol Informat, NIH, Bethesda, MD 20894 USA. RP Miller, W (reprint author), Penn State Univ, Dept Comp Sci & Engn, University Pk, PA 16802 USA. FU NLM NIH HHS [LM05110] NR 14 TC 5 Z9 5 U1 1 U2 1 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1066-5277 J9 J COMPUT BIOL JI J. Comput. Biol. PD FEB-APR PY 2000 VL 7 IS 1-2 BP 293 EP 302 DI 10.1089/10665270050081531 PG 10 WC Biochemical Research Methods; Biotechnology & Applied Microbiology; Computer Science, Interdisciplinary Applications; Mathematical & Computational Biology; Statistics & Probability SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Computer Science; Mathematical & Computational Biology; Mathematics GA 328DJ UT WOS:000087833300018 PM 10890403 ER PT J AU Blanquet-Grossard, F Sazdovitch, V Jean, A Deslys, JP Dormont, D Hauw, JJ Marion, D Brown, P Cesbron, JY AF Blanquet-Grossard, F Sazdovitch, V Jean, A Deslys, JP Dormont, D Hauw, JJ Marion, D Brown, P Cesbron, JY TI Prion protein is not detectable in dental pulp from patients with Creutzfeldt-Jakob disease SO JOURNAL OF DENTAL RESEARCH LA English DT Letter DE Creutzfeldt-Jacob disease; prion; dentistry; iatrogenic contamination ID SCRAPIE C1 CNRS, IBL, F-59021 Lille, France. HP Pitie Salpetriere, Neuropathol Lab, F-75651 Paris 13, France. Fac Chirurg Dent, Dept Ontol Conservatrice Endodontie, F-44042 Nantes 1, France. CEA, Lab Neurovirol & Neuropathol Expt, F-92265 Fontenay Aux Roses, France. NINDS, Cent Nervous Syst Studies Lab, NIH, Bethesda, MD 20892 USA. RP Cesbron, JY (reprint author), CNRS, IBL, 1 Rue du Pr Calmette,BP447, F-59021 Lille, France. NR 5 TC 21 Z9 23 U1 1 U2 2 PU AMER ASSOC DENTAL RESEARCH PI ALEXANDRIA PA 1619 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0022-0345 J9 J DENT RES JI J. Dent. Res. PD FEB PY 2000 VL 79 IS 2 BP 700 EP 700 PG 1 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA 287FF UT WOS:000085495300001 PM 10728969 ER PT J AU Wang, S Baum, BJ Yamano, S Mankani, MH Sun, D Jonsson, M Davis, C Graham, FL Gauldie, J Atkinson, JC AF Wang, S Baum, BJ Yamano, S Mankani, MH Sun, D Jonsson, M Davis, C Graham, FL Gauldie, J Atkinson, JC TI Adenoviral-mediated gene transfer to mouse salivary glands SO JOURNAL OF DENTAL RESEARCH LA English DT Article DE salivary glands; gene transfer; gene therapy ID IN-VIVO; RECOMBINANT ADENOVIRUSES; IMMUNE-RESPONSES; VECTORS; INTERLEUKIN-10; EXPRESSION; THERAPY; DELIVERY; BARRIERS AB Adenoviral vectors effectively transfer genes to rat salivary glands. However, potent immune responses limit their use in vivo. Mice offer more opportunities than rats for the study of these immune processes. We first established conditions for infection of mouse salivary glands, with an adenoviral vector. The effects of time, viral dose, viral diluent buffer volume, and dexamethasone on expression of a transgene, luciferase, were determined by means of the recombinant vector AdCMVluc. Optimal luciferase expression was observed when the vector was suspended in 50 mu L of buffer. This volume completely filled the gland parenchyma and slightly distended the capsule. Dexamethasone increased immediate transgene expression and reduced the acute inflammation one day following viral administration, but did not alter subsequent mononuclear inflammation or transgene expression 14 or 28 days later. An adenoviral vector encoding either anti-inflammatory cytokine IL-4 or IL-10 was coadministered with AdCMVluc to increase transgene expression at 14 and 28 days. While this strategy did not extend the duration of luciferase expression, co-administration of AdCMVIL-10 with AdCMVluc almost completely eliminated the chronic inflammatory infiltrate in the glands after 28 days. This study demonstrates that adenoviral-mediated gene transfer to mouse submandibular glands is possible by intraductal cannulation and that reduction of either the acute or chronic inflammatory infiltrates was insufficient to increase long-term transgene expression in this tissue. C1 NIDCR, Gene Therapy & Therapeut Branch, NIH, Bethesda, MD 20892 USA. NIDCR, Craniofacial & Skeletal Dis Branch, NIH, Bethesda, MD 20892 USA. McMaster Univ, Hlth Sci Ctr, Dept Biol, Hamilton, ON, Canada. McMaster Univ, Hlth Sci Ctr, Dept Pathol, Hamilton, ON, Canada. RP Atkinson, JC (reprint author), NIDCR, Gene Therapy & Therapeut Branch, NIH, Bldg 10,1N113,MSC-1190, Bethesda, MD 20892 USA. OI Yamano, Seiichi/0000-0003-2056-4359 NR 30 TC 37 Z9 41 U1 1 U2 2 PU AMER ASSOC DENTAL RESEARCH PI ALEXANDRIA PA 1619 DUKE ST, ALEXANDRIA, VA 22314 USA SN 0022-0345 J9 J DENT RES JI J. Dent. Res. PD FEB PY 2000 VL 79 IS 2 BP 701 EP 708 PG 8 WC Dentistry, Oral Surgery & Medicine SC Dentistry, Oral Surgery & Medicine GA 287FF UT WOS:000085495300002 PM 10728970 ER PT J AU Kiesewetter, DO Sassaman, MB Robbins, J Jagoda, EM Carson, RE Appel, NM Sutkowski, E Herscovitch, P Braun, A Eckelman, WC AF Kiesewetter, DO Sassaman, MB Robbins, J Jagoda, EM Carson, RE Appel, NM Sutkowski, E Herscovitch, P Braun, A Eckelman, WC TI Synthesis and evaluation of an F-18 analog of forskolin for imaging adenylyl cyclase SO JOURNAL OF FLUORINE CHEMISTRY LA English DT Article; Proceedings Paper CT 16th International Conference on Fluorine Chemistry (ICFC 99) CY MAY 09-11, 1999 CL TOKYO, JAPAN SP Japan Soc Promot Sci DE adenylyl cyclase; forskolin; F-18 ID 7-DESACETYLFORSKOLIN; DERIVATIVES AB We prepared two fluorine containing carbamate analogs of forskolin as potential agents for imaging adenylyl cyclase in the Living brain with positron emission tomography. Both compounds display high in vitro affinity for adenylyl cyclase. The radiosynthesis of 7-(2-[F-18] fluoroethyl) carbamoyl forskolin from an imidazolyl carbamate utilizes [F-18] fluoroethyl amine, prepared by a novel method employing rapid deprotection and requiring no distillation. The brain uptake in rats and monkeys of 7-(2-[F-18] fluoroethyl) carbamoyl forskolin is too low to be an effective imaging agent for this second messenger system. (C) 2000 Elsevier Science S.A. All rights reserved. C1 NIH, Warren Grant Magnuson Clin Ctr, Posit Emiss Tomog Dept, Bethesda, MD 20892 USA. US FDA, Ctr Biol Evaluat & Res, Mol Pharmacol Lab, Bethesda, MD 20892 USA. Natl Inst Deafness & Other Communicat Disorders, Voice Speech & Language Branch, NIH, Bethesda, MD 20892 USA. RP Kiesewetter, DO (reprint author), NIH, Warren Grant Magnuson Clin Ctr, Posit Emiss Tomog Dept, Bethesda, MD 20892 USA. RI Carson, Richard/H-3250-2011 OI Carson, Richard/0000-0002-9338-7966 NR 11 TC 5 Z9 5 U1 0 U2 1 PU ELSEVIER SCIENCE SA PI LAUSANNE PA PO BOX 564, 1001 LAUSANNE, SWITZERLAND SN 0022-1139 J9 J FLUORINE CHEM JI J. Fluor. Chem. PD FEB PY 2000 VL 101 IS 2 BP 297 EP 304 DI 10.1016/S0022-1139(99)00174-8 PG 8 WC Chemistry, Inorganic & Nuclear; Chemistry, Organic SC Chemistry GA 269QH UT WOS:000084488700025 ER PT J AU Gonzalez, A Kirsch, WG Shirokova, N Pizarro, G Stern, MD Rios, E AF Gonzalez, A Kirsch, WG Shirokova, N Pizarro, G Stern, MD Rios, E TI The spark and its ember - Separately gated local components of Ca2+ release in skeletal muscle SO JOURNAL OF GENERAL PHYSIOLOGY LA English DT Article DE excitation-contraction coupling; calcium sparks; ryanodine receptors; caffeine magnesium ID SARCOPLASMIC-RETICULUM CALCIUM; RAT VENTRICULAR MYOCYTES; CUT TWITCH FIBERS; RYANODINE RECEPTOR; CARDIAC MYOCYTES; CHARGE MOVEMENT; CA-2+ RELEASE; FROG-MUSCLE; CHANNEL; EVENTS AB Amplitude, spatial width, and rise time of Ca2+ sparks were compared in hog fast-twitch muscle, in three conditions that alter activation of release channels by [Ca2+]. A total of similar to 17,000 sparks from 30 cells were evaluated. In cells under voltage clamp, caffeine (0.5 or 1 mM) increased average spark width by 28%, rise time by 18%, and amplitude by 7%. Increases in width were significant even among events of the same rise time. Spontaneous events recorded in permeabilized fibers with low internal [Mg2+] (0.4 mM), had width and rise times greater than in reference, and not significantly different than those in caffeine. The spark average in reference rides on a continuous fluorescence "ridge" and is continued by an "ember," a prolongation of width similar to 1 mu m and amplitude <0.2, vanishing in similar to 100 ms. Ridge and ember were absent in caffeine and in permeabilized cells. Ex posure of voltage-clamped cells to high internal [Mg2+] (7 mM) had effects opposite to caffeine, reducing spark width by 26% and amplitude by 27%. In high [Mg2+], the ember was visible in individual sparks as a prolongation of variable duration and amplitude up to 1.2. Based on simulations and calculation of Ca2+ release flux from averaged sparks, the increase in spark width caused by caffeine was interpreted as evidence of an increase in radius of the release source-presumably by recruitment of additional channels. Conversely, spark narrowing suggests loss of contributing channels in high Mg2+. Therefore, these changes in spark width at constant rise times are evidence of a multichannel origin of sparks. Because ridge and ember were reduced by promoters of Ca2+-dependent activation (caffeine, low [Mg2+]) and became more visible in the presence of its inhibitors, they are probably manifestations of Ca2+ release directly operated by voltage sensors. C1 Rush Univ, Dept Mol Biophys & Physiol, Chicago, IL 60612 USA. Univ Republ, Fac Med, Dept Biofis, Montevideo, Uruguay. NIA, Cardiovasc Sci Lab, NIH, Baltimore, MD 21224 USA. RP Gonzalez, A (reprint author), Rush Univ, Dept Mol Biophys & Physiol, 1750 W Harrison St,Suite 1279 JS, Chicago, IL 60612 USA. NR 67 TC 62 Z9 63 U1 0 U2 0 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0022-1295 J9 J GEN PHYSIOL JI J. Gen. Physiol. PD FEB PY 2000 VL 115 IS 2 BP 139 EP 157 DI 10.1085/jgp.115.2.139 PG 19 WC Physiology SC Physiology GA 286PV UT WOS:000085455100004 PM 10653893 ER PT J AU Jensen, KT Wolfinbarger, JB Aasted, B Bloom, ME AF Jensen, KT Wolfinbarger, JB Aasted, B Bloom, ME TI Replication of Aleutian mink disease parvovirus in mink lymph node histocultures SO JOURNAL OF GENERAL VIROLOGY LA English DT Article ID FOLLICULAR DENDRITIC CELLS; CENTER B-CELLS; MONOCLONAL-ANTIBODIES; ADULT MINK; INTERSTITIAL PNEUMONIA; NUCLEOTIDE-SEQUENCE; VIRAL REPLICATION; INFECTION; VIRUS; MACROPHAGES AB Aleutian mink disease parvovirus (ADV), causes an immune disorder with a persistent infection of lymphoid organs in adult mink. We studied replication of ADV in gel-supported histocultures prepared from adult mink mesenteric lymph node (MLN). Evidence of virus replication in the histocultures was first observed by indirect immunofluorescence 72 h after incubation with virus. Cells resembling lymphocytes and macrophages contained both ADV capsid (VP2) and nonstructural (NS1 and NS2) proteins, and were present in a distribution suggestive of infected cells within germinal centres. ADV replicative form and encapsidated virion DNA were also detected in infected histocultures at time-points after 72 h. In addition, we were able to passage ADV-Utah to a new round of histocultures. These results suggested that the infected cells were actual target cells for ADV replication and that productive ADV-Utah replication, complete with the generation of virus, was occurring in the histocultures. The mink MLN histocultures provide a system to study the replication and molecular pathogenesis of ADV in target tissues. C1 NIAID, Rocky Mt Labs, Persistent Viral Dis Lab, NIH, Hamilton, MT 59840 USA. Royal Vet & Agr Univ, Inst Vet Microbiol, Lab Virol & Immunol, DK-1870 Frederiksberg C, Denmark. RP Bloom, ME (reprint author), NIAID, Rocky Mt Labs, Persistent Viral Dis Lab, NIH, Hamilton, MT 59840 USA. NR 46 TC 4 Z9 4 U1 0 U2 5 PU SOC GENERAL MICROBIOLOGY PI READING PA MARLBOROUGH HOUSE, BASINGSTOKE RD, SPENCERS WOODS, READING, BERKS, ENGLAND RG7 1AE SN 0022-1317 J9 J GEN VIROL JI J. Gen. Virol. PD FEB PY 2000 VL 81 BP 335 EP 343 PN 2 PG 9 WC Biotechnology & Applied Microbiology; Virology SC Biotechnology & Applied Microbiology; Virology GA 279DF UT WOS:000085028300004 PM 10644831 ER PT J AU Dressman, MA Olivos-Glander, IM Nussbaum, RL Suchy, SF AF Dressman, MA Olivos-Glander, IM Nussbaum, RL Suchy, SF TI Ocrl1, a Ptdlns(4.5)P-2 5-phosphatase, is localized to the trans-Golgi network of fibroblasts and epithelial cells SO JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY LA English DT Article DE Golgi apparatus; Lowe syndrome; phosphatidylinositol 4,5-bisphosphate; 5-phosphatase; renal proximal tubules; trans-Golgi network ID MANNOSE 6-PHOSPHATE RECEPTOR; PHOSPHOLIPASE-C ISOZYMES; BREFELDIN-A; LOWE-SYNDROME; PHOSPHATIDYLINOSITOL 4,5-BISPHOSPHATE; SIGNAL-TRANSDUCTION; COATED VESICLES; EARLY ENDOSOMES; MEMBRANE; LYSOSOMES AB Ptdlns(4,5)P-2 and Ptdlns(4,5)P-2 5-phosphatases play important roles in diverse aspects of cell metabolism, including protein trafficking. However, the relative importance of the Ptdlns(4,5)P-2 5-phosphatases in regulating Ptdlns(4,5)P-2 levels for specific cell processes is not well understood. Ocrl1 is a Ptdlns(4,5)P-2 5-phosphatase that is deficient in the oculocerebrorenal syndrome of Lowe, a disorder characterized by defects in kidney and lens epithelial cells and mental retardation. Ocrl1 was originally localized to the Golgi in fibroblasts, but a subsequent report suggested a lysosomal localization in a kidney epithelial cell line. In this study we defined the localization of ocrl1 in fibroblasts and in two kidney epithelial cell lines by three methods: immunofluorescence, subcellular fractionation, and a dynamic perturbation assay with brefeldin A. We found that ocrl1 was a Golgi-localized protein in all three cell types and further identified it as a protein of the trans-Golgi network (TGN). The TGN is a major sorting site and has the specialized function in epithelial cells of directing proteins to the apical or basolateral domains. The epithelial cell phenotype in Lowe syndrome and the localization of ocrl1 to the TGN imply that this Ptdlns(4,5)P-2 5-phosphatase plays a role in trafficking. C1 NIH, Natl Human Genome Res Inst, Genet Dis Res Branch, Bethesda, MD 20892 USA. RP Suchy, SF (reprint author), NIH, Natl Human Genome Res Inst, Genet Dis Res Branch, 49 Convent Dr,Room 4A66, Bethesda, MD 20892 USA. NR 50 TC 74 Z9 74 U1 0 U2 1 PU HISTOCHEMICAL SOC INC PI SEATTLE PA UNIV WASHINGTON, DEPT BIOSTRUCTURE, BOX 357420, SEATTLE, WA 98195 USA SN 0022-1554 J9 J HISTOCHEM CYTOCHEM JI J. Histochem. Cytochem. PD FEB PY 2000 VL 48 IS 2 BP 179 EP 189 PG 11 WC Cell Biology SC Cell Biology GA 288AV UT WOS:000085540400003 PM 10639484 ER PT J AU Bagrov, AY Dmitrieva, RI Dorofeeva, NA Fedorova, OV Lopatin, DA Lakatta, EG Droy-Lefaix, MT AF Bagrov, AY Dmitrieva, RI Dorofeeva, NA Fedorova, OV Lopatin, DA Lakatta, EG Droy-Lefaix, MT TI Cicletanine reverses vasoconstriction induced by the endogenous sodium pump ligand, marinobufagenin, via a protein kinase C dependent mechanism SO JOURNAL OF HYPERTENSION LA English DT Article DE Na/K-ATPase; marinobufagenin; protein kinase C; cicletanine; hypertension ID RAT AORTA; SMOOTH-MUSCLE; ALPHA-SUBUNIT; HUMAN PLASMA; INHIBITION; OUABAIN; ATPASE; CELLS; PHOSPHODIESTERASES; PHOSPHORYLATION AB Rationale Cicletanine (CIC), an anti-hypertensive compound with direct vascular and natriuretic actions, is especially effective in salt-sensitive hypertension, in which dysregulation of the sodium pump plays an important pathogenic role, and digitalis-like cardiotonic steroids contribute to increased vascular tone. The purpose of the present study was to investigate whether, and by what mechanisms, cicletanine antagonizes the vasoconstrictor effects of cardiotonic steroids in isolated human arteries. Methods The effects of cicletanine on vascular tone were studied in isolated, endothelium-denuded rings of 2nd-3rd-order branches of human mesenteric arteries precontracted with bufodienolide marinobufagenin (MBG), an Na/K-ATPase inhibitor, or endothelin-1 (ET-1). Na/K-ATPase activity was measured in sarcolemmal membranes from the mesenteric artery. Activity of rat brain protein kinase C (PKC) was measured using the PepTag phosphorylation assay. Results MBG and ET-1 both induced sustained vasoconstriction in human mesenteric artery rings, and cicletanine relaxed rings pre-contracted with either MBG (EC50 = 11 +/- 2 mu mol/l) or ET-1 (EC50 = 6.4 +/- 1.1 mu mol/l). Although 8-Br-cGMP (100 mu mol/l) caused complete vasorelaxation of arterial rings pre-contracted with ET-1, it did not affect the MBG-induced vasoconstriction. An activator of PKC, phorbol diacetate (PDA) (50 nmol/l), attenuated CIC-induced vasorelaxation of mesenteric artery rings pre-contracted with MBG (EC50 >100 mu mol/l), but not rings pre-contracted with ET-1 (EC50 = 6.5 +/- 1.2 mu mol/l), In mesenteric artery sarcolemma, 100 nmol/l MBG inhibited the Na/K-ATPase by 68 +/- 5% and cicletanine (100 mu mol/l) attenuated this Na/K-ATPase inhibition by 85 +/- 6%. In the PepTag PKC assay, cicletanine produced a concentration-dependent inhibition of rat brain PKC activity (IC50 45 +/- 11 mu mol/l). In the presence of 50 nmol/l PDA, 100 mu mol/l cicletanine did not antagonize the Na/K-ATPase inhibition by MBG, and did not inhibit the PKC from rat brain. Conclusions Cicletanine antagonizes vasoconstriction induced by Na/K-ATPase inhibition via a PKC-dependent mechanism that does not involve inhibition of cyclic GMP phosphodiesterase (cGMP-PDE), This mechanism of action may be relevant to the greater potency of cicletanine in salt-sensitive hypertension in which plasma levels of endogenous digitalis-like cardiotonic steroids are elevated. Our findings also suggest that PKC is an important factor for cardiotonic steroid - Na/K-ATPase interactions on the vascular tone, and is therefore a potential target for therapeutic intervention in hypertension. J Hypertens 2000, 18:209-215 (C) Lippincott Williams & Wilkins. C1 NIA, Cardiovasc Sci Lab, Intramural Res Program, Baltimore, MD 21224 USA. IM Sechenov Evolutionary Physiol & Biochem Inst, Pharmacol Lab, St Petersburg 194223, Russia. IPSEN Inst, Paris, France. RP Bagrov, AY (reprint author), NIA, Cardiovasc Sci Lab, Intramural Res Program, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. OI Dmitrieva, Renata/0000-0002-3073-7914 NR 37 TC 10 Z9 11 U1 1 U2 3 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0263-6352 J9 J HYPERTENS JI J. Hypertens. PD FEB PY 2000 VL 18 IS 2 BP 209 EP 215 DI 10.1097/00004872-200018020-00012 PG 7 WC Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 283CH UT WOS:000085258200012 PM 10694190 ER PT J AU Ohta, K Wiggert, B Yamagami, S Taylor, AW Streilein, JW AF Ohta, K Wiggert, B Yamagami, S Taylor, AW Streilein, JW TI Analysis of immunomodulatory activities of aqueous humor from eyes of mice with experimental autoimmune uveitis SO JOURNAL OF IMMUNOLOGY LA English DT Article ID GROWTH-FACTOR-BETA; ENDOTOXIN-INDUCED UVEITIS; RETINOID-BINDING PROTEIN; TUMOR-NECROSIS-FACTOR; CILIARY BODY CELLS; ANTERIOR-CHAMBER; IMMUNE PRIVILEGE; T-CELLS; IL-6-DEFICIENT MICE; INTERLEUKIN-6 IL-6 AB Aqueous humor (AqH) contains immunosuppressive factors, especially TGF-beta 2, that contribute to the immune privileged status of the anterior chamber, However, this may not be true when the blood-ocular barrier is compromised by ocular inflammation. To determine the immunosuppressive status of AqH from murine eyes afflicted with experimental autoimmune uveitis, B10.A mice were immunized with interphotoreceptor retinoid-binding protein. AqH was collected from eyes of affected mice periodically after immunization and then evaluated for content of TGF-beta, proinflammatory cytokines, and the capacity to suppress anti-CD3-driven T cell proliferation. mRNA expression of selected cytokines in iris and ciliary body from inflamed eyes was analyzed by ribonuclease protection assay. We found that TGF-beta levels were significantly increased in AqH from EAU eyes on days 11, 17, and 28, AqH collected on day 11 (onset of disease) failed to suppress T cell proliferation and contained large amounts of locally produced IL-6 that antagonized TGF-beta, In contrast, AqH collected at 17 days (when ocular inflammation was progressively severe) reexpressed the ability to suppress T cell proliferation, in this case due to high levels of blood-derived TGF-beta 1 and eye-derived TGF-beta 2 in the absence of IL-6. Thus, during the onset of experimental autoimmune uveitis, the ocular microenvironment loses its immunosuppressive properties due to local production of IL-6. But as inflammation mounts, AqH IL-6 content falls, and the fluid reacquires sufficient TGF-beta eventually to suppress immunogenic inflammation. The paradoxical roles of IL-6 in antagonizing TGF-beta, while promoting TGF-beta accumulation during ocular inflammation, is discussed. C1 Schepens Eye Res Inst, Boston, MA 02114 USA. Harvard Univ, Sch Med, Dept Ophthalmol, Boston, MA 02114 USA. NIH, Lab Retinal & Mol Biol, Bethesda, MD 20892 USA. RP Streilein, JW (reprint author), Schepens Eye Res Inst, 20 Staniford St, Boston, MA 02114 USA. FU NEI NIH HHS [EY05678] NR 48 TC 52 Z9 54 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 FEB 1 PY 2000 VL 164 IS 3 BP 1185 EP 1192 PG 8 WC Immunology SC Immunology GA 277AJ UT WOS:000084910300007 PM 10640729 ER PT J AU Huang, H Paul, WE AF Huang, H Paul, WE TI Protein tyrosine phosphatase activity is required for IL-4 induction of IL-4 receptor alpha-chain SO JOURNAL OF IMMUNOLOGY LA English DT Article ID JAK-3 JANUS KINASE; FACTOR-I RECEPTOR; GAMMA-CHAIN; BETA-CHAIN; MYELOID CELLS; INTERLEUKIN-4; PHOSPHORYLATION; SHP-1; EXPRESSION; ACTIVATION AB To investigate the role of protein tyrosine phosphatases in IL-4R alpha-chain expression and signaling, we first established that SHP-1, but not SHP-2, coimmunoprecipitated with anti-IL-4R alpha chain Abs in extracts prepared from resting lymphocytes, We further observed that the protein tyrosine phosphatase inhibitors Na3VO4 and pervanadate blocked the striking induction of IL-4R alpha-chain expression that is mediated by IL-4. However, Na3VO4 did not diminish IL-4-induced Stat6 phosphorylation nor did it block the IL-4-mediated increase in IL-4R alpha-chain mRNA, The striking inhibition in total cellular IL-4R alpha-chain and in cell surface IL-4 receptors was associated with an inhibition of biosynthetic labeling of IL-4R alpha-chain after a 30- min pulse with [S-35] methionine, indicating that reduction of IL-4R alpha-chain protein resulted from either a diminished production of the receptor or a rapid degradation, possibly as a result of phosphorylation of the receptor in an early biosynthetic cellular compartment. Control of newly synthesized IL-4R alpha-chain protein expression by phosphatase may provide a novel means to regulate IL-4 responsiveness. C1 NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Paul, WE (reprint author), NIAID, Immunol Lab, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 38 TC 26 Z9 27 U1 1 U2 1 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD FEB 1 PY 2000 VL 164 IS 3 BP 1211 EP 1215 PG 5 WC Immunology SC Immunology GA 277AJ UT WOS:000084910300010 PM 10640732 ER PT J AU Yu, CR Peden, KWC Zaitseva, MB Golding, H Farber, JM AF Yu, CR Peden, KWC Zaitseva, MB Golding, H Farber, JM TI CCR9A and CCR9B: Two receptors for the chemokine CCL25/TECK/Ck beta-15 that differ in their sensitivities to ligand SO JOURNAL OF IMMUNOLOGY LA English DT Article ID PROTEIN-COUPLED RECEPTOR; IMMUNODEFICIENCY-VIRUS TYPE-1; MOLECULAR-CLONING; FUNCTIONAL-CHARACTERIZATION; CELL-LINES; IN-VIVO; EXPRESSION; LYMPHOCYTES; CXCR4; GENE AB We isolated cDNAs for a chemokine receptor-related protein having the database designation GPR-9-6. Two classes of cDNAs were identified from mRNAs that arose by alternative splicing and that encode receptors that we refer to as CCR9A and CCR9B, CCR9A is predicted to contain 12 additional amino acids at its N terminus as compared with CCR9B, Cells transfected with cDNAs for CCR9A and CCR9B responded to the chemokine CC chemokine ligand 25 (CCL25)/thymus-expressed chemokine (TECK)/chemokine beta-15 (CK beta-15) in assays for both calcium flux and chemotaxis. No other chemokines tested produced responses specific for the cDNA-transfected cells. mRNA for CCR9A/B is expressed predominantly in the thymus, coincident with the expression of CCL25, and highest expression for CCR9A/B among thymocyte subsets was found in CD4(+)CD8(+) cells. mRNAs encoding the A and B forms of the receptor were expressed at a ratio of similar to 10:1 in immortalized T cell lines, in PBMC, and in diverse populations of thymocytes, The EC50 of CCL25 for CCR9A was lower than that for CCR9B, and CCR9A was desensitized by doses of CCL25 that failed to silence CCR9B, CCR9 is the first example of a chemokine receptor in which alternative mRNA splicing leads to proteins of differing activities, providing a mechanism for extending the range of concentrations over which a cell can respond to increments in the concentration of ligand, The study of CCR9A and CCR9B should enhance our understanding of the role of the chemokine system in T cell biology, particularly during the stages of thymocyte development. C1 NIAID, Clin Invest Lab, NIH, Bethesda, MD 20892 USA. US FDA, Ctr Biol Evaluat & Res, Lab Retrovirus Res, Bethesda, MD 20892 USA. RP Farber, JM (reprint author), NIAID, Clin Invest Lab, NIH, Bldg 10,Room 11N-228,MSC 1888, Bethesda, MD 20892 USA. NR 58 TC 39 Z9 40 U1 0 U2 4 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD FEB 1 PY 2000 VL 164 IS 3 BP 1293 EP 1305 PG 13 WC Immunology SC Immunology GA 277AJ UT WOS:000084910300021 PM 10640743 ER PT J AU Gonsky, R Deem, RL Bream, JH Lee, DH Young, HA Targan, SR AF Gonsky, R Deem, RL Bream, JH Lee, DH Young, HA Targan, SR TI Mucosa-specific targets for regulation of IFN-gamma expression: Lamina propria T cells use different cis-elements than peripheral blood T cells to regulate transactivation of IFN-gamma expression SO JOURNAL OF IMMUNOLOGY LA English DT Article ID INTERLEUKIN-2 GENE-EXPRESSION; MESSENGER-RNA STABILITY; ACCESSORY MOLECULE CD28; INTERFERON-GAMMA; TRANSCRIPTIONAL REGULATION; ACTIVATION PATHWAY; C-JUN; PROMOTER; MECHANISMS; LYMPHOCYTES AB Activation of lamina propria (LP)T cells via the CD2 pathway enhances IFN-gamma (IFN-gamma) secretion with further enhancement after CD28 coligation, The molecular mechanisms regulating IFN-gamma expression in LP T cells remain unknown. Previous studies in PBL and T cell lines identified cis- and trans-regulatory elements in TCR-mediated expression of IFN-gamma, This study examines CD2 and PMA/ionophore-responsive IFN-gamma promoter elements, Activation of LPMC via CD2-induced IFN-gamma secretion and a parallel up-regulation of mRNA expression. CD28 coligation enhanced mRNA stability without up-regulating transcription as measured by nuclear run-on. Transfection of a --2.7-kb IFN-gamma promoter-reporter construct into PBL and LP mononuclear cells (LPMC) revealed significant promoter activity after CD2 activation, with additional transactivation after CD2/CD28 costimulation in PBL, but not in LPMC. Functional analysis using truncated promoter fragments identified distinct cis-regulatory regions selectively transactivating LFN-gamma expression in PBL compared with LPMC. In PBL, CD2 activation elements reside within the -108- to +64-bp region, However, in LPMC the upstream region between --204 and --108 bp was essential. Transfection of the proximal and distal AP-l-binding elements, as well as TRE/AP-1 constructs, revealed functional activation of AP-1 subsequent to CD2 signaling, with activation critical in PBL but diminished in LPMC. Electromobility shift analysis using oligonucleotides encompassing the proximal, distal, and BED/AP-1-hinding regions failed to demonstrate selective transactivation after CD2 signaling of LPMC, This report provides evidence that activation of LPMC results in transactivation of multiple promoter elements regulating LFN-gamma expression distinct from those in PBL. C1 Cedars Sinai Med Ctr, Inflammatory Bowel Dis Res Ctr, Los Angeles, CA 90048 USA. NCI, Frederick Canc Res & Dev Ctr, Expt Immunol Lab, Frederick, MD 21702 USA. Seoul Surg Clin, Seoul, South Korea. RP Targan, SR (reprint author), Cedars Sinai Med Ctr, Inflammatory Bowel Dis Res Ctr, 8700 Beverly Blvd,D4063, Los Angeles, CA 90048 USA. RI Young, Howard/A-6350-2008 OI Young, Howard/0000-0002-3118-5111 FU NIDDK NIH HHS [DK-46763, DK-43211] NR 47 TC 19 Z9 19 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 FEB 1 PY 2000 VL 164 IS 3 BP 1399 EP 1407 PG 9 WC Immunology SC Immunology GA 277AJ UT WOS:000084910300033 PM 10640755 ER PT J AU Tamura, M Milenic, DE Iwahashi, M Padlan, E Schlom, J Kashmiri, SVS AF Tamura, M Milenic, DE Iwahashi, M Padlan, E Schlom, J Kashmiri, SVS TI Structural correlates of an anticarcinoma antibody: Identification of specificity-determining residues (SDRs) and development of a minimally immunogenic antibody variant by retention of SDRs only SO JOURNAL OF IMMUNOLOGY LA English DT Article ID TUMOR-ASSOCIATED GLYCOPROTEIN-72; 2ND-GENERATION MONOCLONAL-ANTIBODIES; METASTATIC COLON-CARCINOMA; RADIOIMMUNOTHERAPY TRIAL; PHASE-I; INTRAPERITONEAL RADIOIMMUNOTHERAPY; COLORECTAL-CANCER; TAG-72 EXPRESSION; ALPHA-INTERFERON; OVARIAN-CANCER AB Clinical utility of murine mAbs is limited because many elicit Abs to murine Ig constant and variable regions in patients. An Ab humanized by the current procedure of grafting all the complementarity determining regions (CDRs) of a murine Ab onto the human Ab frameworks is likely to be less immunogenic, except that its murine CDRs could still evoke an anti-variable region response, Previous studies with anticarcinoma mAb CC49 showed that light chain LCDR1 and LCDR2 of humanized CC49 could be replaced with the corresponding CDRs of a human Ab with minimal loss of Ag-binding activity, The studies reported in this paper were undertaken to dissect the CC49 Ag-binding site to identify 1) specificity determining residues (SDRs), the residues of the hypervariable region that are most critical in Ag-Ab interaction, and 2) those residues that contribute to the Idiotopes that are potential targets of patients' immune responses. A panel of variants generated by genetic manipulation of the murine CC49 hypervariable regions were evaluated for their relative Ag-binding affinity and reactivity to sera from several patients who had been immunized with murine CC49. One variant, designated HuCC49V10, retained only the SDRs of CC49 and does not react with the anti-variable region Abs of the sera from the murine CC49-treated patients, These studies thus demonstrate that the genetic manipulation of Ab variable regions can be accomplished by grafting only the SDRs of a xenogeneic Ab onto human Ab frameworks. This approach may reduce the immunogenicity of Abs to a minimum. C1 NCI, NIH, Tumor Immunol & Biol Lab, Bethesda, MD 20892 USA. NCI, NIDDK, Mol Biol Lab, Bethesda, MD 20892 USA. RP Schlom, J (reprint author), NCI, NIH, Tumor Immunol & Biol Lab, Bldg 10,Room 8B07, Bethesda, MD 20892 USA. NR 56 TC 36 Z9 38 U1 0 U2 1 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD FEB 1 PY 2000 VL 164 IS 3 BP 1432 EP 1441 PG 10 WC Immunology SC Immunology GA 277AJ UT WOS:000084910300037 PM 10640759 ER PT J AU Xie, ZH Zhang, J Siraganian, RP AF Xie, ZH Zhang, J Siraganian, RP TI Positive regulation of c-Jun N-terminal kinase and TNF-alpha production but not histamine release by SHP-1 in RBL-2H3 mast cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID PROTEIN-TYROSINE-PHOSPHATASE; AFFINITY IGE RECEPTOR; FC-EPSILON-RI; COMMON ANTIGEN CD45; SIGNAL-TRANSDUCTION; IMMUNOGLOBULIN-E; MEDIATED DEGRANULATION; PHOSPHORYLATION; ACTIVATION; SYK AB The SH2-containing protein tyrosine phosphatase(1) (SHP-1) is important for signaling from immune receptors, To investigate the role of SHP-1 in mast cells we overexpressed the wild-type and the phosphatase-inactive forms of SHP-1 in rat basophilic leukemia 2H3 (RBL-2H3) mast cell line. The phosphatase-inactive SHP-1 (C453S or D419A) retains its ability to bind tyrosine phosphorylated substrates and thereby competes with the endogenous wild-type enzyme. Overexpression of wild-type SHP-1 decreased the Fc epsilon RI aggregation-induced tyrosine phosphorylation of the beta and gamma subunits of the receptor whereas the dominant negative SHP-1 enhanced phosphorylation. There were also similar changes in the tyrosine phosphorylation of Syk. However, receptor-induced histamine release in the cells expressing either wild-type or dominant negative SHP-1 was similar to that in the parental control cells. In contrast, compared with the parental RBL-2H3 cells, Fc epsilon RI-induced c-Jun N-terminal kinase phosphorylation and the level of TNF-alpha mRNA was increased in the cells overexpressing wild-type SHP-1 whereas the dominant negative SHP-1 had the opposite effect. The substrate-trapping mutant SHP1/D419A identified pp25 and pp30 as two major potential substrates of SHP-1 in RBL-2H3 cells. Therefore, SHP-1 may play a role in allergy and inflammation by regulating mast cell cytokine production. C1 Natl Inst Dent & Craniofacial Res, Receptors & Signal Transduct Sect, OIIB, NIH, Bethesda, MD 20892 USA. RP Xie, ZH (reprint author), Natl Inst Dent & Craniofacial Res, Receptors & Signal Transduct Sect, OIIB, NIH, Bldg 10-1A26, Bethesda, MD 20892 USA. NR 56 TC 49 Z9 50 U1 0 U2 1 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD FEB 1 PY 2000 VL 164 IS 3 BP 1521 EP 1528 PG 8 WC Immunology SC Immunology GA 277AJ UT WOS:000084910300048 PM 10640770 ER PT J AU Mathys, JM Melanson, SM Schiffer-Alberts, DJ Ioannidis, JPA Koziel, H Skolnik, PR AF Mathys, JM Melanson, SM Schiffer-Alberts, DJ Ioannidis, JPA Koziel, H Skolnik, PR TI NF-kappa B modulates TNF-alpha production by alveolar macrophages in asymptomatic HIV-seropositive individuals SO JOURNAL OF IMMUNOLOGY LA English DT Article ID TUMOR-NECROSIS-FACTOR; PULMONARY INFECTIOUS COMPLICATIONS; IMMUNODEFICIENCY-VIRUS INFECTION; PROTEASE INHIBITORS; ACTIVATION; CELLS; BETA; PHOSPHORYLATION; DISSOCIATION; EXPRESSION AB Local TNF-alpha production in different organs may affect HIV replication and pathogenesis. Alveolar macrophages (AMs) obtained by bronchoalveolar lavage from asymptomatic HIV-seropositive and HIV-seronegative individuals did not spontaneously release TNF-alpha, but LPS stimulation of these cells significantly increased TNF-alpha production. We tested whether NF-kappa B affects TNF-cy production by AMs using N-tosyl-L-phenylalanine chloromethylketone (TPCK) or N-benzoyl-L-tyrosine ethyl ester (BTEE), which inhibit the degradation of I kappa B, or tricyclodecan-9-yl-xanthogenate-potassium (D609), which inhibits phospholipase C. Alveolar macrophages were exposed to LPS alone and with the chemical protease inhibitors TPCK, BTEE, and D609, NF-kappa B DNA binding induced by LPS treatment of AMs was inhibited by TPCK, BTEE, and D609, These agents also inhibited TNF-alpha mRNA and TNF-alpha protein production. After 24 h, the levels of TNF-alpha mRNA reached equilibrium, as assessed by RT-PCR, The levels of NF-kappa B mRNA remained constant under all conditions. The levels of I kappa B-alpha mRNA Here similar after 30, 60, and 180 min, but the I kappa B-beta mRNA concentration was initially low and increased over time under all conditions, I kappa B-alpha and I kappa B-alpha protein production was not affected by the chemical protease inhibitors. Our data show that TNF-alpha production by LPS-stimulated AMs from asymptomatic HIV-seropositive and -seronegative individuals is regulated via the phospholipase C pathway and by NF-kappa B DNA binding activity without obvious changes in I kappa B-alpha or I kappa B-beta protein concentrations. C1 Tufts Univ, New England Med Ctr, Tupper Res Inst, Dept Med,Div Geog Med & Infect Dis, Boston, MA 02111 USA. NIH, HIV Res Branch, Div Aids, Bethesda, MD 20892 USA. Beth Israel Deaconess Med Ctr, Dept Med, Div Pulm & Crit Care Med, Boston, MA 02215 USA. Harvard Med Sch, Boston, MA 02215 USA. RP Skolnik, PR (reprint author), Tufts Univ, New England Med Ctr, Tupper Res Inst, Dept Med,Div Geog Med & Infect Dis, 750 Washington St, Boston, MA 02111 USA. RI Ioannidis, John/G-9836-2011 FU NCRR NIH HHS [MO1-RR00054]; NHLBI NIH HHS [R01HL57875]; NIAID NIH HHS [P30AI42853] NR 37 TC 8 Z9 8 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 FEB 1 PY 2000 VL 164 IS 3 BP 1588 EP 1594 PG 7 WC Immunology SC Immunology GA 277AJ UT WOS:000084910300057 PM 10640779 ER PT J AU Hartmann, G Weeratna, RD Ballas, ZK Payette, P Blackwell, S Suparto, I Rasmussen, WL Waldschmidt, M Sajuthi, D Purcell, RH Davis, HL Krieg, AM AF Hartmann, G Weeratna, RD Ballas, ZK Payette, P Blackwell, S Suparto, I Rasmussen, WL Waldschmidt, M Sajuthi, D Purcell, RH Davis, HL Krieg, AM TI Delineation of a CpG phosphorothioate oligodeoxynucleotide for activating primate immune responses in vitro and in vivo SO JOURNAL OF IMMUNOLOGY LA English DT Article ID IFN-GAMMA PRODUCTION; BACTERIAL-DNA; IN-VIVO; B-CELL; IMMUNOSTIMULATORY OLIGODEOXYNUCLEOTIDES; ANTISENSE OLIGONUCLEOTIDES; DENDRITIC CELLS; CUTTING EDGE; MURINE; MOTIFS AB Oligodeoxynucleotides (ODN) containing unmethylated CpG dinucleotides within specific sequence contexts (CpG motifs) are detected, like bacterial or viral DNA, as a danger signal by the vertebrate immune system, CpG ODN synthesized with a nuclease-resistant phosphorothioate backbone have been shown to be potent Th1-directed adjuvants in mice, but these motifs have been relatively inactive on primate leukocytes in vitro, Moreover, in vitro assays that predict in vivo adjuvant activity for primates have not been reported. In the present study we tested a panel of CPG ODN for their in vitro and in vivo immune effects in mice and identified in vitro activation of B and NK cells as excellent predictors of in vivo adjuvant activity. Therefore, we tested >250 phosphorothioate ODN for their capacity to stimulate proliferation and CD86 expression of human B cells and to induce lytic activity and CD69 expression of human NK cells. These studies revealed that the sequence, number, and spacing of individual CpG motifs contribute to the immunostimulatory activity of a CpG phosphorothioate ODN, An ODN with a TpC dinucleotide at the 5' end followed by three 6 mer CpG motifs (5'-GTCGTT-3') separated by TpT dinucleotides consistently showed the highest activity for human, chimpanzee, and rhesus monkey leukocytes. Chimpanzees or monkeys vaccinated once against hepatitis B with this CpG ODN adjuvant developed 15 times higher anti-hepatitis B Ab titers than those receiving vaccine alone. In conclusion, we report an optimal human CpG motif for phosphorothioate ODN that is a candidate human vaccine adjuvant. C1 Univ Iowa, Dept Internal Med, Iowa City, IA 52242 USA. CpG ImmunPharmaceut GmbH, Hilden, Germany. Ottawa Hosp, Loeb Hlth Res Inst, Ottawa, ON, Canada. Univ Ottawa, Fac Hlth Sci, Ottawa, ON, Canada. Univ Ottawa, Fac Med, Ottawa, ON, Canada. Vet Affairs Med Ctr, Iowa City, IA 52246 USA. Bogor Agr Univ, Primate Res Ctr, Bogor, Indonesia. NIAID, Hepatitis Viruses Sect, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. CpG Immunopharmaceut Inc, Wellesley, MA 02481 USA. RP Krieg, AM (reprint author), Univ Iowa, Dept Internal Med, 540 EMRB, Iowa City, IA 52242 USA. FU NCI NIH HHS [P01-CA66570]; NIDDK NIH HHS [DK25295] NR 44 TC 445 Z9 475 U1 0 U2 9 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD FEB 1 PY 2000 VL 164 IS 3 BP 1617 EP 1624 PG 8 WC Immunology SC Immunology GA 277AJ UT WOS:000084910300061 PM 10640783 ER PT J AU Wang, ZH Kjellberg, L Abdalla, H Wiklund, F Eklund, C Knekt, P Lehtinen, M Kallings, I Lenner, P Hallmans, G Mahlck, CG Wadell, G Schiller, J Dillner, J AF Wang, ZH Kjellberg, L Abdalla, H Wiklund, F Eklund, C Knekt, P Lehtinen, M Kallings, I Lenner, P Hallmans, G Mahlck, CG Wadell, G Schiller, J Dillner, J TI Type specificity and significance of different isotypes of serum antibodies to human papillomavirus capsids SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 17th International Papillomavirus Conference CY JAN 12, 1999 CL CHARLESTON, SOUTH CAROLINA ID CERVICAL INTRAEPITHELIAL NEOPLASIA; VIRUS-LIKE PARTICLES; INFECTION; CANCER; WOMEN; RISK; SEROREACTIVITY; RESPONSES; CARCINOMA; GAMMA AB Isotype-specific serum antibody responses against human papillomavirus (HPV) type 16 were evaluated by use of cross-sectional, prospective, and population-based seroepidemiologic studies. IgG1 and IgA were the most abundant isotypes. No sample contained IgG2, and <25 samples contained IgG3 or IgM. Total IgG, IgA, and IgG1 were HPV type specific and were associated with HPV-16 DNA (odds ratios [ORs], 5.4, 5.0, and 5.9, respectively; P<.001)but not with other HPV DNA (ORs, 1.2, 1.2, and 0.8, respectively; P value was not significant). Total IgG and IgG1 were strongly associated with number of lifetime sex partners (P <.001); IgA was only associated with number of recent sex partners and lifetime sex partners among younger women. Total IgG, IgG1, and IgA were associated with cervical intraepithelial neoplasia type III and also predicted risk of future cervical neoplasia. Ige and IgG1 appeared to mark lifetime cumulative exposure, whereas IgA may mark recent or ongoing infection. C1 Karolinska Inst, Ctr Microbiol & Tumor Biol, Lab Tumor Virus Epidemiol, S-17177 Stockholm, Sweden. Swedish Inst Infect Dis Control, Dept Bacteriol, Stockholm, Sweden. Univ Hosp No Sweden, Dept Virol, Umea, Sweden. Univ Hosp No Sweden, Dept Publ Hlth & Clin Med, Umea, Sweden. Univ Hosp No Sweden, Med Bank, Umea, Sweden. Natl Publ Hlth Inst, Helsinki, Finland. NCI, Cellular Oncol Lab, Bethesda, MD 20892 USA. Tampere Univ, FIN-33101 Tampere, Finland. RP Dillner, J (reprint author), Karolinska Inst, Ctr Microbiol & Tumor Biol, Lab Tumor Virus Epidemiol, Box 280, S-17177 Stockholm, Sweden. NR 31 TC 28 Z9 30 U1 0 U2 1 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD FEB PY 2000 VL 181 IS 2 BP 456 EP 462 DI 10.1086/315232 PG 7 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 292BT UT WOS:000085774800007 PM 10669326 ER PT J AU Stiehm, ER Fletcher, CV Mofenson, LM Palumbo, PE Kang, MH Fenton, T Sapan, CV Meyer, WA Shearer, WT Hawkins, E Fowler, MG Bouquin, P Purdue, L Sloand, EM Nemo, GJ Wara, D Bryson, YJ Starr, SE Petru, A Burchett, S AF Stiehm, ER Fletcher, CV Mofenson, LM Palumbo, PE Kang, MH Fenton, T Sapan, CV Meyer, WA Shearer, WT Hawkins, E Fowler, MG Bouquin, P Purdue, L Sloand, EM Nemo, GJ Wara, D Bryson, YJ Starr, SE Petru, A Burchett, S CA Pediat AIDS Trials Grp Protocol 273 TI Use of human immunodeficiency virus (HIV) human hyperimmune immunoglobulin in HIV type 1-infected children (Pediatric AIDS Clinical Trials Group Protocol 273) SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT Annual Meeting of the American-Pediatric-Society/Society-for-Pediatric-Research CY MAY-JUN -, 1999 CL SAN FRANCISCO, CALIFORNIA SP Amer Pediat Soc, Soc Pediat Res ID HUMAN MONOCLONAL-ANTIBODY; PASSIVE IMMUNOTHERAPY; PLASMA RICH; SERONEGATIVE PLASMA; INFECTION; PHARMACOKINETICS; NEUTRALIZATION; TRANSFUSIONS; IMMUNIZATION; PREVENTION AB The clinical, immunologic, and virologic effects and the pharmacokinetics of human immunodeficiency virus (HIV) human hyperimmune immunoglobulin (HIVIG) were assessed in 30 HIV-infected children aged 2-11 years. An had moderately advanced disease with an immune complex-dissociated (ICD) p24 antigen >70 pg/mL and were on stable antiviral therapy. Three groups of 10 children received 6 monthly infusions of 200, 400, or 800 mg/kg of HIVIG, and serial immunologic and virologic assays were performed. HIVIG doses as high as 800 mg/kg were safe and well tolerated. The half-life of HIVIG, determined by serial p24 antibody titers, was 13-16 days, the volume of distribution was 102-113 mL/kg, and clearance was 5.6-6.0 mL/kg/day. Plasma ICD p24 decreased during the infusions, but CD4 cell levels, plasma RNA copy number, cellular virus, immunoglobulin levels, and neutralizing antibody titers were minimally affected by the infusions. Clinical status did not change during the 3-month infusion and 3-month follow-up periods. C1 Univ Calif Los Angeles, Childrens Hosp, Los Angeles, CA 90095 USA. Univ Calif San Francisco, San Francisco, CA 94143 USA. Childrens Hosp, Oakland, CA 94609 USA. Univ Minnesota, Minneapolis, MN USA. NICHHD, Bethesda, MD 20892 USA. NIAID, Bethesda, MD 20892 USA. NHLBI, Bethesda, MD 20892 USA. Quest Diagnost Inc, Baltimore, MD USA. New Jersey Med Sch, Newark, NJ USA. Childrens Hosp, Boston, MA 02115 USA. N Amer Biol, Boca Raton, FL USA. Baylor Coll Med, Houston, TX 77030 USA. Childrens Hosp, Philadelphia, PA 19104 USA. RP Stiehm, ER (reprint author), Univ Calif Los Angeles, Childrens Hosp, 22-387 MDCC,10833 Le Conte Ave, Los Angeles, CA 90095 USA. OI Mofenson, Lynne/0000-0002-2818-9808 FU NCRR NIH HHS [RR-00865] NR 25 TC 18 Z9 18 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD FEB PY 2000 VL 181 IS 2 BP 548 EP 554 DI 10.1086/315224 PG 7 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 292BT UT WOS:000085774800019 PM 10669338 ER PT J AU King, JC Treanor, J Fast, PE Wolff, M Yan, LH Iacuzio, D Readmond, B O'Brien, D Mallon, K Highsmith, WE Lambert, JS Belshe, RB AF King, JC Treanor, J Fast, PE Wolff, M Yan, LH Iacuzio, D Readmond, B O'Brien, D Mallon, K Highsmith, WE Lambert, JS Belshe, RB TI Comparison of the safety, vaccine virus shedding, and immunogenicity of influenza virus vaccine, trivalent, types A and B, live cold-adapted, administered to human immunodeficiency virus (HIV)-infected and non-HIV-infected adults SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID DOUBLE-BLIND; VIRAL LOAD; CHILDREN AB Fifty-seven human immunodeficiency virus (HIV)-infected (CDC class A1-2) and 54 non-HIV-infected adults, not prescreened for influenza susceptibility, were randomized to receive trivalent live attenuated influenza vaccine (LAIV) or placebo intranasally, LAIV was safe and well tolerated with no serious adverse events attributable to vaccine, Reactogenicity rates were similar in LAIV and placebo recipients except that runny nose/nasal congestion was significantly more common in LAIV recipients regardless of HIV status. No prolonged shedding of LAIV was observed in HIV-infected participants. HIV RNA levels were not increased and CD4 counts were not decreased in HIV-infected LAIV recipients compared with placebo recipients after immunization. Shedding of LAIV and increases in antibody titers were infrequent, consistent with prior experience in unscreened adults. The data suggest that inadvertent vaccination with LAIV in relatively asymptomatic HIV-infected adults would not be associated with frequent significant adverse events. C1 Univ Maryland, Med Ctr, Sch Med, Ctr Vaccine Dev, Baltimore, MD 21201 USA. EMMES Corp, Potomac, MD USA. NIAID, NIH, Bethesda, MD 20892 USA. Univ Rochester, Med Ctr, Rochester, NY 14642 USA. Aviron, Mt View, CA USA. St Louis Univ, Sch Med, St Louis, MO USA. RP King, JC (reprint author), Univ Maryland, Med Ctr, Sch Med, Ctr Vaccine Dev, 700 W Lombard St, Baltimore, MD 21201 USA. RI Highsmith, William/B-6175-2008 FU NIAID NIH HHS [AI-45248, AI-45250, AI-45251] NR 15 TC 75 Z9 80 U1 0 U2 1 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD FEB PY 2000 VL 181 IS 2 BP 725 EP 728 DI 10.1086/315246 PG 4 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 292BT UT WOS:000085774800044 PM 10669363 ER PT J AU Wanishsawad, C Zhou, YF Epstein, SE AF Wanishsawad, C Zhou, YF Epstein, SE TI Chlamydia pneumoniae-induced transactivation of the major immediate early promoter of cytomegalovirus: Potential synergy of infectious agents in the pathogenesis of atherosclerosis SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID SMOOTH-MUSCLE CELLS; LOW-DENSITY-LIPOPROTEIN; ENDOTHELIAL-CELLS; EXPRESSION AB Both cytomegalovirus (CMV) and Chlamydia pneumoniae have been associated with atherosclerosis. CMV is believed to exist in host tissues in a latent state with periodic reactivation. This study was designed to determine whether C. pneumoniae infection stimulates the expression of CMV genes. Transactivation of the CMV major immediate early promoter (MIEP) is essential for viral gene expression and viral replication. HeLa cells were transfected with a construct containing a reporter gene (chloramphenicol acetyl transferase) controlled by the MIEP. The cells were then infected with Chlamydia at 10(2)-10(6) infection-forming units (IFU) per well at various times before assay of MIEP activity (72 h after transfection). Peak transactivation occurred 6 h after infection at 10(4) IFU, C, pneumoniae increased MIEP activity in a dose-response manner; maximal increase was >2-fold. These results suggest that if CMV and C. pneumoniae do indeed contribute to atherosclerosis, their copresence may synergistically contribute to it. C1 NHLBI, Cardiol Branch, NIH, Bethesda, MD 20892 USA. RP Epstein, SE (reprint author), Washington Hosp Ctr, Inst Cardiovasc Res, 110 Irving St NW,Ste 4B-1, Washington, DC 20010 USA. NR 15 TC 7 Z9 11 U1 0 U2 2 PU UNIV CHICAGO PRESS PI CHICAGO PA 5720 SOUTH WOODLAWN AVE, CHICAGO, IL 60637-1603 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD FEB PY 2000 VL 181 IS 2 BP 787 EP 790 DI 10.1086/315235 PG 4 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 292BT UT WOS:000085774800059 PM 10669378 ER PT J AU Huang, CK Zhan, LJ Hannigan, MO Ai, YX Leto, TL AF Huang, CK Zhan, LJ Hannigan, MO Ai, YX Leto, TL TI P47(phox)-deficient NADPH oxidase defect in neutrophils of diabetic mouse strains, C57BL6/J-m db/db and db/+ SO JOURNAL OF LEUKOCYTE BIOLOGY LA English DT Article DE superoxide production; mutation ID CHRONIC GRANULOMATOUS-DISEASE; SH3 DOMAIN; MAP KINASE; P47(PHOX); LEPTIN; ACTIVATION; EXPRESSION; MUTATIONS; MODEL AB Deficiencies in neutrophil NADPH oxidase proteins have been demonstrated in humans with chronic granulomatous disease. However, no spontaneous mutation in murine NADPH oxidase has been reported. In this study we report that neutrophils from the diabetic mouse strains, C57BL/6J-m heterozygous lean (lepr(db/+)) and homozygous obese (lepr(db/db)) mice produced no superoxide on stimulation. An absence of intact p47(phox) but not other oxidase proteins was ol,served in both mouse strains through the use of inmunoblotting. Molecular analysis by reverse transcriptase-polymerase chain reaction identified three abnormal p47 phox mRNA transcripts. Sequencing of genomic DNA of p47(phox) revealed a point mutation at the -2 position of exon 8, which is consistent with aberrant splicing of the p47(phox) transcript. These results indicate that the C57BL/6J-m db/db and db/+ mice are the first spontaneously derived murine model of NADPH oxidase deficiency involving a p47(phox) mutation. C1 Univ Connecticut, Ctr Hlth, Dept Pathol, Farmington, CT 06030 USA. NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. RP Huang, CK (reprint author), Univ Connecticut, Ctr Hlth, Dept Pathol, 263 Farmington Ave, Farmington, CT 06030 USA. FU NIAID NIH HHS [AI20943] NR 28 TC 67 Z9 67 U1 0 U2 1 PU FEDERATION AMER SOC EXP BIOL PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0741-5400 J9 J LEUKOCYTE BIOL JI J. Leukoc. Biol. PD FEB PY 2000 VL 67 IS 2 BP 210 EP 215 PG 6 WC Cell Biology; Hematology; Immunology SC Cell Biology; Hematology; Immunology GA 307AH UT WOS:000086630800010 PM 10670582 ER PT J AU Schultz, CJ Blanchette-Mackie, EJ Scow, RO AF Schultz, CJ Blanchette-Mackie, EJ Scow, RO TI Adrenal and liver in normal and cld/cld mice synthesize and secrete hepatic lipase, but the lipase is inactive in cld/cld mice SO JOURNAL OF LIPID RESEARCH LA English DT Article DE lipoprotein lipase; combined lipase deficiency; immunofluorescence microscopy; newborn mice ID HEPARIN-RELEASABLE LIPASE; CULTURED BROWN ADIPOCYTES; HORMONE-SENSITIVE LIPASE; LIPOPROTEIN-LIPASE; RAT-LIVER; TRIGLYCERIDE LIPASE; TRIACYLGLYCEROL LIPASE; DEFICIENCY CLD; MESSENGER-RNA; NEWBORN MICE AB Combined lipase deficiency (cld) is a recessive mutation in mice that causes a severe lack of lipoprotein lipase (LPL) and hepatic lipase (HL) activities, hyperlipemia, and death within 3 days after birth. Earlier studies showed that inactive LPL and HL were synthesized by cld/ cld tissues and that LPL synthesized by clcl/cld brown adipocytes was retained in their ER, We report here a study of HL in liver, adrenal, and plasma of normal newborn and cld/cld mice, Immunofluorescence studies showed HL was present in extracellular space, but not in cells, in liver and adrenal of both normal and cld/cld mice. When protein secretion was blocked with monensin, HL was retained intracellularly in liver cell cultures and in incubated adrenal tissues of both groups of mice. These findings demonstrated that HL was synthesized and secreted by liver and adrenal cells in normal newborn and cld/cld mice. HL activities in liver, adrenal, and plasma in cld/cld mice were very low, < 8% of that in normal newborn mice, indicating that HL synthesized and secreted by cld/cld cells was inactive. Livers of both normal newborn and cld/cld mice synthesized LPL, but the level of LPL activity in cld/cld liver was very low < 9% of that in normal liver. Immunofluorescence studies showed that LPL was present intracellularly in liver of cld/cld mice, indicating that LPL was synthesized but not secreted by cld/ cld liver cells. Immunofluorescent LPL was not found in normal newborn liver cells unless the cells were treated with monensin, thus demonstrating that normal Liver cells synthesized and secreted LPL, Livers of both groups of mice contained an unidentified alkaline lipase activity which accounted for 34-54% of alkaline lipase activity in normal and 65% of that in cld/cld livers. Our findings indicate that liver and adrenal cells synthesized and secreted HL in both normal newborn and cld/cld mice, but the lipase was inactive in cld/cld mice. That cld/cld liver cells secreted inactive HL while retaining inactive LPL indicates that these closely related lipases were processed differently. C1 NIDDKD, Cellular & Dev Biol Lab, NIH, Bethesda, MD 20892 USA. NIDDKD, Cell Biochem Lab, NIH, Bethesda, MD 20892 USA. RP Schultz, CJ (reprint author), NIDDKD, Cellular & Dev Biol Lab, NIH, Bethesda, MD 20892 USA. NR 67 TC 10 Z9 10 U1 0 U2 0 PU LIPID RESEARCH INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0022-2275 J9 J LIPID RES JI J. Lipid Res. PD FEB PY 2000 VL 41 IS 2 BP 214 EP 225 PG 12 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 287MR UT WOS:000085510200007 PM 10681405 ER PT J AU Tjandra, N Marquardt, J Clore, GM AF Tjandra, N Marquardt, J Clore, GM TI Direct refinement against proton-proton dipolar couplings in NMR structure determination of macromolecules SO JOURNAL OF MAGNETIC RESONANCE LA English DT Article ID ORIENTED MACROMOLECULES; METHYLENE PROTONS; RESTRAINTS; MAGNITUDE; DISTANCES; PROTEINS AB The computational tools necessary for making use of H-1-H-1 dipolar couplings in macromolecular structure refinement are presented. Potentials are described for direct refinement against H-1-H-1 dipolar couplings of known sign as well as of unknown sign. In addition, a multiple potential is developed for prochiral protons whose stereospecific assignments are unknown. The utility of direct H-1-H-1 dipolar coupling refinement is illustrated using the small protein ubiquitin. It is shown that direct H-1-H-1 dipolar coupling refinement leads to improvements in the precision, accuracy, and quality of the resulting structures. C1 NHLBI, Biophys Chem Lab, NIH, Bethesda, MD 20892 USA. NIDDKD, Phys Chem Lab, NIH, Bethesda, MD 20892 USA. RP Tjandra, N (reprint author), NHLBI, Biophys Chem Lab, NIH, Bldg 3, Bethesda, MD 20892 USA. RI Clore, G. Marius/A-3511-2008 OI Clore, G. Marius/0000-0003-3809-1027 NR 23 TC 55 Z9 56 U1 0 U2 4 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1090-7807 J9 J MAGN RESON JI J. Magn. Reson. PD FEB PY 2000 VL 142 IS 2 BP 393 EP 396 DI 10.1006/jmre.1999.1985 PG 4 WC Biochemical Research Methods; Physics, Atomic, Molecular & Chemical; Spectroscopy SC Biochemistry & Molecular Biology; Physics; Spectroscopy GA 281MA UT WOS:000085163300026 PM 10648162 ER PT J AU Gribar, JJ Ramachandra, M Hrycyna, CA Dey, S Ambudkar, SV AF Gribar, JJ Ramachandra, M Hrycyna, CA Dey, S Ambudkar, SV TI Functional characterization of glycosylation-deficient human P-glycoprotein using a vaccinia virus expression system SO JOURNAL OF MEMBRANE BIOLOGY LA English DT Article DE multidrug resistance; P-glycoprotein; drug transport; glycosylation; vaccinia virus ID MULTIDRUG-RESISTANCE; N-GLYCOSYLATION; INSECT CELLS; HUMAN MDR1; TRANSPORTER; MUTANTS; ASSOCIATION; PROTEASOME; STABILITY; CALNEXIN AB P-glycoprotein (P-gp), the product of human MDR1 gene, which functions as an ATP-dependent drug efflux pump, is N-linked glycosylated at asparagine residues 91, 94, and 99 located within the first extracellular loop. We report here the biochemical characterization of glycosylation-deficient (Gly(-)) P-gp using a vaccinia virus based transient expression system. The staining of HeLa cells expressing Gly(-) P-gp (91, 94, and 99N-->Q), with P-gp specific monoclonal antibodies, MRK-16, UIC2 and 4E3 revealed a 40 to 50% lower cell-surface expression of mutant P-gp compared to the wild-type protein. The transport function of Gly(-) P-gp, assessed using a variety of fluorescent compounds indicated that the substrate specificity of the pump was not affected by the lack of glycosylation. Additional mutants, Gly(-) D (91, 94, 99N-->D) and Gly(-) Delta (91, 94, 99 N deleted) were generated to verify that the reduced cell surface expression, as well as total expression, were not a result of the glutamine substitutions. Gly(-) D and Gly(-) Delta Pgps were also expressed to the same level as the Gly(-) mutant protein. S-35-Methionine/cysteine pulse-chase studies revealed a reduced incorporation of S-35-methionine/cysteine in full length Gly- P-gp compared to wild-type protein, but the half-life (similar to 3 hr) of mutant P-gp was essentially unaltered. Since treatment with proteasome inhibitors (MG-132, lactacystin) increased only the intracellular level of nascent, mutant P-gp, the decreased incorporation of S-35-methionine/cysteine in Gly(-) P-gp appears to be due to degradation of improperly folded mutant protein by the proteasome and endoplasmic reticulum-associated proteases, These results demonstrate that the unglycosylated protein, although expressed at lower levels at the cell surface, is functional and suitable for structural studies. C1 NCI, Cell Biol Lab, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Ambudkar, SV (reprint author), NCI, Cell Biol Lab, Div Basic Sci, NIH, Bldg 37, Bethesda, MD 20892 USA. RI Ambudkar, Suresh/B-5964-2008 NR 36 TC 70 Z9 71 U1 0 U2 5 PU SPRINGER VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0022-2631 J9 J MEMBRANE BIOL JI J. Membr. Biol. PD FEB 1 PY 2000 VL 173 IS 3 BP 203 EP 214 DI 10.1007/s002320001020 PG 12 WC Biochemistry & Molecular Biology; Cell Biology; Physiology SC Biochemistry & Molecular Biology; Cell Biology; Physiology GA 281KF UT WOS:000085158100004 PM 10667916 ER PT J AU Sobel, ME AF Sobel, ME TI New challenges for biomedical research utilizing human biological materials SO JOURNAL OF MOLECULAR DIAGNOSTICS LA English DT Article ID TISSUE C1 NCI, Pathol Lab, NIH, Mol Pathol Sect, Bethesda, MD 20892 USA. RP Sobel, ME (reprint author), NCI, Pathol Lab, NIH, Mol Pathol Sect, Bldg 10,Rm 2A33,10 Ctr Dr MSC 1500, Bethesda, MD 20892 USA. NR 9 TC 5 Z9 5 U1 0 U2 4 PU AMER SOC INVESTIGATIVE PATHOLOGY, INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3993 USA SN 1525-1578 J9 J MOL DIAGN JI J. Mol. Diagn. PD FEB PY 2000 VL 2 IS 1 BP 1 EP 4 DI 10.1016/S1525-1578(10)60608-3 PG 4 WC Pathology SC Pathology GA 365EG UT WOS:000089935000001 PM 11272896 ER PT J AU Fu, WM Killen, M Culmsee, C Dhar, S Pandita, TK Mattson, MP AF Fu, WM Killen, M Culmsee, C Dhar, S Pandita, TK Mattson, MP TI The catalytic subunit of telomerase is expressed in developing brain neurons and serves a cell survival-promoting function SO JOURNAL OF MOLECULAR NEUROSCIENCE LA English DT Article DE Alzheimer's disease; apoptosis; caspase; cerebral cortex; hippocampus; mitochondrial transmembrane potential; neurotrophic factor; Parkinson's disease ID PROSTATE APOPTOSIS RESPONSE-4; HIPPOCAMPAL-NEURONS; LIFE-SPAN; DEVELOPMENTAL REGULATION; PARKINSONS-DISEASE; IMMORTAL CELLS; DEATH; DIFFERENTIATION; P53; DEGENERATION AB Telomerase, a specialized reverse transcriptase (RT) linked to cell immortalization and cancer, has been thought not to be expressed in postmitotic cells. We now report that telomerase activity and its essential catalytic subunit, telomerase reverse transcriptase (TERT), are expressed in neurons in the brains of rodents during embryonic and early postnatal development, and are subsequently downregulated. Suppression of TERT expression in cultured embryonic hippocampal neurons increases their vulnerability to apoptosis and excitotoxicity, Overexpression of TERT in PC12 cells suppresses apoptosis induced by trophic factor withdrawal. TERT exerts its anti-apoptotic action at an early stage of the cell death process prior to mitochondrial dysfunction and caspase activation. TERT may serve a neuron survival-promoting function in the developing brain, and downregulation of TERT in the adult brain may contribute to increased neuronal vulnerability in various age-related neurodegenerative disorders. C1 Univ Kentucky, Sanders Brown Res Ctr Aging, Lexington, KY 40536 USA. Columbia Univ Coll Phys & Surg, Ctr Radiol Res, New York, NY 10032 USA. NIA, Neurosci Lab, Baltimore, MD 21224 USA. RP Mattson, MP (reprint author), Univ Kentucky, Sanders Brown Res Ctr Aging, Lexington, KY 40536 USA. RI Mattson, Mark/F-6038-2012 NR 43 TC 114 Z9 121 U1 0 U2 8 PU HUMANA PRESS INC PI TOTOWA PA 999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USA SN 0895-8696 J9 J MOL NEUROSCI JI J. Mol. Neurosci. PD FEB-APR PY 2000 VL 14 IS 1-2 BP 3 EP 15 DI 10.1385/JMN:14:1-2:003 PG 13 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 319EM UT WOS:000087328600002 PM 10854032 ER PT J AU Gozes, I Brenneman, DE AF Gozes, I Brenneman, DE TI A new concept in the pharmacology of neuroprotection SO JOURNAL OF MOLECULAR NEUROSCIENCE LA English DT Article DE neuroprotection; femtomolar; activity-dependent; VIP; ADNF ID VASOACTIVE-INTESTINAL-PEPTIDE; DEPENDENT NEUROTROPHIC FACTOR; CYCLASE-ACTIVATING POLYPEPTIDE; CORTICAL-NEURONS; MESSENGER-RNA; FACTOR ADNF; CELL-DEATH; EXPRESSION; ASTROCYTES; RECEPTOR AB Vasoactive intestinal peptide (VIP), originally discovered in the intestine as a peptide of 28 amino acids, was later found to be a major brain peptide having neuroprotective activities. To exert neuroprotective activity, VIP requires glial cells secreting neuroprotective proteins. Activity-dependent: neurotrophic factor (ADNF) is a recently isolated factor secreted by glial cells under the action of VIP. This protein, isolated by sequential chromatographic methods, was named activity-dependent neurotrophic factor since it protected neurons from death associated with blockade of electrical activity. A fourteen-amino-acid fragment of ADNF (ADNF-14) and the more potent, nine-amino-acid derivative (ADNF-9), exhibit activity that surpasses that of the parent protein with regard to potency and a broader range of effective concentration. Furthermore, the peptides exhibit protective activity in Alzheimer's disease-related systems (e.g., beta-amyloid toxicity and apoplipoprotein E deficiencies, genes that have been associated with Alzheimer's disease onset and progression). ADNP is another glial mediator of VIP-associated neuroprotection. NAP, an eight-amino-acid peptide derived from ADNP (sharing structural and functional similarities with ADNF-9), was identified as the most potent neuroprotectant described to-date in an animal model of apolipoprotein E-deficiency (knock-out mice). These femtomolar-acting peptides form a basis for a new concept in pharmacology: femtomolar neuroprotection. C1 Tel Aviv Univ, Sackler Sch Med, Dept Clin Biochem, IL-69978 Tel Aviv, Israel. NICHHD, Dev Neurobiol Lab, Sect Dev & Mol Pharmcol, NIH, Bethesda, MD 20892 USA. RP Gozes, I (reprint author), Tel Aviv Univ, Sackler Sch Med, Dept Clin Biochem, IL-69978 Tel Aviv, Israel. NR 37 TC 86 Z9 91 U1 3 U2 5 PU HUMANA PRESS INC PI TOTOWA PA 999 RIVERVIEW DRIVE SUITE 208, TOTOWA, NJ 07512 USA SN 0895-8696 J9 J MOL NEUROSCI JI J. Mol. Neurosci. PD FEB-APR PY 2000 VL 14 IS 1-2 BP 61 EP 68 DI 10.1385/JMN:14:1-2:061 PG 8 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 319EM UT WOS:000087328600007 PM 10854037 ER PT J AU Moncman, CL Wang, K AF Moncman, CL Wang, K TI Architecture of the thin filament-Z-line junction: lessons from nebulette and nebulin homologies SO JOURNAL OF MUSCLE RESEARCH AND CELL MOTILITY LA English DT Article ID SKELETAL-MUSCLE SARCOMERE; MOLECULAR-WEIGHT PROTEINS; PURIFIED ALPHA-ACTININ; VERTEBRATE Z-DISCS; C-TERMINAL REGION; STRIATED-MUSCLE; 3-DIMENSIONAL STRUCTURE; INVIVO PHOSPHORYLATION; BINDING-PROPERTIES; CARDIAC-MUSCLE AB Nebulette and nebulin are homologous proteins associated with the Z-lines of cardiac and skeletal muscle sarcomeres. Although these proteins share similar to 70% sequence homology and an identical domain layout, nebulette is one-tenth the size of nebulin. To define structurally important features of these proteins in terms of the Z-line architecture, we have analyzed the primary structure of nebulette and nebulin from a variety of species and developmental stages. Alignment of the 35 residue nebulin-like modules from both proteins demonstrates that the individual modules share 30-90% homology across the six proteins analyzed. In addition, this analysis demonstrates a number of areas in which the identity across the six proteins is as high as 75%. These areas may be important signals for Z-line assembly and function in the striated muscles. Significantly, most of the areas of high identity also coincide with consensus phosphorylation sites. To evaluate if nebulette, like nebulin, exhibits tissue-specific and developmental specific polymorphism, a series of immunoblot assays were performed. These data demonstrate that nebulettes from different portions of the heart are the same size. Comparison of nebulette from embryonic and adult cardiac muscle also demonstrates that this protein does not appear to vary in size with developmental stage. Consistent with the large number of consensus phosphorylation sites identified in the nebulette primary structure, we find that nebulette is phosphorylated in the cardiac muscle at serine and threonine residues. These data and sequence analyses are discussed in terms of current models for Z-line architecture. C1 Univ Texas, Dept Chem & Biochem, Austin, TX 78712 USA. RP NIAMS, NIH, Bldg 6 Rm 401, Bethesda, MD 20892 USA. EM wangk@exchange.nih.gov FU NIAMS NIH HHS [AR43514] NR 59 TC 19 Z9 19 U1 0 U2 1 PU SPRINGER PI DORDRECHT PA VAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDS SN 0142-4319 EI 1573-2657 J9 J MUSCLE RES CELL M JI J. Muscle Res. Cell Motil. PD FEB PY 2000 VL 21 IS 2 BP 153 EP 169 DI 10.1023/A:1005697226465 PG 17 WC Cell Biology SC Cell Biology GA 324AZ UT WOS:000087600000005 PM 10961839 ER PT J AU Gustafson, KR Walton, LK Sowder, RC Johnson, DG Pannell, LK Cardellina, JH Boyd, MR AF Gustafson, KR Walton, LK Sowder, RC Johnson, DG Pannell, LK Cardellina, JH Boyd, MR TI New circulin macrocyclic polypeptides from Chassalia parvifolia SO JOURNAL OF NATURAL PRODUCTS LA English DT Article AB Four new macrocyclic polypeptides were isolated and identified from an extract of the tropical tree Chassalia parvifolia. Circulins C-F are 29-30 amino acid cyclic peptides in which the entire primary amino acid chain is covalently cyclized via peptide bonds. Their structures were deduced from a combination of FABMS analyses, N-terminal Edman degradation, endoproteinase digestion, and amino acid analyses. All the peptides share a high degree of sequence homology and contain six cysteine residues forming three intramolecular disulfide bridges. Circulins C-F inhibited the cytopathic effects of in vitro HIV-1 infection with EC50 values of 50-275 nM. C1 NCI, Frederick Canc Res & Dev Ctr, Lab Drug Discovery Res & Dev, Dev Therapeut Program,Div Canc Treatement & Diagn, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, Aids Vaccine Program, SAIC Inc, Frederick, MD 21702 USA. NIDDK, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. RP Boyd, MR (reprint author), NCI, Frederick Canc Res & Dev Ctr, Lab Drug Discovery Res & Dev, Dev Therapeut Program,Div Canc Treatement & Diagn, Bldg 1052,Room 102, Frederick, MD 21702 USA. FU NCI NIH HHS [N01-CO-56000] NR 17 TC 81 Z9 89 U1 0 U2 8 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0163-3864 J9 J NAT PROD JI J. Nat. Prod. PD FEB PY 2000 VL 63 IS 2 BP 176 EP 178 DI 10.1021/np990432r PG 3 WC Plant Sciences; Chemistry, Medicinal; Pharmacology & Pharmacy SC Plant Sciences; Pharmacology & Pharmacy GA 287KW UT WOS:000085505900002 PM 10691702 ER PT J AU Mattson, MP Culmsee, C Yu, ZF Camandola, S AF Mattson, MP Culmsee, C Yu, ZF Camandola, S TI Roles of nuclear factor kappa B in neuronal survival and plasticity SO JOURNAL OF NEUROCHEMISTRY LA English DT Review DE apoptosis; excitotoxicity; hippocampus; long-term potentiation; neurotrophic factor; oxidative stress; transcription factor; tumor necrosis factor ID TUMOR-NECROSIS-FACTOR; AMYLOID-PRECURSOR PROTEIN; MANGANESE SUPEROXIDE-DISMUTASE; DNA-BINDING ACTIVITY; INDUCED CELL-DEATH; BETA-PEPTIDE TOXICITY; ISCHEMIC BRAIN INJURY; NERVE GROWTH-FACTOR; INDUCED APOPTOSIS; HIPPOCAMPAL-NEURONS AB The transcription factor nuclear factor kappa B (NF-kappa B) is moving to the forefront of the fields of apoptosis and neuronal plasticity because of recent findings showing that activation of NF-kappa B prevents neuronal apoptosis in various cell culture and in vivo models and because NF-kappa B is activated in association with synaptic plasticity. Activation of NF-kappa B was first shown to mediate antiapoptotic actions of tumor necrosis factor in cultured neurons and was subsequently shown to prevent death of various nonneuronal cells. NF-kappa B is activated by several cytokines and neurotrophic factors and in response to various cell stressors, Oxidative stress and elevation of intracellular Calcium levels are particularly important inducers of NF-kappa B activation, Activation of NF-kappa B can interrupt apoptotic biochemical cascades at relatively early steps, before mitochondrial dysfunction acid oxyradical production; Gene targets for NF-kappa B that may mediate its antiapoptotic actions include the antioxidant enzyme manganese superoxide dismutase, members of the inhibitor of apoptosis family of proteins, and the calcium-binding protein calbindin D28k. NF-kappa B is activated by synaptic activity and may play important roles in the process of learning and memory. The available data identify NF-kappa B as an important regulator of evolutionarily conserved biochemical and molecular cascades designed to prevent cell death and promote neuronal plasticity. Because NF-kappa B may play roles in a range of neurological disorders that involve neuronal degeneration and/or perturbed synaptic function, pharmacological and genetic manipulations of NF-kappa B signaling are being developed that may prove valuable in treating disorders ranging from Alzheimer's disease to schizophrenia. C1 Univ Kentucky, Sanders Brown Res Ctr Aging, Lexington, KY USA. Univ Kentucky, Dept Anat & Neurobiol, Lexington, KY USA. RP Mattson, MP (reprint author), NIA, Neurosci Lab, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012 NR 125 TC 360 Z9 385 U1 4 U2 12 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD FEB PY 2000 VL 74 IS 2 BP 443 EP 456 DI 10.1046/j.1471-4159.2000.740443.x PG 14 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 274KU UT WOS:000084765700001 PM 10646495 ER PT J AU Nakai, M Qin, ZH Chen, JF Wang, YM Chase, TN AF Nakai, M Qin, ZH Chen, JF Wang, YM Chase, TN TI Kainic acid-induced apoptosis in rat striatum is associated with nuclear factor-kappa B activation SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE kainic acid; apoptosis; neurodegeneration; nuclear transcription factor-kappa B (NF-kappa B); I kappa B-alpha; NF-kappa B p65; c-Myc; p53 ID METHYL-D-ASPARTATE; CEREBELLAR GRANULE NEURONS; CEREBRAL-ARTERY OCCLUSION; MESSENGER-RNA EXPRESSION; BETA-AMYLOID PRECURSOR; CORTICAL CELL-CULTURES; DNA FRAGMENTATION; TUMOR-SUPPRESSOR; QUINOLINIC ACID; HUNTINGTONS-DISEASE AB The present study evaluated whether nuclear factor-kappa B (NF-kappa B) activation contributes to the apoptotic-like death of striatal neurons induced by kainic acid (KA) receptor stimulation. Intrastriatally infused KA (1.25-5.0 nmol) produced substantial neuronal loss as indicated by an 8-73% decrease in 67-kDa glutamic acid decarboxylase (p < 0.05), KA (1.25-5.0 nmol) elicited internucleosomal DNA fragmentation that was inhibited by the AMPA/KA receptor antagonist NBQX (1,2,3,4-tetrahydro-6-nitro-2,3-dibenzo[f]quinoxaline-7-sulfonamide) but not by the NMDA receptor antagonist MK-801. A decrease in I kappa B-alpha. protein levels, which was accompanied by an increase in NF-kappa B binding activity, was found from 6 to 72 h after KA (2.5 nmol) infusion. NF-kappa B was composed mainly of p65 and c-Rel as revealed by supershift assay. In addition, c-Myc and p53 increased from five- to sevenfold from 24 to 72 h after KA (2.5 nmol) administration. Immunohistochemistry revealed high levels of c-Myc and p53 immunoreactivity, mainly in medium-sized striatal neurons. Pretreatment with the cell-permeable recombinant peptide NF-kappa B SN50 (5-20 mu g) blocked NF-kappa B nuclear translocation, but had no effect on AP-I binding. NF-kappa B SN50 also inhibited the KA-induced up-regulation of c-Myc and p53, as well as internucleosomal DNA fragmentation. The apoptotic-like destruction of rat striatal neurons induced by KA receptor stimulation thus appears to involve biochemical mechanisms similar to those mediating: the excitotoxic response to NMDA receptor stimulation. The present results provide additional support for the view that NF-kappa B activation contributes to c-Myc and p53 induction and subsequent apoptosis in an excitotoxic model of Huntington's disease. C1 NINCDS, Expt Therapeut Branch, NIH, Bethesda, MD 20892 USA. Massachusetts Gen Hosp, Mol Neurobiol Lab, Boston, MA 02114 USA. Harvard Univ, Sch Med, Boston, MA 02115 USA. RP Chase, TN (reprint author), NINCDS, Expt Therapeut Branch, NIH, Bldg 10,Rm 5C103,10 Ctr Dr,MSC 1406, Bethesda, MD 20892 USA. NR 72 TC 91 Z9 96 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD FEB PY 2000 VL 74 IS 2 BP 647 EP 658 DI 10.1046/j.1471-4159.2000.740647.x PG 12 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 274KU UT WOS:000084765700022 PM 10646516 ER PT J AU Nye, EJ Bornstein, SR Grice, JE Tauchnitz, R Hockings, GI Strakosch, CR Jackson, RV Torpy, DJ AF Nye, EJ Bornstein, SR Grice, JE Tauchnitz, R Hockings, GI Strakosch, CR Jackson, RV Torpy, DJ TI Interactions between the stimulated hypothalamic-pituitary-adrenal axis and leptin in humans SO JOURNAL OF NEUROENDOCRINOLOGY LA English DT Article DE leptin; hydrocortisone; adrenocorticotropin; naloxone; hypoglycaemia ID CORTICOTROPIN-RELEASING HORMONE; PLASMA LEPTIN; CORTISOL RELEASE; DEXAMETHASONE; ADRENOCORTICOTROPIN; GLUCOCORTICOIDS; VASOPRESSIN; SECRETION; HYPERINSULINEMIA; INHIBITION AB Leptin, produced by adipocytes, has homeostatic effects on body fat mass through inhibition of appetite and stimulation of the sympathetic nervous system. Several studies have reported that high-dose exogenous glucocorticoids increase circulating leptin concentrations in humans. Conversely, leptin has inhibitory effects on the hypothalamic-pituitary-adrenal (HPA) axis, both at the hypothalamic and adrenal levels, We hypothesized that acute hypercortisolism, in the physiological range, may not alter leptin secretion, Four stimuli of the HPA axis were administered to eight healthy male volunteers in a placebo-controlled study, On separate afternoons, in a randomised order, fasting subjects received i.v. injections of saline, naloxone (125 mu g/kg); vasopressin (0.0143 IU/kg); naloxone and vasopressin in combination; or insulin (0.15 U/kg; a dose sufficient to induce hypoglycaemia), Plasma concentrations of adrenocorticotrophic hormone (ACTH), cortisol and leptin were measured before and for 120 min after the injection. The cortisol secretory response was greatest after insulin-hypoglycaemia, this response was significantly greater than that following naloxone, naloxone/vasopressin, or vasopressin alone, Despite the cortisol release, leptin concentrations were not increased after any stimulus, Insulin-hypoglycaemia was associated with a decrease in leptin concentration at 60 and 90 min, while naloxone did not alter leptin concentrations. However, basal leptin concentrations were positively correlated with integrated ACTH and cortisol responses to naloxone, but did not correlate with ACTH or cortisol responses to the other stimuli. Thus acute elevations of plasma cortisol, in the physiological range, do not appear to influence plasma leptin concentrations. The fall in plasma leptin concentration after insulin-induced hypoglycaemia may reflect catecholamine secretion after this stimulus. C1 Univ Queensland, Greenslopes Private Hosp, Dept Med, Neuroendocrine Res Unit, Brisbane, Qld 4120, Australia. NICHD, Dev Endocrinol Branch, NIH, Bethesda, MD USA. Univ Leipzig, Leipzig, Germany. RP Torpy, DJ (reprint author), Univ Queensland, Greenslopes Private Hosp, Dept Med, Neuroendocrine Res Unit, Brisbane, Qld 4120, Australia. RI Grice, Jeffrey/F-6257-2010; Strakosch, Christopher/F-9457-2013 OI Grice, Jeffrey/0000-0002-8862-4461; Strakosch, Christopher/0000-0002-2319-4069 NR 29 TC 19 Z9 19 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0953-8194 J9 J NEUROENDOCRINOL JI J. Neuroendocrinol. PD FEB PY 2000 VL 12 IS 2 BP 141 EP 145 DI 10.1046/j.1365-2826.2000.00431.x PG 5 WC Endocrinology & Metabolism; Neurosciences SC Endocrinology & Metabolism; Neurosciences & Neurology GA 292YL UT WOS:000085823300006 PM 10718909 ER PT J AU Peters, TR Bodine, DM McDonagh, KT Lovett-Racke, A McFarland, HF McFarlin, DE Nienhuis, AW Racke, MK AF Peters, TR Bodine, DM McDonagh, KT Lovett-Racke, A McFarland, HF McFarlin, DE Nienhuis, AW Racke, MK TI Retrovirus mediated gene transfer of the self antigen MBP into the bone marrow of mice alters resistance to experimental autoimmune encephalomyelitis SO JOURNAL OF NEUROIMMUNOLOGY LA English DT Article DE experimental autoimmune encephalomyelitis; gene transfer; tolerance; myelin basic protein ID MYELIN BASIC-PROTEIN; EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS; LIMITING DILUTION ASSAYS; T-CELLS; TOLERANCE INDUCTION; NEGATIVE SELECTION; THYMIC EXPRESSION; ADOPTIVE TRANSFER; C57BL/6 MICE; MOUSE AB Experimental autoimmune encephalomyelitis (EAE) is a prototypic model of organ specific autoimmunity. MHC class II restricted T-cells directed against myelin basic protein (MBP) have been shown to cause EAE in susceptible strains of mice. We have asked whether the introduction of a gene encoding an autoantigen (MBP) into the hematopoetic stem cells of mice would result in tolerance to that protein. We have introduced cDNA encoding the 21.5 kDa isoform of MBP into the hematopoetic stem cells of B10.PL (73NS), SJL, and B10 mice by retrovirus-mediated gene transfer. Our experiments show expression of proviral MBP in peripheral blood and thymus following transplantation of genetically modified stem cells. Such expression does not result in deletion of MBP-specific T cells or tolerance to MBP, nor does it alter susceptibility to MBP-induced EAE in susceptible strains B10.PL and Sn. However, retrovirus-mediated gene transfer resulted in resistant B10 mice developing mild EAE. This report demonstrates that autoreactive MBP-specific T cells can be selected in the presence of endogenous antigen or an MBP-encoding retrovirus, (C) 2000 Elsevier Science B.V. All rights reserved. C1 NHLBI, Clin Hematol Branch, NIH, Bethesda, MD 20892 USA. NIH, Hematopoesis Sect, Lab Gene Transfer, Natl Ctr Human Genome Res, Bethesda, MD 20892 USA. Univ Michigan, Dept Internal Med, Ann Arbor, MI 48109 USA. NINDS, Neuroimmunol Branch, NIH, Bethesda, MD 20892 USA. St Jude Childrens Res Hosp, Memphis, TN 38101 USA. Washington Univ, Sch Med, Dept Neurol, St Louis, MO 63110 USA. RP Racke, MK (reprint author), Univ Texas, SW Med Ctr, Dept Neurol, 5323 Harry Hines Blvd, Dallas, TX 75235 USA. RI Lovett-Racke, Amy/E-3545-2011 NR 57 TC 10 Z9 10 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 FEB 1 PY 2000 VL 103 IS 1 BP 51 EP 62 DI 10.1016/S0165-5728(99)00181-2 PG 12 WC Immunology; Neurosciences SC Immunology; Neurosciences & Neurology GA 277MM UT WOS:000084937400007 PM 10674989 ER PT J AU Recanzone, GH Wurtz, RH AF Recanzone, GH Wurtz, RH TI Effects of attention on MT and MST neuronal activity during pursuit initiation SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID TEMPORAL VISUAL AREA; POSTERIOR PARIETAL CORTEX; EYE-MOVEMENTS; MACAQUE MONKEY; EXPANSION CONTRACTION; FUNCTIONAL-PROPERTIES; SELECTIVE ATTENTION; NEURAL MECHANISMS; TARGET SELECTION; RECEPTIVE-FIELD AB The responses of neurons in monkey extrastriate areas MT (middle temporal) and MST (medial superior temporal), and the initial metrics of saccadic and pursuit eye movements, have: previously been shown to be better predicted by vector averaging or winner-take-all models depending on the stimulus conditions. To investigate the potential influences of attention on the neuronal activity, we measured the responses of single MT and MST neurons under identical stimulus conditions when one of two moving stimuli was the target for a pursuit eye movement. We found the greatest attentional modulation across neurons when two stimuli moved through the receptive field (RF) of the neuron and the stimulus motion was initiated at least 450 ms before reaching the center of the RF. These conditions were the same as those in which a winner-take-all model better predicted both the eye movements and the underlying neuronal activity. The modulation was almost always an increase of activity, and it was about equally frequent in MT and MST. A modulation of >50% was observed in similar to 41% of MT neurons and 27% of MST neurons. Responses to all directions of motion were modulated so that the direction tuning curves in the attended and unattended conditions were similar. Changes in the background activity with target selection were small and unlikely to account for the observed attentional modulation. In contrast, there was little change in the neuronal response with attention when the stimulus reached the RF center 150 ms after motion onset, which was also the condition in which the vector average model better predicted the initial eye movements and the activity of the neurons. These results are consistent with a competition model of attention in which top-down attention acts on the activity of one of two competing populations of neurons activated by the bottom-up input from peripheral stimuli. They suggest that there is-a minimal separation of the populations necessary before attention can act on one population, similar to that required to produce a winner-take-all mode of behavior in pursuit initiation. The present experiments also suggest that it takes several hundred milliseconds to develop this top-down attention effect. C1 NEI, Sensorimotor Res Lab, NIH, Bethesda, MD 20892 USA. Univ Calif Davis, Ctr Neurosci, Davis, CA 95616 USA. Univ Calif Davis, Sect Neurobiol Physiol & Behav, Davis, CA 95616 USA. RP Wurtz, RH (reprint author), NEI, Sensorimotor Res Lab, NIH, 9000 Rockville Pike,Bldg 49,Rm 2A50, Bethesda, MD 20892 USA. FU NIDCD NIH HHS [DC-02371] NR 44 TC 90 Z9 92 U1 1 U2 3 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3077 J9 J NEUROPHYSIOL JI J. Neurophysiol. PD FEB PY 2000 VL 83 IS 2 BP 777 EP 790 PG 14 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 285TJ UT WOS:000085403900014 PM 10669493 ER PT J AU Latham, PE Richmond, BJ Nelson, PG Nirenberg, S AF Latham, PE Richmond, BJ Nelson, PG Nirenberg, S TI Intrinsic dynamics in neuronal networks. I. Theory SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID SPIKE FREQUENCY ADAPTATION; RECEPTIVE-FIELD PROPERTIES; PRIMARY VISUAL-CORTEX; LOW FIRING RATES; NEOCORTICAL NEURONS; SYNAPTIC-INTERACTIONS; RESPIRATORY RHYTHM; CELL-CULTURE; CAT; INTERNEURONS AB Many networks in the mammalian nervous system remain active in the absence of stimuli. This activity falls into two main patterns: steady firing at low rates and rhythmic bursting. How are these firing patterns generated? Specifically, how do dynamic interactions between excitatory and inhibitory neurons produce these firing patterns, and how do networks switch from one firing pattern to the other? We investigated these questions theoretically by examining the intrinsic dynamics of large networks of neurons. Using both a semianalytic model based on mean firing rate dynamics and simulations with large neuronal networks, we found that the dynamics, and thus the firing patterns, are controlled largely by one parameter, the fraction of endogenously active cells. When no endogenously active cells are present, networks are either silent or fire at a high rate; as the number of endogenously active cells increases, there is a transition to bursting; and, with a further increase, there is a second transition to steady firing at a low rate. A secondary role is played by network connectivity, which determines whether activity occurs at a constant mean firing rate or oscillates around that mean. These conclusions require only conventional assumptions: excitatory input to a neuron increases its firing rate, inhibitory input decreases it, and neurons exhibit spike-frequency adaptation. These conclusions also lead to two experimentally testable predictions: 1) isolated networks that fire at low rates must contain endogenously active cells and 2) a reduction in the fraction of endogenously active cells in such networks must lead to bursting. C1 Univ Calif Los Angeles, Dept Neurobiol, Los Angeles, CA 90095 USA. NIMH, Neuropsychol Lab, NIH, Bethesda, MD 20892 USA. NICHHD, Dev Neurobiol Lab, NIH, Bethesda, MD 20892 USA. RP Latham, PE (reprint author), Univ Calif Los Angeles, Dept Neurobiol, Box 95-1763, Los Angeles, CA 90095 USA. NR 58 TC 176 Z9 176 U1 0 U2 4 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3077 J9 J NEUROPHYSIOL JI J. Neurophysiol. PD FEB PY 2000 VL 83 IS 2 BP 808 EP 827 PG 20 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 285TJ UT WOS:000085403900017 PM 10669496 ER PT J AU Latham, PE Richmond, BJ Nirenberg, S Nelson, PG AF Latham, PE Richmond, BJ Nirenberg, S Nelson, PG TI Intrinsic dynamics in neuronal networks. II. Experiment SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID CENTRAL-NERVOUS-SYSTEM; SPINAL-CORD NEURONS; SYNAPSE FORMATION; TETANUS TOXIN; RECEPTORS; CULTURES; CONNECTIONS; INVITRO AB Neurons in many regions of the mammalian CNS remain active in the absence of stimuli. This activity falls into two main patterns: steady firing at low rates and rhythmic bursting. How these firing patterns are maintained in the presence of powerful recurrent excitation, and how networks switch between them, is not well understood. In the previous paper, we addressed these issues theoretically; in this paper we address them experimentally. We found in both studies that a key parameter in controlling firing patterns is the fraction of endogenously active cells. The theoretical analysis indicated that steady firing rates are possible only when the fraction of endogenously active cells is above some threshold, that there is a transition to bursting when it falls below that threshold, and that networks becomes silent when the fraction drops to zero. Experimentally, we found that all steadily firing cultures contain endogenously active cells, and that reducing the fraction of such cells in steadily firing cultures causes a transition to bursting. The latter finding implies indirectly that the elimination of endogenously active cells would cause a permanent drop to zero firing rate. The experiments described here thus corroborate the theoretical analysis. C1 Univ Calif Los Angeles, Dept Neurobiol, Los Angeles, CA 90095 USA. NICHHD, Dev Neurobiol Lab, NIH, Bethesda, MD 20892 USA. NIMH, Neuropsychol Lab, NIH, Bethesda, MD 20892 USA. RP Latham, PE (reprint author), Univ Calif Los Angeles, Dept Neurobiol, Box 95-1763, Los Angeles, CA 90095 USA. NR 30 TC 67 Z9 67 U1 0 U2 1 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3077 J9 J NEUROPHYSIOL JI J. Neurophysiol. PD FEB PY 2000 VL 83 IS 2 BP 828 EP 835 PG 8 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 285TJ UT WOS:000085403900018 PM 10669497 ER PT J AU Gruart, A Schreurs, BG Del Toro, ED Delgado-Garcia, JM AF Gruart, A Schreurs, BG Del Toro, ED Delgado-Garcia, JM TI Kinetic and frequency-domain properties of reflex and conditioned eyelid responses in the rabbit SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID SLOW FINGER MOVEMENTS; NICTITATING-MEMBRANE; MOTOR OUTPUT; ALERT CAT; ORGANIZATION; MUSCLE; BLINKS; AFFERENTS; CORTEX; MEMORY AB Eyelid position and the electromyographic activity of the orbicularis oculi muscle were recorded unilaterally in rabbits during reflex and conditioned blinks. Air-puff-evoked blinks consisted of a fast downward phase followed sometimes by successive downward sags. The reopening phase had a much longer duration and slower peak velocity. Onset latency, maximum amplitude, peak velocity, and rise time of reflex blinks depended on the intensity and duration of the air puff-evoking stimulus. A flashlight focused on the eye also evoked reflex blinks, but not flashes of light, or tones. Both delayed and trace classical conditioning paradigms were used. For delayed conditioning, animals were presented with a 350-ms, 90-dB, 600-Hz tone, as conditioned stimulus (CS). For trace conditioning, animals were presented with a IO-ms, 1-k/cm(2) air puff, as CS. The unconditioned stimulus (US) consisted of a 100-ms, 3-k/cm(2) air puff. The stimulus interval between CS and US onsets was 250 ms. Conditioned responses (CRs) to tones were composed of downward sags that increased in number through the successive conditioning sessions. The onset latency of the CR decreased across conditioning at the same time as its maximum amplitude and its peak velocity increased, but the time-to-peak of the CR remained unaltered. The topography of CRs evoked by short, weak air puffs as the CS showed three different components: the alpha response to the CS, the CR, and the reflex response to the US. Through conditioning, CRs showed a decrease in onset latency, and an increase in maximum amplitude and peak velocity. The time-to-peak of the CR remained unchanged. A power spectrum analysis of reflex and conditioned blink acceleration profiles showed a significant approximate to 8-Hz oscillation within a broadband of frequencies between 4 and 15 Hz. Nose and mandible movements presented power spectrum profiles different from those characterizing reflex and conditioned blinks. It is concluded that eyelid reflex responses in the rabbit present significant differences from CRs in their profiles and metric properties, suggesting different neural origins, but that a common approximate to 8-Hz neural oscillator underlies lid motor performance. According to available data, the frequency of this putative oscillator seems to be related to the species size. C1 Univ Seville, Lab Neurociencia, Seville 41012, Spain. NINDS, Behav Neurosci Unit, NIH, Bethesda, MD 20892 USA. RP Gruart, A (reprint author), Fac Biol, Lab Neurociencia, Avda Reina Mercedes 6, Seville 41012, Spain. OI Schreurs, Bernard/0000-0002-5776-0807 NR 41 TC 64 Z9 65 U1 0 U2 2 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3077 J9 J NEUROPHYSIOL JI J. Neurophysiol. PD FEB PY 2000 VL 83 IS 2 BP 836 EP 852 PG 17 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 285TJ UT WOS:000085403900019 PM 10669498 ER PT J AU Dobi, A Palkovits, M Palkovits, CG Ring, MA Von Agoston, D AF Dobi, A Palkovits, M Palkovits, CG Ring, MA Von Agoston, D TI Septamer element-binding proteins in neuronal and glial differentiation SO JOURNAL OF NEUROSCIENCE LA English DT Article DE progenitors; neuron; glia; development; DNA molecular decoy; DNA bending ID CENTRAL-NERVOUS-SYSTEM; DOMAIN TRANSCRIPTION FACTORS; GENE-EXPRESSION; STEM-CELLS; PROENKEPHALIN GENE; DECOY STRATEGY; PURKINJE-CELLS; MESSENGER-RNA; CIS-ELEMENTS; DNA-BINDING AB Differentiation of progenitors into neurons and glia is regulated by interactions between regulatory DNA elements of neuron- and glia-specific genes and transcription factors that are differentially expressed by progenitors at progressive stages of neural development. We have identified a novel DNA regulatory element (TTTGCAT = septamer) present on the enkephalin (ENK), neuronal cell adhesion molecule, neurofilament of 68 kDa (NF68), growth-associated protein of 43 kDa, glial high-affinity glutamine transporter, tyrosine hydroxylase, etc., genes. When septamer function was blocked by introducing septamer competitor DNA into primary differentiating neural cultures, mRNA levels of ENK, NF68, and glial fibrillary acidic protein decreased by 50-80%, whereas no effect was seen using a control DNA. Septamer elements serve as binding sites for lineage-specific multimeric complexes assembled from three distinct nuclear proteins. Progenitors express a 16 kDa protein (p-sept) which binds to DNA as a homodimer (detected as the 32 kDa P-band). Cells that entered the neuronal lineage express an additional 29 kDa protein (n-sept) that binds to the homodimerized p-sept, and together they form a 62 kDa multimer (detected as N-band). Cells that entered the glial lineage express a distinct 23 kDa protein (g-sept), which along with the homodimerized p-sept form a 56 kDa multimer (observed as G-band). The binding of the distinct protein complexes (P, G, and N) to the septamer site causes a lineage-specific DNA bending (P = 53 degrees; G = 72 degrees; and N = 90 degrees), which may contribute to the regulatory effect of the septamer interaction. In summary, septamer and its binding proteins represent novel protein-DNA interactions that may contribute to the regulation of neuroglial differentiation in the developing mammalian CNS. C1 Uniformed Serv Univ Hlth Sci, Sch Med, Dept Anat & Cell Biol, Bethesda, MD 20814 USA. Uniformed Serv Univ Hlth Sci, Sch Med, Neurosci Program, Bethesda, MD 20814 USA. NICHHD, Dev Neurobiol Lab, NIH, Bethesda, MD 20892 USA. NIMH, Genet Lab, Bethesda, MD 20892 USA. RP Uniformed Serv Univ Hlth Sci, Sch Med, Dept Anat & Cell Biol, B2036, Bethesda, MD 20814 USA. EM vagoston@helix.nih.gov RI Palkovits, Miklos/F-2707-2013; OI Palkovits, Miklos/0000-0003-0578-0387 FU NICHD NIH HHS [Z01 HD 00712-03] NR 60 TC 11 Z9 11 U1 0 U2 1 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD FEB 1 PY 2000 VL 20 IS 3 BP 1073 EP 1084 PG 12 WC Neurosciences SC Neurosciences & Neurology GA 278AG UT WOS:000084965300025 PM 10648713 ER PT J AU Sans, N Petralia, RS Wang, YX Blahos, J Hell, JW Wenthold, RJ AF Sans, N Petralia, RS Wang, YX Blahos, J Hell, JW Wenthold, RJ TI A developmental change in NMDA receptor-associated proteins at hippocampal synapses SO JOURNAL OF NEUROSCIENCE LA English DT Article DE MAGUKs; PSD-93; PSD-95; SAP-102; glutamate receptors; postsynaptic density; hippocampus ID TUMOR-SUPPRESSOR PROTEIN; NITRIC-OXIDE SYNTHASE; POSTSYNAPTIC DENSITY; IMMUNOCYTOCHEMICAL LOCALIZATION; GLUTAMATE RECEPTORS; GUANYLATE KINASES; MAMMALIAN RETINA; NERVOUS-SYSTEM; PSD-95 FAMILY; RAT-BRAIN AB The membrane-associated guanylate kinases [Chapsyn-110/postsynaptic density-93 (PSD-93), synapse-associated protein-90 (SAP-90)/PSD-95, and SAP-102] are believed to cluster and anchor NMDA receptors at the synapse and to play a role in signal transduction. We have investigated the developmental changes in expression of these proteins in rat hippocampus using biochemical analyses and quantitative immunogold electron microscopy. At postnatal day 2 (P2), SAP-102 was highly expressed, whereas PSD-93 and PSD-95 were low. SAP-102 expression increased during the first week, stayed stable through P35, and showed a reduced expression at 6 months. From P2 through 6 months, PSD-93 and PSD-95 increased. For PSD-95, the percent of labeled synapses increased almost threefold with age, whereas the number of gold particles per labeled synapse did not change significantly, suggesting that the increase in PSD-95 is attributable primarily to an increase in the number of synapses containing PSD-95. In contrast, for SAP-102, both percent labeled synapses and the number of gold particles per labeled synapse decreased during this time. From Western blots of hippocampus and immunogold analysis of CA1 synapses, the high expression of NR2B at P2 coincides with the high level of SAP-102 at synapses, whereas the later expression of NR2A coincides with that of PSD-93 and PSD-95. To determine whether the changes in PSD-93/95 and SAP-102 reflect preferred associations with NR2A and NR2B, respectively, we measured co-immunoprecipitation in the adult hippocampus. These studies suggest that there is a preference for complexes of NR2A/PSD-93/95 and NR2B/SAP-102. These results indicate that individual receptor-associated proteins may have specific functions that are critical to synapse development. C1 Natl Inst Deafness & Other Commun Disorders, Neurochem Lab, NIH, Bethesda, MD 20892 USA. Univ Wisconsin, Dept Pharmacol, Madison, WI 53706 USA. RP Sans, N (reprint author), Natl Inst Deafness & Other Commun Disorders, Neurochem Lab, NIH, Bldg 36,Room 5D08, Bethesda, MD 20892 USA. RI Blahos, Jaroslav/G-4397-2014 FU NINDS NIH HHS [NS 35563] NR 50 TC 337 Z9 345 U1 3 U2 7 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD FEB 1 PY 2000 VL 20 IS 3 BP 1260 EP 1271 PG 12 WC Neurosciences SC Neurosciences & Neurology GA 278AG UT WOS:000084965300042 PM 10648730 ER PT J AU Johnstone, LE Brown, CH Meeren, HKM Vuijst, CL Brooks, PJ Leng, G Russell, JA AF Johnstone, LE Brown, CH Meeren, HKM Vuijst, CL Brooks, PJ Leng, G Russell, JA TI Local morphine withdrawal increases c-fos gene, fos protein, and oxytocin gene expression in hypothalamic magnocellular neurosecretory cells SO JOURNAL OF NEUROSCIENCE LA English DT Article DE adrenoreceptors; benoxathian; dependence; hypothalamic paraventricular nucleus; microdialysis; naloxone; opioid; supraoptic nucleus ID IMMEDIATE-EARLY GENES; SUPRAOPTIC NUCLEUS; ANESTHETIZED RATS; OPIATE WITHDRAWAL; MESSENGER-RNA; IN-VIVO; NERVOUS-SYSTEM; ICV MORPHINE; NEURONS; ACTIVATION AB We measured stimulation of c-fos and oxytocin gene expression during excitation of oxytocin cells induced by systemic or local morphine withdrawal. Female rats were made morphine-dependent by intracerebroventricular morphine infusion over 5 d. Morphine withdrawal, induced by systemic injection of the opioid antagonist naloxone (5 mg/kg) in conscious or anesthetized rats, increased the density of c-fos messenger RNA and of oxytocin heterogeneous nuclear RNA in supraoptic nucleus cells compared with those of nonwithdrawn rats; c-fos messenger RNA was also increased in the magnocellular and parvocellular paraventricular nuclei of withdrawn rats. Morphine withdrawal increased the number of Fos-immunoreactive cells in the supraoptic and magnocellular paraventricular nuclei of conscious or pentobarbitone-anesthetized rats. Morphine withdrawal also increased Fos-immunoreactive cell numbers in the parvocellular paraventricular nucleus of conscious but not anesthetized rats. Central administration of the alpha(1)-adrenoreceptor antagonist benoxathian (5 mu g/min) did not prevent morphine withdrawal-induced increases in the numbers of Fos-immunoreactive neurons in the supraoptic or magnocellular paraventricular nucleus. Unilateral microdialysis administration of naloxone (10(-5) M) into the supraoptic nucleus of anesthetized morphine-dependent rats increased Fos-immunoreactive cell numbers compared with the contralateral nucleus. Finally, we investigated whether dependence could be induced by chronic unilateral infusion of morphine into a supraoptic nucleus; systemic naloxone (5 mg/kg) increased Fos-immunoreactive cell numbers in the morphine-infused nucleus compared with the contralateral nucleus. Thus, morphine withdrawal excitation increases c-fos and oxytocin gene expression in supraoptic nucleus neurons. This occurs independently from excitation of their ascending noradrenergic inputs, and both dependence and withdrawal can be induced within the supraoptic nucleus. C1 Univ Edinburgh, Sch Med, Dept Biomed Sci, Edinburgh EH8 9XD, Midlothian, Scotland. NIAAA, NIH, Rockville, MD 20852 USA. RP Russell, JA (reprint author), Univ Edinburgh, Sch Med, Dept Biomed Sci, Edinburgh EH8 9XD, Midlothian, Scotland. RI Leng, Gareth/C-6688-2011; Brown, Colin/A-6235-2011 OI Leng, Gareth/0000-0002-2388-8466; Brown, Colin/0000-0003-2305-846X NR 47 TC 33 Z9 33 U1 0 U2 2 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD FEB 1 PY 2000 VL 20 IS 3 BP 1272 EP 1280 PG 9 WC Neurosciences SC Neurosciences & Neurology GA 278AG UT WOS:000084965300043 PM 10648731 ER PT J AU Zhang, JH Simonds, WF AF Zhang, JH Simonds, WF TI Copurification of brain G-protein beta 5 with RGS6 and RGS7 SO JOURNAL OF NEUROSCIENCE LA English DT Article DE signal transduction; G-proteins; regulators of G-protein signaling; Igs; affinity purification; mass spectroscopy ID SUBUNIT; IDENTIFICATION; EXPRESSION; COMPLEX; RETINA; REGION; MEMBER; FAMILY AB A structurally divergent G-protein beta subunit expressed in brain and retina, G beta(5), exhibits functional specialization in its protein-protein interactions in vitro. In retina, Gb5 has been isolated in a soluble complex with regulator of G-protein signaling RGS7. The function and molecular associations of G beta(5) in brain are unknown. To identify tightly bound proteins associated with G beta(5) in the brain, it was immunoaffinity-purified from a nonionic detergent extract of washed mouse brain membranes using an antibody directed against its N terminus. Elution with cognate peptide revealed a broad band of 55 kDa that coeluted with G beta(5) on SDS-PAGE. The copurifying 55 kDa band was identified as a similar to 1:1 mixture of RGS6 and RGS7 by matrix-assisted laser desorption ionization mass spectroscopic analysis of tryptic peptides. G beta(5) and RGS7 could be reciprocally coimmunoprecipitated from unfractionated brain membrane extracts confirming the tight association of native proteins. In contrast, immunoblotting of the peptide eluate revealed no copurifying G alpha q/11, G alpha i1/2, G gamma 2, G gamma 3, or G gamma 7. These findings implicate RGS6 and RGS7 in the function of G beta(5) in the brain and suggest that a large fraction of membrane-targeted G beta 5 has no associated G gamma subunit and therefore functions outside the canonical framework of G beta gamma interactions. C1 NIDDK, Metab Dis Branch, Bethesda, MD 20892 USA. RP Simonds, WF (reprint author), NIDDK, Metab Dis Branch, Natl Inst Hlth, Bldg 10, Room 8C-101, 10 Ctr Dr MSC 1752, Bethesda, MD 20892 USA. NR 22 TC 45 Z9 46 U1 0 U2 0 PU SOC NEUROSCIENCE PI WASHINGTON PA 11 DUPONT CIRCLE, NW, STE 500, WASHINGTON, DC 20036 USA SN 0270-6474 J9 J NEUROSCI JI J. Neurosci. PD FEB 1 PY 2000 VL 20 IS 3 BP art. no. EP RC59 PG 5 WC Neurosciences SC Neurosciences & Neurology GA 278AG UT WOS:000084965300003 PM 10648734 ER PT J AU McKay, R AF McKay, R TI Stem cells and the cellular organization of the brain SO JOURNAL OF NEUROSCIENCE RESEARCH LA English DT Editorial Material ID DROSOPHILA AB The modern field of neuroscience emerged in the hands of neuroanatomists at the end of the last century. They recognized that information flows through arrays of cells and that the structure of the cells shapes the passage of information through the brain. In the middle of this century, the role of ion flux in the movement of information along axons was established. The electron microscope and the microelectrode helped to explain information transfer at the synapse, and study of the modulation of synaptic strength is currently a major area of neuroscience research. The origin of the many types of neuron seen by the early anatomists remains enigmatic, but the solution to this mystery is now emerging. The identification of stem cells and of the mechanisms that control their differentiation into distinct neuron types will contribute to a new understanding of both the origin of neuronal types and neuronal circuits. The current enthusiasm for stem cells is stimulated by interest from both the academic and nanacademic communities. These enthusiasts recognize a simple truth, that the cells are smart; they are the agents that control cell number and type in the brain. They create the structure that generates the higher order brain function. If these premises are true, then in the new century our understanding of the nervous system wilt be transformed by the facts of stem cell biology. Published 2000 Wiley-Liss, Inc.(dagger) C1 NINDS, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP McKay, R (reprint author), NINDS, Mol Biol Lab, NIH, 36 Convent Dr,Bldg 36,Room 5A29, Bethesda, MD 20892 USA. NR 15 TC 23 Z9 23 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0360-4012 J9 J NEUROSCI RES JI J. Neurosci. Res. PD FEB 1 PY 2000 VL 59 IS 3 BP 298 EP 300 DI 10.1002/(SICI)1097-4547(20000201)59:3<298::AID-JNR2>3.0.CO;2-U PG 3 WC Neurosciences SC Neurosciences & Neurology GA 280MX UT WOS:000085107500002 PM 10679764 ER PT J AU Molne, M Studer, L Tabar, V Ting, YT Eiden, MV McKay, RDG AF Molne, M Studer, L Tabar, V Ting, YT Eiden, MV McKay, RDG TI Early cortical precursors do not undergo LIF-mediated astrocytic differentiation SO JOURNAL OF NEUROSCIENCE RESEARCH LA English DT Article DE cell differentiation; CNS precursor cell; leukemia inhibitory factor; astrocyte; neuron ID CENTRAL-NERVOUS-SYSTEM; LEUKEMIA INHIBITORY FACTOR; NEUROEPITHELIAL STEM-CELLS; CEREBRAL-CORTEX; CELLULAR DIVERSITY; FACTOR-RECEPTOR; GROWTH-FACTOR; CNS; RAT; PROLIFERATION AB Multipotential stem cells have been isolated from the developing and adult CNS. Similar identified factors control the differentiation of these cells. A striking example is the instructive action of CNTF/LIF activating the JAW STAT pathway to induce astrocytic differentiation in both fetal and adult CNS stem cells. Here we show that E12 cortical precursors express functional LIF receptors but do not exhibit this differentiation response to CNTF/LIF either in explant or in dissociated cell culture. The lack of response to LIF-induced astrocytic differentiation is maintained in cocultures with LIF responsive cells derived from E15 cortex. This suggests cell intrinsic differences between early and late stage precursors in the interpretation of LIF-mediated signaling; however, the early nestin-positive precursor population differentiates into both neurons and neural crest derivatives. These data define differences between CNS stem cells from different stages of cortical development. Published 2000 Wiley-Liss, Inc.(dagger) C1 NINDS, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. NIMH, Unit Mol Virol, Lab Cell Mol Reg, Bethesda, MD 20892 USA. Mem Sloan Kettering Canc Ctr, Div Neurosurg, New York, NY 10021 USA. RP McKay, RDG (reprint author), NINDS, Mol Biol Lab, NIH, 36 Convent Dr,Bldg 36,Room 5A29, Bethesda, MD 20892 USA. NR 45 TC 70 Z9 78 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0360-4012 J9 J NEUROSCI RES JI J. Neurosci. Res. PD FEB 1 PY 2000 VL 59 IS 3 BP 301 EP 311 DI 10.1002/(SICI)1097-4547(20000201)59:3<301::AID-JNR3>3.0.CO;2-H PG 11 WC Neurosciences SC Neurosciences & Neurology GA 280MX UT WOS:000085107500003 PM 10679765 ER PT J AU Brenneman, DE AF Brenneman, DE TI Gordon Guroff - Obituary SO JOURNAL OF NEUROSCIENCE RESEARCH LA English DT Biographical-Item C1 NICHD, Sect Dev & Mol Pharmacol, Bethesda, MD USA. RP Brenneman, DE (reprint author), NICHD, Sect Dev & Mol Pharmacol, Bethesda, MD USA. NR 1 TC 0 Z9 0 U1 0 U2 0 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0360-4012 J9 J NEUROSCI RES JI J. Neurosci. Res. PD FEB 1 PY 2000 VL 59 IS 3 BP 464 EP 465 PG 2 WC Neurosciences SC Neurosciences & Neurology GA 280MX UT WOS:000085107500022 ER EF