FN Thomson Reuters Web of Science™ VR 1.0 PT J AU Adelstein, RS Takeda, K Conti, MA Golomb, E Kishi, H Preston, Y Yu, ZX Ferrans, VJ Wei, QZ AF Adelstein, RS Takeda, K Conti, MA Golomb, E Kishi, H Preston, Y Yu, ZX Ferrans, VJ Wei, QZ TI The A,B,C of nonmuscle myosin II SO JOURNAL OF GENERAL PHYSIOLOGY LA English DT Meeting Abstract C1 NHLBI, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 3 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 JUL PY 2001 VL 118 IS 1 MA 39 BP 18A EP 18A PG 1 WC Physiology SC Physiology GA 451CD UT WOS:000169782800050 ER PT J AU Wu, XF Rao, K Bowers, MB Copeland, NG Jenkins, NA Hammer, JA AF Wu, XF Rao, K Bowers, MB Copeland, NG Jenkins, NA Hammer, JA TI Rab27a enables myosin Va-dependent melanosome capture by recruiting the myosin to the organelle SO JOURNAL OF GENERAL PHYSIOLOGY LA English DT Meeting Abstract C1 NHLBI, NIH, Bethesda, MD 20892 USA. NCI, Frederick Canc Res & Dev Ctr, Mouse Canc Genet Program, Frederick, MD 21702 USA. NR 0 TC 0 Z9 0 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 JUL PY 2001 VL 118 IS 1 MA 52 BP 23A EP 24A PG 2 WC Physiology SC Physiology GA 451CD UT WOS:000169782800062 ER PT J AU Albrecht, MA Colegrove, SL Hongpaisan, J Pivovarova, NB Andrews, SB Friel, DD AF Albrecht, MA Colegrove, SL Hongpaisan, J Pivovarova, NB Andrews, SB Friel, DD TI Multiple modes of calcium-induced calcium release in sympathetic neurons I: Attenuation of endoplasmic reticulum Ca2+ accumulation at low [Ca2+](i) during weak depolarization SO JOURNAL OF GENERAL PHYSIOLOGY LA English DT Article DE calcium signaling; endoplasmic reticulum; caffeine; ryanodine; electron probe X-ray microanalysis ID CEREBELLAR PURKINJE-CELLS; RAT SENSORY NEURONS; SARCOPLASMIC-RETICULUM; RYANODINE RECEPTORS; CA-2+ TRANSIENTS; STORES; CAFFEINE; OSCILLATIONS; CHANNELS; ELEVATIONS AB Many cells express ryanodine receptors (RyRs) whose activation is thought to amplify depolarization-evoked elevations in cytoplasmic Ca2+ concentration ([Ca2+](i)) through a process of Ca2+-induced Ca2+ release (CICR). In neurons, it is usually assumed that CICR triggers net Ca2+ release From all ER Ca2+ store. However, since net ER Ca2+ transport depends oil the relative rates of Ca2+ uptake and release via distinct pathways, weak activation of a CICR pathway during periods of ER Ca accumulation would have a totally different effect: attenuation of Ca2+ accumulation. Stronger CICR activation at higher [Ca2+](i) could further attenuate Ca2+ accumulation or trigger net Ca2+ release, depending oil the quantitative properties of the underlying Ca2+ transporters. This and the companion study (Hongpaisan, J. N.B. Pivovarova, S.L. Colgrove, R.D. Leapman, and D.D. Friel, and S.B. Andrews. 2001. J. Gen. Physiol 118:101-112) investigate which of these CICR "modes" operate during depolarization-induced Ca-- entry in sympathetic neurons. The present study focuses oil small [Ca2+](i) elevations (less than similar to 350 nM) evoked by weak depolarization. The following two approaches were used: (1) Ca2+ fluxes were estimated from simultaneous measurements of [Ca2+], and I-Ca in fura-2-loaded cells (perforated patch conditions), and (2) total ER Ca concentrations ([Ca]FR) were measured using X-ray microanalysis. Flux analysis revealed triggered net Ca2+ release during depolarization ill the Presence but not the absence of caffeine, and [Ca2+](i) responses were accelerated by SERCA inhibitors, implicating ER Ca2+ accumulation, which was confirmed by direct [Ca]LR measurements. Ryanodine abolished caffeine-induced CICR and enhanced depolarization-induced ER Ca2+ accumulation, indicating that activation of the CICR pathway normally attenuates ER Ca2+ accumulation, which is a novel mechanism for accelerating evoked [Ca2+](i) responses. Theory shows how such a low gain mode of CICR can operate during weak stimulation and switch to net Ca2+ release at high [Ca2+](i), a transition demonstrated in the companion study. These results emphasize the importance of the relative rates of Ca2+ uptake and release in defining ER contributions to depolarization-induced Ca2+ signals. C1 Case Western Reserve Univ, Dept Neurosci, Cleveland, OH 44106 USA. Natl Inst Neurol Disorders & Stroke, Neurobiol Lab, NIH, Bethesda, MD 20892 USA. RP Friel, DD (reprint author), Case Western Reserve Univ, Dept Neurosci, 10900 Euclid Ave, Cleveland, OH 44106 USA. FU NINDS NIH HHS [NS-33514, R29 NS033514, R01 NS033514] NR 52 TC 34 Z9 34 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 JUL PY 2001 VL 118 IS 1 BP 83 EP 100 DI 10.1085/jgp.118.1.83 PG 18 WC Physiology SC Physiology GA 451CD UT WOS:000169782800008 PM 11429446 ER PT J AU Hongpaisan, J Pivovarova, NB Colegrove, SL Leapman, RD Friel, DD Andrews, SB AF Hongpaisan, J Pivovarova, NB Colegrove, SL Leapman, RD Friel, DD Andrews, SB TI Multiple modes of calcium-induced calcium release in sympathetic neurons II: A [Ca2+](i)- and location-dependent transition from endoplasmic reticulum Ca accumulation to net Ca release SO JOURNAL OF GENERAL PHYSIOLOGY LA English DT Article DE calcium signaling; mitochondria; ryanodine; electron probe X-ray microanalysis; electron energy loss spectrum imaging ID CEREBELLAR PURKINJE NEURONS; PERMEABILITY TRANSITION; HIPPOCAMPAL-NEURONS; PYRAMIDAL NEURONS; CHROMAFFIN CELLS; STORES; TRANSIENTS; DEPOLARIZATION; CAFFEINE; SEQUESTRATION AB CR front all intracellular store, he re directly characterized as the ER, usually refers to net Ca2+ release that amplifies evoked elevations in cytosolic free calcium ([Ca2+](i)). However, the companion paper (Albrecht, M.A., S.L. Colegrove, J. Hongpaisan, N.B. Pivovarova, S.B. Andrews. and D.D. Friel. 2001. J. Gen. Physiol. 118:83-100) shows that in sympathetic neurons, small elevations evoked by weak depolarization stimulate ER Ca accumulation, but at a rate attenuated by activation of a ryanodine-sensitive CICR pathway. Here, we have measured depolarization-evoked changes in total ER Ca concentration ([Ca](ER)) as a function of [Ca2+](i), and found that progressively larger [Ca2+](i) elevations cause a graded transition from ER Ca accumulation to net release, consistent with the expression of multiple modes of CICR. [Ca](ER) is relatively high at rest (12.8 +/- 0.9 mmol/kg dry weight, mean +/- SEM) and is reduced by thapsigargin or ryanodine (5.5 +/- 0.7 and 4.7 +/- 1.1 mmol/kg, respectively). [Ca](ER) rises during weak depolarization (to 17.0 +/- 1.6 mmol/kg over 120s, [Ca2+](i) less than similar to 350 nM), changes little in response to stronger depolarization (12.1 +/- 1.1 mmol/kg, [Ca2+], similar to 700 nM), and declines (to 6.5 +/- 1.0 mmol/kg) with larger [Ca2+](i) elevations (>1 muM) evoked by the same depolarization when mitochondrial Ca2+ uptake is inhibited (FCCP). Thus, net ER Ca2+ transport exhibits a biphasic dependence on [Ca2+](i). With mitochondrial Ca2+ uptake enabled, [Ca](ER) rises after repolarization (to 16.6 +/- 1.8 mmol/kg at 15 min) as [Ca2+](i) falls within the permissive range for ER Ca accumulation over a period lengthened by mitochondrial Ca2+ release. Finally, although spatially averaged [Ca](ER) is unchanged during strong depolarization, net ER Ca2+ release still occurs, but only in the outermost similar to5-mum cytoplasmic shell where [Ca2+], should reach its highest levels. Since mitochondrial Ca accumulation occurs preferentially in peripheral cytoplasm, as demonstrated here by electron energy loss Ca maps, the Ca content of ER and mitochondria exhibit reciprocal dependencies on proximity to sites of Ca2+ entry, possibly reflecting indirect mitochondrial regulation of ER Ca2+ transport. C1 Natl Inst Neurol Disorders & Stroke, Neurobiol Lab, NIH, Bethesda, MD 20892 USA. NIH, Bioengn & Phys Sci Program, Off Director, Bethesda, MD 20892 USA. Case Western Reserve Univ, Dept Neurosci, Cleveland, OH 44106 USA. RP Andrews, SB (reprint author), Natl Inst Neurol Disorders & Stroke, Neurobiol Lab, NIH, Bldg 36,Room 2A-21,36 Convent Dr, Bethesda, MD 20892 USA. FU NINDS NIH HHS [NS-33514, R01 NS033514, R29 NS033514] NR 56 TC 34 Z9 34 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 JUL PY 2001 VL 118 IS 1 BP 101 EP 112 DI 10.1085/jgp.118.1.101 PG 12 WC Physiology SC Physiology GA 451CD UT WOS:000169782800009 PM 11429447 ER PT J AU Kabrane-Lazizi, Y Zhang, MD Purcell, RH Miller, KD Davey, RT Emerson, SU AF Kabrane-Lazizi, Y Zhang, MD Purcell, RH Miller, KD Davey, RT Emerson, SU TI Acute hepatitis caused by a novel strain of hepatitis E virus most closely related to United States strains SO JOURNAL OF GENERAL VIROLOGY LA English DT Article ID OPEN-READING FRAME-2; MOLECULAR-CLONING; BLOOD-DONORS; INSECT CELLS; E INFECTION; IDENTIFICATION; HEV; ANTIBODY; SWINE; SEROREACTIVITY AB A unique hepatitis E virus (HEV) strain was identified as the aetiological agent of acute hepatitis in a United States (US) patient who had recently returned from vacation in Thailand, a country in which HEV is endemic. Sequence comparison showed that this HEV strain was most similar, but not identical, to the swine and human HEV strains recovered in the US. Phylogenetic analysis revealed that this new HEV isolate was closer to genotype 3 strains than to the genotype 1 strains common in Asia. The fact that this HEV was closely related to strains recovered in countries where HEV is not endemic and was highly divergent from Asian HEV strains raises the questions of where the patient's infection was acquired and of whether strains are geographically as localized as once thought. C1 NIAID, Infect Dis Lab, Hepatitis Viruses Sect, NIH, Bethesda, MD 20892 USA. NIAID, Infect Dis Lab, Mol Hepatitis Sect, NIH, Bethesda, MD 20892 USA. NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. NIH, Ctr Clin, Dept Crit Care Med, Bethesda, MD 20892 USA. RP Emerson, SU (reprint author), NIAID, Infect Dis Lab, Hepatitis Viruses Sect, NIH, Bethesda, MD 20892 USA. NR 35 TC 32 Z9 34 U1 0 U2 0 PU SOC GENERAL MICROBIOLOGY PI READING PA MARLBOROUGH HOUSE, BASINGSTOKE RD, SPENCERS WOODS, READING RG7 1AE, BERKS, ENGLAND SN 0022-1317 J9 J GEN VIROL JI J. Gen. Virol. PD JUL PY 2001 VL 82 BP 1687 EP 1693 PN 7 PG 7 WC Biotechnology & Applied Microbiology; Virology SC Biotechnology & Applied Microbiology; Virology GA 445AU UT WOS:000169435800018 PM 11413380 ER PT J AU Lee, YJ Shacter, E AF Lee, YJ Shacter, E TI Fas aggregation does not correlate with Fas-mediated apoptosis SO JOURNAL OF IMMUNOLOGY LA English DT Article ID PROTEIN-KINASE-C; INDUCED CELL-DEATH; AUTOIMMUNE LYMPHOPROLIFERATIVE SYNDROME; SIGNALING COMPLEX DISC; JURKAT T-CELLS; LYMPHOMA-CELLS; TNF RECEPTOR; CD95 FAS/APO-1; ACTIVATION; LYMPHOCYTES AB Cross-linking of cell surface Fas molecules by Fas ligand or by agonistic anti-Fas Abs induces cell death by apoptosis. We found that a serine protease inhibitor, N-tosyl-L-lysine chloromethyl ketone (TLCK), dramatically enhances Fas-mediated apoptosis in the human T cell line Jurkat and in various B cell lines resistant to Fas-mediated apoptosis. The enhancing effect of TLCK is specific to Fas-induced cell death, with no effect seen on TNF-alpha or TNF-related apoptosis-inducing ligand-induced apoptosis. TLCK treatment had no effect on Fas expression levels on the cell surface, and neither promoted death-inducing signaling complex formation nor decreased expression levels of cellular inhibitors of apoptosis (FLICE inhibitory protein, X chromosome-linked inhibitor of apoptosis, and Bcl-2). Activation of the Fas-mediated apoptotic pathway by anti-Fas Ab is accompanied by aggregation of Fas molecules to form oligomers that are stable to boiling in SDS and beta -NM. Fas aggregation is often considered to be required for Fas-mediated apoptosis. However, sensitization of cells to Fas-mediated apoptosis by TLCK or other agents (cycloheximide, protein kinase C inhibitors) causes less Fas aggregation during the apoptotic process compared with that in nonsensitized cells. These results show that Fas aggregation and Fas-mediated apoptosis are not directly correlated and may even be inversely correlated. C1 Ctr Biol & Evaluat & Res, Div Therapeut Prot, Immunol Lab, Food & Drug Adm, Bethesda, MD 20892 USA. RP Lee, YJ (reprint author), NINDS, Surg Neurol Branch, Biochem Sect, NIH, Bldg 10,Room 4N258, Bethesda, MD 20892 USA. NR 48 TC 24 Z9 24 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 JUL 1 PY 2001 VL 167 IS 1 BP 82 EP 89 PG 8 WC Immunology SC Immunology GA 471VJ UT WOS:000170949100013 PM 11418635 ER PT J AU Cherukuri, A Cheng, PC Pierce, SK AF Cherukuri, A Cheng, PC Pierce, SK TI The role of the CD19/CD21 complex in B cell processing and presentation of complement-tagged antigens SO JOURNAL OF IMMUNOLOGY LA English DT Article ID HUMORAL IMMUNE-RESPONSE; T-DEPENDENT ANTIGEN; CROSS-LINKING; CLASS-II; ACQUIRED-IMMUNITY; RECEPTOR; LYMPHOCYTES; EXPRESSION; MICE; CD19 AB The CD19/CD21 complex is an essential B cell coreceptor that functions synergistically to enhance signaling through the B cell Ag receptor in response to T cell-dependent, complement-tagged Ags. In this study, we use a recombinant protein containing three tandemly arranged copies of Od and the Ag hen egg lysozyme, shown to be a highly effective immunogen in vivo, to evaluate the role of the CD19/CD21 complex in Ag processing in B cells. Evidence is provided that coengagement of the CD19/CD21 complex results in more rapid and efficient production of antigenic peptide/class II complexes as compared with B cell Ag receptor-mediated processing alone. The CD19/CD21 complex does not itself target complement-tagged Ags for processing, but rather appears to influence B cell Ag processing through its signaling function. The ability of the CD19/CD21 complex to augment processing may be an important element of the mechanism by which the CD19/CD21 complex functions to promote B cell responses to T cell-dependent complement-tagged Ags in vivo. C1 NIAID, Immunogenet Lab, NIH, Rockville, MD 20852 USA. Northwestern Univ, Dept Biochem Mol Biol & Cell Biol, Evanston, IL 60208 USA. RP Pierce, SK (reprint author), NIAID, Immunogenet Lab, NIH, Twinbrook 2,12441 Parklawn Dr,Room 200B,MSC 8180, Rockville, MD 20852 USA. NR 36 TC 109 Z9 120 U1 3 U2 5 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD JUL 1 PY 2001 VL 167 IS 1 BP 163 EP 172 PG 10 WC Immunology SC Immunology GA 471VJ UT WOS:000170949100023 PM 11418645 ER PT J AU Su, SB Silver, PB Zhang, MF Chan, CC Caspi, RR AF Su, SB Silver, PB Zhang, MF Chan, CC Caspi, RR TI Pertussis toxin inhibits induction of tissue-specific autoimmune disease by disrupting G protein-coupled signals SO JOURNAL OF IMMUNOLOGY LA English DT Article ID EXPERIMENTAL ALLERGIC ENCEPHALOMYELITIS; BORDETELLA-PERTUSSIS; IMMUNE-RESPONSES; B-OLIGOMER; MICE; UVEORETINITIS; SUSCEPTIBILITY; LYMPHOCYTE; ACTIVATION; CELLS AB Pertussis toxin (PTX) has been used for many years as an adjuvant that promotes development of tissue-specific experimental autoimmune diseases such as experimental autoimmune encephalomyelitis, experimental autoimmune uveitis (EAU), and others. Enhancement of vascular permeability and of Th1 responses have been implicated in this effect. Here we report a surprising observation that, in a primed system, PTX can completely block the development of EAU. Disease was induced in B10.RIII mice by adoptive transfer of uveitogenic T cells, or by immunization with a uveitogenic peptide. A single injection of PTX concurrently with infusion of the uveitogenic T cells, or two injections 7 and 10 days after active immunization, completely blocked development of EAU. EAU also was prevented by a 1-h incubation in vitro of the uveitogenic T cells with PTX before infusing them into recipients. Uveitogenic T cells treated with PTX in vitro and lymphoid cells from mice treated with PTX in vivo failed to migrate to chemokines in a standard chemotaxis assay. Neither the isolated B-oligomer subunit of PTX that lacks ADP ribosyltransferase activity nor the related cholera toxin that ADP-ribosylates G(s) (but not G(i)) proteins blocked EAU induction or migration to chemokines. We conclude that PTX present at the time of cell migration to the target organ prevents EAU, and propose that it does so at least in part by disrupting signaling through G(i) protein-coupled receptors. Thus, the net effect of PTX on autoimmune disease would represent an integration of enhancing and inhibitory effects. C1 NEI, Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Caspi, RR (reprint author), NEI, Immunol Lab, NIH, 10 Ctr Dr,10-10N222, Bethesda, MD 20892 USA. NR 45 TC 25 Z9 28 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 JUL 1 PY 2001 VL 167 IS 1 BP 250 EP 256 PG 7 WC Immunology SC Immunology GA 471VJ UT WOS:000170949100034 PM 11418656 ER PT J AU Wang, JH Shelly, L Miele, L Boykins, R Norcross, MA Guan, E AF Wang, JH Shelly, L Miele, L Boykins, R Norcross, MA Guan, E TI Human notch-1 inhibits NF-kappa B activity in the nucleus through a direct interaction involving a novel domain SO JOURNAL OF IMMUNOLOGY LA English DT Article ID T-CELL DEVELOPMENT; TRANSCRIPTION FACTOR; SIGNALING PATHWAY; NEOPLASTIC LESIONS; ANKYRIN REPEATS; HUMAN HOMOLOG; DNA-BINDING; DROSOPHILA; EXPRESSION; RECEPTOR AB Notch participates in diverse cell fate decisions throughout embryonic development and postnatal life. Members of the NF-kappaB/Rel family of transcription factors are involved in the regulation of a variety of genes important for immune function. The biological activity of the NF-kappaB transcription factors is controlled by I kappaB proteins. Our previous work demonstrated that an intracellular, constitutively active form of human Notch-1/translocation-associated Notch homologue-1 (Notch(IC)) functions as an I kappaB molecule with specificity for the NF-kappaB p50 subunit and physically interacts with NF-kappaB in T cells. In the current study, we investigated the roles of different domains of Notch(IC) in the regulation of NF-kappaB-directed gene expression and NF-kappab DNA binding activity. We found that Notch(IC) localizes to the nucleus and that a region in the N-terminal portion of Notch(IC), not the six ankyrin repeats, is responsible for the inhibitory effects of Notch on NF-kappaB-directed gene expression and NF-kappaB DNA binding activity. The N-terminal portion of Notch(IC) inhibited p50 DNA binding and interacted specifically with p50 subunit, not p65 of NF-kappaB. The interaction between Notch and NF-kappaB indicates that in addition to its role in the development of the immune system, Notch-1 may also have critical functions in the immune response, inflammation, viral infection, and apoptosis through control of NF-kappaB-mediated gene expression. C1 NIH, US FDA, Ctr Biol Evaluat & Res, Div Therapeut Prot,Lab Gene Regulat, Bethesda, MD 20892 USA. RP Wang, JH (reprint author), NIH, US FDA, Ctr Biol Evaluat & Res, Div Therapeut Prot,Lab Gene Regulat, Bldg 29B,Room 4E12,HFM-541,8800 Rockville Pike, Bethesda, MD 20892 USA. FU NCI NIH HHS [1R01CA84065-01] NR 72 TC 81 Z9 85 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 JUL 1 PY 2001 VL 167 IS 1 BP 289 EP 295 PG 7 WC Immunology SC Immunology GA 471VJ UT WOS:000170949100040 PM 11418662 ER PT J AU Dong, YS Tang, LP Letterio, JJ Benveniste, EN AF Dong, YS Tang, LP Letterio, JJ Benveniste, EN TI The Smad3 protein is involved in TGF-beta inhibition of class II transactivator and class II MHC expression SO JOURNAL OF IMMUNOLOGY LA English DT Article ID GROWTH-FACTOR-BETA; IFN-GAMMA INDUCTION; GENE-EXPRESSION; TARGETED DISRUPTION; IMMUNE-RESPONSES; BINDING-PROTEIN; TNF-ALPHA; CIITA; STAT1; ASTROCYTES AB TGF-beta is a immunoregulatory cytokine that inhibits class ii MHC expression in a variety of cell types. Previous studies have shown that the class II MHC transactivator (CIITA), a master regulator that controls class II MHC expression, is targeted by TGF-beta for repression of IFN-gamma -induced class II MHC expression in astrocytes. The mechanism(s) underlying the TGF-beta inhibitory effect is not understood. In this study, we demonstrate that TGF-beta inhibition of CIITA expression occurs at the transcriptional level, and that both constitutive and IFN-gamma -induced human CIITA type IV promoter activity is inhibited by TGF-beta. TGF-beta does not affect the signaling events that mediate IFN-gamma activation of CIITA expression; i.e, TGF-beta does not inhibit IFN-gamma -induced STAT-la phosphorylation and/or DNA binding ability, nor is IFN-gamma induction of IFN regulatory factor affected. The inhibitory effect of TGF-beta on the type IV CIITA promoter is mediated through a promoter region within 80 bp from the transcription start site. Elimination of TGF-beta inhibition of class II MHC and CIITA expression in Smad3-deficient astrocytes, as well as restoration of the inhibitory effect by overexpression of the Smad3 protein, demonstrates that Smad3 is essential in mediating TGF-beta inhibition of CIITA and class II MHC expression. C1 Univ Alabama, Dept Cell Biol, Birmingham, AL 35294 USA. NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bethesda, MD 20892 USA. RP Benveniste, EN (reprint author), Univ Alabama, Dept Cell Biol, 1918 Univ Blvd,MCLM 395, Birmingham, AL 35294 USA. FU NIADDK NIH HHS [AM20614]; NINDS NIH HHS [NS36765] NR 49 TC 53 Z9 59 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 JUL 1 PY 2001 VL 167 IS 1 BP 311 EP 319 PG 9 WC Immunology SC Immunology GA 471VJ UT WOS:000170949100043 PM 11418665 ER PT J AU Koniaris, LG Zimmers-Koniaris, T Hsiao, EC Chavin, K Sitzmann, JV Farber, JM AF Koniaris, LG Zimmers-Koniaris, T Hsiao, EC Chavin, K Sitzmann, JV Farber, JM TI Cytokine-responsive gene-2/IFN-inducible protein-10 expression in multiple models of liver and bile duct injury suggests a role in tissue regeneration SO JOURNAL OF IMMUNOLOGY LA English DT Article ID INTERFERON-INDUCIBLE PROTEIN-10; HEPATOCYTE GROWTH-FACTOR; TUMOR-NECROSIS-FACTOR; CHEMOKINE RECEPTOR CXCR4; PARTIAL-HEPATECTOMY; FACTOR-ALPHA; IN-VIVO; T-CELLS; GENE-EXPRESSION; MICE LACKING AB IFN-inducible protein-10 (IP-IO/CXCL10) is a CXC chemokine that targets both T cells and NK cells. Elevation of IP-10 expression has been demonstrated in a number of human diseases, including chronic cirrhosis and biliary atresia. Cytokine-responsive gene-2 (Crg-2), the murine ortholog of IP-10, was induced following CCl4 treatment of the hepatocyte-like cell line AML-12. Crg-2 expression was noted in vivo in multiple models of hepatic and bile duct injury, including bile duct ligation and CCl4, D-galactosamine, and methylene dianiline toxic liver injuries. Induction of Crg-2 was also examined following two-thirds hepatectomy, a model that minimally injures the remaining liver, but that requires a large hepatic regenerative response. Crg-2 was induced in a biphasic fashion after two-thirds hepatectomy, preceding each known peak of hepatocyte DNA synthesis. Induction of Crg-2 was also observed in the kidney, gut, thymus, and spleen within 1 h of two-thirds hepatectomy. Characteristic of an immediate early gene, pretreatment of mice with the protein synthesis inhibitor cycloheximide before either two-thirds hepatectomy or CCl4 injection led to Crg-2 superinduction. rIP-10 was demonstrated to have hepatocyte growth factor-inducing activity in vitro, but alone had no direct mitogenic effect on hepatocytes. Our data demonstrate that induction of Crg-2 occurs in several distinct models of liver injury and regeneration, and suggest a role for CRG-2/IP-10 in these processes. The Journal of Immunology, 2001. C1 Johns Hopkins Univ, Sch Med, Dept Mol Biol & Genet, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Dept Surg, Baltimore, MD 21205 USA. Med Univ S Carolina, Dept Surg, Charleston, SC 29425 USA. Univ Rochester, Sch Med, Dept Surg, Rochester, NY 14642 USA. NIAID, Clin Invest Lab, Bethesda, MD 20892 USA. RP Koniaris, LG (reprint author), Univ Rochester, Sch Med & Dent, Dept Surg, 601 Elmwood Ave, Rochester, NY 14642 USA. OI Zimmers, Teresa/0000-0001-7872-0540 FU NCI NIH HHS [5 T32 CA09139]; NIGMS NIH HHS [5 T32 GM07309] NR 62 TC 58 Z9 60 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 JUL 1 PY 2001 VL 167 IS 1 BP 399 EP 406 PG 8 WC Immunology SC Immunology GA 471VJ UT WOS:000170949100054 PM 11418676 ER PT J AU Schwarz, M Murphy, PM AF Schwarz, M Murphy, PM TI Kaposi's sarcoma-associated herpesvirus G protein-coupled receptor constitutively activates NF-kappa B and induces proinflammatory cytokine via a C-terminal signaling and chemokine production determinant SO JOURNAL OF IMMUNOLOGY LA English DT Article ID FIBROBLAST GROWTH-FACTOR; MAMMALIAN-CELLS; SPINDLE CELLS; EXPRESSION; INHIBITION; AIDS; HUMAN-HERPESVIRUS-8; RESPONSES; ONCOGENE; CULTURE AB Kaposi's sarcoma-associated herpesvirus (KSHV) is believed to be the causative agent of Kaposi's sarcoma (KS), a multicentric growth factor-dependent tumor common in AIDS patients characterized histopathologically by spindle cell proliferation, angiogenesis, and leukocyte infiltration. Recently, open reading frame 74 of KSHV has been implicated as a major viral determinant of KS. Open reading frame 74 encodes KSHV G protein-coupled receptor (GPCR), a constitutively active chemokine receptor that directly transforms NIH 3T3 cells in vitro and induces multifocal KS-like lesions in KSHV-GPCR-transgenic mice. Interestingly, receptor-positive cells are very rare in lesions from these mice, implicating an indirect mechanism of tumorigenesis. In this regard, here we report that expression of KSHV-GPCR in transfected epithelial, monocytic, and T cell lines induced constitutive activation of the immunoregulatory transcription factors AP-I and NF-kappaB. This was associated with constitutive induction of the proinflammatory NF-kappaB-dependent cytokines IL-1 beta, IL-6, and TNF-alpha, and chemokines monocyte chemoattractant protein-1 and IL-8, as well as the AP-1-dependent basic fibroblast growth factor. In addition, IL-2 and IL-4 production was induced in transfected Jurkat T cells. Truncation of the final five amino acids in the cytoplasmic tail of KSHV-GPCR caused complete loss of its transforming and NF-kappaB-inducing activities, without affecting receptor expression or ligand binding. These data suggest that KS results in part from KSHV-GPCR induction of proinflammatory cytokine and growth factor gene expression, mediated by a signaling determinant within the last five amino acids of the C terminus, a domain that is also critical for direct cell transformation. The Journal of Immunology, 2001. C1 NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. RP Murphy, PM (reprint author), NIAID, Host Def Lab, NIH, Bldg 10,Room 11N113, Bethesda, MD 20892 USA. NR 43 TC 133 Z9 138 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 JUL 1 PY 2001 VL 167 IS 1 BP 505 EP 513 PG 9 WC Immunology SC Immunology GA 471VJ UT WOS:000170949100067 PM 11418689 ER PT J AU Mackall, CL AF Mackall, CL TI Spreading the wealth: Antigen discovery in adult tumors can help hone the search for pediatric tumor antigens SO JOURNAL OF IMMUNOTHERAPY LA English DT Editorial Material ID T-LYMPHOCYTES; NEUROBLASTOMA; EXPRESSION; MAGE-1; CELLS; MELANOMA; PROTEIN C1 NCI, Pediat Oncol Branch, Bethesda, MD 20892 USA. RP Mackall, CL (reprint author), NCI, Pediat Oncol Branch, Bethesda, MD 20892 USA. NR 16 TC 1 Z9 1 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1053-8550 J9 J IMMUNOTHER JI J. Immunother. PD JUL-AUG PY 2001 VL 24 IS 4 BP 281 EP 282 DI 10.1097/00002371-200107000-00001 PG 2 WC Oncology; Immunology; Medicine, Research & Experimental SC Oncology; Immunology; Research & Experimental Medicine GA 469WU UT WOS:000170839700001 PM 11565827 ER PT J AU Alexander, HR AF Alexander, HR TI The effects of limb perfusion with tumor necrosis factor on circulating levels of proinflammatory cytokines SO JOURNAL OF IMMUNOTHERAPY LA English DT Editorial Material ID SYSTEMIC LEAKAGE C1 NCI, Surg Metab Sect, Surg Branch, Div Clin Sci, Bethesda, MD 20892 USA. RP Alexander, HR (reprint author), NCI, Surg Metab Sect, Surg Branch, Div Clin Sci, Bethesda, MD 20892 USA. NR 9 TC 1 Z9 1 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1053-8550 J9 J IMMUNOTHER JI J. Immunother. PD JUL-AUG PY 2001 VL 24 IS 4 BP 285 EP 286 DI 10.1097/00002371-200107000-00003 PG 2 WC Oncology; Immunology; Medicine, Research & Experimental SC Oncology; Immunology; Research & Experimental Medicine GA 469WU UT WOS:000170839700003 PM 11565829 ER PT J AU Schwartzentruber, DJ AF Schwartzentruber, DJ TI Guidelines for the safe administration of high-dose interleukin-2 SO JOURNAL OF IMMUNOTHERAPY LA English DT Review DE high-dose IL-2; toxicity; melanoma; renal cell carcinoma ID 199 CONSECUTIVE PATIENTS; RENAL-CELL CARCINOMA; METASTATIC MELANOMA; CANCER; TOXICITY AB High-dose interleukin-2 (IL-2) results in objective clinical regression of metastatic cancer in 15% to 17% of patients with melanoma and renal cell carcinoma. Durable complete regression of all metastases is seen in 6% to 8% of patients. Based on these findings, the U.S. Food and Drug Administration has approved the use of high-dose IL-2 for the treatment of patients with metastatic melanoma and renal cell carcinoma. Interleukin-2 administration is associated with many different side effects, and after many years of use, clinicians have learned how to safely administer high-dose IL-2. This article details practical guidelines for the safe administration of high-dose IL-2. C1 NCI, Surg Branch, NIH, Bethesda, MD 20892 USA. RP Schwartzentruber, DJ (reprint author), NCI, Surg Branch, NIH, Bldg 10,Room 2B0b, Bethesda, MD 20892 USA. NR 15 TC 74 Z9 78 U1 1 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1053-8550 J9 J IMMUNOTHER JI J. Immunother. PD JUL-AUG PY 2001 VL 24 IS 4 BP 287 EP 293 DI 10.1097/00002371-200107000-00004 PG 7 WC Oncology; Immunology; Medicine, Research & Experimental SC Oncology; Immunology; Research & Experimental Medicine GA 469WU UT WOS:000170839700004 PM 11565830 ER PT J AU Harada, M Li, YF El-Gamil, M Ohnmacht, GA Rosenberg, SA Robbins, PF AF Harada, M Li, YF El-Gamil, M Ohnmacht, GA Rosenberg, SA Robbins, PF TI Melanoma-reactive CD8(+) T cells recognize a novel tumor antigen expressed in a wide variety of tumor types SO JOURNAL OF IMMUNOTHERAPY LA English DT Article DE human; T lymphocytes; tumor immunity; melanoma ID OPEN READING FRAME; INFILTRATING LYMPHOCYTES; NEUROECTODERMAL TUMORS; INTRON SEQUENCE; GENE; IDENTIFICATION; TRANSLATION; CLONING; ENCODES; SYSTEM AB An autologous melanoma cell line selected for loss of expression of the immunodominant MART-1 and gp100 antigens was initially used to carry out a mixed lymphocyte tumor culture (MLTC) in a patient who expressed the human leukocyte antigen (HLA)-A1 and HLA-A2 class I major histocompatibility complex alleles. Ten clones identified from this MLTC seemed to recognize melanoma in an HLA-A1-restricted manner but failed to recognize a panel of previously described melanoma antigens. The screening of an autologous melanoma cDNA library with one HLA-A1-restricted melanoma-reactive T-cell clone resulted in the isolation of a cDNA clone called AIM-2 (antigen isolated from immunoselected melanoma-2). The AIM-2 transcript seemed to have retained an intronic sequence based on its alignment with genomic sequences as well as expressed sequence tags. This transcript was not readily detected after Northern blot analysis of melanoma mRNA, indicating that only low levels of this product may be expressed in tumor cells. Quantitative reverse transcriptase-polymerase chain reaction analysis, however, demonstrated a correlation between T-cell recognition and expression in HLA-A1-expressing tumor cell lines. A peptide that was encoded within a short open reading frame of 23 amino acids and conformed to the HLA-A1 binding motif RSDSGQQARY was found to represent the T-cell epitope. The AIM-2-reactive T-cell clone recognized a number of neuroectodermal tumors as well as breast, ovarian, and colon carcinomas that expressed HLA-A1, indicating that this represents a widely expressed tumor antigen. Thus, AIM-2 may represent a potential target for the development of vaccines in patients bearing tumors of a variety of histologies. C1 NCI, Surg Branch, NIH, Bethesda, MD 20892 USA. Kurume Univ, Sch Med, Fukuoka, Japan. Univ Texas, Sch Med, Houston, TX USA. RP Robbins, PF (reprint author), NCI, Surg Branch, NIH, Bldg 10,Room 2B42,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 24 TC 18 Z9 18 U1 0 U2 2 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1053-8550 J9 J IMMUNOTHER JI J. Immunother. PD JUL-AUG PY 2001 VL 24 IS 4 BP 323 EP 333 DI 10.1097/00002371-200107000-00008 PG 11 WC Oncology; Immunology; Medicine, Research & Experimental SC Oncology; Immunology; Research & Experimental Medicine GA 469WU UT WOS:000170839700008 PM 11565834 ER PT J AU Dudley, ME Wunderlich, J Nishimura, MI Yu, D Yang, JC Topalian, SL Schwartzentruber, DJ Hwu, P Marincola, FM Sherry, R Leitman, SF Rosenberg, SA AF Dudley, ME Wunderlich, J Nishimura, MI Yu, D Yang, JC Topalian, SL Schwartzentruber, DJ Hwu, P Marincola, FM Sherry, R Leitman, SF Rosenberg, SA TI Adoptive transfer of cloned melanoma-reactive T lymphocytes for the treatment of patients with metastatic melanoma SO JOURNAL OF IMMUNOTHERAPY LA English DT Article DE immunotherapy; melanoma; gp100; interleukin-2; T-cell clone ID TUMOR-INFILTRATING LYMPHOCYTES; CELL CLONES; INTERFERON-GAMMA; IN-VITRO; CD8(+); IMMUNOTHERAPY; VIRUS; INTERLEUKIN-2; EFFICACY; SURVIVAL AB This report describes a phase I study of the adoptive transfer of cloned melanoma antigen-specific T lymphocytes for therapy of patients with advanced melanoma. Clones were derived from peripheral blood lymphocytes or tumor-infiltrating lymphocytes of patients who had received prior immunization with the melanoma-associated antigen, gp100. In response to its cognate antigen, each clone used for treatment secreted large amounts of interferon-gamma and granulocyte-macrophage colony-stimulating factor, lesser amounts of interleukin (IL)-2 and tumor necrosis factor-alpha, and little or no IL-4 and IL-10. Clones also demonstrated recognition of human leukocyte antigen-matched melanomas using cytokine secretion and lysis assays. Twelve patients received 2 cycles of cells alone; I I patients received additional cycles of cells and were randomized between two schedules of IL-2 (125,000 IU/kg subcutaneously daily for 12 days versus 720,000 IU/kg intravenously every 8 h for 4 days). A total of 51 cycles of cells were administered, with an average of 1 x 10(10) cells per cycle. Peripheral blood samples were analyzed for persistence of transferred cells by T-cell receptor-specific polymerase chain reaction. Transferred cells reached a maximum level at I It after transfer but rapidly declined to undetectable levels by 2 weeks. One minor response and one mixed response were observed (both in the high-dose IL-2 arm). This report demonstrates the safety and feasibility of cloned T-cell transfer as a therapy for patients with cancer. The lack of clinical effectiveness of this protocol suggests that transfer of different or additional cell types or that modulation of the recipient host environment is required for successful therapy. C1 NCI, Surg Branch, NIH, Bethesda, MD 20892 USA. NIH, Dept Transfus Med, Bethesda, MD 20892 USA. Univ Chicago, Dept Surg, Sect Gen Surg, Chicago, IL 60637 USA. RP Dudley, ME (reprint author), NCI, Surg Branch, NIH, Bldg 10,Room 2B-08,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 21 TC 259 Z9 262 U1 0 U2 4 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1053-8550 J9 J IMMUNOTHER JI J. Immunother. PD JUL-AUG PY 2001 VL 24 IS 4 BP 363 EP 373 DI 10.1097/00002371-200107000-00012 PG 11 WC Oncology; Immunology; Medicine, Research & Experimental SC Oncology; Immunology; Research & Experimental Medicine GA 469WU UT WOS:000170839700012 PM 11565838 ER PT J AU Sconocchia, G Cococcetta, NY Campagnano, L Amadori, S Iorio, B Boffo, V Ferdinandi, V Del Principe, I Adorno, D Casciani, CU AF Sconocchia, G Cococcetta, NY Campagnano, L Amadori, S Iorio, B Boffo, V Ferdinandi, V Del Principe, I Adorno, D Casciani, CU TI Subcutaneous administration of interleukin-2 triggers Fc gamma receptor I expression on human peripheral blood neutrophils in solid and hematologic malignancies SO JOURNAL OF IMMUNOTHERAPY LA English DT Article DE IL-2; immunotherapy; Fc gamma RI; MNs; malignancies ID HUMAN POLYMORPHONUCLEAR LEUKOCYTES; COLONY-STIMULATING FACTOR; NECROSIS-FACTOR-ALPHA; RENAL-CELL CARCINOMA; INTERFERON-ALPHA; MYELOID CELLS; SERUM LEVELS; IGG; IL-2; GRANULOCYTES AB Freshly isolated human polymorphonuclear cells (PMNCs) constitutively express Fc gamma receptor (Fc gammaR) II and Fc gamma RIII on the cell surface but not Fc gamma RI. Cytokines such as interferon-gamma (IFN-gamma), granulocyte-macrophage colony-stimulating factor (CSF), and granulocyte-CSF trigger Fc gamma RI expression on (PMNCs). Because PMNCs express interleukin (IL)-2 receptor, we investigated whether IL-2 can induce Fc gamma RI expression on PMNCs isolated from IL-2-treated metastatic renal cell carcinoma (MRCC) and low-grade non-Hodgkin lymphoma (LGNHL) patients. Pretherapy flow cytometry analysis of Fc receptors on PMNCs did not show Fc gamma RI expression. Interestingly, 3 days after therapy, PMNCs displayed a detectable amount of Fc gamma RI on the cell surface. Kinetic studies on the in vivo effects of IL-2 on MRCC patients showed that Fc gamma RI was transiently expressed, starting within 3-6 days of therapy, remaining expressed for 10-15 days, and rapidly declining, whereas such expression remained stable for months in LGNHL patients. In contrast, Fc gamma RII was not affected. In addition, Fc gamma RI+ PMNCs coated in vitro with a bispecific antibody Fab anti-Fc gamma RI x anti-HER-2/neu formed intercellular conjugates with a human HER-2/neu-transrected 3T3 cell line (HER-2/neu-3T3). Interleukin-2 treatment increased the number of Fc gamma RIII low eosinophils, leading to a change in Fc-yRIII distribution among granulocyte cell subsets. In vitro IL-2 treatment of purified PMNCs failed to generate Fc gamma RI expression, suggesting that IL-2 indirectly causes Fc gamma RI expression. During the IL-2 administration, we did not observe significant changes in IFN gamma serum level. In conclusion, our observation may be used to potentiate the antitumor effects of IL-2 in novel immunotherapy regimens, perhaps by redirecting Fc gamma RI+ PMNCs against cancer cells by heteroconjugate antibodies and monitoring the biologic activity of subcutaneous IL-2 in cancer patients. C1 CNR, Inst Tissue Typing & Dialysis, Rome, Italy. Univ Roma Tor Vergata, Dept Surg, Rome, Italy. Univ Roma Tor Vergata, Dept Biopathol, Rome, Italy. Ctr Traumatol Ortoped, Rome, Italy. RP Sconocchia, G (reprint author), NHLBI, Allogene Stem Cell Transplantat Sect, Hematol Branch, NIH, Bldg 10,Room 7C103,9000 Rockville Pike, Bethesda, MD 20892 USA. OI Amadori, Sergio/0000-0002-2356-8681 NR 38 TC 3 Z9 3 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1053-8550 J9 J IMMUNOTHER JI J. Immunother. PD JUL-AUG PY 2001 VL 24 IS 4 BP 374 EP 383 DI 10.1097/00002371-200107000-00013 PG 10 WC Oncology; Immunology; Medicine, Research & Experimental SC Oncology; Immunology; Research & Experimental Medicine GA 469WU UT WOS:000170839700013 PM 11565839 ER PT J AU Thio, CL Thomas, DL Goedert, JJ Vlahov, D Nelson, KE Hilgartner, MW O'Brien, SJ Karacki, P Marti, D Astemborski, J Carrington, M AF Thio, CL Thomas, DL Goedert, JJ Vlahov, D Nelson, KE Hilgartner, MW O'Brien, SJ Karacki, P Marti, D Astemborski, J Carrington, M TI Racial differences in HLA class II associations with hepatitis C virus outcomes SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article; Proceedings Paper CT 51st Annual Meeting of the American-Association-for-the-Study-of-Liver-Diseases CY OCT 27-NOV 01, 2000 CL DALLAS, TX SP Amer Assoc Study Liver Dis ID CHRONIC ACTIVE HEPATITIS; VIRAL CLEARANCE; NATURAL-HISTORY; LONG-TERM; INFECTION; VIREMIA; SUSCEPTIBILITY; HEMOPHILIA; SEQUENCES; GENOTYPE AB A broad, vigorous CD4 T cell response, mediated by class II human leukocyte antigens (HLAs), favors hepatitis C virus (HCV) clearance. HLA-DQB1*0301 has been associated with viral clearance in an ethnically homogeneous cohort. To validate this association and to identify other class II associations in an ethnically varied cohort, molecular class II HLA typing was performed on 200 HCV clearance and 374 matched persistently infected subjects. HLA-DQB1*0301 was weakly associated with viral clearance in combined ethnic groups (odds ratio [OR], 0.72; 95% confidence interval [CI], 0.53-0.97) but was stronger in black subjects. In white subjects, viral clearance was associated with DRB1*0101 (OR, 0.32; 95% CI, 0.17-0.60) and its DQB1*0501 haplotype, whereas viral persistence was associated with DRB1*0301 (OR, 2.36; 95% CI, 1.23-4.52) and its DQB1*0201 haplotype. These results support a role for class II alleles in the immune response to HCV and underscore the importance of studying genetic associations in an ethnically diverse cohort. C1 Johns Hopkins Med Inst, Dept Med, Baltimore, MD 21205 USA. Johns Hopkins Med Inst, Dept Epidemiol, Baltimore, MD 21205 USA. Natl Canc Inst, Viral Epidemiol Branch, Rockville, MD USA. NCI, Lab Genom Divers, Frederick, MD 21701 USA. New York Acad Med, New York, NY USA. New York Presbyterian Hosp, Cornell Med Ctr, Dept Pediat, New York, NY USA. RP Johns Hopkins Univ, Dept Med, 424 N Bond St, Baltimore, MD 21231 USA. EM cthio@jhmi.edu RI Yang, Chen/G-1379-2010 FU NCRR NIH HHS [RR-00071, RR-00059, RR-02558, RR-06020]; NICHD NIH HHS [HD-43200]; NIDA NIH HHS [DA-00441, DA-04334, DA-13324] NR 24 TC 134 Z9 141 U1 2 U2 3 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0022-1899 EI 1537-6613 J9 J INFECT DIS JI J. Infect. Dis. PD JUL 1 PY 2001 VL 184 IS 1 BP 16 EP 21 DI 10.1086/321005 PG 6 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 447DL UT WOS:000169554400003 PM 11398104 ER PT J AU Steel, C Ottesen, EA AF Steel, C Ottesen, EA TI Evolution of immunologic responsiveness of persons living in an area of endemic bancroftian filariasis: A 17-year follow-up SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID PARASITE-SPECIFIC ANERGY; CELLULAR HYPORESPONSIVENESS; BRUGIA-PAHANGI; ANTIGEN; INFECTION; RESPONSES; DIETHYLCARBAMAZINE; MICROFILAREMIA; SUBSET; MALAYI AB On an island in which bancroftian filariasis is endemic, 29 microfilaremic and 16 "endemic normal" (EN) subjects initially studied in 1974-1975 were reevaluated 17 years later. Eleven persons remained microfilaremic, whereas 18 had cleared both microfilaremia and antigenemia. Despite decreased infection on the island, antibody levels remained relatively constant for the subjects with persistent microfilaremia (Mf(+/+)), in contrast to sharp decreases for both EN subjects and subjects with cleared microfilaremia (Mf(+/-)). Although clinically indistinguishable from the EN subjects, the Mf(+/-) group had antibody levels (IgG, IgG4, and IgE) significantly lower than those of the EN subjects. Lymphocyte responses to parasite antigens were marginally greater in Mf(+/-) than in Mf(+/+) subjects, but both groups remained less cell responsive (as measured by proliferation, interleukin-5, interleukin-10, interferon-gamma, and granulocyte- macrophage colony-stimulating factor) than did the EN subjects. These findings suggest that, for microfilaremic persons, complete clearance of infection is not sufficient to restore "normal" immune responsiveness; filarial infection may induce very long-term deficits in the ability to respond to parasite antigens. C1 NIAID, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. WHO, Dept Control Prevent & Eliminat, CH-1211 Geneva, Switzerland. RP Steel, C (reprint author), NIAID, Parasit Dis Lab, NIH, 4 Ctr Dr,Rm 4-126, Bethesda, MD 20892 USA. NR 23 TC 16 Z9 16 U1 0 U2 1 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD JUL 1 PY 2001 VL 184 IS 1 BP 73 EP 79 DI 10.1086/321004 PG 7 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 447DL UT WOS:000169554400011 PM 11398112 ER PT J AU Hengel, RL Jones, BM Kennedy, MS Hubbard, MR McDougal, JS AF Hengel, RL Jones, BM Kennedy, MS Hubbard, MR McDougal, JS TI CD4(+) T cells programmed to traffic to lymph nodes account for increases in numbers of CD4(+) T cells up to 1 year after the initiation of highly active antiretroviral therapy for human immunodeficiency virus type 1 infection SO JOURNAL OF INFECTIOUS DISEASES LA English DT Article ID HOMING RECEPTORS; HIV-1 INFECTION; LYMPHOCYTES; HOMEOSTASIS; DISEASE; SUBSETS AB Cells programmed to traffic through lymph nodes dominate initial increases in total CD4(+) T cell numbers after highly active antiretroviral therapy (HAART) is begun for human immunodeficiency virus type 1 (HIV-1) infection. However, it is unknown whether this dominance continues throughout the first year of treatment. To examine this question, 10 subjects who had a positive response to HAART for 1 year were selected from a cohort of 20 who were receiving this treatment. Flow cytometry, which was used to characterize CD4(+) T cell subsets by immunophenotype, demonstrated that cells programmed to traffic through lymph nodes, irrespective of their memory or naive phenotype, continued to best account for increases in CD4(+) T cells, even 1 year after starting HAART. This suggests that, although this pool is preferentially depleted during HIV-1 infection, HAART allows for reaccumulation of these cells for at least 1 year. Furthermore, it suggests that phenotypic differences based on markers of lymphocyte trafficking may be more relevant for understanding HIV-1 pathogenesis than are naive and memory markers alone. C1 Emory Univ, Sch Med, Dept Med, Div Infect Dis, Atlanta, GA USA. Ctr Dis Control & Prevent, Natl Ctr Infect Dis, Div AIDS STD & TB Lab Res, Immunol Branch, Atlanta, GA USA. RP Hengel, RL (reprint author), NIAID, LIR, NIH, Bldg 10,11B05,MSC 1880,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 15 TC 10 Z9 10 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 0022-1899 J9 J INFECT DIS JI J. Infect. Dis. PD JUL 1 PY 2001 VL 184 IS 1 BP 93 EP 97 DI 10.1086/320997 PG 5 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 447DL UT WOS:000169554400014 PM 11398115 ER PT J AU Mudd, SH Cerone, R Schiaffino, MC Fantasia, AR Minniti, G Caruso, U Lorini, R Watkins, D Matiaszuk, N Rosenblatt, DS Schwahn, B Rozen, R LeGros, L Kotb, M Capdevila, A Luka, Z Finkelstein, JD Tangerman, A Stabler, SP Allen, RH Wagner, C AF Mudd, SH Cerone, R Schiaffino, MC Fantasia, AR Minniti, G Caruso, U Lorini, R Watkins, D Matiaszuk, N Rosenblatt, DS Schwahn, B Rozen, R LeGros, L Kotb, M Capdevila, A Luka, Z Finkelstein, JD Tangerman, A Stabler, SP Allen, RH Wagner, C TI Glycine N-methyltransferase deficiency: A novel inborn error causing persistent isolated hypermethioninaemia SO JOURNAL OF INHERITED METABOLIC DISEASE LA English DT Article ID METHIONINE ADENOSYLTRANSFERASE DEFICIENCY; METHYLENETETRAHYDROFOLATE REDUCTASE; S-ADENOSYLMETHIONINE; RAT-LIVER; CREATINE DEFICIENCY; FOLATE-DEFICIENCY; METABOLISM; PLASMA; DISEASE; CHOLINE AB This paper reports clinical and metabolic studies of two Italian siblings with a novel form of persistent isolated hypermethioninaemia, i.e. abnormally elevated plasma methionine that lasted beyond the first months of life and is not due to cystathionine beta -synthase deficiency, tyrosinaemia I or liver disease. Abnormal elevations of their plasma S-adenosylmethionine (AdoMet) concentrations proved they do not have deficient activity of methionine adenosyltransferase I/III. A variety of studies provided evidence that the elevations of methionine and AdoMet are not caused by defects in the methionine transamination pathway, deficient activity of methionine adenosyltransferase II, a mutation in methylenetetrahydrofolate reductase rendering this activity resistant to inhibition by AdoMet, or deficient activity of guanidinoacetate methyltransferase. Plasma sarcosine (N-methylglycine) is elevated, together with elevated plasma AdoMet in normal subjects following oral methionine loads and in association with increased plasma levels of both methionine and AdoMet in cystathionine beta -synthase-deficient individuals. However, plasma sarcosine is not elevated in these siblings. The latter result provides evidence they are deficient in activity of glycine N-methyltransferase (GNMT). The only clinical abnormalities in these siblings are mild hepatomegaly and chronic elevation of serum transaminases not attributable to conventional causes of liver disease. A possible causative connection between GNMT deficiency and these hepatitis-like manifestations is discussed. Further studies are required to evaluate whether dietary methionine restriction will be useful in this situation. C1 NIMH, DIRP, LMB, Bethesda, MD 20892 USA. Univ Genoa, Dept Pediat, G Gaslini Inst, Genoa, Italy. McGill Univ, Ctr Hlth, Dept Med, Div Med Genet, Montreal, PQ, Canada. McGill Univ, Ctr Hlth, Dept Pediat, Montreal, PQ, Canada. McGill Univ, Ctr Hlth, Dept Human Genet, Montreal, PQ, Canada. McGill Univ, Ctr Hlth, Dept Biol, Montreal, PQ, Canada. Vet Adm Med Ctr, Memphis, TN 38104 USA. Univ Tennessee, Dept Surg, Memphis, TN USA. Univ Tennessee, Dept Microbiol & Immunol, Memphis, TN USA. Vanderbilt Univ, Dept Biochem, Nashville, TN 37232 USA. Vanderbilt Univ, Clin Nutr Res Unit, Nashville, TN 37232 USA. Vet Adm Med Ctr, Washington, DC 20422 USA. George Washington Univ, Washington, DC USA. Univ Nijmegen Hosp, Div Gastroenterol & Hepatol, NL-6500 HB Nijmegen, Netherlands. Univ Colorado, Hlth Sci Ctr, Dept Med, Denver, CO 80262 USA. Dept Vet Affairs Med Ctr, Nashville, TN 37212 USA. RP Mudd, SH (reprint author), NIMH, DIRP, LMB, Bldg 36,Room 1B-08,36 Convent Dr MSC 4034, Bethesda, MD 20892 USA. EM shm@codon.nih.gov FU NIA NIH HHS [AG-09834]; NIDDK NIH HHS [DK15289, DK54859] NR 62 TC 84 Z9 84 U1 0 U2 5 PU SPRINGER PI DORDRECHT PA VAN GODEWIJCKSTRAAT 30, 3311 GZ DORDRECHT, NETHERLANDS SN 0141-8955 J9 J INHERIT METAB DIS JI J. Inherit. Metab. Dis. PD JUL PY 2001 VL 24 IS 4 BP 448 EP 464 DI 10.1023/A:1010577512912 PG 17 WC Endocrinology & Metabolism; Genetics & Heredity; Medicine, Research & Experimental SC Endocrinology & Metabolism; Genetics & Heredity; Research & Experimental Medicine GA 460BG UT WOS:000170285500003 PM 11596649 ER PT J AU Weinberg, NZ AF Weinberg, NZ TI Risk factors for adolescent substance abuse SO JOURNAL OF LEARNING DISABILITIES LA English DT Article ID POPULATION-BASED SAMPLE; USE DISORDERS; FOLLOW-UP; DRUG-ABUSE; DEVELOPMENTAL PSYCHOPATHOLOGY; AGGRESSIVE-BEHAVIOR; ALCOHOL DEPENDENCE; FEMALE TWINS; BOYS; VULNERABILITY AB Reviews research on risk factors for adolescent substance use disorders (SUD) and discusses possible relationships between SUDs and learning disabilities (LD). Individual level factors (genetic, biologic, other familial, and psychiatric) emerge as very important in the risk equation, as well as the interaction between individual risk and environmental conditions. Commonalities between SUD risk and LD include prenatal substance exposure, family history of SUD, conduct disorder, social skills deficits, and academic failure; however, further research is needed to establish whether individuals with LD face a specific risk for SUDs, and if so, what the nature of that risk might be. C1 NIDA, Div Epidemiol Serv & Prevent Res, NIH, Bethesda, MD 20892 USA. RP Weinberg, NZ (reprint author), NIDA, Div Epidemiol Serv & Prevent Res, NIH, 6001 Execut Blvd,Rm 5153,MSC 9589, Bethesda, MD 20892 USA. NR 64 TC 16 Z9 17 U1 2 U2 8 PU PRO-ED INC PI AUSTIN PA 8700 SHOAL CREEK BLVD, AUSTIN, TX 78757-6897 USA SN 0022-2194 J9 J LEARN DISABIL-US JI J. Learn. Disabil. PD JUL-AUG PY 2001 VL 34 IS 4 BP 343 EP 351 DI 10.1177/002221940103400409 PG 9 WC Education, Special; Rehabilitation SC Education & Educational Research; Rehabilitation GA 452CQ UT WOS:000169839900009 PM 15503578 ER PT J AU Smyth, MJ Kelly, JM Sutton, VR Davis, JE Browne, KA Sayers, TJ Trapani, JA AF Smyth, MJ Kelly, JM Sutton, VR Davis, JE Browne, KA Sayers, TJ Trapani, JA TI Unlocking the secrets of cytotoxic granule proteins SO JOURNAL OF LEUKOCYTE BIOLOGY LA English DT Review DE apoptosis; lymphocyte; granzyme; perforin; granulysin ID NATURAL-KILLER-CELLS; CYTOLYTIC-T-LYMPHOCYTES; PERFORIN-DEFICIENT MICE; DIPEPTIDYL PEPTIDASE-I; MEDIATED CYTO-TOXICITY; TUMOR-NECROSIS-FACTOR; PORE-FORMING PROTEIN; VERSUS-HOST DISEASE; HUMAN GRANZYME-A; SERINE PROTEASES AB Cytotoxic lymphocytes largely comprise CD8(+) cytotoxic T cells and natural killer cells and form the major defense of higher organisms against virus-infected and transformed cells, A key function of cytotoxic lymphocytes is to detect and eliminate potentially harmful cells by inducing them to undergo apoptosis, This is achieved through two principal pathways, both of which require direct but transient contact between the killer cell and its target. The first, involving ligation of TNF receptor-like molecules such as Fas/CD95 by their cognate ligands, results in mobilization of conventional, programmed cell-death pathways centered on activation of pro-apoptotic caspases, This review concentrates on the second pathway, in which the toxic contents of secretory vesicles of the cytotoxic lymphocyte are secreted toward the target cell, and some toxins penetrate into the target cell cytoplasm and nucleus. In addition to invoking a powerful stimulus to caspase activation, this "granule-exocytosis mechanism" provides a variety of additional strategies for overcoming inhibitors of the caspase cascade that may be elaborated by viruses. The key molecular players in this process are the pore-forming protein perforin and a family of granule-bound serine proteases or granzymes, The molecular functions of perforin and granzymes are under intense investigation in many laboratories including our own, and recent advances will be discussed. In addition, this review discusses the evidence pointing to the importance of perforin and granzyme function in pathophysiological situations as diverse as infection with intracellular pathogens, graft versus host disease, susceptibility to transplantable and spontaneous malignancies, lymphoid homeostasis, and the tendency to auto-immune diseases. C1 Peter MacCallum Canc Inst, Can Immunol Div, Trescowthick Labs, Melbourne, Vic 8006, Australia. NCI, Expt Immunol Lab, FDR, DC,NIH, Frederick, MD 21701 USA. RP Smyth, MJ (reprint author), Peter MacCallum Canc Inst, Can Immunol Div, Trescowthick Labs, Locked Bag 1,A Beckett St, Melbourne, Vic 8006, Australia. RI Sayers, Thomas/G-4859-2015; Smyth, Mark/H-8709-2014 OI Smyth, Mark/0000-0001-7098-7240 FU NCI NIH HHS [N01-CO-56000] NR 133 TC 116 Z9 121 U1 1 U2 8 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 JUL PY 2001 VL 70 IS 1 BP 18 EP 29 PG 12 WC Cell Biology; Hematology; Immunology SC Cell Biology; Hematology; Immunology GA 451JU UT WOS:000169799600003 PM 11435481 ER PT J AU Hu, JY Le, YY Gong, WH Dunlop, NM Gao, JL Murphy, PM Wang, JM AF Hu, JY Le, YY Gong, WH Dunlop, NM Gao, JL Murphy, PM Wang, JM TI Synthetic peptide MMK-1 is a highly specific chemotactic agonist for leukocyte FPRL1 SO JOURNAL OF LEUKOCYTE BIOLOGY LA English DT Article DE phagocyte; chemotaxis; Ca(2+) mobilization; cytokines ID SERUM AMYLOID-A; PROTEIN-COUPLED RECEPTOR; PHOSPHOINOSITIDE HYDROLYSIS; SUPEROXIDE GENERATION; HUMAN NEUTROPHILS; HUMAN PHAGOCYTES; HUMAN MONOCYTES; GENE-CLUSTER; IDENTIFICATION; ACTIVATION AB Human phagocytic leukocytes express the seven-transmembrane G-protein-coupled receptors formyl peptide receptor (FPR) and FPR-like 1 (FPRL1). MMK-1, a synthetic peptide derived from a random peptide library, is reported to induce calcium mobilization specifically in human FPRL1 gene-transfected cells. However, its actions on human phagocytic leukocytes remain poorly defined. We found that NMK-1 is a potent chemotactic and calcium-mobilizing agonist for human monocytes, neutrophils, and FPRL1-transfected human embryonic kidney (HEK) 293 cells but is inactive in cells transfected with FPR. MMK-1 also activated HEK 293 cells transfected with FPR2, a mouse counterpart of human FPRL1. Furthermore, MMK-1 increased pertussis toxin-sensitive production of inflammatory cytokines in human monocytes. MMK-1 signaling in human phagocytes was completely desensitized by a well-defined FPRL1 agonist, suggesting that FPRL1 is likely a receptor that mediates the action of MMK-1 in primary cells. Since MMK-1 is one of the most potent FPRL1-specific agonists identified so far, it can serve as a modulator of the host defense and a useful agent for further studying the signaling and function of FPRL1. C1 NCI, Frederick Canc Res & Dev Ctr, LMI, DBS, Frederick, MD 21702 USA. SAIC, Intramural Res Support Program, Frederick, MD USA. NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. Cent S Univ, Canc Res Inst, Changsha, Peoples R China. RP Wang, JM (reprint author), NCI, Frederick Canc Res & Dev Ctr, LMI, DBS, Bldg 560,Room 31-40, Frederick, MD 21702 USA. EM wangji@mail.ncifcrf.gov FU NCI NIH HHS [N01-CO-56000] NR 30 TC 67 Z9 67 U1 0 U2 0 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 JUL PY 2001 VL 70 IS 1 BP 155 EP 161 PG 7 WC Cell Biology; Hematology; Immunology SC Cell Biology; Hematology; Immunology GA 451JU UT WOS:000169799600021 PM 11435499 ER PT J AU Dean, M Hamon, Y Chimini, G AF Dean, M Hamon, Y Chimini, G TI The human ATP-binding cassette (ABC) transporter superfamily SO JOURNAL OF LIPID RESEARCH LA English DT Review DE membrane transporters; evolution; lipid; genetic diseases; cholesterol ID FAMILIAL INTRAHEPATIC CHOLESTASIS; RESISTANCE-ASSOCIATED PROTEIN; DROSOPHILA-WHITE GENE; MOUSE P-GLYCOPROTEIN; CYSTIC-FIBROSIS GENE; MULTIDRUG-RESISTANCE; PLASMA-MEMBRANE; RIM PROTEIN; BACTERIAL TRANSPORT; ELECTRON-MICROSCOPY AB The transport of specific molecules across lipid membranes is an essential function of all living organisms and a large number of specific transporters have evolved to carry out this function. The largest transporter gene family is the ATP-binding cassette (ABC) transporter superfamily, These proteins translocate a wide variety of substrates including sugars, amino acids, metal ions, peptides, and proteins, and a large number of hydrophobic compounds and metabolites across extra- and intracellular membranes. ABC genes are essential for many processes in the cell, and mutations in these genes cause or contribute to several human genetic disorders including cystic fibrosis, neurological disease, renal degeneration, cholesterol and bile transport defects, anemia, and drug response. Characterization of eukaryotic genomes has allowed the complete identification of all the ABC genes in the yeast Saccharomyces cerevisiae, Drosophila, and C. elegans genomes, To date, there are 48 characterized human ABC genes. The genes can be divided into seven distinct subfamilies, based on organization of domains and amino acid homology. Many ABC genes play a role in the maintenance of the lipid bilayer and in the transport of fatty acids and sterols within the body. jlr Here, we review the current knowledge of the human ABC genes, their role in inherited disease, and understanding of the topology of these genes within the membrane. C1 NCI, Human Genet Sect, Lab Genom Divers, Frederick, MD 21702 USA. CNRS Marseille Luminy, Ctr Immunol, INSERM, F-13288 Marseille 09, France. RP Dean, M (reprint author), NCI, Human Genet Sect, Lab Genom Divers, Bldg 560,Rm 21-18, Frederick, MD 21702 USA. EM dean@ncifcrf.gov RI Dean, Michael/G-8172-2012; HAMON, Yannick/N-5372-2016 OI Dean, Michael/0000-0003-2234-0631; HAMON, Yannick/0000-0002-7636-2310 NR 106 TC 627 Z9 680 U1 4 U2 49 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0022-2275 J9 J LIPID RES JI J. Lipid Res. PD JUL PY 2001 VL 42 IS 7 BP 1007 EP 1017 PG 11 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 451CV UT WOS:000169784300001 PM 11441126 ER PT J AU Hung, CL Doniger, J Palini, A Snyder, SW Radonovich, MF Brady, JN Pantazis, P Sadaie, MR AF Hung, CL Doniger, J Palini, A Snyder, SW Radonovich, MF Brady, JN Pantazis, P Sadaie, MR TI 9-Nitrocamptothecin inhibits HIV-1 replication in human peripheral blood lymphocytes: A potential alternative for HIV-Infection/AIDS therapy SO JOURNAL OF MEDICAL VIROLOGY LA English DT Article DE antiviral drugs; 9-Nitrocamptothecin; human peripheral blood lymphocytes; HIV/AIDS ID HUMAN-IMMUNODEFICIENCY-VIRUS; INSOLUBLE CAMPTOTHECIN ANALOGS; TOPOISOMERASE-I; CELLS; TYPE-1; DRUGS; 9-NITRO-CAMPTOTHECIN; PHARMACOKINETICS; CYTOTOXICITY; DERIVATIVES AB The ability of the anti-cancer drug, 9-Nitro-camptothecin (9NC), to inhibit replication of HIV-1 in clinically relevant primary lymphocytic cells was studied. Primary peripheral blood lymphocytes (PBLs) from a non-infected donor were freshly infected with HIV-1 and treated with 9NC by using three different treatment schedules. Cells were monitored for cytotoxicity by the XTT metabolic cell proliferation assay and a sensitive flow cytometric assay that was capable of measuring cell cycle changes and apoptosis. 9NC inhibited replication of HIV-1 in PBLs by greater than 95% in a dose-dependent manner as measured by the level of extracellular HIV-1 p24 release. Similar results were observed, whether 9NC was applied in a single, double, or triple dose regimen. Minimal cytotoxicity was observed for both non-infected and infected PBLs, as determined by the XTT assay. Moreover, 9NC induced apoptosis within 24 hours of drug treatment in freshly infected, but not non-infected, PBLs. The data showed that 9NC reduced replication of HIV-1 in primary human lymphocytes; thus, it indicates the potential clinical utility of this drug as an alternative or adjunct therapy for HIV-infection/AIDS. J. Med. Virol. 64:238-244, 2001. (C) 2001 Wiley-Liss, Inc. C1 NovoMed Pharmaceut Inc, Germantown, MD 20875 USA. Adv Biosci Labs, Kensington, NSW, Australia. FAST Syst Inc, Gaithersburg, MD USA. Univ Washington, Sch Med, Retrovirus Lab, Seattle, WA 98195 USA. NCI, Lab Receptor Biol & Gene Express, NIH, Bethesda, MD 20892 USA. Brown Univ, Div Biol & Med, Dept Biochem Mol Biol & Cell Biol, Providence, RI 02912 USA. RP Sadaie, MR (reprint author), NovoMed Pharmaceut Inc, POB 900, Germantown, MD 20875 USA. NR 36 TC 10 Z9 22 U1 2 U2 3 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0146-6615 J9 J MED VIROL JI J. Med. Virol. PD JUL PY 2001 VL 64 IS 3 BP 238 EP 244 DI 10.1002/jmv.1042 PG 7 WC Virology SC Virology GA 437ZQ UT WOS:000169029600006 PM 11424110 ER PT J AU Kaiser, L Fritz, RS Straus, SE Gubareva, L Hayden, FG AF Kaiser, L Fritz, RS Straus, SE Gubareva, L Hayden, FG TI Symptom pathogenesis during acute influenza: Interleukin-6 and other cytokine responses SO JOURNAL OF MEDICAL VIROLOGY LA English DT Article DE influenza; interleukin-6; tumor necrosis factor-alpha; interferon-alpha; interferon-gamma; interleukin-10; interleukin-8; influenza virus A; oseltamivir; cytokines ID NEURAMINIDASE INHIBITOR OSELTAMIVIR; A VIRUS-INFECTION; VIRAL-INFECTIONS; GAMMA-PRODUCTION; LYMPHOCYTES; IMMUNITY; CHILDREN; DEFENSE; TRIAL; BLOOD AB In experimental human influenza infection initiated by nasal inoculation, the magnitude of viral replication, fever, and symptoms correlate with nasopharyngeal lavage fluid levels of various cytokines. Our aim was to assess these relationships in patients with naturally occurring acute influenza. Patients with culture-positive influenza illness of less than 36 hr of duration were studied. Nasopharyngeal washing were collected at enrollment and on Day 2, 4, 6 and 8 for quantitative virus isolation and IL-6, TNF-alpha, INF-alpha, INF-gamma and IL-10 determinations. Blood samples collected at entry and on Day 2 and 6 were processed to assess plasma cytokines and circulating influenza RNA. Patients received either oseltamivir or placebo for 5 days. We assessed the correlation between nasopharyngeal lavage fluid or blood levels of cytokines before treatment and viral titers, symptom severity and fever. Sixteen adult subjects (median age of 22 years) were studied, in this small group of patients no significant differences between placebo and oseltamivir patients were found in viral replication or measures of cytokines. Thus the data for all 16 subjects were pooled for analysis. At entry, influenza A viruses were cultured from nasopharyngeal washes at a median titer of 4.8 log(10)TCID(50)/ml of wash. Viral titers correlated positively with symptom score (P=0.006) and temperature values (P <0.001). Viral titers, fever and symptoms were highest at enrollment and fell in parallel during the subsequent days. RT-PCR assays failed to detect influenza RNA in the white blood eel Is from any patient. We observed a significant release, in both nasopharyngeal lavage fluid and in plasma, of IL-6, TNF-alpha, INF-alpha, INF-gamma and IL-10. At entry high IL-6 levels were detected in the nasopharyngeal lavage fluid (median 10.3 pg/ml) and plasma (median 5.1 pg/ml) of all patients. We found a positive correlation between plasma IL-6 levels and both symptom scores and temperature values (P <0.05), as well as a positive correlation between nasopharyngeal lavage fluid levels of IL-6 and TNF-alpha and temperature (P <0.05). We did not find significant associations between symptoms, fever and levels of INF-alpha, INF-gamma or IL-10. The magnitude of early decrease in viral titers correlated with initial levels of INF-gamma in nasopharyngeal ravage fluid (P <0.05). Significant production of IL-6, TNF-alpha, INF-alpha ,INF-gamma and IL-10 occurs in response to community acquired influenza A illness. As in experimental influenza, symptoms and fever in natural acute influenza correlate with the release of IL-6. J. Med. Virol. 64:262-268, 2001. (C) 2001 Wiley-Liss, Inc. C1 Univ Virginia, Hlth Sci Ctr, Sch Med, Dept Internal Med,Div epidemiol & Virol, Charlottesville, VA 22908 USA. NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, Frederick, MD 21702 USA. NIAID, Clin Invest Lab, Bethesda, MD 20892 USA. RP Hayden, FG (reprint author), Univ Virginia, Hlth Sci Ctr, Sch Med, Dept Internal Med,Div epidemiol & Virol, Box 473, Charlottesville, VA 22908 USA. NR 21 TC 177 Z9 185 U1 0 U2 4 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0146-6615 J9 J MED VIROL JI J. Med. Virol. PD JUL PY 2001 VL 64 IS 3 BP 262 EP 268 DI 10.1002/jmv.1045 PG 7 WC Virology SC Virology GA 437ZQ UT WOS:000169029600009 PM 11424113 ER PT J AU McDermott, J Beck, D Buffington, ST Annas, J Supratikto, G Prenggono, D Ekonomi, MFS Achadi, E AF McDermott, J Beck, D Buffington, ST Annas, J Supratikto, G Prenggono, D Ekonomi, MFS Achadi, E TI Two models of in-service training to improve midwifery skills: How well do they work? SO JOURNAL OF MIDWIFERY & WOMENS HEALTH LA English DT Article AB This program evaluation compared the knowledge, confidence, and skills of Indonesian village midwives who attended an intensive in-service training with midwives who received an internship program and midwives who attended no program. The five key skills compared were prevention of infection, use of the partograph, manual removal of placenta, bimanual uterine compression, and neonatal resuscitation. Midwives from the intensive in-service that combined competency-based skill training with peer review and continuing education scored higher on the knowledge test and demonstration of the five key skills and reported managing complications better than midwives who attended no training program. Midwives from the internship program scored intermediate between the intensively trained and the untrained midwives. Overall, skill scores were 71% for midwives in the intensive program, 62% for the interns, and 51% for midwives with no in-service training. Village midwives from the intensive program scored significantly higher in the practical demonstration of manual removal of placenta, bimanual compression, and neonatal resuscitation than the interns, but the scores on infection prevention and use of the partograph were not different between the two groups. Differences in the Volume of training opportunities between the two programs could be responsible for the different outcomes. (C) 2001 by the American College of Nurse-Midwives. C1 NIH, Div Int Training & Res, Fogarty Int Ctr, Bethesda, MD USA. RP McDermott, J (reprint author), 4819 Western Ave NW, Washington, DC 20016 USA. NR 6 TC 19 Z9 19 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 1526-9523 J9 J MIDWIFERY WOM HEAL JI J. Midwifery Women Health PD JUL-AUG PY 2001 VL 46 IS 4 BP 217 EP 225 DI 10.1016/S1526-9523(01)00137-4 PG 9 WC Nursing SC Nursing GA 481RR UT WOS:000171534100004 PM 11603633 ER PT J AU Rosenberg, HF Zhang, JZ Liao, YD Dyer, KD AF Rosenberg, HF Zhang, JZ Liao, YD Dyer, KD TI Rapid diversification of RNase a superfamily ribonucleases from the bullfrog, Rana catesbeiana SO JOURNAL OF MOLECULAR EVOLUTION LA English DT Article DE ribonucleases; molecular evolution; amphibians ID AMINO-ACID-SEQUENCE; NONSYNONYMOUS NUCLEOTIDE SUBSTITUTIONS; CYTOTOXIC RIBONUCLEASE; A SUPERFAMILY; EVOLUTION; PURIFICATION; PROTEIN; CLONING; LECTIN; RODENT AB We present sequences of five novel RNase A superfamily ribonuclease genes of the bullfrog, Rana catesbeiana. All five genes encode ribonucleases that are similar to Onconase, a cytotoxic ribonuclease isolated from oocytes of R, pipiens. With amino acid sequence data from 14 ribonucleases from three Rana species (R, catesbeiana, R. japonica, and R. pipiens), we have constructed bootstrap-supported phylogenetic trees that reorganize these ribonucleases into five distinct lineages-the pancreatic ribonucleases (RNases 1), the eosinophil-associated ribonucleases (RNases 2, 3, and 6), the ribonucleases 4, the angiogenins (RNases 5) and the Rana ribonucleases-with the Rana ribonucleases no more closely related to the angiogenins than they are to any of the other ribonuclease lineages shown. Further phylogenetic analysis suggests the division of the Rana ribonucleases into two subclusters (A and B), with positive (Darwinian) selection (d(N)/d(S) > 1.0) and an elevated rate of radical nonsynonymous substitution (d(R)) contributing to the rapid diversification of ribonucleases within each cluster. This pattern of evolution-rapid diversification via positive selection among sequences of a multigene cluster-bears striking resemblance to what we have described for the eosinophil-associated ribonuclease genes of the rodent Mus musculus, a finding that may have implications with respect the physiologic function of this unique family of proteins. C1 NIAID, LHD, NIH, Bethesda, MD 20892 USA. Acad Sinica, Inst Biomed Sci, Taipei 115, Taiwan. RP Rosenberg, HF (reprint author), NIAID, LHD, NIH, Bldg 10,Room 11N104,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 24 TC 21 Z9 24 U1 0 U2 2 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0022-2844 J9 J MOL EVOL JI J. Mol. Evol. PD JUL PY 2001 VL 53 IS 1 BP 31 EP 38 PG 8 WC Biochemistry & Molecular Biology; Evolutionary Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Evolutionary Biology; Genetics & Heredity GA 446ZN UT WOS:000169544200004 PM 11683320 ER PT J AU Ellsworth, DL Bielak, LF Turner, ST Sheedy, PF Boerwinkle, E Peyser, PA AF Ellsworth, DL Bielak, LF Turner, ST Sheedy, PF Boerwinkle, E Peyser, PA TI Gender- and age-dependent relationships between the E-selectin S128R polymorphism and coronary artery calcification SO JOURNAL OF MOLECULAR MEDICINE-JMM LA English DT Article DE atherosclerosis (coronary); calcium; cellular adhesion molecules; computed tomography; genetics; heart disease ID INTERCELLULAR-ADHESION MOLECULE-1; BEAM COMPUTED-TOMOGRAPHY; CELL-ADHESION; POSTMENOPAUSAL WOMEN; HEART-DISEASE; RISK-FACTORS; MYOCARDIAL-INFARCTION; ATHEROSCLEROSIS; LIPOPROTEINS; PERSPECTIVE AB Development and progression of atherosclerosis involves recruitment and binding of circulating leukocytes to areas of inflammation within the vascular endothelium mediated by a diverse array of cellular adhesion molecules. A polymorphism in the endothelial-leukocyte adhesion molecule 1 (E-selectin) gene has been implicated in early-onset, angiographically defined, severe atherosclerotic disease because it profoundly affects ligand recognition and binding specificity, resulting in a significant increase in cellular adhesion. Relationships between the E-selectin S128R polymorphism and coronary artery calcification (CAC), a marker of atherosclerosis detected with noninvasive electron beam computed tomography, were examined in 294 asymptomatic women aged 40-88 years and 314 asymptomatic men aged 30-80 years from the Epidemiology of Coronary Artery Calcification Study. The E-selectin polymorphism was not associated with presence of CAC in men of any age or in women over age 50. In women 50 years of age or younger the E-selectin polymorphism was significantly associated with presence of CAC after adjustment for age, body mass index, systolic blood pressure, ratio of total cholesterol to high-density lipoprotein cholesterol, and smoking. The significant association between E-selectin and CAC in women 50 years of age or younger may suggest that the 128R allele is a risk factor for coronary atherosclerosis in younger asymptomatic women, who typically have lower levels of traditional risk factors and reduced adhesion molecule expression due to the presence of higher levels of endogenous hormones. C1 Univ Michigan, Dept Epidemiol, Ann Arbor, MI 48109 USA. NHLBI, Div Epidemiol & Clin Applicat, NIH, Bethesda, MD 20892 USA. Mayo Clin, Div Hypertens, Rochester, MN USA. Mayo Clin, Dept Internal Med, Rochester, MN USA. Mayo Clin, Dept Diagnost Radiol, Rochester, MN USA. Univ Texas, Hlth Sci Ctr, Ctr Human Genet, Houston, TX USA. RP Peyser, PA (reprint author), Univ Michigan, Dept Epidemiol, 109 Observ, Ann Arbor, MI 48109 USA. FU NHLBI NIH HHS [HL46292] NR 53 TC 54 Z9 67 U1 0 U2 1 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0946-2716 J9 J MOL MED-JMM JI J. Mol. Med. PD JUL PY 2001 VL 79 IS 7 BP 390 EP 398 DI 10.1007/s001090100235 PG 9 WC Genetics & Heredity; Medicine, Research & Experimental SC Genetics & Heredity; Research & Experimental Medicine GA 457DT UT WOS:000170122400008 PM 11466561 ER PT J AU Peterkofsky, A Wang, GS Garrett, DS Lee, BR Seok, YJ Clore, GM AF Peterkofsky, A Wang, GS Garrett, DS Lee, BR Seok, YJ Clore, GM TI Three-dimensional structures of protein-protein complexes in the E-coli PTS SO JOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY LA English DT Article ID SUGAR PHOSPHOTRANSFERASE SYSTEM; N-TERMINAL DOMAIN; CONTAINING PHOSPHOCARRIER PROTEIN; PHOSPHORYL TRANSFER COMPLEX; ESCHERICHIA-COLI; ENZYME-I; HPR; TRANSPORT; PHOSPHOENOLPYRUVATE; BINDING AB The bacterial phosphoenolpyruvate:sugar phosphotransferase system (PTS) includes a collection of proteins that accomplish phosphoryl transfer from phosphoenolpyruvate (PEP) to a sugar in the course of transport. The soluble proteins of the glucose transport pathway also function as regulators of diverse systems. The mechanism of interaction of the phosphoryl carrier proteins with each other as well as with their regulation targets has been amenable to study by nuclear magnetic resonance (NMR) spectroscopy. The three-dimensional solution structures of the complexes between the N-terminal domain of enzyme I and HPr and between HPr and enzyme IIA(Glc) have been elucidated. An analysis of the binding interfaces of HPr with enzyme I, IIA(Glc) and glycogen phosphorylase revealed that a common surface on HPr is involved in all these interactions. Similarly, a common surface on IIA(Glc) interacts with HPr, IIBGlc and glycerol kinase. Thus, there is a common motif for the protein-protein interactions characteristic of the PTS. C1 NHLBI, Lab Biochem Genet, NIH, Bethesda, MD 20892 USA. NIDDK, Chem Phys Lab, Bethesda, MD 20892 USA. Seoul Natl Univ, Sch Biol Sci, Seoul 151742, South Korea. RP Peterkofsky, A (reprint author), NHLBI, Lab Biochem Genet, NIH, Bldg 36,Rm 4C-11, Bethesda, MD 20892 USA. RI Clore, G. Marius/A-3511-2008 OI Clore, G. Marius/0000-0003-3809-1027 NR 15 TC 27 Z9 27 U1 0 U2 2 PU HORIZON SCIENTIFIC PRESS PI WYMONDHAM PA PO BOX 1, NORFOLK, WYMONDHAM NR18 0JA, ENGLAND SN 1464-1801 J9 J MOL MICROB BIOTECH JI J. Mol. Microbiol. Biotechnol. PD JUL PY 2001 VL 3 IS 3 BP 347 EP 354 PG 8 WC Biotechnology & Applied Microbiology; Microbiology SC Biotechnology & Applied Microbiology; Microbiology GA 429AU UT WOS:000168494500003 PM 11361064 ER PT J AU Seok, YJ Koo, BM Sondej, M Peterkofsky, A AF Seok, YJ Koo, BM Sondej, M Peterkofsky, A TI Regulation of E-coli glycogen phosphorylase activity by HPr SO JOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY LA English DT Article ID SUGAR PHOSPHOTRANSFERASE SYSTEM; CONTAINING PHOSPHOCARRIER PROTEIN; GRAM-POSITIVE BACTERIA; N-TERMINAL DOMAIN; ESCHERICHIA-COLI; ENZYME-I; LACTOSE PERMEASE; SALMONELLA-TYPHIMURIUM; ALLOSTERIC REGULATION; RESOLUTION STRUCTURE AB Bacteria sense continuous changes in their environment and adapt metabolically to effectively compete with other organisms for limiting nutrients. One system which plays an important part in this adaptation response is the phosphoenolpyruvate:sugar phosphotransferase system (PTS). Many proteins interact with and are regulated by PTS components in bacteria. Here we review the interaction with and allosteric regulation of Escherichia coli glycogen phosphorylase (GP) activity by the histidine phosphocarrier protein HPr, which acts as part of a phosphoryl shuttle between enzyme I and sugar-specific proteins of the PTS. HPr mediates crosstalk between PTS sugar uptake and glycogen breakdown. The evolution of the allosteric regulation of E. coli GP by HPr is compared to that of other phosphorylases. C1 Seoul Natl Univ, Sch Biol Sci, Seoul 151742, South Korea. NHLBI, Lab Biochem Genet, NIH, Bethesda, MD 20892 USA. RP Seok, YJ (reprint author), Seoul Natl Univ, Sch Biol Sci, Seoul 151742, South Korea. NR 52 TC 24 Z9 24 U1 1 U2 4 PU HORIZON SCIENTIFIC PRESS PI WYMONDHAM PA PO BOX 1, NORFOLK, WYMONDHAM NR18 0JA, ENGLAND SN 1464-1801 J9 J MOL MICROB BIOTECH JI J. Mol. Microbiol. Biotechnol. PD JUL PY 2001 VL 3 IS 3 BP 385 EP 393 PG 9 WC Biotechnology & Applied Microbiology; Microbiology SC Biotechnology & Applied Microbiology; Microbiology GA 429AU UT WOS:000168494500008 PM 11361069 ER PT J AU Stiles, KP Hill, C LeBrun, C Reinmuth, B Yuan, CM Abbott, KC AF Stiles, KP Hill, C LeBrun, C Reinmuth, B Yuan, CM Abbott, KC TI The impact of bleeding times on major complication rates after percutaneous real-time ultrasound-guided renal biopsies SO JOURNAL OF NEPHROLOGY LA English DT Article DE renal biopsy; bleeding time; risk; real-time ultrasound guidance; spring loaded biopsy needle; complications ID DISEASES; PATIENT; KIDNEY AB Background. Previous studies have shown that bleeding times have positive predictive values of only 5% for perioperative bleeding in unselected populations. Nevertheless, performing bleeding times prior to all renal biopsies is common in nephrology practice. Methods. We report complications of 112 renal biopsies done at Walter Reed Army Medical Center (WRAMC) from 1996-99 performed without preceding bleeding times. Renal biopsies were done only on normotensive (<140/90) patients who had not recently been taking aspirin or non-steroidal anti-inflammatory agents, under real-time ultrasound guidance with automated 16 g (WRAMC) spring-loaded guns. High-risk patients (with serum creatinine 3 mg/dl or creatinine clearance less than or equal to 30 cc/min by Cockroft-Gault formula, N=18, 16%) at WRAMC were treated with pre-renal biopsy estrogens or DDAVP. Factors were tested for their association with complications after renal biopsy using Chi Square testing for categorical variables and student's t-test for continuous variables. A stepwise logistic regression model was used to test for independent significance of factors. Results. There were two cases each of gross hematuria and inadequate tissue (1.8% each). There were no transfusions or deaths. In univariate analysis, male gender and lower serum creatinine level at time of biopsy were significantly associated with increased risk of complications after biopsy. However, these factors were not significant in logistic regression analysis. Conclusion. This study suggests that the use of bleeding times does not significantly alter the major complication rates associated with percutaneous real-time ultrasound guided renal biopsy. C1 Walter Reed Army Med Ctr, Serv Nephrol, Washington, DC 20307 USA. Walter Reed Army Med Ctr, Organ Transplant Serv, Washington, DC 20307 USA. Madigan Army Med Ctr, Serv Nephrol, Ft Lewis, WA USA. NIH, Minor Organ Tissue Transplant Educ Program, Bethesda, MD 20892 USA. Eisenhower Army Med Ctr, Serv Nephrol, Ft Gordon, GA USA. RP Abbott, KC (reprint author), Walter Reed Army Med Ctr, Dialysis Nephrol Serv, Washington, DC 20307 USA. OI Abbott, Kevin/0000-0003-2111-7112 NR 20 TC 20 Z9 20 U1 0 U2 1 PU WICHTIG EDITORE PI MILAN PA 72/74 VIA FRIULI, 20135 MILAN, ITALY SN 1121-8428 J9 J NEPHROL JI J. Nephrol. PD JUL-AUG PY 2001 VL 14 IS 4 BP 275 EP 279 PG 5 WC Urology & Nephrology SC Urology & Nephrology GA 458XD UT WOS:000170220000007 PM 11506250 ER PT J AU Lipsky, RH Xu, K Zhu, DM Kelly, C Terhakopian, A Novelli, A Marini, AM AF Lipsky, RH Xu, K Zhu, DM Kelly, C Terhakopian, A Novelli, A Marini, AM TI Nuclear factor kappa B is a critical determinant in N-methyl-D-aspartate receptor-mediated neuroprotection SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE gene expression; neuroprotection; NF-kappa B; brain-derived neurotrophic factor; NMDA receptors; oligonucleotide ID CEREBELLAR GRANULE NEURONS; INDUCED CELL-DEATH; NEUROTROPHIC FACTOR; HIPPOCAMPAL-NEURONS; INDUCED APOPTOSIS; OXIDATIVE STRESS; PRECURSOR P105; ACTIVATION; ALPHA; GLUTAMATE AB The role of a nuclear factor kappaB (NF-kappaB) in NMDA receptor-mediated neuroprotection is not known. A candidate sequence from the 5 ' flanking region of exon 3 of the rat brain-derived neurotrophic factor (BDNF) gene was used to show that exposure of rat cerebellar granule cells to 100 muM NMDA activated a specific DNA binding activity that was blocked by the NMDA receptor antagonist MK-801. Anti-p65 antibody or anti-p50 antibody 'supershifted' the DNA binding activity, suggesting that the DNA-protein complex was composed of p65 and p50 subunits. NMDA receptor-mediated neuroprotection was blocked when cerebellar neurons were transfected with a double-stranded oligonucleotide containing the BDNF gene NF-kappaB sequence. Furthermore, nuclear extracts prepared from neurons treated with NMDA and the double-stranded NF-kappaB oligonucleotide showed reduced DNA binding activity to the target sequence, supporting the idea that NF-kappaB may be involved in the transcriptional activation of the BDNF gene. To address this issue, we quantified the level of exon 3-specific BDNF mRNA. Relative to GAPDH mRNA levels and compared with untreated neurons, NMDA increased exon 3-specific BDNF mRNA twofold. In contrast, pretreatment of neurons with the NF-kappaB target DNA abolished the increase in BDNF mRNA following addition of NMDA, We also determined that BDNF itself induced an NF-kappaB DNA binding activity. Taken together, these data support a mechanism where NF-kappaB plays a critical role in NMDA-mediated neuroprotection. C1 NIAAA, Neurogenet Lab, NIH, Rockville, MD 20852 USA. Uniformed Serv Univ Hlth Sci, Dept Neurol, Bethesda, MD 20814 USA. Univ Oviedo, Fac Psychol, Dept Psychol, Oviedo, Spain. RP Lipsky, RH (reprint author), NIAAA, Neurogenet Lab, NIH, 12420 Parklawn Dr,Suite 451, Rockville, MD 20852 USA. RI Novelli, Antonello/P-7476-2015; OI Novelli, Antonello/0000-0002-0129-8350; Lipsky, Robert/0000-0001-7753-1473 NR 49 TC 116 Z9 127 U1 0 U2 4 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUL PY 2001 VL 78 IS 2 BP 254 EP 264 DI 10.1046/j.1471-4159.2001.00386.x PG 11 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 454NU UT WOS:000169979300006 PM 11461961 ER PT J AU Cardelli, P Fiori, A Corleto, VD Savi, MR Granata, F Ceci, F Ferraguti, G Potenza, RL Delle Fave, G Jensen, RT Strom, R AF Cardelli, P Fiori, A Corleto, VD Savi, MR Granata, F Ceci, F Ferraguti, G Potenza, RL Delle Fave, G Jensen, RT Strom, R TI Inhibitory effect of somatostatin on neutral amino acid transport in isolated brain microvessels SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE amino acid transport; blood-brain barrier; endothelium; somatostatin; somatostatin receptors ID RAT-BRAIN; BARRIER; RECEPTOR; PERMEABILITY; DEGRADATION; MEMBRANES; ANALOGS; PROTEIN; SYSTEMS; EFFLUX AB In the presence of somatostatin-14 or some of its receptorial agonists, the uptake of large neutral amino acids by isolated brain microvessels was found to be inhibited up to 50%, no other transport system being affected. Although the luminal and abluminal sides of brain endothelial cells are both capable of taking up large neutral amino acids, only uptake from the abluminal side appears to be inhibited by somatostatin. The involvement of a type-2 somatostatin receptor was suggested by assays with a series of receptor-specific somatostatin agonists, and was confirmed by the release of inhibition caused by a specific type-e receptor antagonist. A type-2-specific mRNA was indeed shown to be present in both bovine brain microvessels ex vivo and primary cultures of endothetial cells from rat brain microvessels. C1 Univ Rome La Sapienza, Dept Cellular Biotechnol & Haematol, I-00161 Rome, Italy. Univ Rome La Sapienza, Dept Biochem Sci A Rossi Fanelli, Rome, Italy. Univ Rome La Sapienza, Dept Cellular & Dev Biol, Rome, Italy. Univ Rome La Sapienza, Dept Clin Med, Gastroenterol Unit, Rome, Italy. NIDDK, Digest Dis Branch, NIH, Bethesda, MD USA. RP Strom, R (reprint author), Univ Rome La Sapienza, Dept Cellular Biotechnol & Haematol, Viale Regina Elena 324, I-00161 Rome, Italy. OI CECI, Fabrizio/0000-0003-4051-4922 NR 35 TC 2 Z9 2 U1 0 U2 2 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUL PY 2001 VL 78 IS 2 BP 349 EP 357 DI 10.1046/j.1471-4159.2001.00412.x PG 9 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 454NU UT WOS:000169979300015 PM 11461970 ER PT J AU Junn, E Mouradian, MM AF Junn, E Mouradian, MM TI Apoptotic signaling in dopamine-induced cell death: the role of oxidative stress, p38 mitogen-activated protein kinase, cytochrome c and caspases SO JOURNAL OF NEUROCHEMISTRY LA English DT Article DE apoptosis; caspase; cytochrome c; dopamine; p38 MAP kinase; Parkinson's disease ID CEREBELLAR GRANULE NEURONS; MITOCHONDRIAL COMPLEX-I; CENTRAL-NERVOUS-SYSTEM; PARKINSONS-DISEASE; PC12 CELLS; CASPASE-3-LIKE PROTEASES; MEDIATED APOPTOSIS; ALZHEIMERS-DISEASE; CYSTEINE PROTEASE; STRIATAL NEURONS AB Oxidative stress generated by dopamine (DA) oxidation could be one of the factors underlying the selective vulnerability of nigral dopaminergic neurons in Parkinson's diseases. Here we show that DA induces apoptosis in SH-SY5Y neuroblastoma cells demonstrated by activation of caspase-9 and caspase-3, cleavage of poly(ADP-ribose) polymerase as well as nuclear condensation. We also show that p38 mitogen-activated protein kinase is activated within 10 min of DA treatment, which precedes the onset of apoptosis because the potent p38 kinase inhibitor SB203580 protects against DA-induced cell death as well as against caspase-9 and caspase-3 activation. In addition, the antioxidant N-acetyl-L-cysteine (NAC) effectively blocks DA-induced p38 kinase activation, caspase-9 and caspase-3 cleavage and subsequent apoptosis, indicating that DA triggers apoptosis via a signaling pathway that is initiated by the generation of reactive oxygen species (ROS). Dopamine exerts its toxicity principally intracellularly as the DA uptake inhibitor, nomifensine significantly reduces DA-induced cell death as well as activation of p38 kinase and caspase-3. Furthermore, DA induces mitochondrial cytochrome c release, which is dependent on p38 kinase activation and precedes the cleavage of caspases. These observations indicate that DA induces apoptosis primarily by generating ROS, p38 kinase activation, cytochrome c release followed by caspase-9 and caspase-3 activation. C1 NINDS, Expt Therapeut Branch, Genet Pharmacol Unit, NIH, Bethesda, MD 20892 USA. RP Mouradian, MM (reprint author), NINDS, Expt Therapeut Branch, Genet Pharmacol Unit, NIH, 10 Ctr Dr,MSC 1406, Bethesda, MD 20892 USA. NR 74 TC 154 Z9 159 U1 0 U2 2 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0022-3042 J9 J NEUROCHEM JI J. Neurochem. PD JUL PY 2001 VL 78 IS 2 BP 374 EP 383 DI 10.1046/j.1471-4159.2001.00425.x PG 10 WC Biochemistry & Molecular Biology; Neurosciences SC Biochemistry & Molecular Biology; Neurosciences & Neurology GA 454NU UT WOS:000169979300018 PM 11461973 ER PT J AU Moore, DF Herscovitch, P Schiffmann, R AF Moore, DF Herscovitch, P Schiffmann, R TI Selective arterial distribution of cerebral hyperperfusion in Fabry disease SO JOURNAL OF NEUROIMAGING LA English DT Article DE Fabry disease; positron emission tomography; stroke; perfusion; cerebral blood flow ID HUMAN BRAIN; BLOOD-FLOW; TERRITORIES AB Fabry disease is an X-linked recessive deficiency of lysosomal alpha -galactosidase A associated with an increased risk of early onset cerebrovascular disease. The disorder is reported to affect the posterior circulation predominantly. This hypothesis was investigated directly by the measurement of regional cerebral blood flow with positron emission tomography (PET). Resting regional cerebral blood flow (rCBF) in 26 hemizygous patients with Fabry disease and 10 control participants was examined using (H2O)-O-15 and PET. Statistical parametric mapping (SPM{t}, SPM99) and PET images of patients and controls were produced. Significantly increased SPM{t} clusters were then color coded and blended with a coregistered T1 magnetic resonance imaging (MRI) template. Cerebral arterial territory maps were digitized and rescaled. Custom OpenGL and ImageVision Library C++ code was written to allow a first-order affine transformation of the blended SPM{t} and MRI template onto the arterial territory map. The affine transformation was constrained by choosing corresponding cerebral landmark "tie points" between the SPM{t} boolean OR MRI template images and the cerebral arterial territory maps. The data demonstrated that the posterior circulation is the predominant arterial territory with a significantly increased rCBF in Fabry disease. No arterial distribution had a decreased rCBF. C1 NINDS, Dev & Metab Neurol Branch, NIH, Bethesda, MD 20892 USA. NIH, Positron Emiss Tomog Dept, Bethesda, MD USA. RP Schiffmann, R (reprint author), NINDS, Dev & Metab Neurol Branch, NIH, Bldg 10,Room 3D03,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 13 TC 46 Z9 49 U1 1 U2 1 PU SAGE PUBLICATIONS INC PI THOUSAND OAKS PA 2455 TELLER RD, THOUSAND OAKS, CA 91320 USA SN 1051-2284 J9 J NEUROIMAGING JI J. Neuroimaging PD JUL PY 2001 VL 11 IS 3 BP 303 EP 307 PG 5 WC Clinical Neurology; Neuroimaging; Radiology, Nuclear Medicine & Medical Imaging SC Neurosciences & Neurology; Radiology, Nuclear Medicine & Medical Imaging GA 455FV UT WOS:000170017600013 PM 11462299 ER PT J AU Peng, YB Wu, J Willis, WD Kenshalo, DR AF Peng, YB Wu, J Willis, WD Kenshalo, DR TI GABA(A) and 5-HT3 receptors are involved in dorsal root reflexes: Possible role in periaqueductal gray descending inhibition SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID PRIMARY AFFERENT DEPOLARIZATION; RAT SPINAL-CORD; CENTRAL-NERVOUS-SYSTEM; BINDING-SITES; HORN NEURONS; AMINO-ACIDS; ELECTRICAL-STIMULATION; PRESYNAPTIC INHIBITION; INTRADERMAL INJECTION; JOINT INFLAMMATION AB The dorsal root reflex (DRR) is a measure of the central excitability of presynaptic inhibitory circuits in the spinal cord. Activation of the periaqueductal gray (PAG), a center for descending inhibition of spinal cord nociceptive transmission, induces release of variety of neurotransmitters in the spinal cord, including GABA and serotonin (5-HT). GABA has been shown to be involved in generation of DRRs. In this study, pharmacological agents that influence DRRs and their possible mechanisms were investigated. DRRs were recorded in anesthetized rats from filaments teased from the cut central stump of the left L-4 or L-5 dorsal root, using a monopolar recording electrode. Stimulating electrodes were placed either on the left sciatic nerve or transcutaneously in the left foot. Animals were paralyzed and maintained by artificial ventilation. Drugs were applied topically to the spinal cord. A total of 64 units were recorded in 34 Sprague-Dawley rats. Peripheral receptive fields were found for nine of these units. In these units, DRRs were evoked by brush, pressure, and pinch stimuli. Nine units were tested for an effect of electrical stimulation in the periaqueductal gray on the DRRs. In eight cases, DRR responses were enhanced following PAG stimulation. The background activity was 4.2 +/- 1.9 spikes/s (mean +/- SE; range: 0-97.7; n = 57). The responses to agents applied to the spinal cord were (in spikes/s): artificial cerebrospinal fluid, 7.1 +/- 3.6 (range: 0-86.9; n = 25); 0.1 mM GABA, 16.8 +/- 8.7 (range: 0-191.0; n = 22); 1.0 mM GABA, 116.0 +/- 26.5 (range: 0.05-1001.2; n = 50); and 1.0 mM phenylbiguanide (PBG), 68.1 +/- 25.3 (range: 0-1,073.0; n = 49). Bicuculline (0.5 mM, n = 27) and ondansetron (1.0 mM, n = 10) blocked the GABA and PBG effects, respectively (P< 0.05). Significant cross blockade was also observed. It is concluded that GABA(A) receptors are likely to play a key role in the generation of DRRs, but that 5-HT3 receptors may also contribute. DRRs can be modulated by supraspinal mechanisms through descending systems. C1 Univ Texas, Med Branch, Dept Anat & Neurosci, Galveston, TX 77555 USA. Natl Inst Dent & Cranofacial Res, Pain & Neurosensory Mechanisms Branch, NIH, Bethesda, MD 20892 USA. NIH, Ctr Sci Review, Bethesda, MD 20892 USA. RP Willis, WD (reprint author), Univ Texas, Med Branch, Dept Anat & Neurosci, Galveston, TX 77555 USA. FU NINDS NIH HHS [NS-09743] NR 85 TC 24 Z9 26 U1 0 U2 0 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3077 J9 J NEUROPHYSIOL JI J. Neurophysiol. PD JUL PY 2001 VL 86 IS 1 BP 49 EP 58 PG 10 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 454CP UT WOS:000169955100003 PM 11431487 ER PT J AU Del Negro, CA Johnson, SM Butera, RJ Smith, JC AF Del Negro, CA Johnson, SM Butera, RJ Smith, JC TI Models of respiratory rhythm generation in the pre-Botzinger complex. III. Experimental tests of model predictions SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID STEM IN-VITRO; PACEMAKER NEURONS; MEDULLARY SLICES; PREBOTZINGER COMPLEX; INSPIRATORY NEURONS; NEONATAL-RAT; SPINAL-CORD; NETWORK; MODULATION; MECHANISMS AB We used the testable predictions of mathematical models proposed by Butera et al. to evaluate cellular, synaptic, and population-level components of the hypothesis that respiratory rhythm in mammals is generated in vitro in the pre-Botzinger complex (pre-BotC) by a heterogeneous population of pacemaker neurons coupled by fast excitatory synapses. We prepared thin brain stem slices from neonatal rats that capture the pre-BotC and maintain inspiratory-related motor activity in vitro. We recorded pacemaker neurons extracellularly and found: intrinsic bursting behavior that did not depend on Ca(2+) currents and persisted after blocking synaptic transmission; multistate behavior with transitions from quiescence to bursting and tonic spiking states as cellular excitability was increased via extracellular K(+) concentration ([K(+)](o)); a monotonic increase in burst frequency and decrease in burst duration with increasing [K(+)](o); heterogeneity among different cells sampled; and an increase in inspiratory burst duration and decrease in burst frequency by excitatory synaptic coupling in the respiratory network. These data affirm the basis for the network model, which is composed of heterogeneous pacemaker cells having a voltage-dependent burst-generating mechanism dominated by persistent Na(+) current (I(NaP)) and excitatory synaptic coupling that synchronizes cell activity. We investigated population-level activity in the pre-BotC using local "macropatch" recordings and confirmed these model predictions: pre-BotC activity preceded respiratory-related motor output by 100-400 ms, consistent with a heterogeneous pacemaker-cell population generating inspiratory rhythm in the pre-BotC; pre-BotC population burst amplitude decreased monotonically with increasing [K(+)](o) (while frequency increased), which can be attributed to pacemaker cell properties; and burst amplitude fluctuated from cycle to cycle after decreasing bilateral synaptic coupling surgically as predicted from stability analyses of the model. We conclude that the pacemaker cell and network models explain features of inspiratory rhythm generation in vitro. C1 Natl Inst Neurol Disorders & Stroke, Cellular & Syst Neurobiol Sect, Neural Control Lab, Bethesda, MD 20892 USA. Georgia Inst Technol, Sch Elect & Comp Engn, Inst Bioengn & Biosci, Atlanta, GA 30332 USA. RP Smith, JC (reprint author), 49 Convent Dr,Rm 3A50, Bethesda, MD 20892 USA. EM jsmith@helix.nih.gov RI Del Negro, Ciro/K-3451-2013; OI Butera, Robert/0000-0002-1806-0621 NR 38 TC 96 Z9 98 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 JUL PY 2001 VL 86 IS 1 BP 59 EP 74 PG 16 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 454CP UT WOS:000169955100004 PM 11431488 ER PT J AU Lupica, CR Bell, JA Hoffman, AF Watson, PL AF Lupica, CR Bell, JA Hoffman, AF Watson, PL TI Contribution of the hyperpolarization-activated current (I-h)to membrane potential and GABA release in hippocampal interneurons SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID THALAMIC RELAY NEURONS; VOLTAGE-CLAMP ANALYSIS; CURRENT I-H; CATION CURRENT; INWARD CURRENT; SYNAPTIC TRANSMISSION; PRESYNAPTIC TERMINALS; PACEMAKER CHANNELS; OPIOID INHIBITION; CA1 NEURONS AB Intrinsic GABAergic interneurons provide inhibitory input to the principal neurons of the hippocampus. The majority of interneurons located in stratum oriens (s.o.) of the CA1 region express the hyperpolarization-activated cation current known as I-h. In an effort to elucidate the role of this current in regulating the baseline excitability of these neurons and its participation in the regulation of the release of GABA onto CA1 pyramidal neurons, we utilized whole cell electrophysiological recordings from both populations of cells. In voltage-clamp experiments, hyperpolarization of the interneuron membrane initiated a large inward current with an estimated activation threshold of 51.6 +/- 7.6 mV and a half-maximal voltage of -73.0 +/- 7.0 mV. This current was blocked by bath application of the I-h inhibitors ZD 7288 (50 muM) or cesium (2 mM). Current-clamp experiments at the interneuron resting membrane potential (-61.3 +/- 1.2 mV) revealed a significant hyperpolarization, a decrease in the rate of spontaneous action potential discharge, an increase in the cellular input resistance, and the elimination of rebound afterdepolarizations during blockade of I-h with ZD 7288 (50 muM). The hyperpolarizing effect of ZD 7288 was also substantially larger in interneurons clamped near -80 mV using current injection through the pipette. In addition to neurons exhibiting I-h, recordings were obtained from a small population of s.o. interneurons that did not exhibit this current. These cells demonstrated resting membrane potentials that were significantly more negative (-73.6 +/- 5.5 mV) than those observed in neurons expressing I-h, suggesting that this current contributes to more depolarized membrane potentials in these cells. Recordings from postsynaptic pyramidal neurons demonstrated that blockade of I-h with ZD 7288 caused a substantial reduction (similar to 43%) in the frequency of spontaneous action potential-dependent inhibitory postsynaptic currents (IPSCs), without altering their average amplitude. However, miniature action-potential-independent IPSC frequency, amplitude, and decay kinetics were unaltered by ZD 7288. These data suggest that I-h is active at the resting membrane potential in s.o. interneurons and as a result contributes to the spontaneous activity of these cells and to the tonic inhibition of CA1 pyramidal neurons in the hippocampus. C1 Univ Arizona, Hlth Sci Ctr, Dept Pharmacol, Tucson, AZ 85724 USA. Natl Inst Drug Abuse, Intramural Res Program, Baltimore, MD 21224 USA. RP Lupica, CR (reprint author), Univ Arizona, Hlth Sci Ctr, Dept Pharmacol, Rm 545,Life Sci N,1501 N Campbell Ave, Tucson, AZ 85724 USA. RI Hoffman, Alexander/H-3035-2012 OI Hoffman, Alexander/0000-0002-2676-0628 FU NIDA NIH HHS [DA-07725] NR 46 TC 100 Z9 104 U1 2 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 JUL PY 2001 VL 86 IS 1 BP 261 EP 268 PG 8 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 454CP UT WOS:000169955100023 PM 11431507 ER PT J AU Jagadeesh, B Chelazzi, L Mishkin, M Desimone, R AF Jagadeesh, B Chelazzi, L Mishkin, M Desimone, R TI Learning increases stimulus salience in anterior inferior temporal cortex of the macaque SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID SHORT-TERM-MEMORY; VISUAL-SEARCH; CORPUS-CALLOSUM; INFEROTEMPORAL CORTEX; HIPPOCAMPAL-FORMATION; SELECTIVE ATTENTION; OBJECT RECOGNITION; PERIRHINAL CORTEX; SPATIAL ATTENTION; RHESUS MONKEY AB With experience, an object can become behaviorally relevant and thereby quickly attract our interest when presented in a visual scene. A likely site of these learning effects is anterior inferior temporal (aIT) cortex, where neurons are thought to participate in the filtering of irrelevant information out of complex visual displays. We trained monkeys to saccade consistently to one of two pictures in an array, in return for a reward. The array was constructed by pairing two stimuli, one of which elicited a good response from the cell when presented alone ("good" stimulus) and the other of which elicited a poor response ("poor" stimulus). The activity of aIT cells was recorded while monkeys learned to saccade to either the good or poor stimulus in the array. We found that neuronal responses to the array were greater (before the saccade occurred) when training reinforced a saccade to the good stimulus than when training reinforced a saccade to the poor stimulus. This difference was not present on incorrect trials, i.e., when saccades to the incorrect stimulus were made. Thus the difference in activity was correlated with performance. The response difference grew over the course of the recording session, in parallel with the improvement in performance. The response difference was not preceded by a difference in the baseline activity of the cells, unlike what was found in studies of cued visual search and working memory in aIT cortex. Furthermore, we found similar effects in a version of the task in which any of 10 possible pairs of stimuli, prelearned before the recording session, could appear on a given trial, thereby precluding a working memory strategy. The results suggest that increasing the behavioral significance of a stimulus through training alters the neural representation of that stimulus in aIT cortex. As a result, neurons responding to features of the relevant stimulus may suppress neurons responding to features of irrelevant stimuli. C1 Univ Washington, Dept Physiol & Biophys, Seattle, WA 98195 USA. NIMH, Neuropsychol Lab, Bethesda, MD 20892 USA. Univ Verona, Dipartimento Sci Neurol & Vis, Sez Fisiol, I-37134 Verona, Italy. RP Jagadeesh, B (reprint author), Univ Washington, Dept Physiol & Biophys, Box 357330, Seattle, WA 98195 USA. RI Chelazzi, Leonardo/B-6084-2011 OI Chelazzi, Leonardo/0000-0001-8566-0611 NR 46 TC 58 Z9 58 U1 1 U2 5 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3077 J9 J NEUROPHYSIOL JI J. Neurophysiol. PD JUL PY 2001 VL 86 IS 1 BP 290 EP 303 PG 14 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 454CP UT WOS:000169955100026 PM 11431510 ER PT J AU Burke, RE Degtyarenko, AM Simon, ES AF Burke, RE Degtyarenko, AM Simon, ES TI Patterns of locomotor drive to motoneurons and last-order interneurons: Clues to the structure of the CPG SO JOURNAL OF NEUROPHYSIOLOGY LA English DT Article ID DISTAL HINDLIMB MUSCULATURE; BACKWARD QUADRUPEDAL WALKING; LATENCY CUTANEOUS EXCITATION; FLEXOR DIGITORUM LONGUS; PHASIC GAIN-CONTROL; FICTIVE LOCOMOTION; ALPHA-MOTONEURONES; REFLEX PATHWAYS; MOTOR PATTERNS; LUMBOSACRAL MOTONEURONS AB We have examined the linkage between patterns of activity in several hindlimb motor pools and the modulation of oligosynaptic cutaneous reflex pathways during fictive locomotion in decerebrate unanesthetized cats to assess the notion that such linkages can shed light on the structure of the central pattern generator (CPG) for locomotion. We have concentrated attention on the cutaneous reflex pathways that project to the flexor digitorum longus (FDL) motor pool because of that muscle's unique variable behavior during normal and fictive locomotion in the cat. Differential locomotor control of last-order excitatory interneurons in pathways from low-threshold cutaneous afferents in the superficial peroneal and medial plantar afferents to FDL motoneurons is fully documented for the first time. The qualitative patterns of differential control are shown to remain the same whether the FDL muscle is active in early flexion, as usually found, or during the extension phase of fictive locomotion, which is less common during fictive stepping. The patterns of motor pool activity and of reflex pathway modulation indicate that the flexion phase of fictive locomotion has distinct early versus late components. Observations during "normal" and unusual patterns of fictive stepping suggest that some aspects of locomotor pattern formation can be separated from rhythm generation, implying that these two CPG functions may be embodied, at least in part, in distinct neural organizations. The results are discussed in relation to a provisional circuit diagram that could explain the experimental findings. C1 NINDS, Neural Control Lab, NIH, Bethesda, MD 20892 USA. RP Burke, RE (reprint author), NINDS, Neural Control Lab, NIH, Bldg 49,Rm 3A50, Bethesda, MD 20892 USA. EM reburke@helix.nih.gov RI X, Simon/F-4678-2011 NR 64 TC 86 Z9 87 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 JUL PY 2001 VL 86 IS 1 BP 447 EP 462 PG 16 WC Neurosciences; Physiology SC Neurosciences & Neurology; Physiology GA 454CP UT WOS:000169955100040 PM 11431524 ER PT J AU Pyrski, M Xu, Z Walters, E Gilbert, DJ Jenkins, NA Copeland, NG Margolis, FL AF Pyrski, M Xu, Z Walters, E Gilbert, DJ Jenkins, NA Copeland, NG Margolis, FL TI The OMP-lacZ transgene mimics the unusual expression pattern of OR-Z6, a new odorant receptor gene on mouse chromosome 6: Implication for locus-dependent gene expression SO JOURNAL OF NEUROSCIENCE LA English DT Article DE transgene; receptors; OR-Z6; olfactory neurons; olfactory epithelium; olfactory bulb; expression patterns; in situ hybridization; locus dependence; OMP promoter ID PUTATIVE PHEROMONE RECEPTORS; OLFACTORY-BULB; MULTIGENE FAMILY; TOPOGRAPHIC MAP; NEURONS; ORGANIZATION; TRANSCRIPTION; MAMMALS; MICE; GLOMERULI AB Reporter gene expression in the olfactory epithelium of H-lacZ6 transgenic mice mimics the cell-selective expression pattern known for some odorant receptor genes. The transgene construct in these mice consists of the lacZ coding region, driven by the proximal olfactory marker protein (OMP) gene promoter, and shows expression in a zonally confined subpopulation of olfactory neurons. To address mechanisms underlying the odorant receptor-like expression pattern of the lacZ construct, we analyzed the transgene-flanking region and identified OR-Z6, the first cloned odorant receptor gene that maps to mouse chromosome 6. OR-Z6 bears the highest sequence similarity (85%) to a human odorant receptor gene at the syntenic location on human chromosome 7. We analyzed the expression pattern of OR-Z6 in olfactory tissues of H-lacZ6 mice and show that it bears strong similarities to that mapped for beta -galactosidase. Expression of both genes in olfactory neurons is primarily restricted to the same medial subregion of the olfactory epithelium. Axons from both neuronal subpopulations project to the same ventromedial aspect of the anterior olfactory bulbs. Furthermore, colocalization analyses in H-lacZ6 mice demonstrate that OR-Z6-reactive glomeruli receive axonal input from lacZ-positive neurons as well. These results suggest that the expression of both genes is coordinated and that transgene expression in H-lacZ6 mice is regulated by locus-dependent mechanisms. C1 Univ Maryland, Sch Med, Dept Anat & Neurobiol, Hlth Sci Facil 273, Baltimore, MD 21201 USA. Howard Univ, Coll Med, Dept Biochem & Mol Biol, Washington, DC 20059 USA. NCI, Mouse Canc Genet Program, Frederick, MD 21702 USA. RP Margolis, FL (reprint author), Univ Maryland, Sch Med, Dept Anat & Neurobiol, Hlth Sci Facil 273, 685 W Baltimore St, Baltimore, MD 21201 USA. FU NIDCD NIH HHS [DCD03112]; NINDS NIH HHS [U54NS39407] NR 52 TC 36 Z9 36 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 JUL 1 PY 2001 VL 21 IS 13 BP 4637 EP 4648 PG 12 WC Neurosciences SC Neurosciences & Neurology GA 445JP UT WOS:000169454600012 PM 11425891 ER PT J AU Iuliano, BA Nguyen, TT Fuller, BG Schrager, JA Oldfield, EH AF Iuliano, BA Nguyen, TT Fuller, BG Schrager, JA Oldfield, EH TI Cerebrospinal fluid rhinorrhea caused by metastatic angiosarcoma - Case illustration SO JOURNAL OF NEUROSURGERY LA English DT Editorial Material DE cerebrospinal fluid rhinorrhea; angiosarcoma; cisternography; transsphenoidal approach C1 NINCDS, Surg Neurol Branch, NIH, Bethesda, MD 20892 USA. Mayo Clin & Mayo Fdn, Dept Neurol Surg, Rochester, MN 55905 USA. NCI, Radiat Oncol Branch, NIH, Bethesda, MD 20892 USA. NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. RP Oldfield, EH (reprint author), NINCDS, Surg Neurol Branch, NIH, Bldg 10,Room 5D37, Bethesda, MD 20892 USA. NR 5 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC NEUROLOGICAL SURGEONS PI CHARLOTTESVILLE PA UNIV VIRGINIA, 1224 WEST MAIN ST, STE 450, CHARLOTTESVILLE, VA 22903 USA SN 0022-3085 J9 J NEUROSURG JI J. Neurosurg. PD JUL PY 2001 VL 95 IS 1 BP 143 EP 143 DI 10.3171/jns.2001.95.1.0143 PG 1 WC Clinical Neurology; Surgery SC Neurosciences & Neurology; Surgery GA 448JC UT WOS:000169622900023 PM 11453386 ER PT J AU Oldfield, EH AF Oldfield, EH TI Syringomyelia SO JOURNAL OF NEUROSURGERY LA English DT Editorial Material ID SPINAL-CORD; FLUID-FLOW; PATHOPHYSIOLOGY C1 NIH, Dept Hlth & Human Serv, Bethesda, MD 20892 USA. RP Oldfield, EH (reprint author), NIH, Dept Hlth & Human Serv, Bldg 10, Bethesda, MD 20892 USA. NR 22 TC 10 Z9 10 U1 0 U2 0 PU AMER ASSOC NEUROLOGICAL SURGEONS PI CHARLOTTESVILLE PA UNIV VIRGINIA, 1224 WEST MAIN ST, STE 450, CHARLOTTESVILLE, VA 22903 USA SN 0022-3085 J9 J NEUROSURG JI J. Neurosurg. PD JUL PY 2001 VL 95 IS 1 SU S BP 153 EP 155 DI 10.3171/spi.2001.95.1.0153 PG 3 WC Clinical Neurology; Surgery SC Neurosciences & Neurology; Surgery GA 449ME UT WOS:000169688300032 PM 11453422 ER PT J AU Hoffman, JM Coleman, RE AF Hoffman, JM Coleman, RE TI SPECT imaging in dementias - Reply SO JOURNAL OF NUCLEAR MEDICINE LA English DT Letter ID BRAIN BLOOD-FLOW; HISTOPATHOLOGIC CORRELATION C1 NCI, Bethesda, MD 20892 USA. Duke Univ, Durham, NC 27706 USA. RP Hoffman, JM (reprint author), NCI, Bethesda, MD 20892 USA. NR 3 TC 0 Z9 0 U1 0 U2 0 PU SOC NUCLEAR MEDICINE INC PI RESTON PA 1850 SAMUEL MORSE DR, RESTON, VA 20190-5316 USA SN 0161-5505 J9 J NUCL MED JI J. Nucl. Med. PD JUL PY 2001 VL 42 IS 7 BP 1132 EP 1133 PG 2 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 447QB UT WOS:000169583000039 ER PT J AU Weinstein, SJ Ziegler, RG Frongillo, EA Colman, N Sauberlich, HE Brinton, LA Hamman, RF Levine, RS Mallin, K Stolley, PD Bisogni, CA AF Weinstein, SJ Ziegler, RG Frongillo, EA Colman, N Sauberlich, HE Brinton, LA Hamman, RF Levine, RS Mallin, K Stolley, PD Bisogni, CA TI Low serum and red blood cell folate are moderately, but nonsignificantly associated with increased risk of invasive cervical cancer in US women SO JOURNAL OF NUTRITION LA English DT Article DE cervix neoplasms; serum folate; red blood cell folate; microbiologic folate assay; radiobinding folate assay; humans ID HUMAN PAPILLOMAVIRUS; UNITED-STATES; FOLIC-ACID; METHYLENETETRAHYDROFOLATE REDUCTASE; INTRAEPITHELIAL NEOPLASIA; ORAL-CONTRACEPTIVES; WHITE WOMEN; DYSPLASIA; HOMOCYSTEINE; DIET AB Previous observational epidemiologic studies of folate and cervical cancer, as well as folate supplementation trials for cervical dysplasia, have produced mixed results. We examined the relationship between serum and RBC folate and incident invasive cervical cancer in a large, multicenter, community-based case-control study. Detailed in-person interviews were conducted, and blood was drawn at least 6 mo after completion of cancer treatment from 51% of cases and 68% of controls who were interviewed. Blood folate was measured with both microbiologic and radiobinding assays. Included in the final analyses were 183 cases and 540 controls. Logistic regression was used to control for all accepted risk factors, including age, sexual behavior, smoking, oral contraceptive use, Papanicolaou smear history and human papillomavirus (HPV)-16 serology. For all four folate measures, the geometric mean in cases was lower than in controls (e.g., 11.6 vs. 13.0 nmol/L, P < 0.01 for the serum radiobinding assay). Folate measures using microbiologic and radiobinding assays were correlated (serum: r = 0.90; RBC: r = 0.72). For serum folate, multivariate-adjusted odds ratios (OR) in the lowest vs. highest quartile were 1.3 [95% confidence interval (CI) = 0.8-2.9] and 1.6 (0.9-2.9), using the microbiologic and radiobinding assays, respectively. For RBC folate, comparable OR were 1.2 (0.6-2.2) and 1.5 (0.8-2.7). Similar risks were obtained when restricting analyses to subjects with a history of HPV infection. Thus, low serum and RBC folate were each moderately, but nonsignificantly, associated with increased invasive cervical cancer risk. These findings support a role for one-carbon metabolism in the etiology of cervical cancer. C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Cornell Univ, Div Nutr Sci, Ithaca, NY 14853 USA. Columbia Univ, St Lukes Roosevelt Hosp Ctr, Dept Pathol & Lab Med, New York, NY 10025 USA. Columbia Univ, New York, NY 10025 USA. Univ Alabama, Dept Nutr Sci, Birmingham, AL 35294 USA. Univ Colorado, Hlth Sci Ctr, Dept Prevent Med & Biometr, Denver, CO 80262 USA. Meharry Med Coll, Sch Med Occupat & Prevent Med, Nashville, TN 37208 USA. Univ Illinois, Sch Publ Hlth, Div Epidemiol & Biostat, Chicago, IL 60612 USA. Univ Maryland, Sch Med, Dept Epidemiol & Prevent Med, Baltimore, MD 21201 USA. RP Ziegler, RG (reprint author), NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. RI Brinton, Louise/G-7486-2015 OI Brinton, Louise/0000-0003-3853-8562 NR 65 TC 17 Z9 21 U1 0 U2 10 PU AMER INST NUTRITION PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-3166 J9 J NUTR JI J. Nutr. PD JUL PY 2001 VL 131 IS 7 BP 2040 EP 2048 PG 9 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 452BH UT WOS:000169836900028 PM 11435527 ER PT J AU Zheng, TZ Zahm, SH Cantor, KP Weisenburger, DD Zhang, YW Blair, A AF Zheng, TZ Zahm, SH Cantor, KP Weisenburger, DD Zhang, YW Blair, A TI Agricultural exposure to carbamate pesticides and risk of non-Hodgkin lymphoma SO JOURNAL OF OCCUPATIONAL AND ENVIRONMENTAL MEDICINE LA English DT Article ID MICE; GROUNDWATER; CARBOFURAN; MINNESOTA; IOWA; DDT; MEN AB Recent epidemiological studies have suggested an increased risk of non-Hodgkin lymphoma (NHL)Srs m carbamate insecticide use among farmers. To further explore the possible relationships, we conducted a pooled analysis of three population-based case-control studies conducted in four midwestern states in the United States. A total of 985 white male subjects and 2895 control subjects were included in this analysis. Unconditional logistic regression was used to estimate the association and control for confounding. Compared with nonfarmers, farmers who had ever used carbamate pesticides had a 30 % to 50% increased risk of NHL, whereas farmers without carbamate pesticide use showed no increased risk. Analyses for individual carbamate pesticides found a more consistent association with Sevin but not carbofuran, butylate, or S-ethyl dipropylthiocarbamate plus protectant, Among farmers using Satin, the risk of NHL was limited to those who personally handled the product, those who first used the product for greater than or equal to 20 years before their disease diagnosis, and those who used the product for a longer period. These associations persisted after adjusting for other major classes of pesticides. These results suggest an increased risk of NHL associated with carbamate pesticide use, particularly Sevin. Further investigation of the association is warranted. C1 Yale Univ, Sch Publ Hlth, New Haven, CT 06520 USA. NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Univ Nebraska, Med Ctr, Lincoln, NE 68583 USA. RP Zheng, TZ (reprint author), Suite 700-703,129 Church St, New Haven, CT 06510 USA. NR 33 TC 73 Z9 77 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 1076-2752 J9 J OCCUP ENVIRON MED JI J. Occup. Environ. Med. PD JUL PY 2001 VL 43 IS 7 BP 641 EP 649 DI 10.1097/00043764-200107000-00012 PG 9 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 451VQ UT WOS:000169823400012 PM 11464396 ER PT J AU Nocito, A Bubendorf, L Tinner, EM Suess, K Wagner, U Forster, T Kononen, J Fijan, A Bruderer, J Schmid, U Ackermann, D Maurer, R Alund, G Knonagel, H Rist, M Anabitarte, M Hering, F Hardmeier, T Schoenenberger, AJ Flury, R Jager, P Fehr, JL Schraml, P Moch, H Mihatsch, MJ Gasser, T Sauter, G AF Nocito, A Bubendorf, L Tinner, EM Suess, K Wagner, U Forster, T Kononen, J Fijan, A Bruderer, J Schmid, U Ackermann, D Maurer, R Alund, G Knonagel, H Rist, M Anabitarte, M Hering, F Hardmeier, T Schoenenberger, AJ Flury, R Jager, P Fehr, JL Schraml, P Moch, H Mihatsch, MJ Gasser, T Sauter, G TI Microarrays of bladder cancer tissue are highly representative of proliferation index and histological grade SO JOURNAL OF PATHOLOGY LA English DT Article DE bladder cancer; tissue microarrays; Ki67 LI; grade; prognosis; TMA ID S-PHASE FRACTION; URINARY-BLADDER; CLINICAL-SIGNIFICANCE; PROGNOSTIC FACTORS; PROSTATE-CANCER; CDNA MICROARRAY; CELL CARCINOMA; DNA-PLOIDY; EXPRESSION; TUMORS AB The number of genes suggested to play a role in cancer biology is rapidly increasing, To be able to test a large number of molecular parameters in sufficiently large series of primary tumours, a tissue microarray (TMA) approach has been developed where samples from up to 1000 tumours can be simultaneously analysed on one glass slide. Because of the small size of the individual arrayed tissue samples (diameter 0.6 mm), the question arises of whether these specimens are representative of their donor tumours. To investigate how representative are the results obtained on TMAs, a set of 2317 bladder tumours that had been previously analysed for histological grade and Ki67 labelling index (LI) was used to construct four replica TMAs from different areas of each tumour. Clinical follow-up information was available from 1092 patients. The histological grade and the Ki67 LI were determined fbr every arrayed tumour sample (4 x 2317 analyses each). Despite discrepancies in individual cases, the grade and Ki67 information obtained on minute arrayed samples were highly similar to the data obtained on large sections (p <0.0001). Most importantly, every individual association between grade or Ki67 LI and tumour stage or prognosis (recurrence, progression, tumour-specific survival) that was observed in large section analysis could be fully reproduced on all four replica TMAs, These results show that intra-tumour heterogeneity does not significantly affect the ability to detect clinico-pathological correlations on TMAs, probably because of the large number of tumours that can be included in TMA studies. TMAs are a powerful tool for rapid identification of the biological or clinical significance of molecular alterations in bladder cancer and other tumour types. Copyright (C) 2001 John Wiley & Sons, Ltd. C1 Univ Basel, Inst Pathol, CH-4003 Basel, Switzerland. Cantonal Hosp Schaffhausen, Urol Clin, Schaffhausen, Switzerland. Cantonal Hosp Winterthur, Urol Clin, Winterthur, Switzerland. Cantonal Hosp Winterthur, Inst Pathol, Winterthur, Switzerland. Cantonal Hosp Munsterlingem, Urol Clin, Munsterlingem, Switzerland. Cantonal Hosp Munsterlingem, Inst Pathol, Munsterlingem, Switzerland. Cantonal Hosp Baden, Urol Clin, Baden, Switzerland. Clara Hosp, Basel, Switzerland. Limmattal Hosp, Urol Clin, Schlieren, Switzerland. City Hosp Triemli, Inst Pathol, Zurich, Switzerland. Cantonal Hosp St Gallen, Urol Clin, St Gallen, Switzerland. Cantonal Hosp St Gallen, Inst Pathol, St Gallen, Switzerland. NHGRI, Canc Genet Lab, NIH, Bethesda, MD 20892 USA. Univ Basel, Urol Clin, CH-4003 Basel, Switzerland. RP Sauter, G (reprint author), Univ Basel, Inst Pathol, Schoenbeinstr 40, CH-4003 Basel, Switzerland. RI Bubendorfl, Lukas/H-5880-2011 NR 26 TC 234 Z9 241 U1 2 U2 8 PU JOHN WILEY & SONS LTD PI W SUSSEX PA BAFFINS LANE CHICHESTER, W SUSSEX PO19 1UD, ENGLAND SN 0022-3417 J9 J PATHOL JI J. Pathol. PD JUL PY 2001 VL 194 IS 3 BP 349 EP 357 DI 10.1002/1096-9896(200107)194:3<349::AID-PATH887>3.0.CO;2-D PG 9 WC Oncology; Pathology SC Oncology; Pathology GA 448PJ UT WOS:000169635600011 PM 11439368 ER PT J AU Van Dyke, T Ryder, M Cordero, R Gallagher, A Griffiths, G Offenbacher, S Cizza, G Kimberlin, M Lombardi, A Meng, L Shih, W AF Van Dyke, T Ryder, M Cordero, R Gallagher, A Griffiths, G Offenbacher, S Cizza, G Kimberlin, M Lombardi, A Meng, L Shih, W CA Alendronate Periodontal Study Grp TI Safety of once-weekly alendronate 70 mg in periodontal disease SO JOURNAL OF RHEUMATOLOGY LA English DT Meeting Abstract C1 Boston Univ, Boston, MA 02215 USA. Univ Calif San Francisco, San Francisco, CA 94143 USA. Hilltop Res, W Palm Beach, FL USA. Hilltop Res, Miamiville, OH USA. Scott & White Mem Hosp & Clin, Temple, TX 76508 USA. Univ N Carolina, Chapel Hill, NC USA. NIH, Bethesda, MD 20892 USA. Merck Res Labs, Rahway, NJ USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU J RHEUMATOL PUBL CO PI TORONTO PA 920 YONGE ST, SUITE 115, TORONTO, ONTARIO M4W 3C7, CANADA SN 0315-162X J9 J RHEUMATOL JI J. Rheumatol. PD JUL PY 2001 VL 28 SU 63 MA M93 BP 26 EP 26 PG 1 WC Rheumatology SC Rheumatology GA 452ZH UT WOS:000169889200095 ER PT J AU Naitow, H Canady, MA Lin, TW Wickner, RB Johnson, JE AF Naitow, H Canady, MA Lin, TW Wickner, RB Johnson, JE TI Purification, crystallization, and preliminary X-ray analysis of L-A: A dsRNA yeast virus SO JOURNAL OF STRUCTURAL BIOLOGY LA English DT Article DE X-ray crystallography; crystallization; diffraction; L-A virus; Totivirus; Totiviridae ID DOUBLE-STRANDED-RNA; MESSENGER-RNA; DEGRADATION; REPLICATION; CORE AB The L-A virus (LAV) particle is a specialized compartment for the transcription and replication of double-stranded RNA. It is 390 Angstrom in diameter and infects yeast. The particle is formed by a capsid containing 120 copies of a 680-residue gene product arranged with T = I icosahedral symmetry, approximately two copies of an RNA-directed RNA polymerase, and a 4.6-kb linear, duplex RNA. LAV crystals diffracting to at least 4.5-Angstrom resolution were grown in a combination of polyethylene glycol 8000, ethylene glycol, and lithium chloride. Following crystallization the reservoir solution was replaced by a 2 X concentrated reservoir solution in order for ethylene glycol to function as a cryoprotectant even though initial crystals would not grow at sufficiently high concentrations of ethylene glycol for cryoprotection. A complete data set was collected to 6-Angstrom resolution from a frozen crystal obtained with this procedure. The crystals belong to space group P2(1). The unit cell dimensions are a = 406.7 Angstrom, b = 403.3 Angstrom, c = 572.5 Angstrom, beta = 90.3 degrees with two virus particles in the unit cell. The particle orientation was determined with the rotation function and the particle center was estimated on the basis of packing considerations. (C) 2001 Academic Press. C1 Scripps Res Inst, Dept Mol Biol, La Jolla, CA 92037 USA. NIDDK, Lab Biochem & Genet, NIH, Bethesda, MD 20892 USA. RP Johnson, JE (reprint author), Scripps Res Inst, Dept Mol Biol, 10550 N Torrey Pines Rd,MB-31, La Jolla, CA 92037 USA. FU NCRR NIH HHS [RR-07707]; NIGMS NIH HHS [GM54076] NR 11 TC 8 Z9 8 U1 1 U2 1 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1047-8477 J9 J STRUCT BIOL JI J. Struct. Biol. PD JUL PY 2001 VL 135 IS 1 BP 1 EP 7 DI 10.1006/jsbi.2001.4371 PG 7 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 480DU UT WOS:000171447400001 PM 11562160 ER PT J AU Kim, HC Nemes, Z Idler, WW Hyde, CC Steinert, PM Ahvazi, B AF Kim, HC Nemes, Z Idler, WW Hyde, CC Steinert, PM Ahvazi, B TI Crystallization and preliminary X-ray analysis of human transglutaminase 3 from zymogen to active form SO JOURNAL OF STRUCTURAL BIOLOGY LA English DT Article DE transglutaminase 3; crystallization; twinning; X-ray crystallography ID CONGENITAL ICHTHYOSIFORM ERYTHRODERMA; CORNIFIED CELL-ENVELOPE; LAMELLAR ICHTHYOSIS; CROSS-LINKING; KERATINOCYTE TRANSGLUTAMINASE; PROTRANSGLUTAMINASE-E; INCREASED EXPRESSION; PROTEINS; MUTATIONS; DEATH AB Transglutaminases (TGases; protein-glutamine-glutamyl-transferases) are a large family of calcium-dependent acyl-transfer enzymes that catalyze the formation of covalent cross links in proteins. Of these, the "epidermal" or "hair follicle" TGase 3 isoform is critically involved in barrier formation in epithelia. It is a zymogen, requiring proteolytic activation to achieve maximal specific activity. In order to understand its structure and function, we have devised methods for the rapid large-scale expression of the TGase 3 zymogen in the baculovirus system, and here we describe the purification of the zymogen and activated forms. We describe methods for the formation of high-quality, well-diffracting crystals within 3-5 days, using both dioxane and beta -octylglucoside to overcome severe twinning problems. The crystal of the zymogen belongs to the triclinic space group P1 and diffracts to 2.2-Angstrom resolution, and the crystal of the active form belongs to the P2(1) space group at 2.7-Angstrom resolution. C1 NIAMSD, Skin Biol Lab, NIH, Bethesda, MD 20892 USA. NIAMSD, Struct Biol Res Lab, NIH, Bethesda, MD 20892 USA. RP Ahvazi, B (reprint author), NIAMSD, Skin Biol Lab, NIH, Bldg 6,Room B2-16,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 30 TC 13 Z9 13 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1047-8477 J9 J STRUCT BIOL JI J. Struct. Biol. PD JUL PY 2001 VL 135 IS 1 BP 73 EP 77 DI 10.1006/jsbi.2001.4384 PG 5 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 480DU UT WOS:000171447400009 PM 11562168 ER PT J AU Smothers, B Bertolucci, D AF Smothers, B Bertolucci, D TI Alcohol consumption and health-promoting behavior in a US household sample: Leisure-time physical activity SO JOURNAL OF STUDIES ON ALCOHOL LA English DT Article ID CORONARY HEART-DISEASE; CARDIOVASCULAR-DISEASE; UNITED-STATES; RISK-FACTORS; MORTALITY; SMOKING; MEN; LIPOPROTEINS; POPULATION; HABITS AB Objective: Hypothesizing that people who moderate their alcohol intake engage in other health-promoting cardioprotective behavior, this research explored associations between alcohol consumption and leisure-time physical activity. Method: Data representative of the U.S. population (N = 41,104; 52.5% women) were derived from the 1990 National Health Interview Survey, which employed a multistage probability design. Multivariate logistic regression analyses were performed, using lifetime abstainers as the reference group. Results: An inverted J-shaped curve characterized the association between estimates of alcohol consumption and leisure-time physical activity, in which likelihood of displaying a physically active lifestyle (odds ratios) increased from abstinence (1.00) to moderate drinking (1.84), then declined at heavier consumption (1.61). Stratified analyses yielded similar results with peaks at light or moderate consumption for current smokers, persons aged 55 and over, and those with cardiovascular risk conditions. As activity intensity increased, the inverted J-shaped curve persisted with more pronounced peaks at moderate drinking. Conclusions: Robust findings of an association between cardioprotective leisure-time physical activity and moderate alcohol consumption provide evidence for a health-promoting lifestyle that may play an explanatory role in the alcohol-coronary heart disease relationship. C1 NIAAA, Epidemiol Branch, Div Biometry & Epidemiol, NIH, Bethesda, MD 20892 USA. RP Smothers, B (reprint author), NIAAA, Epidemiol Branch, Div Biometry & Epidemiol, NIH, Willco Bldg,Suite 514,6000 Execut Blvd, Bethesda, MD 20892 USA. NR 36 TC 28 Z9 28 U1 2 U2 4 PU ALCOHOL RES DOCUMENTATION INC CENT ALCOHOL STUD RUTGERS UNIV PI PISCATAWAY PA C/O DEIRDRE ENGLISH, 607 ALLISON RD, PISCATAWAY, NJ 08854-8001 USA SN 0096-882X J9 J STUD ALCOHOL JI J. Stud. Alcohol PD JUL PY 2001 VL 62 IS 4 BP 467 EP 476 PG 10 WC Substance Abuse; Psychology SC Substance Abuse; Psychology GA 461DD UT WOS:000170348900007 PM 11513224 ER PT J AU Kissin, WB Svikis, DS Morgan, GD Haug, NA AF Kissin, WB Svikis, DS Morgan, GD Haug, NA TI Characterizing pregnant drug-dependent women in treatment and their children SO JOURNAL OF SUBSTANCE ABUSE TREATMENT LA English DT Article DE pregnant women; drug treatment; treatment needs; substance abuse; child custody ID ADDICTION SEVERITY INDEX; SUBSTANCE-ABUSERS; ALCOHOL AB Alcohol and other drug use during pregnancy represents a major public health concern. This article characterizes a sample of 240 pregnant opioid- or cocaine-dependent women enrolled in the initial residential component of a comprehensive substance abuse treatment program for pregnant women. Data were collected using the Addiction Severity Index, Psychosocial History Form, and Structured Clinical Interview for DSM-III-R. Patients' substance use history, psychiatric comorbidity, social support network, employability, current and previous pregnancies, child custody arrangements, and the father of the current pregnancy, are described to assist providers in tailoring treatment to the specific needs of this population. (C) 2001 Elsevier Science Inc. All rights reserved. C1 WESTAT Corp, Rockville, MD 20850 USA. Virginia Commonwealth Univ, Dept Psychol, Richmond, VA 23284 USA. NCI, Bethesda, MD 20892 USA. Johns Hopkins Univ, Sch Med, Dept Psychiat, Baltimore, MD 21205 USA. RP Kissin, WB (reprint author), WESTAT Corp, 1650 Res Blvd, Rockville, MD 20850 USA. FU NIDA NIH HHS [P50 5T32DA 07209, P50 DA 09258] NR 22 TC 36 Z9 36 U1 1 U2 3 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0740-5472 J9 J SUBST ABUSE TREAT JI J. Subst. Abus. Treat. PD JUL PY 2001 VL 21 IS 1 BP 27 EP 34 DI 10.1016/S0740-5472(01)00176-3 PG 8 WC Psychology, Clinical; Substance Abuse SC Psychology; Substance Abuse GA 467TR UT WOS:000170720300004 PM 11516924 ER PT J AU Liu, XC Sun, ZX Neiderhiser, JM Uchiyama, M Okawa, M Rogan, W AF Liu, XC Sun, ZX Neiderhiser, JM Uchiyama, M Okawa, M Rogan, W TI Behavioral and emotional problems in Chinese adolescents: Parent and teacher reports SO JOURNAL OF THE AMERICAN ACADEMY OF CHILD AND ADOLESCENT PSYCHIATRY LA English DT Article DE behavioral problems; child behavior checklist; emotional disorder ID CHILD PSYCHIATRIC EPIDEMIOLOGY; AGED 6; INFORMANTS; AGREEMENT; DEPRESSION; PREVALENCE; COMORBIDITY; INTERVIEW; SYMPTOMS; MOTHERS AB Objective: This study examined behavioral and emotional problems in Chinese adolescents. Method: A sample of 1,694 adolescents aged 12 to 16 years participated in this study in 1997. Parents completed the Child Behavior Checklist (CBCL), and teachers completed the Teacher's Report Form (TRF). Results: For both parent and teacher reports. internalizing syndromes were scored higher in girls and externalizing syndromes were scored higher in boys. Scores on most of the CBCL and TRF subscales were higher for the older adolescents. The overall prevalence rates of parent- and teacher-reported behavioral problems were 23.1% and 19.2%, respectively. The eight cross-informant syndromes were highly comorbid, with a significant association across syndromes (mean odds ratio = 16.1 for CBCL and 22.5 for TRF). Correlations between parent and teacher reports were 0.51 for eight subscales and 0.68 for Total Problems. Conclusions: These findings demonstrate that behavioral and emotional problems tend to increase with age and cluster in the same individuals. The prevalence rates of behavioral problems in Chinese adolescents are comparable to those reported in Western countries. In contrast to findings for Western samples, parent reports of behavioral problems were highly correlated with teacher reports. C1 NIEHS, Epidemiol Branch, Res Triangle Pk, NC 27709 USA. Shandong Univ, Dept Psychiat, Shandong, Peoples R China. George Washington Univ, Washington, DC USA. RP Liu, XC (reprint author), NIEHS, Epidemiol Branch, Mail Drop A3-05,POB 12233,111 TW Alexander Dr, Res Triangle Pk, NC 27709 USA. RI Rogan, Walter/I-6034-2012 OI Rogan, Walter/0000-0002-9302-0160 NR 44 TC 27 Z9 29 U1 2 U2 7 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0890-8567 J9 J AM ACAD CHILD PSY JI J. Am. Acad. Child Adolesc. Psychiatr. PD JUL PY 2001 VL 40 IS 7 BP 828 EP 836 DI 10.1097/00004583-200107000-00018 PG 9 WC Psychology, Developmental; Pediatrics; Psychiatry SC Psychology; Pediatrics; Psychiatry GA 445XH UT WOS:000169483500018 PM 11437022 ER PT J AU Shirani, J Lee, J Quigg, R Pick, R Bacharach, SL Dilsizian, V AF Shirani, J Lee, J Quigg, R Pick, R Bacharach, SL Dilsizian, V TI Relation of thallium uptake to morphologic features of chronic ischemic heart disease: Evidence for myocardial remodeling in noninfarcted myocardium SO JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY LA English DT Article ID LEFT-VENTRICULAR FUNCTION; CORONARY-BYPASS SURGERY; IRREVERSIBLE DEFECTS; TL-201 REINJECTION; BLOOD-FLOW; VIABILITY; CARDIOMYOPATHY; REDISTRIBUTION; REVASCULARIZATION; IMPROVEMENT AB OBJECTIVES The aim of this study was to investigate the disparity between the extent of myocardial injury as assessed by thallium and the severity of left ventricular (LV) dysfunction in chronic ischemic heart disease. BACKGROUND Although it is believed that thallium differentiates between viable and nonviable myocardium, in some patients with chronic ischemic heart disease, viable regions by thallium may fail to improve function after revascularization. METHODS Thirteen transplant candidates with chronic ischemic heart disease (LV ejection fraction = 14 +/- 6% at rest) were studied prospectively with stress-redistribution-reinjection thallium single-photon emission computed tomography. We examined pretransplantation quantitative thallium uptake and post-transplantation extent and the histological distribution of collagen replacement in infarcted and noninfarcted myocardium and in 13 age-matched control hearts. RESULTS The volume fraction of collagen varied inversely with wall thickness (r = -0.70, p < 0.001) and was higher in irreversible (30.9 +/- 15.8%) compared with reversible (20.2 +/- 12.6%, p < 0.001) or normal thallium segments (15.0 +/- 8.7%, p < 0.001). The irreversible thallium segments had lower wall thickness and more severe coronary artery narrowing (9.7 +/- 2.8 mm and 95 +/- 8%) compared with reversible (11.7 +/- 2.7 mm and 87 +/- 13%, p < 0.001) and normal thallium segments (12.8 +/- 2.6 mm and 80 +/- 14%, p < 0.001). Mean volume fraction of collagen was significantly lower in noninfarcted than it was in infarcted segments (13 +/- 6% vs. 36 +/- 13%, p < 0.001) but exceeded that in the control hearts (4 +/- 2%, p < 0.001). Noninfarcted segments had predominantly interstitial fibrosis with either microscopic or patchy areas of replacement fibrosis. CONCLUSIONS In chronic ischemic heart disease with severe LV dysfunction, patterns of normal, reversible and irreversible thallium uptake correlated with the magnitude of collagen replacement, segmental wall thickness and severity of coronary artery narrowing. The finding of scattered areas of replacement fibrosis in noninfarcted myocardium may explain the observed disparity between LV contractile dysfunction and the extent of myocardial injury assessed by thallium. (J Am Coll Cardiol 2001;38:84-90) (C) 2001 by the American College of Cardiology. C1 NHLBI, Cardiol Branch, NIH, Bethesda, MD 20892 USA. Albert Einstein Coll Med, New York, NY USA. Kyungpook Natl Univ Hosp, Taegu, South Korea. Columbia Michael Reese Hosp & Med Ctr, Chicago, IL USA. NIH, Dept Nucl Med, Bethesda, MD USA. RP Dilsizian, V (reprint author), NHLBI, Cardiol Branch, NIH, Bldg 10,Room 7B-15, Bethesda, MD 20892 USA. NR 24 TC 10 Z9 10 U1 0 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0735-1097 J9 J AM COLL CARDIOL JI J. Am. Coll. Cardiol. PD JUL PY 2001 VL 38 IS 1 BP 84 EP 90 DI 10.1016/S0735-1097(01)01320-1 PG 7 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 448KE UT WOS:000169625400013 PM 11451301 ER PT J AU Krone, RJ Hardison, RM Chaitman, BR Gibbons, RJ Sopko, G Bach, R Detre, KM AF Krone, RJ Hardison, RM Chaitman, BR Gibbons, RJ Sopko, G Bach, R Detre, KM TI Risk stratification after successful coronary revascularization: The lack of a role for routine exercise testing SO JOURNAL OF THE AMERICAN COLLEGE OF CARDIOLOGY LA English DT Article ID ARTERY DISEASE; MYOCARDIAL-INFARCTION; PROGNOSTIC VALUE; ASYMPTOMATIC RESTENOSIS; THALLIUM SCINTIGRAPHY; SPECT-THALLIUM; ANGIOPLASTY; SURGERY; TL-201; DEATH AB OBJECTIVE The objective of this study was to evaluate the American College of Cardiology/American Heart Association (ACC/AHA) guidelines fur exercise testing (EXT) after successful coronary revascularization (CR) using the Bypass Angioplasty Revascularization Investigation experience. BACKGROUND The ACC/AHA guidelines state that EXT within three years of successful CR is not useful. METHODS The 1,678 patients randomized to CR by tither angioplasty or bypass surgery were required to take symptom-limited treadmill tests one, three and five years after revascularization. RESULTS Patients who took the test at each specified time had a much lower subsequent two-year mortality than those who did not (1.9% vs. 9.4%, 3.5% vs. 12.6% and 3.3% vs. 11.0% at one, three and five years, respectively, after CR [p < 0.0001 for each]). Exercise parameters at the one- and three-year test did not improve a multivariable model of survival after including clinical parameters. Exercising to Bruce stage 3 or generating a Duke score >-6 were independently predictive of two-year survival after the five-year test. ST depression on the one-year test was associated with more revascularizations (relative risk = 1.6; p < 0.001). CONCLUSIONS Patients with stable multivessel coronary disease who rook a protocol-mandated. exercise test at one, three and five years after revascularization were at low risk for mortality in the two years subsequent to each test. Exercise parameters did not improve prediction of mortality in the two years after the one- and three-year tests. The ACC/AHA guidelines on exercise testing after CR (no value for routine testing in stable patients for three years after revascularization) are supported by these results. (J Am Coll Cardiol 2001.;38: 136-42) (C) 2001 by the American College of Cardiology. C1 Univ Pittsburgh, BARI Coordinating Ctr, Grad Sch Publ Hlth, Pittsburgh, PA 15261 USA. Washington Univ, Sch Med, St Louis, MO USA. St Louis Univ, Hlth Sci Ctr, St Louis, MO 63103 USA. Mayo Clin, Rochester, MN USA. NHLBI, Bethesda, MD 20892 USA. RP Detre, KM (reprint author), Univ Pittsburgh, BARI Coordinating Ctr, Grad Sch Publ Hlth, 127 Parran Hall,130 DeSoto St, Pittsburgh, PA 15261 USA. NR 22 TC 11 Z9 11 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0735-1097 J9 J AM COLL CARDIOL JI J. Am. Coll. Cardiol. PD JUL PY 2001 VL 38 IS 1 BP 136 EP 142 DI 10.1016/S0735-1097(01)01312-2 PG 7 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 448KE UT WOS:000169625400021 PM 11451263 ER PT J AU Dwyer, JT Evans, M Stone, EJ Feldman, HA Lytle, L Hoelscher, D Johnson, C Zive, M Yang, MH AF Dwyer, JT Evans, M Stone, EJ Feldman, HA Lytle, L Hoelscher, D Johnson, C Zive, M Yang, MH CA Catch Cooperative Res Grp TI Adolescents' eating patterns influence their nutrient intakes SO JOURNAL OF THE AMERICAN DIETETIC ASSOCIATION LA English DT Article ID PHYSICAL-ACTIVITY; CATCH; CHILDREN; SNACKS; CONSUMPTION; HEALTH; ENERGY; TRIAL; RISK C1 Tufts Univ, New England Med Ctr, Boston, MA 02111 USA. Tufts Univ, Frances Stern Nutr Ctr, Boston, MA 02111 USA. Tufts Univ, Sch Med, Boston, MA 02111 USA. Tufts Univ, Sch Nutr, Boston, MA 02111 USA. Natl Inst Alternat Med, NIH, Bethesda, MD USA. NHLBI, NIH, Bethesda, MD 20892 USA. New England Res Inst, Watertown, MA 02172 USA. Univ Minnesota, Sch Publ Hlth, Div Epidemiol, Minneapolis, MN 55455 USA. Univ Texas, Sch Publ Hlth, Houston, TX USA. Tulane Univ, Sch Publ Hlth & Trop Med, Dept Community Hlth Sci, New Orleans, LA USA. Univ Calif San Diego, Div Community Pediat, San Diego, CA 92103 USA. ABT Associates Inc, Cambridge, MA 02138 USA. RP Dwyer, JT (reprint author), Tufts Univ, New England Med Ctr, 750 Washington St,Box 783, Boston, MA 02111 USA. OI Dwyer, Johanna/0000-0002-0783-1769 FU NHLBI NIH HHS [U01-HL-39880, U01-HL-39906, U01-HL-39852] NR 30 TC 83 Z9 88 U1 4 U2 12 PU AMER DIETETIC ASSOC PI CHICAGO PA 216 W JACKSON BLVD #800, CHICAGO, IL 60606-6995 USA SN 0002-8223 J9 J AM DIET ASSOC JI J. Am. Diet. Assoc. PD JUL PY 2001 VL 101 IS 7 BP 798 EP 802 DI 10.1016/S0002-8223(01)00198-5 PG 5 WC Nutrition & Dietetics SC Nutrition & Dietetics GA 460MY UT WOS:000170312400019 PM 11478479 ER PT J AU Mori, Y Jones, M Rusk, RA Irvine, T Sahn, DJ AF Mori, Y Jones, M Rusk, RA Irvine, T Sahn, DJ TI Noninvasive assessment of left ventricular isovolumic contraction and relaxation with continuous wave Doppler aortic regurgitant velocity signals: An in vivo validation study SO JOURNAL OF THE AMERICAN SOCIETY OF ECHOCARDIOGRAPHY LA English DT Article ID IMAGED VENA-CONTRACTA; CHRONIC ANIMAL-MODEL; PRESSURE RISE; ECHOCARDIOGRAPHY; CATHETERIZATION; EFFICACY; SAFETY; AGENT AB The purpose of this study was to provide fundamental in vivo validation of a method with the use of aortic regurgitant (AR) jet signals recorded with continuous wave (CW) Doppler for assessing left ventricular (LV) isovolumic contraction and relaxation. Preliminary studies have suggested that analysis of CW Doppler AR velocity signals permits the estimation of LV positive and negative dP/dt, We studied 19 hemodynamically different states in 6 sheep with surgically induced chronic aortic regurgitation. CW AR velocity spectra and high-fidelity LV and aortic pressures were recorded simultaneously. Rates of LV pressure rise and fall (RPR and RPF) were calculated by determining the time interval between points at 1 m/s and 2.5 m/s in the deceleration and acceleration slopes of the CW Doppler AR velocity envelope (corresponding to a pressure change of 21 mm Hg). RPR and RPF calculated by CW Doppler analysis for each state were compared with the peak positive dP/dt and negative dP/dt, obtained from the corresponding high-fidelity LV pressure curve, respectively. The LV peak positive and negative dP/dt derived by catheter ranged from 817 to 2625 nun Hg/s and from 917 to 2583 mm Hg/s, respectively. Multiple regression analysis showed that Doppler RPR correlated well with catheter peak positive dP/dt (r = 0.93; mean differences, -413 +/- 250 nun Hg/s), There was also good correlation and agreement between Doppler RPF and the catheter peak negative dP/dt (r = 0.89; mean difference, -279 +/- 239 mm Hg/s), Both Doppler-determined RPR and RPF underestimated their respective LV peak dP/dt. CW Doppler AR spectra can provide a reliable noninvasive estimate of LV dP/dt and could be helpful in the serial assessment of ventricular function in patients with aortic regurgitation. C1 NHLBI, NIH, Lab Anim Med & Surg, Bethesda, MD 20892 USA. Oregon Hlth Sci Univ, Portland, OR 97201 USA. RP Jones, M (reprint author), NHLBI, NIH, Lab Anim Med & Surg, 9000 Rockville Pike,Bldg 14E,Rm 107A, Bethesda, MD 20892 USA. NR 21 TC 3 Z9 3 U1 0 U2 1 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0894-7317 J9 J AM SOC ECHOCARDIOG JI J. Am. Soc. Echocardiogr. PD JUL PY 2001 VL 14 IS 7 BP 715 EP 722 DI 10.1067/mje.2001.111477 PG 8 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 452RA UT WOS:000169871500010 PM 11447418 ER PT J AU Hunter, KA Singh, GJ Simpkins, CO AF Hunter, KA Singh, GJ Simpkins, CO TI Cyclic GMP is a measure of physiologic stress SO JOURNAL OF THE NATIONAL MEDICAL ASSOCIATION LA English DT Article DE carboxyhemoglobin; cyclic guanosine monophosphate; critical illness; carbon monoxide ID HEME OXYGENASE-1 GENE; NITRIC-OXIDE; CARBON-MONOXIDE; RAT HEPATOCYTES; ANGIOTENSIN-II; SMOOTH-MUSCLE; IN-VIVO; EXPRESSION; SYSTEM; CGMP AB This study was undertaken to determine whether patients who were critically ill evidenced elevated levels of blood cyclic guanosine monophosphate (cGMP). Cyclic guanosine monophosphate levels correlated with severity of illness as measured by the APACHE II severity of illness scoring system (p < 0.01). Cyclic guanosine monophosphate also correlated with the level of carboxyhemoglobin (HbCO) (p < 0.001). The correlation between cGMP and creatinine was p < 0.0001. Patients with end-stage disease (renal or liver) tended to have elevated levels of cGMP (p < 0.0001). We conclude that the induction of these two molecules may be linked in patients with increasing severity of illness. C1 Howard Univ, Dept Anesthesiol, NIH, Dept Surg, Bethesda, MD 20892 USA. RP Hunter, KA (reprint author), Howard Univ, Dept Anesthesiol, NIH, Dept Surg, 9000 Rockville Pike,10-2C 624, Bethesda, MD 20892 USA. NR 41 TC 0 Z9 0 U1 0 U2 0 PU NATL MED ASSOC PI WASHINGON PA 1012 10TH ST, N W, WASHINGON, DC 20001 USA SN 0027-9684 J9 J NATL MED ASSOC JI J. Natl. Med. Assoc. PD JUL-AUG PY 2001 VL 93 IS 7-8 BP 256 EP 262 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 456HK UT WOS:000170076900006 PM 11491275 ER PT J AU Hattery, D Chernomordik, V Loew, M Gannot, I Gandjbakhche, A AF Hattery, D Chernomordik, V Loew, M Gannot, I Gandjbakhche, A TI Analytical solutions for time-resolved fluorescence lifetime imaging in a turbid medium such as tissue SO JOURNAL OF THE OPTICAL SOCIETY OF AMERICA A-OPTICS IMAGE SCIENCE AND VISION LA English DT Article ID MULTIPLY SCATTERED-LIGHT; PHOTON MIGRATION; BIOLOGICAL MEDIA; SPECTROSCOPY; MODEL; CONTRAST; RESOLUTION; SIGNALS; TUMORS AB An analytical solution is developed to quantify a site-specific fluorophore lifetime perturbation that occurs, for example, when the local metabolic status is different from that of surrounding tissue. This solution may be used when fluorophores are distributed throughout a highly turbid media and the site of interest is embedded many mean scattering distances from the source and the detector. The perturbation in lifetime is differentiated from photon transit delays by random walk: theory. This analytical solution requires a priori knowledge of the tissue-scattering and absorption properties at the excitation and emission wavelengths that may be obtained from concurrent time-resolved reflection measurements. Additionally, the solution has been compared with the exact, numerically solved solution. Thus the presented solution forms the basis for practical lifetime imaging in turbid media such as tissue. (C) 2001 Optical Society of America. C1 NICHHD, Lab Integrat & Med Biophys, NIH, Bethesda, MD 20892 USA. George Washington Univ, Inst Med Imaging & Image Anal, Dept Elect & Comp Engn, Washington, DC 20052 USA. Tel Aviv Univ, Fac Engn, Dept Biomed Engn, IL-69978 Tel Aviv, Israel. RP Hattery, D (reprint author), NICHHD, Lab Integrat & Med Biophys, NIH, Bldg 12A,Room 2041, Bethesda, MD 20892 USA. NR 28 TC 30 Z9 30 U1 0 U2 0 PU OPTICAL SOC AMER PI WASHINGTON PA 2010 MASSACHUSETTS AVE NW, WASHINGTON, DC 20036 USA SN 0740-3232 J9 J OPT SOC AM A JI J. Opt. Soc. Am. A-Opt. Image Sci. Vis. PD JUL PY 2001 VL 18 IS 7 BP 1523 EP 1530 DI 10.1364/JOSAA.18.001523 PG 8 WC Optics SC Optics GA 445JD UT WOS:000169453600013 PM 11444544 ER PT J AU Vasselli, JR Yang, JC Linehan, WM White, DE Rosenberg, SA Walther, MM AF Vasselli, JR Yang, JC Linehan, WM White, DE Rosenberg, SA Walther, MM TI Lack of retroperitoneal lymphadenopathy predicts survival of patients with metastatic renal cell carcinoma SO JOURNAL OF UROLOGY LA English DT Article DE lymphatic diseases; survival; carcinoma, renal cell; neoplasm metastasis ID HIGH-DOSE INTERLEUKIN-2; PROGNOSTIC FACTORS; NEPHRECTOMY; CANCER; IMMUNOTHERAPY; THERAPY AB Purpose: Patients with metastatic renal cell carcinoma have a reported 5-year survival of 0% to 20%. The ability to predict which patients would benefit from nephrectomy and interleukin-2 (IL-2) therapy before any treatment is initiated would be useful for maximizing the advantage of therapy and improving the quality of life. Materials and Methods: A retrospective analysis of the x-rays and charts of patients treated at the National Institutes of Health Surgery Branch between 1985 and 1996, who presented with metastatic renal cancer beyond the locoregional area and the primary tumor in place, was performed. Preoperative computerized tomography or magnetic resonance imaging, or radiological reports if no scans were available, were used to obtain an estimate of the volume of retroperitoneal lymphadenopathy. Operative notes were used to evaluate whether all lymphadenopathy was resected or disease left in situ, or if any extrarenal resection, including venacavotomy, was performed. Mean survival rate was calculated from the time of nephrectomy to the time of death or last clinical followup. If patients received IL-2 therapy, the response to treatment was recorded. Mean survival and response rate for IL-2 were compared among patients in 3 separate analyses. Patients without preoperatively detected lymphadenopathy were compared with those with at least 1 cm.(3) retroperitoneal lymphadenopathy. Also, the patients who had detectable lymphadenopathy were divided into subgroups consisting of all resected, incompletely resected, unresectable and unknown if all disease was resected. Each subgroup was compared with patients without detectable preoperative lymphadenopathy. Patients with less than were compared to those with greater than 50 cm. retroperitoneal lymphadenopathy. Patients undergoing extrarenal resection at nephrectomy (complex surgery) due to direct invasion of the tumor into another intra-abdominal organ were compared with those undergoing radical nephrectomy alone, regardless of lymph node status. Statistical analysis was done with the Mantel-Cox test for comparison of survival on Kaplan-Meier curves and with Fisher's exact test for response rates for IL-2. Results: A total of 154 patients with metastatic renal cell carcinoma underwent cytoreductive nephrectomy as preparation for IL-2 based regimens. There were 82 patients with metastatic renal cell carcinoma and no preoperative retroperitoneal lymphadenopathy who survived longer (median 14.1 months) than the 72 with lymphadenopathy (median 8.5, p = 0.0004). Patients with incompletely resected, unresectable or an unknown volume resected had decreased survival compared to those with no retroperitoneal lymphadenopathy. A multivariate analysis of survival was performed evaluating the known prognostic indicators, performance status and tumor burden, as represented by the number of organs involved with metastases, and the new prognostic factor, lymphadenopathy. Lymphadenopathy was more closely associated with survival than performance status, and appeared to be a new prognostic variable. Patients with and without retroperitoneal lymphadenopathy at initial presentation had similar rates for treatment with IL-2 (54% for both groups). Of the 82 patients with no lymphadenopathy 11 (13%) had long-term survival greater than 5 years. Of the 6 complete responses to IL-2, 5 occurred in this group. Only I other patient with incompletely resected retroperitoneal lymphadenopathy survived longer than 5 years. No significant difference in survival was seen between patients who did or did not undergo complex surgery. Conclusions: Patients who presented with metastatic renal cancer and retroperitoneal lymphadenopathy had a shorter survival than those with no detectable retroperitoneal lymphadenopathy. It is warranted to continue to perform complex extrarenal resection during nephrectomy since no significant difference in the response rate for IL-2 or mean survival compared with those of patients undergoing nephrectomy alone is currently detectable. C1 NCI, Div Canc Treatment, Clin Oncol Program,Surg Branch, NIH, Bethesda, MD USA. NCI, Div Canc Treatment, Clin Oncol Program,Urol Oncol Branch, NIH, Bethesda, MD USA. RP Walther, MM (reprint author), NCI, DCS, Urol Oncol Branch, NIH, Bldg 10,Room 2B-43,10 Ctr Dr,MSC 1502, Bethesda, MD 20892 USA. NR 15 TC 88 Z9 91 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0022-5347 J9 J UROLOGY JI J. Urol. PD JUL PY 2001 VL 166 IS 1 BP 68 EP 72 DI 10.1016/S0022-5347(05)66078-8 PG 5 WC Urology & Nephrology SC Urology & Nephrology GA 442CY UT WOS:000169268700014 PM 11435825 ER PT J AU Pautler, SE McWilliams, GW Harrington, FS Walther, MM AF Pautler, SE McWilliams, GW Harrington, FS Walther, MM TI An articulating retractor holder to facilitate laparoscopic adrenalectomy and nephrectomy SO JOURNAL OF UROLOGY LA English DT Article DE kidney; laparoscopy; nephrectomy; adrenal glands; adrenalectomy ID LEARNING-CURVE; SOLO SURGERY; LIVER LOBE; EXPERIENCE; TROCAR; MODEL AB Purpose: Minimally invasive surgical approaches to renal and adrenal tumors have gained increasing use. The addition of robotic assistance and mechanical devices has decreased the number of assistants required for these often long cases. We describe an articulating arm retractor holder to aid in Liver and spleen retraction during laparoscopic surgery. Materials and Methods: The articulating retractor holder consists of 4 components, including a base rod, flexible extension arm, stainless steel precision clamp and table attachment. During laparoscopy the abdomen is visualized using standard approaches. The articulating retractor holder is clamped to the table via the base rod and brought into the surgical field using sterile technique. A retractor is placed intracorporeally to elevate the Liver or spleen and the handle is secured in place using the precision clamp. Results: The articulating retractor holder was used in all right and approximately 50% of left laparoscopic cases. Adequate visualization was obtained in all patients. There were no device failures or retractor related complications. Conclusions: The articulating retractor holder is a useful tool to aid in laparoscopic retraction that should be added to the urologist armamentarium. C1 NCI, Urol Oncol Branch, Bethesda, MD 20892 USA. NCI, Radiat Oncol Branch, Bethesda, MD 20892 USA. RP Pautler, SE (reprint author), NCI, Urol Oncol Branch, Bethesda, MD 20892 USA. NR 18 TC 4 Z9 4 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0022-5347 J9 J UROLOGY JI J. Urol. PD JUL PY 2001 VL 166 IS 1 BP 198 EP 199 DI 10.1016/S0022-5347(05)66110-1 PG 2 WC Urology & Nephrology SC Urology & Nephrology GA 442CY UT WOS:000169268700054 PM 11435857 ER PT J AU Yeo, AET Ghany, M Conry-Cantilena, C Melpolder, JC Kleiner, DE Shih, JWK Hoofnagle, JH Alter, HJ AF Yeo, AET Ghany, M Conry-Cantilena, C Melpolder, JC Kleiner, DE Shih, JWK Hoofnagle, JH Alter, HJ TI Stability of HCV-RNA level and its lack of correlation with disease severity in asymptomatic chronic hepatitis C virus carriers SO JOURNAL OF VIRAL HEPATITIS LA English DT Article DE hepatitis C virus; liver histology ID POLYMERASE CHAIN-REACTION; BLOOD MONONUCLEAR-CELLS; CHRONIC LIVER-DISEASE; LINE PROBE ASSAY; QUANTITATIVE-ANALYSIS; SERUM; QUANTIFICATION; GENOTYPES; INFECTION; TITER AB This study examines the relationship between HCV-RNA levels and disease severity in 60 individuals with chronic hepatitis C virus infection. HCV-RNA levels were quantified by the branched DNA (bDNA) assay in 445 samples (median: eight samples per patient) obtained over a median of 40.4 months (95% confidence interval (CI): 37.0-42.5). The median log HCV-RNA level was 6.77 (95% CI: 6.62-6.92) molecular equivalents/mL (MEQ/mL). The median log range of HCV-RNA levels in individual patients over the course of the study was 0.89 (95% CI: 0.69-1.16). HCV-RNA level varied over time by less than one log in 62% of patients, by 1-1.5 logs in 22% and by greater than 1.5 logs in only 17%. Univariate analysis, revealed an inverse association between HCV-RNA levels and ALT levels (P=0.037). Univariate and logistic regression analysis showed no significant association between HCV-RNA levels and either the degree of inflammation or fibrosis. In contrast, there was a significant positive association between alanine aminotransferase (ALT) levels and histological activity especially in individuals with ALTs> 100 IU/L. Hence, HCV-RNA levels: (i) almost always fell within the dynamic range of the bDNA assay; (ii) were stable in asymptomatic chronically infected patients, with only a small proportion of patients exceeding a range of 1.5 logs; (iii) did not correlate with either the extent of inflammation or degree of fibrosis. In contrast, there was a strong association between ALT level and the histological severity of liver disease. C1 NIH, Warren Grant Magnuson Clin Ctr, Dept Transfus Med, Bethesda, MD 20892 USA. NIDDKD, Liver Dis Sect, NIH, Bethesda, MD 20892 USA. NCI, Pathol Lab, NIH, Bethesda, MD 20892 USA. RP Alter, HJ (reprint author), NIH, Warren Grant Magnuson Clin Ctr, Dept Transfus Med, Bldg 10, Bethesda, MD 20892 USA. OI Kleiner, David/0000-0003-3442-4453 NR 42 TC 37 Z9 38 U1 0 U2 1 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 1352-0504 J9 J VIRAL HEPATITIS JI J. Viral Hepatitis PD JUL PY 2001 VL 8 IS 4 BP 256 EP 263 DI 10.1046/j.1365-2893.2001.00302.x PG 8 WC Gastroenterology & Hepatology; Infectious Diseases; Virology SC Gastroenterology & Hepatology; Infectious Diseases; Virology GA 456DY UT WOS:000170068900004 PM 11454176 ER PT J AU Szajner, P Weisberg, AS Wolffe, EJ Moss, B AF Szajner, P Weisberg, AS Wolffe, EJ Moss, B TI Vaccinia virus A30L protein is required for association of viral membranes with dense viroplasm to form immature virions SO JOURNAL OF VIROLOGY LA English DT Article ID MORPHOGENESIS; BIOGENESIS; PROMOTERS; STAGE; GENE; RECOMBINANT; ENVELOPES; MUTANTS; CORE AB The previously uncharacterized A30L gene of vaccinia virus has orthologs in all vertebrate poxviruses but no recognizable nonpoxvirus homologs or functional motifs. We determined that the A30L gene was regulated by a late promoter and encoded a protein of approximately 9 kDa. Immunoelectron microscopy of infected cells indicated that the A30L protein was associated with viroplasm enclosed by crescent and immature virion membranes. The A30L protein was also present in mature virions and was partially released by treatment with a nonionic detergent and reducing agent, consistent with a location in the matrix between the core and envelope, To determine the role of the A30L protein, we constructed a stringent conditional lethal mutant with an inducible A30L gene. In the absence of inducer, synthesis of viral early and late proteins occurred but the proteolytic processing of certain core proteins was inhibited, suggesting an assembly block. Inhibition of virus maturation was confirmed by electron microscopy. Under nonpermissive conditions, me observed aberrant large, dense, granular masses of viroplasm with clearly defined margins; viral crescent membranes that appeared normal except for their location at a distance from viroplasm; empty immature virions; and an absence of mature virions. The data indicated that the A30L protein is needed for vaccinia virus morphogenesis, specifically the association of the dense viroplasm with viral membranes. C1 NIAID, Viral Dis Lab, NIH, Bethesda, MD 20892 USA. George Washington Univ, Grad Program, Dept Genet, Washington, DC 20052 USA. RP Moss, B (reprint author), NIAID, Viral Dis Lab, NIH, 4 Ctr Dr,MSC 0445, Bethesda, MD 20892 USA. NR 31 TC 28 Z9 29 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD JUL PY 2001 VL 75 IS 13 BP 5752 EP 5761 DI 10.1128/JVI.75.13.5752-5761.2001 PG 10 WC Virology SC Virology GA 440KX UT WOS:000169175400004 PM 11390577 ER PT J AU Kazanji, M Tartaglia, J Franchini, G de Thoisy, B Talarmin, A Contamin, H Gessain, A de The, G AF Kazanji, M Tartaglia, J Franchini, G de Thoisy, B Talarmin, A Contamin, H Gessain, A de The, G TI Immunogenicity and protective efficacy of recombinant human T-cell leukemia/lymphoma virus type 1 NYVAC and naked DNA vaccine candidates in squirrel monkeys (Saimiri sciureus) SO JOURNAL OF VIROLOGY LA English DT Article ID HTLV-I INFECTION; LEUKEMIA-VIRUS; ENVELOPE GENE; LYMPHOCYTES-T; IMMUNE-RESPONSES; RHESUS MACAQUES; ENV GENE; GAG GENE; IMMUNIZATIONS; EXPRESSION AB We assessed the immunogenicities and efficacies of two highly attenuated vaccinia virus-derived NYVAC vaccine candidates encoding the human T-cell leukemia/lymphoma virus type 1 (HTLV-1) ear gene or both the env and gag genes in prime-boost pilot regimens in combination with naked DNA expressing the HTLV-1 envelope. Three inoculations of NYVAC HTLV-1 env at 0, 1, and 3 months followed by a single inoculation of DNA env at 9 months protected against intravenous challenge with HTLV-1-infected cells in one of three immunized squirrel monkeys. Furthermore, humoral and cell-mediated immune responses against HTLV-1 Env could be detected in this protected animal. However, priming the animal with a single dose of env DNA, followed by immunization with the NYVAC HTLV-1 gag and env vaccine at 6, 7, and 8 months, protected all three animals against challenge with HTLV-1-infected cells. With this protocol, antibodies against HTLV-1 Fm and cell-mediated responses against Env and Gag could also be detected in the protected animals. Although the relative superiority of a DNA prime-NYVAC boost regimen over addition of the Gag component as an immunogen cannot be assessed directly, our findings nevertheless show that an HTLV-1 vaccine approach is feasible and deserves further study. C1 Inst Pasteur, Lab Retrovirol, F-97306 Cayenne, French Guiana. Inst Pasteur, Ctr Primatol, F-97306 Cayenne, French Guiana. Inst Pasteur, Unite Oncol Virale, F-75724 Paris, France. Aventis Pasteur, Toronto, ON, Canada. Virogenet Corp, Troy, NY 12180 USA. NCI, Div Basic Sci, NIH, Bethesda, MD 20892 USA. RP Kazanji, M (reprint author), Inst Pasteur, Lab Retrovirol, BP 6010,23 Ave Pasteur, F-97306 Cayenne, French Guiana. NR 38 TC 25 Z9 25 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD JUL PY 2001 VL 75 IS 13 BP 5939 EP 5948 DI 10.1128/JVI.75.13.5939-5948.2001 PG 10 WC Virology SC Virology GA 440KX UT WOS:000169175400022 PM 11390595 ER PT J AU Ogert, RA Lee, MK Ross, W Buckler-White, A Martin, MA Cho, MW AF Ogert, RA Lee, MK Ross, W Buckler-White, A Martin, MA Cho, MW TI N-linked glycosylation sites adjacent to and within the V1/V2 and the V3 loops of dualtropic human immunodeficiency virus type 1 isolate DH12 gp120 affect coreceptor usage and cellular tropism SO JOURNAL OF VIROLOGY LA English DT Article ID ENVELOPE GLYCOPROTEIN; CHEMOKINE RECEPTORS; SOLUBLE CD4; HIV TYPE-1; SYNCYTIUM FORMATION; VIRAL INFECTIVITY; CELLS; NEUTRALIZATION; IDENTIFICATION; BINDING AB The envelope glycoprotein of human immunodeficiency virus type 1 (HIV-1) is extensively glycosylated, containing approximately 23 asparagine (N)-linked glycosylation sites on its gp120 subunit, In this study, specific glycosylation sites on gp120 of a dualtropic primary HIV-1 isolate, DH12, were eliminated by site-directed mutagenesis and the properties of the resulting mutant envelopes were evaluated using a recombinant vaccinia virus-based cell-to-cell fusion assay alone or in the context of viral infections. Of the glycosylation sites that were evaluated, those proximal to the V1/V2 loops (N135, N141, N156, N160) and the V3 loops (N301) of gp120 were functionally critical. The glycosylation site mutations near the V1/V2 loop compromised the use of CCR5 and CXCR4 equally. In contrast, a mutation within the V3 loop preferentially inhibited the usage of CCRS; although this mutant protein completely lost its CCR5-dependent fusion activity, it retained 50% of the wild-type fusion activity with CXCR4, The replication of a virus containing this mutation was severely compromised in peripheral blood mononuclear cells, MT-4 cells, and primary monocyte-derived macrophages. A revertant virus, which acquired second site changes in the V3 loop that resulted in an increase in net positive charge, was isolated. The revertant virus fully recovered the usage of CXCR4 but not of CCRS, thereby altering the tropism of the parental virus from dualtropic to T-tropic, These results suggest that carbohydrate moieties near the V1/V2 and the V3 loops play critical roles in maintaining proper conformation of the variable loops for optimal interaction with receptors, Our results, combined with those of previously reported studies, further demonstrate that the function of individual glycans may be virus isolate dependent. C1 NIAID, Mol Microbiol Lab, NIH, Bethesda, MD 20892 USA. RP Cho, MW (reprint author), NIAID, Mol Microbiol Lab, NIH, 9000 Rockville Pike,Bldg 4,Rm 339, Bethesda, MD 20892 USA. NR 55 TC 101 Z9 105 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD JUL PY 2001 VL 75 IS 13 BP 5998 EP 6006 DI 10.1128/JVI.75.13.5998-6006.2001 PG 9 WC Virology SC Virology GA 440KX UT WOS:000169175400028 PM 11390601 ER PT J AU Johnson, JM Nicot, C Fullen, J Ciminale, V Casareto, L Mulloy, JC Jacobson, S Franchini, G AF Johnson, JM Nicot, C Fullen, J Ciminale, V Casareto, L Mulloy, JC Jacobson, S Franchini, G TI Free major histocompatibility complex class I heavy chain is preferentially targeted for degradation by human T-cell leukemia/lymphotropic virus type 1 p12(I) protein SO JOURNAL OF VIROLOGY LA English DT Article ID TROPICAL SPASTIC PARAPARESIS; HIV-1 NEF PROTEIN; OPEN READING FRAMES; VACUOLAR H+-ATPASE; ENDOPLASMIC-RETICULUM; HTLV-I; DOWN-REGULATION; GENE-PRODUCTS; LEUKEMIA; LYMPHOCYTES AB Human T-cell leukemia virus type 1 (HTLV-1) establishes a persistent infection in the host despite a vigorous virus-specific immune response. Here we demonstrate that an HTLV-l-encoded protein, p12(I), resides in the endoplasmic reticulum (ER) and Golgi and physically binds to the free human major histocompatibility complex class I heavy chains (MHC-I-Hc) encoded by the HLA-A2, -B7, and -Cw4 alleles. As a result of this interaction, the newly synthesized MHC-I-Hc fails to associate with beta (2)-microglobulin and is retrotranslocated to the cytosol, where it is degraded by the proteasome complex. Targeting of the free MHC-I-Hc, and not the MHC-I-Hc-beta (2)-microglobulin complex, by p12(I) represents a novel mechanism of viral interference and disrupts the intracellular trafficking of MHC-I, which results in a significant decrease in surface levels of MHC-I on human T-cells. These findings suggest that the interaction of p12(I) with MHC-1-Hc may interfere with antigen presentation in vivo and facilitate escape of HTLV-1-infected cells from immune recognition. C1 NCI, Basic Res Lab, Bethesda, MD 20892 USA. NINDS, Viral Immunol Sect, Bethesda, MD 20892 USA. Univ Padua, Dept Oncol & Surg Sci, Padua, Italy. RP Franchini, G (reprint author), NCI, Basic Res Lab, 41 Lib Dr,Bldg 41,Rm D804,MSC 5055, Bethesda, MD 20892 USA. NR 56 TC 81 Z9 88 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD JUL PY 2001 VL 75 IS 13 BP 6086 EP 6094 DI 10.1128/JVI.75.13.6086-6094.2001 PG 9 WC Virology SC Virology GA 440KX UT WOS:000169175400037 PM 11390610 ER PT J AU Esser, MT Graham, DR Coren, LV Trubey, CM Bess, JW Arthur, LO Ott, DE Lifson, JD AF Esser, MT Graham, DR Coren, LV Trubey, CM Bess, JW Arthur, LO Ott, DE Lifson, JD TI Differential incorporation of CD45, CD80 (B7-1), CD86 (B7-2), and major histocompatibility complex class I and II molecules into human immunodeficiency virus type 1 virions and microvesicles: implications for viral pathogenesis and immune regulation SO JOURNAL OF VIROLOGY LA English DT Article ID HLA CLASS-II; CELLULAR PROTEINS; OPPORTUNISTIC INFECTION; ADHESION RECEPTORS; MEMBRANE-PROTEINS; LYMPHOCYTES-T; HIV; CELLS; ENVELOPE; DISEASE AB Human immunodeficiency virus (HIV) infection results in a functional impairment of CD4(+) T cells long before a quantitative decline in circulating CD4(+) T cells is evident. The mechanism(s) responsible for this functional unresponsiveness and eventual depletion of CD4(+) T cells remains unclear. Both direct effects of cytopathic infection of CD4' cells and indirect effects in which uninfected "bystander" cells are functionally compromised or killed have been implicated as contributing to the immunopathogenesis of HIV infection, Because T-cell receptor engagement of major histocompatibility complex (MHC) molecules in the absence of costimulation mediated via CD28 binding to CD80 (B7-1) or CD86 (B7-2) can Lead to anergy or apoptosis, we determined whether HIV type 1 (HIV-1) virions incorporated MHC class I (MHC-I), MHC-II, CD80, or CD86. Microvesicles produced from matched uninfected cells were also evaluated. HIV infection increased MHC-II expression on T- and B-cell lines, macrophages, and peripheral blood mononclear cells (PBMC) but did not significantly alter the expression of CD80 or CD86. HIV virions derived from all MHC-II-positive cell types incorporated high levels of MHC-II, and both virions and microvesicles preferentially incorporated CD86 compared to CD80. CD45, expressed at high levels on cells, was identified as a protein present at high levels on microvesicles but was not detected on HIV-1 virions. Virion-associated, host cell-derived molecules impacted the ability of noninfectious HIV virions to trigger death in freshly isolated PBMC, These results demonstrate the preferential incorporation or exclusion of host cell proteins by budding HIV-1 virions and suggest that host cell proteins present on HIV-1 virions may contribute to the overall pathogenesis of HIV-I infection. C1 Natl Canc Inst Frederick, SAIC Frederick, AIDS Vaccine Program, Retroviral Pathogenesis Lab, Frederick, MD 21702 USA. Natl Canc Inst Frederick, SAIC Frederick, Intramural Res Support Program, Frederick, MD 21702 USA. RP Lifson, JD (reprint author), Natl Canc Inst Frederick, SAIC Frederick, AIDS Vaccine Program, Retroviral Pathogenesis Lab, Bldg 535,5th Floor, Frederick, MD 21702 USA. RI Bess, Jr., Julian/B-5343-2012 FU NCI NIH HHS [N01-CO-560000] NR 49 TC 105 Z9 107 U1 0 U2 4 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD JUL PY 2001 VL 75 IS 13 BP 6173 EP 6182 DI 10.1128/JVI.75.13.6173-6182.2001 PG 10 WC Virology SC Virology GA 440KX UT WOS:000169175400046 PM 11390619 ER PT J AU Boyer, PL Gao, HQ Clark, PK Sarafianos, SG Arnold, E Hughes, SH AF Boyer, PL Gao, HQ Clark, PK Sarafianos, SG Arnold, E Hughes, SH TI YADD mutants of human immunodeficiency virus type 1 and Moloney murine leukemia virus reverse transcriptase are resistant to lamivudine triphosphate (3TCTP) in vitro SO JOURNAL OF VIROLOGY LA English DT Article ID STERIC HINDRANCE; DNA-SYNTHESIS; POLYMERASE; MUTAGENESIS; INHIBITORS; EXPRESSION; MECHANISM; MUTATIONS; FIDELITY; COMPLEX AB When human immunodeficiency virus type I (HIV-1) is selected for resistance to 3TC, the methionine normally present at position 184 is replaced by valine or isoleucine. Position 184 is the X of the conserved YXDD motif; positions 185 and 186 form part of the triad of aspartic acids at the polymerase active site. Structural and biochemical analysis of STC-resistant HIV-1 reverse transcriptase (RT) led to a model in which a beta -branched amino acid at position 184 would act as a steric gate. Normal deoxynucleoside triphosphates (dNTPs) could still be incorporated; the oxathiolane ring of 3TCTP would clash with the beta branch of the amino acid at position 184. This model can also explain 3TC resistance in feline immunodeficiency virus and human hepatitis B virus. However, it has been reported (14) that murine leukemia viruses (MLVs) with valine (the amino acid present in the wild type), isoleucine, alanine, serine, or methionine at the X position of the YXDD motif are all resistant to 3TC. We prepared purified wild-type MLV RT and mutant MLV RTs with methionine, isoleucine, and alanine at the X position. The behavior of these RTs aas compared to those of wild-type HIV-1 RT and of HIV-1 RT with alanine at the X position. If alanine is present at the X position, both MLV RT and HIV-1 RT are relatively resistant to 3TCTP in vitro. However, the mutant enzymes were impaired relative to their wild-type counterparts; there appears to be steric hindrance for both 3TCTP and normal dNTPs. C1 Natl Canc Inst, FCRDC, HIV Drug Resistance Program, Ft Detrick, MD 21702 USA. Natl Canc Inst, FCRDC, SAIC Frederick, Ft Detrick, MD 21702 USA. Ctr Adv Biotechnol & Med, Piscataway, NJ 08854 USA. Rutgers State Univ, Dept Chem, Piscataway, NJ 08854 USA. RP Hughes, SH (reprint author), Natl Canc Inst, FCRDC, HIV Drug Resistance Program, POB B,Bldg 539,Room 130A, Ft Detrick, MD 21702 USA. OI Sarafianos, Stefan G/0000-0002-5840-154X FU NIAID NIH HHS [AI27690, R37 AI027690] NR 29 TC 20 Z9 20 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD JUL PY 2001 VL 75 IS 14 BP 6321 EP 6328 DI 10.1128/JVI.75.14.6321-6328.2001 PG 8 WC Virology SC Virology GA 445JK UT WOS:000169454200009 PM 11413298 ER PT J AU Garcia, AD Moss, B AF Garcia, AD Moss, B TI Repression of vaccinia virus Holliday junction resolvase inhibits processing of viral DNA into unit-length genomes SO JOURNAL OF VIROLOGY LA English DT Article ID TEMPERATURE-SENSITIVE MUTANTS; CELL-FREE TRANSLATION; LINEAR MINICHROMOSOMES; POLYMERASE GENE; CONCATEMER JUNCTIONS; POXVIRUS TELOMERES; HAIRPIN TERMINI; CLEAVAGE SITES; RESOLUTION; REPLICATION AB The vaccinia virus A22R gene encodes a protein that is homologous to the bacterial enzyme RuvC and specifically cleaves and resolves four-way DNA Holliday junctions into linear duplex products. To investigate the role of the vaccinia virus Holliday junction resolvase during an infection, we constructed two recombinant viruses: vA22-HA, which has a short C-terminal epitope tag appended to the A22R open reading frame, and vA22i, in which the original A22R gene is deleted and replaced by an inducible copy. Polyacrylamide gel electrophoresis and Western blot analysis of extracts and purified virions from cells infected with vA22-HA revealed that the resolvase was expressed after the onset of DNA replication and incorporated into virion cores. vA22i exhibited a conditional replication defect. In the absence of an inducer, (i) viral protein synthesis was unaffected, (ii) late-stage viral DNA replication was reduced, (iii) most of the newly synthesized viral DNA remained in a branched or concatemeric form that caused it to be trapped at the application site during pulsed-field gel electrophoresis, (iv) cleavage of concatemer junctions was inhibited, and (v) virion morpho genesis was arrested at an immature stage. These data indicated multiple roles for the vaccinia virus Holliday junction resolvase in the replication and processing of viral DNA into unit-length genomes. C1 NIAID, Viral Dis Lab, NIH, Bethesda, MD 20892 USA. RP Moss, B (reprint author), NIAID, Viral Dis Lab, NIH, 4 Ctr Dr,MSC 0445, Bethesda, MD 20892 USA. NR 50 TC 44 Z9 46 U1 0 U2 1 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD JUL PY 2001 VL 75 IS 14 BP 6460 EP 6471 DI 10.1128/JVI.75.14.6460-6471.2001 PG 12 WC Virology SC Virology GA 445JK UT WOS:000169454200024 PM 11413313 ER PT J AU Julias, JG Ferris, AL Boyer, PL Hughes, SH AF Julias, JG Ferris, AL Boyer, PL Hughes, SH TI Replication of phenotypically mixed human immunodeficiency virus type 1 virions containing catalytically active and catalytically inactive reverse transcriptase SO JOURNAL OF VIROLOGY LA English DT Article ID MURINE LEUKEMIA-VIRUS; RNASE-H ACTIVITY; AVIAN-SARCOMA VIRUS; PLUS-STRAND DNA; RIBONUCLEASE-H; CRYSTAL-STRUCTURE; ESCHERICHIA-COLI; LACKING RIBONUCLEASE; ANGSTROM RESOLUTION; MUTATIONAL ANALYSIS AB The amount of excess polymerase and RNase H activity in human immunodeficiency virus type 1 virions was measured by using vectors that undergo a single round of replication. Vectors containing wild-type reverse transcriptase (RT), vectors encoding the D110E mutation to inactivate polymerase, and vectors encoding mutations D443A and E478Q to inactivate RNase H were constructed. 293 cells were cotransfected with different proportions of plasmids encoding these vectors to generate phenotypically mixed virions. The resulting viruses,were used to infect human osteosarcoma cells, and the relative infectivity of the viruses was determined by measuring transduction of the murine cell surface marker CD24, which is encoded by the vectors. The results indicated that there is an excess of both polymerase and RNase II activities in virions. Viral replication was reduced to 42% of wild-type levels in virions with where half of the RT molecules were predicted to be catalytically active but dropped to 3% of wild-type. levels when 25% of the RT molecules were active. However, reducing RNase H activity had a lesser effect on viral replication. As expected, based on previous work with murine leukemia virus, there was relatively inefficient virus replication when the RNase a and polymerase activities were encoded on separate vectors (D110E plus E478Q and D110E plus D443A). To determine how virus replication failed when polymerase and RNase H activities were reduced, reverse transcription intermediates were measured in vector infected cells by using quantitative real-time PCR. The results indicated that using the D110E mutation to reduce the amount of active polymerase reduced the number of reverse transcripts that were initiated and also reduced the amounts of products from the late stages of reverse transcription. If the E478Q mutation was used to reduce RNase H activity, the number of reverse transcripts that were initiated was reduced; there was also a strong effect on minus-strand transfer. C1 NCI, HIV Drug Resistance Program, Frederick, MD 21702 USA. RP Hughes, SH (reprint author), NCI, HIV Drug Resistance Program, POB B,Bldg 539,Room 130A, Frederick, MD 21702 USA. EM Hughes@ncifcrf.gov NR 69 TC 93 Z9 95 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD JUL PY 2001 VL 75 IS 14 BP 6537 EP 6546 DI 10.1128/JVI.75.14.6537-6546.2001 PG 10 WC Virology SC Virology GA 445JK UT WOS:000169454200032 PM 11413321 ER PT J AU Haddrick, M Brown, CR Plishka, R Buckler-White, A Hirsch, VM Ginsberg, H AF Haddrick, M Brown, CR Plishka, R Buckler-White, A Hirsch, VM Ginsberg, H TI Biologic studies of chimeras of highly and moderately virulent molecular clones of simian immunodeficiency virus SIVsmPBj suggest a critical role for envelope in acute AIDS virus pathogenesis SO JOURNAL OF VIROLOGY LA English DT Article ID BLOOD MONONUCLEAR-CELLS; SOLUBLE CD4 BINDING; ACUTE DISEASE; HYPERVARIABLE REGION-1; GLYCOPROTEIN GP120; IMMUNE ACTIVATION; V1/V2 REGION; TYPE-1 GP120; SIVSMMPBJ14; INFECTION AB Previous studies identified three molecular clones of the acutely pathogenic SIVsmPBj strain that varied in terms of relative in vivo pathogenicity. One clone, SIVsmPBj6.6, reproducibly induced a rapidly fatal disease in pigtailed macaques. In contrast, a highly related clone (SIVsmPBj6.9) was only minimally pathogenic in macaques. PBj6.6 and PBj6.9 shared a tyrosine substitution at position 17 in the Nef protein that is a major determinant of virulence but differed at one residue in Vpx (C89R), three residues within the envelope (D119G, R871G, G872R), and a single residue in Nef (F252L). SIVsmPBj6.9 was less efficient in inducing proliferation of resting macaque peripheral blood mononuclear cells in vitro than SIVsmPBj6.6 and exhibited a marked reduction in infectivity relative to SIVsmPBj6.6. Chimeric viruses for each of these variable residues were constructed, and their biologic properties were compared to those of the parental strains. Differences in Vpx and Nef did not alter the basic biologic phenotype of the chimeras. However, the D119G substitution in the envelope of SIVsmPBj6.9 was associated with a marked reduction in the infectivity of this virus relative to SIVsmPB6.6 An associated processing defect in gp160 of SIVsmPBj6.9 and chimeras expressing the D119G substitution suggests that a reduction in virion envelope incorporation is the mechanistic basis for reduced virion infectivity. In vivo studies revealed that substitution of the PBj6.9 amino acid into PBj6.6 (D119) abrogated the pathogenicity of this previously pathogenic virus. Introduction of the PBj6.9 G119, however, did not confer full virulence to the parental PBj6.9 virus, implicating one or all of the other four substitutions in the virulence of SIVsmPBj6.6. C1 NIAID, Mol Microbiol Lab, NIH, Rockville, MD 20852 USA. RP Hirsch, VM (reprint author), NIAID, Mol Microbiol Lab, NIH, Twinbrook Facil 2,12441 Parklawn Dr, Rockville, MD 20852 USA. NR 49 TC 8 Z9 8 U1 0 U2 0 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0022-538X J9 J VIROL JI J. Virol. PD JUL PY 2001 VL 75 IS 14 BP 6645 EP 6659 DI 10.1128/JVI.75.14.6645-6659.2001 PG 15 WC Virology SC Virology GA 445JK UT WOS:000169454200043 PM 11413332 ER PT J AU Steinel, A Parrish, CR Bloom, ME Truyen, U AF Steinel, A Parrish, CR Bloom, ME Truyen, U TI Parvovirus infections in wild carnivores SO JOURNAL OF WILDLIFE DISEASES LA English DT Review DE parvovirus; carnivores; wildlife; review article ID ALEUTIAN MINK DISEASE; FELINE PANLEUKOPENIA VIRUS; DOGS LYCAON-PICTUS; CANINE HOST-RANGE; SEROLOGIC SURVEY; MINUTE VIRUS; HEMORRHAGIC ENTERITIS; MICROBIAL PATHOGENS; CAPSID STRUCTURE; DISTEMPER VIRUS AB Various parvoviruses infect carnivores and can cause disease. In this review article the knowledge about infections of free-ranging or captive carnivores with the feline parvoviruses, feline panleukopenia virus, and canine parvovirus, including the. antigenic types CPV-2a and -2b, as well as Aleutian disease of mink virus and minute virus of canines are summarized. Particular emphasis is placed on description of the evolution of canine parvo-virus which apparent ly involved wild carnivore hosts. C1 Univ Munich, Inst Med Microbiol Infect & Epidem Dis, D-80539 Munich, Germany. Cornell Univ, New York State Coll Vet Med, James A Baker Inst Anim Hlth, Ithaca, NY 14853 USA. NIAID, Rocky Mt Labs, Persistent Viral Dis Lab, NIH, Hamilton, MT 59840 USA. RP Truyen, U (reprint author), Univ Munich, Inst Med Microbiol Infect & Epidem Dis, Veterinaerst 13, D-80539 Munich, Germany. NR 101 TC 69 Z9 73 U1 10 U2 39 PU WILDLIFE DISEASE ASSN, INC PI LAWRENCE PA 810 EAST 10TH ST, LAWRENCE, KS 66044-8897 USA SN 0090-3558 J9 J WILDLIFE DIS JI J. Wildl. Dis. PD JUL PY 2001 VL 37 IS 3 BP 594 EP 607 PG 14 WC Veterinary Sciences SC Veterinary Sciences GA 459AH UT WOS:000170227300019 PM 11504234 ER PT J AU Bluman, LG Borstelmann, NA Rimer, BK Iglehart, JD Winer, EP AF Bluman, LG Borstelmann, NA Rimer, BK Iglehart, JD Winer, EP TI Knowledge, satisfaction, and perceived cancer risk among women diagnosed with ductal carcinoma in situ SO JOURNAL OF WOMENS HEALTH & GENDER-BASED MEDICINE LA English DT Article ID QUALITY-OF-LIFE; NONINVASIVE BREAST-CANCER; INTRADUCTAL CARCINOMA; OVARIAN-CANCER; PSYCHOLOGICAL DISTRESS; IN-SITU; SUSCEPTIBILITY; EXPERIENCES; ADJUSTMENT; ADHERENCE AB The prevalence of ductal carcinoma in situ (DCIS) has increased through more widespread use of screening mammography. Little is known about what women with DCIS understand about their disease and future health. Although there is a wealth of information about the psychological characteristics of women with invasive breast cancer, there is virtually no information about women who have received treatment for DCIS. Seventy-six women diagnosed with DCIS who were identified through the Duke University Tumor Registry completed a mailed self-administered questionnaire including a broad range of items to measure knowledge, satisfaction with care, risk perceptions, and psychological distress. Women with DCIS have knowledge deficits about DCIS and breast cancer, as well as concerns about recurrence and misperceptions about the likelihood for DCIS metastasis. Women were generally satisfied with their care. They were less satisfied with information related to prognosis and with perceived support from their doctors. The results of this study suggest several areas of concern for women diagnosed with DCIS. Data about risk perceptions, knowledge, and attitudes in women diagnosed with DCIS provide important preliminary ideas for future studies. In view of the frequency of the DCIS diagnosis, future investigation should be conducted to build on these findings. C1 Dana Farber Canc Inst, Breast Oncol Ctr, Boston, MA 02115 USA. Brigham & Womens Hosp, Boston, MA 02115 USA. NCI, NIH, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. RP Winer, EP (reprint author), Dana Farber Canc Inst, Breast Oncol Ctr, 44 Binney St, Boston, MA 02115 USA. FU NCI NIH HHS [R21 CA66228] NR 40 TC 39 Z9 41 U1 0 U2 2 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1524-6094 J9 J WOMEN HEALTH GEN-B JI J. WOMENS HEALTH GENDER-BASED MED. PD JUL-AUG PY 2001 VL 10 IS 6 BP 589 EP 598 DI 10.1089/15246090152543175 PG 10 WC Public, Environmental & Occupational Health; Medicine, General & Internal; Obstetrics & Gynecology; Women's Studies SC Public, Environmental & Occupational Health; General & Internal Medicine; Obstetrics & Gynecology; Women's Studies GA 466WB UT WOS:000170668200012 PM 11559456 ER PT J AU Luft, FC Ritz, E Mann, J Radermacher, J Sharma, AM Murer, H Haller, H Schnermann, J Harrington, JT AF Luft, FC Ritz, E Mann, J Radermacher, J Sharma, AM Murer, H Haller, H Schnermann, J Harrington, JT TI Monogenic hypertension: Lessons from the genome SO KIDNEY INTERNATIONAL LA English DT Article DE aldosteronism; glucocorticoids; Liddle's syndrome; Gordon's syndrome; brachydactyly ID AUTOSOMAL-DOMINANT HYPERTENSION; EPITHELIAL SODIUM-CHANNEL; NEUROVASCULAR COMPRESSION; VENTROLATERAL MEDULLA; LIDDLES-SYNDROME; NEUROGENIC HYPERTENSION; RAT MODEL; ALDOSTERONISM; MUTATIONS; SUBUNIT C1 Humboldt Univ, Fac Med Charite, Franz Volhard Clin, Med Klin Schwerpunkt Nephrol & Hypertens, D-13125 Berlin, Germany. Humboldt Univ, Div Nephrol & Hypertens, Franz Volhard Clin, D-10098 Berlin, Germany. Univ Heidelberg, D-69115 Heidelberg, Germany. Schwabing Hosp, Munich, Germany. Univ Hannover, D-30167 Hannover, Germany. Univ Zurich, Zurich, Switzerland. NIH, Bethesda, MD 20892 USA. Tufts Univ, Sch Med, Boston, MA 02111 USA. RP Luft, FC (reprint author), Humboldt Univ, Fac Med Charite, Franz Volhard Clin, Med Klin Schwerpunkt Nephrol & Hypertens, Wiltbergstr 50, D-13125 Berlin, Germany. OI Luft, Friedrich/0000-0002-8635-1199 NR 39 TC 10 Z9 10 U1 0 U2 1 PU BLACKWELL SCIENCE INC PI MALDEN PA 350 MAIN ST, MALDEN, MA 02148 USA SN 0085-2538 J9 KIDNEY INT JI Kidney Int. PD JUL PY 2001 VL 60 IS 1 BP 381 EP 390 DI 10.1046/j.1523-1755.2001.00810.x PG 10 WC Urology & Nephrology SC Urology & Nephrology GA 446CU UT WOS:000169496000048 PM 11422778 ER PT J AU Hampshire, VA Davis, JA McNickle, CA Williams, L Eskildson, H AF Hampshire, VA Davis, JA McNickle, CA Williams, L Eskildson, H TI Retrospective comparison of rat recovery weights using inhalation and injectable anaesthetics, nutritional and fluid supplementation for right unilateral neurosurgical lesioning SO LABORATORY ANIMALS LA English DT Article DE laboratory animals; isoflurane; neurosurgical lesioning; rat AB Current veterinary standards of biomedical research support include refinements in animal models that are targeted at enhancing humane care and decreasing inter-animal variability. This ultimately results in fewer numbers of animals being used and reduction in animal experimentation through mitigation of waste as well as faster research results. 6-hydroxydopamine-lesioning of the substantia nigra using a stereotactic frame device is a common procedure and is routinely performed under pentobarbital anaesthesia with monitoring by 8 h workforces. Our programme supports the humane care and use of several protocols involving the unilateral stereotactic-lesioning of rats for the purposes of creating research models of Parkinsonianism. Such procedures are commonly performed as unilateral in order to minimize aphagia and other untoward effects of the lesion. Generally, this procedure is considered minor because it involves a small incision, a cranial burrhole, and penetration of the dura. Inflammation and/or irritation of the car canal can occur secondarily to the earbar placement procedure. Human patients undergoing similar procedures typically complain of headaches from loss of intracranial pressure; which is a transient outcome. Despite the perception of minor insult, we provided aggressive periprocedural care, and our veterinary staff documented weight loss that was often greater than 15% body weight during the first 3 days. The goal of this study was to evaluate refinements to improve this outcome. For humane concerns, and because of the need to begin experimental testing one week following surgery, a goal in the recent past has been to enhance outcome for researchers and animals by refining postoperative support modalities as well as by seeking the best anaesthetic regimen to shorten postoperative deviations from baseline. Analysis of three groups of rat patients lesioned by the same investigative group over the course of refinements made in our programme indicate that an early return of homeostasis was achieved by the use of inhalation anaesthetics as replacements for barbiturates in these models. Comparison with pentobarbital recipients also indicated that homeostasis is achieved earlier when barbiturates are utilized with fluid therapy and analgesics immediately after operative procedures rather than the next morning. C1 NINCDS, NIH, Bethesda, MD 20892 USA. RP Hampshire, VA (reprint author), Adv Vet Applicat, 7307 Nevis Rd, Bethesda, MD 20817 USA. NR 15 TC 5 Z9 5 U1 1 U2 3 PU ROYAL SOC MEDICINE PRESS LTD PI LONDON PA 1 WIMPOLE STREET, LONDON W1M 8AE, ENGLAND SN 0023-6772 J9 LAB ANIM-UK JI Lab. Anim. PD JUL PY 2001 VL 35 IS 3 BP 223 EP 229 DI 10.1258/0023677011911660 PG 7 WC Veterinary Sciences; Zoology SC Veterinary Sciences; Zoology GA 449NZ UT WOS:000169694100003 PM 11459405 ER PT J AU Metcalfe, DD Akin, C AF Metcalfe, DD Akin, C TI Mastocytosis: molecular mechanisms and clinical disease heterogeneity SO LEUKEMIA RESEARCH LA English DT Article; Proceedings Paper CT Working Conference on Mastocytosis CY AUG 31-SEP 03, 2000 CL VIENNA, AUSTRIA SP Pharmacia Updohn Diagnost GesmbH, Amgen GesmbH, Aventis Behring GesmbH, Biomedica GesmbH, Eubio, Olympus Austria GesmbH, Roche GesmbH, Wyeth Lederle Pharma GesmbH DE mastocytosis; mutation; stem cell factor ID BLOOD MONONUCLEAR-CELLS; PROTOONCOGENE C-KIT; HUMAN MAST-CELLS; BONE-MARROW; PERIPHERAL-BLOOD; SYSTEMIC MASTOCYTOSIS; URTICARIA PIGMENTOSA; ACTIVATING MUTATION; PROGENITOR CELLS; CATALYTIC DOMAIN AB Systemic mastocytosis has one unifying feature: an unexplained and pathologic increase in mast cells in specific tissues. This observation, along with clinical disease heterogeneity has long suggested that mastocytosis is a disease of complex etiology. At the same time, the last decade has witnessed significant progress in identifying the critical elements that regulate mast cell growth and development. Human mast cells are now known to arise from CD34(+) progenitors, particularly under the influence of stem cell factor (SCF). This information in turn led to the critical observation that a substantial number of patients with mastocytosis exhibit activating mutations in c-kit, the receptor for SCF. And while this observation may well be key in understanding mastocytosis, this mutation alone does not explain all heterogeneity. It now appears that other influences such as genetic polymorphisms within the host may influence the course of disease in those with KIT mutations; and that the search for additional molecular events capable of creating disease diversity must continue. Published by Elsevier Science Ltd. C1 NIAID, Lab Allerg Dis, NIH, Bethesda, MD 20892 USA. RP Metcalfe, DD (reprint author), NIAID, Lab Allerg Dis, NIH, Bldg 10,Rm 11C205,10 Ctr Dr MSC 1881, Bethesda, MD 20892 USA. NR 33 TC 63 Z9 65 U1 0 U2 2 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0145-2126 J9 LEUKEMIA RES JI Leuk. Res. PD JUL PY 2001 VL 25 IS 7 SI SI BP 577 EP 582 DI 10.1016/S0145-2126(01)00046-7 PG 6 WC Oncology; Hematology SC Oncology; Hematology GA 442AM UT WOS:000169261800010 PM 11377683 ER PT J AU Valent, P Horny, HP Escribano, L Longley, BJ Li, CY Schwartz, LB Marone, G Nunez, R Akin, C Sotlar, K Sperr, WR Wolff, K Brunning, RD Parwaresch, RM Austen, KF Lennert, K Metcalfe, DD Vardiman, JW Bennett, JM AF Valent, P Horny, HP Escribano, L Longley, BJ Li, CY Schwartz, LB Marone, G Nunez, R Akin, C Sotlar, K Sperr, WR Wolff, K Brunning, RD Parwaresch, RM Austen, KF Lennert, K Metcalfe, DD Vardiman, JW Bennett, JM TI Diagnostic criteria and classification of mastocytosis: a consensus proposal SO LEUKEMIA RESEARCH LA English DT Article; Proceedings Paper CT Working Conference on Mastocytosis CY AUG 31-SEP 03, 2000 CL VIENNA, AUSTRIA SP Pharmacia Updohn Diagnost GesmbH, Amgen GesmbH, Aventis Behring GesmbH, Biomedica GesmbH, Eubio, Olympus Austria GesmbH, Roche GesmbH, Wyeth Lederle Pharma GesmbH ID MAST-CELL DISEASE; C-KIT MUTATION; CHRONIC MYELOMONOCYTIC LEUKEMIA; BLOOD MONONUCLEAR-CELLS; ACUTE MYELOID-LEUKEMIA; HUMAN BONE-MARROW; SYSTEMIC MASTOCYTOSIS; URTICARIA PIGMENTOSA; PERIPHERAL-BLOOD; GENERALIZED MASTOCYTOSIS AB The term 'mastocytosis' denotes a heterogeneous group of disorders characterized by abnormal growth and accumulation of mast cells (MC) in one or more organ systems. Over the last 20 years, there has been an evolution in accepted classification systems for this disease. In light of such developments and novel useful markers, it seems appropriate now to re-evaluate and update the classification of mastocytosis. Here, we propose criteria to delineate categories of mastocytosis together with an updated consensus classification system. In this proposal, the diagnosis cutaneous mastocytosis (CM) is based on typical clinical and histological skin lesions and absence of definitive signs (criteria) of systemic involvement. Most patients with CM are children and present with maculopapular cutaneous mastocytosis (= urticaria pigmentosa, UP). Other less frequent forms of CM are diffuse cutaneous mastocytosis (DCM) and mastocytoma of skin. Systemic mastocytosis (SM) is commonly seen in adults and defined by multifocal histological lesions in the bone marrow (affected almost invariably) or other extracutaneous organs (major criteria) together with cytological and biochemical signs (minor criteria) of systemic disease (SM-criteria). SM is further divided into the following categories: indolent systemic mastocytosis (ISM), SM with an associated clonal hematologic non-mast cell lineage disease (AKNMD), aggressive systemic mastocytosis (ASM), and mast cell leukemia (MCL). Patients with ISM usually have maculopapular skin lesions and a good prognosis. In the group with associated hematologic disease, the AHNMD should be classified according to FAB/WHO criteria. ASM is characterized by impaired organ-function due to infiltration of the bone marrow, liver, spleen, GI-tract, or skeletal system, by pathologic MC. MCL is a 'high-grade' leukemic disease defined by increased numbers of MC in bone marrow smears (greater than or equal to 20%) and peripheral blood, absence of skin lesions, multiorgan failure, and a short survival. In typical cases, circulating MC amount to greater than or equal to 10% of leukocytes (classical form of MCL). Mast cell sarcoma is a unifocal tumor that consists of atypical MC and shows a destructive growth without (primary) systemic involvement. This high-grade malignant MC disease has to be distinguished from a localized benign mastocytoma in either extracutaneous organs (= extracutaneous mastocytoma) or skin. Depending on the clinical course of mastocytosis and development of an AHNMD, patients can shift from one category of MC disease into another. In all categories, mediator-related symptoms may occur and may represent a serious clinical problem. All categories of mastocytosis should be distinctively separated from reactive MC hyperplasia, MC activation syndromes, and a more or less pronounced increase in MC in myelogenous malignancies other than mastocytosis. Criteria proposed in this article should be helpful in this regard. (C) 2001 Elsevier Science Ltd. All rights reserved. C1 Univ Vienna, Div Hematol, Dept Internal Med 1, A-1090 Vienna, Austria. Med Univ Lubeck, Inst Pathol, D-23538 Lubeck, Germany. Hosp Ramon y Cajal, Unidad Mastocitosis, Serv Hematol, E-28034 Madrid, Spain. Columbia Univ, Dept Dermatol, New York, NY 10027 USA. Columbia Univ, Dept Pathol, New York, NY 10027 USA. Mayo Clin & Mayo Fdn, Div Hematopathol, Rochester, MN 55905 USA. Virginia Commonwealth Univ, Med Coll Virginia, Dept Internal Med, Div Rheumatol Allergy & Immunol, Richmond, VA 23298 USA. Univ Naples Federico II, Cattedra Immunol Clin & Allergol, Fac Med & Chirurg, Naples, Italy. NIAID, Lab Allerg Dis, NIH, Bethesda, MD 20892 USA. Univ Tubingen, Inst Pathol, D-7400 Tubingen, Germany. Univ Vienna, Dept Dermatol, A-1090 Vienna, Austria. Univ Minnesota, Dept Lab Med & Pathol, Minneapolis, MN 55455 USA. Univ Kiel, Inst Hematopathol & Lymph Node Registry, Kiel, Germany. Harvard Univ, Brigham & Womens Hosp, Sch Med, Div Rheumatol Allergy & Immunol, Boston, MA 02115 USA. Univ Chicago, Dept Pathol, Hematopathol Sect, Chicago, IL 60637 USA. Univ Rochester, Med Ctr, Ctr Canc, Med Oncol Unit, Rochester, NY 14642 USA. RP Valent, P (reprint author), Univ Vienna, Div Hematol, Dept Internal Med 1, Waehringer Guertel 18-20, A-1090 Vienna, Austria. NR 163 TC 600 Z9 621 U1 2 U2 9 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0145-2126 J9 LEUKEMIA RES JI Leuk. Res. PD JUL PY 2001 VL 25 IS 7 SI SI BP 603 EP 625 DI 10.1016/S0145-2126(01)00038-8 PG 23 WC Oncology; Hematology SC Oncology; Hematology GA 442AM UT WOS:000169261800013 PM 11377686 ER PT J AU Akin, C Scott, LM Metcalfe, DD AF Akin, C Scott, LM Metcalfe, DD TI Slowly progressive systemic mastocytosis with high mast-cell burden and no evidence of a non-mast-cell hematologic disorder: an example of a smoldering case? SO LEUKEMIA RESEARCH LA English DT Article; Proceedings Paper CT Working Conference on Mastocytosis CY AUG 31-SEP 03, 2000 CL VIENNA, AUSTRIA SP Pharmacia Updohn Diagnost GesmbH, Amgen GesmbH, Aventis Behring GesmbH, Biomedica GesmbH, Eubio, Olympus Austria GesmbH, Roche GesmbH, Wyeth Lederle Pharma GesmbH DE bone marrow; classification; mastocytosis; urticaria pigmentosa ID BLOOD MONONUCLEAR-CELLS; C-KIT; MUTATION; DISEASE AB A 43-year-old man with extensive systemic mastocytosis with poor prognostic indicators but no overt hematologic abnormality is described. This patient's clinical presentation and course are consistent with the newly proposed 'smoldering mastocytosis' category. Long-term follow-up of patients is needed to determine whether they may be at higher risk for progression into more aggressive categories. Published by Elsevier Science Ltd. C1 NIAID, Lab Allerg Dis, NIH, Bethesda, MD 20892 USA. RP Akin, C (reprint author), NIAID, Lab Allerg Dis, NIH, Room 11C205,10 Ctr Dr,MSC-1881, Bethesda, MD 20892 USA. NR 8 TC 23 Z9 25 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0145-2126 J9 LEUKEMIA RES JI Leuk. Res. PD JUL PY 2001 VL 25 IS 7 SI SI BP 635 EP 638 DI 10.1016/S0145-2126(01)00023-6 PG 4 WC Oncology; Hematology SC Oncology; Hematology GA 442AM UT WOS:000169261800015 PM 11377688 ER PT J AU Fishbein, KW McConville, P Spencer, RGS AF Fishbein, KW McConville, P Spencer, RGS TI The lever-coil: a simple, inexpensive sensor for respiratory and cardiac motion in MRI experiments SO MAGNETIC RESONANCE IMAGING LA English DT Article DE motion artifacts; respiratory gating; cardiac triggering; magnetic resonance imaging; motion detection ID IN-VIVO; DEVICE; ANIMALS; HEART; NMR AB Thoracic and abdominal magnetic resonance imaging studies generally require some type of compensation for respiratory and cardiac motions in order to yield artifact-free images with good signal-to-noise ratio. Most techniques for respiratory compensation require the use of a non-NMR sensing device to monitor the subject's chest motion, while cardiac motion compensation generally requires the use of ECG leads within the magnet. An inductive pickup coil placed on the subject's chest is perhaps the simplest and least expensive means of monitoring respiration in a MR scanner. However, due to inductive coupling between the pickup coil, radio frequency resonator and gradient set, this arrangement often results in both NMR and respiratory signal artifacts and can also present a burn hazard to the subject depending on the placement and orientation of the pickup coil. Moreover, the presence of a pickup coil on the chest can degrade local magnetic field homogeneity and thus degrade image quality. Similar problems arise when ECG leads must be connected to the subject for cardiac monitoring and gating. To preserve the benefits of the simple pickup coil while circumventing these limitations, a "lever-coil" sensor is presented in which a pickup coil is mechanically coupled to the subject but is not located within the resonator or gradient coil. This results in much lower mutual inductance between the pickup coil and the resonator or gradients. The optimization of the geometry of the apparatus is discussed and lever-coil signal traces are shown which demonstrate the sensor's ability to simultaneously detect both respiratory and cardiac motion in mice. Finally. respiratory-gated and cardiac-triggered spin echo images of the rat abdomen and mouse heart are presented to demonstrate the utility of the lever-coil sensor. (C) 2001 Elsevier Science Inc. All rights reserved. C1 NIA, Nucl Magnet Resonance Unit, NIH, Baltimore, MD 21224 USA. RP Spencer, RGS (reprint author), NIA, Nucl Magnet Resonance Unit, NIH, Baltimore, MD 21224 USA. OI Fishbein, Kenneth/0000-0002-6353-4603 NR 21 TC 18 Z9 19 U1 1 U2 2 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0730-725X J9 MAGN RESON IMAGING JI Magn. Reson. Imaging PD JUL PY 2001 VL 19 IS 6 BP 881 EP 889 DI 10.1016/S0730-725X(01)00403-9 PG 9 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 473PH UT WOS:000171054200016 PM 11551730 ER PT J AU Duyn, JH Tan, CX van Gelderen, P Yongbi, MN AF Duyn, JH Tan, CX van Gelderen, P Yongbi, MN TI High-sensitivity single-shot perfusion-weighted fMRI SO MAGNETIC RESONANCE IN MEDICINE LA English DT Article DE MRI; perfusion imaging; CBF; arterial spin-labeling; fMRI ID STIMULATION; SIGNAL AB A method is presented for measurement of perfusion changes during brain activation using a single-shot pulsed spin labeling technique. By employing a double-inversion labeling strategy, stationary tissue (background) signal was suppressed while minimally affecting perfusion sensitivity. This allowed omission of the otherwise required reference scan, resulting in twofold-improved temporal resolution. The method was applied to visual and motor cortex activation studies in humans, and compared to standard FAIR-type perfusion labeling techniques. Experiments performed at 1.5T and 3.0T indicate a close to 90% suppression of background signal, at a cost of an 11% and 9%, respectively reduction in perfusion signal. Combined with the twofold increase in signal averaging, and a reduction in background signal fluctuations, this resulted in a 64% (1.5T, N = 3) and a 128% (3T, N = 4) overall improvement in sensitivity for the detection of activation-related perfusion changes. Published 2001 Wiley-Liss, Inc.(dagger) C1 NINDS, Lab Diagnost Radiol Res, NIH, Bethesda, MD 20892 USA. NINDS, Lab Funct & Mol Imaging, NIH, Bethesda, MD 20892 USA. RP Duyn, JH (reprint author), NINDS, Lab Diagnost Radiol Res, NIH, Bldg 10,Room B1D-108,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Duyn, Jozef/F-2483-2010 NR 12 TC 24 Z9 24 U1 0 U2 0 PU JOHN WILEY & SONS INC PI NEW YORK PA 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0740-3194 J9 MAGNET RESON MED JI Magn. Reson. Med. PD JUL PY 2001 VL 46 IS 1 BP 88 EP 94 DI 10.1002/mrm.1163 PG 7 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 447FN UT WOS:000169561000012 PM 11443714 ER PT J AU Cohen, RM Kang, A Gulick, C AF Cohen, RM Kang, A Gulick, C TI Quantitative trait loci affecting the behavior of A/J and CBA/J intercross mice in the elevated plus maze SO MAMMALIAN GENOME LA English DT Article ID PHOTORECEPTOR CGMP PHOSPHODIESTERASE; RETINAL DEGENERATION; MENDELIAN FACTORS; BETA-SUBUNIT; MOUSE; ANXIETY; GENE; RAT; NEURONS; MODELS AB How allelic diversity affects neural mechanisms to pro duce behavioral variation is largely unknown. The elevated plus maze, consisting of open and closed arms, has been used as a model of behavioral variation in rodent exploration. Under dim illumination the nature of the sensory stimuli that influence arm choice is uncertain. Two inbred mouse strains, A/J (Tyr(c)/Tyr(c) the albino phenotype, mutation in tyrosinase) with a strong preference for closed arm entry, and CBA/J (Pdeb(rdl)/Pdeb(rdl) the retinal degeneration phenotype, mutation in the P-subunit of rod cGMP phosphodiesterase), with a weak preference for open arm entry, were studied under varying light. Because behavioral differences persist under red light, variation in light perception is not likely to fully account for variation in arm choice. To identify genetic factors influencing arm choice (100 x Open arm entries/Total arm entries) quantitative trait loci analyses (QTL) were performed on (A/J x CBA/J)F-2 mice. Two QTLs, one of which includes PDEB, were identified on Chr 5 (LOD > 10) and account for > 30% of the behavioral variation in arm preference. Tyr (Chr 7, 44 cM) was linked to closed arm entries but not arm preference, and is unlikely to be acting through a direct effect on light perception, because A/J arm entries were not affected by red light and there was no interaction with PDEB in the (A/J x CBA/J)F-2 mice. Whether the candidate QTLs on Chr 5 affect arm choice through an effect on light perception is unknown, but phenotypic differences between F-2 mice with retinal degeneration and CBA/J mice and F-2 mice with albinism and A/J mice suggest that factors other than light sensitivity contribute to arm preference in these two strains. C1 NIMH, Cerebral Metab Lab, NIH, Bethesda, MD 20892 USA. NIMH, Geriatr Psychiat Branch, NIH, Bethesda, MD 20892 USA. RP Cohen, RM (reprint author), NIMH, Cerebral Metab Lab, NIH, Bldg 10,Room 3n218,10 Ctr Dr,MSC 1274, Bethesda, MD 20892 USA. NR 37 TC 21 Z9 22 U1 0 U2 2 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0938-8990 J9 MAMM GENOME JI Mamm. Genome PD JUL PY 2001 VL 12 IS 7 BP 501 EP 507 DI 10.1007/s00335-001-2047-5 PG 7 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 448KY UT WOS:000169627100003 PM 11420611 ER PT J AU Summers, TJ Thomas, JW Lee-Lin, SQ Maduro, VVB Idol, JR Green, ED AF Summers, TJ Thomas, JW Lee-Lin, SQ Maduro, VVB Idol, JR Green, ED TI Comparative physical mapping of targeted regions of the rat genome SO MAMMALIAN GENOME LA English DT Article ID INTEGRATED GENETIC-LINKAGE; RADIATION HYBRID MAP; LABORATORY RAT; SEQUENCE-ANALYSIS; HUMAN-CHROMOSOMES; HYBRIDIZATION; CLONES; HUMAN-CHROMOSOME-7; MARKERS; KB AB The comparative mapping and sequencing of vertebrate genomes is now a key priority for the Human Genome Project. In addition to finishing the human genome sequence and generating a 'working draft' of the mouse genome sequence, significant attention is rapidly turning to the analysis of other model organisms, such as the laboratory rat (Rattus norvegicus). As a complement to genome-wide mapping and sequencing efforts, it is often important to generate detailed maps and sequence data for specific regions of interest. Using an adaptation of our previously described approach for constructing mouse comparative and physical maps, we have established a general strategy for targeted mapping of the rat genome. Specifically, we constructed a framework comparative map of human Chromosome (Chr) 7 and the orthologous regions of the rat genome, as well as two large (>1-Mb) P1-derived artificial chromosome (PAC)-based physical maps. Generation of these physical maps involved the use of mouse-derived probes that cross-hybridized with rat PAC clones. The first PAC map encompasses the cystic fibrosis transmembrane conductance regulator gene (Cftr), while the second map allows a three-species comparison of a genomic region containing intra- and inter-chromosomal evolutionary rearrangements. The studies reported here further demonstrate that cross-species hybridization between related animals, such as rat and mouse, can be readily used for the targated construction of clone-based physical maps, thereby accelerating the analysis of biologically interesting regions of vertebrate genomes. C1 NHGRI, Genome Technol Branch, NIH, Bethesda, MD 20892 USA. RP Green, ED (reprint author), NHGRI, Genome Technol Branch, NIH, 49 Convent Dr,Bldg 49,Rm 2A08, Bethesda, MD 20892 USA. NR 29 TC 12 Z9 12 U1 0 U2 0 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 0938-8990 J9 MAMM GENOME JI Mamm. Genome PD JUL PY 2001 VL 12 IS 7 BP 508 EP 512 DI 10.1007/s003350020021 PG 5 WC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 448KY UT WOS:000169627100004 PM 11420612 ER PT J AU Kenworthy, AK AF Kenworthy, AK TI Imaging protein-protein interactions using fluorescence resonance energy transfer microscopy SO METHODS LA English DT Article ID GPI-ANCHORED PROTEINS; CELL-SURFACE; LIVING CELLS; PHOTOBLEACHING KINETICS; FRET MICROSCOPY; APICAL SURFACE; CHOLERA-TOXIN; 2 DIMENSIONS; CYCLIC-AMP; MDCK CELLS AB Fluorescence resonance energy transfer (FRFT) detects the proximity of fluorescently labeled molecules over distances > 100 Angstrom. When performed in a fluorescence microscope, FRET can be used to map protein-protein interactions in vivo. We here describe a FRET microscopy method that can be used to determine whether proteins that are colocalized at the level of light microscopy interact with one another. This method can be implemented using digital microscopy systems such as a confocal microscope or a wide-field fluorescence microscope coupled to a charge-coupled device (CCD) camera. It is readily applied to samples prepared with standard immunofluorescence techniques using antibodies labeled with fluorescent dyes that act as a donor and acceptor pair for FRET. Energy transfer efficiencies are quantified based on the release of quenching of donor fluorescence due to FRET, measured by comparing the intensity of donor fluorescence before and after complete photobleaching of the acceptor. As described, this method uses Cy3 and Cy5 as the donor and acceptor fluorophores, but can be adapted for other FRET pairs Including cyan fluorescent protein and yellow fluorescent protein, (C) 2001 Academic Press. C1 Johns Hopkins Univ, Dept Biol, Baltimore, MD 21218 USA. RP Kenworthy, AK (reprint author), NICHD, Cell Biol & Metab Branch, NIH, Bldg 18T,Room 101, Bethesda, MD 20892 USA. EM kenworta@mail.nih.gov FU NIDDK NIH HHS [5PO1DK-44375] NR 56 TC 356 Z9 361 U1 14 U2 119 PU ACADEMIC PRESS INC ELSEVIER SCIENCE PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1046-2023 EI 1095-9130 J9 METHODS JI Methods PD JUL PY 2001 VL 24 IS 3 BP 289 EP 296 DI 10.1006/meth.2001.1189 PG 8 WC Biochemical Research Methods; Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 447MJ UT WOS:000169576900010 PM 11403577 ER PT J AU Richter, BWM Mir, SS Eiben, LJ Lewis, J Reffey, SB Frattini, A Tian, L Frank, S Youle, RJ Nelson, DL Notarangelo, LD Vezzoni, P Fearnhead, HO Duckett, CS AF Richter, BWM Mir, SS Eiben, LJ Lewis, J Reffey, SB Frattini, A Tian, L Frank, S Youle, RJ Nelson, DL Notarangelo, LD Vezzoni, P Fearnhead, HO Duckett, CS TI Molecular cloning of ILP-2, a novel member of the inhibitor of apoptosis protein family SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Article ID ONCOGENE-DEPENDENT APOPTOSIS; CYTOCHROME-C; CASPASE ACTIVATION; NEGATIVE REGULATION; APAF-1 APOPTOSOME; IAP GENES; KAPPA-B; XIAP; BCL-X(L); COMPLEX AB Inhibitor of apoptosis protein (IAP)-like protein-1 (ILP-1) (also known as X-linked IAP [XIAP] and mammalian IAP homolog A [MIHA]) is a potent inhibitor of apoptosis and exerts its effects, at least in part, by the direct association with and inhibition of specific caspases. Here, we describe the molecular cloning and characterization of a human gene related to ILP-1, termed ILP-2. Despite high homology to ILP-1, ILP-2 is encoded by a distinct gene, which in normal tissues is expressed solely in testis. In contrast to ILP-1, overexpression of ILP-2 had no protective effect on apoptosis mediated by Fas (also known as: CD95) or tumor necrosis factor. However, ILP-2 potently inhibited apoptosis induced by overexpression of Bax or by coexpression of caspase 9 with Apaf-1, and preincubation of cytosolic extracts with ILP-2 abrogated caspase activation in vitro. A processed form of caspase 9 could be coprecipitated with ILP-2 from cells, suggesting a physical interaction between ILP-2 and caspase 9. Thus, ILP-2 is a novel IAP family member with restricted specificity for caspase 9. C1 NCI, Metab Branch, Div Clin Sci, NIH, Bethesda, MD 20892 USA. NINDS, Surg Neurol Branch, NIH, Bethesda, MD 20892 USA. NCI, Frederick Canc Res & Dev Ctr, NIH, Frederick, MD USA. CNR, Ist Tecnol Biomed Avanzate, I-20131 Milan, Italy. Univ Brescia, Dept Pediat, Brescia, Italy. RP Duckett, CS (reprint author), NCI, Metab Branch, Div Clin Sci, NIH, 10 Ctr Dr,Room 6B-05, Bethesda, MD 20892 USA. RI Fearnhead, Howard/D-4826-2012; Notarangelo, Luigi/F-9718-2016; OI Notarangelo, Luigi/0000-0002-8335-0262; Fearnhead, Howard/0000-0002-8054-9794 NR 48 TC 64 Z9 74 U1 0 U2 2 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0270-7306 J9 MOL CELL BIOL JI Mol. Cell. Biol. PD JUL PY 2001 VL 21 IS 13 BP 4292 EP 4301 DI 10.1128/MCB.21.13.4292-4301.2001 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 441MD UT WOS:000169233400020 PM 11390657 ER PT J AU Natarajan, K Meyer, MR Jackson, BM Slade, D Roberts, C Hinnebusch, AG Marton, MJ AF Natarajan, K Meyer, MR Jackson, BM Slade, D Roberts, C Hinnebusch, AG Marton, MJ TI Transcriptional profiling shows that Gcn4p is a master regulator of gene expression during amino acid starvation in yeast SO MOLECULAR AND CELLULAR BIOLOGY LA English DT Review ID PROTEIN-KINASE GCN2; SACCHAROMYCES-CEREVISIAE ENCODES; TRANSFER-RNA; BIOSYNTHETIC-PATHWAY; PEROXISOME PROLIFERATION; TRANSLATIONAL ACTIVATION; ASPERGILLUS-NIDULANS; NUTRIENT LIMITATION; HIS4 TRANSCRIPTION; MOLECULAR ANALYSIS AB Starvation for amino acids induces Gcn4p, a transcriptional activator of amino acid biosynthetic genes in Saccharomyces cerevisiae. In an effort to identify all genes regulated by Gcn4p during amino acid starvation, we performed cDNA microarray analysis. Data from 21 pairs of hybridization experiments using two different strains derived from S288c revealed that more than 1,000 genes were induced, and a similar number were repressed, by a factor of 2 or more in response to histidine starvation imposed by 3-aminotriazole (3AT). Profiling of a gcn4 Delta strain and a constitutively induced mutant showed that Gcn4p is required for the full induction by 3AT of at least 539 genes, termed Gcn4p targets. Genes in every amino acid biosynthetic pathway except cysteine and genes encoding amino acid precursors, vitamin biosynthetic enzymes, peroxisomal components, mitochondrial carrier proteins, and autophagy proteins were all identified as Gcn4p targets. Unexpectedly, genes involved in amino acid biosynthesis represent only a quarter of the Gcn4p target genes. Gcn4p also activates genes involved in glycogen homeostasis, and mutant analysis showed that Gcn4p suppresses glycogen levels in amino acid-starved cells. Numerous genes encoding protein kinases and transcription factors were identified as targets, suggesting that Gcn4p is a master regulator of gene expression. Interestingly, expression profiles for 3AT and the alkylating agent methyl methanesulfonate (MMS) overlapped extensively, and MMS induced GCN4 translation. Thus, the broad transcriptional response evoked by Gcn4p is produced by diverse stress conditions. Finally, profiling of a gcn4 Delta mutant uncovered an alternative induction pathway operating at many Gcn4p target genes in histidine-starved cells. C1 NICHHD, Lab Gene Regulat & Dev, Bethesda, MD 20892 USA. Rosetta Inpharmat, Kirkland, WA 98034 USA. RP Hinnebusch, AG (reprint author), NICHHD, Lab Gene Regulat & Dev, Bldg A,Rm B1A13, Bethesda, MD 20892 USA. NR 111 TC 433 Z9 449 U1 2 U2 32 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0270-7306 J9 MOL CELL BIOL JI Mol. Cell. Biol. PD JUL PY 2001 VL 21 IS 13 BP 4347 EP 4368 DI 10.1128/MCB.21.13.4347-4368.2001 PG 22 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 441MD UT WOS:000169233400026 PM 11390663 ER PT J AU Kee, SH Steinert, PM AF Kee, SH Steinert, PM TI Microtubule disruption in keratinocytes induces cell-cell adhesion through activation of endogenous E-cadherin SO MOLECULAR BIOLOGY OF THE CELL LA English DT Article ID MUSCARINIC ACETYLCHOLINE-RECEPTORS; HUMAN EPIDERMAL-KERATINOCYTES; SKELETAL-MUSCLE CELLS; SIGNAL-TRANSDUCTION; CATENIN COMPLEX; POSTTRANSLATIONAL MODIFICATION; DETYROSINATED MICROTUBULES; TYROSINE PHOSPHORYLATION; FEEDBACK INTERACTIONS; EPITHELIAL-CELLS AB The association of the cytoskeleton with the cadherin-catenin complex is essential for strong cell-cell adhesion in epithelial cells. In this study, we have investigated the effect of microtubule organization on cell-cell adhesion in differentiating keratinocytes. When microtubules of normal human epidermal keratinocytes (NHEKs) grown in low calcium media (0.05 mM) were disrupted with nocodazole or colcemid, cell-cell adhesion was induced through relocalization of the E-cadherin-catenin-actin complex to the cell periphery. This was accompanied by actin polymerization. Also, it was found that microtubule disruption-induced cell-cell adhesion was significantly reduced in more advanced differentiated keratinocytes. For example, when NHEK cells cultured under high calcium (1.2 mM) for 8 d and then in low calcium for I d were treated with nocodazole, there was no induction of cell-cell adhesion. Also long-term treatment of a phorbol ester for 48 h inhibited nocodazole-induced cell-cell adhesion of NHEK. Furthermore, this nocodazole-induced cell-cell adhesion could be observed in squamous cancer cell lines (A431 and SCC-5, -9, and -25) under low calcium condition, but not in the keratinocyte cell lines derived from normal epidermis (HaCaT, RHEK). On the other hand, HaCaT cells continuously cultivated in low calcium media regained a less differentiated phenotype such as decreased expression of cytokeratin 10, and increased K5; these changes were accompanied with inducibility of cell-cell adhesion by nocodazole. Together, our results suggest that microtubule disruption can induce the cell-cell adhesion via activation of endogenous E-cadherin in non- or early differentiating keratinocytes. However, this is no longer possible in advanced terminally differentiating keratinocytes, possibly due to irreversible changes effected by cell envelope barrier formation. C1 NIAMSD, Skin Biol Lab, NIH, Bethesda, MD 20892 USA. RP Steinert, PM (reprint author), NIAMSD, Skin Biol Lab, NIH, Bethesda, MD 20892 USA. NR 50 TC 23 Z9 26 U1 0 U2 0 PU AMER SOC CELL BIOLOGY PI BETHESDA PA 8120 WOODMONT AVE, STE 750, BETHESDA, MD 20814-2755 USA SN 1059-1524 J9 MOL BIOL CELL JI Mol. Biol. Cell PD JUL PY 2001 VL 12 IS 7 BP 1983 EP 1993 PG 11 WC Cell Biology SC Cell Biology GA 461DU UT WOS:000170350300006 PM 11451997 ER PT J AU Huizing, M Sarangarajan, R Strovel, E Zhao, Y Gahl, WA Boissy, RE AF Huizing, M Sarangarajan, R Strovel, E Zhao, Y Gahl, WA Boissy, RE TI AP-3 mediates tyrosinase but not TRP-1 trafficking in human melanocytes SO MOLECULAR BIOLOGY OF THE CELL LA English DT Article ID HERMANSKY-PUDLAK-SYNDROME; PIGMENT GRANULE BIOGENESIS; PROTEIN COMPLEX; ADAPTER COMPLEX; CYTOPLASMIC TAIL; LATE ENDOSOMES; OCULOCUTANEOUS ALBINISM; LOCUS HETEROGENEITY; VESICLE FORMATION; SORTING SIGNALS AB Patients with Hermansky-Pudlak syndrome type 2 (HPS-2) have mutations in the beta 3A subunit of adaptor complex-3 (AP-3) and functional deficiency of this complex. AP-3 serves as a coat protein in the formation of new vesicles, including, apparently, the platelet's dense body and the melanocyte's melanosome. We used HPS-2 melanocytes in culture to determine the role of AP-3 in the trafficking of the melanogenic proteins tyrosinase and tyrosinase-related protein-1 (TRP-1). TRP-1 displayed a typical melanosomal pattern in both normal and HPS-2 melanocytes. In contrast, tyrosinase exhibited a melanosomal (i.e., perinuclear and dendritic) pattern in normal cells but only a perinuclear pattern in the HPS-2 melanocytes. In addition, tyrosinase exhibited a normal pattern of expression in HPS-2 melanocytes transfected with a cDNA encoding the beta 3A subunit of the AP-3 complex. This suggests a role for AP-3 in the normal trafficking of tyrosinase to premelanosomes, consistent with the presence of a dileucine recognition signal in the C-terminal portion of the tyrosinase molecule. In the AP-3-deficient cells, tyrosinase was also present in structures resembling late endosomes or multivesicular bodies; these vesicles contained exvaginations devoid of tyrosinase. This suggests that, under normal circumstances, AP-3 may act on multivesicular bodies to form tyrosinase-containing vesicles destined to fuse with premelanosomes. Finally, our studies demonstrate that tyrosinase and TRP-1 use different mechanisms to reach their premelanosomal destination. C1 Univ Cincinnati, Dept Dermatol, Cincinnati, OH 45267 USA. NICHHD, Sect Human Biochem Genet, Heritable Disorders Branch, NIH, Bethesda, MD 20892 USA. RP Boissy, RE (reprint author), Univ Cincinnati, Dept Dermatol, Cincinnati, OH 45267 USA. FU NIAMS NIH HHS [AR-45429, R01 AR045429] NR 58 TC 103 Z9 108 U1 0 U2 0 PU AMER SOC CELL BIOLOGY PI BETHESDA PA 8120 WOODMONT AVE, STE 750, BETHESDA, MD 20814-2755 USA SN 1059-1524 J9 MOL BIOL CELL JI Mol. Biol. Cell PD JUL PY 2001 VL 12 IS 7 BP 2075 EP 2085 PG 11 WC Cell Biology SC Cell Biology GA 461DU UT WOS:000170350300013 PM 11452004 ER PT J AU Ohmori, H Friedberg, EC Fuchs, RPP Goodman, MF Hanaoka, F Hinkle, D Kunkel, TA Lawrence, CW Livneh, Z Nohmi, T Prakash, L Prakash, S Todo, T Walker, GC Wang, ZG Woodgate, R AF Ohmori, H Friedberg, EC Fuchs, RPP Goodman, MF Hanaoka, F Hinkle, D Kunkel, TA Lawrence, CW Livneh, Z Nohmi, T Prakash, L Prakash, S Todo, T Walker, GC Wang, ZG Woodgate, R TI The Y-family of DNA polymerases SO MOLECULAR CELL LA English DT Letter ID COMPILATION; ALIGNMENT C1 Kyoto Univ, Inst Virus Res, Sakyo Ku, Kyoto 6068507, Japan. Univ Texas, SW Med Ctr, Dept Pathol, Dallas, TX 75235 USA. CNRS, UPR 9003, ESBS, F-67400 Strasbourg, France. Univ So Calif, Dept Biol Sci & Chem, Los Angeles, CA 90089 USA. Osaka Univ, Inst Mol & Cellular Biol, Suita, Osaka 5650871, Japan. Japan Sci & Technol Corp, CREST, Suita, Osaka 5650871, Japan. RIKEN, Inst Phys & Chem Res, Wako, Saitama 3510198, Japan. Univ Rochester, Dept Biol, Rochester, NY 14627 USA. NIEHS, Res Triangle Pk, NC 27709 USA. Univ Rochester, Sch Med & Dent, Dept Biochem & Biophys, Rochester, NY 14642 USA. Weizmann Inst Sci, Dept Biol Chem, IL-76100 Rehovot, Israel. Natl Inst Hlth Sci, Div Genet & Mutagenesis, Setagaya Ku, Tokyo 1588501, Japan. Univ Texas, Med Branch, Sealy Ctr Mol Sci, Galveston, TX 77555 USA. Kyoto Univ, Radiat Biol Ctr, Kyoto 6068501, Japan. MIT, Dept Biol, Cambridge, MA 02139 USA. Univ Kentucky, Grad Ctr Toxicol, Lexington, KY 40536 USA. NICHHD, Sect DNA Replicat Repair & Mutagenesis, Bethesda, MD 20892 USA. RP Ohmori, H (reprint author), Kyoto Univ, Inst Virus Res, Sakyo Ku, Kyoto 6068507, Japan. RI Prakash, Satya/C-6420-2013; OI /0000-0001-7243-8261 NR 3 TC 590 Z9 604 U1 1 U2 15 PU CELL PRESS PI CAMBRIDGE PA 1100 MASSACHUSETTES AVE,, CAMBRIDGE, MA 02138 USA SN 1097-2765 J9 MOL CELL JI Mol. Cell PD JUL PY 2001 VL 8 IS 1 BP 7 EP 8 DI 10.1016/S1097-2765(01)00278-7 PG 2 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 456KP UT WOS:000170081900004 PM 11515498 ER PT J AU Allers, T Lichten, M AF Allers, T Lichten, M TI Intermediates of yeast meiotic recombination contain heteroduplex DNA SO MOLECULAR CELL LA English DT Article ID BREAK REPAIR MODEL; SACCHAROMYCES-CEREVISIAE; INITIATION SITE; GENE CONVERSION; MEIOSIS; IDENTIFICATION; MOLECULES; NDT80 AB The formation of heteroduplex DNA features prominently in all models for homologous recombination. A central intermediate in the current double-strand break repair model contains two Holliday junctions flanking a region of heteroduplex DNA. Studies of yeast meiosis have identified such intermediates but failed to detect associated heteroduplex DNA. We show here that these intermediates contain heteroduplex DNA, providing an important validation of the double-strand break repair model. However, we also detect intermediates where both Holliday junctions are to one side of the initiating DSB site, while the intervening region shows no evidence of heteroduplex DNA. Such structures are not easily accommodated by the canonical version of the double-strand break repair model. C1 NCI, Biochem Lab, Canc Res Ctr, Bethesda, MD 20892 USA. RP Lichten, M (reprint author), NCI, Biochem Lab, Canc Res Ctr, Bethesda, MD 20892 USA. RI Allers, Thorsten/G-5503-2010; Lichten, Michael/C-5795-2013 OI Allers, Thorsten/0000-0002-7308-3332; Lichten, Michael/0000-0001-9707-2956 NR 18 TC 87 Z9 88 U1 0 U2 6 PU CELL PRESS PI CAMBRIDGE PA 1100 MASSACHUSETTES AVE,, CAMBRIDGE, MA 02138 USA SN 1097-2765 J9 MOL CELL JI Mol. Cell PD JUL PY 2001 VL 8 IS 1 BP 225 EP 231 DI 10.1016/S1097-2765(01)00280-5 PG 7 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 456KP UT WOS:000170081900024 PM 11511375 ER PT J AU Yudt, MR Cidlowski, JA AF Yudt, MR Cidlowski, JA TI Molecular identification and characterization of A and B forms of the glucocorticoid receptor SO MOLECULAR ENDOCRINOLOGY LA English DT Article ID STEROID-HORMONE RECEPTORS; FACTOR-KAPPA-B; PROGESTERONE RECEPTORS; ANDROGEN RECEPTOR; MESSENGER-RNA; BETA-ISOFORM; DNA-BINDING; A-FORM; NUCLEAR; EXPRESSION AB The human glucocorticoid receptor (hGR alpha) is a ligand-activated transcription factor that mediates the physiological effects of corticosteroid hormones and is essential for life. Originally cloned in 1986, the transcriptionally active hGR alpha was reported to be a single protein species of 777 amino acids (molecular mass = 94 kDa). Biochemical data, obtained using various mammalian tissues and cell lines, however, have consistently revealed an additional, slightly smaller, second hGR protein (molecular mass = 91 kDa) that is not recognized by antibodies specific for the transcriptionally inactive and dominant negative, non-hormone-binding hGR beta isoform. We report here that when a single GR cDNA is transfected in COS-1 cells, or transcribed and translated in vitro, two forms of the receptor are observed, similar to those seen in cells that contain endogenous On. These data suggest that two forms of the hGR alpha are produced by alternative translation of the same gene and are henceforth termed GR-A and GR-B. To test this hypothesis, we have investigated the role of an internal ATG codon corresponding to methionine 27 (M27) as a potential alternative translation initiation site for the On. Mutagenesis of this ATG codon to ACG in human, rat, and mouse GR cDNA results in generation of a single 94-kDa protein species. On-A. Moreover, mutagenesis of the initial ATG codon to ACG (Met 1 to Thr) also resulted in production of single, shorter protein species (91 kDa), GR-B. Mutagenesis of the Kozak translation initiation sequence strongly indicates that a leaky ribosomal scanning mechanism is responsible for generating the GR-A and -B isoforms. Western blot analysis using peptide-specific antibodies show both the A and B receptor forms are present in human cell lines. Both receptors exhibit similar subcellular localization and nuclear translocation after ligand activation. Functional analyses of hGR-A and hGR-B under various glucocorticoid- responsive promoters reveal the shorter hGR-B to be nearly twice as effective as the longer hGR-A species in gene transactivation, but not in transrepression. C1 NIEHS, Lab Signal Transduct, NIH, Res Triangle Pk, NC 27709 USA. RP Cidlowski, JA (reprint author), NIEHS, Lab Integrat Biol, NIH, 111 TW Alexander Dr NC F307, Res Triangle Pk, NC 27709 USA. NR 46 TC 109 Z9 118 U1 0 U2 2 PU ENDOCRINE SOC PI BETHESDA PA 4350 EAST WEST HIGHWAY SUITE 500, BETHESDA, MD 20814-4110 USA SN 0888-8809 J9 MOL ENDOCRINOL JI Mol. Endocrinol. PD JUL PY 2001 VL 15 IS 7 BP 1093 EP 1103 DI 10.1210/me.15.7.1093 PG 11 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 448CJ UT WOS:000169609000004 PM 11435610 ER PT J AU Van Goor, F Zivadinovic, D Stojilkovic, SS AF Van Goor, F Zivadinovic, D Stojilkovic, SS TI Differential expression of ionic channels in rat anterior pituitary cells SO MOLECULAR ENDOCRINOLOGY LA English DT Article ID THYROTROPIN-RELEASING-HORMONE; ACTIVATED POTASSIUM CHANNELS; CALCIUM CHANNELS; CA2+ OSCILLATIONS; MEMBRANE CURRENTS; ACTION-POTENTIALS; K+ CHANNELS; SOMATOTROPHS; SECRETION; GONADOTROPES AB Secretory anterior pituitary cells are of the same origin, but exhibit cell type-specific patterns of spontaneous intracellular Ca2+ signaling and basal hormone secretion. To understand the underlying ionic mechanisms mediating these differences, we compared the ionic channels expressed in somatotrophs, lactotrophs, and gonadotrophs from randomly cycling female rats under identical cell culture and recording conditions, Our results indicate that a similar group of ionic channels are expressed in each cell type, including transient and sustained voltage-gated Ca2+ channels, tetrodotoxin-sensitive Na+ channels, transient and delayed rectifying K+ channels, and multiple Ca2+ sensitive K+ channel subtypes. However, there were marked differences in the expression levels of some of the ionic channels. Specifically, lactotrophs and somatotrophs exhibited low expression levels of tetrodotoxin-sensitive Na+ channels end high expression levels of the large-conductance, Ca2+-activated K+ channel compared with those observed in gonadotrophs. In addition, functional expression of the transient K+ channel was much higher in lactotrophs and gonadotrophs than in somatotrophs. Finally, the expression of the transient voltage-gated Ca2+ channels was higher in somatotrophs than in lactotrophs and gonadotrophs. These results indicate that there are cell type-specific patterns of ionic channel expression, which may be of physiological significance for the control of Ca2+ homeostasis and secretion in unstimulated and receptor-stimulated anterior pituitary cells. C1 NICHHD, Endocrinol & Reprod Res Branch, Sect Cellular Signaling, NIH, Bethesda, MD 20892 USA. RP Van Goor, F (reprint author), NICHHD, Endocrinol & Reprod Res Branch, Sect Cellular Signaling, NIH, Bldg 49,Room 6A36,49 Convent Dr, Bethesda, MD 20892 USA. NR 54 TC 48 Z9 48 U1 0 U2 3 PU ENDOCRINE SOC PI BETHESDA PA 4350 EAST WEST HIGHWAY SUITE 500, BETHESDA, MD 20814-4110 USA SN 0888-8809 J9 MOL ENDOCRINOL JI Mol. Endocrinol. PD JUL PY 2001 VL 15 IS 7 BP 1222 EP 1236 DI 10.1210/me.15.7.1222 PG 15 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 448CJ UT WOS:000169609000015 PM 11435620 ER PT J AU Donovan, PJ Smith, GT Riggs, CW AF Donovan, PJ Smith, GT Riggs, CW TI Hamster and rat fetal cells have low spontaneous mutation frequencies and rates SO MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS LA English DT Article DE in vivo; 6-thioguanine; diphtheria toxin; ouabain; 2,6-diaminopurine ID GENE-TOX PROGRAM; SYRIAN-HAMSTER; OVARY CELLS; DNA GLYCOSYLASE; CYTO-TOXICITY; EMBRYO CELLS; REPAIR; MUTAGENESIS; EXPRESSION; CULTURE AB Somatic cells of whole Syrian hamster fetuses (gestation day 13) were isolated and tested by an in vivo/in vitro mutation assay for spontaneous mutation frequencies using independent 6-thioguanine (6-TG), diphtheria toxin (DT), and ouabain mutation selection systems. Optimum conditions were ascertained. For 6-TG mutants, a total of 21 mutants were found in cells from 24 litters on 1993 plates, for an overall mutant frequency of 1.8 x 10(-7) per viable cell with 12 positive litters. In all, 26 litters were tested using DT; 77 mutants were found in 840 plates, yielding an overall mutant frequency of 2.6 x 10(-7), with 20 positive Litters. No correlations or familial effects were found among 23 Litters tested for both DT and 6-TG. Of 14 litters which were tested for ouabain mutants, 4 were positive, with a total of 5 mutants found on 988 plates, for an overall mutant frequency of 7.6 x 10(-8). For 14 F344 rat fetuses, the overall 6-TG spontaneous mutation frequency was determined to be 1.6 x 10(-7). From the data, estimates of mutation rates were calculated. For mutation to 6-TG resistance the rate was 8.3 x 10(-8) for mutation to DT resistance the rate was 8.1 x 10(-8) and for ouabain, the spontaneous mutation rate was 5.7 x 10(-8). For F344 rat, the spontaneous mutation rate was 1.1 x 10(-7). Induced mutant frequencies after in utero exposure to 1 mmol/kg N-ethyl-N-nitrosourea (ENU) were 311, 135 and 200 times the spontaneous value for 6-TG, DT and ouabain, respectively, for Syrian hamster fetal cells and 125 times the spontaneous 6-TG value for fetal F344 rat cells. Both spontaneous mutation frequencies and underlying spontaneous mutation rates are low, consistent with the view that fetal cells exercise extremely tight control over DNA fidelity. Published by Elsevier Science B.V. C1 NCI, Comparat Carcinogenesis Lab, Ft Detrick, MD 21702 USA. NCI, Data Management Serv Inc, Frederick, MD 21702 USA. RP Donovan, PJ (reprint author), NCI, Comparat Carcinogenesis Lab, Bldg 538,Room 205E, Ft Detrick, MD 21702 USA. FU NCI NIH HHS [N01-CO-46002] NR 52 TC 5 Z9 5 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0027-5107 J9 MUTAT RES-FUND MOL M JI Mutat. Res.-Fundam. Mol. Mech. Mutagen. PD JUL 1 PY 2001 VL 478 IS 1-2 BP 51 EP 63 DI 10.1016/S0027-5107(01)00107-5 PG 13 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology SC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology GA 449EV UT WOS:000169672100004 PM 11406169 ER PT J AU Pavlov, YI Nguyen, D Kunkel, TA AF Pavlov, YI Nguyen, D Kunkel, TA TI Mutator effects of overproducing DNA polymerase eta (Rad30) and its catalytically inactive variant in yeast SO MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS LA English DT Article DE polymerase eta; spontaneous mutagenesis; mismatch repair; exonucleolytic proofreading; replication fidelity ID BASE ANALOG 6-N-HYDROXYLAMINOPURINE; ESCHERICHIA-COLI DINB; SACCHAROMYCES-CEREVISIAE; XERODERMA-PIGMENTOSUM; MISMATCH REPAIR; THYMINE DIMER; GENE ENCODES; MUTAGENESIS; REPLICATION; MUTATIONS AB DNA polymerase eta synthesizes DNA in vitro with low fidelity. Based on this, here we report the effects of deletion or increased expression of yeast RAD30 gene, encoding for polymerase eta (Pol eta), on spontaneous mutagenesis in vivo. Deletion of RAD30 did not affect spontaneous mutagenesis. Overproduction of Rad30p was slightly mutagenic in a wild-type yeast strain and moderately mutagenic in strains with inactive 3 ' --> 5 ' -exonuclease of DNA polymerase epsilon or DNA mismatch repair. These data suggest that excess Rad30p reduces replication fidelity in vivo and that the induced errors may be corrected by exonucleolytic proofreading and DNA mismatch repair. However, the magnitude of mutator effect (only up to 10-fold) suggests that the replication fork is protected from inaccurate synthesis by Pol eta in the absence of DNA damage. Overproduction of catalytically inactive Rad30p was also mutagenic, suggesting that much of the mutator effect results from indirect perturbation of replication rather than from direct misincorporation by Po eta. Moreover, while excess wild-type Pol eta primarily induced base substitutions in the msh6 and pms1 strains, excess inactive Rad30p induced both base substitutions and frameshifts. This suggests that more than one mutagenic mechanism is operating when RAD30 is overexpressed. Published by Elsevier Science B.V. C1 NIEHS, Mol Genet Lab, Res Triangle Pk, NC 27709 USA. NIEHS, Struct Biol Lab, Res Triangle Pk, NC 27709 USA. RP Pavlov, YI (reprint author), NIEHS, Mol Genet Lab, Res Triangle Pk, NC 27709 USA. NR 44 TC 36 Z9 36 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0027-5107 J9 MUTAT RES-FUND MOL M JI Mutat. Res.-Fundam. Mol. Mech. Mutagen. PD JUL 1 PY 2001 VL 478 IS 1-2 BP 129 EP 139 DI 10.1016/S0027-5107(01)00131-2 PG 11 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology SC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology GA 449EV UT WOS:000169672100012 PM 11406177 ER PT J AU Xu, XL Qiao, WH Linke, SP Cao, L Li, WM Furth, PA Harris, CC Deng, CX AF Xu, XL Qiao, WH Linke, SP Cao, L Li, WM Furth, PA Harris, CC Deng, CX TI Genetic interactions between tumor suppressors Brca1 and p53 in apoptosis, cell cycle and tumorigenesis SO NATURE GENETICS LA English DT Letter ID DAMAGE-INDUCED PHOSPHORYLATION; ATM-DEPENDENT PHOSPHORYLATION; DOUBLE-STRAND BREAKS; DNA-DAMAGE; GAMMA-IRRADIATION; MOUSE; MICE; PROTEIN; MDM2; MORPHOGENESIS AB Breast cancer is a chief cause of cancer-related mortality that affects women worldwide. About 8% of cases are hereditary. and approximately half of these are associated with germline mutations of the breast tumor suppressor gene BRCA1 (refs. 1,2). We have previously reported a mouse model in which Brca1 exon 11 is eliminated in mammary epithelial cells through Cre-mediated excision(3). This mutation is often accompanied by alterations in transformation-related protein 53 (Trp53, encoding p53), which substantially accelerates mammary tumor formation. Here. we sought to elucidate the underlying mechanism(s) using mice deficient in the Brca1 exon 11 isoform (Brca1(Delta 11/Delta 11)). Brca1(Delta 11/Delta 11) embryos died late in gestation because of widespread apoptosis. Unexpectedly. elimination of one Trp53 allele completely rescues this embryonic lethality and restores normal mammary gland development. However, most female Brca1(Delta 11/Delta 11)Trp53(+/-) mice develop mammary tumors with loss of the remaining Trp53 allele within 6-12 months. Lymphoma and ovarian tumors also occurr at lower frequencies. Heterozygous mutation of Trp53 decreases p53 and results in attenuated apoptosis and G(1)-S checkpoint control, allowing Brca1(Delta 11/Delta 11) cells to proliferate. The p53 protein regulates Brca1 transcription both in vitro and in vivo, and Brca1 participates in p53 accumulation after gamma -irradiation through regulation of its phosphorylation and Mdm2 expression. These findings provide a mechanism for BRCA1-associated breast carcinogenesis. C1 NIDDKD, Genet Dev & Dis Branch, NIH, Bethesda, MD 20892 USA. NCI, Human Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. Univ Maryland, Sch Med, Dept Med, Inst Human Virol, Baltimore, MD 21201 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 30 TC 231 Z9 238 U1 2 U2 8 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1061-4036 J9 NAT GENET JI Nature Genet. PD JUL PY 2001 VL 28 IS 3 BP 266 EP 271 DI 10.1038/90108 PG 6 WC Genetics & Heredity SC Genetics & Heredity GA 448XZ UT WOS:000169656400020 PM 11431698 ER PT J AU Yang, JH Lin, Y Guo, ZJ Cheng, JK Huang, JY Deng, L Liao, W Chen, ZJ Liu, ZG Su, B AF Yang, JH Lin, Y Guo, ZJ Cheng, JK Huang, JY Deng, L Liao, W Chen, ZJ Liu, ZG Su, B TI The essential role of MEKK3 in TNF-induced NF-kappa B activation SO NATURE IMMUNOLOGY LA English DT Article ID INDUCED CELL-DEATH; KINASE-ALPHA; INDUCED APOPTOSIS; RIP; BETA; IKK; PROTEIN; COMPLEX; PHOSPHORYLATION; RECRUITMENT AB Activation of I kappaB kinase (IKK) is the key step in stimulation of the transcription factor NF-kappaB, which regulates many genes in the inflammatory response pathway. The molecular mechanism that underlies IKK activation in response to tumor necrosis factor (TNF) is still unknown. Using mitogen-activated protein kinase kinase kinase 3 (MEKK3)-deficient fibroblast cells, we found that MEKK3 plays a critical role in TNF-induced NF-kappaB activation. We have shown that MEKK3 is required for IKK activation and functions downstream of receptor-interacting protein (RIP) and TNF receptor-associated factor 2. We have also shown that MEKK3 interacts with RIP and directly phosphorylates IKK. The kinase activity of MEKK3 is pivotal to its function and, therefore, MEKK3 links RIP and IKK in TNF-induced NF-kappaB activation. C1 Univ Texas, MD Anderson Canc Ctr, Dept Immunol, Houston, TX 77030 USA. Univ Texas, MD Anderson Canc Ctr, Dept Biochem & Mol Biol, Houston, TX 77030 USA. NCI, Dept Cell & Canc Biol, Med Branch, Div Clin Sci,NIH, Bethesda, MD 20892 USA. Univ Texas, SW Med Ctr, Dept Mol Biol, Dallas, TX 75235 USA. RP Su, B (reprint author), Univ Texas, MD Anderson Canc Ctr, Dept Immunol, Houston, TX 77030 USA. FU NCI NIH HHS [CA16672]; NIAID NIH HHS [AI44016] NR 31 TC 271 Z9 289 U1 2 U2 15 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1529-2908 J9 NAT IMMUNOL JI Nat. Immunol. PD JUL PY 2001 VL 2 IS 7 BP 620 EP 624 DI 10.1038/89769 PG 5 WC Immunology SC Immunology GA 449JU UT WOS:000169682100015 PM 11429546 ER PT J AU Kirn, D Martuza, RL Zwiebel, J AF Kirn, D Martuza, RL Zwiebel, J TI Replication-selective virotherapy for cancer: Biological principles, risk management and future directions SO NATURE MEDICINE LA English DT Review ID NEWCASTLE-DISEASE VIRUS; HERPES-SIMPLEX VIRUS; PHASE-I TRIAL; PROSTATE-SPECIFIC ANTIGEN; TUMOR-CELL VACCINE; GENE-THERAPY; COMPETENT ADENOVIRUS; NECK-CANCER; ATTENUATED ADENOVIRUS; ONCOLYTIC ADENOVIRUS AB In the search for novel cancer therapies that can be used in conjunction with existing treatments, one promising area of research is the use of viral vectors and whole viruses. This review describes the underlying biological principles and current status of the field, outlines approaches for improving clinical effectiveness and discusses the unique safety and regulatory issues surrounding viral therapies. C1 Univ London Imperial Coll Sci Technol & Med, Viral & Genet Therapy Programme, Sch Med, London, England. Hammersmith Hosp, Imperial Canc Res Fund, London, England. Harvard Univ, Sch Med, Boston, MA USA. Massachusetts Gen Hosp, Boston, MA 02114 USA. NCI, Div Canc Treatment, Canc Therapy Evaluat Program, Bethesda, MD 20892 USA. RP Kirn, D (reprint author), Univ London Imperial Coll Sci Technol & Med, Viral & Genet Therapy Programme, Sch Med, London, England. FU NINDS NIH HHS [NS32677] NR 78 TC 286 Z9 304 U1 8 U2 42 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1078-8956 J9 NAT MED JI Nat. Med. PD JUL PY 2001 VL 7 IS 7 BP 781 EP 787 DI 10.1038/89901 PG 7 WC Biochemistry & Molecular Biology; Cell Biology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Cell Biology; Research & Experimental Medicine GA 451NR UT WOS:000169808600028 PM 11433341 ER PT J AU Batkai, S Jarat, Z Wagner, JA Goparaju, SK Varga, K Liu, J Wang, L Mirshahi, F Khanolkar, AD Makriyannis, A Urbaschek, R Garcia, N Sanyal, AJ Kunos, G AF Batkai, S Jarat, Z Wagner, JA Goparaju, SK Varga, K Liu, J Wang, L Mirshahi, F Khanolkar, AD Makriyannis, A Urbaschek, R Garcia, N Sanyal, AJ Kunos, G TI Endocannabinoids acting at vascular CB, receptors mediate the vasodilated state in advanced liver cirrhosis SO NATURE MEDICINE LA English DT Article ID NITRIC-OXIDE SYNTHASE; HYPERDYNAMIC CIRCULATION; CANNABINOID RECEPTORS; PORTAL-HYPERTENSION; ENDOTHELIAL-CELLS; ENDOTOXIN LEVELS; WATER-RETENTION; RATS; ANANDAMIDE; RESISTANCE AB Advanced cirrhosis is associated with generalized vasodilation of unknown origin, which contributes to mortality. Cirrhotic patients are endotoxemic, and activation of Vascular cannabinoid CB1 receptors has been implicated in endotoxin-induced hypotension. Here we show that rats with biliary cirrhosis have low blood pressure, which is elevated by the CB1 receptor antagonist SR141716A. The low blood pressure of rats with CCl4-induced cirrhosis was similarly reversed by SR141716A, which also reduced the elevated mesenteric blood flow and portal pressure. Monocytes from cirrhotic but not control patients or rats elicited SR141716A-sensitive hypotension in normal recipient rats and showed significantly elevated levels of anandamide. Compared with non-cirrhotic controls, in cirrhotic human livers there was a three-fold increase in CB1 receptors on isolated vascular endothelial cells. These results implicate anandamide and vascular CB1 receptors in the vasodilated state in advanced cirrhosis and indicate a novel approach for its management. C1 NIAAA, NIH, Bethesda, MD USA. Virginia Commonwealth Univ, Dept Med, Richmond, VA 23298 USA. Univ Connecticut, Sch Pharm, Dept Pharmaceut Sci, Storrs, CT USA. Univ Heidelberg, Klinikum Mannheim, Inst Med Microbiol & Hyg, Mannheim, Germany. RP Kunos, G (reprint author), NIAAA, NIH, Bethesda, MD USA. RI Batkai, Sandor/G-3889-2010; Goparaju, Sravan/H-3422-2011; Batkai, Sandor/H-7983-2014 FU NCRR NIH HHS [MO1-RR-00065-38]; NHLBI NIH HHS [HL-59257] NR 43 TC 241 Z9 247 U1 0 U2 2 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1078-8956 J9 NAT MED JI Nat. Med. PD JUL PY 2001 VL 7 IS 7 BP 827 EP 832 DI 10.1038/89953 PG 6 WC Biochemistry & Molecular Biology; Cell Biology; Medicine, Research & Experimental SC Biochemistry & Molecular Biology; Cell Biology; Research & Experimental Medicine GA 451NR UT WOS:000169808600035 PM 11433348 ER PT J AU Bennink, ML Leuba, SH Leno, GH Zlatanova, J de Grooth, BG Greve, J AF Bennink, ML Leuba, SH Leno, GH Zlatanova, J de Grooth, BG Greve, J TI Unfolding individual nucleosomes by stretching single chromatin fibers with optical tweezers SO NATURE STRUCTURAL BIOLOGY LA English DT Article ID DNA; FORCE; MOLECULE; HISTONE; TRANSCRIPTION; POLYMERASE; TEMPLATE; PROTEIN; TENSION; B4 AB Single chromatin fibers were assembled directly in the flow cell of an optical tweezers setup. A single h phage DNA molecule, suspended between two polystyrene beads, was exposed to a Xenopus laevis egg extract, leading to chromatin assembly with concomitant apparent shortening of the DNA molecule. Assembly was force-dependent and could not take place at forces exceeding 10 pN, The assembled single chromatin fiber was subjected to stretching by controlled movement of one of the beads with the force generated in the molecule continuously monitored with the second bead trapped in the optical trap. The force displayed discrete, sudden drops upon fiber stretching, reflecting discrete opening events in fiber structure. These opening events were quantized at increments in fiber length of similar to 65 mn and are attributed to unwrapping of the DNA from around individual histone octamers, Repeated stretching and relaxing of the fiber in the absence of egg extract showed that the loss of histone octamers was irreversible. The forces measured for Individual nucleosome disruptions are in the range of 20-40 pN, comparable to forces reported for RNA- and DNA-polymerases. C1 Univ Twente, Dept Appl Phys, iBME Res Inst, NL-7500 AE Enschede, Netherlands. NCI, Lab Recptor Biol & Gene Express, NIH, Bethesda, MD 20892 USA. Univ Mississippi, Med Ctr, Dept Biochem, Jackson, MS 39216 USA. Polytech Univ, Dept Chem Engn Chem & Mat Sci, MetroTech Ctr 6, Brooklyn, NY 11201 USA. RP Bennink, ML (reprint author), Univ Twente, Dept Appl Phys, iBME Res Inst, POB 217, NL-7500 AE Enschede, Netherlands. RI Zlatanova, Jordanka/B-3273-2009 NR 31 TC 180 Z9 181 U1 5 U2 26 PU NATURE AMERICA INC PI NEW YORK PA 345 PARK AVE SOUTH, NEW YORK, NY 10010-1707 USA SN 1072-8368 J9 NAT STRUCT BIOL JI Nat. Struct. Biol. PD JUL PY 2001 VL 8 IS 7 BP 606 EP 610 DI 10.1038/89646 PG 5 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 446JE UT WOS:000169509500015 PM 11427891 ER PT J AU Kosel, S Grasbon-Frodl, EM Arima, K Chimelli, L Hahn, M Hashizume, Y Hulette, C Ikeda, K Jacobsen, PF Jones, M Kobayashi, M Love, S Mizutani, T Rosemberg, S Sasaki, A Smith, TW Takahashi, H Vortmeyer, AO Graeber, MB AF Kosel, S Grasbon-Frodl, EM Arima, K Chimelli, L Hahn, M Hashizume, Y Hulette, C Ikeda, K Jacobsen, PF Jones, M Kobayashi, M Love, S Mizutani, T Rosemberg, S Sasaki, A Smith, TW Takahashi, H Vortmeyer, AO Graeber, MB TI Inter-laboratory comparison of DNA preservation in archival paraffin-embedded human brain tissue from participating centres on four continents SO NEUROGENETICS LA English DT Article DE DNA preservation; HPLC analysis; mitochondrial DNA; neurodegeneration; PCR inhibitor; ultrafiltration ID POLYMERASE-CHAIN-REACTION; PARKINSON-DISEASE; PCR AMPLIFICATION; VIRUS-DNA; EXTRACTION; MUTATIONS; HAPLOGROUPS; INHIBITOR; APOPTOSIS; FIXATION AB DNA extracted from formalin-fixed and paraffin-embedded brain tissue is known to contain as yet ill-characterized inhibitors of the PCR process. As part of a project that aims to clarify the role of mitochondrial DNA sequence variation in human neurodegenerative diseases using DNA from various ethnic backgrounds, we have investigated factors that influence the preservation of archival DNA and its suitability for PCR. In this study, neuropathological tissue samples were analysed that had been routinely processed in 18 international centres on four continents. Following DNA extraction, PCR amplification of mitochondrial and nuclear DNA sequences was performed with and without additional purification of the template DNA. In addition, the DNA used for PCR was analysed by HPLC. Phosphate-buffered formalin proved to be a superior fixative compared with unbuffered aldehyde: DNA extraction resulted in greater yields, the molecular weight of the isolated DNA was higher and PCR was more successful. PCR inhibitors were identified as (1) high concentrations of small (<300 bp) DNA fragments that competitively compete with template DNA and (2) contaminants of the DNA template solution including denatured protein that cannot be completely removed by phenolic extraction. HPLC analysis did not reveal significant qualitative differences between DNA isolated from fresh-frozen tissue samples and DNA recovered from formalin-fixed, paraffin-embedded brain tissue. The fact that DNA could be amplified from the majority of tissue specimens in this study suggests that rare diseases and diseases where ethnic background plays an important role can be sampled for genetic polymorphism analysis on a global scale using archival neuropathological collections. C1 Imperial Coll Sch Med, Dept Neuropathol, London W6 8RF, England. Univ Munich, Inst Neuropathol, Mol Neuropathol Lab, Munich, Germany. NCNP, Natl Ctr Hosp Mental Nervous & Muscular Dis, Dept Lab Med, Tokyo, Japan. Univ Fed Rio de Janeiro, Dept Pathol, BR-21941 Rio De Janeiro, Brazil. Univ Fed Fluminense, Sch Med, Dept Pathol, Niteroi, RJ, Brazil. Aichi Med Univ, Inst Med Sci Aging, Aichi, Japan. Duke Univ, Med Ctr, Bryan Alzheimers Dis Res Ctr, Durham, NC USA. Tokyo Inst Psychiat, Dept Neuropathol, Tokyo, Japan. Royal Perth Hosp, Dept Neuropathol, Perth, WA, Australia. Hosp Ninos Sor Maria Ludovica, Dept Pathol, La Plata, Buenos Aires, Argentina. Tokyo Womens Med Coll, Dept Pathol, Tokyo 162, Japan. Frenchay Hosp, Dept Neuropathol, Bristol, Avon, England. Nihon Univ, Sch Med, Dept Neurol, Tokyo, Japan. Univ Sao Paulo, Sch Med, Dept Pathol, Div Neuropathol, Sao Paulo, Brazil. Gunma Univ, Sch Med, Dept Pathol, Gunma, Japan. Univ Massachusetts, Dept Pathol, Worcester, MA 01605 USA. Niigata Univ, Inst Brain Res, Dept Pathol, Niigata, Japan. NINDS, Mol Pathogenesis Lab, Bethesda, MD 20892 USA. RP Graeber, MB (reprint author), Imperial Coll Sch Med, Dept Neuropathol, Charing Cross Campus,Fulham Palace Rd, London W6 8RF, England. RI Graeber, Manuel/H-3224-2011; Love, Seth/E-6545-2012; Rosemberg, Sergio/F-4464-2012 NR 28 TC 13 Z9 15 U1 0 U2 4 PU SPRINGER-VERLAG PI NEW YORK PA 175 FIFTH AVE, NEW YORK, NY 10010 USA SN 1364-6745 J9 NEUROGENETICS JI Neurogenetics PD JUL PY 2001 VL 3 IS 3 BP 163 EP 170 PG 8 WC Genetics & Heredity; Clinical Neurology SC Genetics & Heredity; Neurosciences & Neurology GA 457KM UT WOS:000170136200008 PM 11523568 ER PT J AU Weeks, RA Honda, M Catalan, MJ Hallett, M AF Weeks, RA Honda, M Catalan, MJ Hallett, M TI Comparison of auditory, somatosensory, and visually instructed and internally generated finger movements: A PET study SO NEUROIMAGE LA English DT Article ID POSITRON EMISSION TOMOGRAPHY; POSTERIOR CINGULATE CORTEX; EXTERNALLY TRIGGERED MOVEMENTS; SUPPLEMENTARY MOTOR AREA; CEREBRAL BLOOD-FLOW; PREMOTOR CORTEX; FRONTAL-CORTEX; BASAL GANGLIA; NEURONAL-ACTIVITY; CORTICAL AREAS AB We sought to determine how the pattern of cerebral activation, and in particular in frontal motor areas, during the performance of conditional motor tasks is dependent upon the modality of instruction (visual, auditory, or somatosensory). Regional cerebral blood flow (rCBF) changes with externally instructed movements were also compared with internally generated, self-paced, movements. We used positron emission tomography (PET) with the tracer (H2O)-O-15 to measure rCBF in 22 healthy volunteers, External stimuli consisted of the randomized presentation of single or double impulses using a single modality for each condition. In the movement scans, the subjects used the index and middle fingers of their right hands to press a left button for a single and a right button for a double impulse, respectively. In the control scans, subjects were required to covertly distinguish a single from a double stimulus without a motor response. Data were analyzed using conventional subtraction techniques with a statistical threshold of Z > 2.33 with corrections for multiple comparisons. When the activation differences between the three externally instructed movement conditions were statistically compared, nonsignificant trends toward increased rCBF in the sensory cortex of the modality of the cue were observed but no differential activity in cortical motor areas. Internally generated movements, when compared to externally triggered movements, were associated with enhanced activation in bilateral medial and lateral premotor, dorsolateral prefrontal and superior parietal regions, largely confirming previous reports. The data indicate that, on a regional level, modality-specific processing in a conditional motor task does not occur in frontal motor areas and is probably confined to sensory areas. (C) 2001 Academic Press. C1 NINDS, Human Motor Control Sect, Med Neurol Branch, NIH, Bethesda, MD 20892 USA. RP Weeks, RA (reprint author), NINDS, Human Motor Control Sect, Med Neurol Branch, NIH, Bethesda, MD 20892 USA. NR 43 TC 44 Z9 44 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 1053-8119 J9 NEUROIMAGE JI Neuroimage PD JUL PY 2001 VL 14 IS 1 BP 219 EP 230 DI 10.1006/nimg.2001.0780 PN 1 PG 12 WC Neurosciences; Neuroimaging; Radiology, Nuclear Medicine & Medical Imaging SC Neurosciences & Neurology; Radiology, Nuclear Medicine & Medical Imaging GA 446DQ UT WOS:000169498000022 PM 11525332 ER PT J AU Ruskin, DN Bergstrom, DA Baek, D Freeman, LE Walters, JR AF Ruskin, DN Bergstrom, DA Baek, D Freeman, LE Walters, JR TI Cocaine or selective block of dopamine transporters influences multisecond oscillations in firing rate in the globus pallidus SO NEUROPSYCHOPHARMACOLOGY LA English DT Article DE cocaine; dopamine; GBR-12909; globus pallidus; NMDA receptor; oscillation; wavelet ID SUBTHALAMIC NUCLEUS; CA2+ OSCILLATIONS; 6-HYDROXYDOPAMINE-LESIONED RAT; RECEPTOR AGONISTS; UPTAKE INHIBITION; SQUIRREL-MONKEYS; SUBSTANTIA-NIGRA; GENE-EXPRESSION; BASAL GANGLIA; IN-VIVO AB Previous studies have shown that direct-acting dopamine agonists modulate the multisecond oscillations which are present in globus pallidus spike trains in vivo in awake rats. To investigate possible modulation by endogenous dopamine and by other monoamines, and by drugs with abuse potential, cocaine or selective monoamine uptake blockers were injected systemically during extracellular recording of single globus pallidus neurons and the results analyzed with spectral and wavelet methods. Both cocaine and the selective dopamine uptake blocker GBR-12909 significantly shortened the period of multisecond oscillations, as well as increasing overall firing rate. Cocaine effects were blocked by dopamine antagonist pretreatment, as well as by N-methyl-D-aspartate receptor antagonist (MK-801) pretreatment. Desipramine and fluoxetine (blockers of norepinephrine and serotonin uptake, respectively) had no significant effects on multisecond oscillations. The results suggest that dopamine has a primary role among monoamines in modulating multisecond oscillations in globus pallidus activity, and that tonic dopaminergic and glutamatergic transmission is necessary for normal slow oscillatory function. [Neuropsychopharmacology 25:28-40, 2001] (C) 2001 American College of Neuropsychopharmacology. Published by Elsevier Science Inc. C1 NINDS, Expt Therapeut Branch, NIH, Bethesda, MD 20892 USA. RP Ruskin, DN (reprint author), 10 Ctr Dr,Room 5C103, Bethesda, MD 20892 USA. NR 56 TC 18 Z9 19 U1 0 U2 0 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0893-133X J9 NEUROPSYCHOPHARMACOL JI Neuropsychopharmacology PD JUL PY 2001 VL 25 IS 1 BP 28 EP 40 DI 10.1016/S0893-133X(00)00241-4 PG 13 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA 437YH UT WOS:000169026600002 PM 11377917 ER PT J AU Sora, I Elmer, G Funada, M Pieper, J Li, XF Hall, FS Uhl, GR AF Sora, I Elmer, G Funada, M Pieper, J Li, XF Hall, FS Uhl, GR TI mu opiate receptor gene dose effects on different morphine actions: Evidence for differential in vivo mu receptor reserve SO NEUROPSYCHOPHARMACOLOGY LA English DT Article DE transgenic knockout mice; locomotion; conditioned place preference; tolerance; physical dependence; lethality; self-administration ID DELTA-OPIOID RECEPTOR; G-PROTEIN ACTIVATION; KNOCKOUT MICE; INDUCED ANALGESIA; DEFICIENT MICE; MEDIATED ANTINOCICEPTION; PLACE PREFERENCE; AGONISTS; MECHANISMS; EXPRESSION AB Homozygous transgenic knockout mice without mu -opioid receptors lack morphine-induced antinociception, locomotion, tolerance, physical dependence, and reward. mu receptors thus appear to play central roles in these morphine actions. Different levels of mu receptor expression are found in different humans and in different animal strains. In vitro studies indicate that some morphine responses persist after inactivation of as many as 90% of the initial mu receptor complement, while others are attenuated after inactivating many fewer receptors. Varying levels of mu receptor reserve could thus exist in different mu -expressing neuronal populations in vivo. Heterozygous mu receptor knockout mice express half of wild-type mu receptor levels. Tests of morphine actions in these mice reveal evidence for differing mu receptor reserves in brain circuits that mediate distinct opiate effects. Heterozygotes display attenuated locomotion, reduced morphine self-administration, intact tolerance, rightward shifts in morphine lethality dose/effect relationships, and variable effects on place preference compared to wild-type mice. They demonstrate full physical dependence, as measured by naloxone-precipitated abstinence following five days of morphine administration. Neuroadaptive changes in sites other than mu receptors could be involved in some of these results. Nevertheless, these data document substantial influences that individual differences in levels of mu receptor expression could exert on distinct opiate drug effects. They support the idea that functional mu receptor reserve differs among the diverse neuronal populations that mediate distinct properties of opiate drugs. [Neuropsychopharmacology 25:41-54, 2001] (C) 2001 American College of Neuropsychopharmacology. Published by Elsevier Science Inc. C1 NIDA, Mol Neurobiol Branch, IRP, NIH, Baltimore, MD 21224 USA. UMAB, Maryland Psychiat Res Ctr, Catonsville, MD 21228 USA. Natl Ctr Neurol & Psychiat, Natl Inst Mental Hlth, Div Drug Dependence Res, Chiba, Japan. RP NIDA, Mol Neurobiol Branch, IRP, NIH, POB 5180,5500 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Hall, Frank/C-3036-2013 OI Hall, Frank/0000-0002-0822-4063 FU NIDA NIH HHS [DA11185] NR 51 TC 91 Z9 95 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI LONDON PA MACMILLAN BUILDING, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0893-133X EI 1740-634X J9 NEUROPSYCHOPHARMACOL JI Neuropsychopharmacology PD JUL PY 2001 VL 25 IS 1 BP 41 EP 54 DI 10.1016/S0893-133X(00)00252-9 PG 14 WC Neurosciences; Pharmacology & Pharmacy; Psychiatry SC Neurosciences & Neurology; Pharmacology & Pharmacy; Psychiatry GA 437YH UT WOS:000169026600003 PM 11377918 ER PT J AU Gahl, WA AF Gahl, WA TI New therapies for Fabry's disease. SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Editorial Material ID GALACTOSIDASE C1 NIH, Bethesda, MD 20892 USA. RP Gahl, WA (reprint author), NIH, Bldg 10, Bethesda, MD 20892 USA. NR 12 TC 11 Z9 11 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 JUL 1 PY 2001 VL 345 IS 1 BP 55 EP 57 DI 10.1056/NEJM200107053450109 PG 4 WC Medicine, General & Internal SC General & Internal Medicine GA 449QJ UT WOS:000169697600009 PM 11439950 ER PT J AU Ortega, R Deves, G Bonnin-Mosbah, M Salome, M Susini, J Anderson, LM Kasprzak, KS AF Ortega, R Deves, G Bonnin-Mosbah, M Salome, M Susini, J Anderson, LM Kasprzak, KS TI Chromium mapping in male mice reproductive glands exposed to CrCl3 using proton and X-ray synchrotron radiation microbeams SO NUCLEAR INSTRUMENTS & METHODS IN PHYSICS RESEARCH SECTION B-BEAM INTERACTIONS WITH MATERIALS AND ATOMS LA English DT Article; Proceedings Paper CT 7th International Conference on Nuclear Microprobe Technology and Applications CY SEP 10-15, 2000 CL BORDEAUX, FRANCE SP CNRS, CEA, CNRS Univ Bordeaux, CNRS Minist Culture & Commun DE nuclear microprobe; synchrotron radiation; X-ray fluorescence; chromium; carcinogenesis ID OVARIAN ADENOCARCINOMA CELLS; CISPLATIN-RESISTANT; MICROANALYSIS; PLATINUM; TISSUE AB Preconception exposure to certain chemicals may increase risk of tumors in offspring, especially with regard to occupational metals such as chromium. However, the mechanism of chromium trans-generation carcinogenicity remains unknown. Using scanning proton X-ray microanalysis we have been able to detect chromium in testicular tissue sections from mice treated by intraperitoneal injection of I mmol/kg CrCl3. Chromium concentration was about 5 mug/g dry mass in average, but higher concentrations were found within the limiting membrane of the testes, the tunica albuginea. In addition, synchrotron radiation X-ray fluorescence measurements, with microscopic resolution, clearly demonstrated the presence of chromium in the tunica albuginea but also within isolated cells from the interstitial connective tissue. (C) 2001 Elsevier Science B.V. All rights reserved. C1 Univ Bordeaux 1, CNRS UMR 5084, LCNAB, F-33175 Gradignan, France. CE Saclay, CEA, CNRS, LPS, F-91191 Gif Sur Yvette, France. ESRF, F-38043 Grenoble, France. NCI, Comparat Carcinogenesis Lab, Frederick, MD 21702 USA. RP Ortega, R (reprint author), Univ Bordeaux 1, CNRS UMR 5084, LCNAB, BP 120 Haut Vigneau, F-33175 Gradignan, France. NR 9 TC 13 Z9 13 U1 0 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0168-583X J9 NUCL INSTRUM METH B JI Nucl. Instrum. Methods Phys. Res. Sect. B-Beam Interact. Mater. Atoms PD JUL PY 2001 VL 181 BP 485 EP 488 DI 10.1016/S0168-583X(01)00477-3 PG 4 WC Instruments & Instrumentation; Nuclear Science & Technology; Physics, Atomic, Molecular & Chemical; Physics, Nuclear SC Instruments & Instrumentation; Nuclear Science & Technology; Physics GA 470RR UT WOS:000170885400085 ER PT J AU Eckelman, WC AF Eckelman, WC TI Radiolabeled muscarinic radioligands for in vivo studies SO NUCLEAR MEDICINE AND BIOLOGY LA English DT Article; Proceedings Paper CT Receptor 2000 Symposium CY APR, 2000 CL LA JOLLA, CALIFORNIA DE muscarinic receptor; M2 subtype; F-18; RS[I-123]IQNB; [F-18]FPTZTP ID IODINE-125-LABELED 1-AZABICYCLO<2.2.2>OCT-3-YL ALPHA-HYDROXY-ALPHA-(1-IODO-1-PRO; POSITRON EMISSION TOMOGRAPHY; IN-VIVO; ALZHEIMERS-DISEASE; RAT-BRAIN; ACETYLCHOLINESTERASE ACTIVITY; M3-MUSCARINIC-RECEPTOR SUBTYPES; AUTORADIOGRAPHIC DISTRIBUTION; 3-QUINUCLIDINYL BENZILATE; COMPUTED-TOMOGRAPHY C1 NIH, Warren G Magnuson Clin Ctr, PET Dept, Bethesda, MD 20814 USA. RP Eckelman, WC (reprint author), NIH, Warren G Magnuson Clin Ctr, PET Dept, Bethesda, MD 20814 USA. NR 41 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 0969-8051 J9 NUCL MED BIOL JI Nucl. Med. Biol. PD JUL PY 2001 VL 28 IS 5 BP 485 EP 491 DI 10.1016/S0969-8051(01)00217-7 PG 7 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 447AT UT WOS:000169546900003 PM 11516692 ER PT J AU Lederman, RJ Raylman, RR Fisher, SJ Kison, PV San, H Nabel, EG Wahl, RL AF Lederman, RJ Raylman, RR Fisher, SJ Kison, PV San, H Nabel, EG Wahl, RL TI Detection of atherosclerosis using a novel positron-sensitive probe and 18-fluorodeoxyglucose (FDG) SO NUCLEAR MEDICINE COMMUNICATIONS LA English DT Article DE atherosclerosis; fludeoxyglucose F18; inflammation; radionuclide imaging; positron ID OPTICAL COHERENCE TOMOGRAPHY; CORONARY-ARTERY DISEASE; IN-VIVO; MYOCARDIAL-INFARCTION; PLAQUE MORPHOLOGY; PROGRESSION; RUPTURE; SITE AB Inflammation contributes to atherosclerotic plaque remodeling, enlargement and rupture. Non-invasive imaging of coronary artery inflammation could help target therapy to 'vulnerable' atheromata, but is limited because of small tissue mass and arterial motion. Local radiopharmaceutical imaging may overcome some of these limitations. We used a positron-sensitive fiberoptic probe, which can distinguish positron emissions from annihilation photons, to identify diseased from healthy endothelium in an atherosclerotic model. New Zealand White rabbits underwent Fogarty-catheter injury of an iliac artery and then were fed a high-fat diet for 3 weeks. Fasted animals received 90-180 MBq of 18-fluorodeoxyglucose (FDG) 2-4 h before sacrifice and harvest of injured and uninjured iliacs. Arteries were incised longitudinally and the probe was placed in contact with the arterial intima. Multiple measurements were obtained along 1 cm artery segments in 60 s intervals, and corrected for F-18 decay and background. Measurements were recorded over 93 injured and normal artery segments in 11 animals. Mean probe Z-scores were 4.8-fold higher (CI 3.4-6.3) over injury atherosclerosis compared with uninjured normal iliac artery segments (P < 0.001). Gamma counting confirmed that injured artery segments accumulated more FDG per gram than did normal segments (0.203%.kg injected dose per gram of tissue versus 0.042, P < 0.001). Non-arterial tissue also accumulated FDG avidly, particularly reticuloendothelial tissues and blood. Delayed sacrifice, 4 h compared with 2 h after animal FDG injection, further reduced blood background counts and improved the signal-to-noise ratio. Histopathology confirmed that injured iliac artery had significantly higher intimal and medial cross-sectional area compared with uninjured artery. Injured artery also had significantly higher macrophage and smooth muscle cell density. Positron-sensitive probe counts correlated with the intima to media ratio (r = 0.63, P = 0.03). Our positron-sensitive probe distinguishes atherosclerotic from healthy artery in a blood-free field. Intravascular study of plaque biology may be feasible using FDG and a positron-sensitive probe. ((C) 2001 Lippincott Williams & Wilkins). C1 Univ Michigan Hlth Syst, Dept Med, Div Cardiol, Ann Arbor, MI USA. RP Lederman, RJ (reprint author), NHLBI, NIH, Bldg 10,Room 8c103, Bethesda, MD 20892 USA. FU NCI NIH HHS [5RO1CA52880] NR 22 TC 109 Z9 112 U1 0 U2 7 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0143-3636 J9 NUCL MED COMMUN JI Nucl. Med. Commun. PD JUL PY 2001 VL 22 IS 7 BP 747 EP 753 DI 10.1097/00006231-200107000-00004 PG 7 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 449TG UT WOS:000169702800004 PM 11453046 ER PT J AU Huang, Y Maraia, RJ AF Huang, Y Maraia, RJ TI Comparison of the RNA polymerase III transcription machinery in Schizosaccharomyces pombe, Saccharomyces cerevisiae and human SO NUCLEIC ACIDS RESEARCH LA English DT Review ID SMALL NUCLEAR-RNA; TATA-BINDING PROTEIN; PROXIMAL SEQUENCE ELEMENT; U6 SNRNA TRANSCRIPTION; IN-VITRO TRANSCRIPTION; TERMINATION FACTOR LA; TFIIIB-DNA COMPLEXES; 2ND LARGEST SUBUNIT; FISSION YEAST; ALPHA-AMANITIN AB Multi-subunit transcription factors (TF) direct RNA polymerase (pol) III to synthesize a variety of essential small transcripts such as tRNAs, 5S rRNA and U6 snRNA, Use by pol III of both TATA-less and TATA-containing promoters, together with progress in the Saccharomyces cerevisiae and human systems towards elucidating the mechanisms of actions of the pol III TFs, provides a paradigm for eukaryotic gene transcription, Human and S. cerevisiae pol III components reveal good general agreement in the arrangement of orthologous TFs that are distributed along tRNA gene control elements, beginning upstream of the transcription initiation site and extending through the 3 ' terminator element, although some TF subunits have diverged beyond recognition. For this review we have surveyed the Schizosaccharomyces pombe database and identified 26 subunits of pol III and associated TFs that would appear to represent the complete core set of the pol III machinery. We also compile data that indicate in vivo expression and/or function of 18 of the fission yeast proteins. A high degree of homology occurs in pol III, TFIIIB, TFIIIA and the three initiation-related subunits of TFIIIC that are associated with the proximal promoter element, while markedly less homology is apparent in the downstream TFIIIC subunits, The idea that the divergence in downstream TFIIIC subunits is associated with differences in pol III termination-related mechanisms that have been noted in the yeast and human systems but not reviewed previously is also considered. C1 NICHHD, Lab Mol Growth Regulat, NIH, Bethesda, MD 20892 USA. RP Maraia, RJ (reprint author), NICHHD, Lab Mol Growth Regulat, NIH, 6 Ctr Dr MSC 2753, Bethesda, MD 20892 USA. NR 181 TC 90 Z9 93 U1 1 U2 2 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0305-1048 J9 NUCLEIC ACIDS RES JI Nucleic Acids Res. PD JUL 1 PY 2001 VL 29 IS 13 BP 2675 EP 2690 DI 10.1093/nar/29.13.2675 PG 16 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 452NM UT WOS:000169864500001 PM 11433012 ER PT J AU Mohaghegh, P Karow, JK Brosh, RM Bohr, VA Hickson, ID AF Mohaghegh, P Karow, JK Brosh, RM Bohr, VA Hickson, ID TI The Bloom's and Werner's syndrome proteins are DNA structure-specific helicases SO NUCLEIC ACIDS RESEARCH LA English DT Article ID SYNDROME GENE-PRODUCT; ROTHMUND-THOMSON-SYNDROME; SINGLE-STRANDED-DNA; ESCHERICHIA-COLI; SACCHAROMYCES-CEREVISIAE; HOLLIDAY JUNCTIONS; RECQ HELICASE; REPLICATION; EXONUCLEASE; BINDING AB BLM and WRN, the products of the Bloom's and Werner's syndrome genes, are members of the RecQ family of DNA helicases, Although both have been shown previously to unwind simple, partial duplex DNA substrates with 3 ' -->5 ' polarity, little is known about the structural features of DNA that determine the substrate specificities of these enzymes. We have compared the substrate specificities of the BLM and WRN proteins using a variety of partial duplex DNA molecules, which are based upon a common core nucleotide sequence. We show that neither BLM nor WRN is capable of unwinding duplex DNA from a blunt-ended terminus or from an internal nick, However, both enzymes efficiently unwind the same blunt-ended duplex containing a centrally located 12 nt single-stranded 'bubble', as well as a synthetic X-structure (a model for the Holiday junction recombination intermediate) in which each 'arm' of the 4-way junction is blunt-ended. Surprisingly, a 3 ' -tailed duplex, a standard substrate for 3 ' -->5 ' helicases, is unwound much less efficiently by BLM and WRN than are the bubble and X-structure substrates, These data show conclusively that a single-stranded 3 ' -tail is not a structural requirement for unwinding of standard B-form DNA by these helicases, BLM and WRN also both unwind a variety of different forms of G-quadruplex DNA, a structure that can form at guanine-rich sequences present at several genomic loci. Our data indicate that BLM and WRN are atypical helicases that are highly DNA structure specific and have similar substrate specificities, We interpret these data in the light of the genomic instability and hyper-recombination characteristics of cells from individuals with Bloom's or Werner's syndrome. C1 Univ Oxford, John Radcliffe Hosp, Imperial Canc Res Fund Labs, Inst Mol Med, Oxford OX3 9DS, England. NIA, Mol Genet Lab, NIH, Baltimore, MD 21224 USA. RP Hickson, ID (reprint author), Univ Oxford, John Radcliffe Hosp, Imperial Canc Res Fund Labs, Inst Mol Med, Oxford OX3 9DS, England. NR 33 TC 373 Z9 382 U1 2 U2 20 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 JUL 1 PY 2001 VL 29 IS 13 BP 2843 EP 2849 DI 10.1093/nar/29.13.2843 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 452NM UT WOS:000169864500020 PM 11433031 ER PT J AU Sigmon, HD Grady, PA AF Sigmon, HD Grady, PA TI Increasing nursing postdoctoral opportunities: National Institute of Nursing Research Spring Science Work Group SO NURSING OUTLOOK LA English DT Article AB On May 1-2, 2000, the National Institute of Nursing Research (NINR) hosted a Spring Science Work Group on Increasing Nursing Postdoctoral Opportunities. The work group met at the National Institutes of Health (NIH) in Bethesda and was chaired by Dr Patricia A. Grady, director, and Dr Hilary D. Sigmon, program director, NINR. The purpose of the meeting was to identify ways to increase postdoctoral opportunities for nurse researchers. The work group considered the gaps and strengths currently present in postdoctoral education and suggested strategies and collaborative approaches for future initiatives. As biomedicine and behavioral science enter the 21st century, nurse scientists have critical roles to play in basic laboratory studies, translational research, clinical investigations, care delivery, and outcome evaluations. To fulfill these roles and to meet the growing need for well-trained nurse scientists in complex, multidisciplinary research, the -pipeline" of postdoctoral nurse researchers must be strengthened. C1 NINR, Off Extramural Programs, Bethesda, MD 20892 USA. RP Sigmon, HD (reprint author), NINR, Off Extramural Programs, Bethesda, MD 20892 USA. NR 0 TC 6 Z9 6 U1 0 U2 1 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0029-6554 J9 NURS OUTLOOK JI Nurs. Outlook PD JUL-AUG PY 2001 VL 49 IS 4 BP 179 EP 181 DI 10.1067/mno.2001.112790 PG 3 WC Nursing SC Nursing GA 467PG UT WOS:000170712500006 PM 11514790 ER PT J AU Bowman, CC Gardner, D AF Bowman, CC Gardner, D TI Building health services research capacity in nursing: Views from members of nursing's leadership SO NURSING OUTLOOK LA English DT Article AB Background: Despite a short history that dates back to the 1960s, health services research (HSR) has become an important force in shaping the delivery system in the current health care environment. Nursing has been noticeably absent from this endeavor, and if it does not increase its presence, it risks missing an important opportunity to influence future directions of health care delivery. Purpose: Our purpose was to evaluate the state of nursing's HSR contributions and to consider ways to increase its capacity in this arena. Method: An interview with four members from nursing's leadership was conducted. Discussion: Nurses can increase the capacity by becoming better collaborators with those who are currently contributing to HSR, both nurses and members of other healthcare disciplines. Also, by reshaping undergraduate and graduate curricula and creating mentorships, nursing can increase its involvement through an informed workforce. Conclusions: Solutions that were offered in this discussion are presented. C1 Vet Affairs San Diego Healthcare Syst, Qual Endhancement Res Initiat, San Diego, CA USA. NIH, Ctr Clin, Bethesda, MD 20892 USA. RP Bowman, CC (reprint author), Vet Affairs San Diego Healthcare Syst, Qual Endhancement Res Initiat, San Diego, CA USA. NR 7 TC 4 Z9 4 U1 0 U2 0 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0029-6554 J9 NURS OUTLOOK JI Nurs. Outlook PD JUL-AUG PY 2001 VL 49 IS 4 BP 187 EP 192 DI 10.1067/mno.2001.116334 PG 6 WC Nursing SC Nursing GA 467PG UT WOS:000170712500008 ER PT J AU Vozarova, B Weyer, C Hanson, K Tataranni, PA Bogardus, C Pratley, RE AF Vozarova, B Weyer, C Hanson, K Tataranni, PA Bogardus, C Pratley, RE TI Circulating interleukin-6 in relation to adiposity, insulin action, and insulin secretion SO OBESITY RESEARCH LA English DT Article DE inflammation; insulin resistance; cytokines; adipose tissue ID CORONARY HEART-DISEASE; DIABETES-MELLITUS; PIMA-INDIANS; RESISTANCE; OBESITY; INFLAMMATION; PATHOGENESIS; DYSFUNCTION; CYTOKINES; TISSUE AB Objective: Plasma concentrations of interleukin-6 (IL-6), a proinflammatory cytokine produced and released in part by adipose tissue, are elevated in people with obesity and type 2 diabetes. Because recent studies suggest that markers of inflammation predict the development of type 2 diabetes, we examined whether circulating plasma IL-6 concentrations were related to direct measures of insulin resistance and insulin secretory dysfunction in Pima Indians, a population with high rates of obesity and type 2 diabetes. Research Methods and Procedures: Fasting plasma IL-6 concentrations (enzyme-linked immunosorbent assay), body composition (DXA), insulin action (M; hyperinsulinemic euglycemic clamp), and acute insulin secretory responses to glucose (25 g intravenous glucose tolerance test) were measured in 58 Pima Indians without diabetes (24 women, 34 men). Results: Fasting plasma IL-6 concentrations were positively correlated with percentage of body fat (r = 0.26, p = 0.049) and negatively correlated with M (r = -0.28, p = 0.031), but were not related to acute insulin response (r = 0.13, p = 0.339). After adjusting for percentage of body fat, plasma IL-6 was not related to M (partial r = -0.23, p = 0,089). Discussion: Fasting plasma IL-6 concentrations are positively related to adiposity and negatively related to insulin action in Pima Indians. The relationship between IL-6 and insulin action seems to be mediated through adiposity. C1 NIDDKD, Clin Diabet & Nutr Sect, NIH, Phoenix, AZ 85016 USA. RP Vozarova, B (reprint author), NIDDKD, Clin Diabet & Nutr Sect, NIH, 4212 N 16th St,Room 5-41, Phoenix, AZ 85016 USA. OI de Courten, Barbora/0000-0001-8760-2511 NR 15 TC 336 Z9 354 U1 1 U2 15 PU NORTH AMER ASSOC STUDY OBESITY PI ROCHESTER PA C/O DR MICHAEL JENSEN, MAYO MEDICAL CENTER, MAYO CLIN 200 FIRST ST, SW, ROCHESTER, MN 55905 USA SN 1071-7323 J9 OBES RES JI Obes. Res. PD JUL PY 2001 VL 9 IS 7 BP 414 EP 417 DI 10.1038/oby.2001.54 PG 4 WC Endocrinology & Metabolism; Nutrition & Dietetics SC Endocrinology & Metabolism; Nutrition & Dietetics GA 452LC UT WOS:000169858300006 PM 11445664 ER PT J AU Schildkraut, JM Cooper, GS Halabi, S Calingaert, B Hartge, P Whittemore, AS AF Schildkraut, JM Cooper, GS Halabi, S Calingaert, B Hartge, P Whittemore, AS TI Age at natural menopause and the risk of epithelial ovarian cancer SO OBSTETRICS AND GYNECOLOGY LA English DT Article ID STATES CASE-CONTROL; COLLABORATIVE ANALYSIS; REPRODUCTIVE FACTORS; INCESSANT OVULATION; DETERMINANTS AB Objective: To investigate the relationship between age at natural menopause and risk of developing epithelial ovarian cancer. Methods: Using data from six population-based, case-control studies conducted in the United States, age at natural menopause among 1411 women with epithelial ovarian cancer and 6380 control subjects were analyzed using survival analysis methods, including Kaplan-Meier and proportional hazards models. Subjects ranged from 20 to 81 years of age. Results: The median age at natural menopause was 50 years among cases compared with 51 years among controls, a difference of borderline statistical significance (P = .06), The hazard ratio for the relationship between case-control status and age at natural menopause was 1.09 (95% confidence interval 0.99, 1.20). Controlling for potential confounders including parity, oral contraceptive use, tubal ligation, smoking, and body mass index did not appreciably change this association. There was little evidence of an association between early age at natural menopause and early onset ovarian cancer (diagnosis age under 48 years). Conclusion: We observed a weak association between ovarian cancer risk and age at natural menopause and, among women with early onset disease, there was little evidence to suggest that early menopause is related to ovarian cancer, Thus, there seems little need for increased surveillance or screening for ovarian cancer among women with early natural menopause. (C) 2001 by the American College of Obstetricians and Gynecologists. C1 Duke Comprehens Canc Ctr, Program Prevent Detect & Control Res, Durham, NC 27710 USA. Natl Inst Environm Hlth Sci, Epidemiol Branch, Canc Ctr Biostat,Duke Comprehens Canc Ctr, Durham, NC 27710 USA. NCI, Epidemiol & Biostat Program, Div Canc Epidemiol & Genet, Stanford, CA USA. Stanford Univ, Dept Hlth Res & Policy, Stanford, CA 94305 USA. RP Schildkraut, JM (reprint author), Duke Comprehens Canc Ctr, Program Prevent Detect & Control Res, Room 238,Hanes House,POB 2949, Durham, NC 27710 USA. FU NCI NIH HHS [CA 76016-01A1] NR 28 TC 32 Z9 33 U1 1 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0029-7844 J9 OBSTET GYNECOL JI Obstet. Gynecol. PD JUL PY 2001 VL 98 IS 1 BP 85 EP 90 DI 10.1016/S0029-7844(01)01388-6 PG 6 WC Obstetrics & Gynecology SC Obstetrics & Gynecology GA 447VF UT WOS:000169592600016 PM 11430962 ER PT J AU Romero, R Gomez, R Chaiworapongsa, T Conoscenti, G Kim, JC Kim, YM AF Romero, R Gomez, R Chaiworapongsa, T Conoscenti, G Kim, JC Kim, YM TI The role of infection in preterm labour and delivery SO PAEDIATRIC AND PERINATAL EPIDEMIOLOGY LA English DT Review ID POLYMERASE-CHAIN-REACTION; PLACEBO-CONTROLLED TRIAL; AMNIOTIC-FLUID GLUCOSE; BLOOD-CELL COUNT; LOW-BIRTH-WEIGHT; RANDOMIZED CONTROLLED TRIAL; RAPID DIAGNOSTIC-TESTS; PREMATURE RUPTURE; INTRAAMNIOTIC INFECTION; BACTERIAL VAGINOSIS C1 NICHHD, Perinatol Res Branch, Bethesda, MD 20892 USA. RP Romero, R (reprint author), Wayne State Univ Hutzel Hosp, NICHD, Perinatol Res Branch, 4707 St Antoine Blvd, Detroit, MI 48201 USA. NR 157 TC 129 Z9 135 U1 0 U2 2 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0269-5022 J9 PAEDIATR PERINAT EP JI Paediatr. Perinat. Epidemiol. PD JUL PY 2001 VL 15 SU 2 BP 41 EP 56 DI 10.1046/j.1365-3016.2001.00007.x PG 16 WC Public, Environmental & Occupational Health; Obstetrics & Gynecology; Pediatrics SC Public, Environmental & Occupational Health; Obstetrics & Gynecology; Pediatrics GA 463PE UT WOS:000170483900006 PM 11520399 ER PT J AU Buekens, P Klebanoff, M AF Buekens, P Klebanoff, M TI Preterm birth research: from disillusion to the search for new mechanisms SO PAEDIATRIC AND PERINATAL EPIDEMIOLOGY LA English DT Editorial Material ID DETERMINANTS; PREGNANCY; DELIVERY; WEIGHT AB No intervention has been shown to decrease the rate of preterm birth. There was thus a need for a new research agenda. The new emphasis is on social and biological mechanisms, including the impact on stress of racism and poverty, and gene-environment interactions. New markers are also under study, and pertain mostly to erection and inflammation. The impact on preterm birth of broad contextual factors, such as universal social protection, will need to be explored further. The recent trends toward increased rates of preterm births deserve much attention. New policies and interventions to decrease medically indicated preterm births should be urgently developed and evaluated. The failure to prevent preterm deliveries has been so disappointing that there is a risk that high rates of preterm births will be seen as unavoidable. The research programme launched by March of Dimes is a timely effort to foster new enthusiasm, to test new ideas and to generate new hypotheses. C1 Univ N Carolina, Sch Publ Hlth, Dept Maternal & Child Hlth, Chapel Hill, NC 27599 USA. NICHHD, Div Epidemiol Stat & Prevent Res, NIH, Bethesda, MD 20892 USA. RP Buekens, P (reprint author), Univ N Carolina, Sch Publ Hlth, Dept Maternal & Child Hlth, Rosenau Hall,CB 7400, Chapel Hill, NC 27599 USA. NR 17 TC 24 Z9 24 U1 1 U2 1 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0269-5022 J9 PAEDIATR PERINAT EP JI Paediatr. Perinat. Epidemiol. PD JUL PY 2001 VL 15 SU 2 BP 159 EP 161 DI 10.1046/j.1365-3016.2001.00015.x PG 3 WC Public, Environmental & Occupational Health; Obstetrics & Gynecology; Pediatrics SC Public, Environmental & Occupational Health; Obstetrics & Gynecology; Pediatrics GA 463PE UT WOS:000170483900014 PM 11520407 ER PT J AU Zhang, J Troendle, JF Levine, RJ AF Zhang, J Troendle, JF Levine, RJ TI Risks of hypertensive disorders in the second pregnancy SO PAEDIATRIC AND PERINATAL EPIDEMIOLOGY LA English DT Article ID SEVERE PREECLAMPSIA; WOMEN; ECLAMPSIA; FREQUENCY; PROGNOSIS; FETAL AB This study examined the incidence of and risk factors for recurrent and newly developed hypertensive disorders in the second pregnancy. We analysed data on 1641 women who had both the first and second pregnancies in the Collaborative Perinatal Project, a large prospective cohort study at 12 hospitals in the US. Nineteen per cent [95% CI 14%, 24%] of women who had gestational hypertension in the first pregnancy, 32% [95% CI 17%, 48%] of those with pre-eclampsia and 46% [95% CI 32%, 60%] of patients with gestational hypertension or pre-eclampsia superimposed on chronic hypertension, had recurrent hypertensive disorders in the second pregnancy. Risk factors for recurrence included history of chronic hypertension and thromboembolism, early onset of hypertension in the first pregnancy or persistent hypertension after 5 weeks postpartum and high baseline blood pressure in the second pregnancy. Women with a normotensive first pregnancy but a severe small-for-gestational-age birth had twice the risk of developing hypertension in the second pregnancy (RR = 2.1, 95% CI, 1.1, 4.0). In summary, hypertensive disorders have a 20-50% recurrence rate in the second pregnancy. The earlier the onset of hypertension in the first pregnancy, the higher the overall recurrence rate. Intrauterine growth restriction of the first birth is an independent risk factor for hypertension in the second pregnancy. C1 NICHHD, Div Epidemiol Stat & Prevent Res, NIH, Bethesda, MD 20892 USA. RP Zhang, J (reprint author), NICHHD, Div Epidemiol Stat & Prevent Res, NIH, Bldg 6100,Room 7B03, Bethesda, MD 20892 USA. NR 16 TC 33 Z9 36 U1 0 U2 1 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0269-5022 J9 PAEDIATR PERINAT EP JI Paediatr. Perinat. Epidemiol. PD JUL PY 2001 VL 15 IS 3 BP 226 EP 231 DI 10.1046/j.1365-3016.2001.00347.x PG 6 WC Public, Environmental & Occupational Health; Obstetrics & Gynecology; Pediatrics SC Public, Environmental & Occupational Health; Obstetrics & Gynecology; Pediatrics GA 456VC UT WOS:000170102100005 PM 11489149 ER PT J AU Wyszynski, DF Diehl, SR AF Wyszynski, DF Diehl, SR TI The mother-only method (MOM) to detect maternal gene-environment interactions SO PAEDIATRIC AND PERINATAL EPIDEMIOLOGY LA English DT Article C1 Boston Univ, Sch Med, Genet Program, Boston, MA 02118 USA. Boston Univ, Sch Publ Hlth, Boston, MA 02118 USA. Natl Inst Dent & Craniofacial Res, NIH, Bethesda, MD USA. RP Wyszynski, DF (reprint author), Boston Univ, Sch Med, Genet Program, 715 Albany St,L320, Boston, MA 02118 USA. NR 3 TC 1 Z9 1 U1 0 U2 0 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0269-5022 J9 PAEDIATR PERINAT EP JI Paediatr. Perinat. Epidemiol. PD JUL PY 2001 VL 15 IS 3 BP 317 EP 318 DI 10.1046/j.1365-3016.2001.00362.x PG 2 WC Public, Environmental & Occupational Health; Obstetrics & Gynecology; Pediatrics SC Public, Environmental & Occupational Health; Obstetrics & Gynecology; Pediatrics GA 456VC UT WOS:000170102100019 PM 11489163 ER PT J AU Nutman, TB Kumaraswami, V AF Nutman, TB Kumaraswami, V TI Regulation of the immune response in lymphatic filariasis: perspectives on acute and chronic infection with Wuchereria bancrofti in South India SO PARASITE IMMUNOLOGY LA English DT Article DE lymphatic filariasis; Wuchereria bancrofti; tolerance; circulating filarial antigen; cytokine; tropical pulmonary eosinophilia ID TROPICAL PULMONARY EOSINOPHILIA; ONCHOCERCA-VOLVULUS INFECTION; PARASITE-SPECIFIC ANERGY; GROWTH-FACTOR-BETA; T-CELL RESPONSES; BRUGIA-MALAYI; INTERLEUKIN-10 PRODUCTION; ALLERGIC SENSITIZATION; CLINICAL STATUS; LYMPHOCYTE-T AB Delineating the immune responses in lymphatic filariasis has been complicated not only by the rapidly expanding knowledge of new immunological mediators and effortors, but also by new methodologies (in particular, circulating filarial antigen detection) for defining and categorizing filarial-infected individuals. By using assays for circulating antigen in the sera collected as part of the many immunological studies performed on individuals in a Wuchereria bancrofti-endemic region of South India, we have attempted to explore the influence of patency on the antigen-driven proliferative and cytokine responses seen in peripheral blood mononuclear cells of individuals with varying clinical manifestations of lymphatic filarial infection. Moreover, we have provided perspectives on the differences between acute and chronic infection with W. bancrofti and suggested mechanisms that may underly the modulation of the immune response as patency occurs. C1 NIAID, Helminth Immunol Sect, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. TB Res Ctr, Chennai, India. RP Nutman, TB (reprint author), NIAID, Helminth Immunol Sect, Parasit Dis Lab, NIH, Bethesda, MD 20892 USA. NR 90 TC 66 Z9 67 U1 0 U2 5 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0141-9838 J9 PARASITE IMMUNOL JI Parasite Immunol. PD JUL PY 2001 VL 23 IS 7 BP 389 EP 399 DI 10.1046/j.1365-3024.2001.00399.x PG 11 WC Immunology; Parasitology SC Immunology; Parasitology GA 456DR UT WOS:000170068300007 PM 11472558 ER PT J AU Min, KT AF Min, KT TI Drosophila as a model to study human brain degenerative diseases SO PARKINSONISM & RELATED DISORDERS LA English DT Article DE adrenoleukodystrophy; bubblegum; Creutzfeldt Jakob disease; eggroll; spongecake; Tay-Sachs disease ID X-LINKED ADRENOLEUKODYSTROPHY; FATTY-ACID METABOLISM; CHILDHOOD ADRENOLEUKODYSTROPHY; DEFECTIVE GLIA; MICE; GENE; PROTEIN; ADRENOMYELONEUROPATHY; IDENTIFICATION; NEUROPATHOLOGY AB Drosophila has been an ideal system in which to identify molecules and define pathways involved in development, in part because of the powerful genetic approaches that are possible. Many of the molecules and pathways important in development in Drosophila are evolutionarily conserved between fly and human. With its highly evolved nervous system, amenability to genetic analysis, and the full genomic sequence available, Drosophila is a valuable tool for investigating and understanding the molecular mechanisms of neurodegenerative diseases. In order to have neurodegenerative Drosophila mutants, I screened EMS treated X chromosomes and P-element inserted 2nd and 3rd chromosomes in Drosophila for reduced life span and neurodegeneration. Twenty-one neurodegenerative mutants including bubblegum, spongecake, and eggroll were isolated and were named by virtue of their brain lesions. Each mutant has distinct pattern of degeneration in specific regions of the brain. Degeneration occurs in lamina and retina region in bubblegum. In spongecake vacuolization can only be seen in the optic lobe, especially in the medulla region. Multilamellated inclusions are wide-spread in the brain of eggroll. It showed not only do the pathologies iin fly brains resemble that of human diseases including Creutzfeldt Jakob disease, Tach-Sachs and Niemann-Pick disease, but the gene involved in the pathological pathway in the bubblegum mutant also functions as in the human adrenoleukodystrophy. Published by Elsevier Science Ltd. C1 NINDS, Neurogenet Branch, NIH, Bethesda, MD 20892 USA. RP Min, KT (reprint author), NINDS, Neurogenet Branch, NIH, Bldg 10-3B14,MSC1250,Room 3B12, Bethesda, MD 20892 USA. OI min, kyung-tai/0000-0003-0983-4258 NR 38 TC 5 Z9 5 U1 0 U2 4 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 1353-8020 J9 PARKINSONISM RELAT D JI Parkinsonism Relat. Disord. PD JUL PY 2001 VL 7 IS 3 BP 165 EP 169 DI 10.1016/S1353-8020(00)00053-5 PG 5 WC Clinical Neurology SC Neurosciences & Neurology GA 436DL UT WOS:000168921200002 ER PT J AU Isacson, O Costantini, L Schumacher, JM Cicchetti, F Chung, S Kim, SK AF Isacson, O Costantini, L Schumacher, JM Cicchetti, F Chung, S Kim, SK TI Cell implantation therapies for Parkinson's disease using neural stem, transgenic or xenogeneic donor cells SO PARKINSONISM & RELATED DISORDERS LA English DT Article DE dopamine; fetal cell; Parkinson's disease; regeneration; stem cells; transplantation ID SUBSTANTIA NIGRA TRANSPLANTS; VENTRAL MESENCEPHALIC GRAFTS; EMBRYONIC DOPAMINE NEURONS; FIBROBLAST GROWTH-FACTOR; CENTRAL-NERVOUS-SYSTEM; ADULT-RAT STRIATUM; LONG-TERM SURVIVAL; INTRASTRIATAL TRANSPLANTS; DENERVATED STRIATUM; FUNCTIONAL RECOVERY AB A new therapeutic neurological and neurosurgical methodology involves cell implantation into the living brain in order to replace intrinsic neuronal systems, that do not spontaneously regenerate after injury, such as the dopaminergic (DA) system affected in Parkinson's disease (PD) and aging. Current clinical data indicate proof of principle for this cell implantation therapy for PD. Furthermore, the disease process does not appear to negatively affect the transplanted cells, although the patient's endogenous DA system degeneration continues. However, the optimal cells for replacement, such as highly specialized human fetal dopaminergic cells capable of repairing an entire degenerated nigrostriatal system, cannot be reliably obtained or generated in sufficient numbers for a standardized medically effective intervention. Xenogeneic and transgenic cell sources of analogous DA cells have shown great utility in animal models and some promise in early pilot studies in PD patients. The cell implantation treatment discipline, using cell fate committed fetal allo- or xenogeneic dopamine neurons and glia, is currently complemented by research on potential stem cell derived DA neurons. Understanding the cell biological principles and developing methodology necessary to generate functional DA progenitors is currently our focus for obtaining DA cells in sufficient quantities for the unmet cell transplantation need for patients with PD and related disorders. (C) 2001 Elsevier Science Ltd. All rights reserved. C1 Harvard Univ, McLean Hosp, Sch Med, Neuroregenerat Lab, Belmont, MA 02478 USA. Harvard Univ, McLean Hosp, Sch Med, Mol Neurobiol Lab, Belmont, MA 02478 USA. Neuroregenerat Lab, Sarasota, FL 34230 USA. Sarasota Mem Hosp, Sarasota, FL 34230 USA. McLean Hosp, NIH, Udall Parkinsons Dis Res Ctr Excellence, Belmont, MA 02478 USA. RP Isacson, O (reprint author), Harvard Univ, McLean Hosp, Sch Med, Neuroregenerat Lab, Belmont, MA 02478 USA. NR 71 TC 21 Z9 25 U1 1 U2 3 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 1353-8020 J9 PARKINSONISM RELAT D JI Parkinsonism Relat. Disord. PD JUL PY 2001 VL 7 IS 3 BP 205 EP 212 DI 10.1016/S1353-8020(00)00059-6 PG 8 WC Clinical Neurology SC Neurosciences & Neurology GA 436DL UT WOS:000168921200008 ER PT J AU Ferre, S Popoli, P Gimenez-Llort, L Rimondini, R Muller, CE Stromberg, I Ogren, O Fuxe, K AF Ferre, S Popoli, P Gimenez-Llort, L Rimondini, R Muller, CE Stromberg, I Ogren, O Fuxe, K TI Adenosine/dopamine interaction: implications for the treatment of Parkinson's disease SO PARKINSONISM & RELATED DISORDERS LA English DT Article DE adenosine receptors; dopamine receptors; receptor/receptor interaction; Parkinson's disease; sensitization; tolerance ID DOPAMINE D-2 RECEPTORS; C-FOS EXPRESSION; COTRANSFECTED FIBROBLAST CELLS; INDUCED ROTATIONAL BEHAVIOR; A(2A) RECEPTOR; RAT STRIATUM; 6-HYDROXYDOPAMINE-LESIONED RATS; TURNING BEHAVIOR; A(1) RECEPTOR; MESSENGER-RNA AB Evidence for a role of dopaminergic neurotransmission in the motor effects of adenosine antagonists, such as caffeine, is reviewed, based on the existence of specific antagonistic interactions between specific subtypes of adenosine and dopamine receptors in the striatum. Both adenosine A(1) and adenosine A(2A) receptor antagonists induce motor activation in rodents. At least a certain degree of dopaminergic activity is required to obtain adenosine antagonist-induced motor activation, with adenosine A(1) antagonists being the most sensitive and non-selective adenosine antagonists the most resistant to striatal dopamine depletion. When considering long-term treatment with adenosine antagonists concomitant administration of dopamine agonists might be required in order to obtain strong motor effects (cross-sensitization) and to avoid the development of telerance. (C) 2001 Elsevier Science Ltd. All rights reserved. C1 Natl Inst Drug Abuse, IRP, NIH, Baltimore, MD 21224 USA. Ist Super Sanita, Dept Pharmacol, I-00161 Rome, Italy. Karolinska Inst, Dept Neurosci, S-17177 Stockholm, Sweden. Univ Wurzburg, Dept Pharmaceut Chem, Inst Pharm & Food Chem, D-97074 Wurzburg, Germany. RP Ferre, S (reprint author), Natl Inst Drug Abuse, IRP, NIH, Baltimore, MD 21224 USA. RI Popoli, Patrizia/B-5397-2008; Rimondini, Roberto/B-2500-2010; Ferre, Sergi/K-6115-2014; Muller, Christa/C-7748-2014; OI Rimondini, Roberto/0000-0003-4099-513X; Ferre, Sergi/0000-0002-1747-1779; Muller, Christa/0000-0002-0013-6624; Ogren, Sven Ove/0000-0003-2573-5276; Fuxe, Kjell/0000-0001-8491-4288 NR 66 TC 89 Z9 90 U1 0 U2 3 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 1353-8020 J9 PARKINSONISM RELAT D JI Parkinsonism Relat. Disord. PD JUL PY 2001 VL 7 IS 3 BP 235 EP 241 DI 10.1016/S1353-8020(00)00063-8 PG 7 WC Clinical Neurology SC Neurosciences & Neurology GA 436DL UT WOS:000168921200012 ER PT J AU Berzofsky, JA AF Berzofsky, JA TI Design of engineered vaccines for systemic and mucosal immunity to HIV SO PATHOLOGIE BIOLOGIE LA English DT Article DE HIV; mucosal immunity; peptides; T lymphocytes; vaccine AB To design vaccines for viruses such as HIV that do not elicit sufficient protective immunity, we first constructed cluster vaccines containing T helper, CTL and neutralizing antibody epitopes. For second generation vaccines, we increased responses by enhancing binding to Major Histocompatibility molecules or by incorporating cytokines. We found that high avidity CTL induce better viral clearance. We also induced anti-HIV mucosal T cell immunity by intrarectal administration. Such approaches may improve classic attenuated or killed pathogen vaccines. (C) 2001 Editions scientifiques et medicales Elsevier SAS. C1 NCI, Mol Immunogenet & Vaccine Res Sect, Metab Branch, NIH, Bethesda, MD 20892 USA. RP Berzofsky, JA (reprint author), NCI, Mol Immunogenet & Vaccine Res Sect, Metab Branch, NIH, Bethesda, MD 20892 USA. NR 1 TC 4 Z9 4 U1 0 U2 1 PU EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER PI PARIS CEDEX 15 PA 23 RUE LINOIS, 75724 PARIS CEDEX 15, FRANCE SN 0369-8114 J9 PATHOL BIOL JI Pathol. Biol. PD JUL PY 2001 VL 49 IS 6 BP 466 EP 467 DI 10.1016/S0369-8114(01)00167-5 PG 2 WC Pathology SC Pathology GA 452JU UT WOS:000169855200007 PM 11484606 ER PT J AU Sokol, GM Fineberg, NS Wright, LL Ehrenkranz, RA AF Sokol, GM Fineberg, NS Wright, LL Ehrenkranz, RA TI Changes in arterial oxygen tension when weaning neonates from inhaled nitric oxide SO PEDIATRIC PULMONOLOGY LA English DT Article; Proceedings Paper CT Annual Meeting of the Pediatric-Academic-Society/Ambulatory-Pediatric-Association CY MAY 01-05, 1998 CL NEW ORLEANS, LOUISIANA SP Pediat Acad Soc, Ambulatory Pediat Assoc DE neonatal hypoxic respiratory failure; pulmonary hypertension; neonatology; nitric oxide; pulmonary circulation; meconium aspiration; pneumonia; sepsis; respiratory distress syndrome; pulmonary hypoplasia; extracorporeal membrane oxygenation; surfactant ID PERSISTENT PULMONARY-HYPERTENSION; DOSE-RESPONSE; RESPIRATORY-FAILURE; NEWBORN; MULTICENTER; INFANTS; THERAPY AB We set out to evaluate changes in arterial oxygen tension (PaO2) when weaning neonates from inhaled nitric oxide (INO). We reviewed the records of 505 prospectively collected INO weaning attempts on 84 neonates with hypoxic respiratory failure. PaO2 values before and 30 min after weaning attempts were recorded. Relationships between change in PaO2 and decreases in INO concentrations were investigated using regression analysis and ANOVA. PaO2 decreased (-18.7 +/- 1.8 torr; P < 0.001) when weaning INO. A stepwise decline in PaO2 was observed weaning INO from 40 ppm. The greatest decline occurred when INO was discontinued (-42.1 +/- 4.1 torr). Forward: stepwise multiple regression using variables with significant relationships to the decline in PaO2 identified the specific dose reduction (P <0.001), the prewean PaO2 (P < 0.001), and surfactant therapy (P = 0.018) as the variables best describing the change in PaO2 (P = 0.004, r = 0.51). In conclusion, a graded decline in PaO2 occurs when reducing INO. INO should be weaned to less than 1 ppm before discontinuing its use. Prior surfactant treatment appears to enhance the oxygenation reserve when weaning INO. Published 2001 Wiley-Liss, Inc. C1 Indiana Univ, Sch Med, Dept Pediat, Indianapolis, IN 46202 USA. Indiana Univ, Sch Med, Dept Med, Indianapolis, IN 46202 USA. NICHD, Bethesda, MD USA. Yale Univ, Sch Med, Dept Pediat, New Haven, CT 06510 USA. RP Sokol, GM (reprint author), Indiana Univ, Sch Med, Dept Pediat, 699 West Dr,RR-208, Indianapolis, IN 46202 USA. FU NCRR NIH HHS [M-01RR750]; NICHD NIH HHS [U10HD27856, U10HD27871] NR 15 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 8755-6863 J9 PEDIATR PULM JI Pediatr. Pulmonol. PD JUL PY 2001 VL 32 IS 1 BP 14 EP 19 DI 10.1002/ppul.1083 PG 6 WC Pediatrics; Respiratory System SC Pediatrics; Respiratory System GA 448LW UT WOS:000169629200003 PM 11416871 ER PT J AU Hunt, CE AF Hunt, CE TI Familial small upper airways and sleep-disordered breathing: Relationship to idiopathic apparent-life-threatening events - Commentary on the article by Guilleminault et al. on page 14 SO PEDIATRIC RESEARCH LA English DT Editorial Material ID INFANT-DEATH-SYNDROME; RECEPTOR-BINDING; APNEA; MICE; OBSTRUCTION; RECORDINGS; GENETICS; CHILDREN; NUCLEUS; GENES C1 NHLBI, Natl Ctr Sleep Disorders Res, Bethesda, MD 20892 USA. RP Hunt, CE (reprint author), NHLBI, Natl Ctr Sleep Disorders Res, 2 Rockledge Ctr,Room 10038,6701 Rockledge Dr,MSC, Bethesda, MD 20892 USA. NR 36 TC 1 Z9 1 U1 0 U2 0 PU INT PEDIATRIC RESEARCH FOUNDATION, INC PI BALTIMORE PA 351 WEST CAMDEN ST, BALTIMORE, MD 21201-2436 USA SN 0031-3998 J9 PEDIATR RES JI Pediatr. Res. PD JUL PY 2001 VL 50 IS 1 BP 3 EP 5 DI 10.1203/00006450-200107000-00003 PG 3 WC Pediatrics SC Pediatrics GA 445LH UT WOS:000169458600001 PM 11420410 ER PT J AU Clancy, RR McGaurn, SA Goin, JE Hirtz, DG Norwood, WI Gaynor, JW Jacobs, ML Wernovsky, G Mahle, WT Murphy, JD Nicolson, SC Steven, JM Spray, TL AF Clancy, RR McGaurn, SA Goin, JE Hirtz, DG Norwood, WI Gaynor, JW Jacobs, ML Wernovsky, G Mahle, WT Murphy, JD Nicolson, SC Steven, JM Spray, TL TI Allopurinol neurocardiac protection trial in infants undergoing heart surgery using deep hypothermic circulatory arrest SO PEDIATRICS LA English DT Article DE congenital heart defects; hypoplastic left heart syndrome; induced hypothermia; ischemia-reperfusion injury; neuroprotective agents; allopurinol; xanthine oxidase; free radicals; seizures; coma ID FLOW CARDIOPULMONARY BYPASS; RANDOMIZED CONTROLLED TRIAL; NORWOOD-PROCEDURE; XANTHINE-OXIDASE; NEUROLOGIC SEQUELAE; SINGLE VENTRICLE; CARDIAC-SURGERY; CHILDREN; ISCHEMIA; PERFORMANCE AB Objective. This pharmacologic protection trial was conducted to test the hypothesis that allopurinol, a scavenger and inhibitor of oxygen free radical production, could reduce death, seizures, coma, and cardiac events in infants who underwent heart surgery using deep hypothermic circulatory arrest (DHCA). Design. This was a single center, randomized, placebo-controlled, blinded trial of allopurinol in infant heart surgery using DHCA. Enrolled infants were stratified as having hypoplastic left heart syndrome (HLHS) and all other forms of congenital heart disease (non-HLHS). Drug was administered before, during, and after surgery. Adverse events and the clinical efficacy endpoints death, seizures, coma, and cardiac events were monitored until infants were discharged from the intensive care unit or 6 weeks, whichever came first. Results. Between July 1992 and September 1997, 350 infants were enrolled and 348 subsequently randomized. A total of 318 infants (131 HLHS and 187 non-HLHS) underwent heart surgery using DHCA. There was a nonsignificant treatment effect for the primary efficacy endpoint analysis (death, seizures, and coma), which was consistent over the 2 strata. The addition of cardiac events to the primary endpoint resulted in a lack of consistency of treatment effect over strata, with the allopurinol treatment group experiencing fewer events (38% vs 60%) in the entire HLHS stratum, compared with the non-HLHS stratum (30% vs 27%). In HLHS surgical survivors, 40 of 47 (85%) allopurinol-treated infants did not experience any endpoint event, compared with 27 of 49 (55%) controls. There were fewer seizures-only and cardiac-only events in the allopurinol versus placebo groups. Allopurinol did not reduce efficacy endpoint events in non-HLHS infants. Treated and control infants did not differ in adverse events. Conclusions. Allopurinol provided significant neuro-cardiac protection in higher-risk HLHS infants who underwent cardiac surgery using DHCA. No benefits were demonstrated in lower risk, non-HLHS infants, and no significant adverse events were associated with allopurinol treatment. C1 Childrens Hosp Philadelphia, Div Neurol, Philadelphia, PA 19104 USA. Childrens Hosp Philadelphia, Div Cardiothorac Surg, Philadelphia, PA 19104 USA. Childrens Hosp Philadelphia, Div Cardiol, Philadelphia, PA 19104 USA. Childrens Hosp Philadelphia, Div Cardiac Anesthesiol, Philadelphia, PA 19104 USA. Univ Penn, Sch Med, Dept Neurol, Philadelphia, PA 19104 USA. Univ Penn, Sch Med, Dept Pediat, Philadelphia, PA 19104 USA. Univ Penn, Sch Med, Dept Surg, Philadelphia, PA 19104 USA. Univ Penn, Sch Med, Dept Anesthesiol & Crit Care Med, Philadelphia, PA 19104 USA. DataMedix Corp, Media, PA USA. Alfred I duPont Hosp Children, Nemours Cardiac Ctr, Wilmington, DE USA. Thomas Jefferson Univ, Jefferson Med Coll, Dept Surg, Philadelphia, PA 19107 USA. Thomas Jefferson Univ, Jefferson Med Coll, Dept Pediat, Philadelphia, PA 19107 USA. St Christophers Hosp Children, Philadelphia, PA 19133 USA. MCP Hahneman Sch Med, Dept Cardiothorac Surg, Philadelphia, PA USA. NINDS, NIH, Bethesda, MD 20892 USA. RP Clancy, RR (reprint author), Childrens Hosp Philadelphia, Div Neurol, 324 S 34th St, Philadelphia, PA 19104 USA. RI gaynor, James william/E-5194-2013 OI gaynor, James william/0000-0001-7955-5604 FU NCRR NIH HHS [MO1-RR0240]; NINDS NIH HHS [NS-N01-2315] NR 44 TC 69 Z9 69 U1 0 U2 2 PU AMER ACAD PEDIATRICS PI ELK GROVE VILLAGE PA 141 NORTH-WEST POINT BLVD,, ELK GROVE VILLAGE, IL 60007-1098 USA SN 0031-4005 J9 PEDIATRICS JI Pediatrics PD JUL PY 2001 VL 108 IS 1 BP 61 EP 70 DI 10.1542/peds.108.1.61 PG 10 WC Pediatrics SC Pediatrics GA 447KU UT WOS:000169571400029 PM 11433055 ER PT J AU Brenner, RA Trumble, AC Smith, GS Kessler, EP Overpeck, MD AF Brenner, RA Trumble, AC Smith, GS Kessler, EP Overpeck, MD TI Where children drown, United States, 1995 SO PEDIATRICS LA English DT Article DE drowning; child; mortality; epidemiology ID NEAR-DROWNINGS; COUNTY; CALIFORNIA; CHILDHOOD AB Context. Drowning is the second leading cause of unintentional injury death among children ages 1 to 19 years. Details about the specific site of submersion are important for implementation of primary prevention efforts, but are not routinely available from national data. Objectives. To provide national data about the specific sites of drowning among US children and to examine site-specific drowning rates by age, race, and gender. Setting. United States, 1995. Design. Information was abstracted from death certificates for unintentional drownings among children <20 years of age. The site of drowning was coded from free text on death certificates. Proportional distributions of the site of drowning by age, region, gender, and race were examined. Site-specific drowning rates were calculated by age, race, and gender. Denominators were based on US Census Bureau, June 1995 estimates of the US population. Results. For 1995, death certificates were obtained for 1420 unintentional drownings among children <20 years of age, 98% of the number of drowning deaths reported by the National Center for Health Statistics for the same year. Site of drowning was specified on 1308 (92%) death certificates. Fifty-five percent of infant drownings were in bathtubs. Among children between the ages of 1 to 4 years, 56% of drownings were in artificial pools and 26% were in other bodies of freshwater. Among older children, 63% of drownings were in natural bodies of freshwater. Site-specific drowning rates varied by race. Importantly, after the age of 5 years, the risk of drowning in a swimming pool was greater among black males compared with white males with rate ratios of 15.1 (95% confidence interval: 6.7-38.5) among 10- to 14-year-olds and 12.8 (95% confidence interval: 6.5-26.9) among 15- to 19-year-olds. Conclusion. The variety of sites in which children drown, even within specific age groups, emphasizes the need for a multifaceted approach to prevention. Reductions in the relatively high rates of drowning among black adolescent males will require targeted interventions to prevent swimming pool drownings among this group. C1 NICHHD, Div Epidemiol Stat & Prevent Res, Bethesda, MD 20892 USA. Johns Hopkins Univ, Sch Publ Hlth, Baltimore, MD USA. Consumer Prod Safety Commiss, Bethesda, MD USA. Maternal & Child Hlth Bur, Rockville, MD USA. RP Brenner, RA (reprint author), NICHHD, Div Epidemiol Stat & Prevent Res, 6100 Execut Blvd,Room 7B03, Bethesda, MD 20892 USA. OI Smith, Gordon/0000-0002-2911-3071 FU NIAAA NIH HHS [R29AA07700] NR 25 TC 61 Z9 61 U1 0 U2 9 PU AMER ACAD PEDIATRICS PI ELK GROVE VILLAGE PA 141 NORTH-WEST POINT BLVD,, ELK GROVE VILLAGE, IL 60007-1098 USA SN 0031-4005 J9 PEDIATRICS JI Pediatrics PD JUL PY 2001 VL 108 IS 1 BP 85 EP 89 DI 10.1542/peds.108.1.85 PG 5 WC Pediatrics SC Pediatrics GA 447KU UT WOS:000169571400032 PM 11433058 ER PT J AU Ito, T Igarashi, H Pradhan, TK Hou, W Mantey, SA Taylor, JE Murphy, WA Coy, DH Jensen, RT AF Ito, T Igarashi, H Pradhan, TK Hou, W Mantey, SA Taylor, JE Murphy, WA Coy, DH Jensen, RT TI GI side-effects of a possible therapeutic GRF analogue in monkeys are likely due to VIP receptor agonist activity SO PEPTIDES LA English DT Article DE GRF; VIP; receptor agonist; PAC(1); growth hormone release ID HORMONE-RELEASING HORMONE; VASOACTIVE-INTESTINAL-PEPTIDE; PIG PANCREATIC ACINI; CANCER CELL-LINES; CYCLASE-ACTIVATING POLYPEPTIDE; SHORT-BOWEL SYNDROME; GROWTH-FACTOR-II; GUINEA-PIG; ADENYLATE-CYCLASE; MOLECULAR-CLONING AB Growth hormone (GH) is used or is being evaluated fbr efficacy in treatment of short stature, aspects of aging, cardiac disorders, Crohn's disease, and short bowel syndrome. Therefore, we synthesized several stable growth hormone-releasing factor (GRF) analogues that could be therapeutically useful. One potent analog, [D-Ala(2),Aib(8,18)Ala(9, 15, 16, 22, 24-26.)Gab(27)]hGRF(1-27)NH2 (GRF-6), with prolonged infusion caused severe diarrhea in monkeys; however, it had no side-effects in rats. Because GRF has similarity to VIP/PACAP and VIPomas cause diarrhea, this study investigated the ability of this and other CRF analogues to interact with the VIP/PACAP receptors. Rat VPAC(1)-R (rVPAC(1)-R), human VPAC(1)-R (hVPAC(1)-R), rVPAC(2)-R and hVPAC(2)-R stably transfected CHO and PANC 1 cells were made and T47D breast cancer cells containing native human VPAC(1)-R and AR4-2J cells containing PAC(1)-R were used. hGRF(1-29)NH2 had low affinity for both rVPAC(1)-R and rVPAC(2)-R while VIP had a high affinity for both receptors. GRF-6 had a low affinity for both rVPAC(1)-R and rVPAC(2)-R and very low affinity for the rPAC(1)-R. VIP had a high affinity, whereas hGRF(1-29)NH, had a low affinity for both hVPAC(1)-R and hVPAC(2)-R. In contrast GRF-6, while having a low affinity for hVPAC(2)-R, had relatively higher affinity for the hVPAC(1)-R. In guinea pig pancreatic acini, all GRF analogues were full agonists at the VPAC1-R causing enzyme secretion. These results demonstrate that in contrast to native hGRF(1-29)NH2 GRF-6 has a relatively high affinity for the human VPAC(1)-R but not for the human VPAC(2)-R, rat VPAC(1)-R, rat VPAC(2)-R or rat PAC(1)-R. These results suggest that the substituted GRF analog, GRF-6, likely causes the diarrheal side-effects in monkeys by interacting with the VPAC(1)-R. Furthermore, they demonstrate significant species differences can exist for possible therapeutic peptide agonists of the VIP/PACAP/GRF receptor family and that it is essential that receptor affinity assessments be performed in human cells or from a closely related species. (C) 2001 Published by Elsevier Science, Inc. C1 NIDDKD, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. Biomeasure Inc, Milford, MA 01757 USA. Tulane Univ, Hlth Sci Ctr, Dept Med, Peptide Res Labs, New Orleans, LA 70112 USA. RP Jensen, RT (reprint author), NIDDKD, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. NR 74 TC 11 Z9 11 U1 1 U2 1 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0196-9781 J9 PEPTIDES JI Peptides PD JUL PY 2001 VL 22 IS 7 BP 1139 EP 1151 DI 10.1016/S0196-9781(01)00436-3 PG 13 WC Biochemistry & Molecular Biology; Endocrinology & Metabolism; Pharmacology & Pharmacy SC Biochemistry & Molecular Biology; Endocrinology & Metabolism; Pharmacology & Pharmacy GA 451KX UT WOS:000169802200018 PM 11445245 ER PT J AU Dai, D Cao, Y Falls, G Levi, PE Hodgson, E Rose, RL AF Dai, D Cao, Y Falls, G Levi, PE Hodgson, E Rose, RL TI Modulation of mouse P450 isoforms CYP1A2, CYP2B10, CYP2E1, and CYP3A by the environmental chemicals mirex, 2,2-bis(p-chlorophenyl)-1,1-dichloroethylene, vinclozolin, and flutamide SO PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY LA English DT Article ID ANDROGEN RECEPTOR ANTAGONIST; RAT-LIVER; INDUCTION; CYTOCHROME-P-450; TESTOSTERONE; MICE; METABOLISM; BIOTRANSFORMATION; HYDROXYLATION; DEALKYLATION AB Several environmental chemicals are disruptive to the reproductive and endocrine systems of many species, including humans. Mechanisms for endocrine disruption are presently under scrutiny. Xenobiotic inducible mammalian cytochrome P450 (CYP) enzymes metabolize a variety of substrates including environmental chemicals, pesticides, and drugs. The metabolism, and thus the effect, of endogenous chemicals including steroid hormones. vitamins, etc. that are transformed by CYP enzymes can be influenced by environmental exposure to CYP-inducing chemicals. This study demonstrated that structurally diverse environmental chemicals including mirex, 2,2-Bis(p-chlorophenyl)-1,1-dichloroethylene (DDE), vinclozolin, and flutamide are capable of inducing several mouse liver CYP isozymes. As demonstrated by Western blotting, mirex induced CYP1A2, 2B10, 2E1, and 3A and vinclozolin induced 1A2 and 2B10. The only isoforms significantly induced by DDE and flutamide were 3A and 1A2, respectively. Since some of these isoforms are known to be involved in metabolism of endogenous hormones, we also studied the effects of these CYP inducers on testosterone metabolism and seminal vesicle weights. Mirex and DDE treatments had profound effects on the metabolism of testosterone, resulting in 2.5- to 3-fold more hydroxylated products than controls. Lesser, but significant, increases in specific metabolites of testosterone were also observed following treatment with vinclozolin and flutamide. Seminal vesicle weights were lower for all treatment groups except DDE. Results of this study demonstrate that, due to their CYP-inducing potential, these chemicals may significantly impact testosterone metabolism and this may be a contributing factor in their antiandrogenic effects. (C) 2001 Academic Press. C1 N Carolina State Univ, Dept Toxicol, Raleigh, NC 27695 USA. NIEHS, Res Triangle Pk, NC 27709 USA. Duke Univ, Dept Radiat Oncol, Durham, NC 27710 USA. RP Rose, RL (reprint author), N Carolina State Univ, Dept Toxicol, Raleigh, NC 27695 USA. NR 46 TC 14 Z9 15 U1 1 U2 3 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0048-3575 J9 PESTIC BIOCHEM PHYS JI Pest. Biochem. Physiol. PD JUL PY 2001 VL 70 IS 3 BP 127 EP 141 DI 10.1006/pest.2001.2551 PG 15 WC Biochemistry & Molecular Biology; Entomology; Physiology SC Biochemistry & Molecular Biology; Entomology; Physiology GA 458CT UT WOS:000170176200001 ER PT J AU Hall, FS Fong, GW Ghaed, S Pert, A AF Hall, FS Fong, GW Ghaed, S Pert, A TI Locomotor-stimulating effects of indirect dopamine agonists are attenuated in Fawn hooded rats independent of postweaning social experience SO PHARMACOLOGY BIOCHEMISTRY AND BEHAVIOR LA English DT Article DE isolation rearing; Fawn hooded rats; dopamine; cocaine; amphetamine ID ALCOHOL-PREFERRING RATS; VENTRAL TEGMENTAL AREA; ELEVATES EXTRACELLULAR DOPAMINE; VOLUNTARY ETHANOL INTAKE; AVOIDING ANA RATS; NUCLEUS-ACCUMBENS; BEHAVIORAL-RESPONSES; ACOUSTIC STARTLE; LABORATORY RATS; WISTAR RATS AB The effects of the indirect dopamine (DA) agonists cocaine and D-amphetamine on locomotor activity were examined in Fawn hooded (FH) rats and Wistar rats. The effect of isolation rearing was also examined to determine if it might have different effects in these two strains. Contrary to previous findings in other rat strains, only small increases in locomotor-stimulating responses to low doses of cocaine were observed in the present study as a result of isolation rearing. However, at higher cocaine doses, locomotor activity was substantially attenuated in FH rats relative to Wistar rats. A similar pattern of effects was observed for amphetamine in FH rats but only at the intermediate dose. The effects of strain and rearing were independent. There was no evidence for interactions between these factors. (C) 2001 Elsevier Science Inc. All rights reserved. C1 NIAAA, DICBR, Clin Studies Lab, Bethesda, MD 20892 USA. Univ Maryland, Dept Psychol, College Pk, MD 20742 USA. NIMH, Biol Psychiat Branch, Bethesda, MD 20892 USA. RP Hall, FS (reprint author), Natl Inst Drug Abuse, IRP, Mol Neurobiol Branch, POB 5180,5500 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Hall, Frank/C-3036-2013 OI Hall, Frank/0000-0002-0822-4063 NR 67 TC 4 Z9 4 U1 0 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0091-3057 J9 PHARMACOL BIOCHEM BE JI Pharmacol. Biochem. Behav. PD JUL-AUG PY 2001 VL 69 IS 3-4 BP 519 EP 526 DI 10.1016/S0091-3057(01)00569-X PG 8 WC Behavioral Sciences; Neurosciences; Pharmacology & Pharmacy SC Behavioral Sciences; Neurosciences & Neurology; Pharmacology & Pharmacy GA 460RP UT WOS:000170322600026 PM 11509212 ER PT J AU Hall, FS Sundstrom, JM Lerner, J Perth, A AF Hall, FS Sundstrom, JM Lerner, J Perth, A TI Enhanced corticosterone release after a modified forced swim test in Fawn hooded rats is independent of rearing experience SO PHARMACOLOGY BIOCHEMISTRY AND BEHAVIOR LA English DT Article DE isolation rearing; forced swim test; Fawn hooded rats; stress; corticosterone; anxiety ID WISTAR RATS; SOCIAL DEPRIVATION; ANIMAL-MODEL; RESPONSES; ETHANOL; ACCUMULATION; HIPPOCAMPUS; IMMOBILITY; DEPRESSION; PREFERENCE AB Recent findings have demonstrated that Fawn hooded (FH/Har) rats exhibit enhanced plasma corticosterone (CORT) responses compared to Wistar rats after exposure to an open field, whereas this effect was not influenced by early social experience. In contrast, it was found that behavior in a modified version of the Porsolt Forced Swim Test (Porsolt FST) was affected by both strain and social experience. An important part of this study included modifications of the Porsolt FST that allowed separation of multiple behavioral endpoints. The present experiment was conducted to determine if FH/Har rats also exhibit enhanced CORT responses after exposure to the modified forced swim test, and whether CORT levels might predict the behavioral response in this context. After the initial exposure in the modified forced swim test FH/Har rats had higher CORT levels than Wistar rats, but this difference was not affected by isolation rearing. However, CORT levels were not correlated with the main behavioral measures assayed in this test. Nonetheless, the data confirm that FH/Har rats have altered HPA axis responses to stressors. (C) 2001 Elsevier Science Inc. All rights reserved. C1 NIAAA, Clin Studies Lab, Bethesda, MD 20892 USA. NIMH, Biol Psychiat Branch, Bethesda, MD 20892 USA. Univ Maryland, Dept Psychol, College Pk, MD 20742 USA. RP Hall, FS (reprint author), NIDA, IRP, Mol Neurobiol Branch, POB 5180,5500 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Hall, Frank/C-3036-2013 OI Hall, Frank/0000-0002-0822-4063 NR 29 TC 24 Z9 24 U1 0 U2 1 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0091-3057 J9 PHARMACOL BIOCHEM BE JI Pharmacol. Biochem. Behav. PD JUL-AUG PY 2001 VL 69 IS 3-4 BP 629 EP 634 DI 10.1016/S0091-3057(01)00556-1 PG 6 WC Behavioral Sciences; Neurosciences; Pharmacology & Pharmacy SC Behavioral Sciences; Neurosciences & Neurology; Pharmacology & Pharmacy GA 460RP UT WOS:000170322600039 PM 11509225 ER PT J AU Eisenhofer, G AF Eisenhofer, G TI The role of neuronal and extraneuronal plasma membrane transporters in the inactivation of peripheral catecholamines SO PHARMACOLOGY & THERAPEUTICS LA English DT Review DE norepinephrine; dopamine; epinephrine; sympathetic nervous system; transporter; uptake ID ORGANIC CATION TRANSPORTER; HUMAN NOREPINEPHRINE TRANSPORTER; HUMAN DOPAMINE TRANSPORTER; IDIOPATHIC DILATED CARDIOMYOPATHY; CARDIAC SYMPATHETIC-NERVES; ANGIOTENSIN-II REGULATION; CONGESTIVE-HEART-FAILURE; RAT ISOLATED HEART; PROTEIN-KINASE-C; I-123 METAIODOBENZYLGUANIDINE UPTAKE AB Catecholamines are translocated across plasma membranes by transporters that belong to two large families with mainly neuronal or extraneuronal locations. In mammals, neuronal uptake of catecholamines involves the dopamine transporter (DAT) at dopaminergic neurons and the norepinephrine transporter (NET) at noradrenergic neurons. Extraneuronal uptake of catecholamines is mediated by organic cation transporters (OCTs), including the classic corticosterone-sensitive extraneuronal monoamine transporter. Catecholamine transporters function as part of uptake and metabolizing systems primarily responsible for inactivation of transmitter released by neurons. Additionally, the neuronal catecholamine transporters, recycle catecholamines for rerelease, thereby reducing requirements for transmitter synthesis. In a broader sense, catecholamine transporters function as part of integrated systems where catecholamine synthesis, release, uptake, and metabolism are regulated in a coordinated fashion in response to the demands placed on the system. Location is also important to function. Neuronal transporters are essential for rapid termination of the signal in neuronal-effector organ transmission, whereas non-neuronal transporters are more important for limiting the spread of the signal and for clearance of catecholamines from the bloodstream. Besides their presynaptic locations, NET and DAT are also present at several extraneuronal locations, including syncytiotrophoblasts of the placenta and endothelial cells of the lung (NET), stomach and pancreas (DAT). The extraneuronal monoamine transporter shows a broad tissue distribution, whereas the other two non-neuronal catecholamine transporters (OCT1 and OCT2) are mainly localized to the liver, kidney, and intestine. Altered function of peripheral catecholamine transporters may be involved in disturbances of the autonomic nervous system, such as occurs in congestive heart failure and hypernoradrenergic hypertension. Peripheral catecholamine transporters provide important targets for clinical imaging of sympathetic nerves and diagnostic localization and treatment of neuroendocrine tumors, such as neuroblastomas and pheochromocytomas. (C) 2001 Elsevier Science Inc. All rights reserved. C1 NINCDS, Clin Neurocardiol Sect, NIH, Bethesda, MD 20892 USA. RP Eisenhofer, G (reprint author), NINCDS, Clin Neurocardiol Sect, NIH, Bldg 10,Room 6N 252,10 Ctr Dr,MSC 1620, Bethesda, MD 20892 USA. EM ge@box-g.nih.gov NR 303 TC 193 Z9 198 U1 0 U2 12 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0163-7258 J9 PHARMACOL THERAPEUT JI Pharmacol. Ther. PD JUL PY 2001 VL 91 IS 1 BP 35 EP 62 DI 10.1016/S0163-7258(01)00144-9 PG 28 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 494NJ UT WOS:000172288500002 PM 11707293 ER PT J AU Weiss, GH Masoliver, J AF Weiss, GH Masoliver, J TI Statistics of dwell times in a reaction with randomly fluctuating rates SO PHYSICA A LA English DT Article DE single-molecule spectroscopy; reaction kinetics; fluctuating rates ID MOLECULE ENZYMATIC DYNAMICS; NON-MARKOVIAN SYSTEMS; SINGLE; FLUORESCENCE; DENSITIES AB Recent single-molecule fluorescence spectroscopy have been analyzed in terms of a reaction with a single fluctuating rare (Shenter. J. Phys. Chem A 103 (1999) 10477). The fluctuations in that analysis are related to the solution of an O-U equation. We propose the use of a simpler type of fluctuation defined in terms of a random telegraph signal. We show that this is equivalent to a four-state Markov model, most of whose properties can be derived exactly in closed form. Published by Elsevier Science B.V. C1 NIH, Div Comp Res & Technol, Math & Stat Comp Lab, Ctr Informat Technol, Bethesda, MD 20892 USA. Univ Barcelona, Dept Fis Fonamental, E-08028 Barcelona, Spain. RP Weiss, GH (reprint author), NIH, Div Comp Res & Technol, Math & Stat Comp Lab, Ctr Informat Technol, Bldg 12A,Room 2007, Bethesda, MD 20892 USA. RI Masoliver, Jaume/F-7198-2016 OI Masoliver, Jaume/0000-0002-5810-879X NR 21 TC 5 Z9 5 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0378-4371 J9 PHYSICA A JI Physica A PD JUL 1 PY 2001 VL 296 IS 1-2 BP 75 EP 82 DI 10.1016/S0378-4371(01)00153-4 PG 8 WC Physics, Multidisciplinary SC Physics GA 447VY UT WOS:000169594200007 ER PT J AU Yen, PM AF Yen, PM TI Physiological and molecular basis of thyroid hormone action SO PHYSIOLOGICAL REVIEWS LA English DT Review ID RETINOID-X-RECEPTOR; RAT GROWTH-HORMONE; BROWN ADIPOSE-TISSUE; LIGAND-BINDING DOMAIN; THYROTROPIN-RELEASING-HORMONE; DOMINANT-NEGATIVE ACTIVITY; MESSENGER-RIBONUCLEIC-ACID; HISTONE DEACETYLASE COMPLEX; SERUM LEPTIN CONCENTRATIONS; TARGET GENE SPECIFICITY AB Thyroid hormones (THs) play critical roles in the differentiation, growth, metabolism, and physiological function of virtually all tissues. TH binds to receptors that are ligand-regulatable transcription factors belonging to the nuclear hormone receptor superfamily. Tremendous progress has been made recently in our understanding of the molecular mechanisms that underlie TH action. In this review, we present the major advances in our knowledge of the molecular mechanisms of TH action and their implications for TH action in specific tissues, resistance to thyroid hormone syndrome, and genetically engineered mouse models. C1 NIDDK, Mol REgulat & Neuroendocrinol Sect, Clin Endocrinol Branch, NIH, Bethesda, MD 20892 USA. RP Yen, PM (reprint author), NIDDK, Mol REgulat & Neuroendocrinol Sect, Clin Endocrinol Branch, NIH, Bethesda, MD 20892 USA. NR 612 TC 1013 Z9 1072 U1 16 U2 96 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0031-9333 J9 PHYSIOL REV JI Physiol. Rev. PD JUL PY 2001 VL 81 IS 3 BP 1097 EP 1142 PG 46 WC Physiology SC Physiology GA 447KQ UT WOS:000169570800005 PM 11427693 ER PT J AU Burbach, JPH Luckman, SM Murphy, D Gainer, H AF Burbach, JPH Luckman, SM Murphy, D Gainer, H TI Gene regulation in the magnocellular hypothalamo-neurohypophysial system SO PHYSIOLOGICAL REVIEWS LA English DT Review ID VASOPRESSIN MESSENGER-RNA; C-FOS EXPRESSION; RAT SUPRAOPTIC NUCLEUS; CORTICOTROPIN-RELEASING-FACTOR; HOMOZYGOUS BRATTLEBORO RAT; NITRIC-OXIDE SYNTHASE; METABOTROPIC GLUTAMATE-RECEPTOR; ALPHA-AMIDATING MONOOXYGENASE; IN-SITU HYBRIDIZATION; IMMEDIATE-EARLY GENES AB The hypothalamo-neurohypophysial system (HNS) is the major peptidergic neurosecretory system through which the brain controls peripheral physiology. The hormones vasopressin and oxytocin released from the HNS at the neurohypophysis serve homeostatic functions of water balance and reproduction. From a physiological viewpoint, the core question on the HNS has always been, "How is the rate of hormone production controlled?" Despite a clear description of the physiology, anatomy, cell biology, and biochemistry of the HNS gained over the last 100 years, this question has remained largely unanswered. However, recently, significant progress has been made through studies of gene identity and gene expression in the magnocellular neurons (MCNs) that constitute the HNS. These are keys to mechanisms and events that exist in the HNS. This review is an inventory of what we know about genes expressed in the HNS, about the regulation of their expression in response to physiological stimuli, and about their function. Genes relevant to the central question include receptors and signal transduction components that receive and process the message that the organism is in demand of a neurohypophysial hormone. The key players in gene regulatory events, the transcription factors, deserve special attention. They do not only control rates of hormone production at the level of the gene, but also determine the molecular make-up of the cell essential for appropriate development and physiological functioning. Finally, the HNS neurons are equipped with a machinery to produce and secrete hormones in a regulated manner. With the availability of several gene transfer approaches applicable to the HNS, it is anticipated that new insights will be obtained on how the HNS is able to respond to the physiological demands for its hormones. C1 Univ Med Ctr, Rudolf Magnus Inst Neurosci, Dept Med Pharmacol, Mol Neurosci Sect, NL-3584 CG Utrecht, Netherlands. Univ Manchester, Sch Biol Sci, Manchester M13 9PL, Lancs, England. Univ Bristol, Dept Med, Mol Neuroendocrinol Res Grp, Bristol, Avon, England. NINDS, Neurochem Lab, NIH, Bethesda, MD 20892 USA. RP Burbach, JPH (reprint author), Univ Med Ctr, Rudolf Magnus Inst Neurosci, Dept Med Pharmacol, Mol Neurosci Sect, Univ Weg 100, NL-3584 CG Utrecht, Netherlands. RI Murphy, David/C-3967-2012; OI Murphy, David/0000-0003-2946-0353; Luckman, Simon/0000-0001-5318-5473 FU Biotechnology and Biological Sciences Research Council [NEU15392] NR 915 TC 203 Z9 204 U1 0 U2 6 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0031-9333 J9 PHYSIOL REV JI Physiol. Rev. PD JUL PY 2001 VL 81 IS 3 BP 1197 EP 1267 PG 71 WC Physiology SC Physiology GA 447KQ UT WOS:000169570800007 PM 11427695 ER PT J AU Topol, IA McGrath, C Chertova, E Dasenbrock, C Lacourse, WR Eissenstat, MA Burt, SK Henderson, LE Casas-Finet, JR AF Topol, IA McGrath, C Chertova, E Dasenbrock, C Lacourse, WR Eissenstat, MA Burt, SK Henderson, LE Casas-Finet, JR TI Experimental determination and calculations of redox potential descriptors of compounds directed against retroviral zinc fingers: Implications for rational drug design SO PROTEIN SCIENCE LA English DT Article DE nucleocapsid; antiretroviral compounds; pulsed polarography; zinc ejection; redox potential; viral inactivation; zinc fingers; density functional theory methods ID TYPE-1 NUCLEOCAPSID PROTEIN; MURINE LEUKEMIA-VIRUS; DIFFERENTIAL PULSE POLAROGRAPHY; PROTON AFFINITIES; RNA ENCAPSIDATION; HIV-1; P7; INACTIVATION; INFECTIVITY; DISULFIRAM AB A diverse set of electrophilic compounds that react with cysteine thiolates in retroviral nucleocapsid (NC) proteins and abolish virus infectivity has been identified. Although different in chemical composition, these compounds are all oxidizing agents that lead to the ejection of Zn(II) ions bound to conserved structural motifs (zinc fingers) present in retroviral NC proteins. The reactivity of a congeneric series of aromatic disulfides toward the NC protein of the human immunodeficiency virus type 1 (HIV-1), NCp7, has been characterized by HPLC separation of starting reagents from reaction products. We calculated the absolute redox potentials of these compounds in the gas phase and in aqueous solvent, using a density functional theory method and a continuum solvation model. Pulsed polarography experiments were performed and showed a direct correlation between calculated and experimentally determined redox propensities. A dependence between protein reactivity and redox potential for a specific compound was shown: Reaction with NCp7 did not take place below a threshold value of redox potential. This relationship permits the distinction between active and nonactive compounds targeted against NCp7, and provides a theoretical basis for a scale of reactivity with retroviral zinc fingers. Our results indicate that electrophilic agents with adequate thiophilicity to react with retroviral NC fingers can now be designed using known or calculated electrochemical properties, This may assist in the design of antiretroviral compounds with greater specificity for NC protein. Such electrophilic agents can be used in retrovirus inactivation with the intent of preparing a whole-killed virus vaccine formulation that exhibits unaffected surface antigenic properties. C1 NCI, SAIC Frederick, AIDS Vaccine Program, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. NCI, SAIC Frederick, Adv Biomed Comp Ctr, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. Univ Maryland Baltimore Cty, Dept Chem & Biochem, Baltimore, MD 21250 USA. NCI, SAIC Frederick, Struct Biochem Program, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RP Henderson, LE (reprint author), NCI, SAIC Frederick, AIDS Vaccine Program, Frederick Canc Res & Dev Ctr, Bldg 535,4th Floor,POB B, Frederick, MD 21702 USA. FU NCI NIH HHS [N01-CO-56000] NR 51 TC 24 Z9 25 U1 1 U2 5 PU COLD SPRING HARBOR LAB PRESS PI PLAINVIEW PA 1 BUNGTOWN RD, PLAINVIEW, NY 11724 USA SN 0961-8368 J9 PROTEIN SCI JI Protein Sci. PD JUL PY 2001 VL 10 IS 7 BP 1434 EP 1445 DI 10.1110/ps.52601 PG 12 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 445KT UT WOS:000169457200016 PM 11420445 ER PT J AU Lee, C Maeng, JS Kocher, JP Lee, B Yu, MH AF Lee, C Maeng, JS Kocher, JP Lee, B Yu, MH TI Cavities of alpha(1)-antitrypsin that play structural and functional roles SO PROTEIN SCIENCE LA English DT Article DE alpha(1)-antitrypsin; cavity-filling mutations; conformational stability; native strain; molecular packing ID REACTIVE CENTER LOOP; AMINO-ACID SUBSTITUTIONS; PROTEIN STRUCTURES; INFLUENZA HEMAGGLUTININ; INHIBITORY MECHANISM; GLOBULAR-PROTEINS; INTERNAL CAVITIES; HYDROPHOBIC CORE; BURIED WATERS; PACKING AB The native form of inhibitory serine protease inhibitors (serpins) is strained, which is critical for their inhibitory activity. Previous studies on stabilizing mutations of alpha (1)-antitrypsin, a prototype of serpins, indicated that cavities provide a structural basis for the native strain of the molecule. We have systematically mapped the cavities of alpha (1)-antitrypsin that play such structural and functional roles by designing cavity-filling mutations at residues that line the walls of the cavities, Results show that energetically unfavorable cavities are distributed throughout the alpha (1)-antitrypsin molecule, and the cavity-filling mutations stabilized the native conformation at 8 out of 10 target sites. The stabilization effect of the individual cavity-filling mutations of alpha (1)-antitrypsin varied (0.2-1.9 kcal/mol for each additional methylene group) and appeared to depend largely on the structural flexibility of the cavity environment. Cavity-filling mutations that decreased inhibitory activity of alpha (1)-antitrypsin were localized in the loop regions that interact with beta -sheet A distal from the reactive center loop. The results are consistent with the notion that beta -sheet A and the structure around it mobilize when alpha (1)-antitrypsin forms a complex with a target protease. C1 Korea Inst Sci & Technol, Prot Strain Res Ctr, Natl Creat Res Initiat, Seoul 130650, South Korea. NCI, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Yu, MH (reprint author), Korea Inst Sci & Technol, Prot Strain Res Ctr, Natl Creat Res Initiat, POB 131, Seoul 130650, South Korea. NR 42 TC 36 Z9 36 U1 1 U2 6 PU COLD SPRING HARBOR LAB PRESS PI PLAINVIEW PA 1 BUNGTOWN RD, PLAINVIEW, NY 11724 USA SN 0961-8368 J9 PROTEIN SCI JI Protein Sci. PD JUL PY 2001 VL 10 IS 7 BP 1446 EP 1453 DI 10.1110/ps.840101 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 445KT UT WOS:000169457200017 PM 11420446 ER PT J AU Miller, M Ginalski, K Lesyng, B Nakaigawa, N Schmidt, L Zbar, B AF Miller, M Ginalski, K Lesyng, B Nakaigawa, N Schmidt, L Zbar, B TI Structural basis of oncogenic activation caused by point mutations in the kinase domain of the MET proto-oncogene: Modeling studies SO PROTEINS-STRUCTURE FUNCTION AND GENETICS LA English DT Article DE MET proto-oncogene; receptor tyrosine kinase; oncogenic mutations; homology modeling; substrate specificity ID RECEPTOR TYROSINE KINASE; PAPILLARY RENAL CARCINOMAS; CRYSTAL-STRUCTURE; C-MET; INSULIN-RECEPTOR; DOCKING SITE; GROWTH; SRC; TRANSFORMATION; GERMLINE AB Missense mutations in the tyrosine kinase domain of the MET proto-oncogene occur in selected cases of papillary renal carcinoma. in biochemical and biological assays, these mutations produced constitutive activation of the MET kinase and led to tumor formation in nude mice. Some mutations caused transformation of NM 3T3 cells. To elucidate the mechanism of ligand-independent MET kinase activation by point mutations, we constructed several 3D models of the wild-type and mutated MET catalytic core domains, Analysis of these structures showed that some mutations (e,g,, V1110I, Y1248H/D/C, M1268T) directly alter contacts between residues from the activation loop in its inhibitory conformation and those from the main body of the catalytic domain; others (e.g., M1149T, L1213V) increase flexibility at the critical paints of the tertiary structure and facilitate subdomain movements, Mutation D1246N plays a role in stabilizing the active form of the enzyme, Mutation M1268T affects the S+1 and S+3 substrate-binding pockets, Models implicate that although these changes do not compromise the affinity toward the C-terminal autophosphorylation site of the MET protein, they allow for binding of the substrate for the c-Abl tyrosine kinase. We provide biochemical data supporting this observation, Mutation L1213V affects the conformation of Tyr1212 in the active form of MET. Several somatic mutations are clustered at the surface of the catalytic domain in close vicinity of the probable location of the MET C-terminal docking site for cytoplasmic effecters, Proteins 2001;44:32-43. (C) 2001 Wiley-Liss, Inc.*. C1 NCI, Macromol Crystallog Lab, Frederick, MD 21702 USA. Univ Warsaw, Inst Expt Phys, Dept Biophys, Warsaw, Poland. Univ Warsaw, Interdisciplinary Ctr Math & Computat Model, Warsaw, Poland. NCI, Immunobiol Lab, Frederick, MD 21701 USA. NCI, Intramural Res Support Program, SAIC Frederick, Frederick, MD 21701 USA. RP Miller, M (reprint author), NCI, Macromol Crystallog Lab, POB B, Frederick, MD 21702 USA. RI Miller, Maria/I-1636-2013 OI Miller, Maria/0000-0003-0252-5348 FU NCI NIH HHS [N01-CO-56000] NR 51 TC 38 Z9 40 U1 0 U2 3 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0887-3585 J9 PROTEINS JI Proteins PD JUL 1 PY 2001 VL 44 IS 1 BP 32 EP 43 DI 10.1002/prot.1069 PG 12 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 438RH UT WOS:000169067000005 PM 11354004 ER PT J AU Senior, C AF Senior, C TI Membership fees SO PSYCHOLOGIST LA English DT Letter C1 NIMH, Sect Cognit Neuropsychol, Lab Brain & Cognit, Bethesda, MD 20892 USA. RP Senior, C (reprint author), NIMH, Sect Cognit Neuropsychol, Lab Brain & Cognit, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU BRITISH PSYCHOLOGICAL SOC PI LEICESTER PA ST ANDREWS HOUSE, 48 PRINCESS RD EAST, LEICESTER LE1 7DR, LEICS, ENGLAND SN 0952-8229 J9 PSYCHOLOGIST JI Psychologist PD JUL PY 2001 VL 14 IS 7 BP 349 EP 349 PG 1 WC Psychology, Multidisciplinary SC Psychology GA 453CN UT WOS:000169899100016 ER PT J AU Champoux, M Hwang, L Lang, O Levine, S AF Champoux, M Hwang, L Lang, O Levine, S TI Feeding demand conditions and plasma cortisol in socially-housed squirrel monkey mother-infant dyads SO PSYCHONEUROENDOCRINOLOGY LA English DT Article DE cortisol; squirrel monkey; stress; feeding; mother-infant; separation ID CEREBROSPINAL-FLUID CONCENTRATIONS; CORTICOTROPIN-RELEASING FACTOR; RHESUS-MONKEYS; BEHAVIORAL-RESPONSES; CAPTIVE CHIMPANZEES; SAIMIRI-SCIUREUS; ENRICHMENT; SEPARATION; PRIMATES; STRESS AB Previous studies have demonstrated that experimentally altering the accessibility and availability of food can have profound impact on behavior and adrenocortical activity in nonhuman primate species. In this study, groups of mother-infant squirrel monkey dyads were housed in either high demand (HFD: 120% normal daily food intake provided), low demand (LFD: 600% normal daily food intake provided) or variable demand (VFD: alternating two-week blocks of low demand and high demand) conditions for 12 weeks. During the 12-week experimental foraging phase, animals in the HFD group exhibited prolonged and consistent cortisol elevations. The cortisol levels in the VFD group reflected the ambient demand condition, with higher levels exhibited during the high demand phases of the study, and lower values when the low demand condition was in effect. Overall, mothers were more affected by the experimental manipulation than were infants. The experimental condition did not affect the infants' response to a 24-h separation from their mothers. A suppression of cortisol levels, particularly in the HFD group, was observed upon resumption of ad-libitum feeding. (C) 2001 Elsevier Science Ltd. All rights reserved. C1 Stanford Univ, Sch Med, Dept Psychiat & Behav Sci, Stanford, CA 94305 USA. RP Champoux, M (reprint author), NIH, Anim Ctr, POB 529, Poolesville, MD 20837 USA. FU NICHD NIH HHS [HD02881]; NIMH NIH HHS [MH15147-10, MH09912-01] NR 43 TC 11 Z9 11 U1 2 U2 4 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0306-4530 J9 PSYCHONEUROENDOCRINO JI Psychoneuroendocrinology PD JUL PY 2001 VL 26 IS 5 BP 461 EP 477 DI 10.1016/S0306-4530(01)00006-3 PG 17 WC Endocrinology & Metabolism; Neurosciences; Psychiatry SC Endocrinology & Metabolism; Neurosciences & Neurology; Psychiatry GA 445MC UT WOS:000169460400002 PM 11337131 ER PT J AU Sabatinelli, D Bradley, MM Lang, PJ AF Sabatinelli, D Bradley, MM Lang, PJ TI Affective startle modulation in anticipation and perception SO PSYCHOPHYSIOLOGY LA English DT Article DE emotion; anticipation; startle reflex; affective pictures ID BLINK REFLEX; FEAR AB Startle modulation was investigated as participants first anticipated and then viewed affective pictures in order to determine whether affective modulation of the startle reflex is similar in these different task contexts. During a 6-s anticipation period, a neutral light cue signaled whether the upcoming picture would portray snakes, erotica, or household objects; at the end of the anticipatory period, a picture in the signaled category was viewed for 6 s. Male participants highly fearful of snakes were recruited to maximize emotional arousal during anticipation and perception. Results indicated that the startle reflex was potentiated when anticipating either unpleasant (phobic) or pleasant (erotic) pictures, compared to neutral stimuli, whereas during perception, reflexes were potentiated when viewing unpleasant stimuli, and reduced when viewing pleasant pictures. The startle reflex is modulated by hedonic valence in picture perception, and by emotional arousal in a task context involving picture anticipation. C1 Univ Florida, Natl Inst Mental Hlth, Ctr Study Emot & Attent, Gainesville, FL 32610 USA. RP Bradley, MM (reprint author), Univ Florida, Natl Inst Mental Hlth, Ctr Study Emot & Attent, Box 100165 HSC, Gainesville, FL 32610 USA. RI Frank, David/E-8213-2012; OI sabatinelli, dean/0000-0001-7409-8504 FU NIMH NIH HHS [MH37757, MH43975, P50-MH52384] NR 20 TC 58 Z9 60 U1 3 U2 16 PU CAMBRIDGE UNIV PRESS PI PORT CHESTER PA 110 MIDLAND AVE, PORT CHESTER, NY 10573-9863 USA SN 0048-5772 J9 PSYCHOPHYSIOLOGY JI Psychophysiology PD JUL PY 2001 VL 38 IS 4 BP 719 EP 722 PG 4 WC Psychology, Biological; Neurosciences; Physiology; Psychology; Psychology, Experimental SC Psychology; Neurosciences & Neurology; Physiology GA 446NM UT WOS:000169519400014 PM 11446586 ER PT J AU Bosworth, HB Bastian, LA Kuchibhatla, MN Steffens, DC McBride, CM Skinner, CS Rimer, BK Siegler, IC AF Bosworth, HB Bastian, LA Kuchibhatla, MN Steffens, DC McBride, CM Skinner, CS Rimer, BK Siegler, IC TI Depressive symptoms, menopausal status, and climacteric symptoms in women at midlife SO PSYCHOSOMATIC MEDICINE LA English DT Article DE menopause; depression; climacteric symptoms ID HORMONE REPLACEMENT THERAPY; POSTMENOPAUSAL WOMEN; ESTROGEN REPLACEMENT; COMMUNITY SAMPLE; MOOD; PREVALENCE; HEALTH; SCALE; PREDICTORS; PROGESTIN AB Objective: Previous studies have found increased rates of depression in women aged 45 to 54 years, but the factors that influence these rates are not understood. It was assessed whether higher rates of depressive symptoms were associated with menopausal status, climacteric symptoms, and use of hormone replacement therapy. Design: Cross-sectional survey. Setting: Community sample. Methods: Data are from 581 women ages 45 to 54 years who were interviewed by telephone between October 1998 and February 1999. Measures: Depression was measured with the abbreviated CES-D, a depressive symptoms screening measure. Women's reported perception of menopausal stage, frequency of periods in the preceding 12 months, and history of oophorectomy were used to classify their menopausal status into four categories: (1) no indication of menopause; (2) close to menopause; (3) had begun menopause; and (4) had completed menopause. Results: There were 168 women (28.9%) who reported a high level (greater than or equal to 10) of depressive symptoms when the abbreviated CES-D was used. In a logistic-regression analysis, significant factors associated with increased depressive symptoms included physical inactivity, inadequate income, use of estrogen/progesterone combination, and presence of climacteric symptoms (trouble sleeping, mood swings, or memory problems). Menopausal status was not associated with depressive symptoms. Conclusions: In this sample of women age 45 to 54 years, climacteric symptoms but not menopausal status were associated with higher rates of depressive symptoms. C1 VAMC 152, Hlth Serv Res & Dev, Durham, NC 27707 USA. Duke Univ, Dept Med, Div Gen Internal Med, Durham, NC USA. Duke Univ, Dept Psychiat & Behav Sci, Durham, NC USA. Duke Univ, Ctr Aging & Human Dev, Durham, NC USA. Duke Univ, Canc Prevent Detect & Control Res Program, Duke Comprehens Canc Ctr, Durham, NC USA. Duke Univ, Dept Community & Family Med, Durham, NC USA. NCI, Div Canc Control & Populat Studies, Bethesda, MD 20892 USA. RP Bosworth, HB (reprint author), VAMC 152, Hlth Serv Res & Dev, 508 Fulton St, Durham, NC 27707 USA. FU NCI NIH HHS [5U19-CA72099] NR 46 TC 107 Z9 109 U1 1 U2 3 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0033-3174 J9 PSYCHOSOM MED JI Psychosom. Med. PD JUL-AUG PY 2001 VL 63 IS 4 BP 603 EP 608 PG 6 WC Psychiatry; Psychology; Psychology, Multidisciplinary SC Psychiatry; Psychology GA 457CP UT WOS:000170119800011 PM 11485114 ER PT J AU Allen, J Markovitz, J Jacobs, DR Knox, SS AF Allen, J Markovitz, J Jacobs, DR Knox, SS TI Social support and health behavior in hostile black and white men and women in CARDIA SO PSYCHOSOMATIC MEDICINE LA English DT Article DE hostility; social support; health behavior ID CORONARY HEART-DISEASE; ADOLESCENT CIGARETTE-SMOKING; FACTOR-INTERVENTION-TRIAL; WEIGHT-RELATED ATTITUDES; ALL-CAUSE MORTALITY; PHYSICAL-ACTIVITY; YOUNG-ADULTS; CARDIOVASCULAR-DISEASE; ALCOHOL-CONSUMPTION; AFRICAN-AMERICAN AB Objective: These cross-sectional analyses of the Coronary Artery Risk Development in Young Adults (CARDIA) data were stimulated by previous CARDIA analyses that showed an adverse association between hostility and several health behaviors: physical activity, cigarette smoking, alcohol consumption, and caloric intake, in both black and white men and women, such that the higher the hostility, the worse the health behavior profile. The current study investigated whether high social support was associated with better health behavior than low social support in individuals with high hostility scores. Methods: The subjects were 5115 healthy black and white men and women ranging in age from 18 to 30 years. The hypothesis was that the association between hostility and certain adverse health behaviors would be diminished in the presence of high social support. Race-gender specific median cutpoints of the Cook-Medley Hostility scale and an index of social support defined levels of high and low hostility and social support. Results: After controlling for age and body mass index (BMI), support was positively associated with more exercise in all groups except black women, but when coupled with high hostility, this positive association between support and exercise remained only in men. White women with high support were less often smokers but this association did not hold when examined only in the high-hostile group. Black men and white women with high support in the presence of high hostility consumed more alcohol, but the amount was moderate. Conclusions: We conclude that social support in the presence of high hostility only sometimes reduces the association of hostility to adverse health behaviors and that these effects are complex. Additional research investigating types of social support on health behavior in different race-gender groups is advocated. C1 NHLBI, Div Epidemiol & Clin Applicat, Bethesda, MD 20892 USA. Univ Alabama, Div Prevent Med, Birmingham, AL USA. Univ Minnesota, Div Epidemiol, Minneapolis, MN 55455 USA. RP Knox, SS (reprint author), NHLBI, Div Epidemiol & Clin Applicat, 6701 Rockledge Dr, Bethesda, MD 20892 USA. FU NHLBI NIH HHS [N01-HC-48047, N01-HC-48048, N01-HC-48049, N01-HC-48050, N01-HC-95095] NR 92 TC 29 Z9 31 U1 5 U2 6 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0033-3174 J9 PSYCHOSOM MED JI Psychosom. Med. PD JUL-AUG PY 2001 VL 63 IS 4 BP 609 EP 618 PG 10 WC Psychiatry; Psychology; Psychology, Multidisciplinary SC Psychiatry; Psychology GA 457CP UT WOS:000170119800012 PM 11485115 ER PT J AU Miller, JH Bolger, G Kempner, E AF Miller, JH Bolger, G Kempner, E TI Radiation target analysis of enzymes with stable free radicals SO RADIATION PHYSICS AND CHEMISTRY LA English DT Article; Proceedings Paper CT Pacifichem 2000 Meeting CY DEC 14-19, 2000 CL HONOLULU, HAWAII DE target size; protein function and protein structure; ribonucleotide reductase; galactose oxidase; radiation inactivation; energy transfer ID SIMPLEX VIRUS TYPE-1; COLI RIBONUCLEOTIDE REDUCTASE; ESCHERICHIA-COLI; GALACTOSE-OXIDASE; PROTEIN R2; INACTIVATION; PHENYLALANINE; SEQUENCE; SUBUNIT; SITE AB Irradiation of ribonucleotide reductase yielded a functional target size of the entire tetramer. The free radical-containing R2 subunit target size for activity was much larger than the physical structure while that for loss of structure was similar to the monomer. This indicates that indirect effects of ionizing radiation removed the free radical in the isolated R2 without substantial structural damage. Preliminary results of galactose oxidase indicate a target mass equal to the monomer size. No indirect effects were observed on the free radical which is not accessible to the solvent. Published by Elsevier Science Ltd. C1 NIAMSD, Phys Biol Lab, NIH, Bethesda, MD 20892 USA. Boehringer Ingelheim Canada Ltd, Dept Biol Sci, BioMega Res Div, Laval, PQ H7S 2G5, Canada. RP Kempner, E (reprint author), NIAMSD, Phys Biol Lab, NIH, Bldg 6,Room 140, Bethesda, MD 20892 USA. NR 25 TC 0 Z9 0 U1 0 U2 0 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0969-806X J9 RADIAT PHYS CHEM JI Radiat. Phys. Chem. PD JUL PY 2001 VL 62 IS 1 BP 33 EP 38 DI 10.1016/S0969-806X(01)00419-4 PG 6 WC Chemistry, Physical; Nuclear Science & Technology; Physics, Atomic, Molecular & Chemical SC Chemistry; Nuclear Science & Technology; Physics GA 474BG UT WOS:000171084000006 ER PT J AU Farb, RI McGregor, C Kim, JK Laliberte, M Derbyshire, JA Willinsky, RA Cooper, PW Westman, DG Cheung, G Schwartz, ML Stainsby, JA Wright, GA AF Farb, RI McGregor, C Kim, JK Laliberte, M Derbyshire, JA Willinsky, RA Cooper, PW Westman, DG Cheung, G Schwartz, ML Stainsby, JA Wright, GA TI Intracranial arteriovenous malformations: Real-time auto-triggered elliptic centric-ordered 3D gadolinium-enhanced MR angiography - Initial assessment SO RADIOLOGY LA English DT Article DE arteriovenous malformations; cerebral blood vessels, abnormalities; cerebral blood vessels, MR; magnetic resonance (MR), technology; magnetic resonance (MR), time of flight ID MAGNETIC-RESONANCE ANGIOGRAPHY; CONTRAST; ARTERIES; RADIOSURGERY; ACQUISITION; OPTIMIZATION; CIRCULATION; DATABASE; ARRIVAL AB Auto-triggered elliptic centric-ordered three-dimensional (3D) gadolinium-enhanced magnetic resonance (MR) angiography was compared with 3D multiple overlapping thin-slab acquisition time-of-flight (TOF) MR angiography in the evaluation of intracranial arteriovenous malformations (AVMs) in 10 patients. Intraarterial digital subtraction angiography (DSA) was the reference standard. Gadolinium-enhanced MR angiograms were found to be equivalent to DSA images in AVM component depiction in 70%-90% of cases and were consistently superior to TOF MR angiograms. C1 Univ Toronto, Toronto Western Hosp, Univ Hlth Network, Dept Med Imaging,Div Neuroradiol, Toronto, ON M5T 2S8, Canada. Univ Toronto, Toronto Western Hosp, Univ Hlth Network, Dept Surg,Div Neurosurg, Toronto, ON M5T 2S8, Canada. Univ Toronto, Toronto Western Hosp, Univ Hlth Network, Dept Med Biophys, Toronto, ON M5T 2S8, Canada. NHLBI, Cardiac Energet Lab, NIH, Bethesda, MD 20892 USA. Sunnybrook & Womens Coll, Hlth Sci Ctr, Dept Imaging Res, Toronto, ON, Canada. RP Farb, RI (reprint author), Univ Toronto, Toronto Western Hosp, Univ Hlth Network, Dept Med Imaging,Div Neuroradiol, Fell Pavil 3-404,300 Bathurst St, Toronto, ON M5T 2S8, Canada. NR 31 TC 62 Z9 68 U1 0 U2 1 PU RADIOLOGICAL SOC NORTH AMER PI EASTON PA 20TH AND NORTHAMPTON STS, EASTON, PA 18042 USA SN 0033-8419 J9 RADIOLOGY JI Radiology PD JUL PY 2001 VL 220 IS 1 BP 244 EP 251 PG 8 WC Radiology, Nuclear Medicine & Medical Imaging SC Radiology, Nuclear Medicine & Medical Imaging GA 445QH UT WOS:000169468400039 PM 11426005 ER PT J AU George, JD Price, CJ Marr, MC Myers, CB Jahnke, GD AF George, JD Price, CJ Marr, MC Myers, CB Jahnke, GD TI Developmental toxicity evaluation of 1,2,3,4-butanetetracarboxylic acid in Sprague Dawley (CD (R)) rats SO REPRODUCTIVE TOXICOLOGY LA English DT Article DE 1,2,3,4-butanetetracarboxylic acids developmental toxicity; teratogenicity; rats; morphologic development ID POLYCARBOXYLIC ACIDS; CITRIC-ACID; COTTON; FORMALDEHYDE; BTCA; ADDITIVES; PERFORMANCE; CATALYST; FABRICS; AGENTS AB 1,2,3,4-butanetetracarboxylic acid (BTCA), proposed as a formaldehyde substitute in the treatment of permanent press fabrics, was evaluated for developmental toxicity. Timed-mated CD rats (25 per group) received BTCA 250, 500, or 1000 mg/kg/day or vehicle (deionized/distilled water) by gavage on gestational days (gd) 6 through 19. Maternal feed and water consumption, body weight, and clinical signs were monitored throughout gestation. At termination (gd 20), confirmed-pregnant females (21 to 25 per group) were evaluated for clinical status and gestational outcome; live fetuses were examined for external, visceral, and skeletal malformations. One maternal death, reduced body weight, and reduced weight gain were noted at the high dose; confirmed pregnancy rates were 84 to 100% for each group. There were no treatment-related effects on fetal growth, survival, or morphologic development. The maternal toxicity NOAEL and LOAEL are 500 and 1000 mg/kg/day, respectively. The developmental toxicity NOAEL is 1000 mg/kg/day, and the LOAEL was not established in this study. (C) 2001 Elsevier Science Inc. All rights reserved. C1 Res Triangle Inst, Ctr Life Sci & Toxicol, Res Triangle Pk, NC 27709 USA. Natl Inst Environm Hlth Sci, Natl Toxicol Program, Dev & Reprod Toxicol Grp, Res Triangle Pk, NC 27709 USA. RP George, JD (reprint author), Res Triangle Inst, Ctr Life Sci & Toxicol, POB 12194, Res Triangle Pk, NC 27709 USA. FU NIEHS NIH HHS [N01-ES-65405] NR 57 TC 2 Z9 2 U1 2 U2 6 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0890-6238 J9 REPROD TOXICOL JI Reprod. Toxicol. PD JUL-AUG PY 2001 VL 15 IS 4 BP 413 EP 420 DI 10.1016/S0890-6238(01)00142-3 PG 8 WC Reproductive Biology; Toxicology SC Reproductive Biology; Toxicology GA 462YL UT WOS:000170447900007 PM 11489597 ER PT J AU Matzinger, P AF Matzinger, P TI Introduction to the series SO SCANDINAVIAN JOURNAL OF IMMUNOLOGY LA English DT Editorial Material ID VERSUS-HOST DISEASE C1 NIAID, Cellular & Mol Immunol Lab, Ghost Lab, NIH, Bethesda, MD 20892 USA. RP Matzinger, P (reprint author), NIAID, Cellular & Mol Immunol Lab, Ghost Lab, NIH, 9000 Rockville Pike,Bldg 4,Room 111, Bethesda, MD 20892 USA. NR 7 TC 24 Z9 28 U1 0 U2 2 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0300-9475 J9 SCAND J IMMUNOL JI Scand. J. Immunol. PD JUL-AUG PY 2001 VL 54 IS 1-2 BP 2 EP 3 DI 10.1046/j.1365-3083.2001.00973.x PG 2 WC Immunology SC Immunology GA 452ZT UT WOS:000169890300001 PM 11439141 ER PT J AU Matzinger, P AF Matzinger, P TI Essay 1: The danger model in its historical context SO SCANDINAVIAN JOURNAL OF IMMUNOLOGY LA English DT Editorial Material ID DELTA T-CELLS; SELF-NONSELF DISCRIMINATION; IMMUNOLOGY; TOLERANCE C1 NIAID, NIH, Bethesda, MD 20892 USA. RP Matzinger, P (reprint author), NIAID, NIH, 9000 Rockville Pike,Bldg 4, Bethesda, MD 20892 USA. NR 32 TC 84 Z9 93 U1 0 U2 4 PU BLACKWELL SCIENCE LTD PI OXFORD PA P O BOX 88, OSNEY MEAD, OXFORD OX2 0NE, OXON, ENGLAND SN 0300-9475 J9 SCAND J IMMUNOL JI Scand. J. Immunol. PD JUL-AUG PY 2001 VL 54 IS 1-2 BP 4 EP 9 DI 10.1046/j.1365-3083.2001.00974.x PG 6 WC Immunology SC Immunology GA 452ZT UT WOS:000169890300002 PM 11439142 ER PT J AU Ruffini, PA Kwak, LW AF Ruffini, PA Kwak, LW TI Immunotherapy of multiple myeloma SO SEMINARS IN HEMATOLOGY LA English DT Article ID MAJOR HISTOCOMPATIBILITY COMPLEX; STEM-CELL TRANSPLANTATION; COLONY-STIMULATING FACTOR; CD4(+) T-CELLS; DENDRITIC CELLS; TUMOR-ANTIGEN; SYNDECAN-1 EXPRESSION; IDIOTYPE VACCINATION; IMMUNE-RESPONSES; CARCINOMA-CELLS C1 NCI, Dept Expt Transplantat & Immunol, Med Branch, Canc Res Ctr, Bethesda, MD 20892 USA. RP Ruffini, PA (reprint author), NCI, POB B,Bldg 567,Room 207, Frederick, MD 21702 USA. NR 72 TC 12 Z9 13 U1 0 U2 0 PU W B SAUNDERS CO PI PHILADELPHIA PA INDEPENDENCE SQUARE WEST CURTIS CENTER, STE 300, PHILADELPHIA, PA 19106-3399 USA SN 0037-1963 J9 SEMIN HEMATOL JI Semin. Hematol. PD JUL PY 2001 VL 38 IS 3 BP 260 EP 267 DI 10.1053/shem.2001.26004 PG 8 WC Hematology SC Hematology GA 458TG UT WOS:000170210200009 PM 11486314 ER PT J AU Russell, SJ Dunbar, CE AF Russell, SJ Dunbar, CE TI Gene therapy approaches for multiple myeloma SO SEMINARS IN HEMATOLOGY LA English DT Article ID BONE-MARROW CELLS; HEMATOPOIETIC STEM-CELLS; IN-VIVO; RETROVIRAL VECTORS; TRANSPLANTED MICE; TUMOR-GROWTH; IDIOTYPIC VACCINATION; MEDIATED TRANSFER; HUMAN MDR1; ADENOVIRUS C1 Mayo Clin & Mayo Fdn, Program Mol Med, Rochester, MN 55905 USA. NHLBI, Hematol Branch, Mol Hematopoiesis Sect, Bethesda, MD 20892 USA. RP Russell, SJ (reprint author), Mayo Clin & Mayo Fdn, Program Mol Med, 200 1st St SW, Rochester, MN 55905 USA. NR 61 TC 4 Z9 4 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 0037-1963 J9 SEMIN HEMATOL JI Semin. Hematol. PD JUL PY 2001 VL 38 IS 3 BP 268 EP 275 DI 10.1016/S0037-1963(01)90019-8 PG 8 WC Hematology SC Hematology GA 458TG UT WOS:000170210200010 PM 11486315 ER PT J AU Yudt, MR Koide, S AF Yudt, MR Koide, S TI Preventing estrogen receptor action with dimer-interface peptides SO STEROIDS LA English DT Article DE estrogen receptor; steroid; helical peptides; protein dimerization; protein aggregation; rational design ID LIGAND-BINDING DOMAIN; DNA-BINDING; TRANSCRIPTIONAL ACTIVATION; SYNTHETIC PEPTIDES; HORMONE RECEPTORS; ANDROGEN RECEPTOR; CRYSTAL-STRUCTURE; THYROID-HORMONE; DIMERIZATION; TAMOXIFEN AB The human estrogen receptor-alpha (hER) is a ligand-activated transcription factor that functions as a homodimer. We sought to further understand the molecular processes involved in dimerization, and to develop a reagent that may function as an antiestrogen independent of the ligand binding site. To this end, we designed a 16-residue 'dimer-interface' oligopeptide derived from the helical region of the hER which is directly involved in dimerization. This peptide, termed the I-box peptide, has: a high helical propensity in aqueous solution. The I-box peptide blocks hER action by causing aggregation and precipitation of both the ligand-bound and apo-hER. This effect is dependent on the helical nature of the peptide. A single Ile to Pro mutation in the helical region of the I-box peptide significantly reduces the helical content and abolishes the precipitation activity. Furthermore, the peptide activity appears to be specific for the hER. The I-box peptide does not significantly affect other proteins or steroid receptors tested. A homologous peptide derived from the nuclear receptor RXR alpha dimer interface, and a LXXLL-containing peptide from the coactivator TIF2 have no detectable in vitro effect on hER function or solubility. Our data suggest that rationally designed molecules capable of affecting steroid receptor quaternary structures may be potential avenues for the development of specific inhibitors of this class of proteins. (C) 2001 Elsevier Science Inc. All rights reserved. C1 Univ Rochester, Sch Med & Dent, Dept Biochem & Biophys, Rochester, NY 14642 USA. RP Koide, S (reprint author), NIEHS, POB 12233, Res Triangle Pk, NC 27709 USA. NR 46 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 0039-128X J9 STEROIDS JI Steroids PD JUL PY 2001 VL 66 IS 7 BP 549 EP 558 DI 10.1016/S0039-128X(00)00224-5 PG 10 WC Biochemistry & Molecular Biology; Endocrinology & Metabolism SC Biochemistry & Molecular Biology; Endocrinology & Metabolism GA 445RD UT WOS:000169470200002 PM 11322963 ER PT J AU Bazzano, LA He, J Ogden, LG Loria, C Vupputuri, S Myers, L Whelton, PK AF Bazzano, LA He, J Ogden, LG Loria, C Vupputuri, S Myers, L Whelton, PK TI Dietary potassium intake and risk of stroke in US men and women - National Health and Nutrition Examination Survey I Epidemiologic Follow-Up Study SO STROKE LA English DT Article DE cardiovascular diseases; cerebrovascular disorders; potassium, dietary; prospective study ID CORONARY HEART-DISEASE; BLOOD-PRESSURE; AFRICAN-AMERICANS; VITAMIN-C; MORTALITY; INFORMATION; MAGNESIUM; DEPLETION; CALCIUM; SMOKING AB Background and Purpose-The few prospective studies that have explored the association between dietary intake of potassium and risk of stroke have reported inconsistent findings. This study examines the relationship between dietary potassium intake and the risk of stroke in a representative sample of the US general population. Methods-Study participants included 9805 US men and women who participated in the first National Health and Nutrition Examination Survey (NHANES I) Epidemiologic Follow-Up Study. Dietary potassium and total energy intake were estimated at baseline by using a 24-hour dietary recall. Incidence data for stroke and coronary heart disease were obtained from medical records and death certificates. Results-Over an average of 19 years of follow up, 927 stroke events and 1847 coronary heart disease events were documented. Overall, stroke hazard was significantly different among quartiles of potassium intake (likelihood ratio P=0.03); however, a test of linear trend across quartiles did not reach a customary level of statistical significance (P=0.14), Participants consuming a low potassium diet at baseline (< 34.6 mmol potassium per day) experienced a 28% higher hazard of stroke (hazard ratio 1.28, 95% CI 1.11 to 1.47; P <0.001) than other participants, after adjustment for established cardiovascular disease risk factors. Conclusions-These findings suggest that low dietary potassium intake is associated with an increased risk of stroke. However, the possibility that the association is due to residual confounding cannot be entirely ruled out in this observational study. C1 Tulane Univ, Sch Publ Hlth & Trop Med, Dept Epidemiol, New Orleans, LA 70112 USA. Tulane Univ, Sch Publ Hlth & Trop Med, Dept Biostat, New Orleans, LA 70112 USA. Tulane Univ, Sch Med, Dept Med, New Orleans, LA 70112 USA. NHLBI, NIH, Bethesda, MD 20892 USA. RP He, J (reprint author), Tulane Univ, Sch Publ Hlth & Trop Med, Dept Epidemiol, 1430 Tulane Ave SL18, New Orleans, LA 70112 USA. FU NHLBI NIH HHS [R03 HL-61954] NR 37 TC 62 Z9 63 U1 0 U2 4 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0039-2499 J9 STROKE JI Stroke PD JUL PY 2001 VL 32 IS 7 BP 1473 EP 1479 PG 7 WC Clinical Neurology; Peripheral Vascular Disease SC Neurosciences & Neurology; Cardiovascular System & Cardiology GA 450CX UT WOS:000169724900001 PM 11441188 ER PT J AU Altarescu, G Moore, DF Pursley, R Campia, U Goldstein, S Bryant, M Panza, JA Schiffmann, R AF Altarescu, G Moore, DF Pursley, R Campia, U Goldstein, S Bryant, M Panza, JA Schiffmann, R TI Enhanced endothelium-dependent vasodilation in Fabry disease SO STROKE LA English DT Article DE acetylcholine; blood flow; endothelium-derived relaxing factors; lipids; nitric oxide; vasodilation ID NITRIC-OXIDE; ARTERIOLAR DILATION; RELAXING FACTOR; L-ARGININE; RELAXATION; MUSCLE; ACETYLCHOLINE; CELLS; MICE; EDHF AB Background and Purpose-Fabry disease is an X-linked lysosomal storage disease secondary to deficiency of alpha -galactosidase A with resulting glycolipid accumulation, particularly globotriaosylceramide in arterial smooth muscle and endothelial cells. A systemic vasculopathy, including early-onset stroke, is prevalent without a clear pathogenesis. Methods-Seventeen normotensive and normocholesterolemic hemizygous Fabry patients (aged 21 to 49 years) and 13 control subjects (aged 21 to 48 years) were investigated by venous plethysmography, allowing assessment of forearm blood flow, Plethysmographic measurements were obtained at baseline and during intra-arterial infusion of acetylcholine and sodium nitroprusside both with and without NG-monomethyl L-arginine (L-NMMA). Results-Forearm blood flow was significantly higher in patients than in control subjects at all 3 acetylcholine doses (P=0.014). Patients had a greater response to acetylcholine even after the addition of L-NMMA (P=0.036). Conclusions-These results demonstrate an increased endothelium-mediated vascular reactivity in Fabry disease. The increased vessel response to acetylcholine with and without L-NMMA suggests altered functionality of non-NO endothelium-dependent vasodilatory pathways. C1 NINDS, Dev & Metab Neurol Branch, NIH, Bethesda, MD 20892 USA. NHLBI, Cardiol Branch, NIH, Bethesda, MD USA. NIH, Ctr Informat Technol, Bethesda, MD 20892 USA. NIH, Off Res Serv, Bethesda, MD 20892 USA. RP Schiffmann, R (reprint author), NINDS, Dev & Metab Neurol Branch, NIH, Bldg 10,Room 3D03,9000 Rockville Pike, Bethesda, MD 20892 USA. FU Intramural NIH HHS [Z99 CT999999] NR 20 TC 94 Z9 98 U1 0 U2 0 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0039-2499 J9 STROKE JI Stroke PD JUL PY 2001 VL 32 IS 7 BP 1559 EP 1562 PG 4 WC Clinical Neurology; Peripheral Vascular Disease SC Neurosciences & Neurology; Cardiovascular System & Cardiology GA 450CX UT WOS:000169724900016 PM 11441201 ER PT J AU Frisch, M Johansen, C Mellemkjaer, L Engels, EA Gridley, G Biggar, RJ Olsen, JH AF Frisch, M Johansen, C Mellemkjaer, L Engels, EA Gridley, G Biggar, RJ Olsen, JH TI Appendectomy and subsequent risk of inflammatory bowel diseases SO SURGERY LA English DT Article ID ULCERATIVE-COLITIS; CROHNS-DISEASE; ACUTE APPENDICITIS; SMOKING; TONSILLECTOMY; DENMARK; CANCER; EPIDEMIOLOGY; REGISTRY; PROTECTS AB Background. Case-control studies have reported an inverse relationship between appendectomy and the risk of ulcerative colitis, but the association has not been confirmed in prospective studies. Methods. Using national hospital discharge registry data in Denmark, the authors followed up 154,434 patients who underwent appendectomy during the period 1977 to 1989 to investigate whether they had subsequent hospitalizations for ulcerative colitis and Crohn's disease. Ratios of observed-to-expected first hospitalizations for inflammatory bowel diseases served as measures of the relative risk (RR). Results. Hospitalization for ulcerative colitis occurred in 84 patients who had appendectomies versus 97.0 expected (RR = 0.87; 95% CI, 0.69-1.09). RRs were not significantly reduced in subgroups defined by sex, age, time since appendectomy, calendar period, or cause of appendectomy. Hospitalization for Crohn's disease occurred in excess (RR = 2.88; 95% CI, 2.45-3.39: n = 150), notably in the first year after appendectomy (RR = 10.83; 95% CI, 8.49-13.62; n = 73); but after 5 year, the RR was not significantly elevated. Conclusions. This large population-based cohort study failed to support a significant inverse association between appendectomy and ulcerative colitis risk in the first decade after the operation. The excess of Crohn's disease shortly after appendectomy most likely reflects differential diagnostic problems in patients newly presenting with abandoned pain. C1 Statens Serum Inst, Danish Epidemiol Sci Ctr, Dept Epidemiol Res, DK-2300 Copenhagen, Denmark. NCI, Div Canc Epidemiol & Genet, Bethesda, MD USA. Inst Canc Epidemiol, Danish Canc Soc, Copenhagen, Denmark. RP Frisch, M (reprint author), Statens Serum Inst, Danish Epidemiol Sci Ctr, Dept Epidemiol Res, 5 Artillerivej, DK-2300 Copenhagen, Denmark. RI Frisch, Morten/E-9206-2016; OI Frisch, Morten/0000-0002-3864-8860; Olsen, Jorgen Helge/0000-0001-9633-5662 FU NCI NIH HHS [N01-CP-85639-04] NR 46 TC 51 Z9 51 U1 0 U2 0 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0039-6060 J9 SURGERY JI Surgery PD JUL PY 2001 VL 130 IS 1 BP 36 EP 43 DI 10.1067/msy.2001.115362 PG 8 WC Surgery SC Surgery GA 453VW UT WOS:000169938000006 PM 11436010 ER PT J AU Cadet, JL Jayanthi, S McCoy, MT Vawter, M Ladenheim, B AF Cadet, JL Jayanthi, S McCoy, MT Vawter, M Ladenheim, B TI Temporal profiling of methamphetamine-induced changes in gene expression in the mouse brain: Evidence from cDNA array SO SYNAPSE LA English DT Article DE cDNA array; neurotoxicity; apoptosis; transcription factors ID DISMUTASE TRANSGENIC MICE; INDUCED NEUROTOXICITY; INDUCED APOPTOSIS; KAPPA-B; DOPAMINE; PROTEIN; DISEASE; SURVIVAL; FAMILY; STRESS AB Methamphetamine (METH) is a neurodegenerative drug of abuse. Its toxicity is characterized by destruction of monoaminergic terminals and by apoptosis in cortical and striatal cell bodies. Multiple factors appear to control METH neurotoxicity, including free radicals and transcription factors. Here, using cDNA arrays, we show the temporal profile of gene expression patterns in the cortex of mice treated with this drug. We obtained two patterns of changes from 588 genes surveyed. First, an early pattern is characterized by upregulation of transcription factors, including members of the jun family. Second, a delayed pattern includes genes related to cell death and to DNA repair. A number of trophic factors were also activated at the later timepoint. These observations suggest that METH can activate a multigene machinery that participates in the production of its toxic effects. The resulting degenerative effects of the drug are thus the result of a balance between protoxic and antiapoptotic mechanisms triggered by its administration to these animals. These observations are of clinical relevance because of the recent identification of degenerative changes in the brains of METH abusers. Synapse 41:40-48, 2001. (C) 2001 Wiley-Liss, Inc. C1 NIDA, Mol Neuropsychiat Sect, NIH, Intramural Res Program, Baltimore, MD 21224 USA. RP Cadet, JL (reprint author), NIDA, Mol Neuropsychiat Sect, NIH, Intramural Res Program, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 58 TC 70 Z9 72 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 0887-4476 J9 SYNAPSE JI Synapse PD JUL PY 2001 VL 41 IS 1 BP 40 EP 48 DI 10.1002/syn.1058 PG 9 WC Neurosciences SC Neurosciences & Neurology GA 436FT UT WOS:000168926400005 PM 11354012 ER PT J AU Huizing, M Anikster, Y Gahl, WA AF Huizing, M Anikster, Y Gahl, WA TI Hermansky-Pudlak syndrome and Chediak-Higashi syndrome: Disorders of vesicle formation and trafficking SO THROMBOSIS AND HAEMOSTASIS LA English DT Article; Proceedings Paper CT XVIIIth Congress of the International-Society-on-Thrombosis-and-Haemostasis CY JUL 06-12, 2001 CL PARIS, FRANCE SP Int Soc Thrombosis & Haemostasis DE albinism; platelet dense bodies; giant lysosomes; LYST; intracellular trafficking ID AP-3 ADAPTER COMPLEX; PIGMENT GRANULE BIOGENESIS; BONE-MARROW-TRANSPLANTATION; STORAGE POOL DEFICIENCY; RAPID DOWN-REGULATION; VON-WILLEBRAND-FACTOR; SYNDROME BEIGE MOUSE; PROTEIN KINASE-C; PALE EAR EP; NATURAL-KILLER AB The rare autosomal recessive metabolic disorders Hermanky-Pudlak syndrome (HPS) and Chediak-Higashi syndrome (CHS) share the clinical findings of oculocutaneous albinism and a platelet storage pool deficiency. In addition, HPS exhibits ceroid lipofuscinosis and CHS is characterized by infections and an accelerated phase. The two disorders result from defects in vesicles of lysosomal lineage. Of the two known HPS-causing genes, HPS1 has no recognizable function, while ADTB3A codes for a subunit of an adaptor complex responsible for new vesicle formation from the trans-Golgi network. Other HPS-causing genes are likely to exist. The only known CHS-causing gene, LYST, codes for a large protein of unknown function. In general, HPS appears to be a disorder of vesicle formation and CHS a defect in vesicle trafficking. These diseases and their variants mirror a group of mouse hypopigmentation mutants. The gene products involved will reveal how the melanosome, platelet dense body, and lysosome are formed and trafficked within cells. C1 NICHD, Sect Human Biochem Genet, Heritable Disorders Branch, NIH, Bethesda, MD 20892 USA. RP Gahl, WA (reprint author), NICHD, Sect Human Biochem Genet, Heritable Disorders Branch, NIH, 10 Ctr Dr,MSC 1830,Bldg 10,Room 9S-241, Bethesda, MD 20892 USA. EM bgahl@helix.nih.gov NR 145 TC 69 Z9 72 U1 1 U2 6 PU F K SCHATTAUER VERLAG GMBH PI STUTTGART PA P O BOX 10 45 43, LENZHALDE 3, D-70040 STUTTGART, GERMANY SN 0340-6245 J9 THROMB HAEMOSTASIS JI Thromb. Haemost. PD JUL PY 2001 VL 86 IS 1 BP 233 EP 245 PG 13 WC Hematology; Peripheral Vascular Disease SC Hematology; Cardiovascular System & Cardiology GA 454GV UT WOS:000169965600029 PM 11487012 ER PT J AU Behrens, RJ Levi, AW Westra, WH Dutta, D Cooper, DS AF Behrens, RJ Levi, AW Westra, WH Dutta, D Cooper, DS TI Langerhans cell histiocytosis of the thyroid: A report of two cases and review of the literature SO THYROID LA English DT Review ID NEEDLE ASPIRATION CYTOLOGY; PRIMARY HYPOTHYROIDISM; INVOLVEMENT; GLAND; CARCINOMA; THERAPY; DISEASE; ADULTS AB Langerhans cell histiocytosis (LCH) is a monoclonal disease of histiocytes that can involve many or very few organ systems. It is a relatively benign disorder with a 3% mortality in adults. LCH rarely involves the thyroid gland. We report two cases, both presenting in males with a goiter. Both patients were treated with subtotal thyroidectomy. The first patient also received radiotherapy to his thyroid bed and scalp. We summarize the prior reported cases of LCH involving the thyroid and review the current treatment modalities used for LCH. C1 Sinai Hosp, Div Endocrinol, Baltimore, MD 21215 USA. Johns Hopkins Univ, Sch Med, Baltimore, MD USA. Natl Canc Inst, Bethesda, MD USA. Johns Hopkins Univ, Sch Med, Dept Pathol, Baltimore, MD 21205 USA. Sinai Hosp, Dept Pathol, Baltimore, MD 21215 USA. RP Cooper, DS (reprint author), Sinai Hosp, Div Endocrinol, 2401 W Belvedere Ave, Baltimore, MD 21215 USA. NR 40 TC 21 Z9 27 U1 1 U2 3 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1050-7256 J9 THYROID JI Thyroid PD JUL PY 2001 VL 11 IS 7 BP 697 EP 705 DI 10.1089/105072501750362781 PG 9 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 455MT UT WOS:000170031200013 PM 11484900 ER PT J AU Hayashi, SM Hong, HHL Toyoda, K Ton, TVT Devereux, TR Maronpot, RR Huff, J Sills, RC AF Hayashi, SM Hong, HHL Toyoda, K Ton, TVT Devereux, TR Maronpot, RR Huff, J Sills, RC TI High frequency of Ras mutations in forestomach and lung tumors of B6C3F1 mice exposed to 1-amino-2,4-dibromoanthraquinone for 2 years SO TOXICOLOGIC PATHOLOGY LA English DT Article DE 1-Amino-2,4-dibromoanthraquinone; forestomach tumor; lung tumor; squamous cell papilloma and carcinoma; alveolar/bronchiolar adenoma and carcinoma; H-ras; K-ras; polymerase chain reaction; restriction fragment length polymorphism; single-stranded conformational polymorphism; analysis ID POLYMERASE CHAIN-REACTION; MOUSE; DNA; ACTIVATION; CHEMICALS; NEOPLASMS; PROTOONCOGENE; RESISTANT; TRANSGENE; TISSUE AB 1-Amino-2,4-dibromoanthraquinone (ADBAQ) is an anthraquinone-derived vat dye, and a potent carcinogen in laboratory animals. In a 2-year study with dietary exposure to 10, 000 or 20, 000 ppm ADBAQ, increased incidence of forestomach and lung tumors were observed in B6C3F(1) mice. The present study indentified genetic alterations in H-ras and K-ras proto-oncogenes in ADBAQ-induced tumors. Point mutations in ras proto-oncogenes were identified by restriction fragment length polymorphism, single-stranded conformational polymorphism analysis and cycle sequencing of polymerase chain reaction-amplified DNA isolated from paraffin-embedded squamous cell papillomas and carcinomas in the forestomach, and alveolar/bronchiolar adenomas and carcinomas in the lung. A higher frequency of ras mutations was identified in ADBAQ-induced forestomach (23/32, 72%) and lung tumors (16/23, 70%) than in spontaneous forestomach (4/11, 36%) and lung tumors (26/86, 30%). H-ras codon 61 CTA mutations were detected in (4/8, 50%) ADBAQ-induced forestomach squamous cell papillomas and (10/24, 42%) squamous cell carcinomas, but not in the spontaneous forestomach tumors examined. H-ras codon 61 CGA mutation (6/24, 25%) was also detected in ADBAQ-induced forestomach squamous cell carcinomas. K-ras codon 61 A to T transversions and A to G transitions were prominent in ADBAQ-induced lung alveolar/bronchiolar adenomas and alveolar/bronchiolar carcinomas. The major finding of A to T transversions or A to G transitions in forestomach and lung tumors suggests that ADBAQ or its metabolites target adenine bases in the ras proto-oncogenes and that these mutations play a dominant role in multi-organ carcinogenesis in the B6C3F(1) mouse. C1 NIEHS, Lab Expt Pathol, Res Triangle Pk, NC 27709 USA. NIEHS, Mol Toxicol Lab, Res Triangle Pk, NC 27709 USA. RP Sills, RC (reprint author), NIEHS, Lab Expt Pathol, POB 12233, Res Triangle Pk, NC 27709 USA. NR 35 TC 9 Z9 10 U1 0 U2 0 PU TAYLOR & FRANCIS INC PI PHILADELPHIA PA 325 CHESTNUT ST, SUITE 800, PHILADELPHIA, PA 19106 USA SN 0192-6233 J9 TOXICOL PATHOL JI Toxicol. Pathol. PD JUL PY 2001 VL 29 IS 4 BP 422 EP 429 DI 10.1080/01926230152499908 PG 8 WC Pathology; Toxicology SC Pathology; Toxicology GA 558DF UT WOS:000175950300004 PM 11560247 ER PT J AU Ozaki, K Mahler, JF Haseman, JK Moomaw, CR Nicolette, ML Nyska, A AF Ozaki, K Mahler, JF Haseman, JK Moomaw, CR Nicolette, ML Nyska, A TI Unique renal tubule changes induced in rats and mice by the peroxisome proliferators 2,4-dichlorophenoxyacetic acid (2,4-D) and WY-14643 SO TOXICOLOGIC PATHOLOGY LA English DT Article DE kidney; peroxisome proliferators; rats and mice; tubular hyperplasia ID ACTIVATED RECEPTOR-ALPHA; CHRONIC DIETARY TOXICITY; 2,4,5-TRICHLOROPHENOXYACETIC ACID; TISSUE DISTRIBUTION; BETA-OXIDATION; LIVER; INDUCTION; KIDNEY; EXPRESSION; ENZYMES AB Peroxisome proliferators are non-mutagenic carcinogens in the liver of rodents, acting both as initiators and promoters. The National Toxicology Program (NTP) conducted a study of several peroxisome proliferators (PPs), including Wyeth (WY)-14643 as a prototypical PP and 2,4-dichlorophenoxyaceti c acid (2,4-D) as a weak PP, in Sprague-Dawley rats, B6C3F1 mice, and Syrian hamsters. In the kidney, an unusual change was observed in the outer stripe of the outer medulla, especially in rats treated with 2,4-D or WY-14643. This change was characterized by foci of tubules that were partially or completely lined by basophilic epithelial cells with decreased cytoplasm and high nuclear density. Changes typical of chronic nephropathy such as interstitial fibrosis or basement membrane thickening were not associated with these foci. Results of immunohistochemical staining for catalase and cytochrome P-450 4A in the kidney indicated increased staining intensity in renal tubular epithelial cells primarily in the region where the affected tubules were observed; however, the altered cells were negative for both immunohistochemical markers. Ultrastructurally, affected cells had long brush borders typical of the P3 tubule segment. The most distinguishing ultrastructural change was a decreased amount of electronlucent cytoplasm that contained few differentiated organelles and, in particular, a prominent reduced volume and number of mitochondria; changes in peroxisomes were not apparent. In addition to the lesion in rats, mice treated with the highest dose of 2, 4-D, but not WY-14643, manifested similar renal tubular changes as seen by light microscopy. Neither chemical induced renal tubular lesions in hamsters. Hepatocellular changes characteristic of PPs were present in all 3 species treated with WY-14643, but not 2,4-D. These results indicate that the rat is the species most sensitive to the nephrotoxic effects of PPs and there is a site specificity to this toxicity related to areas of PP-related enzyme induction. Although 2, 4-D is considered a weak PP for the liver, it was the most effective at inducing renal lesions, indicating that the toxic potency of various PPs will depend on the target organ. C1 NIEHS, Res Triangle Pk, NC 27709 USA. RP Nyska, A (reprint author), NIEHS, POB 12233, Res Triangle Pk, NC 27709 USA. NR 38 TC 13 Z9 13 U1 2 U2 2 PU TAYLOR & FRANCIS INC PI PHILADELPHIA PA 325 CHESTNUT ST, SUITE 800, PHILADELPHIA, PA 19106 USA SN 0192-6233 J9 TOXICOL PATHOL JI Toxicol. Pathol. PD JUL PY 2001 VL 29 IS 4 BP 440 EP 450 DI 10.1080/01926230152499791 PG 11 WC Pathology; Toxicology SC Pathology; Toxicology GA 558DF UT WOS:000175950300006 PM 11560249 ER PT J AU Suwa, T Nyska, A Peckham, JC Hailey, JR Mahler, JF Haseman, JK Maronpot, RR AF Suwa, T Nyska, A Peckham, JC Hailey, JR Mahler, JF Haseman, JK Maronpot, RR TI A retrospective analysis of background lesions and tissue accountability for male accessory sex organs in Fischer-344 rats SO TOXICOLOGIC PATHOLOGY LA English DT Article DE accessory male sex glands; background data; histopathology; rodents; pituitary gland adenoma; prolactin; spontaneous ID LATERAL PROSTATE; F344 RAT; INFLAMMATION; GLAND; HYPERPLASIA; METAPLASIA; EXPOSURE AB Because the paired lobes (ventral, dorsal, lateral, and anterior) of the rat prostate have not been consistently sampled in many carcinogenicity and toxicity studies, comparison among different investigations has been compromised. The lack of specific site identification for prostatic lesions further lessens the value of incidences reported. We present here the lobe-specific incidences and degree of severity of background prostatic, seminal vesicular, and ampullary glandular lesions in 1768 control Fischer-344 rats from 35 recent National Toxicology Program 2-year carcinogenicity and toxicity studies conducted in 4 laboratories. The dorsal and lateral lobes were combined and considered the dorsolateral lobe where inflammation, epithelial degeneration, mucinous cysts, and edema were observed. Inflammation in the dorsolateral lobes was significantly associated with pituitary gland adenoma whose prolactin was suggested to play an important role in pathogenesis of prostatic inflammation. Epithelial degeneration, epithelial hyperplasia, inflammation, edema, and adenoma were conspicuous in the ventral lobes. Inflammation and edema occurred in the anterior lobes (coagulating glands). Inflammation, dilatation, epithelial hyperplasia, edema, and adenoma were observed in the seminal vesicles. Inflammation was also present in the ampullary glands. We suggest an optimal embedment and trimming method in rat prostate and seminal vesicle to ensure adequate, consistent sampling. C1 NIEHS, Lab Expt Pathol, Res Triangle Pk, NC 27709 USA. NIEHS, Biostat Branch, Res Triangle Pk, NC 27709 USA. Expt Pathol Labs Inc, Res Triangle Pk, NC 27709 USA. RP Nyska, A (reprint author), NIEHS, Lab Expt Pathol, POB 12233, Res Triangle Pk, NC 27709 USA. NR 49 TC 17 Z9 17 U1 0 U2 0 PU TAYLOR & FRANCIS INC PI PHILADELPHIA PA 325 CHESTNUT ST, SUITE 800, PHILADELPHIA, PA 19106 USA SN 0192-6233 J9 TOXICOL PATHOL JI Toxicol. Pathol. PD JUL PY 2001 VL 29 IS 4 BP 467 EP 478 DI 10.1080/01926230152500086 PG 12 WC Pathology; Toxicology SC Pathology; Toxicology GA 558DF UT WOS:000175950300009 PM 11560252 ER PT J AU Tharappel, JC Cunningham, ML Spear, BT Glauert, HP AF Tharappel, JC Cunningham, ML Spear, BT Glauert, HP TI Differential activation of hepatic NF-kappa B in rats and hamsters by the peroxisome proliferators Wy-14,643, gemfibrozil, and dibutyl phthalate SO TOXICOLOGICAL SCIENCES LA English DT Article DE NF-kappa B activation; rats; hamsters; Wy-14,643; gemfibrozil; dibutyl phthalate ID SPECIES-DIFFERENCES; DNA-SYNTHESIS; AGENT GEMFIBROZIL; SYRIAN-HAMSTER; GUINEA-PIG; VITAMIN-E; LIVER; NAFENOPIN; MICE; CIPROFIBRATE AB Nuclear factor-kappaB (NF-kappaB) is an oxidative stress-activated transcription factor involved in the regulation of cell proliferation and apoptosis. We found previously that the peroxisome proliferator ciprofibrate activates NF-kappaB in the livers of rats and mice. These species are sensitive to the hepatocarcinogenic effects of peroxisome proliferators, whereas other species such as Syrian hamsters are not. In the present study we examined the effects of 3 different peroxisome proliferators on NF-kappaB activation in rats and Syrian hamsters, The peroxisome proliferators Wy-14,643, gemfibrozil, and dibutyl phthalate were administered to animals for 6, 34, or 90 days. NF-kappaB activity was determined using electrophoretic mobility-shift assays and confirmed using supershift assays. Wy-14,643 increased the DNA binding activity of NF-kappaB at all 3 time points in rats and produced the highest activation of the 3 chemicals tested. Gemfibrozil and dibutyl phthalate increased NF-kappaB activation to a Lesser extent in rats and not at all times. There were no differences in hepatic NF-kappaB levels between control hamsters and hamsters treated with any of the peroxisome proliferators. This study demonstrates species-specific differences in hepatic NF-kappaB activation by peroxisome proliferators. C1 Univ Kentucky, Grad Ctr Nutr Sci, Lexington, KY 40506 USA. Univ Kentucky, Grad Ctr Toxicol, Lexington, KY 40506 USA. Natl Inst Environm Hlth Sci, Res Triangle Pk, NC USA. Univ Kentucky, Dept Microbiol & Immunol, Lexington, KY 40536 USA. Univ Kentucky, Dept Pathol & Lab Med, Lexington, KY 40536 USA. RP Glauert, HP (reprint author), Univ Kentucky, Grad Ctr Nutr Sci, Lexington, KY 40506 USA. OI Spear, Brett/0000-0002-4343-9393 FU NCI NIH HHS [CA-01688, CA-74147]; NIEHS NIH HHS [ES-07266] NR 37 TC 24 Z9 25 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 1096-6080 J9 TOXICOL SCI JI Toxicol. Sci. PD JUL PY 2001 VL 62 IS 1 BP 20 EP 27 DI 10.1093/toxsci/62.1.20 PG 8 WC Toxicology SC Toxicology GA 444KQ UT WOS:000169399000005 PM 11399789 ER PT J AU Latendresse, JR Newbold, RR Weis, CC Delclos, KB AF Latendresse, JR Newbold, RR Weis, CC Delclos, KB TI Polycystic kidney disease induced in F-1 Sprague-Dawley rats fed para-nonylphenol in a soy-free, casein-containing diet SO TOXICOLOGICAL SCIENCES LA English DT Article DE polycystic kidney; nonylphenol; soy-free diet; endocrine disruptor ID PHYTOESTROGEN CONTENT; INHIBITION; GENISTEIN; TOXICITY; EXPOSURE; PROTEIN; LEVEL; CELLS AB para-Nonylphenol (NP; CAS #84852-15-3); an alkylphenol with a 9-carbon olefin side chain, is widely used in the manufacture of nonionic surfactants, lubricant additives, polymer stabilizers, and antioxidants, Due to its wide commercial use and putative endocrine activity in humans and wildlife, the NTP elected to assess its effects on reproduction in multigenerational studies. To avoid known estrogenic activity of phytoestrogens in soy and alfalfa, a soy- and alfalfa-free, casein-containing diet was used in a range-finding study to determine the doses of NP to be tested further. NP was administered to Sprague-Dawley rats in the diet at 0, 5, 25, 200, 500, 1000, or 2000 ppm to F-0 dams beginning on gestation-day 7, The F-1 pups were weaned at postnatal day (PND) 21, and their exposure via diet was continued at the same dose level as their respective dams. Pup weights from birth through weaning were not significantly different from controls in any dose group, but the average weight of both sexes was significantly less compared to controls, beginning with the PND 28 weighing. The F-1 rats were sacrificed on PND 50 (n = 15, 3 pups of each sex from 5 litters for all dose groups). Terminal body weights of males and females in the 2000-ppm dose group were 74% and 85% of controls, respectively. Severe polycystic kidney disease (PKD) was present in 100% of the 2000 ppm-exposed male and female rats. At 1000 ppm, 67% of males and 53% of females had mild to moderate PKD versus none of either sex in the control and lower-dose groups. The no-adverse-effect level (NOAEL) for PKD was determined to be 500 ppm. Previous studies with comparable duration and route of exposure, but using soy-containing diets, reported either no or only mild PKD at 2000 ppm NP. We conclude that the renal toxicity of NP is highly dependent on the diet on which the animals are maintained, The potential interaction of diet and test compounds on nonreproductive as well as reproductive endpoints should be considered when contemplating the use of special diets formulated to minimize exogenous "hormone" content for the study of the effects of putative endocrine disruptive chemicals. C1 Pathol Associates Int, Natl Ctr Toxicol Res, Jefferson, AR 72079 USA. Natl Ctr Toxicol Res, Div Biochem Toxicol, Jefferson, AR 72079 USA. NIEHS, Dev Endocrinol Sect, Reprod Toxicol Grp, Lab Toxicol,Environm Toxicol Program, Res Triangle Pk, NC 27709 USA. RP Latendresse, JR (reprint author), Pathol Associates Int, Natl Ctr Toxicol Res, POB 26,3900 NCTR Rd, Jefferson, AR 72079 USA. RI Latendresse, John/A-9215-2009 NR 36 TC 24 Z9 24 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 1096-6080 J9 TOXICOL SCI JI Toxicol. Sci. PD JUL PY 2001 VL 62 IS 1 BP 140 EP 147 DI 10.1093/toxsci/62.1.140 PG 8 WC Toxicology SC Toxicology GA 444KQ UT WOS:000169399000017 PM 11399801 ER PT J AU Bustin, M AF Bustin, M TI Chromatin unfolding and activation by HMGN chromosomal proteins SO TRENDS IN BIOCHEMICAL SCIENCES LA English DT Review ID NUCLEOSOMAL BINDING DOMAIN; MOBILITY GROUP PROTEIN-14; REMODELING COMPLEXES; NASCENT CHROMATIN; HISTONE H3; HMG-17; TRANSCRIPTION; DNA; CELLS; ENHANCEMENT AB The high mobility group N (HMGN) proteins are a family of nuclear proteins that binds to nucleosomes. changes the architecture of chromatin, and enhances transcription and replication from chromatin templates. The intracellular organization of the HMGN (previously known as HMG-14/17) proteins is dynamic and is related to both cell-cycle and transcriptional events. These proteins roam the nucleus, perhaps as part of multiprotein complexes, and their target interactions are modulated by posttranslational modifications. Functional studies on HMGN proteins provide insights into the molecular mechanisms by which structural proteins affect DNA-dependent activities in the context of chromatin. C1 NCI, Prot Sect, Met Lab, Div Basic Sci,NIH, Bethesda, MD 20892 USA. RP Bustin, M (reprint author), NCI, Prot Sect, Met Lab, Div Basic Sci,NIH, Bethesda, MD 20892 USA. RI Bustin, Michael/G-6155-2015 NR 60 TC 120 Z9 130 U1 0 U2 5 PU ELSEVIER SCIENCE LONDON PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0968-0004 J9 TRENDS BIOCHEM SCI JI Trends Biochem.Sci. PD JUL PY 2001 VL 26 IS 7 BP 431 EP 437 DI 10.1016/S0968-0004(01)01855-2 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 453DZ UT WOS:000169902400018 PM 11440855 ER PT J AU Yewdell, JW AF Yewdell, JW TI Not such a dismal science: the economics of protein synthesis, folding, degradation and antigen processing SO TRENDS IN CELL BIOLOGY LA English DT Editorial Material ID IN-VIVO; MOLECULAR CHAPERONES; 26S PROTEASOME; PA28; POLYPEPTIDES; DYNAMICS; CYTOSOL; SYSTEM; SIGNAL; CELLS AB There is a pronounced tendency among cell biologists to focus on qualitative aspects of cell physiology. The remarkable accomplishments of evolution in creating cells can only be fully appreciated, however, by combining this qualitative analysis with a quantitative assessment of cellular constituents and processes. Here, I consider the overall protein economy of cells as it relates to recent advances in understanding protein folding, ubiquitin-targeted proteasome-mediated degradation of proteins and the generation of peptide ligands for major histocompatibility complex (MHC) class I molecules. C1 NIAID, Cellular Biol Sect, Viral Dis Lab, Bethesda, MD 20892 USA. RP Yewdell, JW (reprint author), NIAID, Cellular Biol Sect, Viral Dis Lab, Bethesda, MD 20892 USA. RI yewdell, jyewdell@nih.gov/A-1702-2012 NR 29 TC 105 Z9 109 U1 0 U2 6 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 JUL PY 2001 VL 11 IS 7 BP 294 EP 297 DI 10.1016/S0962-8924(01)02030-X PG 4 WC Cell Biology SC Cell Biology GA 445MJ UT WOS:000169461100020 PM 11413040 ER PT J AU Cizza, G Ravn, P Chrousos, GP Gold, PW AF Cizza, G Ravn, P Chrousos, GP Gold, PW TI Depression: a major, unrecognized risk factor for osteoporosis? SO TRENDS IN ENDOCRINOLOGY AND METABOLISM LA English DT Review ID BONE-MINERAL DENSITY; POSTMENOPAUSAL WOMEN; BIPOLAR DISORDER; LITHIUM-THERAPY; OLDER-PEOPLE; STRESS; DISEASE; FALLS; INTERLEUKIN-6; METAANALYSIS AB Existing studies of the relationship between depression and osteoporosis have been heterogeneous in their design and use of diagnostic instruments for depression, which might have contributed to the different results on the comorbidity of these two conditions. Nevertheless, these studies reveal a strong association between depression and osteoporosis. Endocrine factors such as depression-induced hypersecretion of corticotropin-releasing hormone and hypercortisolism, hypogonadism, growth hormone deficiency and increased concentration of circulating interleukin 6, might play a crucial role in the bone loss observed in subjects suffering from major depression. C1 NIMH, Clin Neuroendocrinol Branch, Bethesda, MD 20892 USA. Ctr Clin & Basic Res, Ballerup, Denmark. NICHD, Pediat & Reprod Endocrinol Branch, Bethesda, MD USA. RP Cizza, G (reprint author), NIMH, Clin Neuroendocrinol Branch, Bethesda, MD 20892 USA. NR 48 TC 98 Z9 100 U1 3 U2 10 PU ELSEVIER SCIENCE LONDON PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 1043-2760 J9 TRENDS ENDOCRIN MET JI Trends Endocrinol. Metab. PD JUL PY 2001 VL 12 IS 5 BP 198 EP 203 DI 10.1016/S1043-2760(01)00407-6 PG 6 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 445ME UT WOS:000169460600004 PM 11397644 ER PT J AU Stojilkovic, SS Koshimizu, TA AF Stojilkovic, SS Koshimizu, TA TI Signaling by extracellular nucleotides in anterior pituitary cells SO TRENDS IN ENDOCRINOLOGY AND METABOLISM LA English DT Review ID PURINERGIC RECEPTOR-CHANNELS; GATED ION CHANNELS; P2X(2) RECEPTOR; PROLACTIN-RELEASE; FUNCTIONAL EXPRESSION; FROG MELANOTROPHS; P2Y(2) RECEPTORS; SENSORY NEURONS; P-2X RECEPTOR; ATP RECEPTORS AB Pituitary cells secrete ATP, which acts as an autocrine and/or paracrine extracellular messenger on two families of purinergic receptors: G-protein-coupled P2Y receptors (P2YRs) and ion-conducting P2X receptors (P2XRs). Lactotrophs and GH(3)-immortalized cells express the P2Y(2)R subtype. Several P2XR subtypes are expressed in pituitary cells. Gonadotrophs and somatotrophs express P2X(2a)R and P2X(2b)R, which occur as heteromeric channels. Lactotrophs and GH, cells express one or more ion-conducting subtypes from among P2X(3)R, P2X(4)R and P2X(7)R in homomeric form. Thyrotrophs and corticotrophs also express P2XRs, but their identification requires further study. Pituitary cells express purinergic pi receptors, which are activated by adenosine. The A(1)R subtype of these receptors is expressed in melanotrophs and GH, cells. In this review, we briefly discuss the expression and coupling of A(1)R and P2Y(2)R, and focus on the expression and Ca(2+) signaling of P2XRs. C1 NICHHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. RP Stojilkovic, SS (reprint author), NICHHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. EM stankos@helix.nih.gov OI Koshimizu, Taka-aki/0000-0001-5292-7535 NR 47 TC 27 Z9 27 U1 0 U2 0 PU ELSEVIER SCIENCE LONDON PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 1043-2760 J9 TRENDS ENDOCRIN MET JI Trends Endocrinol. Metab. PD JUL PY 2001 VL 12 IS 5 BP 218 EP 225 DI 10.1016/S1043-2760(01)00387-3 PG 8 WC Endocrinology & Metabolism SC Endocrinology & Metabolism GA 445ME UT WOS:000169460600007 PM 11397647 ER PT J AU Shabalina, SA Ogurtsov, AY Kondrashov, VA Kondrashov, AS AF Shabalina, SA Ogurtsov, AY Kondrashov, VA Kondrashov, AS TI Selective constraint in intergenic regions of human and mouse genomes SO TRENDS IN GENETICS LA English DT Article ID SEQUENCE-ANALYSIS; C-ELEGANS; DNA; EVOLUTION; SYNTENY; RODENT AB We aligned and analyzed 100 pairs of complete, orthologous intergenic regions from the human and mouse genomes (average length similar to 12 000 nucleotides). The alignments alternate between highly similar segments and dissimilar segments, indicating a wide variation of selective constraint. The average number of selectively constrained nucleotides within a mammalian intergenic region is at least 2000. This is threefold higher than within a nematode intergenic region and at least twofold higher than the number of selectively constrained nucleotides coding for an average protein. Because mammals possess only two- to threefold more proteins than Caenorhabditis elegans, the higher complexity of mammals might be primarily because of the functioning of intergenic DNA. C1 NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA. Simons Rock Coll, Great Barrington, MA 01230 USA. RP Shabalina, SA (reprint author), NIH, Natl Ctr Biotechnol Informat, Natl Lib Med, Bethesda, MD 20894 USA. EM shabalin@ncbi.nlm.nih.gov RI Shabalina, Svetlana/N-8939-2013 OI Shabalina, Svetlana/0000-0003-2272-7473 NR 19 TC 102 Z9 103 U1 0 U2 4 PU ELSEVIER SCIENCE LONDON PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0168-9525 J9 TRENDS GENET JI Trends Genet. PD JUL PY 2001 VL 17 IS 7 BP 373 EP 376 DI 10.1016/S0168-9525(01)02344-7 PG 4 WC Genetics & Heredity SC Genetics & Heredity GA 449GA UT WOS:000169674900004 PM 11418197 ER PT J AU Marinissen, MJ Gutkind, JS AF Marinissen, MJ Gutkind, JS TI G-protein-coupled receptors and signaling networks: emerging paradigms SO TRENDS IN PHARMACOLOGICAL SCIENCES LA English DT Review ID HETEROTRIMERIC G-PROTEINS; EPIDERMAL GROWTH-FACTOR; G-BETA-GAMMA; CONSTITUTIVELY ACTIVATING MUTATION; RAS-DEPENDENT ACTIVATION; JUN NH2-TERMINAL KINASE; SRC TYROSINE KINASE; CELL-PROLIFERATION; MAP KINASE; LYSOPHOSPHATIDIC ACID AB G-protein-coupled receptors (GPCRs) constitute the largest family of cell-surface molecules involved in signal transmission, These receptors play key physiological roles and their dysfunction results in several diseases. Recently, it has been shown that many of the cellular responses mediated by GPCRs do not involve the sole stimulation of conventional second-messenger-generating systems, but instead result from the functional integration of an intricate network of intracellular signaling pathways. Effecters for GPCRs that are independent of G proteins have now also been identified, thus changing the conventional view of the GPCR-heterotrimeric-G-protein-associated effector. The emerging information is expected to help elucidate the most basic mechanism by which these receptors exert their numerous physiological roles, in addition to determining why the perturbation of their function results in many pathological conditions. C1 NIDCR, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD 20892 USA. RP Marinissen, MJ (reprint author), NIDCR, Oral & Pharyngeal Canc Branch, NIH, 30 Convent Dr,Bldg 30,Room 211, Bethesda, MD 20892 USA. EM mmarinissen@dir.nidcr.nih.gov; sg39v@nih.gov RI Gutkind, J. Silvio/A-1053-2009 NR 75 TC 635 Z9 652 U1 12 U2 102 PU ELSEVIER SCIENCE LONDON PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0165-6147 J9 TRENDS PHARMACOL SCI JI Trends Pharmacol. Sci. PD JUL PY 2001 VL 22 IS 7 BP 368 EP 376 DI 10.1016/S0165-6147(00)01678-3 PG 9 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 451QG UT WOS:000169812300011 PM 11431032 ER PT J AU Feleppa, EJ Ennis, RD Schiff, PB Wuu, CS Kalisz, A Ketterling, J Urban, S Liu, T Fair, WR Porter, CR Gillespie, JR AF Feleppa, EJ Ennis, RD Schiff, PB Wuu, CS Kalisz, A Ketterling, J Urban, S Liu, T Fair, WR Porter, CR Gillespie, JR TI Spectrum-analysis and neural networks for imaging to detect and treat prostate cancer SO ULTRASONIC IMAGING LA English DT Article DE biopsy guidance; neural networks; prostate cancer; radiation therapy; spectrum analysis; therapy targeting; ultrasonic imaging ID TISSUE-CHARACTERIZATION; BIOPSY AB Conventional B-mode ultrasound currently is the standard means of imaging the prostate for guiding prostate biopsies and planning brachytherapy to treat prostate cancer. Yet B-mode images do not adequately display cancerous lesions of the prostate. Ultrasonic tissue-type imaging based on spectrum analysis of radiofrequency (rf) echo signals has shown promise for overcoming the limitations of B-mode imaging for visualizing prostate tumors. T his method of tissue-type imaging utilizes nonlinear classifiers, such as neural networks, to classify tissue based on values of spectral parameter and clinical variables. Two- and three-dimensional images based on these methods demonstrate potential for guiding prostate biopsies and targeting radiotherapy of prostate cancer. Two-dimensional images are being generated in real time in ultrasound scanners used for real-time biopsy guidance and have been incorporated into commercial dosimetry software used for brachytherapy planning. Three-dimensional renderings show promise for depicting locations and volumes of cancer foci for disease evaluation to assist staging and treatment planning, and potentially for registration or fusion with CT images for targeting external-beam radiotherapy. C1 Riverside Res Inst, Biomed Engn Labs, New York, NY 10038 USA. Columbia Presbyterian Med Ctr, Dept Radiat Oncol, New York, NY 10032 USA. Haelth, New York, NY 10012 USA. SUNY Stony Brook, Stony Brook Med Ctr, Dept Urol, Stony Brook, NY 11794 USA. NCI, Sci Applicat Int Corp, Bethesda, MD 20892 USA. RP Feleppa, EJ (reprint author), Riverside Res Inst, Biomed Engn Labs, 156 William St, New York, NY 10038 USA. OI Ketterling, Jeffrey/0000-0002-1389-7756 FU NCI NIH HHS [CA53561] NR 11 TC 32 Z9 33 U1 0 U2 2 PU DYNAMEDIA INC PI SILVER SPRING PA 2 FULHAM COURT, SILVER SPRING, MD 20902 USA SN 0161-7346 J9 ULTRASONIC IMAGING JI Ultrason. Imaging PD JUL PY 2001 VL 23 IS 3 BP 135 EP 146 PG 12 WC Acoustics; Engineering, Biomedical; Radiology, Nuclear Medicine & Medical Imaging SC Acoustics; Engineering; Radiology, Nuclear Medicine & Medical Imaging GA 541CU UT WOS:000174968300001 PM 11958585 ER PT J AU Cardiff, RD Wagner, U Henninghausen, L AF Cardiff, RD Wagner, U Henninghausen, L TI Mammary cancer in humans and mice: A tutorial for comparative pathology SO VETERINARY PATHOLOGY LA English DT Editorial Material C1 Univ Calif Davis, Ctr Comparat Med, Davis, CA 95616 USA. NIH, Lab Genet & Physiol, Bethesda, MD 20892 USA. RP Cardiff, RD (reprint author), Univ Calif Davis, Ctr Comparat Med, Davis, CA 95616 USA. RI Wagner, Ulrike/B-6039-2009 NR 1 TC 13 Z9 14 U1 0 U2 0 PU AMER COLL VET PATHOLOGIST PI LAWRENCE PA 810 EAST 10TH STREET, LAWRENCE, KS 66044 USA SN 0300-9858 J9 VET PATHOL JI Vet. Pathol. PD JUL PY 2001 VL 38 IS 4 BP 357 EP 358 DI 10.1354/vp.38-4-357 PG 2 WC Pathology; Veterinary Sciences SC Pathology; Veterinary Sciences GA 450QQ UT WOS:000169754900001 PM 11467469 ER PT J AU Wu, GG Jin, SZ Deng, ZH Zhao, TM AF Wu, GG Jin, SZ Deng, ZH Zhao, TM TI Polymerase chain reaction with sequence-specific primers-based genotyping of the human Dombrock blood group DO1 and DO2 alleles and the DO gene frequencies in Chinese blood donors SO VOX SANGUINIS LA English DT Article ID GROUP SYSTEM; ANTIGENS C1 NIAID, Mol & Cellular Immunogenet Sect, NIH, Bethesda, MD 20892 USA. Shenzhen Inst Transfus Med, Shenzhen, Peoples R China. RP Zhao, TM (reprint author), NIAID, Mol & Cellular Immunogenet Sect, NIH, Bldg 9,Room 1E124,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 8 TC 15 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 0042-9007 J9 VOX SANG JI Vox Sang. PD JUL PY 2001 VL 81 IS 1 BP 49 EP 51 DI 10.1046/j.1423-0410.2001.00052.x PG 3 WC Hematology SC Hematology GA 456NC UT WOS:000170088100010 PM 11520417 ER PT J AU DuVal, G Sartorius, L Clarridge, B Gensler, G Danis, M AF DuVal, G Sartorius, L Clarridge, B Gensler, G Danis, M TI What triggers requests for ethics consultations? SO WESTERN JOURNAL OF MEDICINE LA English DT Article ID GOALS AB Objective To investigate what triggers clinicians' requests for ethics consultations. Design Cross-sectional telephone survey. Setting and participants Randomly selected physicians throughout the United States who practice in internal medicine, oncology, and critical care. Main measurements Sociodemographic characteristics, training in medicine and ethics, and practice characteristics; types of ethical problems that prompt requests for consultation, and factors triggering consultation requests. Results Of 344 responding physicians, 190 (55.2%) reported requesting ethics consultations. Most commonly these were for ethical dilemmas related to end-of-life decision making, patient autonomy issues, and conflict. The most common triggers that led to consultation requests were wanting help resolving a conflict; wanting assistance with interactions with a difficult family, patient, or surrogate; wanting help with making a decision or planning care; and emotional triggers. Physicians who were ethnically in the minority, practiced in communities under 500,000 population, or who were trained in the United States were more likely to request consultations to resolve conflict Conclusions Conflicts and other emotionally charged concerns more commonly trigger consultation requests than other cognitively based concerns. When consulting ethicists need to be prepared to mediate conflicts and handle sometimes difficult emotional situations. The data suggest that ethics consultants might serve clinicians well by consulting on a more proactive basis ro avoid conflicts and by educating clinicians to develop mediation skills. C1 Univ Toronto, Ctr Addict & Mental Hlth, Toronto, ON M5G 1L4, Canada. Univ Toronto, Joint Ctr Bioeth, Toronto, ON M5G 1L4, Canada. NIH, Dept Clin Bioeth, Ctr Clin, Sect Eth & Hlth Policy, Bethesda, MD 20892 USA. Univ Massachusetts, Survey Res Ctr, Boston, MA 02125 USA. EMMES Corp, Potomac, MD USA. RP DuVal, G (reprint author), Univ Toronto, Ctr Addict & Mental Hlth, 88 Coll St, Toronto, ON M5G 1L4, Canada. NR 13 TC 2 Z9 2 U1 0 U2 1 PU B M J PUBLISHING INC PI SAN FRANCISCO PA 221 MAIN ST, PO BOX 7690, SAN FRANCISCO, CA 94120-7690 USA SN 0093-0415 J9 WESTERN J MED JI West. J. Med. PD JUL PY 2001 VL 175 IS 1 BP 24 EP 30 DI 10.1136/ewjm.175.1.24 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 448NG UT WOS:000169632500015 ER PT J AU Curt, GA AF Curt, GA TI Fatigue in cancer - Like pain, this is a symptom that physicians can and should manage SO BRITISH MEDICAL JOURNAL LA English DT Editorial Material C1 NCI, NIH, Ctr Clin, Bethesda, MD 20892 USA. RP Curt, GA (reprint author), NCI, NIH, Ctr Clin, Bethesda, MD 20892 USA. NR 8 TC 35 Z9 35 U1 0 U2 2 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 0959-535X J9 BRIT MED J JI Br. Med. J. PD JUN 30 PY 2001 VL 322 IS 7302 BP 1560 EP 1560 DI 10.1136/bmj.322.7302.1560 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 449QT UT WOS:000169699200005 PM 11431284 ER PT J AU Baird, AE Dambrosia, J Janket, SJ Eichbaum, Q Chaves, C Silver, B Barber, PA Parsons, M Darby, D Davis, S Caplan, LR Edelman, RE Warach, S AF Baird, AE Dambrosia, J Janket, SJ Eichbaum, Q Chaves, C Silver, B Barber, PA Parsons, M Darby, D Davis, S Caplan, LR Edelman, RE Warach, S TI A three-item scale for the early prediction of stroke recovery SO LANCET LA English DT Article ID ISCHEMIC STROKE; CEREBRAL INFARCTION; SCORE; PROGNOSIS; TRIAL AB Background Accurate assessment of prognosis in the first hours of stroke is desirable for best patient management. We aimed to assess whether the extent of ischaemic brain injury on magnetic resonance diffusion-weighted imaging (MR DWI) could provide additional prognostic information to clinical factors. Methods In a three-phase study we studied 66 patients from a North American teaching hospital who had: MR DWI within 36 hours of stroke onset; the National Institutes of Health Stroke Scale (NIHSS) score measured at the time of scanning; and the Barthel Index measured no later than 3 months after stroke. We used logistic regression to derive a predictive model for good recovery. This logistic regression model was applied to an independent series of 63 patients from an Australian teaching hospital, and we then developed a three-item scale for the early prediction of stroke recovery. Findings Combined measurements of the NIHSS score (p=0.01), time in hours from stroke onset to MR DWI (p=0.02), and the volume of ischaemic brain tissue on MR DWI (p=0.04) gave the best prediction of stroke recovery. The model was externally validated on the Australian sample with 0.77 sensitivity and 0.88 specificity. Three likelihood levels for stroke recovery-low (0-2), medium (3-4), and high (5-7)-were identified on the three-item scale. Interpretation The combination of clinical and MR DWI factors provided better prediction of stroke recovery than any factor alone, shortly after admission to hospital. This information was incorporated into a three-item scale for clinical use. C1 NINDS, Bethesda, MD 20892 USA. Vet Affairs Med Ctr, Bedford, MA USA. Harvard Univ, Sch Med, Boston, MA USA. Lahey Clin, Lexington, MA USA. London Hlth Sci Ctr, London, ON, Canada. Auckland Hosp, Auckland, New Zealand. Royal Melbourne Hosp, Melbourne, Vic, Australia. Beth Israel Deaconess Med Ctr, Boston, MA 02215 USA. Evanston NW Healthcare, Evanston, IL USA. RP Baird, AE (reprint author), NINDS, Bethesda, MD 20892 USA. RI Davis, Stephen /B-7344-2009; Davis, Stephen/L-5260-2013; Parsons, Mark/G-3750-2014; OI Davis, Stephen/0000-0003-0962-2300; Barber, P Alan/0000-0003-2469-9023 NR 25 TC 128 Z9 132 U1 1 U2 6 PU LANCET LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0140-6736 J9 LANCET JI Lancet PD JUN 30 PY 2001 VL 357 IS 9274 BP 2095 EP 2099 DI 10.1016/S0140-6736(00)05183-7 PG 5 WC Medicine, General & Internal SC General & Internal Medicine GA 447RQ UT WOS:000169586600014 PM 11445104 ER PT J AU Hager, G AF Hager, G TI Alan Wolffe (1954-2001) - Obituary SO CELL LA English DT Biographical-Item C1 NCI, Bethesda, MD 20892 USA. RP Hager, G (reprint author), NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU CELL PRESS PI CAMBRIDGE PA 1100 MASSACHUSETTES AVE,, CAMBRIDGE, MA 02138 USA SN 0092-8674 J9 CELL JI Cell PD JUN 29 PY 2001 VL 105 IS 7 BP 849 EP 850 DI 10.1016/S0092-8674(01)00413-5 PG 2 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 449BK UT WOS:000169664300005 PM 11439181 ER PT J AU Krakauer, T Li, BQ Young, HA AF Krakauer, T Li, BQ Young, HA TI The flavonoid baicalin inhibits superantigen-induced inflammatory cytokines and chemokines SO FEBS LETTERS LA English DT Article DE immunosuppression; superantigen; cytokine and chemokine; baicalin; flavonoid ID TOXIC-SHOCK-SYNDROME; STAPHYLOCOCCAL-ENTEROTOXIN-A; TUMOR-NECROSIS-FACTOR; HUMAN T-CELLS; ANTIINFLAMMATORY ACTIVITY; HUMAN INTERLEUKIN-1; POTENTIAL ROLE; HUMAN-DISEASE; BINDING; COMPONENTS AB Excessive release of proinflammatory cytokines mediates the rode effect of superantigenic staphylococcal exotoxins (SE). Baicalin, a flavone isolated from the Chinese herb Scutellaria baicalensis Georgi and used in China to treat infectious diseases, inhibited SE-stimulated T-cell proliferation (by 98%) and production of interleukin 1 beta, interleukin 6, tumor necrosis factor, interferon gamma, monocyte chemotactic protein 1, macrophage inflammatory protein (MIP)-1 alpha, and MIP-1 beta mRNA and protein by human peripheral blood mononuclear cells, These data suggest that baicalin may be therapeutically useful for mitigating the pathogenic effects of SE by inhibiting the signaling pathways activated by superantigens, (C) 2001 Federation of European Biochemieal Societies, Published by Elsevier Science B.V. All rights reserved. C1 USA, Med Res Inst Infect Dis, Dept Immunol & Mol Biol, Frederick, MD 21702 USA. SAIC Frederick, Intramural Res Support Program, Frederick, MD 21702 USA. NCI, Expt Immunol Lab, Frederick, MD 21702 USA. RP Krakauer, T (reprint author), USA, Med Res Inst Infect Dis, Dept Immunol & Mol Biol, 1425 Porter St, Frederick, MD 21702 USA. RI Young, Howard/A-6350-2008 OI Young, Howard/0000-0002-3118-5111 NR 29 TC 79 Z9 105 U1 2 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 JUN 29 PY 2001 VL 500 IS 1-2 BP 52 EP 55 DI 10.1016/S0014-5793(01)02584-4 PG 4 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 450BV UT WOS:000169722400010 PM 11434925 ER PT J AU Craigie, R AF Craigie, R TI HIV integrase, a brief overview from chemistry to therapeutics SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Review ID IMMUNODEFICIENCY-VIRUS TYPE-1; TARGET SITE SELECTION; PHOTO-CROSS-LINKING; DNA STRAND TRANSFER; CATALYTIC DOMAIN; CRYSTAL-STRUCTURE; RETROVIRAL INTEGRASES; TERMINAL DOMAIN; ACTIVE-SITE; CORE DOMAIN C1 NIDDK, NIH, Mol Biol Lab, Bethesda, MD 20892 USA. RP Craigie, R (reprint author), NIDDK, NIH, Mol Biol Lab, Bldg 5,Rm 301,5 Ctr Dr,MSC 0560, Bethesda, MD 20892 USA. NR 48 TC 190 Z9 200 U1 0 U2 10 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 JUN 29 PY 2001 VL 276 IS 26 BP 23213 EP 23216 DI 10.1074/jbc.R100027200 PG 4 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 446TW UT WOS:000169531100001 PM 11346660 ER PT J AU Stephen, AG Raval-Fernandes, S Huynh, T Torres, M Kickhoefer, VA Rome, LH AF Stephen, AG Raval-Fernandes, S Huynh, T Torres, M Kickhoefer, VA Rome, LH TI Assembly of vault-like particles in insect cells expressing only the major vault protein SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID VIRUS-LIKE PARTICLES; RIBONUCLEOPROTEIN-PARTICLES; SMALL RNA; RESISTANCE; POLYMERASE; DISRUPTION; COMPLEX; CONTAIN AB Vaults are the largest (13 megadalton) cytoplasmic ribonucleoprotein particles known to exist in eukaryotic cells. They have a unique barrel-shaped structure with 8-fold symmetry. Although the precise function of vaults is unknown, their wide distribution and highly conserved morphology in eukaryotes suggests that their function is essential and that their structure must be important for their function. The 100-kDa major vault protein (MVP) constitutes similar to 75% of the particle mass and is predicted to form the central barrel portion of the vault. To gain insight into the mechanisms for vault assembly, we have expressed rat MVP in the Sf9 insect cell line using a baculovirus vector. Our results show that the expression of the rat MVP alone can direct the formation of particles that have biochemical characteristics similar to endogenous rat vaults and display the distinct vault-like morphology when negatively stained and examined by electron microscopy. These particles are the first example of a single protein polymerizing into a non-spherically, non-cylindrically symmetrical structure. Understanding vault assembly will enable us to design agents that disrupt vault formation and hence aid in elucidating vault function in vivo. C1 Univ Calif Los Angeles, Sch Med, Dept Biol Chem, Los Angeles, CA 90095 USA. Univ Calif Los Angeles, Sch Med, Jonsson Comprehens Canc Ctr, Los Angeles, CA 90095 USA. NCI, Frederick Canc Res & Dev Ctr, SAIC Frederick, Frederick, MD 21702 USA. RP Rome, LH (reprint author), Univ Calif Los Angeles, Sch Med, Dept Biol Chem, 33-131 CHS,10833 Le Conte Ave, Los Angeles, CA 90095 USA. RI Rome, Leonard/E-8786-2016; OI Rome, Leonard/0000-0002-1236-2063; Kickhoefer, Valerie/0000-0002-0048-0580 FU NIGMS NIH HHS [GM38097] NR 23 TC 81 Z9 81 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 JUN 29 PY 2001 VL 276 IS 26 BP 23217 EP 23220 DI 10.1074/jbc.C100226200 PG 4 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 446TW UT WOS:000169531100002 PM 11349122 ER PT J AU Suzuki, N Khoo, KH Chen, HC Johnson, JR Lee, YC AF Suzuki, N Khoo, KH Chen, HC Johnson, JR Lee, YC TI Isolation and characterization of major glycoproteins of pigeon egg white - Ubiquitous presence of unique N-glycans containing Gal alpha 1-4Gal SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID UROPATHOGENIC ESCHERICHIA-COLI; LINKED SUGAR CHAINS; SIALOSYL GALACTOSYL GLOBOSIDE; PREFERRED BINDING-RECEPTOR; AMINO-ACID-SEQUENCE; STREPTOCOCCUS-SUIS; HEN OVALBUMIN; STRUCTURAL CHARACTERIZATION; LIQUID-CHROMATOGRAPHY; BIOSYNTHETIC-PATHWAY AB Ovotransferrin (POT), two ovalbumins (POA(hi) and POA(lo)), and ovomucoid (POM) were isolated from pigeon egg white (PEW). Unlike their chicken egg white counterparts, PEW glycoproteins contain terminal Gal alpha1-4Gal, as evidenced by GS-I lectin (specific for terminal alpha -Gal), anti-P-1 (Gal alpha1-4Gal beta1- 4GlcNAc beta1-3Gal beta1-4Glc beta1-1Cer) monoclonal antibody, and P fimbriae on uropathogenic Escherichia coli (specific for Gal alpha1-4Gal). Gal alpha1-4Gal on PEW glycoproteins were found in N-glycans releasable by treatment with glycoamidase F. The respective contents of N-glycans in each glycoprotein were 3.5%, POT; 17%, POA(hi); and 31-37%, POM. POA(hi) has four N-glycosylation sites, in contrast to chicken ovalbumin, which has only one. High performance liquid chromatography analysis showed that N-glycans on POA(hi) were highly heterogeneous. Mass spectrometric analysis revealed that the major N-glycans were monosialylated tri-, tetra-, and penta-antennary oligosaccharides containing terminal Gal alpha1-4Gal with or without bisecting N-acetylglucosamine, Oligo saccharide chains terminating in Gal alpha1-4Gal are rare among N-glycans from the mammals and avians that have been studied, and our finding is the first predominant presence of (Gal alpha1-4Gal)-terminated N-glycans. C1 Johns Hopkins Univ, Dept Biol, Baltimore, MD 21218 USA. Acad Sinica, Inst Biol Chem, Taipei 115, Taiwan. NICHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. Univ Minnesota, Dept Med, Minneapolis, MN 55417 USA. Vet Affairs Med Ctr, Dept Med, Minneapolis, MN 55417 USA. RP Lee, YC (reprint author), Johns Hopkins Univ, Dept Biol, Baltimore, MD 21218 USA. FU NIDDK NIH HHS [DK-47504, DKO9970] NR 65 TC 34 Z9 36 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 JUN 29 PY 2001 VL 276 IS 26 BP 23221 EP 23229 DI 10.1074/jbc.M101379200 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 446TW UT WOS:000169531100003 PM 11287422 ER PT J AU Xiao, WH Wang, LH Yang, XY Chen, TS Hodge, D Johnson, PF Farrar, W AF Xiao, WH Wang, LH Yang, XY Chen, TS Hodge, D Johnson, PF Farrar, W TI CCAAT/enhancer-binding protein beta mediates interferon-gamma-induced p48 (ISGF3-gamma) gene transcription in human monocytic cells SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID TARGETED DISRUPTION; ISGF3 COMPONENTS; MELANOMA-CELLS; DNA-BINDING; IFN-BETA; ACTIVATION; EXPRESSION; INDUCTION; RESISTANT; FAMILY AB Previous studies have identified a novel interferon-stimulated response element-like element, termed gamma -interferon-activating transcription element, within the interferon-stimulating gene factor-3 gamma (p48) promoter region that is bound by novel transcription factors in response to stimulation with interferons (IFNs) (Weihua, X,, Kolla, V., and Kalvakolanu, D, V. (1997) Proc. Natl. Acad, Sci. U. S. A. 94, 103-108), In the present study, we have identified CCAAT/enhancer-binding protein beta (C/EBP-beta) as one of the gamma -interferon-activating transcription element cognate transcription factors by screening a human monophage-derived cDNA library in a yeast one-hybrid system. Electrophoretic mobility shift assay studies suggest that C/EBP-beta dynamically regulates p48 gene expression upon IFN-gamma stimulation by undergoing changes in its heterodimerization partners. Transient transfection studies demonstrate that overexpression of C/EBP-beta strongly enhanced IFN-gamma -induced transcription from the p48 promoter. However, deletion mutants of C/EBP-beta that lack the N-terminal transactivation domain were unable to stimulate the p48 promoter. Western blotting revealed that C/EBP-beta is induced by IFN-gamma stimulation in THP-1-derived macrophages. Collectively, these results suggest that C/EBP-beta plays an important role in the human IFN-gamma signaling pathway by transcriptional regulation of p48 gene expression, an essential component in the IFN signaling pathway. C1 NCI, Frederick Canc Res & Dev Ctr, Intramural Res Program, SAIC Frederick,NIH, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, Mol Immunoregulat Lab, Cytokine Mol Mech Sect,NIH, Frederick, MD 21702 USA. NCI, Frederick Canc Res & Dev Ctr, Regulat Cell Growth Lab, NIH, Frederick, MD 21702 USA. RP Xiao, WH (reprint author), NCI, Frederick Canc Res & Dev Ctr, Intramural Res Program, SAIC Frederick,NIH, POB B,Bldg 560,Rm 31-76, Frederick, MD 21702 USA. RI Johnson, Peter/A-1940-2012; Chen, Taosheng/I-6351-2013; Xiao, Weihua/N-2775-2013 OI Johnson, Peter/0000-0002-4145-4725; Xiao, Weihua/0000-0001-9102-6326 FU NCI NIH HHS [N0 1-CO-56000] NR 40 TC 17 Z9 17 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 JUN 29 PY 2001 VL 276 IS 26 BP 23275 EP 23281 DI 10.1074/jbc.M010047200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 446TW UT WOS:000169531100010 PM 11312260 ER PT J AU Lim, SE Copeland, WC AF Lim, SE Copeland, WC TI Differential incorporation and removal of antiviral deoxynucleotides by human DNA polymerase gamma SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; ANTI-HIV DEOXYNUCLEOTIDES; MITOCHONDRIAL-DNA; REVERSE-TRANSCRIPTASE; EXONUCLEASE ACTIVITY; NUCLEOSIDE ANALOGS; (-)-2'-DEOXY-3'-THIACYTIDINE 3TC; INTRACELLULAR PHOSPHORYLATION; SUBSTRATE-SPECIFICITY; INSERTION FIDELITY AB Mitochondrial toxicity can result hom antiviral nucleotide analog therapy used to control human immunodeficiency virus type 1 infection. We evaluated the ability of such analogs to inhibit DNA synthesis by the human mitochondrial DNA polymerase (pol gamma) by comparing the insertion and exonucleolytic removal of six antiviral nucleotide analogs. Apparent steady-state K-m and k(cat) values for insertion of 2 ' ,3 ' -dideoxy-TTP (ddTTP), 3 ' -azido-TTP (AZT-TP), 2 ' ,3 ' -dideoxy-TTP (dd-TTP), 2 ' ,3 ' -didehydro-TTP (D4T-TP), (-)-2 ' ,3 ' -dideoxy-3 ' -thiacytidine (3TC-TP), and carbocyclic 2 ' ,3 ' -didehydro-ddGTP (CBV-TP) indicated incorporation of all six analogs, albeit with varying efficiencies. Dideoxynucleotides and D4T-TP were utilized by pol gamma in vitro as efficiently as natural deoxynucleotides, whereas AZT-TP, 3TC-TP, and CBV-TP were only moderate inhibitors of DNA chain elongation. Inefficient excision of dideoxynucleotides, D4T, AZT, and CBV from DNA predicts persistence in vivo following successful incorporation. In contrast, removal of 3'-terminal 3TC residues was 50% as efficient as natural 3 ' termini. Finally, me observed inhibition of exonuclease activity by concentrations of AZT-monophosphate known to occur in cells. Thus, although their greatest inhibitory effects are through incorporation and chain termination, persistence of these analogs in DNA and inhibition of exonucleolytic proofreading may also contribute to mitochondrial toxicity. C1 NIEHS, Mol Genet Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Copeland, WC (reprint author), NIEHS, Mol Genet Lab, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 61 TC 154 Z9 160 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 JUN 29 PY 2001 VL 276 IS 26 BP 23616 EP 23623 DI 10.1074/jbc.M101114200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 446TW UT WOS:000169531100053 PM 11319228 ER PT J AU Tiffany, HL Lavigne, MC Cui, YH Wang, JM Leto, TL Gao, JL Murphy, PM AF Tiffany, HL Lavigne, MC Cui, YH Wang, JM Leto, TL Gao, JL Murphy, PM TI Amyloid-beta induces chemotaxis and oxidant stress by acting at formylpeptide receptor 2, a G protein-coupled receptor expressed in phagocytes and brain SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID FORMYL PEPTIDE RECEPTOR; ALZHEIMERS-DISEASE; PRECURSOR PROTEIN; SCAVENGER RECEPTOR; MICROGLIAL CELLS; HUMAN MONOCYTE; INTERFERON-GAMMA; GENE-CLUSTER; MOUSE; NEUROTOXICITY AB Amyloid-beta, the pathologic protein in Alzheimer's disease, induces chemotaxis and production of reactive oxygen species in phagocytic cells, but mechanisms have not been fully defined. Here me provide three lines of evidence that the phagocyte G protein-coupled receptor (N-formylpeptide receptor 2 (FPR2)) mediates these amyloid-beta -dependent functions in phagocytic cells. First, transfection of FPR2, but not related receptors, including the other known N-formylpeptide receptor FPR, reconstituted amyloid-beta -dependent chemotaxis and calcium flux in HEK 293 cells. Second, amyloid-beta induced both calcium flux and chemotaxis in mouse neutrophils (which express endogenous FPR2) with similar potency as in FPR2-transfected HEK 293 cells, This activity could be specifically desensitized in both cell types by preincubation with a specific FPR2 agonist, which desensitizes the receptor, or with pertussis toxin, which uncouples it from G(i)-dependent signaling. Third, specific and reciprocal desensitization of superoxide production was observed when N-formylpeptides and amyloid-beta were used to sequentially stimulate neutrophils from FPR -/- mice, which express FPR2 normally. Potential biological relevance of these results to the neuroinflamma. tion associated with Alzheimer's disease was suggested by two additional findings: first, FPR2 mRNA could be detected by PCR in mouse brain; second, induction of FPR2 expression correlated with induction of calcium flux and chemotaxis by amyloid-beta in the mouse microglial cell line N9, Further, in sequential stimulation experiments with N9 cells, N-formylpeptides and amyloid-beta were able to reciprocally cross-desensitize each other. Amyloid-beta was also a specific agonist at the human counterpart of FPR2, the FPR-like 1 receptor. These results suggest a unified signaling mechanism for linking amyloid-beta to phagocyte chemotaxis and oxidant stress in the brain. C1 NIAID, Mol Signaling Sect, NIH, Bethesda, MD 20892 USA. NIAID, Genet Immunotherapy Sect, Host Def Lab, NIH, Bethesda, MD 20892 USA. NCI, Frederick Canc Res & Dev Ctr, Mol Immunoregulat Lab, Frederick, MD 21702 USA. RP Murphy, PM (reprint author), NIAID, Mol Signaling Sect, NIH, Bldg 10,Rm 11N113, Bethesda, MD 20892 USA. NR 62 TC 80 Z9 84 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 JUN 29 PY 2001 VL 276 IS 26 BP 23645 EP 23652 DI 10.1074/jbc.M101031200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 446TW UT WOS:000169531100057 ER PT J AU Coon, SL Weller, JL Korf, HW Namboodiri, MAA Rollag, M Klein, DC AF Coon, SL Weller, JL Korf, HW Namboodiri, MAA Rollag, M Klein, DC TI cAMP regulation of arylalkylamine N-acetyltransferase (AANAT, EC 2.3.1.87) - A new cell line (1E7) provides evidence of intracellular AANAT activation SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID MELATONIN RHYTHM ENZYME; PINEAL-GLAND; PROTEASOMAL PROTEOLYSIS; ANGSTROM RESOLUTION; DISULFIDE EXCHANGE; NOCTURNAL INCREASE; PROTEIN THIOL; CYCLIC-AMP; EXPRESSION; INHIBITION AB Arylalkylamine N-acetyltransferase (serotonin N-acetyltransferase, AANAT, EC 2.3.1.87) is the penultimate enzyme in melatonin synthesis. As described here, a cell line (1E7) expressing human AANAT (hAANAT) has been developed to study the human enzyme. 1E7 hAANAT is detectable in immunoblots as a 23-kDa band and is immunocytochemically visualized in the cytoplasm. The specific concentration of hAANAT in homogenates is comparable to that of the night rat pineal gland. Kinetics of AANAT extracted from 1E7 cells are the same as those of bacterially expressed hAANAT; both preparations of hAANAT are equally sensitive to the inhibitor CoA-S-N-acetyltryptamine. Studies of cAMP regulation indicate that treatment with forskolin, dibutyryl cAMP, isobutylmethylxanthine, or isoproterenol activate cellular hAANAT within intact 1E7 cells similar to8-fold without markedly increasing the abundance of AANAT protein or the activity of AANAT in broken cell preparations; and, that forskolin, isobutylmethylxanthine and isoproterenol elevate cyclic AMP production. These observations extend our understanding of cAMP regulation of AANAT activity, because it is currently thought that this only involves changes in the steady-state levels of AANAT protein. This previously unrecognized switching mechanism could function physiologically to control melatonin production without changing AANAT protein levels. C1 NICHD, Dev Neurobiol Lab, Sect Neuroendocrinol, NIH, Bethesda, MD 20892 USA. Univ Frankfurt, Inst Anat 2, D-60590 Frankfurt, Germany. Uniformed Serv Univ Hlth Sci, Dept Anat Physiol & Genet, Bethesda, MD 20814 USA. RP Klein, DC (reprint author), NICHD, Dev Neurobiol Lab, Sect Neuroendocrinol, NIH, Bldg 49,Rm 6A-80, Bethesda, MD 20892 USA. NR 40 TC 34 Z9 37 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 29 PY 2001 VL 276 IS 26 BP 24097 EP 24107 DI 10.1074/jbc.M011298200 PG 11 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 446TW UT WOS:000169531100116 PM 11313340 ER PT J AU Shi, CS Lee, SB Sinnarajah, S Dessauer, CW Rhee, SG Kehrl, JH AF Shi, CS Lee, SB Sinnarajah, S Dessauer, CW Rhee, SG Kehrl, JH TI Regulator of G-protein signaling 3 (RGS3) inhibits G beta(1)gamma(2)-induced inositol phosphate production, mitogen-activated protein kinase activation, and Akt activation SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID BETA-GAMMA-SUBUNITS; HETEROTRIMERIC G-PROTEINS; ALPHA-SUBUNIT; B-LYMPHOCYTES; BINDING; RECEPTORS; PHOSDUCIN; CHEMOTAXIS; PATHWAY; COMPLEX AB Regulator of G-protein signaling 3 (RGS3) enhances the intrinsic rate at which G alpha (i) and G alpha (q) hydrolyze GTP to GDP, thereby limiting the duration in which GTP-G alpha (i) and GTP-G alpha (q) can activate effecters. Since GDP-G alpha subunits rapidly combine with free G beta gamma subunits to reform inactive heterotrimeric G-proteins, RGS3 and other RGS proteins may also reduce the amount of G beta gamma subunits available for effector interactions. Although RGS6, RGS7, and RGS11 bind GP, in the absence of a Gy subunit, RGS proteins are not known to directly influence G beta gamma signaling. Here we show that RGS3 binds G beta (1)gamma (2) subunits and limits their ability to trigger the production of inositol phosphates and the activation of Akt and mitogen-activated protein kinase. Go-expression of RGS3 with G beta (1)beta (2) inhibits G beta (1)gamma (2) induced inositol phosphate production and Akt activation in COS-7 cells and mitogen-activated protein kinase activation in HEK 293 cells. The inhibition of G beta (1)gamma (2) signaling does not require an intact RGS domain but depends upon two regions in RGS3 located between acids 313 and 390 and between 391 and 458. Several other RGS proteins do not affect G beta (1)gamma (2) signaling in these assays. Consistent with the in vivo results, RGS3 inhibits G beta gamma -mediated activation of phospholipase C beta in vitro. Thus, RGS3 may limit G beta gamma signaling not only by virtue of its GTPase-activating protein activity for G alpha subunits, but also by directly interfering with the activation of effecters. C1 NIAID, Immunoregulat Lab, Cell Mol Immunol Sect B, NIH, Bethesda, MD 20892 USA. Univ Texas, Sch Med, Dept Integrat BIol & Pharmacol, Houston, TX 77030 USA. NHLBI, Lab Cell Signaling, NIH, Bethesda, MD 20892 USA. RP Kehrl, JH (reprint author), NIAID, Immunoregulat Lab, Cell Mol Immunol Sect B, NIH, Bldg 10,Rm 11B08, Bethesda, MD 20892 USA. FU NIGMS NIH HHS [R01 GM060419, R01 GM060419-02, GM 60419] NR 42 TC 34 Z9 36 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 29 PY 2001 VL 276 IS 26 BP 24293 EP 24300 DI 10.1074/jbc.M100089200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 446TW UT WOS:000169531100140 PM 11294858 ER PT J AU Yam, A Hyun, T Li, WQ AF Yam, A Hyun, T Li, WQ TI Characterization of insulin-like growth factor I (IGF-I) receptor mutants for their effects on IGF-I- and interleukin 4-mediated DNA synthesis of 32D cells SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID TYROSINE KINASE DOMAIN; TRANSFORMING ACTIVITIES; TUMOR-GROWTH; MITOGENESIS; SUBSTRATE-1; MUTATIONS; DIFFERENTIATION; PROLIFERATION; ACTIVATION; STATS AB Recently we demonstrated that overexpression of the wild type insulin-like growth factor I receptor (IGF-IRWT) in 32D myeloid progenitor cells led to cell proliferation in response to interleukin 4 (IL-4) as well as insulin-like growth factor I (IGF-I) in the absence of insulin receptor substrate expression (Soon, L,, Flechner, L., Gutkind, J, S., Wang, L, H,, Baserga, R,, Pierce, J, H,, and Li, W. (1999)Mol. Cell. Biol. 19, 3816-3828), To understand the structural importance of insulin-like growth factor I receptor (IGF-IR) in mediating IL-4- and IGF-I-induced DNA synthesis, we transfected various mutants of IGF-IR to 32D cells. Our results show that most mutants, including Y1250F, Y1251F, Y1250F/Y1251F, S1280A/S1281A/S1282A/S1283A, Y1316F, and 1245d, still retained mitogenic response toward IGF-I or IL-4. However, the Y950F, Y1131F, and Y1135F mutants were not able to respond to either ligand, The H1293F/K1294R and 1293d mutants reduced response toward IGF-I but not to IL-4, Phosphorylation of She was greatly reduced in those three mutants that lost mitogenic response. The MAPK activity was much lower in Y1131F and Y1135F mutants, indicating the importance of the Shc/MAPK pathway in IGF-I-induced mitogenesis. Importantly, the synergistic effect of these two factors on DNA synthesis was not affected in cells expressing most of the mutants, even in those three that had lower mitogenic response toward a single ligand, These results suggest that an unidentified pathway(s) may be induced upon co-addition of IGF-I and IL-4 that sustains the intact mitogenesis. C1 Georgetown Univ, Med Ctr, Dept Oncol, Lombardi Canc Ctr, Washington, DC 20007 USA. NCI, Cellular & Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Li, WQ (reprint author), Georgetown Univ, Med Ctr, Dept Oncol, Lombardi Canc Ctr, New Res Bldg,E407,3970 Reservoir Rd, Washington, DC 20007 USA. FU NCI NIH HHS [2P30-CA-51008] NR 27 TC 5 Z9 5 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 29 PY 2001 VL 276 IS 26 BP 24409 EP 24413 DI 10.1074/jbc.M102358200 PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 446TW UT WOS:000169531100156 PM 11323432 ER PT J AU Park, K Chattoraj, DK AF Park, K Chattoraj, DK TI DnaA boxes in the P1 plasmid origin: The effect of their position on the directionality of replication and plasmid copy number SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE origin of replication; plasmid P1; DnaA box; origin-opening; direction of replication ID ESCHERICHIA-COLI CHROMOSOME; OPEN-COMPLEX-FORMATION; INITIATION SITES; MINIMAL REPLICON; IN-VITRO; PROTEIN; TRANSCRIPTION; PSC101; BINDING; R6K AB The DnaA protein is essential for initiation of DNA replication in a wide variety of bacterial and plasmid replicons. The replication origin in these replicons invariably contains specific binding sites for the protein, called DnaA boxes. Plasmid P1 contains a set of DnaA boxes at each end of its origin but can function with either one of the sets. Here we report that the location of origin-opening, initiation site of replication forks and directionality of replication do not change whether the boxes are present at both or at one of the ends of the origin. Replication was bidirectional in all cases. These results imply that DnaA functions similarly from the two ends of the origin. However, origins with DnaA boxes proximal to the origin-opening location opened more efficiently and maintained plasmids at higher copy numbers. Origins with the distal set were inactive unless the adjacent P1 DNA sequences beyond the boxes were included. At either end, phasing of the boxes with respect to the remainder of the origin influenced the copy number. Thus, although the boxes can be at either end, their precise context is critical for efficient origin function. (C) 2001 Academic Press. C1 NCI, Biochem Lab, Canc Res Ctr, NIH, Bethesda, MD 20892 USA. RP Chattoraj, DK (reprint author), NCI, Biochem Lab, Canc Res Ctr, NIH, Bethesda, MD 20892 USA. NR 59 TC 16 Z9 16 U1 0 U2 0 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-2836 J9 J MOL BIOL JI J. Mol. Biol. PD JUN 29 PY 2001 VL 310 IS 1 BP 69 EP 81 DI 10.1006/jmbi.2001.4741 PG 13 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 448JN UT WOS:000169623900005 PM 11419937 ER PT J AU Ramakrishnan, B Qasba, PK AF Ramakrishnan, B Qasba, PK TI Crystal structure of lactose synthase reveals a large conformational change in its catalytic component, the beta 1,4-galactosyltransferase-1 SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE lactose synthase; crystal structure; substrate binding; conformational changes; modulation ID ALPHA-LACTALBUMIN; ESCHERICHIA-COLI; GALACTOSYLTRANSFERASE; EXPRESSION; SYNTHETASE; MECHANISM; LYSOZYME; CLONING; FAMILY; SITE AB The lactose synthase (LS) enzyme is a 1:1 complex of a catalytic component, beta1,4-galactosyltransferse (beta 4Gal-T1) and a regulatory component, a-lactalbumin (LA), a mammary gland-specific protein. LA promotes the binding of glucose (Glc) to beta 4Gal-T1, thereby altering its sugar acceptor specificity from N-acetylglucosamine (GlcNAc) to glucose, which enables LS to synthesize lactose, the major carbohydrate component of milk. The crystal structures of LS bound with various substrates were solved at 2 Angstrom resolution. These structures reveal that upon substrate binding to beta 4Gal-T1, a large conformational change occurs in the region comprising residues 345 to 365. This repositions His347 in such a way that it can participate in the coordination of a metal ion, and creates a sugar and LA-binding site. At the sugar-acceptor binding site, a hydrophobic N-acetyl group-binding pocket is found, formed by residues Arg359, Phe360 and Ile363. In the Glc-bound structure, this hydrophobic pocket is absent. For the binding of Glc to LS, a reorientation of the Arg359 sidechain occurs, which blocks the hydrophobic pocket and maximizes the interactions with the Glc molecule. Thus, the role of LA is to hold Glc by hydrogen bonding with the O-1 hydroxyl group in the acceptor-binding site on beta 4Gal-T1, while the N-acetyl group-binding pocket in beta 4Gal-T1 adjusts to maximize the interactions with the Glc molecule. This study provides details of a structural basis for the partially ordered kinetic mechanism proposed for lactose synthase. C1 NCI, CCR, Struct Glycobiol Sect, Frederick, MD 21702 USA. NCI, Intramural Res Support Program, SAIC, Lab Expt & Computat Biol,CCR, Frederick, MD 21702 USA. RP Qasba, PK (reprint author), NCI, CCR, Struct Glycobiol Sect, Frederick, MD 21702 USA. FU NCI NIH HHS [N01-CO-56000] NR 35 TC 128 Z9 133 U1 0 U2 6 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 JUN 29 PY 2001 VL 310 IS 1 BP 205 EP 218 DI 10.1006/jmbi.2001.4757 PG 14 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 448JN UT WOS:000169623900015 PM 11419947 ER PT J AU Eisenmesser, EZ Horita, DA Altieri, AS Byrd, RA AF Eisenmesser, EZ Horita, DA Altieri, AS Byrd, RA TI Solution structure of interleukin-13 and insights into receptor engagement SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE cytokines; interleukin-13; IL-13; receptor; protein structure ID RECOMBINANT HUMAN INTERLEUKIN-13; 3-DIMENSIONAL SOLUTION STRUCTURE; COLONY-STIMULATING FACTOR; IL-4 RECEPTOR; ALPHA-CHAIN; PSEUDOMONAS EXOTOXIN; CRYSTAL-STRUCTURE; CHEMICAL-SHIFT; PROTEIN; BINDING AB The complex and interrelated function of the interleukin cytokines relies on a range of pro-inflammatory and an ti-inflammatory immune responses mediated by an array of receptors, and there is considerable cross-reactivity for related cytokines. Recent findings continue to elucidate the expression patterns of interleukin receptors associated with a range of diseases, including cancer. We report here the first experimentally determined high-resolution structure of human interleukin-13 (IL-13). The experimental structure is significantly different from an earlier homology model, which could have led to improper estimation of receptor interaction surfaces and design of mutational experiments. Similarities between the presented IL-13 structure and the homogolous interleukin-4 (IL-4) are discussed. Additionally, mutation data for IL-4 and IL-13 are analyzed and combined with a detailed structural analysis of the IL-4/IL4R alpha interface that leads us to postulate interactions at the IL-13/receptor interface. The structural comparison is used to interpret the different affinities for various receptors and establishes the basis for further mutational experiments and antagonist design. C1 Natl Canc Inst, Macromol NMR Sect, Structural Biophys Lab, Frederick, MD 21702 USA. RP Byrd, RA (reprint author), Natl Canc Inst, Macromol NMR Sect, Structural Biophys Lab, Frederick, MD 21702 USA. RI Byrd, R. Andrew/F-8042-2015; OI Byrd, R. Andrew/0000-0003-3625-4232; Horita, David/0000-0002-9563-107X NR 53 TC 30 Z9 33 U1 0 U2 7 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 JUN 29 PY 2001 VL 310 IS 1 BP 231 EP 241 DI 10.1006/jmbi.2001.4765 PG 11 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 448JN UT WOS:000169623900017 PM 11419949 ER PT J AU Dolensky, B Kirk, KL AF Dolensky, B Kirk, KL TI New building blocks for fluorinated imidazole derivatives: Preparation of beta-fluoro- and beta,beta-difluorohistamine SO JOURNAL OF ORGANIC CHEMISTRY LA English DT Article ID HYDROGEN-FLUORIDE; INHIBITORS; AGENTS; ACIDS AB We demonstrate that "FBr" addition to 1-trityl-4-vinyl-1H-imidazole (7) provides a convenient route to side-chain-fluorinated histamines. Thus, addition of "FBr" to the double bond of 7 occurs with Markovnikov regioselectivity to produce 4-(2-bromo-1-fluoroethyl)-1-trityl-1H-imidazole (8). Substitution with azide, reduction, and removal of the trityl group provide beta -fluorohistamine (1) as the dihydrochloride. Elimination of HBr from 8 followed by a second addition of "FBr" gives 4-(2-bromo-1,1-difluoroethyl)-1-trityl-1H-imidazole (15). This was similarly converted to beta,beta -difluorohistamine (2) as the dihydrochloride. C1 NIDDKD, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. RP Kirk, KL (reprint author), NIDDKD, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. NR 26 TC 12 Z9 12 U1 2 U2 5 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0022-3263 J9 J ORG CHEM JI J. Org. Chem. PD JUN 29 PY 2001 VL 66 IS 13 BP 4687 EP 4691 DI 10.1021/jo0102415 PG 5 WC Chemistry, Organic SC Chemistry GA 446WG UT WOS:000169536700031 PM 11421793 ER PT J AU Gagneux, P Gonder, MK Goldberg, TL Morin, PA AF Gagneux, P Gonder, MK Goldberg, TL Morin, PA TI Gene flow in wild chimpanzee populations: what genetic data tell us about chimpanzee movement over space and time SO PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY B-BIOLOGICAL SCIENCES LA English DT Article; Proceedings Paper CT Discussion Meeting on the Origins of HIV and the AIDS Epidemic CY SEP 11-12, 2000 CL ROYAL SOCIETY, LONDON, ENGLAND HO ROYAL SOCIETY DE chimpanzee; Pan troglodytes; HIV; SIV; phylogeography ID EAST-AFRICAN CHIMPANZEES; PAN-TROGLODYTES; NUCLEAR-DNA; REPRODUCTIVE STRATEGIES; CAPTURED CHIMPANZEE; DIVERSITY; SEQUENCES; PATERNITY; HIV-1; HAIR AB The isolation of phylogenetically distinct primate immunodeficiency viruses from at least seven wild-born, captive chimpanzees indicates that viruses closely related to HIV-1 may be endemic in some wild chimpanzee populations. The search for the chimpanzee population or populations harbouring these viruses is therefore on. This paper attempts to answer the question of whether or not such populations of chimpanzees are likely to exist at all, and, if so, where they are likely to be found. We summarize what is known about gene flow in wild populations of chimpanzees, both between major phylogeographical subdivisions of the species, and within these subdivisions. Our analysis indicates that hitherto undocumented reproductively isolated chimpanzee populations may in fact exist. This conclusion is based on the observation that, despite limited geographical sampling and limited numbers of genetic loci, conventional notions of the nature and extent of chimpanzee gene flow have recently been substantially revised. Molecular genetic studies using mitochondrial DNA sequences and hypervariable nuclear microsatellite markers have indicated the existence of heretofore undocumented barriers to chimpanzee gene flow. These studies have identified at least one population of chimpanzees genetically distinct enough to be classified into a new subspecies (Pan troglodytes vellerosus). At the same time, they have called into question the long-accepted genetic distinction between eastern chimpanzees (Pan troglodytes schweinfurthii) and western equatorial chimpanzees (Pan troglodytes troglodytes). The same studies have further indicated that gene flow between local populations is more extensive than was previously thought, and follows patterns sometimes inconsistent with those documented through direct behavioural observation. Given the apparently incomplete nature of the current understanding of chimpanzee gene flow in equatorial Africa, it seems reasonable to speculate that a chimpanzee population or populations may exist which both harbour the putative HIV-1 ancestor, and which have remained reproductively isolated from other chimpanzee populations over the time-scale relevant to the evolution of the SIVcpz-HIV-1 complex of viruses. Continued extensive sampling of wild chimpanzee populations, both for their genes and their viruses, should be performed quickly considering the high probability of extinction that many wild chimpanzee populations face today. The history of human-chimpanzee contacts is discussed. C1 Univ Calif San Diego, Glycobiol Res & Training Ctr, La Jolla, CA 92093 USA. NIDDKD, Mol & Cellular Biol Lab, Sect Genom Struct & Funct, NIH, Bethesda, MD 20892 USA. Univ Illinois, Coll Vet Med, Dept Pathobiol, Urbana, IL 61820 USA. Max Planck Inst Evolutionary Anthropol, Lab Conservat Genet, D-04103 Leipzig, Germany. RP Gagneux, P (reprint author), Univ Calif San Diego, Glycobiol Res & Training Ctr, Room 1087,9500 Gilman Dr, La Jolla, CA 92093 USA. EM gagneux@biomail.ucsd.edu NR 52 TC 62 Z9 67 U1 1 U2 20 PU ROYAL SOC PI LONDON PA 6-9 CARLTON HOUSE TERRACE, LONDON SW1Y 5AG, ENGLAND SN 0962-8436 J9 PHILOS T R SOC B JI Philos. Trans. R. Soc. B-Biol. Sci. PD JUN 29 PY 2001 VL 356 IS 1410 BP 889 EP 897 DI 10.1098/rstb.2001.0865 PG 9 WC Biology SC Life Sciences & Biomedicine - Other Topics GA 448PW UT WOS:000169636700017 PM 11405936 ER PT J AU Schmidt, B Davis, P Moddemann, D Ohlsson, A Roberts, RS Saigal, S Solimano, A Vincer, M Wright, LL AF Schmidt, B Davis, P Moddemann, D Ohlsson, A Roberts, RS Saigal, S Solimano, A Vincer, M Wright, LL CA Trial Indomethacin Prophylaxis Pre TI Long-term effects of indomethacin prophylaxis in extremely-low-birth-weight infants SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Article ID HEMORRHAGE PREVENTION TRIAL; LOW-DOSE INDOMETHACIN; INTRAVENTRICULAR HEMORRHAGE; PREMATURE-INFANTS; PRETERM INFANTS; HEMODYNAMICS; PREDICTION; CHILDREN; OUTCOMES AB Background: The prophylactic administration of indomethacin reduces the frequency of patent ductus arteriosus and severe intraventricular hemorrhage in very-low-birth-weight infants (those with birth weights below 1500 g). Whether prophylaxis with indomethacin confers any long-term benefits that outweigh the risks of drug-induced reductions in renal, intestinal, and cerebral blood flow is not known. Methods: Soon after they were born, we randomly assigned 1202 infants with birth weights of 500 to 999 g (extremely low birth weight) to receive either indomethacin (0.1 mg per kilogram of body weight) or placebo intravenously once daily for three days. The primary outcome was a composite of death, cerebral palsy, cognitive delay, deafness, and blindness at a corrected age of 18 months. Secondary long-term outcomes were hydrocephalus necessitating the placement of a shunt, seizure disorder, and microcephaly within the same time frame. Secondary short-term outcomes were patent ductus arteriosus, pulmonary hemorrhage, chronic lung disease, ultrasonographic evidence of intracranial abnormalities, necrotizing enterocolitis, and retinopathy. Results: Of the 574 infants with data on the primary outcome who were assigned to prophylaxis with indomethacin, 271 (47 percent) died or survived with impairments, as compared with 261 of the 569 infants (46 percent) assigned to placebo (odds ratio, 1.1; 95 percent confidence interval, 0.8 to 1.4; P=0.61). Indomethacin reduced the incidence of patent ductus arteriosus (24 percent, vs. 50 percent in the placebo group; odds ratio, 0.3; P<0.001) and of severe periventricular and intraventricular hemorrhage (9 percent, vs. 13 percent in the placebo group; odds ratio, 0.6; P=0.02). No other outcomes were altered by the prophylactic administration of indomethacin. Conclusions: In extremely-low-birth-weight infants, prophylaxis with indomethacin does not improve the rate of survival without neurosensory impairment at 18 months, despite the fact that it reduces the frequency of patent ductus arteriosus and severe periventricular and intraventricular hemorrhage. (N Engl J Med 2001;344:1966-72.) Copyright (C) 2001 Massachusetts Medical Society. C1 McMaster Univ, Dept Pediat, Hamilton, ON L8N 3Z5, Canada. McMaster Univ, Dept Clin Epidemiol & Biostat, Hamilton, ON L8N 3Z5, Canada. Royal Womens Hosp, Melbourne, Vic, Australia. Univ Manitoba, Dept Pediat & Child Hlth, Winnipeg, MB R3T 2N2, Canada. Univ Toronto, Dept Pediat, Toronto, ON, Canada. Univ Toronto, Dept Obstet & Gynecol, Toronto, ON, Canada. Univ Toronto, Dept Hlth Adm, Toronto, ON, Canada. Univ British Columbia, Dept Pediat, Vancouver, BC V6T 1W5, Canada. Dalhousie Univ, Dept Pediat, Halifax, NS, Canada. NICHHD, Neonatal Res Network, Bethesda, MD 20892 USA. RP Schmidt, B (reprint author), McMaster Univ, Dept Pediat, HSC 3N11E,1200 Main St W, Hamilton, ON L8N 3Z5, Canada. RI Pritchard, Margo /G-5188-2010 OI Pritchard, Margo /0000-0002-1058-9117 FU NICHD NIH HHS [U10 HD21364, U10 HD21373, U10 HD27851] NR 23 TC 325 Z9 337 U1 0 U2 9 PU MASSACHUSETTS MEDICAL SOC PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD JUN 28 PY 2001 VL 344 IS 26 BP 1966 EP 1972 DI 10.1056/NEJM200106283442602 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 446HQ UT WOS:000169508200002 PM 11430325 ER PT J AU Hedenfalk, I Simon, R Trent, J AF Hedenfalk, I Simon, R Trent, J TI Gene-expression profiles in hereditary breast cancer - Reply SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Letter C1 NIH, Bethesda, MD 20892 USA. RP Hedenfalk, I (reprint author), NIH, Bldg 10, Bethesda, MD 20892 USA. NR 3 TC 1 Z9 1 U1 0 U2 3 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 JUN 28 PY 2001 VL 344 IS 26 BP 2029 EP 2029 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 446HQ UT WOS:000169508200017 ER PT J AU Giannakakou, P Robey, R Fojo, T Blagosklonny, MV AF Giannakakou, P Robey, R Fojo, T Blagosklonny, MV TI Low concentrations of paclitaxel induce cell type-dependent p53, p21 and G1/G2 arrest instead of mitotic arrest: molecular determinants of paclitaxel-induced cytotoxicity SO ONCOGENE LA English DT Article DE paclitaxel; G1 arrest; mitosis; p53; p21; Bcl-2 ID HUMAN CANCER-CELLS; INHIBITOR-INDUCED APOPTOSIS; TUMOR-SUPPRESSOR P53; SV40-TRANSFORMED CELLS; ANTICANCER AGENTS; G(1) ARREST; G2/M ARREST; HUMAN HEAD; TAXOL; OVARIAN AB Paclitaxel (PTX), a microtubule-active agent, blocks cell proliferation by inhibiting mitotic progression leading to mitotic and postmitotic arrest and cell death. Here we demonstrate for the first time that very low concentrations of PTX (3-6 nM) can completely inhibit cell proliferation without arresting cells at mitosis, At these low concentrations that are insufficient to inhibit mitotic progression, PTX induced both p53 and p21 causing CI and G2 arrest in A549. In contrast, low PTX concentrations failed to induce G1 and G2 arrest in A549/E6 cells, that do not express p53, Furthermore, we observed that the levels of p53 and p21 induced by adriamycin and by low concentrations of PTX in A549 cells were comparable. This observation led us to conclude that tow concentrations of PTX can induce p53 and p21 sufficiently to cause G1 and G2, Many other cell lines, including HCT116 cells, do not readily upregulate p53 in response to PTX, and therefore undergo exclusively mitotic and postmitotic arrest after PTX treatment. At low concentrations that do not cause mitotic arrest, PTX did not significantly inhibit proliferation of these cells. In HCT116 cells, loss of p53 (HCT/p53(-/-)) or p21 (HCT/p21(-/-)) affects both Bax and Bcl-2 expression. In cells lacking p53, levels of Bax and p21 were decreased. In cells lacking p21, levels of wt p53 were highly increased to compensate for the loss of p21, This in turn results in upregulation of Bax and downregulation of Bcl-2 resulting in an increase of the apoptotic Bax/Bcl2 ratio consistent with increased sensitivity of these cells to apoptotic stimuli, High levels of p53 and Bax/Bcl-2 ratio can also explain why loss of p21 is rarely found in human cancer. C1 NCI, Med Branch, NIH, Bethesda, MD 20892 USA. Emory Univ, Atlanta, GA 30322 USA. RP Blagosklonny, MV (reprint author), NCI, Med Branch, NIH, Bldg 10,R 12N226, Bethesda, MD 20892 USA. OI Giannakakou, Paraskevi/0000-0001-7378-262X NR 56 TC 146 Z9 162 U1 0 U2 8 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD JUN 28 PY 2001 VL 20 IS 29 BP 3806 EP 3813 DI 10.1038/sj.onc.1204487 PG 8 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 446CE UT WOS:000169494700006 PM 11439344 ER PT J AU Frisch, M Biggar, RJ Engels, EA Goedert, JJ AF Frisch, M Biggar, RJ Engels, EA Goedert, JJ TI Breast cancer in women with HIV/AIDS - Reply SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Letter C1 State Serum Inst, Danish Epidemiol Sci Ctr, Copenhagen, Denmark. NCI, Viral Epidemiol Branch, Div Epidemiol & Genet, Bethesda, MD 20892 USA. RP Frisch, M (reprint author), State Serum Inst, Danish Epidemiol Sci Ctr, Copenhagen, Denmark. RI Frisch, Morten/E-9206-2016 OI Frisch, Morten/0000-0002-3864-8860 NR 0 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 JUN 27 PY 2001 VL 285 IS 24 BP 3091 EP 3091 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 444FF UT WOS:000169388900018 ER PT J AU Topol, IA Burt, SK Deretey, E Tang, TH Perczel, A Rashin, A Csizmadia, IG AF Topol, IA Burt, SK Deretey, E Tang, TH Perczel, A Rashin, A Csizmadia, IG TI alpha- and 3(10)-helix interconversion: A quantum-chemical study on polyalanine systems in the gas phase and in aqueous solvent SO JOURNAL OF THE AMERICAN CHEMICAL SOCIETY LA English DT Article ID ALANINE-BASED PEPTIDES; CONTINUUM DIELECTRIC THEORY; ELECTRON-SPIN-RESONANCE; MOLECULAR-DYNAMICS; RICH PEPTIDES; AMINO-ACIDS; EQUILIBRIUM; TRANSITION; STABILITY; WATER AB Helices are among the predominant secondary structures in globular proteins. About 90% of the residues in them are found to be in the alpha -helical conformation, and another 10% in the 3(10) conformation. There is a standing controversy between experimental and some theoretical results, and controversy among theoretical results concerning the predominance of each conformation, in particular, helices, We address this controversy by ab initio Hartree-Fock and density functional theory studies of helices with different lengths in a vacuum and in the aqueous phase. Our results show that (1) in a vacuum, all oligo(Ala) helices of 4-10 residues adopt the 3(10) - conformation; (2) in aqueous solution, the 6-10 residue peptides adopt the alpha -helical conformation; (3) there might be two intermediates between these helical conformers allowing for their interconversion. The relevance of these results to the structure and folding of proteins is discussed. C1 NCI, Adv Biomed Comp Ctr, SAIC Frederick, Frederick, MD 21702 USA. Univ Toronto, Dept Chem, Toronto, ON M5S 3H6, Canada. Eotvos Lorand Univ, Inst Organ Chem, H-1117 Budapest, Hungary. BioChemComp Inc, Teaneck, NJ 07666 USA. RP NCI, Adv Biomed Comp Ctr, SAIC Frederick, POB B, Frederick, MD 21702 USA. EM topol@ncifcrf.gov; burt@ncifcrf.gov; ederetey@alchemy.chem.utoronto.ca; ttang@alchemy.chem.utoronto.ca; aperczel@biotech.ox.ac.uk; rashin@idt.net; icsizmad@alchemy.chem.utoronto.ca FU NCI NIH HHS [N01-CO-56000] NR 40 TC 80 Z9 81 U1 3 U2 10 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 JUN 27 PY 2001 VL 123 IS 25 BP 6054 EP 6060 DI 10.1021/ja0038934 PG 7 WC Chemistry, Multidisciplinary SC Chemistry GA 446VC UT WOS:000169534000022 PM 11414838 ER PT J AU Ishima, R Petkova, AP Louis, JM Torchia, DA AF Ishima, R Petkova, AP Louis, JM Torchia, DA TI Comparison of methyl rotation axis order parameters derived from model-free analyses of H-2 and C-13 longitudinal and transverse relaxation rates measured in the same protein sample SO JOURNAL OF THE AMERICAN CHEMICAL SOCIETY LA English DT Article ID MAGNETIC-RESONANCE RELAXATION; NUCLEAR-SPIN RELAXATION; NMR RELAXATION; SIDE-CHAINS; DYNAMICS; SPECTROSCOPY; N-15; MACROMOLECULES; ANISOTROPY; DIFFUSION AB Recombinant HIV-1 protease was obtained from bacteria grown on a 98% D2O medium containing 3-C-13 pyruvic acid as the sole source of C-13 and H-1, The purified protein is highly deuterated at non-methyl carbons, but contains significant populations of (CHD2)-C-13 and (CH2D)-C-13 methyl isotopomers. This pattern of isotope labeling permitted measurements of H-1 and C-13 relaxation rates of (CHD2)-C-13 isotopomers and H-2 (D) relaxation rates of (CH2D)-C-13 isotopomers using a single sample. The order parameters S-axis(2), which characterize the motions of the methyl rotation axes, were derived from model-free analyses of R-1 and R-2 data sets measured for C-13 and H-2 spins. Our primary goal was to compare the S-axis(2) values derived from the two independent types of data sets to test our understanding of the relaxation mechanisms involved. However, S-axis(2) values derived from the analyses depend strongly on the geometry of the methyl group, the sizes of the quadrupolar and dipolar couplings, and the effects of bond vibrations and librations on these couplings. Therefore uncertainties in these basic physical parameters complicate comparison of the order parameters. This problem was circumvented by using an experimental relationship, between the methyl quadrupolar, C-13-C-13 and C-13-H-1 dipolar couplings, derived from independent measurements of residual static couplings of weakly aligned proteins by Ottiger and Bar (J. Am. Chem. Soc. 1999, 121, 4690-4695) and Mittermaier and Kay (J. Am. Chem. Sec. 1999, 121, 10608-10613). This approach placed a tight experimental restraint on the values of the H-2 quadrupolar and C-13-H-1 dipolar interactions and greatly facilitated the accurate comparison of the relative values of the order parameters. When applied to our data this approach yielded satisfactory agreement between the S-axis(2) values derived from the C-13 and H-2 data sets. C1 Natl Inst Dent & Craniofacial Res, Struct Mol Biol Unit, NIH, Bethesda, MD 20892 USA. NIDDKD, Chem Phys Lab, NIH, Bethesda, MD 20892 USA. RP Torchia, DA (reprint author), Natl Inst Dent & Craniofacial Res, Struct Mol Biol Unit, NIH, Bethesda, MD 20892 USA. NR 40 TC 69 Z9 70 U1 2 U2 10 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 JUN 27 PY 2001 VL 123 IS 25 BP 6164 EP 6171 DI 10.1021/ja0104711 PG 8 WC Chemistry, Multidisciplinary SC Chemistry GA 446VC UT WOS:000169534000035 PM 11414851 ER PT J AU Ferrari, D Yang, LH Miles, EW Dunn, MF AF Ferrari, D Yang, LH Miles, EW Dunn, MF TI beta D305A mutant of tryptophan synthase shows strongly perturbed allosteric regulation and substrate specificity SO BIOCHEMISTRY LA English DT Article ID ULTRAVIOLET-VISIBLE SPECTROSCOPY; INDUCED CONFORMATIONAL-CHANGES; LIVER ALCOHOL-DEHYDROGENASE; MONOVALENT CATION ACTION; SERINE O-SULFATE; ESCHERICHIA-COLI; BIENZYME COMPLEX; ALPHA-SUBUNIT; SALMONELLA-TYPHIMURIUM; ALPHA(2)BETA(2) COMPLEX AB Substrate channeling in the tryptophan synthase bienzyme is regulated by allosteric interactions. Allosteric signals are transmitted via a scaffolding of structural elements that includes a monovalent cation-binding site and salt-bridging interactions between the side chains of beta Asp 305, beta Arg 141, beta Lys 167, and alpha Asp 56 that appear to modulate the interconversion between open and closed conformations. beta Asp 305 also interacts with the hydroxyl group of the substrate L-Ser in some structures. One possible functional role for beta Asp 305 is to ensure the allosteric transmission that triggers the switching of alpha beta -dimeric units between open and closed conformations of low and high activity. This work shows that substitution of beta Asp 305 with Ala (beta D305A) decreases the affinity of the beta -site for the substrate L-Ser, destabilizes the enzyme-bound alpha -aminoacrylate, E(A-A), and quinonoid species, E(Q), and changes the nucleophile specificity of the beta -reaction. The altered specificity provides a biosynthetic route for new L-amino acids reaction with L-Ser relative to the wild-type enzyme. The formation of pyruvate is strongly inhibited by the binding of benzimidazole to E(A-A). Upon reaction with L-Ser and in the presence of the alpha -site substrate analogue, alpha -glycerol phosphate, the Na+ form of beta D305A undergoes inactivation via reaction of nascent alpha -aminoacrylate with bound PLP. This work establishes important roles for beta Asp 305 both in the conformational change between open and closed states that takes place at the beta -site during the formation of the E(A-A) and in substrate binding and recognition. C1 Univ Calif Riverside, Dept Biochem, Riverside, CA 92521 USA. NIDDK, NIH, Lab Biochem & Genet, Bethesda, MD 20892 USA. RP Dunn, MF (reprint author), Univ Calif Riverside, Dept Biochem, Riverside, CA 92521 USA. EM dunn@ucrac1.ucr.edu FU NIGMS NIH HHS [GM55749] NR 49 TC 31 Z9 31 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 JUN 26 PY 2001 VL 40 IS 25 BP 7421 EP 7432 DI 10.1021/bi002892l PG 12 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 446WC UT WOS:000169536300007 PM 11412095 ER PT J AU Aszalos, A Ladanyi, A Bocsi, J Szende, B AF Aszalos, A Ladanyi, A Bocsi, J Szende, B TI Induction of apoptosis in MDR1 expressing cells by daunorubicin with combinations of suboptimal concentrations of P-glycoprotein modulators SO CANCER LETTERS LA English DT Article DE multidrug resistance; Pgp inhibitors; daunorubicin; apoptosis ID CREMOPHOR-EL; IN-VITRO; RESISTANCE; PROLIFERATION; VERAPAMIL; BLOCKERS; TUMOR AB The application of most agents with the capacity to reverse multidrug resistance (MDR) via modulation of the multidrug transporter P-glycoprotein (Pgp) was shown to be associated with toxic side-effects. For this reason, we have investigated the effect of combinations of suboptimal concentrations of Pgp blockers on the induction of apoptosis and growth arrest in daunorubicin (D) treated, MDRI gene transfected cells. We used verapamil, PSC833 and Cremophor EL as Pgp modulators, which affect the function of Pgp by different mechanisms. Treatment of NIH3T3/MDR1 cells with combinations of suboptimal concentrations of Pgp modulators in the presence of D caused apoptosis and G(2) arrest to the same extent as optimal concentrations of singly used blockers. We conclude that combinations of suboptimal concentrations of Pgp modulators may cause effective sensitization of resistant tumor cells, and at the same time, may avoid the frequently observed toxic effects experienced in clinical trials with a single modifier applied at the optimal dose. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved. C1 Natl Inst Oncol, Dept Tumor Progress, H-1122 Budapest, Hungary. NCI, NIH, Bethesda, MD 20892 USA. Semmelweis Univ, Inst Pathol & Expt Canc Res 1, H-1085 Budapest, Hungary. RP Ladanyi, A (reprint author), Natl Inst Oncol, Dept Tumor Progress, Rath Gy U 7-9, H-1122 Budapest, Hungary. EM ladanyi@oncol.hu NR 15 TC 13 Z9 13 U1 0 U2 1 PU ELSEVIER IRELAND LTD PI CLARE PA ELSEVIER HOUSE, BROOKVALE PLAZA, EAST PARK SHANNON, CO, CLARE, 00000, IRELAND SN 0304-3835 J9 CANCER LETT JI Cancer Lett. PD JUN 26 PY 2001 VL 167 IS 2 BP 157 EP 162 DI 10.1016/S0304-3835(01)00488-8 PG 6 WC Oncology SC Oncology GA 437XH UT WOS:000169024300006 PM 11369136 ER PT J AU O'Donnell, CJ Larson, MG Feng, DL Sutherland, PA Lindpaintner, K Myers, RH D'Agostino, RA Levy, D Tofler, GH AF O'Donnell, CJ Larson, MG Feng, DL Sutherland, PA Lindpaintner, K Myers, RH D'Agostino, RA Levy, D Tofler, GH TI Genetic and environmental contributions to platelet aggregation - The Framingham Heart Study SO CIRCULATION LA English DT Article DE platelets; genetics; glycoproteins; fibrinogen ID CORONARY-ARTERY DISEASE; MYOCARDIAL-INFARCTION; ALPHA(2)BETA(1) DENSITY; CARDIOVASCULAR-DISEASE; SYSTEMIC HYPERTENSION; HEMOSTATIC RESPONSES; RISK; POLYMORPHISMS; RECEPTOR; AGGREGABILITY AB Background-Platelet aggregation plays an important role in arterial thrombosis in coronary heart disease, stroke, and peripheral arterial disease. However, the contribution of genetic versus environmental influences on interindividual variation in platelet aggregability is poorly characterized. Methods and Results-We studied the heritability of platelet aggregation responses in 2413 participants in the Framingham Heart Study. The threshold concentrations of epinephrine and ADP required to produce biphasic platelet aggregation and collagen lag time were determined. Mixed-model linear regression was used to calculate correlation coefficients within sibships and within spouse pairs. Variance and covariance component methods were used to estimate the proportion of platelet aggregation attributable to measured covariates versus additive genetic effects. After accounting for environmental covariates, the adjusted sibling correlations for epinephrine, ADP, and collagen lag time were 0.24, 0.22, and 0.31, respectively (P=0.0001 for each). In contrast, adjusted correlations for spouse-pairs were -0.01, 0.05, and -0.02, respectively (all P>0.30). The estimated heritabilities were 0.48, 0.34, and 0.62, respectively. Measured covariates accounted for only 4% to 7% of the overall variance in platelet aggregation, and heritable factors accounted for 20% to 30%. The platelet glycoprotein Illa Pl(A2) polymorphism and the fibrinogen Hind III beta -148 polymorphism contributed <1% to the overall variance. Conclusions-In our large, population-based sample, heritable factors play a major role in determining platelet aggregation, and measured covariates play a lesser role. Future studies are warranted to identify the key genetic variants that regulate platelet function and to lay the groundwork for rational pharmacogenetic approaches. C1 Framingham Heart Study, NHLBI, Framingham, MA 01701 USA. Harvard Univ, Massachusetts Gen Hosp, Sch Med, Dept Med, Boston, MA USA. Harvard Univ, Sch Med, Inst Prevent Cardiovasc Dis, Boston, MA USA. Harvard Univ, Sch Med, Beth Israel Deaconess Med Ctr, Div Cardiol, Boston, MA USA. Harvard Univ, Brigham & Womens Hosp, Sch Med, Dept Med,Cardiovasc Div, Boston, MA 02115 USA. Boston Univ, Sch Med, Dept Neurol, Boston, MA 02118 USA. Boston Univ, Sch Med, Dept Epidemiol & Prevent Med, Boston, MA 02118 USA. Boston Univ, Dept Math, Boston, MA 02215 USA. F Hoffmann La Roche & Co Ltd, CH-4002 Basel, Switzerland. NHLBI, NIH, Bethesda, MD 20892 USA. RP O'Donnell, CJ (reprint author), Framingham Heart Study, NHLBI, 5 Thurber St, Framingham, MA 01701 USA. FU NHLBI NIH HHS [HL NO1-38038] NR 37 TC 137 Z9 139 U1 0 U2 3 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0009-7322 J9 CIRCULATION JI Circulation PD JUN 26 PY 2001 VL 103 IS 25 BP 3051 EP 3056 PG 6 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 449NK UT WOS:000169692200013 PM 11425767 ER PT J AU Ornato, JP Hand, MM AF Ornato, JP Hand, MM TI Warning signs of a heart attack SO CIRCULATION LA English DT Article C1 Virginia Commonwealth Univ, Med Coll Virginia, Dept Emergency Med, Richmond, VA 23298 USA. NHLBI, Natl Heart Attack Alert Program, NIH, Bethesda, MD 20892 USA. RP Ornato, JP (reprint author), Virginia Commonwealth Univ, Med Coll Virginia, Dept Emergency Med, Richmond, VA 23298 USA. NR 0 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 JUN 26 PY 2001 VL 103 IS 25 BP E124 EP E125 PG 2 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 449NK UT WOS:000169692200001 PM 11425785 ER PT J AU Houlden, H Baker, M Morris, HR MacDonald, N Pickering-Brown, S Adamson, J Lees, AJ Rossor, MN Quinn, NP Kertesz, A Khan, MN Hardy, J Lantos, PL George-Hyslop, PS Munoz, DG Mann, D Lang, AE Bergeron, C Bigio, EH Litvan, I Bhatia, KP Dickson, D Wood, NW Hutton, M AF Houlden, H Baker, M Morris, HR MacDonald, N Pickering-Brown, S Adamson, J Lees, AJ Rossor, MN Quinn, NP Kertesz, A Khan, MN Hardy, J Lantos, PL George-Hyslop, PS Munoz, DG Mann, D Lang, AE Bergeron, C Bigio, EH Litvan, I Bhatia, KP Dickson, D Wood, NW Hutton, M TI Corticobasal degeneration and progressive supranuclear palsy share a common tau haplotype SO NEUROLOGY LA English DT Article ID NEURODEGENERATIVE DISEASES; FRONTOTEMPORAL DEMENTIA; CLINICAL-FEATURES; GENETIC-EVIDENCE; MUTATIONS; CHROMOSOME-17; ASSOCIATION; INVOLVEMENT; DIAGNOSIS; ISOFORMS AB Objective: To analyze the association of polymorphisms in the tau gene with pathologically confirmed corticobasal degeneration (CBD). Background: The authors previously described an extended tau haplotype (H1) that covers the human tau gene and is associated with the development of progressive supranuclear palsy (PSP). The authors now extend this analysis to CBD, a neurodegenerative condition with clinical and neuropathologic similarities to PSP. Like PSP, CBD is associated with accumulation of aggregates containing the 4-repeat isoforms of tau. Because of difficulty in diagnosis of CBD, the authors only analyzed cases with pathologically confirmed CBD. Methods: The authors collected 57 unrelated, neuropathologically confirmed cases of CBD. Tau sequencing in these cases failed to show the presence of pathogenic mutations. Polymorphisms that spanned the tau gene were analyzed in all CBD cases and controls. Results: Analyzing tau polymorphisms in CBD cases showed that the frequency of H1 and H1/H1 was significantly increased when analyzing all cases and when separating by country of origin. H1 frequency in all CBD cases was 0.921, compared with a control frequency of 0.766 (X-2 = 9.1, P = 0.00255 [1df], OR 3.56 [8.43 > CI 95% > 1.53]). The H1/H1 frequency was also significantly higher at 0.842 compared with 0.596 in age-matched controls (X-2 = 17.42,p = 0.00016, 2df), OR 3.61 [7.05 > CI 95% > 1.851). Conclusions: The CBD tau association described here suggests that PSP and CBD share a similar cause, although the pathogenic mechanism behind the two diseases leads to a different clinical and pathologic phenotype. C1 Mayo Clin Jacksonville, Jacksonville, FL 32224 USA. Inst Neurol, Clin Neurol & Dementia Res Grp, London WC1N 3BG, England. St Bartholomews Hosp, Dept Gynaecol Oncol, London, England. Univ Manchester, Sch Biol Sci, Manchester M13 9PL, Lancs, England. Univ Manchester, Dept Med, Manchester M13 9PL, Lancs, England. Inst Psychiat, Dept Neuropathol, London SE5 8AF, England. Univ Western Ontario, Dept Neuropathol, London, ON N6A 3K7, Canada. Dept Neurol, Toronto, ON, Canada. Univ Texas, SW Med Sch, Dept Pathol, Dallas, TX 75230 USA. NINDS, NIH, Bethesda, MD 20892 USA. RP Hutton, M (reprint author), Mayo Clin Jacksonville, 4500 San Pablo Rd, Jacksonville, FL 32224 USA. RI Houlden, Henry/C-1532-2008; Lees, Andrew/A-6605-2009; Pickering-Brown, Stuart/D-4008-2009; Morris, Huw/B-8527-2008; turton, miranda/F-4682-2011; Hardy, John/C-2451-2009; Wood, Nicholas/C-2505-2009; OI Houlden, Henry/0000-0002-2866-7777; Pickering-Brown, Stuart/0000-0003-1561-6054; Morris, Huw/0000-0002-5473-3774; Wood, Nicholas/0000-0002-9500-3348; Dickson, Dennis W/0000-0001-7189-7917; Munoz, David/0000-0003-0957-6244; Litvan, Irene/0000-0002-3485-3445 NR 31 TC 248 Z9 251 U1 1 U2 6 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0028-3878 J9 NEUROLOGY JI Neurology PD JUN 26 PY 2001 VL 56 IS 12 BP 1702 EP 1706 PG 5 WC Clinical Neurology SC Neurosciences & Neurology GA 444WZ UT WOS:000169424200016 PM 11425937 ER PT J AU Raben, N Danon, M Lu, N Lee, E Shliselfeld, L Skurat, AV Roach, PJ Lawrence, JC Musumeci, O Shanske, S DiMauro, S Plotz, P AF Raben, N Danon, M Lu, N Lee, E Shliselfeld, L Skurat, AV Roach, PJ Lawrence, JC Musumeci, O Shanske, S DiMauro, S Plotz, P TI Surprises of genetic engineering - A possible model of polyglucosan body disease SO NEUROLOGY LA English DT Article ID GLYCOGEN-BRANCHING ENZYME; MUSCLE PHOSPHOFRUCTOKINASE DEFICIENCY; PROGRESSIVE MYOCLONUS EPILEPSY; ALPHA-GLUCOSIDASE GENE; SKELETAL-MUSCLE; TARGETED DISRUPTION; TRANSGENIC MICE; MUTATIONS; PROTEIN; POLYSACCHARIDE AB Background: The authors previously reported the generation of a knockout mouse model of Pompe disease caused by the inherited deficiency of lysosomal acid alpha-glucosidase (GAA). The disorder in the knockout mice (GAA-/-) resembles the human disease closely, except that the clinical symptoms develop late relative to the lifespan of the animals. In an attempt to accelerate the course of the disease in the knockouts, the authors increased the level of cytoplasmic glycogen by overexpressing glycogen synthase (GSase) or GlutI glucose transporter. Methods: GAA-/- mice were crossed to transgenic mice overexpressing GSase or Glutl in skeletal muscle. Results: Both transgenics on a GAA knockout background (GS/GAA-/- and GlutI/GBA-/-) developed a severe muscle wasting disorder with an early age at onset. This finding, however, is not the major focus of the study. Unexpectedly, the mice bearing the GSase transgene, but not those bearing the GlutI transgene, accumulated structurally abnormal polysaccharide (polyglucosan) similar to that observed in patients with Lafora disease, glycogenosis type IV, and glycogenosis type VII. Ultrastructurally, the periodic acid-Schiff (PAS)-positive polysaccharide inclusions were composed of short, amorphous, irregular branching filaments indistinguishable from classic polyglucosan bodies. The authors show here that increased level of GSase in the presence of normal glycogen branching enzyme (GBE) activity leads to polyglucosan accumulation. The authors have further shown that inactivation of lysosomal acid alpha-glucosidase in the knockout mice does not contribute to the process of polyglucosan formation. Conclusions: An imbalance between GSase and GEE activities is proposed as the mechanism involved in the production of polyglucosan bodies. The authors may have inadvertently created a "muscle polyglucosan disease" by simulating the mechanism for polyglucosan formation. C1 NIAMSD, NIH, Bethesda, MD 20892 USA. NYU, Sch Med, Dept Neurol & Pathol, New York, NY USA. NYU, Sch Med, Dept Neurol, New York, NY USA. Indiana Univ, Sch Med, Dept Biochem, Indianapolis, IN 46202 USA. Univ Virginia, Dept Pharmacol, Charlottesville, VA 22908 USA. Columbia Univ Coll Phys & Surg, Dept Neurol, New York, NY 10032 USA. RP Raben, N (reprint author), NIAMS, NIH, Bldg 10-9N244,9000 Rockville Pike, Bethesda, MD 20892 USA. FU NIDDK NIH HHS [R01 DK28312]; NINDS NIH HHS [NS11766] NR 35 TC 54 Z9 54 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0028-3878 J9 NEUROLOGY JI Neurology PD JUN 26 PY 2001 VL 56 IS 12 BP 1739 EP 1745 PG 7 WC Clinical Neurology SC Neurosciences & Neurology GA 444WZ UT WOS:000169424200022 PM 11425943 ER PT J AU Bollmann, R Bollmann, M Henson, DE Bodo, M AF Bollmann, R Bollmann, M Henson, DE Bodo, M TI DNA cytometry confirms the utility of the Bethesda System for the classification of Papanicolaou smears SO CANCER CYTOPATHOLOGY LA English DT Article DE Bethesda System; DNA cytometry; Munich II Nomenclature; Papanicolaou smear; uterine cervical carcinoma ID CERVICAL INTRAEPITHELIAL NEOPLASIA; HUMAN-PAPILLOMAVIRUS TYPE-16; UTERINE CERVIX; NUCLEAR-DNA; PROSPECTIVE MALIGNANCY; IMAGE-CYTOMETRY; CHROMATIN COMPACTNESS; EPITHELIAL NEOPLASIA; BORDERLINE LESIONS; MODEL FILMS AB BACKGROUND. Developed in 1989, the Bethesda System has largely replaced previous classifications of Papanicolaou (Pap) smears front the uterine cervix. The system is binary, dividing smears into two groups - low-grade, squamous, epithelial lesions (LSIL) or high-grade, squamous, epithelial lesions (HSIL). A third category, atypical squamous cells of undetermined significance (ASCUS), is used to classify minimal cellular changes that do not satisfy the criteria for the low- or high-grade categories. This study was designed to confirm the utility of this binary division and to compare the results with another classification system (the Munich II Nomenclature) that is not binary but contains three divisions or grades for dysplasia - low, intermediate, and high. METHODS, Pap smears were obtained from 593 women with a cytologic diagnosis of dysplasia based on the Munich System. Smears were then classified by the Bethesda System into LSIL or HSIL. Patients were followed for 2 years either with biopsy or repeat cytology. The initial smears were restained by the Feulgen method, and ploidy was evaluated by interactive DNA cytometry. RESULTS. Of 241 cases of LSIL, 39% were diploid, 57% polyploid, and 4% aneuploid, Of 352 cases classified HSIL, 4% were diploid, 17% polyploid, and 79% aneuploid. After 2 years of follow-up, 2 of 108 patients who were biopsied and who were originally classified as diploid progressed to cervical intraepithelial neoplasia/ carcinoma in situ (CIN/CIS) whereas 109 of 217 patients who were aneuploid and biopsied were found to have CINIII/CIS. CONCLUSIONS, The two divisions of the Bethesda System, LSIL and HSIL, correlated with ploidy as evaluated by cytometry. Aneuploidy was found to be useful to separate cases of HSIL from those of LSIL as defined in the Bethesda System. Because of the binary division, use of a system with three divisions for dysplasia, such as the Munich II Nomenclature, creates a therapeutic dilemma because a single diagnostic category (usually the intermediate grade) may contain both self-limiting and progressive lesions. DNA cytometry of Pap smears was found to be useful as a routine procedure. Cancer (Cancer Cytopathol) 2001;93:222-228. (C) 2001 American Cancer Society. C1 NCI, Canc Biomarkers Res Grp, Div Canc Prevent, Bethesda, MD 20892 USA. Univ Bonn, Inst Pathol, D-5300 Bonn, Germany. St Johns Hosp, Dept Pathol, Budapest, Hungary. RP Henson, DE (reprint author), NCI, Canc Biomarkers Res Grp, Div Canc Prevent, EPN Bldg,Room 305,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 84 TC 23 Z9 25 U1 1 U2 2 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 JUN 25 PY 2001 VL 93 IS 3 BP 222 EP 228 DI 10.1002/cncr.9033 PG 7 WC Oncology; Pathology SC Oncology; Pathology GA 442LD UT WOS:000169286500009 PM 11391611 ER PT J AU Marshall, JG Booth, JW Stambolic, V Mak, T Balla, T Schreiber, AD Meyer, T Grinstein, S AF Marshall, JG Booth, JW Stambolic, V Mak, T Balla, T Schreiber, AD Meyer, T Grinstein, S TI Restricted accumulation of phosphatidylinositol 3-kinase products in a plasmalemmal subdomain during Fc gamma receptor-mediated phagocytosis SO JOURNAL OF CELL BIOLOGY LA English DT Article DE Fc gamma receptors; PH domain; phosphoinositide; lipid mobility; FRAP ID PROTEIN-KINASE-C; LIVING CELLS; PHOSPHOINOSITIDE 3-KINASE; SIGNAL-TRANSDUCTION; MACROPHAGES; ACTIVATION; PHAGOSOMES; MEMBRANE; CHOLESTEROL; INVOLVEMENT AB Phagocytosis is a highly localized and rapid event, requiring the generation of spatially and temporally restricted signals. Because phosphatidylinositol 3-kinase (PI3K) plays an important role in the innate immune response, we studied the generation and distribution of 3' phosphoinositides (3'PIs) in macrophages during the course of phagocytosis, The presence of 3'PI was monitored noninvasively in cells transfected with chimeras of green fluorescent protein and the pleckstrin homology domain of either Akt, Btk, or Gab1, Although virtually undetectable in unstimulated cells, 3'PI rapidly accumulated at sites: of phagocytosis. This accumulation was sharply restricted to the phagosomal cup, with little 3'PI detectable in the immediately adjacent areas of the plasmalemma. Measurements of fluorescence recovery after photobleaching were made to estimate the mobility of lipids in the cytosolic monolayer of the phagosomal membrane. Stimulation of phagocytic receptors induced a marked reduction of lipid mobility that likely contributes to the restricted distribution of 3'PI at the cup. 3'PI accumulation during phagocytosis was transient, terminating shortly after sealing of the phagosomal vacuole. Two factors contribute to the rapid disappearance of 3'PI: the dissociation of the type I PI3K from the phagosomal membrane and the persistent accumulation of phosphoinositide phosphatases. C1 Hosp Sick Children, Div Cell Biol, Toronto, ON M5G 1X8, Canada. Amgen Inst, Toronto, ON M5G 2C1, Canada. NICHHD, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. Univ Penn, Sch Med, Dept Med, Philadelphia, PA 19104 USA. Stanford Univ, Dept Mol Pharmacol, Stanford, CA 94305 USA. RP Marshall, JG (reprint author), Hosp Sick Children, Div Cell Biol, 555 Univ Ave, Toronto, ON M5G 1X8, Canada. RI Marshall, John /E-8974-2012; OI Marshall, John /0000-0002-7124-1448; Balla, Tamas/0000-0002-9077-3335 FU NHLBI NIH HHS [HL28207, R01 HL028207] NR 35 TC 194 Z9 196 U1 0 U2 4 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD JUN 25 PY 2001 VL 153 IS 7 BP 1369 EP 1380 DI 10.1083/jcb.153.7.1369 PG 12 WC Cell Biology SC Cell Biology GA 447QJ UT WOS:000169583700004 PM 11425868 ER PT J AU Jung, G Remmert, K Wu, XF Volosky, JM Hammer, JA AF Jung, G Remmert, K Wu, XF Volosky, JM Hammer, JA TI The Dictyostelium CARMIL protein links capping protein and the Arp2/3 complex to type I myosins through their SH3 domains SO JOURNAL OF CELL BIOLOGY LA English DT Article DE myosin I; Arp2/3 complex; capping protein; leucine-rich repeats; Dictyostelium ID WISKOTT-ALDRICH-SYNDROME; ACTIN-BASED MOTILITY; LEUCINE-RICH REPEATS; WASP-RELATED PROTEIN; HEAVY-CHAIN ISOFORM; N-WASP; ACANTHAMOEBA-CASTELLANII; WASP/SCAR PROTEINS; F-ACTIN; YEAST AB Fusion proteins containing the Src homology (SH)3 domains of Dictyostelium myosin IB (myoB) and IC (myoC) bind a 116-kD protein (p116), plus nine other proteins identified as the seven member Arp2/3 complex, and the alpha and beta subunits of capping protein. Immunoprecipitation reactions indicate that myoB and myoC form a complex with p116, Arp2/3, and capping protein in vivo, that the myosins bind to p116 through their SH3 domains, and that capping protein and the Arp2/3 complex in turn bind to p116. Cloning of p116 reveals a protein dominated by leucine-rich repeats and proline-rich sequences, and indicates that it is a homologue of Acan 125. Studies using p116 fusion proteins confirm the location of the myosin I SH3 domain binding site, implicate NH2-terminal sequences in binding capping protein, and show that a region containing a short sequence found in several G-actin binding proteins, as well as an acidic stretch, can activate Arp2/3-dependent actin nucleation. p116 localizes along with the Arp2/3 complex, myoB, and myoC in dynamic astin-rich cellular extensions, including the leading edge of cells undergoing chemotactic migration, and dorsal, cup-like, macropinocytic extensions. Cells lacking p116 exhibit a striking defect in the formation of these macropinocytic structures, a concomitant reduction in the rate of fluid phase pinocytosis, a significant decrease in the efficiency of chemotactic aggregation, and a decrease in cellular F-actin content. These results identify a complex that links key players in the nucleation and termination of actin filament assembly with a ubiquitous barbed end-directed motor, indicate that the protein responsible for the formation of this complex is physiologically important, and suggest that previously reported myosin I mutant phenotypes in Dictyostelium may be due, at least in part, to defects in the assembly state of actin. We propose that p116 and Acan 125, along with homologues identified in Caenorhabditis elegans, Drosophila, mouse, and man, be named CARMIL proteins, for capping protein, Arp2/3, and myosin I linker. C1 NHLBI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. RP Hammer, JA (reprint author), NHLBI, Cell Biol Lab, NIH, Bldg 3,Room B1-22, Bethesda, MD 20892 USA. NR 77 TC 129 Z9 139 U1 0 U2 1 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD JUN 25 PY 2001 VL 153 IS 7 BP 1479 EP 1497 DI 10.1083/jcb.153.7.1479 PG 19 WC Cell Biology SC Cell Biology GA 447QJ UT WOS:000169583700013 PM 11425877 ER PT J AU Uwaifo, GI Robey, PG Akintoye, SO Collins, MT AF Uwaifo, GI Robey, PG Akintoye, SO Collins, MT TI Clinical picture: Fuel on the fire SO LANCET LA English DT Editorial Material C1 NIH, Dev Endocrinol Branch, Bethesda, MD 20892 USA. NIH, Craniofacial & Skeletal Dis Branch, Bethesda, MD 20892 USA. RP Uwaifo, GI (reprint author), NIH, Dev Endocrinol Branch, Bldg 10, Bethesda, MD 20892 USA. RI Robey, Pamela/H-1429-2011; Uwaifo, Gabriel/M-2361-2016 OI Robey, Pamela/0000-0002-5316-5576; Uwaifo, Gabriel/0000-0002-6962-9304 NR 0 TC 10 Z9 10 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 JUN 23 PY 2001 VL 357 IS 9273 BP 2011 EP 2011 DI 10.1016/S0140-6736(00)05121-7 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 446JB UT WOS:000169509200010 PM 11438133 ER PT J AU Morens, DM Katz, AR Melish, ME AF Morens, DM Katz, AR Melish, ME TI The fourth disease, 1900-1881, RIP SO LANCET LA English DT Letter C1 NIAID, NIH, Bethesda, MD 20815 USA. Univ Hawaii, John A Burns Sch Med, Honolulu, HI 96822 USA. RP Morens, DM (reprint author), NIAID, NIH, Bethesda, MD 20815 USA. NR 5 TC 0 Z9 0 U1 1 U2 1 PU LANCET LTD PI LONDON PA 84 THEOBALDS RD, LONDON WC1X 8RR, ENGLAND SN 0140-6736 J9 LANCET JI Lancet PD JUN 23 PY 2001 VL 357 IS 9273 BP 2059 EP 2059 DI 10.1016/S0140-6736(00)05151-5 PG 1 WC Medicine, General & Internal SC General & Internal Medicine GA 446JB UT WOS:000169509200052 PM 11441870 ER PT J AU Kojima, T Yoshikawa, Y Takada, S Sato, M Nakamura, T Takahashi, N Copeland, NG Gilbert, DJ Jenkins, NA Mori, N AF Kojima, T Yoshikawa, Y Takada, S Sato, M Nakamura, T Takahashi, N Copeland, NG Gilbert, DJ Jenkins, NA Mori, N TI Genomic organization of the Shc-related phosphotyrosine adapters and characterization of the full-length Sck/ShcB: Specific association of p68-Sck/ShcB with pp135 SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE adapter; genome; N-Shc; phosphotyrosine; PTB; Sck; SH2; ShcB; ShcC; signal transduction; Src ID PROTEIN-PROTEIN INTERACTIONS; GENETIC-LINKAGE MAP; SIGNAL-TRANSDUCTION; N-SHC; MOUSE; PATHWAY; BRAIN; CONSERVATION; CHROMOSOME-3; EXPRESSION AB The She gene family is an emerging family, containing at least three members designated Shc/ShcA, Sck/Sli/ShcB, N-Shc/Rai/ShcC in mammals. In this study, we determined the genomic organization of the mouse She family. Coding regions of ShcA, B, and C each comprised 12 exons, spanned approximately 6, 20, and 65 kb, and located on chromosome 3, 10, and 13, respectively. Based on this genome analysis, we determined the full-length structure of mouse Sck/ShcB as a 68-kD protein. We found that the 68-kD full-length Sck/ShcB was more efficiently phosphorylated upon EGF treatment than the previously-analyzed CH2-deleted form. We also found that Sck specifically interacted with a 135-kD phosphoprotein (pp135) through its SH2 domain following membrane depolarization. The Sck-pp135 interaction was reduced by Src kinase inhibitors. These results suggest that Sck, but not N-Shc nor She, transmit signals in conjunction with pp135 following Src activation and/or calcium entry in the cell. (C) 2001 Academic Press. C1 NILS, Dept Mol Genet Res, Morioka, Aichi 4748522, Japan. NAIST, Dept Biosci, Takayama, Nara 6300101, Japan. Kyoto Univ, Fac Sci, Ctr Dev Biol, Kyoto 6068224, Japan. NCI, Frederick Canc Res & Dev Ctr, ABL Basic Res Program, Mammalian Genet Lab, Ft Detrick, MD 21702 USA. RP Mori, N (reprint author), Natl Inst Longev Sci, Dept Mol Genet, Morioka, Aichi 4748522, Japan. NR 33 TC 19 Z9 19 U1 1 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 JUN 22 PY 2001 VL 284 IS 4 BP 1039 EP 1047 DI 10.1006/bbrc.2001.5080 PG 9 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 447JM UT WOS:000169568200027 PM 11409899 ER PT J AU Bogdanov, KY Vinogradova, TM Lakatta, EG AF Bogdanov, KY Vinogradova, TM Lakatta, EG TI Sinoatrial nodal cell ryanodine receptor and Na+-Ca2+ exchanger - Molecular partners in pacemaker regulation SO CIRCULATION RESEARCH LA English DT Article DE sinoatrial node; automaticity; ryanodine receptor; Na+-Ca2+ exchange ID SARCOPLASMIC-RETICULUM; INTRACELLULAR CALCIUM; CA2+ RELEASE; MYOCYTES; MECHANISMS; AUTOMATICITY; FLUCTUATIONS; MUSCLE; HEART AB The rate of spontaneous diastolic depolarization (DD) of sinoatrial nodal cells (SANCs) that triggers recurrent action-potentials (APs) is a fundamental aspect of the heart's pacemaker. Here, in experiments on isolated SANCs. using confocal microscopy combined with a patch clamp technique, we show that ryanodine receptor Ca2+ release during the DD produces a localized subsarcolemmal Ca2+ increase that spreads in a wavelike manner by Ca2+-induced Ca2+ release and produces an inward current via the Na+-Ca2+ exchanger (NCX). Ryanodine, a blocker of the sarcoplasmic reticulum Ca2+ release channel, in a dose-dependent manner reduces the SANG beating rate with an IC50 of 2.6 mu mol/L and abolishes the local Ca2+ transients that precede the AP upstroke. In voltage-clamped cells in which the DD was simulated by voltage ramp, 3 mu mol/L ryanodine decreased an inward current during the voltage ramp by 1.6 +/-0.3 pA/pF (SEMI n = 4) leaving the peak of L-type Ca2+ current unchanged. Likewise, acute blockade of the NCX (via rapid substitution of bath Na+ by Li+) abolished SANG beating and reduced the inward current to a similar extent (1.7 +/-0.4 pA/pF, n = 4), as did ryanodine. Thus, in addition to activation/inactivation of multiple ion channels, Ca2+ activation of the NCX, because of localized sarcoplasmic reticulum Ca2+ release, is a critical element in a chain of molecular interactions that permits the heartbeat to occur and determines its beating rate. C1 NIA, Gerontol Res Ctr, Cardiovasc Sci Lab, NIH, Baltimore, MD 21224 USA. RP Bogdanov, KY (reprint author), NIA, Gerontol Res Ctr, Cardiovasc Sci Lab, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. NR 27 TC 215 Z9 222 U1 2 U2 31 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 JUN 22 PY 2001 VL 88 IS 12 BP 1254 EP 1258 DI 10.1161/hh1201.092095 PG 5 WC Cardiac & Cardiovascular Systems; Hematology; Peripheral Vascular Disease SC Cardiovascular System & Cardiology; Hematology GA 446YK UT WOS:000169541600008 PM 11420301 ER PT J AU Hoffman, MP Engbring, JA Nielsen, PK Vargas, J Steinberg, Z Karmand, AJ Nomizu, M Yamada, Y Kleinman, HK AF Hoffman, MP Engbring, JA Nielsen, PK Vargas, J Steinberg, Z Karmand, AJ Nomizu, M Yamada, Y Kleinman, HK TI Cell type-specific differences in glycosaminoglycans modulate the biological activity of a heparin-binding peptide (RKRLQVQLSIRT) from the G domain of the laminin alpha 1 chain SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID FIBROBLAST-GROWTH-FACTOR; TERMINAL GLOBULAR DOMAIN; SULFATE PROTEOGLYCANS; SYNTHETIC PEPTIDES; CRYSTAL-STRUCTURE; IDENTIFICATION; DYSTROGLYCAN; SITE; SEQUENCES; SURFACE AB AG73 (RKRLQVQLSIRT), a peptide from the G domain of the laminin alpha1 chain, has diverse biological activities with different cell types, The heparan sulfate side chains of syndecan-1 on human salivary gland cells were previously identified as the cell surface ligand for AG73. We used homologous peptides from the other laminin cu-chains (A2G73-A5G73) to determine whether the bioactivity of the AG73 sequence is conserved. Human salivary gland cells and a mouse melanoma cell line (B16F10) both bind to the peptides, but cell attachment was inhibited by glycosaminoglycans, modified heparin, and sized heparin fragments in a cell type-specific manner. In other assays, AG73, but not the homologous peptides, inhibited branching morphogenesis of salivary glands and B16F10 network formation on Matrigel. We identified residues critical for AG73 bioactivity using peptides with amino acid substitutions and truncations. Fewer residues were critical for inhibiting branching morphogenesis (XKXLXVXXXIRT) than those required to inhibit B16F10 network formation on Matrigel (N-terminal XxRLQVQLSIRT). In addition, surface plasmon resonance analysis identified the C-terminal IRT of the sequence to be important for heparin binding. Structure based sequence alignment predicts AG73 in a P-sheet with the N-terminaI K (Lys(2)) and the C-terminal R (Arg(10)) on the surface of the G domain. In conclusion, we have determined that differences in cell surface glycosaminoglycans and differences in the amino acids in AG73 recognized by cells modulate the biological activity of the peptide and provide a mechanism to explain its cell-specific activities. C1 NIDCR, Craniofacial Dev Biol & Regenerat Branch, NIH, Bethesda, MD 20892 USA. Hokkaido Univ, Grad Sch Environm Earth Sci, Sapporo, Hokkaido 0600810, Japan. RP Hoffman, MP (reprint author), NIDCR, Craniofacial Dev Biol & Regenerat Branch, NIH, 30-430,30 Convent Dr,MSC 4370, Bethesda, MD 20892 USA. NR 48 TC 76 Z9 76 U1 2 U2 10 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 JUN 22 PY 2001 VL 276 IS 25 BP 22077 EP 22085 DI 10.1074/jbc.M100774200 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 444RE UT WOS:000169412700014 PM 11304538 ER PT J AU Huang, CS Ding, M Li, JX Leonard, SS Rojanasakul, Y Castranova, V Vallyathan, V Ju, G Shi, XL AF Huang, CS Ding, M Li, JX Leonard, SS Rojanasakul, Y Castranova, V Vallyathan, V Ju, G Shi, XL TI Vanadium-induced nuclear factor of activated T cells activation through hydrogen peroxide SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID NF-KAPPA-B; TRANSCRIPTION FACTOR NFATP; CYCLOSPORINE-A; SIGNAL-TRANSDUCTION; PROTEIN-1 ACTIVATION; AP-1 TRANSACTIVATION; PLASMA-MEMBRANE; CALCINEURIN; FAMILY; EXPRESSION AB The present study investigated the role of reactive oxygen species (ROS) in activation of nuclear factor of activated T cells (NFAT), a pivotal transcription factor responsible for regulation of cytokines, by vanadium in mouse embryo fibroblast PW cells or mouse epidermal Cl 41 cells. Exposure of cells to vanadium led to the transactivation of NFAT in a time- and dose-dependent manner. Scavenging of vanadium-induced H2O2 with Nacety-L-cyteine (a general antioxidant) or catalase (a specific H2O2 inhibitor) or the chelation of vanadate with deferoxamine, resulted in inhibition of NFAT activation. In contrast, an increase in H2O2 generation by the addition of superoxide dismutase or NADPH enhanced vanadium-induced NEAT activation. This vanadate-mediated H2O2 generation was verified by both electron spin resonance and fluorescence staining assay. These results demonstrate that H2O2 plays an important role in vanadium-induced NFAT transactivation in two different cell types. Furthermore, pretreatment of cells with nifedipine, a calcium channel blocker, inhibited vanadium-induced NFAT activation, whereas A23187 and ionomycin, two calcium ionophores, had synergistic effects with vanadium for NFAT induction, Incubation of cells with cyclosporin A (CsA), a pharmacological inhibitor of the phosphatase calcineurin, blocked vanadium-induced NFAT activation. Ah data show that vanadium induces NFAT activation not only through a calcium-dependent and CsA-sensitive pathway but also involved H2O2 generation, suggesting that H2O2 may be involved in activation of calcium-calcineurin pathways for NFAT activation caused by vanadium exposure. C1 NIOSH, Hlth Effects Lab Div, NIH, Morgantown, WV 26505 USA. NYU, Sch Med, Nelson Inst Environm Med, New York, NY 10016 USA. Fourth Mil Med Univ, Inst Neurosci, Xian 710032, Peoples R China. W Virginia Univ, Dept Basic Pharmaceut Sci, Morgantown, WV 26506 USA. RP Shi, XL (reprint author), NIOSH, Hlth Effects Lab Div, NIH, Morgantown, WV 26505 USA. RI Shi, Xianglin/B-8588-2012; OI Rojanasakul, Yon/0000-0002-8839-6462; Huang, Chuanshu/0000-0003-4133-5096 NR 62 TC 61 Z9 63 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 22 PY 2001 VL 276 IS 25 BP 22397 EP 22403 DI 10.1074/jbc.M010828200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 444RE UT WOS:000169412700058 PM 11292823 ER PT J AU Glazner, GW Camandola, S Geiger, JD Mattson, MP AF Glazner, GW Camandola, S Geiger, JD Mattson, MP TI Endoplasmic reticulum D-myo-inositol 1,4,5-trisphosphate-sensitive stores regulate nuclear factor-kappa B binding activity in a calcium-dependent manner SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID NERVE GROWTH-FACTOR; NECROSIS-FACTOR-ALPHA; BACKPROPAGATING ACTION-POTENTIALS; HIPPOCAMPAL SYNAPTIC PLASTICITY; TRANSCRIPTION FACTOR; GENE-EXPRESSION; SIGNAL-TRANSDUCTION; INTRACELLULAR CALCIUM; PHOSPHATE FORMATION; CEREBRAL-ISCHEMIA AB The transcription factor nuclear factor-kappaB (NF-kappaB) plays critical roles in neuronal survival and plasticity and in activation of immune responses. The activation of NF-kappaB has been closely associated with changes in intracellular calcium levels, but the relationship be tween the two remains unclear. Here we report that inhibition of endoplasmic reticulum (ER) D-myo-inositol 1,4,5-trisphosphate (IP3)-gated calcium release caused decreased basal NF-kappaB DNA-binding activity in cultured rat cortical neurons. Activation of NF-kappaB in response to tumor necrosis factor-alpha and glutamate was completely abolished when IP3 receptors were blocked, and NF-kappaB activation in response to depletion of ER calcium by thapsigargin treatment was also decreased by IP3 receptor blockade, We further investigated the relationship between IP3 receptor activation and NF-kappaB activity using a cell-free system. Microsomes enriched in the ER were isolated from adult rat cerebral cortex, resuspended, and treated with agents that induce or inhibit ER calcium release, They were then recentrifuged, and the supernatant was added to cytoplasmic extract isolated from the same source tissue. We found that microsomes released an NF-kappaB stimulating signal in response to activation of IP3 receptors or inhibition of the ER Ca2+-ATPase, but not in response to ryanodine. Studies of intact cells and cell-free preparations indicated that the signal released from the ER was not calcium and was heat- and trypsin sensitive. Our data suggest that activation of IP3 receptors is required for a major component of both constitutive and inducible NF-kappaB binding activity in neurons and that decreasing ER intraluminal calcium levels triggers release of a diffusible NF-kappaB-activating signal from the ER. C1 NIA, Neurosci Lab, Gerontol Res Ctr, NIH, Baltimore, MD 21224 USA. Univ Manitoba, Fac Med, Dept Pharmacol & Therapeut, Winnipeg, MB R3E 0T6, Canada. Johns Hopkins Univ, Sch Med, Dept Neurosci, Baltimore, MD 21205 USA. RP Mattson, MP (reprint author), NIA, Neurosci Lab, Gerontol Res Ctr, NIH, GRC 4F02,5600 Nathan Shock Dr, Baltimore, MD 21224 USA. RI Mattson, Mark/F-6038-2012 FU NINDS NIH HHS [NS39184]; NINR NIH HHS [NRSA467305] NR 82 TC 21 Z9 22 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 JUN 22 PY 2001 VL 276 IS 25 BP 22461 EP 22467 DI 10.1074/jbc.M101315200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 444RE UT WOS:000169412700066 PM 11309390 ER PT J AU Forgues, M Marrogi, AJ Spillare, EA Wu, CG Yang , Q Yoshida, M Wang, XW AF Forgues, M Marrogi, AJ Spillare, EA Wu, CG Yang , Q Yoshida, M Wang, XW TI Interaction of the hepatitis B virus X protein with the Crm1-dependent nuclear export pathway SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID TATA-BINDING PROTEIN; NF-KAPPA-B; HEPATOCELLULAR-CARCINOMA; MESSENGER-RNA; TRANSACTIVATION FUNCTION; SIGNALING PATHWAY; REV PROTEIN; GENE; DNA; ACTIVATION AB The leucine-rich nuclear export signal (NES) is used to shuttle large cellular proteins from the nucleus to the cytoplasm. The nuclear export receptor Crm1 is essential in this process by recognizing the NES motif. Here, we show that the oncogenic hepatitis B virus (HBV)X protein (HBx) contains a functional NES motif. We found that the predominant cytoplasmic localization of HBx is sensitive to the drug leptomycin B (LMB), which specifically inactivates Crm1. Mutations at the two con served leucine residues to alanine at the NES motif (L98A,L100A) resulted in a nuclear redistribution of HBx. A recombinant HBx protein binds to Crm1 in vitro. In addition, ectopic expression of HBx sequesters Crm1 in the cytoplasm. Furthermore, HBx activates NF kappaB by inducing its nuclear translocation in a NES-dependent manner. Abnormal cytoplasmic sequestration of Crm1, accompanied by a nuclear localization of NF kappaB, was also observed in hepatocytes from HBV-positive liver samples with chronic active hepatitis. We suggest that Crm1 may play a role in HBx-mediated liver carcinogenesis. C1 NCI, Human Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. Univ Tokyo, Dept Biotechnol, Tokyo, Japan. RP Wang, XW (reprint author), NCI, Human Carcinogenesis Lab, NIH, 37 Convent Dr,MSC 4255,Bldg 37,Rm 2C07, Bethesda, MD 20892 USA. RI Wang, Xin/B-6162-2009; Yoshida, Minoru/C-8049-2014 NR 52 TC 60 Z9 69 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 22 PY 2001 VL 276 IS 25 BP 22797 EP 22803 DI 10.1074/jbc.M101259200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 444RE UT WOS:000169412700109 PM 11287420 ER PT J AU Morinaga, N Kaihou, Y Vitale, N Moss, J Noda, M AF Morinaga, N Kaihou, Y Vitale, N Moss, J Noda, M TI Involvement of ADP-ribosylation factor 1 in cholera toxin induced morphological changes of Chinese hamster ovary cells SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID GTP-BINDING-PROTEIN; ENDOPLASMIC-RETICULUM; BREFELDIN-A; INTRACELLULAR TRAFFICKING; PHOSPHOLIPASE-D; GOLGI; MEMBRANE; SUBUNIT; TRANSPORT; KDEL AB ADP-ribosylation factor 1 (ARF1) was originally found as a cofactor in CT-catalyzed ADP-ribosylation of Ga, but is now known to participate in vesicle trafficking. We asked whether ARF1 function in vesicular trafficking is necessary for CT induced morphological changes in Chinese hamster ovary (CHO) cells, which result from increased intracellular cAMCP. Brefeldin A treatment of cells suppressed CT action, confirming a requirement for Golgi integrity. Overexpression of a GFP-ARF1 fusion protein did not affect the morphological changes induced by CT, but changes were reduced in cells over expressing guanine nucleotide exchange-defective ARF1(T31N) or GTP hydrolysis-deficient ARF1(Q71L) mutants. In cells expressing these mutants, 8-bromo-cAMP induced changes similar to those seen in cells transfected with ARF1 or vector. Inhibition of CT action was specific for mutants of ARF1 and not reproduced by analogous mutants of ARF5 or ARF6. ARF1(Q71L) was mostly colocalized with beta COP, but ARF5(Q71L) less so. ARF6(Q67L) did not colocalize with beta COP and was partially associated with the plasma membrane. These data are consistent with the conclusion that ARF1 influenced CT action in cells by its specific function in the vesicular transport pathway used by CT to travel from plasma membrane to Golgi to ER. C1 Chiba Univ, Sch Med, Dept Microbiol 2, Chuo Ku, Chiba 2608670, Japan. INSERM U338, Ctr Neurochim, F-67084 Strasbourg, France. NHLBI, Pulm Crit Care Med Branch, NIH, Bethesda, MD 20892 USA. RP Morinaga, N (reprint author), Chiba Univ, Sch Med, Dept Microbiol 2, Chuo Ku, 1-8-1 Inohana, Chiba 2608670, Japan. RI Vitale, nicolas/G-5967-2014 OI Vitale, nicolas/0000-0002-4752-4907 NR 33 TC 16 Z9 16 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 JUN 22 PY 2001 VL 276 IS 25 BP 22838 EP 22843 DI 10.1074/jbc.M101184200 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 444RE UT WOS:000169412700114 PM 11279243 ER PT J AU Lai, WS Blackshear, PJ AF Lai, WS Blackshear, PJ TI Interactions of CCCH zinc finger proteins with mRNA - Tristetraprolin-mediated Au-rich element-dependent mRNA degradation can occur in the absence of a poly(A) tail SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID COLONY-STIMULATING FACTOR; MESSENGER-RNA DECAY; NUCLEOTIDE-SEQUENCE; GENE; DESTABILIZATION; DEADENYLATION; EXPRESSION; STABILITY; BINDING; TIS11 AB The CCCH family of tandem zinc finger proteins has recently been shown to promote the turnover of certain mRNAs containing class II AU-rich elements (AREs). In the case of one member of this family, tristetraprolin (TTP), absence of the protein in knockout mice leads to stabilization of two mRNAs containing AREs of this type, those encoding tumor necrosis factor alpha (TNF alpha) and granulocyte-macrophage colony-stimulating factor. To bean to decipher the mechanism by which these zinc finger proteins stimulate the breakdown of this class of mRNAs, we co-transfected TTP and its related CCCH proteins into 293 cells with vectors encoding full-length TNF alpha, granulocyte-macrophage colony-stimulating factor, and interleukin-3 mRNAs. Go-expression of the CCCH proteins caused the rapid turnover of these ARE-containing mRNAs and also promoted the accumulation of stable breakdown intermediates that were truncated at the 3'-end of the mRNA, even further 5' than the 5'-end of the poly(A) tail. To determine whether an intact poly(A) tail was necessary for TTP to promote this type of mRNA degradation, we inserted the TNF alpha ARE into a nonpolyadenylated histone mRNA and also attached a histone 3'-end-processing sequence to the 3'-end of nonpolyadenylated interleukin-3 and TNF alpha mRNAs. In all three cases, TTP stimulated the turnover of the ARE-containing mRNAs, despite the demonstrated absence of a poly(A) tail. These studies indicate that members of this class of CCCH proteins can promote class II ARE containing mRNA turnover even in the absence of a poly(A) tail, suggesting that the processive removal of the poly(A) tail may not be required for this type of CCCH protein-stimulated mRNA turnover. C1 NIEHS A205, Off Clin REs & Lab Signal Transduct, NIH, Res Triangle Pk, NC 27709 USA. Duke Univ, Med Ctr, Dept Med & Biochem, Durham, NC 27710 USA. RP Blackshear, PJ (reprint author), NIEHS A205, Off Clin REs & Lab Signal Transduct, NIH, 111 Alexandre Dr, Res Triangle Pk, NC 27709 USA. NR 23 TC 104 Z9 106 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 JUN 22 PY 2001 VL 276 IS 25 BP 23144 EP 23154 DI 10.1074/jbc.M100680200 PG 11 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 444RE UT WOS:000169412700155 PM 11279239 ER PT J AU Niranjan, PS Forbes, JG Greer, SC Dudowicz, J Freed, KF Douglas, JF AF Niranjan, PS Forbes, JG Greer, SC Dudowicz, J Freed, KF Douglas, JF TI Thermodynamic regulation of actin polymerization SO JOURNAL OF CHEMICAL PHYSICS LA English DT Article ID LIVING POLYMERIZATION; LATTICE MODEL; PHASE-SEPARATION; BINDING AB A Flory-Huggins-type lattice model of actin polymerization under equilibrium conditions is employed to analyze new spectroscopic measurements for the extent of actin polymerization Phi as a function of temperature T, salt concentration [KCl], and the initial concentration of actin monomers [G(0)]. The theory subsumes existing mechanisms for actin monomer initiation, dimerization, and chain propagation. The extent of polymerization Phi increases with T to an unanticipated maximum, and the calculations explain this unusual effect as arising from a competition between monomer activation, which diminishes upon heating, and propagating chain growth, which increases upon heating. The actin polymerization is described as a rounded phase transition, and the associated polymerization temperature T-p depends strongly, but nearly linearly on [G(0)] and [KCl] over the concentration regimes investigated. Our findings support the suggestion that physicochemical changes can complement regulatory proteins in controlling actin polymerization in living systems. (C) 2001 American Institute of Physics. C1 Univ Maryland, Dept Chem & Biochem, College Pk, MD 20742 USA. Univ Maryland, Dept Chem Engn, College Pk, MD 20742 USA. NIAMS, Phys Biol Lab, NIH, Bethesda, MD 20892 USA. Univ Chicago, James Franck Inst, Chicago, IL 60637 USA. Univ Chicago, Dept Chem, Chicago, IL 60637 USA. Natl Inst Stand & Technol, Div Polymers, Gaithersburg, MD 20899 USA. RP Greer, SC (reprint author), Univ Maryland, Dept Chem & Biochem, College Pk, MD 20742 USA. NR 27 TC 34 Z9 34 U1 0 U2 9 PU AMER INST PHYSICS PI MELVILLE PA 2 HUNTINGTON QUADRANGLE, STE 1NO1, MELVILLE, NY 11747-4501 USA SN 0021-9606 J9 J CHEM PHYS JI J. Chem. Phys. PD JUN 22 PY 2001 VL 114 IS 24 BP 10573 EP 10576 DI 10.1063/1.1380693 PG 4 WC Chemistry, Physical; Physics, Atomic, Molecular & Chemical SC Chemistry; Physics GA 441HL UT WOS:000169224900002 ER PT J AU Hoppin, JA Guzman, JD Tolbert, PE Flagg, EW AF Hoppin, JA Guzman, JD Tolbert, PE Flagg, EW TI Agricultural exposure history among African-American farmers in Georgia SO JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH-PART A LA English DT Article AB Agricultural exposures differ across the United States by region, calendar time period, and agricultural practice, but most of the published literature focuses on white men in the Midwest. A pilot study was conducted to explore the breadth and diversity of farming practices over time among African-American farmers in Georgia whose exposures may differ in important ways. Using a comprehensive life events calendar questionnaire, 17 male African-American farmers aged 36 to 86 yr residing in southeastern Georgia were interviewed regarding their agricultural history in July 1997. Most men ( 15/17) reported working on multiple farms in their lifetime; 3 men worked on 5 different farms during their lifetime. These farmers reported using more chemicals during their lifetime than farmers in the Midwest. Used motor oil was the most frequently reported insecticide applied to animals; this apparently common practice has not been described in the literature and should be better understood since its use may result in dermal exposure to polyaromatic hydrocarbons. Better characterization of regionally specific farming history and individual farming practices will facilitate studies of the health effects of farming. C1 NIEHS, Epidemiol Branch, Res Triangle Pk, NC 27709 USA. Emory Univ, Rollins Sch Publ Hlth, Atlanta, GA 30322 USA. RP Hoppin, JA (reprint author), NIEHS, Epidemiol Branch, MD A3-05,POB 12233, Res Triangle Pk, NC 27709 USA. RI Tolbert, Paige/A-5676-2015 FU NCI NIH HHS [1R21CA69764-01] NR 8 TC 3 Z9 3 U1 1 U2 7 PU TAYLOR & FRANCIS LTD PI LONDON PA 11 NEW FETTER LANE, LONDON EC4P 4EE, ENGLAND SN 1528-7394 J9 J TOXICOL ENV HEAL A JI J. TOXICOL. ENV. HEALTH PT A PD JUN 22 PY 2001 VL 63 IS 4 BP 237 EP 241 DI 10.1080/15287390151143631 PG 5 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 446YZ UT WOS:000169542900001 PM 11437057 ER PT J AU O'Brien, SJ Eizirik, E Murphy, WJ AF O'Brien, SJ Eizirik, E Murphy, WJ TI Genomics - On choosing mammalian genomes for sequencing SO SCIENCE LA English DT Editorial Material ID PLACENTAL MAMMALS; PROJECT C1 NCI, Lab Genom Divers, Frederick, MD 21702 USA. RP O'Brien, SJ (reprint author), NCI, Lab Genom Divers, Frederick, MD 21702 USA. EM obrien@ncifcrf.gov RI Eizirik, Eduardo/K-8034-2012 OI Eizirik, Eduardo/0000-0002-9658-0999 NR 22 TC 44 Z9 47 U1 2 U2 4 PU AMER ASSOC ADVANCEMENT SCIENCE PI WASHINGTON PA 1200 NEW YORK AVE, NW, WASHINGTON, DC 20005 USA SN 0036-8075 J9 SCIENCE JI Science PD JUN 22 PY 2001 VL 292 IS 5525 BP 2264 EP 2266 DI 10.1126/science.1059393 PG 3 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 445KD UT WOS:000169455900037 PM 11423643 ER PT J AU Hecht, AM Horkay, F Geissler, E AF Hecht, AM Horkay, F Geissler, E TI Neutron scattering investigations on a bimodal polymer gel SO JOURNAL OF PHYSICAL CHEMISTRY B LA English DT Article ID THERMODYNAMIC OBSERVATIONS; SWOLLEN GELS; NETWORKS; TEMPERATURE AB This study investigates structural similarities in polyfluorosilicone (PFSi) gels made by different cross-linking procedures, using small-angle neutron scattering (SANS). The scattering properties of gels prepared from long and short precursor chains and of the un-cross-linked polymer solutions are reported. The gel samples were cross-linked by two methods, hydrosilylation, and condensation curing. The scattering response of a bimodal gel made by hydrosilylation is compared with that of a monomodal gel made by condensation curing having nearly identical shear moduli. Significant differences in the microscopic structure are revealed by SANS. In the bimodal gels made by hydrosilylation, the scattering spectrum can be described by two characteristic length scales, one dynamic and one static. The condensation cured sample contains in addition very large structures, possibly due to solid precipitation as a byproduct of the condensation reaction. For this system at least three correlation lengths are thus required to define the scattering curve. C1 Univ Grenoble 1, Spectrometrie Phys Lab, CNRS, UMR5588, F-38402 St Martin Dheres, France. NICHD, Lab Integrat & Med Biophys, NIH, Bethesda, MD 20892 USA. RP Geissler, E (reprint author), Univ Grenoble 1, Spectrometrie Phys Lab, CNRS, UMR5588, BP 87, F-38402 St Martin Dheres, France. NR 27 TC 22 Z9 22 U1 0 U2 6 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 1089-5647 J9 J PHYS CHEM B JI J. Phys. Chem. B PD JUN 21 PY 2001 VL 105 IS 24 BP 5637 EP 5642 DI 10.1021/jp003546g PG 6 WC Chemistry, Physical SC Chemistry GA 443YQ UT WOS:000169371500010 ER PT J AU Le, SY Liu, WM Chen, JH Maizel, JV AF Le, SY Liu, WM Chen, JH Maizel, JV TI Local thermodynamic stability scores are well represented by a non-central student's t distribution SO JOURNAL OF THEORETICAL BIOLOGY LA English DT Article ID RNA SECONDARY STRUCTURE; PROTEIN-CODING REGIONS; REV TRANS-ACTIVATOR; HEPATITIS-C VIRUS; STATISTICAL SIGNIFICANCE; IMMUNODEFICIENCY VIRUS; HELICOBACTER-PYLORI; TARGET SEQUENCE; MESSENGER-RNA; DNA AB Local folding in mRNAs is closely associated with biological functions. rn this study, we reveal the whole distribution of local thermodynamic stability in the complete genome of the poliovirus P3/Leon/37 and the single-stranded RNA sequences that corresponds to the nucleotide sequence of the complete genome sequence (1 667 867 bp) of Helicobacter pylori (H. pylori) strain 26695. Local thermodynamic stability in the RNA sequences is measured by two standard z-scores, significance score and stability score. To estimate the distribution of thermodynamic stability, a model based on the non-central Student's t distribution has been developed. Significant patterns of extremes that are either much more stable or unstable than expected by chance are detected. Our results indicate that the highly stable and statistically more significant folding regions are predominantly in non-coding sequences in the two genome sequences. Moreover, the highly unstable folding regions, on the contrary, are predominantly in the protein coding sequences of H. pylori.. The observed differences across the complete genomic sequences are statistically very significant by a chi (2)-test. These extreme patterns may be useful in searching for target sequences for long-chain antisense RNA and for locating potential RNA functional elements involved in the regulation of gene expression including translation, mRNA localization and metabolism. (C) 2001 Academic Press. C1 NCI, Lab Expt & Computat Biol, Div Basic Sci, NIH, Frederick, MD 21702 USA. Indiana Univ, Dept Comp & Informat Sci, Indianapolis, IN 46202 USA. NCI, Frederick Biomed Supercomp Ctr, SAIC, FCRDC, Frederick, MD 21702 USA. RP Le, SY (reprint author), NCI, Lab Expt & Computat Biol, Div Basic Sci, NIH, Bldg 469,Room 151, Frederick, MD 21702 USA. NR 42 TC 6 Z9 6 U1 0 U2 0 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-5193 J9 J THEOR BIOL JI J. Theor. Biol. PD JUN 21 PY 2001 VL 210 IS 4 BP 411 EP 423 DI 10.1006/jtbi.2001.2318 PG 13 WC Biology; Mathematical & Computational Biology SC Life Sciences & Biomedicine - Other Topics; Mathematical & Computational Biology GA 447PW UT WOS:000169582500003 PM 11403562 ER PT J AU Reichert, TA Sugaya, N Fedson, DS Glezen, WP Simonsen, L Tashiro, M AF Reichert, TA Sugaya, N Fedson, DS Glezen, WP Simonsen, L Tashiro, M TI Vaccinating Japanese schoolchildren against influenza. Reply SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Letter C1 Entropy Ltd, Upper Saddle River, NJ 07458 USA. Entropy Res Inst, Upper Saddle River, NJ 07458 USA. Nippon Kokan Hosp, Kawasaki, Kanagawa 210, Japan. Aventis Pasteur MSD, F-69367 Lyon, France. Baylor Coll Med, Houston, TX 77030 USA. NIAID, Bethesda, MD 20892 USA. Natl Inst Infect Dis, Tokyo 2080011, Japan. RP Reichert, TA (reprint author), Entropy Ltd, Upper Saddle River, NJ 07458 USA. NR 4 TC 2 Z9 2 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 JUN 21 PY 2001 VL 344 IS 25 BP 1947 EP 1948 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 443VJ UT WOS:000169362100015 ER PT J AU Huo, TI Wang, XW Forgues, M Wu, CG Spillare, EA Giannini, C Brechot, C Harris, CC AF Huo, TI Wang, XW Forgues, M Wu, CG Spillare, EA Giannini, C Brechot, C Harris, CC TI Hepatitis B virus X mutants derived from human hepatocellular carcinoma retain the ability to abrogate p53-induced apoptosis SO ONCOGENE LA English DT Article DE HBx; mutation; p53; apoptosis; carcinogenesis ID NF-KAPPA-B; HBX PROTEIN; TRANSCRIPTION FACTOR; CELL-DEATH; GENE-PRODUCT; P53 PROTEIN; SV40-TRANSFORMED CELLS; GENOTOXIC STRESS; TRANSGENIC MICE; TUMOR-ANTIGEN AB Chronic hepatitis B virus (HBV) infection and the integration of its X gene (HBx) are closely associated with the development of hepatocellular carcinoma (HCC), The integrated HBx frequently is truncated or contains point mutations, Previous studies indicated that these HBx mutants have a diminished co-transactivational activity. We have compared the effects of wildtype (wt) HBx and its naturally occurring mutants derived from human HCCs on transcriptional co-transactivation, apoptosis and interactive effects with p53. We demonstrated that overexpression of mutant, but not wt HBx, is defective in transcriptional co-transactivation of the NF-kappaB-driven luciferase reporter. By using a microinjection technique, the HBx mutants were shown to have an attenuated pro-apoptotic activity. This deficiency may be attributed to multiple mutations in the co-transactivation domain of HBx, that leads to decreased stability of the translated product. However, wt or mutant HBx bind to p53 in vitro and retain their ability to block p53-mediated apoptosis in vivo, which has been implicated as its major tumor suppressor function. The abrogation of p53-mediated apoptosis by integrated HBx mutants may provide a selective clonal advantage for preneoplastic or neoplastic hepatocytes acid contribute to hepatocellular carcinogenesis. C1 NCI, Human Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. Taipei Vet Gen Hosp, Taipei, Taiwan. Natl Yang Ming Univ, Sch Med, Taipei 112, Taiwan. Necker Inst, Paris, France. RP Harris, CC (reprint author), NCI, Human Carcinogenesis Lab, NIH, 37 Convent Dr,MSC 4255,Bldg 37,Room 2C05, Bethesda, MD 20892 USA. RI Wang, Xin/B-6162-2009 NR 92 TC 57 Z9 66 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD JUN 21 PY 2001 VL 20 IS 28 BP 3620 EP 3628 DI 10.1038/sj.onc.1204495 PG 9 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 444LD UT WOS:000169400200003 PM 11439325 ER PT J AU Wu, CG Salvay, DM Forgues, M Valerie, K Farnsworth, J Markin, RS Wang, XW AF Wu, CG Salvay, DM Forgues, M Valerie, K Farnsworth, J Markin, RS Wang, XW TI Distinctive gene expression profiles associated with Hepatitis B virus x protein SO ONCOGENE LA English DT Article DE hepatitis B virus; cDNA microarray; liver cancer; chronic active hepatitis ID HUMAN-LIVER-CANCER; HEPATOCELLULAR-CARCINOMA; TRANSGENIC MICE; CDNA MICROARRAY; CELL-DEATH; P53 GENE; HBX GENE; IN-VIVO; HEPATOCARCINOGENESIS; DNA AB Hepatitis B virus (HBV) is a major risk factor for the development of hepatocellular carcinoma (HCC), HBV encodes the potentially oncogenic HBx protein, which mainly functions as a transcriptional co-activator involving in multiple gene deregulations. However, mechanisms underlying HBx-mediated oncogenicity remain unclear, To determine the role(s) of HBx in the early genesis of HCC, we utilized the NCI Oncochip microarray that contains 2208 human cDNA clones to examine the gene expression profiles in either freshly isolated normal primary adult human hepatocytes (Hhep) or an HCC cell line (SK-Hep-l) ecotopically expressing HBx via an adenoviral system. The gene expression profiles also were determined in liver samples from HBV-infected chronic active hepatitis patients when compared with normal liver samples. The microarray results were validated through Northern blot analysis of the expression of selected genes. Using reciprocally labeling hybridizations, scatterplot analysis of gene expression ratios in human primary hepatocytes expressing HBx demonstrates that microarrays are highly reproducible. The comparison of gene expression profiles between HBx-expressing primary hepatocytes and HBV-infected liver samples shows a consistent alteration of many cellular genes including a subset of oncogenes (such as c-myc and c-myb) and tumor suppressor genes (such as APC, p53, WAF1 and WT1), Furthermore, clustering algorithm analysis showed distinctive gene expression profiles in Hhep and SK-Hep-l cells. Our findings are consistent with the hypothesis that the deregulation of cellular genes by oncogenic HBx may be an early event that favors hepatocyte proliferation during liver carcinogenesis. C1 NCI, Human Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. Virginia Commonwealth Univ, Med Coll Virginia, Dept Radiat Oncol, Richmond, VA 23298 USA. Univ Nebraska, Med Ctr, Dept Pathol & Microbiol, Omaha, NE 68198 USA. RP Wang, XW (reprint author), NCI, Human Carcinogenesis Lab, NIH, MSC 4255,Bldg 37,Room 2C25, Bethesda, MD 20892 USA. RI Wang, Xin/B-6162-2009 NR 56 TC 68 Z9 77 U1 1 U2 1 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD JUN 21 PY 2001 VL 20 IS 28 BP 3674 EP 3682 DI 10.1038/sj.onc.1204481 PG 9 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 444LD UT WOS:000169400200008 PM 11439330 ER PT J AU Mills, JL England, L AF Mills, JL England, L TI Food fortification to prevent neural tube defects - Is it working? SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Editorial Material ID FOLIC-ACID; UNITED-STATES; HOMOCYSTEINE; FOLATE; RISK C1 NICHHD, Pediat Epidemiol Sect, Div Epidemiol Stat & Prevent Res, NIH, Bethesda, MD USA. RP Mills, JL (reprint author), 6100 Execut Blvd,Room 7B03, Bethesda, MD 20892 USA. NR 13 TC 14 Z9 14 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 JUN 20 PY 2001 VL 285 IS 23 BP 3022 EP 3023 DI 10.1001/jama.285.23.3022 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 442AR UT WOS:000169263200026 PM 11410102 ER PT J AU Stolzenberg-Solomon, RZ Blaser, MJ Limburg, PJ Perez-Perez, G Taylor, PR Virtamo, J Albanes, D AF Stolzenberg-Solomon, RZ Blaser, MJ Limburg, PJ Perez-Perez, G Taylor, PR Virtamo, J Albanes, D TI Helicobacter pylori seropositivity as a risk factor for pancreatic cancer SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Article ID PAST MEDICAL HISTORY; INFECTION; COHORT; ADENOCARCINOMA; HYBRIDIZATION; ASSOCIATION; HORMONES; STOMACH; PICTURE; GROWTH AB Background: Pancreatic cancer is among the most fatal cancers world-wide and one for which few preventable risk factors have been established. Gastric carriage of Helicobacter pylori, particularly cytotoxin-associated gene-A-positive (CagA+) strains, is known to be a risk factor for peptic ulcer disease and gastric cancer and may have a similar etiologic relationship with pancreatic cancer. Methods: We investigated the association of H. pylori carriage and exocrine pancreatic cancer in a nested case-control study within the Alpha-Tocopherol, Beta-Carotene Cancer Prevention Study cohort of 29133 male Finnish smokers aged 50-69 years at baseline. Case subjects (n = 121) were matched on date of baseline serum collection, study center, age, trial intervention, and completion of the dietary questionnaire to 226 control subjects who were alive at the time the matching case subject was diagnosed and who remained free of cancer, during up to 10 years of follow-up. Levels of immunoglobulin G antibodies to H. pylori whole-cell and CagA+ antigens from stored baseline serum were measured by enzyme-linked immunosorbent assay. Smoking-adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were estimated by use of conditional logistic regression. Statistical tests were two-sided. Results: Seroprevalence of H. pylori was 82% and 73% among case and control subjects, respectively. Compared with seronegative subjects, those with H, pylori or CagA+ strains were at statistically significantly elevated risk of pancreatic cancer (OR = 1.87 [95% CI = 1.05 to 3.34]; OR = 2.01 [95% CI = 1.09 to 3.70], respectively). Conclusions: Our findings support a possible role for H. pylori carriage in the development of exocrine pancreatic cancer. C1 NCI, Nutr Epidemiol Branch, Div Canc Epidemiol & Genet, NIH, Bethesda, MD 20892 USA. NCI, Canc Prevent Studies Branch, Div Clin Sci, NIH, Bethesda, MD 20892 USA. NYU, Sch Med, Dept Med, New York, NY USA. NYU, Sch Med, Dept Microbiol, New York, NY USA. Dept Vet Affairs Med Ctr, New York, NY USA. Natl Publ Hlth Inst, Helsinki, Finland. RP Stolzenberg-Solomon, RZ (reprint author), NCI, Nutr Epidemiol Branch, Div Canc Epidemiol & Genet, NIH, 6120 Execut Blvd,MSC 7026,Rm 7039, Bethesda, MD 20892 USA. RI Albanes, Demetrius/B-9749-2015 FU NCI NIH HHS [N01CN45035, N01CN45165]; NIDDK NIH HHS [R01DK53707] NR 38 TC 92 Z9 97 U1 3 U2 5 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 JUN 20 PY 2001 VL 93 IS 12 BP 937 EP 941 DI 10.1093/jnci/93.12.937 PG 5 WC Oncology SC Oncology GA 442UX UT WOS:000169304300014 PM 11416115 ER PT J AU Solomon, D Schiffman, M AF Solomon, D Schiffman, M TI Re: Comparison of three management strategies for patients with atypical squamous cells of undetermined significance: Baseline results from a randomized trial - Response SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Letter C1 NIH, Rockville, MD 20852 USA. NCI, Breast & Gynecol Canc Res Grp, Div Canc Prevent, Bethesda, MD 20892 USA. NCI, Environm Epidemiol Branch, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. RP Solomon, D (reprint author), NIH, Execut Plaza N,6130 Execut Blvd, Rockville, MD 20852 USA. NR 1 TC 0 Z9 0 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 JUN 20 PY 2001 VL 93 IS 12 BP 951 EP 952 PG 2 WC Oncology SC Oncology GA 442UX UT WOS:000169304300019 ER PT J AU Thiriet, N Jayanthi, S McCoy, M Ladenheim, B Cadet, JL AF Thiriet, N Jayanthi, S McCoy, M Ladenheim, B Cadet, JL TI Methamphetamine increases expression of the apoptotic c-myc and L-myc genes in the mouse brain SO MOLECULAR BRAIN RESEARCH LA English DT Article DE methamphetamine; gene expression; c-myc; L-myc; apoptosis ID DISMUTASE TRANSGENIC MICE; P53-KNOCKOUT MICE; NEUROTOXICITY; CELLS AB To clarify the possible mechanisms by which the recreational drug, methamphetamine (METH), induces apoptosis, we investigated its effects on the expression of Myc apoptotic genes. This paper presents the characterization of c-myc and L-myc gene transcription in the striatum and the cortex. In addition, the expression of the corresponding proteins was also evaluated. Our observations reveal that c-myc and L-myc were up-regulated by METH at both the mRNA and protein levels. Thus, myc transcription factors might be responsible for some aspects of METH-induced apoptotic processes. C1 NIDA, Mol Neuropsychiat Sect, Abuse Intramural Res Program, Baltimore, MD 21224 USA. RP Cadet, JL (reprint author), NIDA, Mol Neuropsychiat Sect, Abuse Intramural Res Program, Baltimore, MD 21224 USA. NR 16 TC 11 Z9 11 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0169-328X J9 MOL BRAIN RES JI Mol. Brain Res. PD JUN 20 PY 2001 VL 90 IS 2 BP 202 EP 204 DI 10.1016/S0169-328X(01)00093-6 PG 3 WC Neurosciences SC Neurosciences & Neurology GA 444YD UT WOS:000169426900012 ER PT J AU Arioglu, E Taylor, SI AF Arioglu, E Taylor, SI TI Treatment of lipodystrophy with troglitazone - In response SO ANNALS OF INTERNAL MEDICINE LA English DT Letter ID INFECTED PATIENTS C1 NIDDKD, Diabet Branch, Bethesda, MD 20892 USA. RP Arioglu, E (reprint author), NIDDKD, Diabet Branch, Bethesda, MD 20892 USA. NR 3 TC 0 Z9 0 U1 1 U2 1 PU AMER COLL PHYSICIANS PI PHILADELPHIA PA INDEPENDENCE MALL WEST 6TH AND RACE ST, PHILADELPHIA, PA 19106-1572 USA SN 0003-4819 J9 ANN INTERN MED JI Ann. Intern. Med. PD JUN 19 PY 2001 VL 134 IS 12 BP 1153 EP 1154 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 442YU UT WOS:000169313200020 ER PT J AU Pal, A Gu, YJ Pan, SS Ji, XH Singh, SV AF Pal, A Gu, YJ Pan, SS Ji, XH Singh, SV TI C-terminal region amino acid substitutions contribute to catalytic differences between murine class alpha glutathione transferases mCSTA1-1 and mCSTA2-2 toward anti-diol epoxide isomers of benzo [c]phenanthrene SO BIOCHEMISTRY LA English DT Article ID S-TRANSFERASE; RAT-LIVER; ANTICARCINOGENIC ORGANOSULFIDES; HTP-II; CONJUGATION; BENZOPYRENE; EFFICIENCY; 7-BETA,8-ALPHA-DIHYDROXY-9-ALPHA,10-ALPHA-OXY-7,8,9,10-TETRAHYDROBENZO(A )PYRENE; BENZOPHENANTHRENE; STEREOISOMERS AB The molecular basis for catalytic differences between structurally closely related murine class alpha glutathione (GSH) transferases mGSTA1-1 and mGSTA2-2 in the GSH conjugation of anti-diol epoxide isomers of benzo[c]phenanthrene (anti-B[c]PDE) was investigated. GSH conjugation of both (-)- and (+)-enantiomers of anti-B[c]PDE was observed in the presence of mGSTA1-1 (60 and 40% GSH conjugation, respectively), whereas mGSTA2-2 exhibited a preference for the (-)-anti-isomer (>97%). In addition, the specific activity of mGSTA2-2 toward the (-)-anti-B[c]PDE isomer was relatively higher than that of mGSTA1-1. The amino acid sequences of mGSTA1-1 and mGSTA2-2 differ at 10 positions that are distributed in three sections. Section I contains amino acid residues in positions 65 and 95; section II contains residues in positions 157, 162, and 169, and section III contains residues in positions 207, 213, 218, 221, and 222. Enzyme activity measurements with chimeras of mGSTA1-1 and mGSTA2-2 revealed that amino acid substitutions in section III account for their differential enantioselectivity and catalytic activity toward anti-B[c]PDE, Site-directed mutagenesis of amino acid residues in section III of mGSTA2-2 with corresponding residues of mGSTA1-1 followed by activity measurements of the wild type and mutated enzymes indicates that leucine 207 and phenylalanine 221 may be critical for the high catalytic activity of mGSTA2-2 toward (-)-anti-B[c]PDE. Molecular modeling studies demonstrated that the active site of mGSTA1-1 accommodates both enantiomers of anti-B[c]PDE, whereas the (-)-anti-isomer interacts more favorably with active site residues in mGSTA2-2, The results of this study clearly indicate that amino acid substitutions in the C-terminal region contribute to catalytic differences between mGSTA1-1 and mGSTA2-2 with respect to anti-B[c]PDE. C1 Univ Pittsburgh, Sch Med, Dept Pharmacol, Pittsburgh, PA 15261 USA. Univ Pittsburgh, Sch Med, Inst Canc, Pittsburgh, PA 15261 USA. NCI, Macromol Crystallog Lab, Frederick, MD 21702 USA. RP Singh, SV (reprint author), Univ Pittsburgh, Sch Med, Dept Pharmacol, S-871 Scaife Hall,Box 130,3550 Terrace St, Pittsburgh, PA 15261 USA. RI Gu, Yijun/B-6017-2012; Ji, Xinhua/C-9664-2012 OI Ji, Xinhua/0000-0001-6942-1514 FU NCI NIH HHS [R01 CA76348, R01 CA55589] NR 35 TC 5 Z9 5 U1 0 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 JUN 19 PY 2001 VL 40 IS 24 BP 7047 EP 7053 DI 10.1021/bi010363r PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 443WU UT WOS:000169366000007 PM 11401548 ER PT J AU Boehm, M Nabel, EG AF Boehm, M Nabel, EG TI Cell cycle and cell migration - New pieces to the puzzle SO CIRCULATION LA English DT Editorial Material DE editorials; cell cycles; cyclins ID PROLIFERATION; RAPAMYCIN; REGULATORS; INHIBITORS; P27(KIP1); GROWTH C1 NHLBI, NIH, Vasc Biol Branch, Bethesda, MD 20892 USA. RP Nabel, EG (reprint author), NHLBI, NIH, Vasc Biol Branch, Bldg 10-8C 103, Bethesda, MD 20892 USA. NR 17 TC 39 Z9 42 U1 0 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 JUN 19 PY 2001 VL 103 IS 24 BP 2879 EP 2881 PG 3 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 445AV UT WOS:000169436000005 PM 11413073 ER PT J AU Hughes, SH AF Hughes, SH TI Molecular matchmaking: NNRTIs can enhance the dimerization of HIV type 1 reverse transcriptase SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Editorial Material ID IMMUNODEFICIENCY-VIRUS TYPE-1; NONNUCLEOSIDE INHIBITORS; ANGSTROM RESOLUTION; CRYSTAL-STRUCTURE; HETERODIMER; MECHANISM; DNA C1 NCI, Frederick, MD 21702 USA. RP Hughes, SH (reprint author), NCI, POB B, Frederick, MD 21702 USA. NR 10 TC 6 Z9 6 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 JUN 19 PY 2001 VL 98 IS 13 BP 6991 EP 6992 DI 10.1073/pnas.141222698 PG 2 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 445KL UT WOS:000169456600004 PM 11416176 ER PT J AU Requena, JR Groth, D Legname, G Stadtman, ER Prusiner, SB Levine, RL AF Requena, JR Groth, D Legname, G Stadtman, ER Prusiner, SB Levine, RL TI Copper-catalyzed oxidation of the recombinant SHa(29-231) prion protein SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID SUPEROXIDE-DISMUTASE; METHIONINE RESIDUES; MASS-SPECTROMETRY; BINDING-SITES; DISEASE; AGGREGATION; CONVERSION; STRESS AB Metal-catalyzed oxidation may result in structural damage to proteins and has been implicated in aging and disease, including neurological disorders such as Alzheimer's disease and amyotrophic lateral sclerosis. The selective modification of specific amino acid residues with high metal ion affinity leads to subtle structural changes that are not easy to detect but may have dramatic consequences on physical and functional properties of the oxidized protein molecules. PrP contains a histidine-rich octarepeat domain that binds copper. Because copper-binding histidine residues are particularly prone to metal-catalyzed oxidation, we investigated the effect of this reaction on the recombinant prion protein SHaPrP(29-231), Using Cu2+/ascorbate, we oxidized SHaPrP(29-231) in vitro. Oxidation was demonstrated by liquid chromatography/mass spectrometry, which showed the appearance of protein species of higher mass, including increases in multiples of 16, characteristic of oxygen incorporation. Digestion studies using Lys C indicate that the 29-101 region, which includes the histidine-containing octarepeats, is particularly affected by oxidation. Oxidation was time- and copper concentration-dependent and was evident with copper concentrations as low as 1 muM. Concomitant with oxidation, SHaPrP(29-231) suffered aggregation and precipitation, which was nearly complete after 15 min, when the prion protein was incubated at 37 degreesC with a 6-fold molar excess of Cu2+. These findings indicate that PrP, a copper-binding protein, may be particularly susceptible to metal-catalyzed oxidation and that oxidation triggers an extensive structural transition leading to aggregation. C1 NHLBI, Biochem Lab, NIH, Bethesda, MD 20892 USA. Univ Calif San Francisco, Inst Neurodegenerat Dis, San Francisco, CA 94143 USA. RP Requena, JR (reprint author), NHLBI, Biochem Lab, NIH, 50 South Dr,Room 2351 MSC 8012, Bethesda, MD 20892 USA. RI Levine, Rodney/D-9885-2011; OI Legname, Giuseppe/0000-0003-0716-4393 NR 32 TC 114 Z9 118 U1 0 U2 4 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUN 19 PY 2001 VL 98 IS 13 BP 7170 EP 7175 DI 10.1073/pnas.121190898 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 445KL UT WOS:000169456600035 PM 11404462 ER PT J AU Kuzmin, PI Zimmerberg, J Chizmadzhev, YA Cohen, FS AF Kuzmin, PI Zimmerberg, J Chizmadzhev, YA Cohen, FS TI A quantitative model for membrane fusion based on low-energy intermediates SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID LIPID BILAYERS; PHASE-TRANSITION; HEMAGGLUTININ; TENSION AB The energetics of a fusion pathway is considered, starting from the contact site where two apposed membranes each locally protrude (as "nipples") toward each other. The equilibrium distance between the tips of the two nipples is determined by a balance of physical forces: repulsion caused by hydration and attraction generated by fusion proteins. The energy to create the initial stalk, caused by bending of cis monolayer leaflets, is much less when the stalk forms between nipples rather than parallel flat membranes. The stalk cannot, however, expand by bending deformations alone, because this would necessitate the creation of a hydrophobic void of prohibitively high energy. But small movements of the lipids out of the plane of their monolayers allow transformation of the stalk into a modified stalk. This intermediate, not previously considered, is a low-energy structure that can reconfigure into a fusion pore via an additional intermediate, the prepore. The lipids of this latter structure are oriented as in a fusion pore, but the bilayer is locally compressed. All membrane rearrangements occur in a discrete local region without creation of an extended hemifusion diaphragm. Importantly, all steps of the proposed pathway are energetically feasible. C1 NICHHD, Lab Cellular & Mol Biophys, NIH, Bethesda, MD 20892 USA. AN Frumkin Electrochem Inst, Moscow 117071, Russia. Rush Med Coll, Dept Physiol & Mol Biophys, Chicago, IL 60612 USA. RP Zimmerberg, J (reprint author), NICHHD, Lab Cellular & Mol Biophys, NIH, Bldg 10,Room 10D14,10 Ctr Dr,MSC 1855, Bethesda, MD 20892 USA. EM jorhz@helix.nih.gov RI Wunder, Stephanie/B-5066-2012; Chizmadzhev, Yuri/L-1984-2013; Zdilla, Michael/B-4145-2011 FU FIC NIH HHS [R03 TW00715]; NIGMS NIH HHS [GM 27367, R01 GM027367] NR 26 TC 209 Z9 216 U1 2 U2 19 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 JUN 19 PY 2001 VL 98 IS 13 BP 7235 EP 7240 DI 10.1073/pnas.121191898 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 445KL UT WOS:000169456600046 PM 11404463 ER PT J AU Liu, ZJ Haleem-Smith, H Chen, HX Metzger, H AF Liu, ZJ Haleem-Smith, H Chen, HX Metzger, H TI Unexpected signals in a system subject to kinetic proofreading SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID HIGH-AFFINITY; IMMUNOGLOBULIN-E; MAST-CELLS; RBL-2H3 CELLS; IGE RECEPTOR; KINASE; MECHANISM; ANTIBODY; RELEASE AB When multivalent ligands attach to IgEs bound tot the! receptors with high affinity for IgE on mast cells, the receptors aggregate. tyrosines on the receptors become phosphorylated, and a variety of cellular responses are stimulated. Prior studies, confirmed here, demonstrated that the efficiency with which later events are generated from earlier ones is inversely related to the dissociation rate of the aggregating ligand. This finding suggests that the cellular responses are constrained by a "kinetic proofreading" regimen. We have now observed an apparent exception to this rule. Doses of the rapidly or slowly dissociating ligands that generated equivalent levels of tyrosine-phosphorylated receptors comparably stimulated a putatively distal event: transcription of the gene for monocyte chemoattractant protein 1. Possible explanations of this apparent anomaly were explored. C1 NIAMSD, Arthrit & Rheumatism Branch, Sect Chem Immunol, NIH, Bethesda, MD 20892 USA. RP Metzger, H (reprint author), NIAMSD, Arthrit & Rheumatism Branch, Sect Chem Immunol, NIH, Bethesda, MD 20892 USA. NR 27 TC 37 Z9 37 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 JUN 19 PY 2001 VL 98 IS 13 BP 7289 EP 7294 DI 10.1073/pnas.121171998 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 445KL UT WOS:000169456600055 PM 11371625 ER PT J AU Hlavacek, WS Redondo, A Metzger, H Wofsy, C Goldstein, B AF Hlavacek, WS Redondo, A Metzger, H Wofsy, C Goldstein, B TI Kinetic proofreading models for cell signaling predict ways to escape kinetic proofreading SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID HIGH-AFFINITY RECEPTOR; FC-EPSILON-RI; IGE RECEPTOR; MULTIVALENT ANTIGEN; TRANSDUCTION; AGGREGATION; MECHANISM; KINASES; PHOSPHORYLATION; DISSOCIATION AB In the context of cell signaling, kinetic proofreading was introduced to explain how cells can discriminate among ligands based on a kinetic parameter, the ligand-receptor dissociation rate constant. In the kinetic proofreading model of cell signaling, responses occur only when a bound receptor undergoes a complete series of modifications. If the ligand dissociates prematurely, the receptor returns to its basal state and signaling is frustrated. Mle extend the model to deal with systems where aggregation of receptors is essential to signal transduction, and present a version of the model for systems where signaling depends on an extrinsic kinase. We also investigate the kinetics of signaling molecules, "messengers." that are generated by aggregated receptors but do not remain associated with the receptor complex. We show that the extended model predicts modes of signaling that exhibit kinetic discriimination for some range of parameters but for other parameter values show little or no discrimination and thus escape kinetic proofreading. We compare model predictions with experimental data. C1 Univ Calif Los Alamos Natl Lab, Div Theoret, Theoret Biol & Biophys Grp, Los Alamos, NM 87545 USA. Univ Calif Los Alamos Natl Lab, Div Theoret, Theoret Chem Grp, Los Alamos, NM 87545 USA. NIAMSD, Arthrit & Rheumatism Branch, NIH, Bethesda, MD 20892 USA. Univ New Mexico, Dept Math & Stat, Albuquerque, NM 87131 USA. RP Goldstein, B (reprint author), Univ Calif Los Alamos Natl Lab, Div Theoret, Theoret Biol & Biophys Grp, POB 1663, Los Alamos, NM 87545 USA. FU NIGMS NIH HHS [R01 GM035556, R37 GM035556, GM35556] NR 28 TC 39 Z9 40 U1 0 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 JUN 19 PY 2001 VL 98 IS 13 BP 7295 EP 7300 DI 10.1073/pnas.121172298 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 445KL UT WOS:000169456600056 PM 11390967 ER PT J AU Kostrouchova, M Krause, M Kostrouch, Z Rall, JE AF Kostrouchova, M Krause, M Kostrouch, Z Rall, JE TI Nuclear hormone receptor CHR3 is a critical regulator of all four larval molts of the nematode Caenorhabditis elegans SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE molting; development ID COLLAGEN GENE DPY-7; DOUBLE-STRANDED-RNA; DROSOPHILA METAMORPHOSIS; GENOME SEQUENCE; ORPHAN RECEPTOR; C-ELEGANS; DHR3; EXPRESSION; SUPERFAMILY; EMBRYOGENESIS AB CHR3 (nhr-23, NF1F4), the homologue of Drosophila DHR3 and mammalian ROR/RZR/RevErbA nuclear hormone receptors. is important for proper epidermal development and molting in the nematode Caenorhabditis elegans. Disruption of CHR3 (nhr-23) function leads to developmental changes, including incomplete molting and a short, fat (dumpy) phenotype. Here, we studied the role of CHR3 during larval development by using expression assays and RNA-mediated interference. We show that the levels of expression of CHR3 (nhr-23) cycle during larval development and reduction of CHR3 function during each intermolt period result in defects at all subsequent molts. Assaying candidate gene expression in populations of animals treated with CHR3 (nhr-23) RNA-mediated interference has identified dpy-7 as a potential gene acting downstream of CHR3. These results define CHR3 as a critical regulator of all C. elegans molts and begin to define the molecular pathway for its function. C1 NIDDKD, Diabet Branch, NIH, Bethesda, MD 20892 USA. NIDDKD, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. Charles Univ, Fac Med 1, Inst Inherited Metab Disorders, Lab Mol Pathol, CR-11636 Prague, Czech Republic. Charles Univ, Fac Med 1, Lab Mol Biol & Genet, CR-11636 Prague, Czech Republic. RP Rall, JE (reprint author), NIDDKD, Diabet Branch, NIH, Bldg 10,Room 95213,9000 Rockville Pike, Bethesda, MD 20892 USA. OI Krause, Michael/0000-0001-6127-3940 NR 50 TC 79 Z9 81 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 JUN 19 PY 2001 VL 98 IS 13 BP 7360 EP 7365 DI 10.1073/pnas.131171898 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 445KL UT WOS:000169456600067 PM 11416209 ER PT J AU Cao, B Su, YL Oskarsson, M Zhao, P Kort, EJ Fisher, RJ Wang, LM Vande Woude, GF AF Cao, B Su, YL Oskarsson, M Zhao, P Kort, EJ Fisher, RJ Wang, LM Vande Woude, GF TI Neutralizing monoclonal antibodies to hepatocyte growth factor/scatter factor (HGF/SF) display antitumor activity in animal models SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID FACTOR SCATTER FACTOR; C-MET RECEPTOR; PROTOONCOGENE PRODUCT; HUMAN GLIOBLASTOMA; TYROSINE KINASE; FACTOR PROTECTS; HUMAN GLIOMAS; ANGIOGENESIS; EXPRESSION; METASTASIS AB The hepatocyte growth factor (HGF/SF) receptor, Met. regulates mitogenesis, motility. and morphogenesis in a cell type-dependent fashion. Activation of Met via autocrine, paracrine, or mutational mechanisms can lead to tumorigenesis and metastasis and numerous studies have linked inappropriate expression of this ligand-receptor pair to most types of human solid tumors. To prepare mAbs to human HGF/SF. mice were immunized with native and denatured preparations of the ligand. Recloned mAbs were tested in vitro for blocking activity against scattering and branching morphogenesis. Our results show that no single mAb was capable of neutralizing the in vitro activity of HGF/SF, and that the ligand possesses a minimum of three epitopes that must be blocked to prevent Met tyrosine kinase activation. In vivo, the neutralizing mAb combination inhibited s.c. growth in athymic nu/nu mice of tumors dependent on an autocrine Met-HGF/SF loop. Importantly, growth of human glioblastoma multiforme xenografts expressing Met and HGF/SF were markedly reduced in the presence of HGF/ SF-neutralizing mAbs. These results suggest interrupting autocrine and/or paracrine Met-HGF/SF signaling in tumors dependent on this pathway is a possible intervention strategy. C1 Van Andel Res Inst, Grand Rapids, MI 49503 USA. NCI, Frederick Canc Res & Dev Ctr, Ft Detrick, MD 21702 USA. RP Vande Woude, GF (reprint author), Van Andel Res Inst, 333 Bostwick NE, Grand Rapids, MI 49503 USA. NR 50 TC 173 Z9 181 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 JUN 19 PY 2001 VL 98 IS 13 BP 7443 EP 7448 DI 10.1073/pnas.131200498 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 445KL UT WOS:000169456600082 PM 11416216 ER PT J AU Czar, MJ Kersh, EN Mijares, LA Lanier, G Lewis, J Yap, G Chen, A Sher, A Duckett, CS Ahmed, R Schwartzberg, PL AF Czar, MJ Kersh, EN Mijares, LA Lanier, G Lewis, J Yap, G Chen, A Sher, A Duckett, CS Ahmed, R Schwartzberg, PL TI Altered lymphocyte responses and cytokine production in mice deficient in the X-linked lymphoproliferative disease gene SH2D1A/DSHP/SAP SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID ACTIVATION MOLECULE SLAM; BARR-VIRUS INFECTION; NATURAL-KILLER-CELLS; T-CELLS; GAMMA PRODUCTION; VIRAL-INFECTION; ENCODING GENE; CUTTING EDGE; SH2D1A GENE; 2B4 AB We have introduced a targeted mutation in SH2D1A/DSHP/SAP, the gene responsible for the human genetic disorder X-linked lymphoproliferative disease (XLP). SLAM-associated protein (SAP)-deficient mice had normal lymphocyte development, but on challenge with infectious agents, recapitulated features of XLP. Infection of SAP(-) mice with lymphocyte choriomeningitis virus (LCMV) or Toxoplasma gondii was associated with increased T cell activation and IFN-gamma production, as well as a reduction of Ig-secreting cells. Anti-CD3-stimulated splenocytes from uninfected SAP(-) mice produced increased IFN-gamma and decreased IL-4, findings supported by decreased serum IgE levels in vivo. The Th1 skewing of these animals suggests that cytokine misregulation may contribute to phenotypes associated with mutation of SH2D1A/SAP. C1 NHGRI, NIH, Bethesda, MD 20892 USA. NCI, NIH, Bethesda, MD 20892 USA. NIAID, NIH, Bethesda, MD 20892 USA. Emory Univ, Dept Microbiol & Immunol, Atlanta, GA 30322 USA. Emory Univ, Emory Vaccine Ctr, Atlanta, GA 30322 USA. RP Schwartzberg, PL (reprint author), NHGRI, NIH, 49-4A38, Bethesda, MD 20892 USA. NR 34 TC 174 Z9 179 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 JUN 19 PY 2001 VL 98 IS 13 BP 7449 EP 7454 DI 10.1073/pnas.131193098 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 445KL UT WOS:000169456600083 PM 11404475 ER PT J AU Dreijerink, K Braga, E Kuzmin, I Geil, L Duh, FM Angeloni, D Zbar, B Lerman, MI Stanbridge, EJ Minna, JD Protopopov, A Li, JF Kashuba, V Klein, G Zabarovsky, ER AF Dreijerink, K Braga, E Kuzmin, I Geil, L Duh, FM Angeloni, D Zbar, B Lerman, MI Stanbridge, EJ Minna, JD Protopopov, A Li, JF Kashuba, V Klein, G Zabarovsky, ER TI The candidate tumor suppressor gene, RASSF1A, from human chromosome 3p21.3 is involved in kidney tumorigenesis SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID RENAL-CELL CARCINOMA; DOUBLE-LOSS MECHANISM; SHORT ARM; DNA METHYLATION; LINDAU DISEASE; VHL; CANCER; REGION; HETEROZYGOSITY; LOCUS AB Clear cell-type renal cell carcinomas (clear RCC) are characterized almost universally by loss of heterozygosity on chromosome 30, which usually involves any combination of three regions: 3p25-p26 (harboring the VHL gene), 3p12-p14.2 (containing the FHIT gene), and 3p21-p22, implying inactivation of the resident tumor-suppressor genes (TSGs). For the 3p21-p22 region, the affected TSGs remain, at present, unknown. Recently, the RAS association family 1 gene (isoform RASSF1A), located at 3p21.3, has been identified as a candidate lung and breast TSC. In this report, we demonstrate aberrant silencing by hypermethylation of RASSF1A in both VHL-caused clear RCC tumors and clear RCC without VHL inactivation. We found hypermethylation of RASSF1A's CC-rich putative promoter region in most of analyzed samples, including 39 of 43 primary tumors (91%). The promoter was methylated partially or completely in all 18 RCC cell lines analyzed. Methylation of the CC-rich putative RASSF1A promoter region and loss of transcription of the corresponding mRNA were related causally. RASSF1A expression was reactivated after treatment with 5-aza-2 ' -deoxycytidine. Forced expression of RASSF1A transcripts in KRC/Y, a renal carcinoma cell line containing a normal and expressed VHL gene, suppressed growth on plastic dishes and anchorage-independent colony formation in soft agar. Mutant RASSF1A had reduced growth suppression activity significantly. These data suggest that RASSF1A is the candidate renal TSG gene for the 3p21.3 region. C1 NCI, Intramural Res Support Program, Sci Applicat Int Corp, Frederick, MD 21702 USA. NCI, Immunobiol Lab, Sci Applicat Int Corp, Frederick, MD 21702 USA. Karolinska Inst, Microbiol & Tumor Biol Ctr, S-17177 Stockholm, Sweden. Univ Calif Irvine, Dept Microbiol & Mol Genet, Irvine, CA 92697 USA. Univ Texas, SW Med Ctr, Hamon Ctr Therapeut Oncol Res, Dallas, TX 75390 USA. Russian Acad Sci, VA Engelhardt Mol Biol Inst, Moscow 117984, Russia. Karolinska Inst, Ctr Genom Res, S-17177 Stockholm, Sweden. RP Kuzmin, I (reprint author), NCI, Intramural Res Support Program, Sci Applicat Int Corp, Bldg 560,Room 12-71, Frederick, MD 21702 USA. RI Zabarovsky, Eugene/A-6645-2010; Braga, Eleonora/P-5574-2016 FU NCI NIH HHS [P50 CA070907, R01 CA071618, CA71618, N01-CO-56000, P50 CA70907, CA19104] NR 29 TC 233 Z9 250 U1 0 U2 5 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 JUN 19 PY 2001 VL 98 IS 13 BP 7504 EP 7509 DI 10.1073/pnas.131216298 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 445KL UT WOS:000169456600092 PM 11390984 ER PT J AU Reid, SD Green, NM Buss, JK Lei, B Musser, JM AF Reid, SD Green, NM Buss, JK Lei, B Musser, JM TI Multilocus analysis of extracellular putative virulence proteins made by group A Streptococcus: Population genetics, human serologic response, and gene transcription SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID GROUP-A STREPTOCOCCUS; FIBRONECTIN-BINDING PROTEIN; SHOCK-LIKE SYNDROME; NUCLEOTIDE SUBSTITUTIONS; LISTERIA-MONOCYTOGENES; NEGATIVE REGULATOR; ESCHERICHIA-COLI; PYOGENES; IDENTIFICATION; EXPRESSION AB Species of pathogenic microbes are composed of an array of evolutionarily distinct chromosomal genotypes characterized by diversity in gene content and sequence (allelic variation). The occurrence of substantial genetic diversity has hindered progress in developing a comprehensive understanding of the molecular basis of virulence and new therapeutics such as vaccines. To provide new information that bears on these issues, 11 genes encoding extracellular proteins in the human bacterial pathogen group A Streptococcus identified by analysis of four genomes were studied. Eight of the 11 genes encode proteins with a LPXTC(L) motif that covalently links Cram-positive virulence factors to the bacterial cell surface. Sequence analysis of the 11 genes in 37 geographically and phylogenetically diverse group A Streptococcos strains cultured from patients with different infection types found that recent horizontal gene transfer has contributed substantially to chromosomal diversity. Regions of the inferred proteins likely to interact with the host were identified by molecular population genetic analysis, and Western immunoblot analysis with sera from infected patients confirmed that they were antigenic. Real-time reverse transcriptase-PCR (TaqMan) assays found that transcription of six of the 11 genes was substantially up-regulated in the stationary phase. In addition, transcription of many genes was influenced by the covR and mga trans-acting gene regulatory loci. Multilocus investigation of putative virulence genes by the integrated approach described herein provides an important strategy to aid microbial pathogenesis research and rapidly identify new targets for therapeutics research. C1 NIAID, Lab Human Bacterial Pathogenesis, Rocky Mt Labs, NIH, Hamilton, MT 59840 USA. RP Musser, JM (reprint author), NIAID, Lab Human Bacterial Pathogenesis, Rocky Mt Labs, NIH, 903 S 4th St, Hamilton, MT 59840 USA. NR 58 TC 36 Z9 36 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 JUN 19 PY 2001 VL 98 IS 13 BP 7552 EP 7557 DI 10.1073/pnas.121188598 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 445KL UT WOS:000169456600100 PM 11416223 ER PT J AU Hwang, CK D'Souza, UM Eisch, AJ Yajima, S Lammers, CH Yang, Y Lee, SH Kim, YM Nestler, EJ Mouradian, MM AF Hwang, CK D'Souza, UM Eisch, AJ Yajima, S Lammers, CH Yang, Y Lee, SH Kim, YM Nestler, EJ Mouradian, MM TI Dopamine receptor regulating factor, DRRF: A zinc finger transcription factor SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID MESSENGER-RNA EXPRESSION; BOX-BINDING-PROTEINS; REPRESSION DOMAINS; CRYSTAL-STRUCTURE; MULTIGENE FAMILY; DNA RECOGNITION; GENE-EXPRESSION; DUAL-FUNCTION; ACTIVATION; SP1 AB Dopamine receptor genes are under complex transcription control, determining their unique regional distribution in the brain. We describe here a zinc finger type transcription factor, designated dopamine receptor regulating factor (DRRF), which binds to Gc and GT boxes in the D-1A and D-2 dopamine receptor promoters and effectively displaces Spl and sp3 from these sequences. Consequently, DRRF can modulate the activity of these dopamine receptor promoters. Highest DRRF mRNA levels are found in brain with a specific regional distribution including olfactory bulb and tubercle, nucleus accumbens, striatum, hippocampus. amygdala, and frontal cortex. Many of these brain regions also express abundant levels of various dopamine receptors. In vivo, DRRF itself can be regulated by manipulations of dopaminergic transmission. Mice treated with drugs that increase extracellular striatal dopamine levels (cocaine), block dopamine receptors (haloperidol), or destroy dopamine terminals (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) show significant alterations in DRRF mRNA. The latter observations provide a basis for dopamine receptor regulation after these manipulations. We conclude that DRRF is important for modulating dopaminergic transmission in the brain. C1 NINDS, Genet Pharmacol Unit, Expt Therapeut Branch, NIH, Bethesda, MD 20892 USA. Univ Texas, SW Med Ctr, Dept Psychiat, Dallas, TX 75390 USA. RP Mouradian, MM (reprint author), NINDS, Genet Pharmacol Unit, Expt Therapeut Branch, NIH, Bethesda, MD 20892 USA. OI Mouradian, M. Maral/0000-0002-9937-412X NR 46 TC 47 Z9 50 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 JUN 19 PY 2001 VL 98 IS 13 BP 7558 EP 7563 DI 10.1073/pnas.121635798 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 445KL UT WOS:000169456600101 PM 11390978 ER PT J AU Nakao, Y Itoh, Y Kuang, TY Cook, M Jehle, J Sokoloff, L AF Nakao, Y Itoh, Y Kuang, TY Cook, M Jehle, J Sokoloff, L TI Effects of anesthesia on functional activation of cerebral blood flow and metabolism SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article DE whisker-to-barrel cortex pathway; cerebral glucose utilization; deoxy[C-14]glucose; functional brain imaging; iodo[C-14]antipyrine ID NITRIC-OXIDE SYNTHASE; GLUCOSE-UTILIZATION; SOMATOSENSORY STIMULATION; NEURONAL MESSENGER; RAT; INHIBITION; BRAIN; CHLORALOSE; GLUTAMATE; CORTEX AB Functional brain mapping based on changes in local cerebral blood flow (ICBF) or glucose utilization (ICMRglc) induced by functional activation is generally carried out in animals under anesthesia, usually alpha -chloralose because of its lesser effects on cardiovascular, respiratory, and reflex functions. Results of studies on the role of nitric oxide (NO) in the mechanism of functional activation of ICBF have differed in unanesthetized and anesthetized animals. NO synthase inhibition markedly attenuates or eliminates the ICBF responses in anesthetized animals but not in unanesthetized animals. The present study examines in conscious rats and rats anesthetized with cy-chloralose the effects of vibrissal stimulation on ICMRglc and ICBF in the whisker-to-barrel cortex pathway and on the effects of NO synthase inhibition with N-G-nitro-L-arginine methyl ester (L-NAME) on the magnitude of the responses. Anesthesia markedly reduced the ICBF and ICMRglc responses in the ventral posteromedial thalamic nucleus and barrel cortex but not in the spinal and principal trigeminal nuclei. L-NAME did not alter the ICBF responses in any of the structures of the pathway in the unanesthetized rats and also not in the trigeminal nuclei of the anesthetized rats. In the thalamus and sensory cortex of the anesthetized rats, where the ICBF responses to stimulation had already been drastically diminished by the anesthesia, L-NAME treatment resulted in loss of statistically significant activation of ICBF by vibrissal stimulation. These results indicate that NO does not mediate functional activation of ICBF under physiological conditions. C1 NIMH, Cerebral Metab Lab, NIH, Bethesda, MD 20892 USA. RP Sokoloff, L (reprint author), NIMH, Cerebral Metab Lab, NIH, Bldg 36,Room 1A-07,36 Convent Dr MSC 4030, Bethesda, MD 20892 USA. NR 43 TC 121 Z9 122 U1 0 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 JUN 19 PY 2001 VL 98 IS 13 BP 7593 EP 7598 DI 10.1073/pnas.121179898 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 445KL UT WOS:000169456600107 PM 11390971 ER PT J AU Shaw, KTY Utsuki, T Rogers, J Yu, QS Sambamurti, K Brossi, A Ge, YW Lahiri, DK Greig, NH AF Shaw, KTY Utsuki, T Rogers, J Yu, QS Sambamurti, K Brossi, A Ge, YW Lahiri, DK Greig, NH TI Phenserine regulates translation of beta-amyloid precursor protein mRNA by a putative interleukin-1 responsive element, a target for drug development SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID SYNTHASE MESSENGER-RNA; ALZHEIMERS-DISEASE; KINASE-C; PHORBOL ESTER; MITOCHONDRIAL ACONITASE; 5'-UNTRANSLATED REGION; NEUROBLASTOMA-CELLS; SECRETASE CLEAVAGE; RETINOIC ACID; HUMAN BRAIN AB The reduction in levels of the potentially toxic amyloid-beta peptide (A beta) has emerged as one of the most important therapeutic goals in Alzheimer's disease. Key targets for this goal are factors that affect the expression and processing of the A beta precursor protein (beta APP). Earlier reports from our laboratory have shown that a novel cholinesterase inhibitor, phenserine, reduces beta APP levels in vivo. Herein, we studied the mechanism of phenserine's actions to define the regulatory elements in beta APP processing. Phenserine treatment resulted in decreased secretion of soluble beta APP and A beta into the conditioned media of human neuroblastoma cells without cellular toxicity. The regulation of beta APP protein expression by phenserine was posttranscriptional as it suppressed beta APP protein expression without altering beta APP mRNA levels. However, phenserine's action was neither mediated through classical receptor signaling pathways, involving extracellular signal-regulated kinase or phosphatidylinositol 3-kinase activation, nor was it associated with the anticholinesterase activity of the drug. Furthermore, phenserine reduced expression of a chloramphenicol acetyltransferase reporter fused to the 5 ' -mRNA leader sequence of beta APP without altering expression of a control chloramphenicol acetyltransferase reporter. These studies suggest that phenserine reduces A beta levels by regulating beta APP translation via the recently described iron regulatory element in the 5 ' -untranslated region of beta APP mRNA, which has been shown previously to be up-regulated in the presence of interleukin-1. This study identifies an approach for the regulation of beta APP expression that can result in a substantial reduction in the level of A beta. C1 NIA, Drug Design & Dev, Neurosci Lab, Baltimore, MD 21224 USA. Harvard Univ, Sch Med, Genet Aging Unit, Boston, MA 02115 USA. Mayo Clin, Dept Pharmacol, Jacksonville, FL 32224 USA. Indiana Univ, Sch Med, Inst Psychiat Res, Dept Psychiat, Indianapolis, IN 46202 USA. RP Greig, NH (reprint author), NIA, Drug Design & Dev, Neurosci Lab, Baltimore, MD 21224 USA. NR 48 TC 149 Z9 153 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 JUN 19 PY 2001 VL 98 IS 13 BP 7605 EP 7610 DI 10.1073/pnas.131152998 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 445KL UT WOS:000169456600109 PM 11404470 ER PT J AU Saul, A Miller, LH AF Saul, A Miller, LH TI A robust neutralization test for Plasmodium falciparum malaria SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Editorial Material ID MEROZOITE SURFACE PROTEIN-1; INHIBIT PARASITE GROWTH; HUMAN-ANTIBODIES; VACCINE ANTIGEN; IN-VITRO; INVITRO; MSP-1 C1 NIAID, MVDU, LPD, NIH, Rockville, MD 20852 USA. RP Saul, A (reprint author), NIAID, MVDU, LPD, NIH, Twinbrook 1 Rm 1113,5640 Fisher Lane, Rockville, MD 20852 USA. RI Saul, Allan/I-6968-2013 OI Saul, Allan/0000-0003-0665-4091 NR 24 TC 4 Z9 4 U1 0 U2 0 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 JUN 18 PY 2001 VL 193 IS 12 BP F51 EP F54 DI 10.1084/jem.193.12.F51 PG 4 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 445TW UT WOS:000169475500002 PM 11413200 ER PT J AU Stocking, EM Sanz-Cervera, JF Unkefer, CJ Williams, RM AF Stocking, EM Sanz-Cervera, JF Unkefer, CJ Williams, RM TI Studies on the biosynthesis of paraherquamide. Construction of the amino acid framework SO TETRAHEDRON LA English DT Article DE biosynthesis; secondary metabolism; feeding experiments ID DIELS-ALDER CYCLOADDITION; VM55599 RING-SYSTEMS; ANTHELMINTIC ACTIVITY; L-PROLINE; PENICILLIUM-CHARLESII; ANTIPARASITIC AGENTS; PRENYL TRANSFERASES; BETA-METHYLPROLINE; BREVIANAMIDE-B; SCLEROTAMIDE AB It has been previously established in this laboratory that the beta -methyl-beta -hydroxyproline moiety of the potent anthelmintic agent paraherquamide A, is biosynthetically derived from L-isoleucine. The downstream events from L-Ile to paraherquamide A have now been investigated. The synthesis of [1-(13)C]-labeled L-beta -methylproline is described by means of a Hoffman-Loeffler-Freytag reaction sequence from [1-(13)C]-L-Ile. This amino acid is shown to be a direct biosynthetic precursor to paraherquamide A by feeding and incorporation experiments in growing cultures of Penicillum fellutanum. Three tryptophan-containing dipeptides of L-beta -methylproline have been constructed: [(13)C(2)]-2-(l,1-dimethyl-2-propenyl)-L-tryptophanyl-3(S)-methyl-L-proline [(13)C(2)]-3(S)-methyl-L-prolyl-2-(l,1-dimethyl-2-propenyl)-L-tryptophan and [(13)C(2)]-cyclo-2-(l,1-dimethyl-2-propenyl)-L-tryptophan-3(S)-methyl-L-proline. [alpha-(15)N, 1-(13)C]-2-(1,1-Dimethyl-2-propenyl)-L-tryptophan was also prepared but none of these substances were found to serve as biosynthetic precursors to paraherquamide A. (C) 2001 Elsevier Science Ltd. All rights reserved. C1 Colorado State Univ, Dept Chem, Ft Collins, CO 80523 USA. Univ Valencia, Dept Quim Inorgan, E-46100 Valencia, Spain. Los Alamos Natl Lab, NIH, Stable Isotope Resource, Los Alamos, NM 87545 USA. RP Williams, RM (reprint author), Colorado State Univ, Dept Chem, Ft Collins, CO 80523 USA. EM rmw@chem.colostate.edu RI Sanz-Cervera, Juan/K-1095-2014 NR 34 TC 23 Z9 24 U1 0 U2 7 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0040-4020 J9 TETRAHEDRON JI Tetrahedron PD JUN 18 PY 2001 VL 57 IS 25 BP 5303 EP 5320 DI 10.1016/S0040-4020(01)00449-5 PG 18 WC Chemistry, Organic SC Chemistry GA 445DP UT WOS:000169443100002 ER PT J AU Cooper, GS Baird, DD Darden, FR AF Cooper, GS Baird, DD Darden, FR TI Measures of menopausal status in relation to demographic, reproductive, and behavioral characteristics in a population-based study of women aged 35-49 years SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Article DE alcohol drinking; menarche; menopause; obesity; parity ID STIMULATING-HORMONE LEVELS; NATURAL MENOPAUSE; PASSIVE SMOKING; HYSTERECTOMY; DETERMINANTS; FSH; MORTALITY; RISK; LIFE; LH AB The purpose of this cross-sectional analysis of women aged 35-49 years from the Third National Health and Nutrition Examination Survey, conducted between 1988 and 1994, was to assess associations with menopausal status based either on menstrual cycle patterns or on elevated (> 20 IU/liter) follicle-stimulating hormone. Menstrual cycle-based menopausal status was defined for women who had not had surgical menopause by months since the last period (<2, 2-12, and > 12 months for pre-, peri-, and postmenopause, respectively). Logistic regression was adjusted for age, smoking, and unilateral oophorectomy, Higher body mass index (greater than or equal to 30 kg/m(2) compared with < 25.0 kg/m(2)) was associated with a lower likelihood of elevated follicle-stimulating hormone (odds ratio (OR) = 0.6, 95% confidence interval (CI): 0.4, 0.9) but this association was not seen with the menstrual measure of menopause. Exercise (three or more times per week) was associated with a lower likelihood of being postmenopausal on the basis of menstrual (OR = 0.3, 95% CI: 0.2, 0.7) and hormonal (OR = 0.6, 95% CI, 0.4, 1.0) measures. Alcohol use also tended to be associated with postmenopausal status by either measure, but not significantly so. There was little evidence of associations with ethnicity, education, age at menarche, number of livebirths, and oral contraceptive use. Menstrual-based definitions of menopause can be misclassified for women with menstrual irregularity. This might explain why obese women were classified menstrually as menopausal while remaining hormonally premenopausal. C1 NIEHS, Epidemiol Branch A305, Res Triangle Pk, NC 27709 USA. Westat Inc, Res Triangle Pk, NC USA. RP Cooper, GS (reprint author), NIEHS, Epidemiol Branch A305, Res Triangle Pk, NC 27709 USA. OI Baird, Donna/0000-0002-5544-2653 NR 36 TC 61 Z9 62 U1 0 U2 1 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 15 PY 2001 VL 153 IS 12 BP 1159 EP 1165 DI 10.1093/aje/153.12.1159 PG 7 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 443JK UT WOS:000169337900006 PM 11415950 ER PT J AU Dunson, DB AF Dunson, DB TI Commentary: Practical advantages of Bayesian analysis of epidemiologic data SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Article DE Bayes theorem; epidemiologic methods; hierarchical Bayes; latent variable; Markov chain Monte Carlo; posterior probability; prior distribution ID LATENT VARIABLE MODELS; MEASUREMENT ERROR; BREAST-CANCER; STATISTICAL-METHODS; MEDICAL STATISTICS; P-VALUES; DISEASE; SUSCEPTIBILITY; PROBABILITIES; HYPOTHESIS AB In the past decade, there have been enormous advances in the use of Bayesian methodology for analysis of epidemiologic data, and there are now many practical advantages to the Bayesian approach. Bayesian models can easily accommodate unobserved variables such as an individual's true disease status in the presence of diagnostic error. The use of prior probability distributions represents a powerful mechanism for incorporating information from previous studies and for controlling confounding. Posterior probabilities can be used as easily interpretable alternatives to p values. Recent developments in Markov chain Monte Carlo methodology facilitate the implementation of Bayesian analyses of complex data sets containing missing observations and multidimensional outcomes. Tools are now available that allow epidemiologists to take advantage of this powerful approach to assessment of exposure-disease relations. C1 NIEHS, Biostat Branch, Res Triangle Pk, NC 27709 USA. RP Dunson, DB (reprint author), NIEHS, Biostat Branch, POB 12233,MD A3-03, Res Triangle Pk, NC 27709 USA. NR 39 TC 74 Z9 77 U1 0 U2 5 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 15 PY 2001 VL 153 IS 12 BP 1222 EP 1226 DI 10.1093/aje/153.12.1222 PG 5 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 443JK UT WOS:000169337900014 PM 11415958 ER PT J AU Daniels, JL Olshan, AF Teschke, K Hertz-Picciotto, I Savitz, DA Blatt, J AF Daniels, JL Olshan, AF Teschke, K Hertz-Picciotto, I Savitz, DA Blatt, J TI Comparison of assessment methods for pesticide exposure in a case-control interview study SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Article DE case-control studies; child; epidemiologic methods; interviews; neoplasms; neuroblastoma; occupational exposure; pesticides ID OCCUPATIONAL CASE-CONTROL; ALLOYED GOLD STANDARD; PARENTAL OCCUPATION; NEUROBLASTOMA; INFORMATION; POPULATION; CANCER; RISK AB In epidemiologic studies, much of the variation in disease risk estimates associated with occupational pesticide exposure may be due to variation in exposure classification. The authors compared five different methods of using interview information to assess occupational pesticide exposure in a US-Canada case-control study of neuroblastoma (1992-1994). For each method, exposure assignment was compared with that of a reference method, and neuroblastoma effect estimates were calculated. Compared with the reference method, which included a complete review of occupation, industry, job tasks, and exposure-specific activities, the use of occupation-industry groups alone or in combination with general job task information diluted the exposed group by including individuals who were unlikely to have been truly exposed. The effect estimates representing associations between each exposure method and neuroblastoma were different enough to influence the study's conclusions, especially when the exposure was rare (for maternal occupational pesticide exposure, the odds ratio was 0.7 using the reference exposure assessment method and 3.2 using the occupation-industry group exposure assessment method). Exposure-specific questions about work activities can help investigators distinguish truly exposed individuals from those who report exposure but are unlikely to have been exposed above background levels and from those who have not been exposed but are misclassified as exposed because of their employment in an occupation-industry group determined a priori to be exposed. C1 NIEHS, Epidemiol Branch, Res Triangle Pk, NC 27709 USA. Univ N Carolina, Sch Publ Hlth, Dept Epidemiol, Chapel Hill, NC 27514 USA. Univ British Columbia, Fac Med, Dept Hlth Care Epidemiol, Vancouver, BC V6T 1W5, Canada. Univ N Carolina, Sch Med, Div Pediat Hematol Oncol, Chapel Hill, NC 27514 USA. RP Daniels, JL (reprint author), NIEHS, Epidemiol Branch, MD A3-05,111 TW Alexander Dr, Res Triangle Pk, NC 27709 USA. FU NCI NIH HHS [CA 57004] NR 27 TC 24 Z9 26 U1 1 U2 6 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 15 PY 2001 VL 153 IS 12 BP 1227 EP 1232 DI 10.1093/aje/153.12.1227 PG 6 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 443JK UT WOS:000169337900015 PM 11415959 ER PT J AU Negishi, M Pedersen, LG Petrotchenko, E Shevtsov, S Gorokhov, A Kakuta, Y Pedersen, LC AF Negishi, M Pedersen, LG Petrotchenko, E Shevtsov, S Gorokhov, A Kakuta, Y Pedersen, LC TI Structure and function of sulfotransferases SO ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS LA English DT Review DE sulfotransferases; sulfuryl transfer; sulfation; sulfonation; steroids; glucosaminylglycans; drug metabolism ID SITE-DIRECTED MUTAGENESIS; N-DEACETYLASE/N-SULFOTRANSFERASE; HUMAN DOPAMINE SULFOTRANSFERASE; HEPARAN-SULFATE BIOSYNTHESIS; CRYSTAL-STRUCTURE; ESTROGEN SULFOTRANSFERASE; SUBSTRATE-SPECIFICITY; MUTATIONAL ANALYSIS; AMINO-ACID; SULT1A3 AB Sulfotransferases (STs) catalyze the transfer reaction of the sulfate group from the ubiquitous donor 3 ' -phosphoadenosine 5 ' -phosphosulfate (PAPS) to an acceptor group of numerous substrates, This reaction, often referred to as sulfuryl transfer, sulfation, or sulfonation, is widely observed from bacteria to humans and plays a key role in various biological processes such as cell communication, growth and development, and defense, The cytosolic STs sulfate small molecules such as steroids, bioamines, and therapeutic drugs, while the Golgi-membrane counterparts sulfate large molecules including glucosaminylglycans and proteins. We have now solved the X-ray crystal structures of four cytosolic and one membrane ST. All five STs are globular proteins composed of a single alpha/beta domain with the characteristic five-stranded beta -sheet, The beta -sheet constitutes the core of the Paps-binding and catalytic sites. Structural analysis of the PAPS-, PAP-, substrate-, and/or orthovanadate (VO43-)-bound enzymes has also revealed the common molecular mechanism of the transfer reaction catalyzed by sulfotransferses. The X-ray crystal structures have opened a new era for the study of sulfotransferases. (C) 2001 Academic Press. C1 NIEHS, Reprod & Dev Toxicol Lab, Pharmacogenet Sect, NIH, Res Triangle Pk, NC 27709 USA. NIEHS, Struct Biol Lab, NIH, Res Triangle Pk, NC 27709 USA. Univ N Carolina, Dept Chem, Chapel Hill, NC 27599 USA. Osaka Univ, Grad Sch Sci, Dept Biol, Toyonaka, Osaka 5600043, Japan. RP Negishi, M (reprint author), NIEHS, Reprod & Dev Toxicol Lab, Pharmacogenet Sect, NIH, Res Triangle Pk, NC 27709 USA. RI Pedersen, Lee/E-3405-2013 OI Pedersen, Lee/0000-0003-1262-9861 NR 45 TC 168 Z9 173 U1 1 U2 23 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 JUN 15 PY 2001 VL 390 IS 2 BP 149 EP 157 DI 10.1006/abbi.2001.2368 PG 9 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 446YY UT WOS:000169542800001 PM 11396917 ER PT J AU Chen, A Gao, ZG Barak, D Liang, BT Jacobson, KA AF Chen, A Gao, ZG Barak, D Liang, BT Jacobson, KA TI Constitutive activation of A(3) adenosine receptors by site-directed mutagenesis SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE purines; G protein-coupled receptor; phospholipase C; adenylyl cyclase; radioligand binding; nucleosides ID PROTEIN-COUPLED RECEPTORS; TERNARY COMPLEX MODEL; ALPHA(1B)-ADRENERGIC RECEPTOR; BETA(2)-ADRENERGIC RECEPTOR; PHOSPHOLIPASE-C; ANTAGONISTS; HYDROLYSIS; MUTATIONS; AGONISTS; MOTIF AB The objective of this study was to create constitutively active mutant human A, adenosine receptors (ARs) using single amino acid replacements, based on findings from other G protein-coupled receptors, A, ARs mutated in transmembrane helical domains (TMs) 1, 3, 6, and 7 were expressed in COS-7 cells and subjected to agonist radioligand binding and phospholipase C (PLC) and adenylyl cyclase (AC) assays. Three mutant receptors, A229E in TM6 and R108A and R108K in the DRY motif of TM3, were found to be constitutively active in both functional assays. The potency of the A, agonist CI-IB-MECA (1-chloro-N-6-(3-iodobenzyl)adenosine-5'-N-methyluronamide) in PLC activation was enhanced by at least an order of magnitude over wild type (EC50 951 nM) in R108A and A229E mutant receptors. CI-IB-MECA was much less potent (>10-fold) in C88F, Y109F, and Y282F and mutants or inactive following double mutation of the DRY motif. The degree of constitutive activation was more pronounced for the AC signaling pathway than for the PLC signaling pathway. The results indicated that specific locations within the TMs proximal to the cytosolic region were responsible for constraining the receptor in a G protein-uncoupled conformation. (C) 2001 Academic Press. C1 NIDDKD, Mol Recognit Sect, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. Univ Penn, Med Ctr, Dept Med, Div Cardiovasc, Philadelphia, PA 19104 USA. Univ Penn, Med Ctr, Dept Pharmacol, Philadelphia, PA 19104 USA. RP NIDDKD, Mol Recognit Sect, Bioorgan Chem Lab, NIH, Bldg 8A,Rm B1A-19, Bethesda, MD 20892 USA. EM kajacobs@helix.nih.gov RI Jacobson, Kenneth/A-1530-2009 OI Jacobson, Kenneth/0000-0001-8104-1493 FU Intramural NIH HHS [Z99 DK999999, Z01 DK031117-20]; NHLBI NIH HHS [R01 HL048225, R01-HL48225] NR 36 TC 31 Z9 31 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 0006-291X EI 1090-2104 J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD JUN 15 PY 2001 VL 284 IS 3 BP 596 EP 601 DI 10.1006/bbrc.2001.5027 PG 6 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 443WR UT WOS:000169365800009 PM 11396942 ER PT J AU Hur, GM Ryu, YS Hong, JH Bae, SH Bae, JY Paik, SG Kim, YM Seok, JH Lee, JH AF Hur, GM Ryu, YS Hong, JH Bae, SH Bae, JY Paik, SG Kim, YM Seok, JH Lee, JH TI Serum after partial hepatectomy stimulates iNOS gene transcription via downstream NF-kappa B site SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article ID NITRIC-OXIDE SYNTHASE; HEPATOCYTE GROWTH-FACTOR; LIVER-REGENERATION; INTERFERON-GAMMA; RAT; BINDING; LIPOPOLYSACCHARIDE; ENHANCEMENT; ACTIVATION; EXPRESSION AB It has been known that the expression of inducible nitric oxide synthase (iNOS) is up-regulated during hepatic regeneration. The present study characterized the molecular mechanisms involved in the transcriptional activation of iNOS gene by using the serum after partial hepatectomy (post-PH serum) in vitro. The post-PH serum rapidly induced iNOS mRNA expression, which was blocked by anti-tumor necrosis factor-alpha (TNF-alpha) antibody in BNL CL.2 cells, murine embryonic liver cell line. In addition, EMSAs using a NF-kappaB-specific oligomer showed that the up-regulated iNOS mRNA expression in cells treated with post-PH serum correlated with transient activation of NF-kappaB complex (p50/p65 heterodimer). Transient transfection of BNL CL.2 cells with iNOS promoter linked to a CAT reporter gene showed the transcriptional activation of iNOS promoter by post-PH serum. Furthermore, site-directed mutational analysis of the two MF-kappaB sites individually or in combination revealed that iNOS expression by post-PH serum is regulated by the downstream NF-kappaB site, but not by upstream NF-kappaB site. Taken together, these results suggest that the downstream NF-kappaB site acts as an essential component for the iNOS expression by post-PH serum during hepatic regeneration. (C) 2001 Academic Press. C1 Chungnam Natl Univ, Coll Med, Dept Pharmacol, Taejon 301131, South Korea. Chungnam Natl Univ, Coll Med, Dept Biol, Taejon 301131, South Korea. NINDS, Genet Pharmacol Unit, NIH, Bethesda, MD 20892 USA. RP Hur, GM (reprint author), Chungnam Natl Univ, Coll Med, Dept Pharmacol, 6 Munhwa Dong, Taejon 301131, South Korea. NR 28 TC 3 Z9 3 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD JUN 15 PY 2001 VL 284 IS 3 BP 607 EP 613 DI 10.1006/bbrc.2001.5034 PG 7 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 443WR UT WOS:000169365800011 PM 11396944 ER PT J AU Shimizu, C Fuda, H Lee, YC Strott, CA AF Shimizu, C Fuda, H Lee, YC Strott, CA TI Transcriptional regulation of human 3 '-phosphoadenosine 5 '-phosphosulfate synthase 1 SO BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS LA English DT Article DE ATP sulfurylase; APS kinase; PAPS synthase; sulfonation; sulfoconjugation ID REPRESS TRANSCRIPTION; SP-FAMILY; PROMOTER; ACTIVATION; EXPRESSION; DOMAINS; PROTEIN; GENE; RECEPTOR; COMPLEX AB Sulfonation, which is essential for normal growth, development and maintenance of the internal milieu, requires the universal sulfonate donor molecule 3'-phosphoadenosine 5'-phosphosulfate (PAPS) produced from ATP and inorganic sulfate by two bifunctional PAPS synthase isozymes. The gene for PAPS synthase 1 containing neither a TATA nor a CCAAT box was found to be under the influence off the Sp1 family of transcription factors. Multiple GC/GT boxes are present in the proximal promoter region and deletion analysis implicated their involvement in transcription, a finding supported by mutational analysis of specific GC/GT boxes. Nuclear extract of SW13 cells, which highly express PAPS synthase 1, contains proteins that bind to probes possessing specific QC/GT boxes; furthermore, the presence of Sp1, Sp2, and Sp3 proteins in nuclear extracts was confirmed by supershift analysis, Cotransfection experiments using SL2 cells yielded additional support for the involvement of Sp1 in transcriptional regulation of the PAPS synthase 1 gene; the involvement of Sp2 and/or Sp3 is presently unclear. C1 NICHD, Endocrinol & Reprod Res Branch, Sect Steroid Regulat, NIH, Bethesda, MD 20892 USA. RP Strott, CA (reprint author), NICHD, Endocrinol & Reprod Res Branch, Sect Steroid Regulat, NIH, Bldg 49,Room 6A36, Bethesda, MD 20892 USA. NR 27 TC 7 Z9 7 U1 0 U2 0 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0006-291X J9 BIOCHEM BIOPH RES CO JI Biochem. Biophys. Res. Commun. PD JUN 15 PY 2001 VL 284 IS 3 BP 763 EP 770 DI 10.1006/bbrc.2001.5032 PG 8 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 443WR UT WOS:000169365800035 PM 11396968 ER PT J AU Hyman, SE AF Hyman, SE TI Mood disorders in children and adolescents: An NIMH perspective SO BIOLOGICAL PSYCHIATRY LA English DT Editorial Material ID MANIC-DEPRESSIVE ILLNESS; HIPPOCAMPAL NEUROGENESIS; PSYCHIATRIC-DISORDERS; GENE-EXPRESSION; STRESS; BRAIN; EXPERIENCE; RECURRENT; ANXIETY; FAMILY C1 NIH, Bethesda, MD 20892 USA. RP Hyman, SE (reprint author), NIH, Bldg 10, Bethesda, MD 20892 USA. NR 37 TC 14 Z9 17 U1 2 U2 3 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0006-3223 J9 BIOL PSYCHIAT JI Biol. Psychiatry PD JUN 15 PY 2001 VL 49 IS 12 BP 962 EP 969 DI 10.1016/S0006-3223(01)01128-3 PG 8 WC Neurosciences; Psychiatry SC Neurosciences & Neurology; Psychiatry GA 447JV UT WOS:000169568900003 PM 11430838 ER PT J AU Kaufman, J Martin, A King, RA Charney, D AF Kaufman, J Martin, A King, RA Charney, D TI Are child-, adolescent-, and adult-onset depression one and the same disorder? SO BIOLOGICAL PSYCHIATRY LA English DT Review DE children; adolescents; depression; neurobiology ID DEXAMETHASONE SUPPRESSION TEST; PREPUBERTAL MAJOR DEPRESSION; GROWTH-HORMONE SECRETION; CORTICOTROPIN-RELEASING-FACTOR; THYROID STIMULATING HORMONE; CONTROLLED FAMILY HISTORY; PLACEBO-CONTROLLED TRIAL; DOUBLE-BLIND; NEUROENDOCRINE RESPONSE; CORTISOL SECRETION AB This paper reviews prior research studies examining neurobiological correlates and treatment response of depression in children, adolescents, and adults. Although theta are some similarities in research findings observed across the life cycle, both children and adolescents have been found to differ fram depressed adults on measures of basal cortisol sedation, corticotropin stimulation post-corticotropin releasing hormone (CRH) infusion, response ra several serotonergic probes immunity indices, and efficacy; of a tricyclic medications. These differences are proposed to be doe to 1) developmental factors, 2) stage of illness factors (e.g., number of episodes, total duration of illness), or 3) heterogeneity in clinical outcome (e.g., recurrent unipolar course vs. new-onset bipolar disorder). Relevant clinical and preclinical studies that provide support for these alternate explanations of the discrepant findings are reviewed, and directions for future research are discussed. To determine whether child, adolescent-, and adult-onset depression represent the same condition, it is recommended that researchers II use the same neuroimaging paradigms in child, adolescent and adult depressed cohorts; 2) carefully characterize subjects' stage of illness; and 3) conduct longitudinal clinical and repeat neurobiological assessments of patients of diffrrent ages at various stages of illness. In addition careful attention to familial subtypes (e.g., depressive spectrum disorders vs. familial pure depressive disorders) and environmental factors; (e.g., trauma history) are suggested for future investigations, (C) 2001 Society of Biological Psychiatry. C1 Yale Univ, Sch Med, Dept Psychiat, New Haven, CT 06511 USA. Yale Univ, Ctr Child Study, Dept Psychiat, New Haven, CT 06511 USA. NIMH, Bethesda, MD 20892 USA. RP Kaufman, J (reprint author), Yale Univ, Sch Med, Dept Psychiat, Univ Towers Suite 2H,100 York St, New Haven, CT 06511 USA. NR 191 TC 157 Z9 160 U1 6 U2 25 PU ELSEVIER SCIENCE INC PI NEW YORK PA 655 AVENUE OF THE AMERICAS, NEW YORK, NY 10010 USA SN 0006-3223 J9 BIOL PSYCHIAT JI Biol. Psychiatry PD JUN 15 PY 2001 VL 49 IS 12 BP 980 EP 1001 DI 10.1016/S0006-3223(01)01127-1 PG 22 WC Neurosciences; Psychiatry SC Neurosciences & Neurology; Psychiatry GA 447JV UT WOS:000169568900006 PM 11430841 ER PT J AU Chen, YD Yan, B AF Chen, YD Yan, B TI Theoretical formalism for bead movement powered by single two-headed motors in a motility assay SO BIOPHYSICAL CHEMISTRY LA English DT Article DE kinesin; dynein; free-energy transduction; microtubule; Brownian motion ID KINESIN MOLECULES; STOCHASTIC-MODEL; PROTEIN KINETICS; ATP HYDROLYSIS; FORCE; MICROTUBULES; CYCLES; LOAD AB Kinesins and dyneins are protein motors that can use the free energy of ATP hydrolysis to carry a cargo and move uni-directionally along a microtubule filament. The purpose of this paper is to derive the formalism connecting the ATP-driven translocation reactions of these motors on microtubule filaments and the movement of the bead carried by the motor in a motility assay in which the bead is clamped at an arbitrary constant force. The formalism is thus useful in elucidating the load-dependent kinetic mechanism of the free-energy transduction of the motor using the mechanical data obtained from the motility assay. The formalism is also useful in assessing the effect on the measured motility data of various physical and hydrodynamic parameters of the assay, such as the size of the bead, the viscosity of the medium, the stiffness of the elastic element connecting the motor and the bead, etc. In a previous paper [Biophys. J. 67 (2000) 313] (hereafter referred to as paper I), we have derived the formalism for the case that the motor in the assay has only one head. In this paper we extend the derivation to the case that the motor is two-headed. The formalism is derived based on a simple two-state hand-over-hand model for the movement of the motor on microtubule, but can be easily extended to more complicated kinetic models. Effects of various hydrodynamic parameters on the velocity of the bead are studied with numerical calculations of the model. The difference between the formalism presented in this paper and the widely used 'chemical' formalism, in which the movement of the kinesin and the bead is described by pure chemical reactions, is discussed. (C) 2001 Elsevier Science B.V. All rights reserved. C1 NIDDKD, Math Res Branch, NIH, Bethesda, MD 20892 USA. RP Chen, YD (reprint author), NIDDKD, Math Res Branch, NIH, Bethesda, MD 20892 USA. NR 32 TC 9 Z9 9 U1 0 U2 1 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0301-4622 J9 BIOPHYS CHEM JI Biophys. Chem. PD JUN 15 PY 2001 VL 91 IS 1 BP 79 EP 91 DI 10.1016/S0301-4622(01)00153-3 PG 13 WC Biochemistry & Molecular Biology; Biophysics; Chemistry, Physical SC Biochemistry & Molecular Biology; Biophysics; Chemistry GA 450AV UT WOS:000169720100007 PM 11403886 ER PT J AU Wang, JY Wu, JN Cherng, TL Hoffer, BJ Chen, HH Borlongan, CV Wang, Y AF Wang, JY Wu, JN Cherng, TL Hoffer, BJ Chen, HH Borlongan, CV Wang, Y TI Vitamin D-3 attenuates 6-hydroxydopamine-induced neurotoxicity in rats SO BRAIN RESEARCH LA English DT Article DE vitamin D-3; 6-hydroxydopamine; neuroprotection; dopamine; glial cell line-derived neurotrophic factor; Parkinson's disease ID MIDBRAIN DOPAMINERGIC-NEURONS; NEUROTROPHIC FACTOR PROTECTS; KAINATE-INDUCED EXCITATION; FACTOR MESSENGER-RNA; IN-VIVO; 1,25-DIHYDROXYVITAMIN D-3; GLIAL-CELLS; DIFFERENTIAL REGULATION; TYROSINE-HYDROXYLASE; NEUROBLASTOMA-CELLS AB Previous reports have demonstrated that exogeneous administration of glial cell line-derived neurotrophic factor (GDNF) reduces ventral mesencephalic (VM) dopaminergic (DA) neuron damage induced by 6-hydroxydopamine (6-OHDA) lesioning in rats. Recent studies have shown that 1,25-dihydroxyvitamin D-3 (D3) enhances endogenous GDNF expression in vitro and in vivo. The purpose of present study was to investigate if administration of D3 in vivo and in vitro would protect against 6-OHDA-induced DA neuron injury. Adult male Sprague-Dawley rats were injected daily with D3 or with saline for 8 days and then lesioned unilaterally with 6-OHDA into the medial forebrain bundle. Locomotor activity was measured using automated activity chambers. We found that unilateral 6-OHDA lesioning reduced locomotor activity in saline-pretreated animals. Pretreatment with D3 for 8 days significantly restored locomotor activity in the lesioned animals. All animals were sacrificed for neurochemical analysis 6 weeks after lesioning. We found that 6-OHDA administration significantly reduced dopamine (DA), 3,4-dihydroxy-phenylacetic acid (DOPAC) and homovanilic acid (HVA) levels in the substantia nigra (SN) on the lesioned side in the saline-treated rats. D3 pretreatment protected against 6-OHDA-mediated depletion of DA and its metabolites in SN. Using primary cultures obtained from the VM of rat embryos, we found that 6-OHDA or H2O2 alone caused significant cell death. Pretreatment with D3 (10(-10) M) protected VM neurons against 6-OHDA- or H2O2-induced cell death in vitro. Taken together, our data indicate that D3 pretreatment attenuates the hypokinesia and DA neuronal toxicity induced by 6-OHDA. Since both H2O2 and 6-OHDA may injure cells via free radical and reactive oxygen species, the neuroprotection seen here may operate via a reversal of such a toxic mechanism. Published by Elsevier Science B.V. C1 NIDA, Baltimore, MD 21224 USA. Natl Def Med Ctr, Dept Physiol, Taipei, Taiwan. Natl Def Med Ctr, Dept Pharmacol, Taipei, Taiwan. RP Hoffer, BJ (reprint author), NIDA, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. OI Borlongan, Cesar/0000-0002-2966-9782 NR 43 TC 121 Z9 128 U1 0 U2 3 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0006-8993 J9 BRAIN RES JI Brain Res. PD JUN 15 PY 2001 VL 904 IS 1 BP 67 EP 75 DI 10.1016/S0006-8993(01)02450-7 PG 9 WC Neurosciences SC Neurosciences & Neurology GA 447TQ UT WOS:000169588900007 PM 11516412 ER PT J AU Stewart, JH AF Stewart, JH TI Lung carcinoma in African Americans - A review of the current literature SO CANCER LA English DT Review ID LOS-ANGELES-COUNTY; SURGICAL ADJUVANT BREAST; CANCER RISK; GENETIC-POLYMORPHISM; CIGARETTE-SMOKING; CYP1A1 POLYMORPHISM; BOWEL PROJECT; UNITED-STATES; RACE; CAUCASIANS C1 NCI, Surg Branch, Thorac Oncol Sect, NIH, Bethesda, MD 20892 USA. RP Stewart, JH (reprint author), NCI, Surg Branch, Thorac Oncol Sect, NIH, Bldg 10,Room 2B07,10 Ctr Dr,MSC 1502, Bethesda, MD 20892 USA. FU NCI NIH HHS [K08 CA131482] NR 42 TC 29 Z9 29 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 JI Cancer PD JUN 15 PY 2001 VL 91 IS 12 BP 2476 EP 2482 DI 10.1002/1097-0142(20010615)91:12<2476::AID-CNCR1283>3.0.CO;2-Z PG 7 WC Oncology SC Oncology GA 443NX UT WOS:000169348200034 PM 11413540 ER PT J AU Luo, J Duggan, DJ Chen, YD Sauvageot, J Ewing, CM Bittner, ML Trent, JM Isaacs, WB AF Luo, J Duggan, DJ Chen, YD Sauvageot, J Ewing, CM Bittner, ML Trent, JM Isaacs, WB TI Human prostate cancer and benign prostatic hyperplasia: Molecular dissection by gene expression profiling SO CANCER RESEARCH LA English DT Article ID CDNA MICROARRAYS; DNA MICROARRAY; IDENTIFICATION AB Critical aspects of the biology and molecular basis for prostate malignancy remain poorly understood, To reveal fundamental differences between benign and malignant growth of prostate cells, we performed gene expression profiling of primary human prostate cancer and benign prostatic hyperplasia (BPH) using cDNA microarrays consisting of 6500 human genes. Frozen prostate specimens were processed to facilitate extraction of RNA from regions of tissue enriched in either benign or malignant epithelial cell growth within a given specimen. Gene expression in each of the 16 prostate cancer and nine BPH specimens was compared with a common reference to generate normalized measures for each gene across all of the samples. Using an analysis of complete pairwise comparisons of expression profiles among all of the samples, we observed clearly discernable patterns of overall gene expression that differentiated prostate cancer from BPH, Further analysis of the data identified 210 genes with statistically significant differences in expression between prostate cancer and BPH, These genes include many not recognized previously as differentially expressed in prostate cancer and BPH, including hepsin, which codes for a transmembrane serine protease, This study reveals for the first time that significant and widespread differences in gene expression patterns exist between benign and malignant growth of the prostate gland. Gene expression analysis of prostate tissues should help to disclose the molecular mechanisms underlying prostate malignant growth and identify molecular markers for diagnostic, prognostic, and therapeutic use. C1 Johns Hopkins Med Inst, James Buchanan Brady Urol Inst, Baltimore, MD 21287 USA. NHGRI, Canc Genet Branch, Bethesda, MD 20892 USA. RP Isaacs, WB (reprint author), Johns Hopkins Hosp, 115 Marburg,600 N Wolfe St, Baltimore, MD 21287 USA. FU NCI NIH HHS [CA 58236]; NIDDK NIH HHS [DK 52675] NR 22 TC 366 Z9 382 U1 1 U2 10 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 JUN 15 PY 2001 VL 61 IS 12 BP 4683 EP 4688 PG 6 WC Oncology SC Oncology GA 443ZU UT WOS:000169374100008 PM 11406537 ER PT J AU Kobayashi, H Song, YS Hoon, DSB Appella, E Celis, E AF Kobayashi, H Song, YS Hoon, DSB Appella, E Celis, E TI Tumor-reactive T helper lymphocytes recognize a promiscuous MAGE-A3 epitope presented by various major histocompatibility complex class II alleles SO CANCER RESEARCH LA English DT Article ID HUMAN GENE MAGE-3; MELANOMA PATIENTS; LIPOSOME VACCINE; CELL EPITOPES; PHASE-I; PEPTIDE; IDENTIFICATION; INDUCTION; MOLECULES; RESPONSES AB The development of effective T cell-based immunotherapy for cancer requires the identification of antigens capable of inducing both CTL, and T helper immune responses. Although CTLs will participate in the antitumor response mainly by exerting their lytic activity on the tumor cells, helper T lymphocytes wilt be critical for the induction and maintenance of the CTLs, Thus, effective subunit therapeutic vaccines should include both CTL and T helper epitopes from antigens expressed on the tumor cells, The product of the MAGE-A3 gene is an attractive candidate for tumor immunotherapy because it is expressed in the majority of melanomas and in a great proportion of other solid tumors. Although numerous CTL epitopes for the MAGE-A3 antigen have been reported, only a few have been described for helper T cells. Here we show that a synthetic peptide derived from the MAGE-A3 sequence (MAGE-A3(146-160)) was effective in inducing in vitro T helper responses in the context of HLA-DR1 and HLA-DR7 alleles, Most significantly, the peptide-reactive helper T lymphocytes were capable of recognizing various forms of MAGE-A3 antigen (tumor cell lysates, dead/apoptotic tumor cells, or recombinant MAGE-A3 protein), indicating that the T-cell epitope represented by peptide MAGE-A3(146-160) is naturally processed by antigen-presenting cells. These studies are relevant for the design of multi-epitope vaccines for treating MAGE-A3-expressing tumors through the simultaneous stimulation of CTL and T helper lymphocytes. C1 Mayo Clin & Mayo Fdn, Dept Immunol, Mayo Grad Sch, Rochester, MN 55905 USA. Mayo Clin & Mayo Fdn, Mayo Canc Ctr, Rochester, MN 55905 USA. NCI, NIH, Bethesda, MD 20892 USA. John Wayne Canc Inst, Santa Monica, CA 90404 USA. RP Celis, E (reprint author), Mayo Clin & Mayo Fdn, Dept Immunol, Mayo Grad Sch, GU421A, Rochester, MN 55905 USA. OI Hoon, Dave/0000-0003-1915-3683 FU NCI NIH HHS [CA80782, CA12582, CA82677]; NCRR NIH HHS [RR-00585] NR 29 TC 61 Z9 62 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 JUN 15 PY 2001 VL 61 IS 12 BP 4773 EP 4778 PG 6 WC Oncology SC Oncology GA 443ZU UT WOS:000169374100022 PM 11406551 ER PT J AU Dong, G Loukinova, E Chen, Z Gangi, L Chanturita, TI Liu, ET Van Waes, C AF Dong, G Loukinova, E Chen, Z Gangi, L Chanturita, TI Liu, ET Van Waes, C TI Molecular profiling of transformed and metastatic murine squamous carcinoma cells by differential display and cDNA microarray reveals altered expression of multiple genes related to growth, apoptosis, angiogenesis, and the NF-kappa B signal pathway SO CANCER RESEARCH LA English DT Article ID PROINFLAMMATORY CYTOKINE EXPRESSION; ANCHORAGE-INDEPENDENT GROWTH; COLONY-STIMULATING FACTOR; NUCLEAR-FACTOR; HUMAN HEAD; IN-VIVO; TUMOR-GROWTH; NECK-CANCER; CONSTITUTIVE ACTIVATION; TRANSCRIPTION FACTORS AB To identify changes in gene expression with transformation and metastasis, we investigated differential gene expression in a squamous carcinoma model established in syngeneic mice, We used mRNA differential display (DD) to detect global differences and cDNA arrays enriched for cancer-associated genes using mRNA from primary keratinocytes, transformed Pam 212 squamous carcinoma cells, and metastases of Pam 212, After DD, 72 candidate cDNAs expressed primarily in transformed and metastatic cells were selected and cloned, Fifty-seven were detected, and 32 were confirmed to be differentially expressed by Northern blot analysis. mRNA expression profiles were also generated using a mouse cDNA array composed of 4000 elements representing known genes and expressed sequence tags plus the 57 DD candidate cDNAs detected by Northern analysis to facilitate data validation. cDNA array detected 76.9% of the differentially expressed mRNAs selected from DD and confirmed by Northern blot, whereas low-abundance mRNAs did not reach the threshold for detection by the lower-sensitivity array method. Clustering analysis of Do and array results from transformed and metastatic cells identified genes that exhibited decreased or increased expression with transformation and metastasis, Alterations in the expression of several genes detected during tumor progression were consistent with their functional activities involving growth (p21, p27, and cyclin DI), resistance and apoptosis (glutathione-S-transferase, cIAP-1, PEA-15, and Fas ligand), inflammation and angiogenesis [chemokine growth-regulated oncogene 1 (also called KC)], and signal transduction (c-Met, yes-associated protein, and syk). Strikingly, 10 of 22 genes in the cluster expressed in metastases have been associated with activation of the nuclear factor (NF)-kappaB signal pathway. The NF-kappaB-inducible cytokine Gro-1 was recently shown to promote tumor growth, metastasis, and angiogenesis of squamous cell carcinomas in vivo (Loukinova et at, Oncogene, 19: 3477-3486, 2000), The results demonstrate that early response genes related to NF-kappaB contribute to metastatic tumor progression. Comparison of cell lines and tumor tissue revealed a concordance of similar to 50% by array, and 70% for Northern-confirmed, metastasis-related genes. Functional genomic approaches comparing expression among cell lines and tumor tissue may promote a better understanding of the genes expressed by malignant and host cells during tumor progression and metastasis. C1 Natl Inst Deafness & Commun Disorders, Head & Neck Surg Branch, NIH, Bethesda, MD 20892 USA. NCI, Div Clin Sci, Ctr Adv Technol, Gaithersburg, MD 20887 USA. NCI, Microarray Facil, Ctr Adv Technol, Gaithersburg, MD 20887 USA. RP Van Waes, C (reprint author), Natl Inst Deafness & Commun Disorders, Head & Neck Surg Branch, NIH, Bldg 10,Room 5D55, Bethesda, MD 20892 USA. RI Liu, Edison/C-4141-2008 FU NIDCD NIH HHS [DC-00016] NR 87 TC 110 Z9 116 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 JUN 15 PY 2001 VL 61 IS 12 BP 4797 EP 4808 PG 12 WC Oncology SC Oncology GA 443ZU UT WOS:000169374100026 PM 11406555 ER PT J AU Poulaki, V Mitsiades, N Romero, ME Tsokos, M AF Poulaki, V Mitsiades, N Romero, ME Tsokos, M TI Fas-mediated apoptosis in neuroblastoma requires mitochondrial activation and is inhibited by FLICE inhibitor protein and bcl-2 SO CANCER RESEARCH LA English DT Article ID DRUG-INDUCED APOPTOSIS; NON-HODGKINS-LYMPHOMA; HUMAN NEURO-BLASTOMA; INDUCED CELL-DEATH; CYTOCHROME-C; SCID MICE; SIGNALING COMPLEX; FAMILY PROTEINS; TUMOR-CELLS; MYC GENE AB Fas-mediated apoptosis proceeds though mitochondria-dependent or -independent pathways and is deficient in drug-resistant cells. Neuroblastoma, a common pediatric malignancy, often develops drug-resistance and has a silenced caspase 8 (FLICE) gene, which has been associated with Fas- and drug-resistance. We report that besides caspase 8, which was absent in approximately one-third of 26 neuroblastoma eases in this study, other proteins such as bcl-2 and FLICE-inhibitory protein (FLIP), are equally important in conferring Fas-resistance to neuroblastoma cells, Both bcl-2 and FLIP were frequently expressed in neuroblastoma tissues. Our in vitro studies showed that FLIP was recruited to the death-inducing signaling complex and interfered with the recruitment of caspase 8 in neuroblastoma cells. bcl-2 inhibited the activation of the mitochondria; but it also lowered the free cytoplasmic levels of caspase 8 by binding and sequestering it, thus acting through a novel antiapoptotic mechanism upstream of the mitochondria. In vitro down-regulation of bcl-2 with antisense oligonucleotides allowed the release of cytochrome e from mitochondria and the activation of caspases 8 and 3 upon Fas activation as well as sensitized neuroblastoma cells to Fas-mediated apoptosis, Downregulation of FLIP had only a modest apoptotic effect because of the coexistent mitochondrial block. However, combined treatment with bcl-2 and FLIP antisense oligonucleotides had a statistically significant synergistic effect reversing Fas-resistance in neuroblastoma cells in vitro. These data indicate that Fas-mediated apoptosis in neuroblastoma cells is mitochondria-dependent and inhibited both at the mitochondrial level and at the level of caspase 8 activation. Thus, gene-targeting therapies fur bcl-2 and FLIP may reverse Fas-resistance and prove useful in the treatment of drug-resistant neuroblastomas. C1 NIH, Pathol Lab, NCI, Bethesda, MD 20892 USA. RP Tsokos, M (reprint author), NIH, Pathol Lab, NCI, Bldg 10,Room 2A-10, Bethesda, MD 20892 USA. NR 71 TC 70 Z9 71 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 JUN 15 PY 2001 VL 61 IS 12 BP 4864 EP 4872 PG 9 WC Oncology SC Oncology GA 443ZU UT WOS:000169374100035 PM 11406564 ER PT J AU Cuello, M Ettenberg, SA Clark, AS Keane, MM Posner, RH Nau, MM Dennis, PA Lipkowitz, S AF Cuello, M Ettenberg, SA Clark, AS Keane, MM Posner, RH Nau, MM Dennis, PA Lipkowitz, S TI Down-regulation of the erbB-2 receptor by trastuzumab (herceptin) enhances tumor necrosis factor-related apoptosis-inducing ligand-mediated apoptosis in breast and ovarian cancer cell lines that overexpress erbB-2 SO CANCER RESEARCH LA English DT Article ID TRAIL-INDUCED APOPTOSIS; NF-KAPPA-B; MAMMARY EPITHELIAL-CELLS; MONOCLONAL-ANTIBODY; DEATH RECEPTORS; CARCINOMA-CELLS; TRANSCRIPTION FACTORS; DECOY RECEPTORS; KINASE-B; IN-VIVO AB We investigated whether combined treatment with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and trastuzumab could enhance the specific killing of cells that overexpress the erbB-2 receptor. The combination resulted in an enhancement of TRAIL-mediated apoptosis in all cell lines overexpressing erbB-2 receptor compared with either reagent alone. In contrast, there was no effect in cell lines with low levels of the erb-B2 receptor. Trastuzumab treatment resulted in downregulation of the erbB-2 receptor in all erbs-2-overexpressing cell lines. Similar enhancement of TRAIL toxicity was observed when the erbB-2 receptor was down-regulated using antisense oligodeoxynucleotides. Down-regulation of the erbB-2 receptor protein by trastuzumab or antisense oligodeoxynucleotides decreased Akt kinase activation but not mitogen-activated protein kinase activation. Down-regulation of Akt kinase activity by a phosphatidylinositol 3 ' -kinase inhibitor (LY294002) also resulted in enhancement of TRAIL-mediated apoptosis, Expression of a constitutively active form of Akt kinase in an erbB-2-overexpressing cell line completely abrogated the increase in TRAIL-mediated apoptosis by trastuzumab and significantly reduced the biological effect of either reagent alone. Therefore, down-regulation of the erbB-2 receptor by trastuzumab enhances TRAIL-mediated apoptosis by inhibiting Akt kinase activity. These data suggest that the combination of trastuzumab and TRAIL may allow enhanced therapeutic efficacy and specificity in the treatment of erbB-2-overexpressing tumors. C1 NCI, Dept Genet, Med Branch, Natl Naval Med Ctr, Bethesda, MD 20889 USA. NCI, Dev Therapeut Dept, Med Branch, Natl Naval Med Ctr, Bethesda, MD 20889 USA. Uniformed Serv Univ Hlth Sci, Cell & Mol Biol Program, Bethesda, MD 20889 USA. Pontificia Univ Catolica Chile, Fac Med, Dept Obstet & Gynecol, Santiago, Chile. RP Lipkowitz, S (reprint author), NCI, Dept Genet, Med Branch, Natl Naval Med Ctr, Bldg 8,Room 5101, Bethesda, MD 20889 USA. NR 68 TC 237 Z9 242 U1 0 U2 11 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 JUN 15 PY 2001 VL 61 IS 12 BP 4892 EP 4900 PG 9 WC Oncology SC Oncology GA 443ZU UT WOS:000169374100039 PM 11406568 ER PT J AU Sereti, I Lane, HC AF Sereti, I Lane, HC TI Immunopathogenesis of human immunodeficiency virus: Implications for immune-based therapies SO CLINICAL INFECTIOUS DISEASES LA English DT Review ID ACTIVE ANTIRETROVIRAL THERAPY; COLONY-STIMULATING FACTOR; PLACEBO-CONTROLLED TRIAL; CYTOTOXIC T-LYMPHOCYTES; HIV-INFECTED PATIENTS; RANDOMIZED CONTROLLED TRIAL; CD4 CELL COUNTS; RECOMBINANT HUMAN INTERLEUKIN-12; SUSTAINED VIRAL SUPPRESSION; PHASE-I TRIAL AB Human immunodeficiency virus (HIV) infection leads to a state of CD4 lymphopenia and generalized immune activation with subsequent development of opportunistic infections and neoplasms. The use of highly active antiretroviral treatment has dramatically improved the clinical outcome for HIV-infected patients, but the associated cost and toxicity and the eventual development of drug resistance have underscored the need for additional therapeutic strategies. Immune-based therapies, such as treatment with cytokines or immunosuppressants, adoptive immunotherapy, and therapeutic immunizations, are being intensely investigated as potential supplements to antiretroviral therapy. Although much data have been generated as a result of these efforts, to date there has been little evidence of the clinical efficacy of these strategies. Randomized clinical studies remain critical in evaluating the clinical significance and the role of immune-based therapies in the therapeutic armamentarium against HIV. C1 NIAID, Cellular & Mol Retrovirol Sect, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. RP Lane, HC (reprint author), NIAID, Clin & Mol Retrovirol Sect, Immunoregulat Lab, NIH, Bldg 10,Rm 11S-231,10 Ctr Dr,MSC 1876, Bethesda, MD 20892 USA. NR 158 TC 25 Z9 26 U1 0 U2 0 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 1058-4838 J9 CLIN INFECT DIS JI Clin. Infect. Dis. PD JUN 15 PY 2001 VL 32 IS 12 BP 1738 EP 1755 DI 10.1086/320758 PG 18 WC Immunology; Infectious Diseases; Microbiology SC Immunology; Infectious Diseases; Microbiology GA 436WT UT WOS:000168959400017 PM 11360217 ER PT J AU Chaconas, G Stewart, PE Tilly, K Bono, JL Rosa, P AF Chaconas, G Stewart, PE Tilly, K Bono, JL Rosa, P TI Telomere resolution in the Lyme disease spirochete SO EMBO JOURNAL LA English DT Article DE Borrelia; DNA replication; hairpin DNA; linear DNA; telomere resolution ID BACTERIUM BORRELIA-BURGDORFERI; LINEAR PLASMID TELOMERES; COVALENTLY CLOSED ENDS; RESOLVING ENZYMES; CIRCULAR PLASMIDS; HAIRPIN FORMATION; DNA; RECOMBINATION; REPLICATION; SEQUENCE AB The genus Borrelia includes the causative agents of Lyme disease and relapsing fever. An unusual feature of these bacteria is a genome that includes linear DNA molecules with covalently closed hairpin ends referred to as telomeres, We have investigated the mechanism by which the hairpin telomeres are processed during replication. A synthetic 140 bp sequence having the predicted structure of a replicated telomere was shown to function as a viable substrate for telomere resolution in vivo, and was sufficient to convert a circular replicon to a linear form. Our results suggest that the final step in the replication of linear Borrelia replicons is a site-specific DNA breakage and reunion event to regenerate covalently closed hairpin ends. The telomere substrate described here will be valuable both for in vivo manipulation of linear DNA in Borrelia and for in vitro studies to identify and characterize the telomere resolvase. C1 Univ Western Ontario, Dept Biochem, London, ON N6A 5C1, Canada. NIAID, Lab Human Bacterial Pathogenesis, Rocky Mt Labs, NIH, Hamilton, MT 59840 USA. RP Chaconas, G (reprint author), Univ Western Ontario, Dept Biochem, London, ON N6A 5C1, Canada. NR 40 TC 64 Z9 65 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 JUN 15 PY 2001 VL 20 IS 12 BP 3229 EP 3237 DI 10.1093/emboj/20.12.3229 PG 9 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 445GP UT WOS:000169450000025 PM 11406599 ER PT J AU Caffrey, JJ Darden, T Wenk, MR Shears, SB AF Caffrey, JJ Darden, T Wenk, MR Shears, SB TI Expanding coincident signaling by PTEN through its inositol 1,3,4,5,6-pentakisphosphate 3-phosphatase activity SO FEBS LETTERS LA English DT Article DE PTEN; phosphatidylinositol 3,3,5-trisphosphate; inositol 1,3,4,5,6-pentakisphosphate; 3-phosphatase; inositol phosphate ID PHOSPHATASE-ACTIVITY; TUMOR-SUPPRESSOR; CELLS; 1,3,4,6-TETRAKISPHOSPHATE; KINASE AB PTEN, a tumor suppressor among the most commonly mutated proteins in human cancer, is recognized to be both a protein phosphatase and a phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P-3) 3-phosphatase. Previous work [Maehama and Dixon, J. Biol, Chem. 273 (1998) 13375-13378] has led to a consensus that inositol phosphates are not physiologically relevant substrates for PTEN. In contrast, ne demonstrate that PTEN is an active inositol 1,3,4,5,6-penta-kisphosphate (Ins(1,3,4,5,6)P-5) 3-phosphatase when expressed and purified from bacteria or HEK cells, Kinetic data indicate Ins(1,3,3,5,6)P-5 (K-m = 7.1 muM) and PtdIns(3,4,5)P-3 (K-m = 26 muM) compete for PTEN in vivo. Transient transfection of HEK cells with PTEN decreased Ins(1,3,4,5,6)Ps levels. We discuss the physiological significance of these studies in relation to recent work showing that dephosphorylation of Ins(1,3,4,5,6)P-5 to inositol 1,4,5,6-tetrakisphosphate is a cell signaling event. (C) 2001 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved. C1 NIEHS, Lab Signal Transduct, Res Triangle Pk, NC 27709 USA. NIEHS, Struct Biol Lab, Res Triangle Pk, NC 27709 USA. Yale Univ, Sch Med, Dept Cell Biol, New Haven, CT 06510 USA. RP Shears, SB (reprint author), NIEHS, Lab Signal Transduct, POB 12233, Res Triangle Pk, NC 27709 USA. RI Wenk, Markus/D-1441-2014 NR 26 TC 26 Z9 27 U1 1 U2 2 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0014-5793 J9 FEBS LETT JI FEBS Lett. PD JUN 15 PY 2001 VL 499 IS 1-2 BP 6 EP 10 DI 10.1016/S0014-5793(01)02500-5 PG 5 WC Biochemistry & Molecular Biology; Biophysics; Cell Biology SC Biochemistry & Molecular Biology; Biophysics; Cell Biology GA 447BR UT WOS:000169549100002 PM 11418101 ER PT J AU Le Voyer, T Rouse, J Lu, ZC Lifsted, T Williams, M Hunter, KW AF Le Voyer, T Rouse, J Lu, ZC Lifsted, T Williams, M Hunter, KW TI Three loci modify growth of a transgene-induced mammary tumor: Suppression of proliferation associated with decreased microvessel density SO GENOMICS LA English DT Article ID BREAST-CANCER; MITOTIC INDEXES; MOUSE; MICE; ANGIOGENESIS; PROGRESSION; CARCINOMA; HETEROZYGOSITY; INHIBITOR; IDENTIFICATION AB In earlier studies it was observed that the genetic background significantly affected the phenotype of a transgene-induced mammary tumor. Tumors arising in an (I/LnJ x PyMT) F-1 hybrid background appeared earlier than in the FVB/N-TgN(MMTV-PyVT)(634Mul) parent, but accumulated less tumor mass, indicating a net decrease in tumor growth. Quantitative genetic mapping in a backcross identified three loci that were associated with the decreased proliferative capacity of the I/LnJ F-1 tumors. Molecular analysis of the tumors suggests that these loci may act by restricting the tumor's ability to recruit microvessels. The three loci, designated Mmtg1-3, are unlinked to the angiogenic genes Pgf2 FLt1, Flk4, Flk1, Vegf; and Vegfc, as well as the precursors of the endogenous antiangiogenic molecules angiostatin and endostatin. The Mmtg loci may therefore provide novel targets for antiangiogenic therapeutic strategies. C1 Fox Chase Canc Ctr, Philadelphia, PA 19111 USA. NCI, DCEG, Lab Populat Genet, NIH, Bethesda, MD 20892 USA. RP Hunter, KW (reprint author), 41-D702,41 Ctr Dr, Bethesda, MD 20892 USA. FU NCI NIH HHS [CA06927] NR 38 TC 25 Z9 25 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 JUN 15 PY 2001 VL 74 IS 3 BP 253 EP 261 DI 10.1006/geno.2001.6562 PG 9 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA 449EJ UT WOS:000169671100001 PM 11414753 ER PT J AU Sidjanin, DJ Parker-Wilson, DM Neuhauser-Klaus, A Pretsch, W Favor, J Deen, PMT Ohtaka-Maruyama, C Lu, Y Bragin, A Skach, WR Chepelinsky, AB Grimes, PA Stambolian, DE AF Sidjanin, DJ Parker-Wilson, DM Neuhauser-Klaus, A Pretsch, W Favor, J Deen, PMT Ohtaka-Maruyama, C Lu, Y Bragin, A Skach, WR Chepelinsky, AB Grimes, PA Stambolian, DE TI A 76-bp deletion in the Mip gene causes autosomal dominant cataract in Hfi mice SO GENOMICS LA English DT Article ID ZONULAR PULVERULENT CATARACT; CONGENITAL CATARACT; CRYSTALLIN GENE; MISSENSE MUTATION; LENS; WATER; PROTEIN; CONSTRUCTION; AQUAPORIN; FAMILIES AB Hfi is a dominant cataract mutation where heterozygotes show hydropic lens fibers and homozygotes show total lens opacity. The Hfi locus was mapped to the distal part of mouse chromosome 10 close to the major intrinsic protein (Mip), which is expressed only in cell membranes of lens fibers. Molecular analysis of Mip revealed a 76-bp deletion that resulted in exon 2 skipping in Mip mRNA, In Hfi/Hfi this deletion resulted in a complete absence of the wildtype Mip. In contrast, Hfi/+ animals had the same amount of wildtype Mip as +/+. Results from pulse-chase expression studies excluded hetero-oligomerization of wildtype and mutant Mip as a possible mechanism for cataract formation in the Hfi/+. We propose that the cataract phenotype in the Hfi heterozygote mutant is due to a detrimental gain of function by the mutant Mip resulting in either cytotoxicity or disruption in processing of other proteins important for the lens. Cataract formation in the Hfi/Hfi mouse is probably a combined result of both the complete loss of wildtype Mip and a gain of function of the mutant Mip. (C) 2001 Academic Press. C1 Univ Penn, Dept Ophthalmol, Philadelphia, PA 19104 USA. Univ Penn, Dept Mol & Cellular Engn, Philadelphia, PA 19104 USA. NEI, NIH, Bethesda, MD 20892 USA. GSF, Natl Res Ctr Environm & Hlth, Inst Mammalian Genet, Neuherberg, Germany. Univ Nijmegen, Dept Cell Physiol, Nijmegen, Netherlands. RP Stambolian, DE (reprint author), Univ Penn, Dept Ophthalmol, Philadelphia, PA 19104 USA. RI Deen, P.M.T./H-8023-2014 OI Deen, P.M.T./0000-0002-7868-4655 FU NEI NIH HHS [5T32EY07131, EY10321]; NIGMS NIH HHS [GM53457] NR 36 TC 29 Z9 30 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 JUN 15 PY 2001 VL 74 IS 3 BP 313 EP 319 DI 10.1006/geno.2001.6509 PG 7 WC Biotechnology & Applied Microbiology; Genetics & Heredity SC Biotechnology & Applied Microbiology; Genetics & Heredity GA 449EJ UT WOS:000169671100007 PM 11414759 ER PT J AU Rozemuller, H Chowdhury, PS Pastan, I Kreitman, RJ AF Rozemuller, H Chowdhury, PS Pastan, I Kreitman, RJ TI Isolation of new anti-CD30 scFvs from DNA-immunized mice by phage display and biologic activity of recombinant immunotoxins produced by fusion with truncated pseudomonas exotoxin SO INTERNATIONAL JOURNAL OF CANCER LA English DT Article DE Hodgkin's disease; monoclonal antibody; recombinant immunotoxin; anti-tumor activity; phage panning ID SINGLE-CHAIN IMMUNOTOXIN; B-CELL LYMPHOMA; ANTITUMOR-ACTIVITY; IN-VITRO; COMPLETE REGRESSION; CYTOTOXIC ACTIVITY; HUMAN CARCINOMA; HODGKINS-DISEASE; DISULFIDE BONDS; MALIGNANT-CELLS AB To target CD30 on Hodgkin's disease and anaplastic large-cell lymphoma, anti-CD30 single-chain antibodies were obtained by DNA immunization of mice with the complete human CD30 cDNA, Spleens were isolated from mice with high anti-CD30 titer, and the RNA was used for the production of an scFv-displaying phage library, Specific phages were enriched by 3 rounds of panning on soluble CD30 or CD30(+) K562 cells. Recombinant immunotoxins (rITs) were made from 3 ELISA-positive scFv phages by fusion to a 38 kDa truncated mutant of Pseudomonas exotoxin (PE38) with or without a KDEL mutant sequence at the t terminus. In vitro cytotoxicity of purified anti-CD30 rITs was measured on CD30-transfected A431 cells. IC50 values ranged from 3 to 7 ng/ml (50-110 pM) for PE38 rITs and 0.1 ng/ml (2 pM) for the PE38-KDEL IT on A431-CD30 cells, The parental A431 cells were resistant, indicating that the cytotoxicity was specific and CD30-mediated, rITs were tested for anti-tumor activity in a nude mouse model. A431-CD30 cells were injected s.c. on day 0; then, mice bearing measurable tumors were treated beginning on day 4 with 3 alternate daily doses i.v. Anti-tumor activity was dose-dependent and not found when irrelevant: ITs were administered or when CD30(-) tumors were treated, Our data show that DNA immunization and antibody phage display may be useful in producing new rITs against hematologic malignancies. Published 2001 Wiley-Liss. Inc.(dagger) C1 NCI, Mol Biol Lab, NIH, Bethesda, MD 20892 USA. RP Kreitman, RJ (reprint author), NCI, Mol Biol Lab, NIH, 37-4B27,37 Convent Dr MSC 4255, Bethesda, MD 20892 USA. NR 42 TC 17 Z9 17 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 JUN 15 PY 2001 VL 92 IS 6 BP 861 EP 870 DI 10.1002/ijc.1266 PG 10 WC Oncology SC Oncology GA 434UJ UT WOS:000168834000014 PM 11351308 ER PT J AU Masumi, A Ozato, K AF Masumi, A Ozato, K TI Coactivator p300 acetylates the interferon regulatory factor-2 in U937 cells following phorbol ester treatment SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HISTONE ACETYLATION; DNA-BINDING; GENE-TRANSCRIPTION; ONCOPROTEIN E1A; FACTOR-II; PCAF; PHOSPHORYLATION; ACTIVATION; CBP; ACETYLTRANSFERASE AB Interferon regulatory factor-2 (IRF-2) is at transcription factor of the IRF family that represses interferon-mediated gene expression. In the present study, we show that human monocytic U937 cells express truncated forms of IRF-2 containing the DNA binding do main but lacking much of the C-terminal regulatory domain. U937 cells are shown to respond to phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA) to induce expression of histone acetylases p300 and p300/CBP-associated factor (PCAF), In addition, TPA treatment led to the appearance of full-length IRF-2, along with a reduction of the truncated protein. interestingly, full-length IRF-2 in TPA-treated U937 cells occurred as a complex with p300 as well as PCAF and was itself acetylated. Consistent with these results, recombinant IRF-2 was acetylated by p300 and to a lesser degree by PCAF in vitro. Another IRF member, IRF-1, an activator of interferon-mediated transcription, was also acetylated in vitro by these acetylases, Finally, we demonstrate that the addition of IRF-2 but not IRF-1. inhibits core histone acetylation by p300 in vitro, The addition of IRF-2 also inhibited acetylation of nucleosomal histones in TPA-treated U937 cells. Acetylated IRF-2 may affect local chromatin structure in vivo by inhibiting core histone acetylation and may serve as a mechanism by which IRF-2 negatively regulates interferon-inducible transcription. C1 Natl Inst Infect Dis, Dept Safety Res Biol, Tokyo 2080011, Japan. NICHD, Lab Mol Growth Regulat, NIH, Bethesda, MD 20892 USA. RP Masumi, A (reprint author), Natl Inst Infect Dis, Dept Safety Res Biol, 4-7-1 Gakuen,Musashimurayama Shi, Tokyo 2080011, Japan. NR 51 TC 41 Z9 42 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 15 PY 2001 VL 276 IS 24 BP 20973 EP 20980 DI 10.1074/jbc.M101707200 PG 8 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 442RC UT WOS:000169297900025 PM 11304541 ER PT J AU Sauna, ZE Smith, MM Muller, M Ambudkar, SV AF Sauna, ZE Smith, MM Muller, M Ambudkar, SV TI Functionally similar vanadate-induced 8-azidoadenosine 5 '-[alpha-P-32]diphosphate-trapped transition state intermediates of human P-glycoprotein are generated in the absence and presence of ATP hydrolysis SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID NUCLEOTIDE-BINDING DOMAINS; REPAIR PROTEIN MUTS; HAMSTER OVARY CELLS; MULTIDRUG TRANSPORTER; CRYSTAL-STRUCTURE; CATALYTIC SITES; RESISTANCE PUMP; ABC TRANSPORTER; WALKER-A; SUBUNIT AB P-glycoprotein (Pgp) is an ATP-dependent drug efflux pump whose overexpression confers multidrug resistance to cancer cells. Pgp exhibits a robust drug substrate-stimulable ATPase activity, and vanadate (Vi) blocks this activity effectively by trapping Pgp nucleotide in a noncovalent stable transition state conformation. In this study we compare Vi-induced [alpha-P-32]8-azido-ADP trapping into Pgp in the presence of [alpha-P-32]8-azido-ATP (with ATP hydrolysis) or [alpha-P-32]8-azido-ADP (without ATP hydrolysis), Vi mimics Pi to trap the nucleotide tenaciously in the Pgp . [alpha-P-32]8-azido-ADP.Vi conformation in either condition. Thus, by using [alpha-P-32]8-azido-ADP we show that the Vi-induced transition state of Pgp can be generated even in the absence of ATP hydrolysis. Furthermore, half-maximal trapping of nucleotide into Pgp in the]presence of Vi occurs at similar concentrations of [(alpha-P-32]8-azido-ATP or [alpha-P-32]8-azido-ADP. The trapped [(alpha-P-32]8-azido-ADP is almost equally distributed between the N- and the C-terminal ATP sites of Pgp in both conditions. Additionally, point mutations in the Walker B domain of either the N- (D555N) or C (D1200N)-terminal ATP sites that arrest ATP hydrolysis and Vi-induced trapping also show abrogation of [alpha-P-32]8-azido-ADP trapping into Pgp in the absence of hydrolysis, These data suggest that both ATP sites are dependent on each other for function and that each site exhibits similar affinity for 8-azido-ATP (ATP) or 8-azido-ADP (ADP), Similarly, Pgp in the transition state conformation generated with either ADP or ATP exhibits drastically reduced affinity for the binding of analogues of drug substrate ([I-125]iodoarylazidoprazosin) as well as nucleotide (2'(3')-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate). Analyses of Arrhenius plots show that trapping of Pgp with [alpha-P-32]8-azido-ADP tin the, absence of hydrolysis) displays an similar to2.5-fold higher energy of activation (152 kJ/mol) compared with that observed when the transition state intermediate is generated through hydrolysis of [alpha-P-32]8-azido-ATP (62 kJ/mol), In aggregate, these results demonstrate that the Pgp [alpha-P-32]8-azido-ADP (or ADP) Vi transition state complexes generated either in the absence of or accompanying [alpha-P-32]8-azido-ATP hydrolysis are functionally indistinguishable. C1 NCI, Cell Biol Lab, Ctr Canc Res, NIH, Bethesda, MD 20892 USA. RP Ambudkar, SV (reprint author), NCI, Cell Biol Lab, Ctr Canc Res, NIH, Bldg 37,Rm 1B-22,37 convent Dr, Bethesda, MD 20892 USA. RI Ambudkar, Suresh/B-5964-2008 NR 44 TC 34 Z9 34 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 JUN 15 PY 2001 VL 276 IS 24 BP 21199 EP 21208 DI 10.1074/jbc.M100886200 PG 10 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 442RC UT WOS:000169297900055 PM 11287418 ER PT J AU Skeel, A Leonard, EJ AF Skeel, A Leonard, EJ TI alpha(1)-antichymotrypsin is the human plasma inhibitor of macrophage ectoenzymes that cleave pro-macrophage stimulating protein SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID AMINO-ACID-SEQUENCE; HUMAN CATHEPSIN-G; TYROSINE KINASE; PROTEOLYTIC CLEAVAGE; GENE-PRODUCT; RECEPTOR; ACTIVATION; EXPRESSION; ALPHA-1-ANTICHYMOTRYPSIN; IDENTIFICATION AB Macrophage stimulating protein (MSP) is secreted as 78-kDa single chain pro-MSP, which is converted to biologically active, disulfide-linked alpha beta chain MSP by cleavage at Arg(483)-Val(484) Murine resident peritoneal macrophages have two cell surface proteolytic activities that cleave pro-MSP. One is a pro-MSP convertase, which cleaves pro-MSP to active MSP; the other degrades pro-MSP, The degrading protease is inhibited by soybean trypsin inhibitor or by low concentrations of blood plasma, which allows the convertase to cleave pro-MSP to MSP. Using pro-MSP cleavage as the assay, we purified the inhibitor from human plasma. The bulk of the plasma protein was removed by salting out and by isoelectric precipitation of albumin. Highly purified inhibitor was then obtained in three steps: dye-ligand binding and elution, ion exchange chromatography, and high performance liquid chromatography gel filtration. After SDS-polyacrylamide gel electrophoresis and transfer to a polyvinylidene membrane, N-terminal sequencing of the product identified it as alpha (1)-antichymotrypsin, The mean concentration of alpha (1)-antichymotrypsin in human plasma is 7 muM. At this concentration, alpha (1)-antichymotrypsin inhibits both macrophage enzymes. A concentration of 0.4 muM, which is in the expected concentration range in extracellular fluid, preferentially inhibits the degrading enzyme, which allows for cleavage to active MSP by the pro-MSP convertase. C1 NCI, Immunopathol Sect, Immunobiol Lab, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RP Leonard, EJ (reprint author), NCI, Immunopathol Sect, Immunobiol Lab, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. EM leonarde@mail.ncifcrf.gov NR 30 TC 9 Z9 11 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 15 PY 2001 VL 276 IS 24 BP 21932 EP 21937 DI 10.1074/jbc.M100652200 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 442RC UT WOS:000169297900147 PM 11274154 ER PT J AU Shackelford, RE Innes, CL Sieber, SO Heinloth, AN Leadon, SA Paules, RS AF Shackelford, RE Innes, CL Sieber, SO Heinloth, AN Leadon, SA Paules, RS TI The Ataxia telangiectasia gene product is required for oxidative stress-induced G(1) and G(2) checkpoint function in human fibroblasts SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID DNA-DAMAGE; XERODERMA-PIGMENTOSUM; IONIZING-RADIATION; HYDROGEN-PEROXIDE; SKIN FIBROBLASTS; CELL; ATM; P53; REPAIR; CANCER AB Ataxia telangiectasia (AT) is an autosomal recessive disorder characterized by neuronal degeneration accompanied by ataxia, telangiectasias, acute cancer predisposition, and sensitivity to ionizing radiation (IR). Cells from individuals with AT show unusual sensitivity to IR, severely attenuated cell cycle checkpoint functions, and poor p53 induction in response to IR compared with normal human fibroblasts (NHFs), The gene mutated in AT (ATM) has been cloned, and its product, pATM, has IR-inducible kinase activity. The AT phenotype has been suggested to be a consequence, at least in part, of an inability to respond appropriately to oxidative damage. To test this hypothesis, we examined the ability of NHFs and AT dermal fibroblasts to respond to t-butyl hydroperoxide and IR treatment. Ar fibroblasts exhibit, in comparison to NHFs, increased sensitivity to the toxicity of t-butyl hydroperoxide, as measured by colony-forming efficiency assays. Unlike NHFs, AT fibroblasts fail to show G(1) and G(2) phase checkpoint functions or to induce p53 in response to t-butyl hydroperoxide. Treatment of NHFs with t-butyl hydroperoxide activates pATM-associated kinase activity. Our results indicate that pATM is involved in responding to certain aspects of oxidative damage and in signaling this information to downstream effecters of the cell cycle checkpoint functions. Our data further suggest that some of the pathologies seen in AT could arise as a consequence of an inability to respond normally to oxidative damage. C1 NIEHS, Growth Control & Canc Grp, NIH, Res Triangle Pk, NC 27709 USA. Univ N Carolina, Sch Med, Dept Radiat Oncol, Chapel Hill, NC 27599 USA. RP Paules, RS (reprint author), NIEHS, Growth Control & Canc Grp, NIH, Mail Drop F1-05,111 Alexander Dr,POB 12233, Res Triangle Pk, NC 27709 USA. FU NCI NIH HHS [CA40453] NR 50 TC 83 Z9 83 U1 1 U2 3 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 15 PY 2001 VL 276 IS 24 BP 21951 EP 21959 DI 10.1074/jbc.M011303200 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 442RC UT WOS:000169297900150 PM 11290740 ER PT J AU Yang, FQ Zhang, TY Ito, Y AF Yang, FQ Zhang, TY Ito, Y TI Large-scale separation of resveratrol, anthraglycoside A and anthraglycoside B from Polygonum cuspidatum Sieb. et Zucc by high-speed counter-current chromatography SO JOURNAL OF CHROMATOGRAPHY A LA English DT Article DE Polygonum cuspidatum; plant materials; counter-current chromatography; resveratrol; anthraglycosides; glycosides ID PURIFICATION AB High-speed counter-current chromatography was successfully applied to the large-scale separation of resveratrol, anthraglycoside A and anthraglycoside B from the crude extract of Polygonum cuspidatum Sieb. et Zucc using a two-phase solvent system composed of chloroform, methanol and water. Resveratrol, anthraglycoside A and anthraglycoside B were separated from multigram quantities (5 g) of crude extract of P. cuspidatum. The separation yielded 200 mg to 1 g of these three compounds each at over 98% purity as determined by HPLC. The chemical structures of these components were identified by nuclear magnetic resonance (NMR) and MS. (C) 2001 Published by Elsevier Science BN. C1 NHLBI, Biophys Chem Lab, NIH, Bethesda, MD 20892 USA. Beijing Inst New Technol Applicat, Beijing Res Ctr Separat & Purificat Nat Prod, Beijing 100035, Peoples R China. RP Ito, Y (reprint author), NHLBI, Biophys Chem Lab, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 10 TC 50 Z9 55 U1 1 U2 9 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 JUN 15 PY 2001 VL 919 IS 2 BP 443 EP 448 DI 10.1016/S0021-9673(01)00846-9 PG 6 WC Biochemical Research Methods; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 442TJ UT WOS:000169300800025 PM 11442052 ER PT J AU Berger, AC Gibril, F Venzon, DJ Doppman, JL Norton, JA Bartlett, DL Libutti, SK Jensen, RT Alexander, HR AF Berger, AC Gibril, F Venzon, DJ Doppman, JL Norton, JA Bartlett, DL Libutti, SK Jensen, RT Alexander, HR TI Prognostic value of initial pasting serum gastrin levels in patients with Zollinger-Ellison syndrome SO JOURNAL OF CLINICAL ONCOLOGY LA English DT Article ID SOMATOSTATIN RECEPTOR SCINTIGRAPHY; MEDULLARY-THYROID CARCINOMA; ISLET-CELL TUMORS; DUODENAL GASTRINOMAS; LONG-TERM; LOCALIZATION; DIAGNOSIS; SECRETIN; SURGERY; CURE AB Purpose: To assess the value of the initial fasting serum gastrin (FSG) at presentation in patients with Zollinger-Ellison Syndrome (ZES) in predicting primary tumor characteristics and survival. Patients and Methods: A total of 239 patients were treated for ZES between December 1981 and September 1998, with a mean follow-up of 9.1 +/- 0.6 years. At initial evaluation, 86 patients (36%) had mild (0 to 499 pg/mL), 61 (25.5%) had moderate (500 to 1,000 pg/mL), and 92 (38.5%) had severe (> 1,000 pg/mL) elevations in FSG. Primary tumor location and size, presence of lymph node or hepatic metastases, and survival were analyzed based on the level of initial FSG, Results: In patients with sporadic ZES, but not in those with multiple endocrine neoplasia type 1 (MEN-1) and ZES, there was ct significant relationship between the level of initial FSG and tumor size and location of primary tumor, frequency of lymph node and liver metastases, and survival, The median 5- and 10-year survival decreased with increasing initial FSG (P < .001) in patients with sporadic ZES; MEN-1 patients lived longer than sporadic ZES patients (P = .012), and survival in this group was not associated with the level of initial FSG. Multivariate analysis showed that factors independently associated with death from disease in patients with sporadic ZES were liver metastases (P = .0001), a pancreatic site (P = .0027), and primary tumor size (P = .011) but not initial FSG (P > .30). Conclusion: The severity of FSG at presentation is associated with size and site of tumor and the presence of hepatic metastases, factors that are significant independent predictors of outcome. The level of FSG at presentation may be useful in planning the nature and extent of the initial evaluation and management in patients with sporadic ZES, J Clin Oncol 19:3051-3051. (C) 2001 by American Society of Clinical Oncology. C1 NCI, Surg Branch, Canc Res Ctr, NIH, Bethesda, MD 20892 USA. NCI, Data Management Sect, Canc Res Ctr, Bethesda, MD 20892 USA. NIDDKD, Digest Dis Branch, Bethesda, MD 20892 USA. NIH, Dept Diagnost Radiol, Warren G Magnuson Clin Ctr, Bethesda, MD 20892 USA. RP Alexander, HR (reprint author), NCI, Surg Branch, Canc Res Ctr, NIH, Bldg 10,Rm 2B07, Bethesda, MD 20892 USA. RI Venzon, David/B-3078-2008 NR 31 TC 16 Z9 16 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 JUN 15 PY 2001 VL 19 IS 12 BP 3051 EP 3057 PG 7 WC Oncology SC Oncology GA 442UT UT WOS:000169303900010 PM 11408501 ER PT J AU Korn, EL Arbuck, SG Pluda, JM Simon, R Kaplan, RS Christian, MC AF Korn, EL Arbuck, SG Pluda, JM Simon, R Kaplan, RS Christian, MC TI Clinical trial designs for cytostatic agents - Reply SO JOURNAL OF CLINICAL ONCOLOGY LA English DT Letter C1 NCI, Bethesda, MD 20892 USA. RP Korn, EL (reprint author), NCI, Bethesda, MD 20892 USA. NR 1 TC 4 Z9 4 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 JUN 15 PY 2001 VL 19 IS 12 BP 3154 EP 3155 PG 2 WC Oncology SC Oncology GA 442UT UT WOS:000169303900023 ER PT J AU Hoshino, T Kawase, Y Okamoto, M Yokota, K Yoshino, K Yamamura, K Miyazaki, J Young, HA Oizumi, K AF Hoshino, T Kawase, Y Okamoto, M Yokota, K Yoshino, K Yamamura, K Miyazaki, J Young, HA Oizumi, K TI Cutting edge: IL-18-transgenic mice: In vivo evidence of a broad role for IL-18 in modulating immune function SO JOURNAL OF IMMUNOLOGY LA English DT Article ID CD4(+) T-CELLS; GAMMA PRODUCTION; TH2 CELLS; EXPRESSION; CYTOKINE; INDUCTION; IGE; NK; RECEPTOR; FAMILY AB IL-18 has been shown to be a strong cofactor for Th1 T cell development. However, we previously demonstrated that when IL-18 was combined with IL-2, there was a synergistic induction of a Th2 cytokine, IL-13, in both T and NK cells. More recently, we and other groups have reported that IL-18 can potentially induce IgE, IgG1, and Th2 cytokine production in murine experimental models. Here, we report on the generation of IL-18-transgenic (Tg) mice in which mature mouse IL-18 cDNA was expressed. CD8(+)CD44(high) T cells and macrophages were increased, but B cells were decreased in these mice while serum IgE, IgG1, IL-4, and IFN-gamma levels were significantly increased. Splenic T cells in IL-18 Tg mice produced higher levels of IFN-gamma, IL-4, IL-5, and IL-13 than control wildtype mice. Thus, aberrant expression of IL-18 in vivo results in the increased production of both Th1 and Th2 cytokines. C1 Kurume Univ, Sch Med, Dept Internal Med 1, Kurume, Fukuoka 8300011, Japan. Nippon Organon KK, R&D Labs, Res & Dev Div, Osaka, Japan. Kumamoto Univ, Inst Mol Embryol Genet, Kumamoto, Japan. Osaka Univ, Sch Med, Dept Nutr & Physiol Chem, Osaka, Japan. Natl Canc Inst, Expt Immunol Lab, Frederick, MD 21702 USA. RP Hoshino, T (reprint author), Kurume Univ, Sch Med, Dept Internal Med 1, 67 Asahi Machi, Kurume, Fukuoka 8300011, Japan. RI Miyazaki, Jun-ichi/N-1976-2015 OI Miyazaki, Jun-ichi/0000-0003-2475-589X NR 24 TC 92 Z9 95 U1 0 U2 7 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD JUN 15 PY 2001 VL 166 IS 12 BP 7014 EP 7018 PG 5 WC Immunology SC Immunology GA 471VH UT WOS:000170949000002 PM 11390444 ER PT J AU Lu, J Wettstein, PJ Higashimoto, Y Appella, E Celis, E AF Lu, J Wettstein, PJ Higashimoto, Y Appella, E Celis, E TI TAP-Independent presentation of CTL epitopes by trojan antigens SO JOURNAL OF IMMUNOLOGY LA English DT Article ID MAJOR HISTOCOMPATIBILITY COMPLEX; DISTINCT PROTEOLYTIC PROCESSES; CYTOTOXIC T-CELLS; CLASS-I MOLECULES; ENDOPLASMIC-RETICULUM; ANTENNAPEDIA HOMEODOMAIN; INTRACELLULAR DELIVERY; PRESENTATION PATHWAY; SIGNAL SEQUENCE; TUMOR-ANTIGEN AB The majority of CTL epitopes are derived from intracellular proteins that are degraded in the cytoplasm by proteasomes into peptides that are transported into the endoplasmic reticulum by the TAP complex. These peptides can be further processed into the optimal size (8-10 residues) for binding with nascent MHC class I molecules, generating complexes that are exported to the cell surface. Proteins or peptides containing CTL epitopes can be introduced into the cytoplasm of APCs by linking them to membrane-translocating Trojan carriers allowing their incorporation into the MHC class I Ag-processing pathway. The present findings suggest that these "Trojan" Ags can be transported into the endoplasmic reticulum in a TAP-independent way where they are processed and trimmed into CTL epitopes. Furthermore, processing of Trojan Ags can also occur in the trans-Golgi compartment, with the participation of the endopeptidase furin and possibly with the additional participation of a carboxypeptidase. We believe that these findings will be of value for the design of CTL-inducing vaccines for the treatment or prevention of infectious and malignant diseases. C1 Mayo Clin & Mayo Grad Sch Med, Dept Immunol, Rochester, MN 55905 USA. Mayo Clin & Mayo Grad Sch Med, Ctr Canc, Rochester, MN 55905 USA. NCI, Cell Biol Lab, NIH, Bethesda, MD 20892 USA. RP Celis, E (reprint author), Mayo Clin & Mayo Grad Sch Med, Dept Immunol, GU421A, Rochester, MN 55905 USA. FU NCI NIH HHS [CA15083, CA80782, CA82677]; NCRR NIH HHS [RR00585] NR 38 TC 52 Z9 55 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 JUN 15 PY 2001 VL 166 IS 12 BP 7063 EP 7071 PG 9 WC Immunology SC Immunology GA 471VH UT WOS:000170949000008 PM 11390450 ER PT J AU Zhang, X Wang, JM Gong, WH Mukaida, N Young, HA AF Zhang, X Wang, JM Gong, WH Mukaida, N Young, HA TI Differential regulation of chemokine gene expression by 15-deoxy-Delta(12,14) prostaglandin J(2) SO JOURNAL OF IMMUNOLOGY LA English DT Article ID ACTIVATED RECEPTOR-GAMMA; NITRIC-OXIDE SYNTHASE; PPAR-GAMMA; CYCLOPENTENONE PROSTAGLANDINS; ENDOTHELIAL-CELLS; PROLIFERATOR; INTERLEUKIN-8; INHIBIT; ALPHA; TRANSCRIPTION AB Ligands for peroxisome proliferator-activated receptor gamma (PPAR gamma), such as 15-deoxy-Delta (12,14) PGJ(2) (15d-PGJ(2)) have been proposed as a new class of antiinflammatory compounds with possible clinical applications. As there is some controversy over the inhibitory effects of 15d-PGJ2 on chemokine gene expression, we investigated whether 15d-PGJ2 itself affected chemokine gene expression in human monocytes/macrophages and two monocytic cell lines. Here we demonstrate that the 15d-PGJ2 can induce IL-8 gene expression. In contrast, monocyte chemoattractant protein-1 gene expression was suppressed by 15d-PGJ2, while the expression of RANTES was unaltered. Furthermore, concomitant treatment of monocytes/macrophages with 15d-PGJ(2) (2.5 x 10(-6) M) potentiated LPS-induced gene expression of IL-8 mRNA, but suppressed PMA-induction of IL-8 mRNA. In addition, treatment of U937 and THP-1 cells with 15d-PGJ(2) also resulted in induction of IL-8 gene expression. Further studies demonstrated that 15d-PGJ2 regulated IL-8 gene expression via a ligand-specific and PPAR gamma -dependent pathway. Our observations revealed a previous unappreciated function and mechanism of 15d-PGJ(2)-mediated regulation of cytokine gene expression in monocytes/macrophages. C1 NCI, Frederick Canc Res Dev Ctr, Cellular & Mol Immunol Sect, Lab Expt Immunol,NIH, Frederick, MD 21702 USA. NCI, Frederick Canc Res Dev Ctr, Mol Immunoregulat Lab, Div Basic Sci,NIH, Frederick, MD 21702 USA. Kanazawa Univ, Canc Res Inst, Dept Mol Oncol, Kanazawa, Ishikawa 920, Japan. RP Young, HA (reprint author), NCI, Frederick Canc Res Dev Ctr, Cellular & Mol Immunol Sect, Lab Expt Immunol,NIH, Bldg 560,Rm 31-93, Frederick, MD 21702 USA. RI Zhang, Xia/B-8152-2008; Mukaida, Naofumi/D-7623-2011 OI Zhang, Xia/0000-0002-9040-1486; Mukaida, Naofumi/0000-0002-4193-1851 FU PHS HHS [N01-56000] NR 35 TC 84 Z9 88 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 JUN 15 PY 2001 VL 166 IS 12 BP 7104 EP 7111 PG 8 WC Immunology SC Immunology GA 471VH UT WOS:000170949000013 PM 11390455 ER PT J AU Zhu, JF Guo, LY Watson, CJ Hu-Li, J Paul, WE AF Zhu, JF Guo, LY Watson, CJ Hu-Li, J Paul, WE TI Stat6 is necessary and sufficient for IL-4's role in Th2 differentiation and cell expansion SO JOURNAL OF IMMUNOLOGY LA English DT Article ID INSULIN-RECEPTOR SUBSTRATE; TRANSCRIPTION FACTOR; T-CELLS; INTERLEUKIN-4 RECEPTOR; HEMATOPOIETIC-CELLS; GENE-EXPRESSION; IN-VITRO; PROTEIN; PHOSPHORYLATION; LYMPHOCYTES AB IL-4 plays a critical role in the differentiation of TCR-stimulated naive CD4 T cells to the Th2 phenotype. In response to IL-4, the IL-4R activates a set of phosphotyrosine binding domain-containing proteins, including insulin receptor substrate 1/2, Shc, and IL-4R interacting protein, as well as Stat6. Stat6 has been shown to be required for Th2 differentiation. To determine the roles of the phosphotyrosine binding adaptors in Th2 differentiation, we prepared a retrovirus containing a mutant of the human (h)IL-4R alpha -chain, Y497F, which is unable to recruit these adaptors. The mutant hIL-4R alpha, as well as the wild-type (WT) hIL-4R alpha, was introduced into naive CD4 T cells. Upon hIL-4 stimulation, Y497F worked as well as the WT hIL-4R alpha in driving Th2 differentiation, as measured by Gata3 up-regulation and IL-4 production. Furthermore, IL-4-driven cell expansion was also normal in the cells infected with Y497F, although cells infected with Y497F were not capable of phosphorylating insulin receptor substrate 2. These results suggest that the signal pathway mediated by Y497 is dispensable for both IL-4-driven Th2 differentiation and cell expansion. Both WT and Y497F hIL-4R alpha lose the ability to drive Th2 differentiation and cell expansion in Stat6-knockout CD4 T cells. A constitutively activated form of Stat6 introduced into CD4 T cells resulted in both Th2 differentiation and enhanced cell expansion. Thus, activated Stat6 is necessary and sufficient to mediate both IL-4-driven Th2 differentiation and cell expansion in CD4 T cells. C1 NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Paul, WE (reprint author), NIAID, Immunol Lab, NIH, 10 Ctr Dr MSC 1892,Bldg 10,Room 11N311, Bethesda, MD 20892 USA. RI Zhu, Jinfang/B-7574-2012 NR 42 TC 145 Z9 151 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 JUN 15 PY 2001 VL 166 IS 12 BP 7276 EP 7281 PG 6 WC Immunology SC Immunology GA 471VH UT WOS:000170949000035 PM 11390477 ER PT J AU Liu, K Li, Y Prabhu, V Young, L Becker, KG Munson, PJ Weng, NP AF Liu, K Li, Y Prabhu, V Young, L Becker, KG Munson, PJ Weng, NP TI Augmentation in expression of activation-induced genes differentiates memory from naive CD4(+) T cells and is a molecular mechanism for enhanced cellular response of memory CD4(+) T cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID HELPER CELLS; LYMPHOCYTE DEVELOPMENT; IMMUNOLOGICAL MEMORY; ACTIN CYTOSKELETON; BINDING-PROTEIN; RECEPTOR; SUBSETS; ANTIGEN; REQUIREMENTS; CLONING AB In an attempt to understand the molecular basis for the immunological memory response, we have used cDNA microarrays to measure gene expression of human memory and naive CD4(+) T cells at rest and after activation. Our analysis of 54,768 cDNA clones provides the first glimpse into gene expression patterns of memory and naive CD4(+) T cells at the genome-scale and reveals several novel findings. First, memory and naive CD4(+) T cells expressed similar numbers of genes at rest and after activation. Second, we have identified 14 cDNA clones that expressed higher levels of transcripts in memory cells than in naive cells. Third, we have identified 135 (130 known genes and 5 expressed sequence tags) up-regulated and 68 (42 known genes and 26 expressed sequence tags) down-regulated cDNA clones in memory CD4(+) T after in vitro stimulation with anti-CD3 plus anti-CD28. Interestingly, the increase in mRNA levels of up-regulated genes was greater in memory than in naive CD4(+) T cells after in vitro stimulation and was higher with anti-CD3 plus anti-CD28 than with anti-CD3 alone in both memory and naive CD4(+) T cells. Finally, the changes in expression of actin and cytokine genes identified by cDNA microarrays were confirmed by Northern and protein analyses. Together, we have identified similar to 200 cDNA clones whose expression levels changed after activation and suggest that the level of expression of up-regulated genes is a molecular mechanism that differentiates the response of memory from naive CD4(+) T cells. C1 NIA, Immunol Lab, NIH, Baltimore, MD 21224 USA. NIH, Math & Stat Comp Lab, Ctr Informat Technol, Bethesda, MD 20892 USA. NIA, DNA Array Unit, NIH, Baltimore, MD 21224 USA. RP Weng, NP (reprint author), NIA, Immunol Lab, NIH, 5600 Nathan Shock Dr,Box 21, Baltimore, MD 21224 USA. OI Liu, Kebin/0000-0003-1965-7240; Becker, Kevin/0000-0002-6794-6656 NR 67 TC 48 Z9 50 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 JUN 15 PY 2001 VL 166 IS 12 BP 7335 EP 7344 PG 10 WC Immunology SC Immunology GA 471VH UT WOS:000170949000042 PM 11390484 ER PT J AU Benton, KA Misplon, JA Lo, CY Brutkiewicz, RR Prasad, SA Epstein, SL AF Benton, KA Misplon, JA Lo, CY Brutkiewicz, RR Prasad, SA Epstein, SL TI Heterosubtypic immunity to influenza A virus in mice lacking IgA, all Ig, NKT cells, or gamma delta T cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID A VIRUS; MONOCLONAL-ANTIBODIES; DEFICIENT MICE; B-CELLS; HETEROTYPIC IMMUNITY; NUCLEOPROTEIN GENE; RESPIRATORY-TRACT; MURINE INFLUENZA; LYMPHOCYTES-T; MUTANT MICE AB The mechanisms of broad cross-protection to influenza viruses of different subtypes, termed heterosubtypic immunity, remain incompletely understood. We used knockout mouse strains to examine the potential for heterosubtypic immunity in mice lacking IgA, all Ig and B cells, NKT cells (CD1 knockout mice), or gamma delta T cells. Mice were immunized with live influenza A virus and compared with controls immunized with unrelated influenza B virus. IgA(-/-) mice survived full respiratory tract challenge with heterosubtypic virus that was lethal to controls. IgA(-/-) mice also cleared virus from the nasopharynx and lungs following heterosubtypic challenge limited to the upper respiratory tract, where IgA has been shown to play an important role. Ig(-/-) mice controlled the replication of heterosubtypic challenge virus in the lungs. Acute depletion of CD4(+) or CD8(+) T cell subsets abrogated this clearance of virus, thus indicating that both CD4(+) and CD8(+) T cells are required for protection in the absence of Ig. These results in Ig(-/-) mice indicate that CD4(+) T cells can function by mechanisms other than providing help to B cells for the generation of Abs. Like wild-type mice, CD1(-/-) mice and gamma delta (-/-) mice survived lethal heterosubtypic challenge. Acute depletion of CD4(+) and CD8(+) cells abrogated heterosubtypic protection in gamma delta (-/-) mice, but not B6 controls, suggesting a contribution of gamma delta T cells. Our results demonstrate that the Ab and cellular subsets deficient in these knockout mice are not required for heterosubtypic protection, but each may play a role in a multifaceted response that as a whole is more effective than any of its parts. C1 US FDA, Ctr Biol Evaluat & Res, Off Therapeut Res & Review, Div Cellular & Gene Therapies,Mol Immunol Lab, Bethesda, MD 20852 USA. NIAID, Viral Dis Lab, Cellular Biol Sect, NIH, Bethesda, MD 20892 USA. NIAID, Viral Immunol Sect, NIH, Bethesda, MD 20892 USA. Indiana Univ, Sch Med, Dept Microbiol & Immunol, Walther Oncol Ctr, Indianapolis, IN 46202 USA. RP Benton, KA (reprint author), US FDA, Ctr Biol Evaluat & Res, Off Therapeut Res & Review, Div Cellular & Gene Therapies,Mol Immunol Lab, 1401 Rockville Pike,HFM-521, Bethesda, MD 20852 USA. NR 72 TC 97 Z9 103 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 JUN 15 PY 2001 VL 166 IS 12 BP 7437 EP 7445 PG 9 WC Immunology SC Immunology GA 471VH UT WOS:000170949000054 PM 11390496 ER PT J AU Salcedo, R Young, HA Ponce, ML Ward, JM Kleinman, HK Murphy, WJ Oppenheim, JJ AF Salcedo, R Young, HA Ponce, ML Ward, JM Kleinman, HK Murphy, WJ Oppenheim, JJ TI Eotaxin (CCL11) induces in vivo angiogenic responses by human CCR3(+) endothelial cells SO JOURNAL OF IMMUNOLOGY LA English DT Article ID CHEMOKINE RECEPTORS; IFN-GAMMA; IN-VIVO; CXC CHEMOKINES; EXPRESSION; EOSINOPHILS; ALPHA; GROWTH; BRAIN; ANTIANGIOGENESIS AB Chemokines are attractants and regulators of cell activation. Several CXC family chemokine members induce angiogenesis and promote tumor growth. In contrast, the only CC chemokine, reported to play a direct role in angiogenesis is monocyte-chemotactic protein-1. Here we report that another CC chemokine, eotaxin (also known as CCL11), also induced chemotaxis of human microvascular endothelial cells. CCL11-induced chemotactic responses were comparable with those induced by monocyte-chemotactic protein-1 (CCL2), but lower than those induced by stroma-derived factor-la (CXCL12) and IL-8 (CXCL8). The chemotactic activity was consistent with the expression of CCR3, the receptor for CCL11, on human microvascular endothelial cells and was inhibited by mAbs to either human CCL11 or human CCR3. CCL11 also induced the formation of blood vessels in vivo as assessed by the chick chorioallantoic membrane and Matrigel plug assays. The angiogenic response induced by CCL11 was about one-half of that induced by basic fibroblast factor, and it was accompanied by an inflammatory infiltrate, which consisted predominantly of eosinophils. Because the rat aortic sprouting assay, which is not infiltrated by eosinophils, yielded a positive response to CCL11, this angiogenic response appears to be direct and is not mediated by eosinophil products. This suggests that CCL11 may contribute to angiogenesis in conditions characterized by increased CCL11 production and eosinophil infiltration such as Hodgkin's lymphoma, nasal polyposis, endometriosis, and allergic diathesis. C1 NCI, Frederick Canc Res & Dev Ctr, Mol Immunoregulat Lab, Frederick, MD 21702 USA. Frederick Canc Res & Dev Ctr, Expt Immunol Lab, Div Basic Sci, Frederick, MD 21702 USA. Frederick Canc Res & Dev Ctr, Sci Applicat Int Corp, Intramural Res Support Program, Frederick, MD 21702 USA. Frederick Canc Res & Dev Ctr, Off Lab Anim Resources, Vet & Tumor Pathol Sect, Frederick, MD 21702 USA. Natl Inst Dent & Craniofacial Res, Cell Biol Sect, Bethesda, MD 20892 USA. RP Salcedo, R (reprint author), NCI, Frederick Canc Res & Dev Ctr, Mol Immunoregulat Lab, Bldg 567,Room 209, Frederick, MD 21702 USA. FU NCI NIH HHS [N01-CO-56000] NR 44 TC 132 Z9 137 U1 0 U2 8 PU AMER ASSOC IMMUNOLOGISTS PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0022-1767 J9 J IMMUNOL JI J. Immunol. PD JUN 15 PY 2001 VL 166 IS 12 BP 7571 EP 7578 PG 8 WC Immunology SC Immunology GA 471VH UT WOS:000170949000071 PM 11390513 ER PT J AU Milne, E Grafman, J AF Milne, E Grafman, J TI Ventromedial prefrontal cortex lesions in humans eliminate implicit gender stereotyping SO JOURNAL OF NEUROSCIENCE LA English DT Article DE prefrontal cortex; social cognition; stereotypes; implicit association; social attitudes; ventromedial cortex ID COGNITION; ASSOCIATION; ACTIVATION; PATIENT; DAMAGE; MEMORY; GAUGE AB Patients with prefrontal cortex lesions and controls were administered an implicit association task (IAT) that measured the degree of association between male and female names and their stereotypical attributes of strength and weakness. They also completed three questionnaires measuring their explicit judgment regarding gender-related stereotypical attributes. There were no between-group differences on the explicit measures. On the IAT, patients with dorsolateral lesions and controls showed a strong association, whereas patients with ventromedial prefrontal cortex lesions had a significantly lower association, between the stereotypical attributes of men and women and their concepts of gender. This finding provides support for the hypothesis that patients with ventromedial prefrontal lesions have a deficit in automatically accessing certain aspects of overlearned associated social knowledge. C1 NINDS, Cognit Neurosci Sect, NIH, Bethesda, MD 20892 USA. RP Grafman, J (reprint author), NINDS, Cognit Neurosci Sect, NIH, Bldg 10,Rm 5C205,MSC 1440, Bethesda, MD 20892 USA. NR 29 TC 35 Z9 36 U1 0 U2 12 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 JUN 15 PY 2001 VL 21 IS 12 BP art. no. EP RC150 PG 6 WC Neurosciences SC Neurosciences & Neurology GA 442HV UT WOS:000169279900001 PM 11404442 ER PT J AU Kaczmarczyk, SJ Green, JE AF Kaczmarczyk, Stanislaw J. Green, Jeffrey E. TI A single vector containing modified cre recombinase and LOX recombination sequences for inducible tissue-specific amplification of gene expression SO NUCLEIC ACIDS RESEARCH LA English DT Article AB The selective alteration of the genome using Cre recombinase to target the rearrangement of genes flanked by LOX recognition sequences has required the use of two separate genetic constructs in trans, one containing cre and the other containing the gene of interest flanked by LOX sites. We have developed a strategy in which both the cre recombinase gene and LOX recombination sites may be cloned within a single vector in cis. This method uses a modified form of Cre (CREM) that contains alterations to the 5' region including the introduction of a Kozak consensus sequence and insertion of a functional intron. This system allows for the inducible, tissue-specific activation or inactivation of gene expression in a single vector and can be utilized for the 300-fold amplification of gene expression from a weak promoter. This approach can be applied to targeting strategies for generating genetically altered mice and gene therapy. C1 [Kaczmarczyk, Stanislaw J.; Green, Jeffrey E.] NCI, Transgen Oncogenesis Grp, Lab Cell Regulat & Carcinogenesis, Bethesda, MD 20892 USA. RP Green, JE (reprint author), NCI, Transgen Oncogenesis Grp, Lab Cell Regulat & Carcinogenesis, Bethesda, MD 20892 USA. EM jegreen@nih.gov NR 31 TC 33 Z9 34 U1 0 U2 1 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0305-1048 J9 NUCLEIC ACIDS RES JI Nucleic Acids Res. PD JUN 15 PY 2001 VL 29 IS 12 AR e56 DI 10.1093/nar/29.12.e56 PG 13 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA V17WT UT WOS:000207968000002 PM 11410679 ER PT J AU Augenstein, KF McVeigh, ER Young, AA AF Augenstein, KF McVeigh, ER Young, AA TI Magnetic resonance imaging and ventricle mechanics SO PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY A-MATHEMATICAL PHYSICAL AND ENGINEERING SCIENCES LA English DT Article DE magnetic resonance imaging; cardiac mechanics; image processing ID TAGGED MR-IMAGES; DILATED CARDIOMYOPATHY; HYPERTROPHIC CARDIOMYOPATHY; MYOCARDIAL STRAIN; CONDUCTION DELAY; PACED HEART; DEFORMATION; ACTIVATION; SEQUENCE; CHAMBER AB Methods for cardiac geometry and motion reconstruction from multi-planar tagged magnetic resonance images (MRI) have been developed and refined over the last decade. This paper firstly provides an overview of a finite-element reconstruction method, including variants for interactive three-dimensional reconstruction. The second part of this paper discusses recent applications of MRI and ventricular modelling in the study of electrical-mechanical coupling. C1 Univ Auckland, Dept Engn Sci, Auckland, New Zealand. Univ Auckland, Dept Anat, Auckland, New Zealand. Univ Auckland, Dept Physiol, Auckland, New Zealand. Johns Hopkins Univ, Sch Med, Cardiac Energet Lab, NHLBI, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Dept Biomed Engn, Baltimore, MD 21205 USA. RP Augenstein, KF (reprint author), Univ Auckland, Dept Engn Sci, Private Bag 92019, Auckland, New Zealand. OI Young, Alistair/0000-0001-5702-4220 NR 42 TC 4 Z9 4 U1 1 U2 2 PU ROYAL SOC PI LONDON PA 6-9 CARLTON HOUSE TERRACE, LONDON SW1Y 5AG, ENGLAND SN 1364-503X EI 1471-2962 J9 PHILOS T R SOC A JI Philos. Trans. R. Soc. A-Math. Phys. Eng. Sci. PD JUN 15 PY 2001 VL 359 IS 1783 BP 1263 EP 1275 DI 10.1098/rsta.2001.0830 PG 13 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 443TM UT WOS:000169356800016 ER PT J AU Davies, C Tingley, D Kachar, B Wenthold, RJ Petralia, RS AF Davies, C Tingley, D Kachar, B Wenthold, RJ Petralia, RS TI Distribution of members of the PSD-95 family of MAGUK proteins at the synaptic region of inner and outer hair cells of the guinea pig cochlea SO SYNAPSE LA English DT Article DE PSD-93; SAP-97; SAP-102; synapse ID GLUTAMATE-RECEPTOR SUBUNIT; TUMOR-SUPPRESSOR PROTEIN; EFFERENT NERVE-TERMINALS; NITRIC-OXIDE SYNTHASE; K+ CHANNEL; POSTSYNAPTIC DENSITY-93; CLUSTERING ACTIVITY; GUANYLATE KINASES; AMPA RECEPTORS; NMDA RECEPTORS AB PDZ-domain containing proteins of the MAGUK (membrane-associated guanylate kinase) family target, anchor, and cluster receptors and channels to subcellular sites. Among the MAGUK proteins, the members of the PSD-95 family (MAGUKs: PSD-95, PSD-93, SAP-97, and SAP-102) target and anchor glutamate receptors to the synaptic terminals. Associations of glutamate receptors with MAGUKs have been described in the brain but not in the cochlea. In this study, RT-PCR, immunofluorescence microscopy, and immunoelectron microscopy were used to investigate the presence and distribution of MAGUK proteins in the organ of Corti. The presence of the mRNA for PSD-95, PSD-93, SAP-97, and SAP-102 in the organ of Corti was confirmed by RT-PCR, Immunocytochemistry using a "pan-MAGUK" antibody, which recognizes all four MAGUK proteins, and selective antibodies against these proteins revealed that all four MAGUKs are present within the base of inner hair cells while all except SAP-97 are found within the base of the outer hair cells. In addition, PSD-93 and PSD-95 are found in postsynaptic afferent terminals on inner hair cells, while postsynaptic afferent terminals on outer hair cells have PSD-93. Synapse 40:258-268, 2001, (C) 2001 Wiley-Liss, Inc.dagger C1 NIDCD, Neurochem Lab, NIH, Bethesda, MD 20892 USA. NIDCD, Sect Struct Cell Biol, NIH, Bethesda, MD USA. RP Petralia, RS (reprint author), NIDCD, Neurochem Lab, NIH, Bldg 36,Room 5D08, Bethesda, MD 20892 USA. NR 52 TC 26 Z9 26 U1 0 U2 4 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0887-4476 J9 SYNAPSE JI Synapse PD JUN 15 PY 2001 VL 40 IS 4 BP 258 EP 268 DI 10.1002/syn.1048 PG 11 WC Neurosciences SC Neurosciences & Neurology GA 428LV UT WOS:000168463900002 PM 11309841 ER PT J AU Xu, H Lu, YF Thomas, JB Carroll, FI Rice, KC Rothman, RB AF Xu, H Lu, YF Thomas, JB Carroll, FI Rice, KC Rothman, RB TI Opioid peptide receptor studies. 15. Relative efficacy of 4-[(N-allyl-3-methyl-4 piperidinyl) phenylamino]-N,N-diethylbenzamide and related compounds at the cloned human delta-opioid receptor SO SYNAPSE LA English DT Article DE opioid receptors; analgesics; [S-35]GTP-gamma-S binding; intrinsic efficacy ID SELECTIVE LIGANDS; AGONIST EFFICACY; BINDING-SITES; INHIBITION; BIOASSAY; TAN-67; BRAIN; DRUGS AB Previous data obtained from both binding and functional assays demonstrated that (-)-4-[(N-allyl-3-methyl-4-piperidinyl)phenylamino]-N,N-diethylbenzamide [(-)-RTI5989-54] displays selective binding and full agonist activity relative to (+/-)-RTI5989-54 for the delta opioid receptor. The present study was conducted to evaluate the activities of structurally diverse opioid receptor delta ligands in the [S-35]GTP-gamma -S binding assay, comparing the relationship between receptor binding, activation, efficacy, and intrinsic efficacy. The data, obtained with cloned human delta receptors, demonstrated that (-)-RTI5989-54 behaves like the highly selective delta agonist SNC80. Addition of the hydroxyl group to RTI5989-54 (RTI5989-61) or replacement of the allyl group with the trans-crotyl group on the piperidine nitrogen of RTI-5989-61 (RTI5989-62) increased binding affinity, produced full agonist activity, and decreased intrinsic efficacy at the delta opioid receptor. The order of potency for the EC50 (GTP-gamma -S) was RTI5989-62 (0.20 nM) > RTI5989-61 (0.43 nM) > SNC80 (1.92 nM) > DPDPE (3.50 nM) (-)-RTI5989-54 (17.6 nM) > (+/-)-RTI5989-54 (65.6 nM) > (+)-RTI5989-54 (483 nM). RTI5989-61 and RTI5989-62 were fully efficacious, but had intrinsic efficacy values that were 2.2-3.1 times lower than that of DPDPE and SNC80. Comparison of the binding K-i in competitively inhibiting [I-125]IOXY binding to the functional K-i for delta antagonists [Ki (IOXY)/Ki (GTP-gamma -S)] shows that antagonists might antagonize agonist-evoked neurochemical effects with equal magnitude while occupying different proportions of target receptors. Synapse 40:289-274, 2001, (C) 2001 Wiley-Liss, Inc.dagger C1 NIDA, CPS, IRP, DIR,NIH, Baltimore, MD 21224 USA. Res Triangle Inst, Res Triangle Pk, NC 27709 USA. NIDDK, LMC, NIH, Bethesda, MD USA. RP Rothman, RB (reprint author), NIDA, CPS, IRP, DIR,NIH, POB 5180,5500 Nathan Shock Dr, Baltimore, MD 21224 USA. FU NIDA NIH HHS [DA09045] NR 24 TC 4 Z9 4 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 0887-4476 J9 SYNAPSE JI Synapse PD JUN 15 PY 2001 VL 40 IS 4 BP 269 EP 274 DI 10.1002/syn.1049 PG 6 WC Neurosciences SC Neurosciences & Neurology GA 428LV UT WOS:000168463900003 PM 11309842 ER PT J AU Vinade, L Petersen, JD Do, K Dosemeci, A Reese, TS AF Vinade, L Petersen, JD Do, K Dosemeci, A Reese, TS TI Activation of calpain may alter the postsynaptic density structure and modulate anchoring of NMDA receptors SO SYNAPSE LA English DT Article DE protease; NMDA receptor; PSD-95; anchoring proteins; rotary shadowing ID TRANSIENT CEREBRAL-ISCHEMIA; ENTORHINAL CORTEX PRODUCE; RAT-BRAIN; MEDIATED DEGRADATION; EXCITATORY SYNAPSES; GUANYLATE KINASES; PROTEIN PSD-95; DENTATE GYRUS; FAMILY; CALCIUM AB Elevation of calcium during sustained synaptic activity may lead to the activation of the postsynaptic calcium-dependent protease calpain and thus could alter the integrity and localization of endogenous proteins. The distribution of anchoring proteins for neuroreceptors is an important determinant of the efficacy of neuronal transmission. Many of these anchoring proteins are concentrated within the postsynaptic density (PSD). In the present study, we examined the effects of calpain II on isolated PSDs using biochemical and electron microscopic techniques. Biochemical analysis reveals that PSD-95, a clustering molecule which anchors NMDA receptors by interaction with their NR2 subunits, as well as the NR2 subunits themselves, are cleaved by calpain. On the other hand, under conditions where all the PSD-95 protein is cleaved, actin and alpha -actinin-a protein thought to anchor NMDA receptors to actin filaments-remain intact. For analysis by electron microscopy, PSDs were adsorbed on glass, immunogold-labeled with an antibody to PSD-95, slam frozen, freeze dried, and rotary shadowed. Electron micrographs of replicas indicate that PSDs are disc-shaped and are composed of a lattice-like structure which labels with PSD-95 immunogold. After calpain treatment, PSDs adsorbed on glass become thinner overall and the lattice becomes fragmented. Altogether, these results suggest that calpain activity could produce changes in the organization of the PSD and, by cleaving PSD-95 associated with the PSD lattice, could modify the anchoring of NMDA receptors. Synapse 40:302-309, 2001. (C) 2001 Wiley-Liss, Inc.dagger C1 NINDS, Neurobiol Lab, NIH, Bethesda, MD 20892 USA. RP Vinade, L (reprint author), NINDS, Neurobiol Lab, NIH, Bldg 36-2A21, Bethesda, MD 20892 USA. OI Petersen, Jennifer/0000-0003-1107-8535 NR 51 TC 22 Z9 23 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 0887-4476 J9 SYNAPSE JI Synapse PD JUN 15 PY 2001 VL 40 IS 4 BP 302 EP 309 DI 10.1002/syn.1053 PG 8 WC Neurosciences SC Neurosciences & Neurology GA 428LV UT WOS:000168463900007 PM 11309846 ER PT J AU Allison, TJ Winter, CC Fournie, JJ Bonneville, M Garboczi, DN AF Allison, TJ Winter, CC Fournie, JJ Bonneville, M Garboczi, DN TI Structure of a human gamma delta T-cell antigen receptor SO NATURE LA English DT Article ID NONPEPTIDIC MYCOBACTERIAL LIGANDS; RECOGNITION; STIMULATION; CRYSTALLOGRAPHY; PYROPHOSPHATE; SUBSET; SUITE AB T-cell antigen receptors composed of gamma and delta polypeptide chains (gamma delta TCRs) can directly recognize antigens in the form of intact proteins or non-peptide compounds, unlike alpha beta TCRs, which recognize antigens bound to major histocompatibility complex molecules (MHC). About 5% of peripheral blood T cells bear gamma delta TCRs, most of which recognize non-peptide phosphorylated antigens(1,2). Here we describe the 3.1 Angstrom resolution structure of a human gamma delta TCR from a T-cell clone(3) that is phosphoantigen-reactive. The orientation of the variable (V) and constant (C) regions of the gamma delta TCR is unique when compared with alpha beta TCRs or antibodies, and results from an unusually small angle between the V gamma and C gamma domains. The complementarity-determining regions (CDRs) of the V domains exhibit a chemically reasonable binding site for phosphorylated antigens, providing a possible explanation for the canonical usage of the V gamma9 and V delta2 gene segments by phosphoantigen-reactive receptors. Although the gamma delta TCR V domains are similar in overall structure to those of alpha beta TCRs, gamma delta TCR C domains are markedly different. Structural differences in C gamma and C delta, and in the location of the disulphide bond between them, may enable gamma delta TCRs to form different recognition/signalling complexes than alpha beta TCRs. C1 NIAID, Struct Biol Sect, Immunogenet Lab, Rockville, MD 20852 USA. CHU Purpan, INSERM, U395, F-31024 Toulouse, France. Inst Biol, INSERM, U463, F-44035 Nantes, France. RP Allison, TJ (reprint author), NIAID, Struct Biol Sect, Immunogenet Lab, 12441 Parklawn Dr, Rockville, MD 20852 USA. RI Fournie, jean-jacques/J-7805-2013 OI Fournie, jean-jacques/0000-0001-6542-6908 NR 30 TC 152 Z9 156 U1 3 U2 7 PU MACMILLAN PUBLISHERS LTD PI LONDON PA PORTERS SOUTH, 4 CRINAN ST, LONDON N1 9XW, ENGLAND SN 0028-0836 J9 NATURE JI Nature PD JUN 14 PY 2001 VL 411 IS 6839 BP 820 EP 824 DI 10.1038/35081115 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 441TV UT WOS:000169246400055 PM 11459064 ER PT J AU Gollob, MH Green, MS Tang, ASL Gollob, T Karibe, A Roberts, R Ahmad, F Lozado, R Shah, G Fananapazir, L Bachinski, LL Roberts, R Tapscott, T Gonzales, O Begley, D Mohiddin, S AF Gollob, MH Green, MS Tang, ASL Gollob, T Karibe, A Roberts, R Ahmad, F Lozado, R Shah, G Fananapazir, L Bachinski, LL Roberts, R Tapscott, T Gonzales, O Begley, D Mohiddin, S TI Identification of a gene responsible for familial Wolff-Parkinson-White syndrome SO NEW ENGLAND JOURNAL OF MEDICINE LA English DT Article; Proceedings Paper CT 22nd Annual Meeting of the North-American-Society-of-Pacing-and-Electrophysiology CY MAY 02-05, 2001 CL BOSTON, MA SP N Amer Soc Pacing & Electrophysiol ID ACTIVATED PROTEIN-KINASE; ATRIAL-FIBRILLATION; CARDIAC-ARREST; PATTERNS; SUBUNIT; YOUNG AB Background The Wolff Parkinson White syndrome, with a prevalence in Western countries of 1.5 to 3.1 per 1000 persons, causes considerable morbidity and may cause sudden death. We identified two families in which the Wolff Parkinson White syndrome segregated as an autosomal dominant disorder. Methods We studied 70 members of the two families (57 in Family 1 and 13 in Family 2). The subjects underwent 12-lead electrocardiography and two-dimensional echocardiography. Genotyping mapped the gene responsible to 7q34-q36, a locus previously identified to be responsible for an inherited form of Wolff Parkinson White syndrome. Candidate genes were identified, sequenced, and analyzed in normal and affected family members to identify the disease-causing gene. Results A total of 31 members (23 from Family 1 and 8 from Family 2) had the Wolff Parkinson White syndrome. Affected members of both families had ventricular preexcitation with conduction abnormalities and cardiac hypertrophy. The maximal combined two-point lod score was 9.82 at a distance of 5 cM from marker D7S636, which confirmed the linkage of the gene in both families to 7q34-q36. Haplotype analysis indicated that there were no alleles in common in the two families at this locus, suggesting that the two families do not have a common founder. We identified a missense mutation in the gene that encodes the gamma2 regulatory subunit of AMP-activated protein kinase (PRKAG2). The mutation results in the substitution of glutamine for arginine at residue 302 in the protein. Conclusions The identification of this genetic defect has important implications for elucidating the pathogenesis of ventricular preexcitation. Further understanding of how this molecular defect leads to supraventricular arrhythmias could influence the development of specific therapies for other forms of supraventricular arrhythmia. (N Engl J Med 2001;344:1823-31.) Copyright (C) 2001 Massachusetts Medical Society. C1 Baylor Coll Med, Dept Med, Cardiol Sect, Houston, TX 77030 USA. Univ Ottawa, Inst Heart, Ottawa, ON, Canada. NHLBI, Inherited Heart Dis Sect, NIH, Bethesda, MD 20892 USA. RP Roberts, R (reprint author), Baylor Coll Med, Dept Med, Cardiol Sect, 6550 Fannin,MS SM 677, Houston, TX 77030 USA. EM rroberts@bcm.tmc.edu RI Tang, Anthony/E-6203-2014 NR 28 TC 317 Z9 338 U1 0 U2 7 PU MASSACHUSETTS MEDICAL SOC PI WALTHAM PA WALTHAM WOODS CENTER, 860 WINTER ST,, WALTHAM, MA 02451-1413 USA SN 0028-4793 J9 NEW ENGL J MED JI N. Engl. J. Med. PD JUN 14 PY 2001 VL 344 IS 24 BP 1823 EP 1831 DI 10.1056/NEJM200106143442403 PG 9 WC Medicine, General & Internal SC General & Internal Medicine GA 441LC UT WOS:000169231000003 PM 11407343 ER PT J AU Maurici, D Monti, P Campomenosi, P North, S Frebourg, T Fronza, G Hainaut, P AF Maurici, D Monti, P Campomenosi, P North, S Frebourg, T Fronza, G Hainaut, P TI Amifostine (WR2721) restores transcriptional activity of specific p53 mutant proteins in a yeast functional assay SO ONCOGENE LA English DT Article DE amifostine; p53; yeast functional assay ID TUMOR-SUPPRESSOR P53; HUMAN-MELANOMA CELLS; DNA-BINDING; IN-VITRO; ESOPHAGEAL CANCERS; GENE-EXPRESSION; CYCLE ARREST; BAX GENE; APOPTOSIS; MUTATIONS AB Many p53 mutants found in human cancer have an altered ability to bind DNA and transactivate gene expression. Re-expression of functional p53 in cells in which the endogenous TP53 gene is inactivated has been demonstrated to restore a non-tumorigenic phenotype, Pharmacological modulation of p53 mutant conformation may therefore represent a mechanism to reactivate p53 function and consequently improve response to radio- and chemotherapy, We have recently reported that the radio- and chemoprotector Amifostine (WR2721, Ethyol(H)) activates wild-type p53 in cultured mammalian cells. In the present study, we have used a yeast functional assay to investigate the effect of WR2721 on the transcriptional activity of p53. WR2721 restored this activity in a temperature-sensitive mutant V272M (valine to methionine at codon 272) expressed at the nonpermissive temperature and it also partially restored the transcriptional activity of several other conformationally flexible p53 mutants. The results indicate that the yeast functional assay may be used to identify compounds that modulate p53 activity, with potential therapeutic implications. C1 Int Agcy Res Canc, Grp Mol Carcinogenesis, F-69372 Lyon, France. Natl Canc Inst, Mutagenesis Lab, IST, I-16132 Genoa, Italy. Fac Med & Pharm, INSERM EPI, F-76183 Rouen, France. RP Hainaut, P (reprint author), Int Agcy Res Canc, Grp Mol Carcinogenesis, 150 Cours Albert Thomas, F-69372 Lyon, France. RI Hainaut, Pierre /B-6018-2012; Campomenosi, Paola/C-9729-2011; OI Hainaut, Pierre /0000-0002-1303-1610; Campomenosi, Paola/0000-0002-8853-1134 NR 44 TC 41 Z9 43 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD JUN 14 PY 2001 VL 20 IS 27 BP 3533 EP 3540 DI 10.1038/sj.onc.1204428 PG 8 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 445VB UT WOS:000169478300009 PM 11429700 ER PT J AU Campomenosi, P Monti, P Aprile, A Abbondandolo, A Frebourg, T Gold, B Crook, T Inga, A Resnick, MA Iggo, R Fronza, G AF Campomenosi, P Monti, P Aprile, A Abbondandolo, A Frebourg, T Gold, B Crook, T Inga, A Resnick, MA Iggo, R Fronza, G TI p53 mutants can often transactivate promoters containing a p21 but not Bax or PIG3 responsive elements SO ONCOGENE LA English DT Article DE p53; yeast functional assay; p53 responsive elements ID YEAST FUNCTIONAL ASSAY; CELL-CYCLE ARREST; DNA-BINDING; BREAST-CANCER; MUTATIONS; PROTEIN; APOPTOSIS; TUMORS; TRANSFORMATION; IDENTIFICATION AB The human p53 protein acts mainly as a stress inducible transcription factor transactivating several genes involved in cell cycle arrest (e,g, p21) or apoptosis (e,g, Bax, PIG3), Roughly half of all human tumours contains p53 missense mutations. Virtually all tumour-derived p53 mutants are unable to activate Bax transcription but some retain the ability to activate p21 transcription, Identification of these mutants may have valuable clinical implications. We have determined the transactivation ability of 77 p53 mutants using reporter yeast strains containing a p53-regulated ADE2 gene whose promoter is regulated by p53 responsive elements derived from the regulatory region of the p21, BRS and PIG3 genes, We also assessed the influence of temperature on transactivation, Our results indicate that a significant proportion of mutants [16/77 (21%); 10/64 (16%) considering only tumour-derived mutants] are transcriptionally active, especially with the p21 promoter. Discriminant mutants preferentially affect less conserved (P < 0.04, Fisher's exact test), more rarely mutated (P < 0.006, Fisher's exact test) amino acids. Temperature sensitivity is frequently observed, but is more common among discriminant than non-discriminant mutants (P < 0.003, Fisher's exact test), Finally, we extended the analysis to a group of mutants isolated in BRCA-associated tumours that surprisingly were indistinguishable from wild type in standard transcription, growth suppression and apoptosis assays in human cells, but showed gain of function in transformation assays. The incidence of transcriptionally active mutations among this group was significantly higher than in the panel of mutants studied previously (P < 0.001, Fisher's exact test), Since it is not possible to predict the behaviour of a mutant from first principles, we propose that the yeast assay be used to compile a functional p53 database and fill the gap between the biophysical, pharmacological and clinical fields. C1 Natl Inst Canc Res, Mutagenesis Lab, I-16132 Genoa, Italy. Univ Genoa, Dept Oncol Biol & Genet, Genoa, Italy. IFRMP, INSERM EP19906, Fac Med & Pharm, F-76183 Rouen, France. Univ Nebraska, Med Ctr, Dept Pharmaceut Sci, Omaha, NE 68198 USA. St Marys Hosp, Sch Med, Ludwig Inst Canc Res, Sect Cell Biol & Expt Pathol, London W2 1PG, England. NIEHS, Chromosome Stabil Grp, NIH, Res Triangle Pk, NC 27709 USA. Swiss Inst Expt Canc Res, ISREC, CH-1066 Epalinges, Switzerland. RP Fronza, G (reprint author), Natl Inst Canc Res, Mutagenesis Lab, Largo R Benzi 10, I-16132 Genoa, Italy. RI Iggo, Richard/G-3546-2014; Campomenosi, Paola/C-9729-2011; OI Campomenosi, Paola/0000-0002-8853-1134 FU NCI NIH HHS [R01 CA29088] NR 38 TC 96 Z9 99 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD JUN 14 PY 2001 VL 20 IS 27 BP 3573 EP 3579 DI 10.1038/sj.onc.1204468 PG 7 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 445VB UT WOS:000169478300014 PM 11429705 ER PT J AU Tao, T Skiadopoulos, MH Davoodi, F Surman, SR Collins, PL Murphy, BR AF Tao, T Skiadopoulos, MH Davoodi, F Surman, SR Collins, PL Murphy, BR TI Construction of a live-attenuated bivalent vaccine virus against human parainfluenza virus (PIV) types 1 and 2 using a recombinant PIV3 backbone SO VACCINE LA English DT Article DE vaccine; human parainfluenza virus (PIV); recombinant PIV3 backbone ID RESPIRATORY SYNCYTIAL VIRUS; VESICULAR STOMATITIS VIRUSES; TEMPERATURE-SENSITIVE MUTANTS; SENDAI-VIRUS; FOREIGN GENE; HEMAGGLUTININ-NEURAMINIDASE; CANDIDATE VACCINE; MEASLES-VIRUS; FUSION GLYCOPROTEINS; STABLE EXPRESSION AB PIV1 and PIV2 are important agents of pediatric respiratory tract disease. We are developing live-attenuated vaccines against these viruses. We earlier constructed a PIV3/PIV1 antigenic chimeric virus, designated rPIV3-1, in which the hemagglutinin-neuraminidase (HN) and fusion (F) proteins of wild type rPIV3 were replaced by their PIV1 counterparts. In the present study, rPIV3-1 was used as a vector to express the HN protein of PIV2, to generate a single virus capable of inducing immunity to both PIV1 and PIV2. The PIV2 HN open reading frame was expressed from an extra gene cassette, under the control of PIV3 cis-acting transcription signals, inserted between the F and HN genes of rPIV3-1. The recombinant derivative, designated rPIV3-1.2HN, was readily recovered and exhibited a level of temperature sensitivity and in vitro growth similar to that of its parental virus. The rPIV3-1.2HN virus was restricted in replication in both the upper and lower respiratory tracts of hamsters compared with rPIV3-1, identifying an attenuating effect of the PIV2 HN insert in hamsters. rPIV3-1.2HN elicited serum antibodies to both PIV1 and PIV2 and induced resistance against challenge with wild type PIV1 or PIV2. Thus, rPIV3-1.2HN, a virus attenuated solely by the insertion of the PIV2 HN gene, functioned as a live attenuated bivalent vaccine candidate against both PIV1 and PIV2. Published by Elsevier Science Ltd. C1 NIAID, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. RP Murphy, BR (reprint author), NIAID, Infect Dis Lab, NIH, Bldg 7,Rm 106,7 Ctr Dr,MSC 0720, Bethesda, MD 20892 USA. EM bm25f@nih.gov FU NIAID NIH HHS [AI-000099, AI-000087] NR 59 TC 10 Z9 10 U1 0 U2 0 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 0264-410X J9 VACCINE JI Vaccine PD JUN 14 PY 2001 VL 19 IS 27 BP 3620 EP 3631 DI 10.1016/S0264-410X(01)00101-3 PG 12 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 443AQ UT WOS:000169317500005 PM 11395195 ER PT J AU Stittelaar, KJ Kuiken, T de Swart, RL van Amerongen, G Vos, HW Niesters, HGM van Schalkwijk, P van der Kwast, T Wyatt, LS Moss, B Osterhaus, ADME AF Stittelaar, KJ Kuiken, T de Swart, RL van Amerongen, G Vos, HW Niesters, HGM van Schalkwijk, P van der Kwast, T Wyatt, LS Moss, B Osterhaus, ADME TI Safety of modified vaccinia virus Ankara (MVA) in immune-suppressed macaques SO VACCINE LA English DT Article DE modified vaccinia virus Ankara; smallpox; macaques ID T-CELL INDUCTION; RHESUS-MONKEYS; VIRAL REPLICATION; MEASLES; VACCINATION; INFECTION; STRAIN; HOST; IMMUNOGENICITY; IMMUNIZATION AB Modified vaccinia virus Ankara (MVA)-based recombinant viruses have been shown to be potent vaccine candidates for several infectious and neoplastic diseases. Since a major application of these live, replication-deficient vectors would be their use in immunocompromised or potentially immunocompromised individuals. a preclinical safety study was carried out. Macaques were inoculated with high doses of MVA (10(9)) via various routes. after immune-suppression by total-body irradiation, anti-thymocyte globulin treatment, or measles virus (MV) infection. No clinical, haematological or pathological abnormalities related to MVA inoculation were observed during a 13-day follow-up period. The presence of MVA genomes was demonstrated by nested PCR during the course of the experiment in ail macaques. but from none of these animals replication competent MVA could be reisolated. These data suggest that MVA can safely be used as a basis for recombinant human vaccines, and that it is also safe for use in immunocompromised individuals. (C) 2001 Elsevier Science Ltd. All rights reserved. C1 Erasmus Univ, Inst Virol, NL-3000 DR Rotterdam, Netherlands. Natl Inst Publ Hlth & Environm, NL-3720 BA Bilthoven, Netherlands. Erasmus Univ, Expt Anim Ctr, NL-3000 DR Rotterdam, Netherlands. NIAID, Viral Dis Lab, Bethesda, MD 20892 USA. RP Osterhaus, ADME (reprint author), Erasmus Univ, Inst Virol, POB 1738, NL-3000 DR Rotterdam, Netherlands. RI De Swart, Rik/E-6508-2011; OI De Swart, Rik/0000-0003-3599-8969; Niesters, Hubert/0000-0002-1758-0430 NR 35 TC 127 Z9 130 U1 0 U2 0 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 0264-410X J9 VACCINE JI Vaccine PD JUN 14 PY 2001 VL 19 IS 27 BP 3700 EP 3709 DI 10.1016/S0264-410X(01)00075-5 PG 10 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 443AQ UT WOS:000169317500014 PM 11395204 ER PT J AU Naficy, AB Trach, DD Ke, NT Chuc, NTK Sorkin, A Rao, MR Sy, TH Thiem, VD Canh, DG Mahoney, RT Holmgren, J Ivanoff, B Clemens, JD AF Naficy, AB Trach, DD Ke, NT Chuc, NTK Sorkin, A Rao, MR Sy, TH Thiem, VD Canh, DG Mahoney, RT Holmgren, J Ivanoff, B Clemens, JD TI Cost of immunization with a locally produced, oral cholera vaccine in Viet Nam SO VACCINE LA English DT Article DE cholera; immunization; cost ID FIELD TRIAL; DEVELOPING-COUNTRIES; FOLLOW-UP; BANGLADESH; EFFICACY; DISEASE AB Policy decisions regarding whether to incorporate new vaccines into routine public health practice in developing countries will depend in part on the costs of vaccine purchase and of vaccine delivery. In March, 1997, a large-scale effectiveness trial of a locally produced, orally administered bivalent vaccine against Vibrio cholerae 01 and 0139 began in Viet Nam. Empirical data obtained from the trial was used to determine the costs of the immunization campaign from the government perspective. The study population, including the children less than one year of age and pregnant women who were ineligible for immunization, was 353926. A total of 289041 persons received two doses of vaccine, and 13340 persons received one dose of vaccine. Two-dose vaccine coverage was 83.4%. The total cost of vaccine delivery during the immunization campaign was $66527. The cost of each dose of vaccine was $0.31. Therefore. the total cost of the immunization campaign was $0.44 per dose administered, and $0.91 per fully immunized person. Attempts to reduce the cost per dose of vaccine (e.g. the use of a monovalent vaccine against serogroup 01) are likely to have a large impact on the cost of future similar immunization campaigns. Published by Elevier Science Ltd. C1 NICHHD, Epidemiol Branch, Rockville, MD 20852 USA. Natl Inst Hyg & Epidemiol, Hanoi, Vietnam. Natl Inst Vaccines & Biol Subst, Nha Trang, Vietnam. Ctr Social Sci Hlth, Hanoi, Vietnam. Johns Hopkins Univ, Sch Hyg & Publ Hlth, Baltimore, MD USA. Univ Maryland Baltimore Cty, Catonsville, MD USA. Khanh Hoa Hlth Serv, Nha Trang, Vietnam. Int Vaccine Inst, Seoul, South Korea. Univ Gothenburg, Gothenburg, Sweden. WHO, CH-1211 Geneva, Switzerland. RP Naficy, AB (reprint author), NICHHD, Epidemiol Branch, 6100 Execut Blvd,Suite 7B03, Rockville, MD 20852 USA. RI Mahoney, Richard/B-4971-2009 NR 17 TC 15 Z9 15 U1 0 U2 1 PU ELSEVIER SCI LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, OXON, ENGLAND SN 0264-410X J9 VACCINE JI Vaccine PD JUN 14 PY 2001 VL 19 IS 27 BP 3720 EP 3725 DI 10.1016/S0264-410X(01)00089-5 PG 6 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 443AQ UT WOS:000169317500016 PM 11395206 ER PT J AU Leasure, CS Chandler, J Gilbert, DJ Householder, DB Stephens, R Copeland, NG Jenkins, NA Sharan, SK AF Leasure, CS Chandler, J Gilbert, DJ Householder, DB Stephens, R Copeland, NG Jenkins, NA Sharan, SK TI Sequence, chromosomal location and expression analysis of the murine homologue of human RAD51L2/RAD51C SO GENE LA English DT Article DE TestCode; genscan; 5 ' RACE; DNA repair; bacterial artificial chromosome (BAC) ID EMBRYONIC LETHALITY; MAMMALIAN-CELLS; BREAST-CANCER; RECA HOMOLOGS; RAD51; DNA; RECOMBINATION; REPAIR; MOUSE; SYNAPSIS AB The Rad51. protein has been shown to playa vital role in the DNA repair process. In humans, its interaction with proteins like BRCA1 and BRCA2 has provided an insight into the mechanism of how these molecules function as tumor suppressors. Several members of the Rad51 like family have been recently identified, including RAD51L2. This gene has been found to be amplified in breast tumors suggesting its role in tumor progression. Here, we describe the cloning of the murine homologue of the human RAD51L2/RAD51C gene. Sequence analysis has revealed that the murine Rad5112 protein is 86% identical and 93% similar to its human homologue. In spite of such high sequence conservation, the murine protein lacks the first nine amino acids present in the human protein. We have cloned and confirmed the sequence of the 5 ' end of the murine Rad5112 cDNA using 5 ' RACE technique as well as by sequencing the genomic region flanking the first exon of the murine Rad5112 gene. Northern analysis shows that Rad5112 is expressed in several adult tissues as well as in embryos at various developmental stages. The murine Rad5112 gene maps to chromosome 11 and is located in the syntenic region of human chromosome 17q22-23, where the human RAD51L2 is present. Published by Elsevier Science B.V. C1 NCI, Mouse Canc Genet Program, Ft Detrick, MD 21702 USA. NCI, SAIC Frederick, Adv Biomed Comp Ctr, Frederick, MD 21702 USA. RP Sharan, SK (reprint author), NCI, Mouse Canc Genet Program, Bldg 538,Room 133,POB B, Ft Detrick, MD 21702 USA. FU NCI NIH HHS [N01-CO-56000] NR 35 TC 11 Z9 14 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 JUN 13 PY 2001 VL 271 IS 1 BP 59 EP 67 DI 10.1016/S0378-1119(01)00498-X PG 9 WC Genetics & Heredity SC Genetics & Heredity GA 460VL UT WOS:000170329200007 PM 11410366 ER PT J AU Stoler, MH Schiffman, M AF Stoler, MH Schiffman, M TI Interobserver agreement about cervical cytologic and histologic diagnosis - Reply SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Letter C1 Univ Virginia, Charlottesville, VA 22903 USA. NCI, Bethesda, MD 20892 USA. RP Stoler, MH (reprint author), Univ Virginia, Charlottesville, VA 22903 USA. NR 2 TC 3 Z9 3 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 JUN 13 PY 2001 VL 285 IS 22 BP 2855 EP 2856 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 440BP UT WOS:000169156300019 ER PT J AU Srinivasan, A Kebede, N Saavedra, JE Nikolaitchik, AV Brady, DA Yourd, E Davies, KM Keefer, LK Toscano, JP AF Srinivasan, A Kebede, N Saavedra, JE Nikolaitchik, AV Brady, DA Yourd, E Davies, KM Keefer, LK Toscano, JP TI Chemistry of the diazeniumdiolates. 3. Photoreactivity SO JOURNAL OF THE AMERICAN CHEMICAL SOCIETY LA English DT Article ID NITRIC-OXIDE DONORS; ORTHO-NITROBENZYL ESTERS; LONG-TERM POTENTIATION; NITROSO OXIDES; CAGED COMPOUNDS; PHOTOCHEMISTRY; RELEASE; NO; OXYGEN; SPECTROSCOPY AB We have found O-2-substituted diazeniumdiolates, compounds of structure R2N-N(O)=NOR ' that are under development for various possible pharmaceutical uses, to be rather photosensitive. With R = ethyl and R ' = methyl, benzyl, or 2-nitrobenzyl, the observed product distributions suggest that two primary pathways are operative. A minor pathway involves the extrusion of nitrous oxide (N2O) with simultaneous generation of R2N(.) and R 'O-., which may then form amines, aldehydes, and alcohols. The major reaction pathway is an interesting photochemical cleavage of the N double bondN bond to form a nitrosamine (R2NN double bondO) and an oxygen-substituted nitrene (R ' ON). The intermediacy of the O-nitrene was inferred from the production of abundant oxime, via rearrangement of the O-nitrene to a C-nitroso compound (R ' ON --> O double bond NR '), and subsequent tautomerization to the more stable oxime. Involvement of the O-nitrene was confirmed by trapping with 2,3dimethyl-2-butene to form the aziridine and with oxygen to generate the nitrate ester. 2-Nitro substitution on the benzyl derivative had surprisingly little effect on the reaction course. For each compound examined, minor amounts of nitric oxide (NO), presumably produced by secondary photolysis of the nitrosamine, were observed. Time-resolved infrared experiments provided additional support for the above reaction pathways and confirmed that the nitrosamine is a primary photoproduct. We have also found that the relative contributions of the reaction pathways can be altered in certain derivatives. For example, when R ' = 2,4-dinitrophenyl, the contribution of the nitrosamine/O-nitrene-forming pathway was diminished. Pharmacological implications of these results are discussed. C1 NCI, Comparat Carcinogenesis Lab, Chem Sect, Frederick, MD 21702 USA. Johns Hopkins Univ, Dept Chem, Baltimore, MD 21218 USA. NCI, SAIC Frederick, Intramural Res Support Program, Frederick, MD 21702 USA. George Mason Univ, Dept Chem, Fairfax, VA 22030 USA. RP Toscano, JP (reprint author), NCI, Comparat Carcinogenesis Lab, Chem Sect, Frederick, MD 21702 USA. RI Keefer, Larry/N-3247-2014 OI Keefer, Larry/0000-0001-7489-9555 FU NCI NIH HHS [N01-CO-56000]; NIGMS NIH HHS [R01 GM58109] NR 79 TC 50 Z9 50 U1 1 U2 12 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 JUN 13 PY 2001 VL 123 IS 23 BP 5465 EP 5472 DI 10.1021/ja002898y PG 8 WC Chemistry, Multidisciplinary SC Chemistry GA 440LG UT WOS:000169176300013 PM 11389628 ER PT J AU Davies, KM Wink, DA Saavedra, JE Keefer, LK AF Davies, KM Wink, DA Saavedra, JE Keefer, LK TI Chemistry of the diazeniumdiolates. 2. Kinetics and mechanism of dissociation to nitric oxide in aqueous solution SO JOURNAL OF THE AMERICAN CHEMICAL SOCIETY LA English DT Article ID BIOLOGICAL-SYSTEMS; NO; PIPERAZINE; COMPLEXES; RELEASE AB Diazeniumdiolate ions of structure R2N[N(O)NO](-) (1) are of pharmacological interest because they spontaneously generate the natural bioregulatory species, nitric oxide (NO), when dissolved in aqueous media. Here we report the kinetic details for four representative reactivity patterns: (a) straightforward dissociation of the otherwise unfunctionalized diethylamine derivative 2 (anion 1, where R = Et) to diethylamine and NO; (b) results for the zwitterionic piperazin-1-yl analogue 4, for which the protonation state of the neighboring basic amine site is an important determinant of dissociation rate; (c) data for 5, a diazeniumdiolate derived from the polyamine spermine, whose complex rate equation can include terms for a variety of medium effects; and (d) the outcome for triamine 6 (R = CH2CH2NH3+), the most stable structure 1 ion identified to date. All of these dissociations are acid-catalyzed, with equilibrium protonation of the substrate preceding release of NO. Specific rate constants and pK(a) values for 2-6 have been determined from pH/rate profiles. Additionally, a hypsochromic shift (from similar to 250 to similar to 230 nm) was observed on acidifying these ions, allowing determination of a separate pK(a) for each substrate. For 6, the pK(a) value obtained kinetically was 2-3 pK(a) units higher than the value obtained from the spectral shift. Comparison of the ultraviolet spectra for 6 at various pH values with those for O- and N-alkylated diazeniumdiolates suggests that protonation at the R2N nitrogen initiates dissociation to NO at physiological pH, with a second protonation (at oxygen) accounting for both the spectral change and the enhanced dissociation rate at pH <4. Our results help to explain the previously noted variability in dissociation rate of 5, whose half-life we found to increase by an order of magnitude when its concentration was raised from near-zero to 1 mM, and provide mechanistic insight into the factors that govern dissociation rates among diazeniumdiolates of importance as pharmacologic progenitors of NO. C1 George Mason Univ, Dept Chem, Fairfax, VA 22030 USA. NCI, Comparat Carcinogenesis Lab, Chem Sect, Frederick, MD 21702 USA. NCI, SAIC Frederick, Intramural Res Support Program, Frederick, MD 21702 USA. RP Davies, KM (reprint author), George Mason Univ, Dept Chem, Fairfax, VA 22030 USA. RI Keefer, Larry/N-3247-2014 OI Keefer, Larry/0000-0001-7489-9555 FU NCI NIH HHS [N01-CO-56000] NR 34 TC 145 Z9 146 U1 0 U2 17 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 JUN 13 PY 2001 VL 123 IS 23 BP 5473 EP 5481 DI 10.1021/ja002899q PG 9 WC Chemistry, Multidisciplinary SC Chemistry GA 440LG UT WOS:000169176300014 PM 11389629 ER PT J AU Friso, S Jacques, PF Wilson, PWF Rosenberg, IH Selhub, J AF Friso, S Jacques, PF Wilson, PWF Rosenberg, IH Selhub, J TI Low circulating vitamin B-6 is associated with elevation of the inflammation marker C-reactive protein independently of plasma homocysteine levels SO CIRCULATION LA English DT Article DE atherosclerosis; risk factors; homocysteine; vitamins; inflammation ID CORONARY-ARTERY DISEASE; RHEUMATOID-ARTHRITIS; RISK-FACTORS; ATHEROSCLEROSIS; DEFICIENCY; FOLATE; STROKE; ASSAY AB Background - Lower vitamin B, concentrations are reported to confer an increased and independent risk for cardiovascular disease (CVD). The mechanism underlying this relationship, however, remains to be defined. Other diseases, such as rheumatoid arthritis, are associated with reduced vitamin B, levels. Despite a clear distinction in pathophysiology, inflammatory reaction may be the major link between these diseases. We hypothesized a relationship between pyridoxal 5 ' -phosphate (PLP), the active form of vitamin B,, and the marker of inflammation C-reactive protein (CRP). We also evaluated whether total plasma homocysteine (tHcy), a well-defined risk factor for CVD and a major determinant of plasma PLP levels, had a possible role as a mediator of this hypothesized relationship. Methods and Results - Data from 891 participants from the population-based Framingham Heart Study cohort were analyzed. Subjects were divided into 2 groups according to normal or elevated CRP values: group 1, CRP <6 mg/L; group 2, CRP greater than or equal to6 mg/L. Plasma PLP levels were substantially lower in group 2 than in group 1 (mean values in group 2, 36.5 nmol/L versus 55.8 nmol/L in group 1, P <0.001). In a multiple logistic regression model adjusted for tHcy, the association of PLP with CRP remained highly significant (P=0.003). Conclusions - Low plasma PLP is associated with higher CRP levels independently of tHcy. This observation may reflect a vitamin B-6 utilization in the presence of an underlying inflammatory process and represent a possible mechanism to explain the decreased vitamin B-6 levels in CVD. C1 NHLBI, Framingham Heart Study, Framingham, MA USA. Boston Univ, Sch Med, Boston, MA 02118 USA. RP Selhub, J (reprint author), Tufts Univ, Jean Mayer USDA, Human Nutr Res Ctr, 711 Washington St, Boston, MA 02111 USA. NR 28 TC 148 Z9 149 U1 0 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 JUN 12 PY 2001 VL 103 IS 23 BP 2788 EP 2791 PG 4 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 443RF UT WOS:000169353600016 PM 11401933 ER PT J AU Lahat, H Eldar, M Levy-Nissenbaum, E Bahan, T Friedman, E Khoury, A Lorber, A Kastner, DL Goldman, B Pras, E AF Lahat, H Eldar, M Levy-Nissenbaum, E Bahan, T Friedman, E Khoury, A Lorber, A Kastner, DL Goldman, B Pras, E TI Autosomal recessive catecholamine- or exercise-induced polymorphic ventricular tachycardia - Clinical features and assignment of the disease gene to chromosome 1p13-21 SO CIRCULATION LA English DT Article DE tachycardia; genetics; mapping; death, sudden; syncope ID LOCALIZATION; RECEPTOR; HEART AB Background-Catecholaminergic polymorphic ventricular tachycardia (PVT) is characterized by episodes of syncope, seizures, or sudden death in response to physiological or emotional stress. In 2 families with autosomal dominant inheritance, the disease gene was mapped to chromosome 1q42-43. The objectives of this study were to characterize the clinical features of the disease in a Bedouin tribe from Israel and to map the disease gene. Methods and Results - In this Bedouin tribe, 9 children (age, 7 +/-4 years) from 7 related families have died suddenly during the past decade, and 12 other children suffered from recurrent syncope and seizures starting at the age of 6 +/-3 years. Parents of affected individuals were asymptomatic and were all related (first-, second-, or third-degree cousins), Segregation analysis suggested autosomal recessive inheritance. All 12 symptomatic patients and 1 asymptomatic sibling (mean age, 13 +/-7 years) were found to have a relative resting bradycardia (64 +/- 13 bpm, versus 930 +/- 12 bpm in the unaffected siblings), as well as PVT induced by treadmill or isoproterenol infusion and appearing at a mean sinus rate of 110 +/-0 bpm. Patients responded favorably to treatment with P-blockers, A genome-wide search using polymorphic DNA markers mapped the disease locus to a 16-megabase interval on chromosome 1p13-21. A maximal lod score of 8.24 was obtained with D1S189 at theta =0.00, Sequencing of KCND3, a gene that encodes an I-tO potassium channel transporter, did not reveal any significant sequence alterations. Conclusions - This unique form of autosomal recessive PVT affects young children and may be lethal if left untreated. Linkage analysis maps this disorder to chromosome 1p13-21. C1 Tel Aviv Univ, Sackler Sch Med, Chaim Sheba Med Ctr, Danek Gartner Inst Human Genet, IL-52621 Tel Hashomer, Israel. Tel Aviv Univ, Sackler Sch Med, Chaim Sheba Med Ctr, Neufeld Cardiac Res Inst, IL-52621 Tel Hashomer, Israel. Tel Aviv Univ, Sackler Sch Med, Chaim Sheba Med Ctr, Susanne Levy Gartner Oncogenet Unit, IL-52621 Tel Hashomer, Israel. Rambam Med Ctr, Inst Pediat Cardiol, Haifa, Israel. NIAMSD, Genet Sect, NIH, Bethesda, MD 20892 USA. RP Pras, E (reprint author), Tel Aviv Univ, Sackler Sch Med, Chaim Sheba Med Ctr, Danek Gartner Inst Human Genet, IL-52621 Tel Hashomer, Israel. NR 16 TC 142 Z9 151 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 JUN 12 PY 2001 VL 103 IS 23 BP 2822 EP 2827 PG 6 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 443RF UT WOS:000169353600022 PM 11401939 ER PT J AU Padayatty, SJ Levine, M AF Padayatty, SJ Levine, M TI Vitamin C and coronary microcirculation SO CIRCULATION LA English DT Letter ID TRANSPORTERS C1 NIDDK, Mol & Clin Nutr Sect, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. RP Levine, M (reprint author), NIDDK, Mol & Clin Nutr Sect, Digest Dis Branch, NIH, Bethesda, MD 20892 USA. RI Padayatty, Sebastian/A-8581-2012 OI Padayatty, Sebastian/0000-0001-8758-3170 NR 4 TC 9 Z9 9 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 JUN 12 PY 2001 VL 103 IS 23 BP E117 EP E117 PG 1 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 443RF UT WOS:000169353600002 PM 11401949 ER PT J AU Higgins, JJ Loveless, JM Goswami, S Nee, LE Cozzo, C De Biase, A Rosen, DR AF Higgins, JJ Loveless, JM Goswami, S Nee, LE Cozzo, C De Biase, A Rosen, DR TI An atypical intronic deletion widens the spectrum of mutations in hereditary spastic paraplegia SO NEUROLOGY LA English DT Article ID MULTILOCUS LINKAGE ANALYSIS; LOCUS; MAPS AB Objective: To identify the genetic mutation responsible for autosomal dominant spastic paraplegia (HSP) in a large family with a "pure" form of the disorder. Background: The disease locus in most families with HSP is genetically linked to the SPG4 locus on chromosome 2p21-p22. Some of these families have mutations in the splice-site or coding regions of the spastin gene (SPAST). Methods: Linkage and mutational analyses were used to identify the location and the nature of the genetic defect causing the disorder in a large family. After the disease phenotype was linked to the SPG4 locus, all 17 coding regions and flanking intronic sequences of SPAST were analyzed by single-strand conformation polymorphism analysis (SSCP) and compared between affected and normal individuals. Direct sequencing and subcloning methods were used to investigate incongruous mobility shifts. Results: The genomic sequence of SPAST showed a heterozygous four-base pair deletion (deITAAT) near the 3 ' splice-site of exon three in all 11 affected individuals but not in 21 normal family members or in 50 unrelated controls (100 chromosomes). Conclusions: This study identifies an atypical intronic microdeletion in SPAST that causes HSP and widens the spectrum of genetic abnormalities that cause the disorder. C1 New York State Dept Hlth, Wadsworth Ctr, Neurogenet Lab, Albany, NY USA. New York State Dept Hlth, Wadsworth Ctr, Lab Neuromuscular Genet, Albany, NY USA. NIH, Natl Inst Neurol Disorders & Stroke, Family Studies Unit, Bethesda, MD USA. RP Higgins, JJ (reprint author), Westchester Med Ctr, Mid Hudson Family Hlth Inst, Ctr Human Genet Studies, 279 Main St,Suite 202, New Paltz, NY 12561 USA. FU NINDS NIH HHS [NS39353, NS35932] NR 18 TC 14 Z9 14 U1 0 U2 1 PU LIPPINCOTT WILLIAMS & WILKINS PI PHILADELPHIA PA 530 WALNUT ST, PHILADELPHIA, PA 19106-3621 USA SN 0028-3878 J9 NEUROLOGY JI Neurology PD JUN 12 PY 2001 VL 56 IS 11 BP 1482 EP 1485 PG 4 WC Clinical Neurology SC Neurosciences & Neurology GA 440RB UT WOS:000169187100010 PM 11402104 ER PT J AU Worzala, K Hiller, R Sperduto, RD Mutalik, K Murabito, JM Moskowitz, M D'Agostino, RB Wilson, PWF AF Worzala, K Hiller, R Sperduto, RD Mutalik, K Murabito, JM Moskowitz, M D'Agostino, RB Wilson, PWF TI Postmenopausal estrogen use, type of menopause, and lens opacities - The Framingham studies SO ARCHIVES OF INTERNAL MEDICINE LA English DT Article ID BEAVER DAM EYE; HORMONE REPLACEMENT THERAPY; AGE-RELATED CATARACTS; BLUE MOUNTAINS EYE; BODY-MASS INDEX; CIGARETTE-SMOKING; RISK-FACTORS; SUBCAPSULAR CATARACTS; PREVALENCE; WOMEN AB Background: Previous studies of estrogen replacement therapy and lens opacities have not reported consistent findings. Objective: To investigate whether postmenopausal estrogen use is associated with the occurrence of age-related lens opacities (nuclear, cortical, and posterior subcapsular). Methods: Surviving members of the original cohort of the Framingham Heart Study who also participated in the Framingham Eye Study (1986-1989) were examined for the absence or presence of lens opacities. Data from the Framingham Heart Study, including information on menopausal status (collected biennially from approximately 1948) and use of estrogen replacement therapy (collected biennially from approximately 1960) were used to examine associations between lens opacities and duration of postmenopausal estrogen use, type of menopause, and age at menopause. Five hundred twenty-nine women, aged 66 to 93 years, were included. Multivariable-adjusted odds ratios of specific types of lens opacities were calculated for (1) duration of estrogen use (never and 1-2, 3-9, and greater than or equal to 10 years), (2) surgical vs natural menopause, and (3) age at menopause. Results: Longer duration of postmenopausal estrogen therapy was inversely associated with the presence of nuclear lens opacities in an adjusted model. Women who had taken estrogen for 10 years or longer had a 60% reduction in risk compared with nonusers (odds ratio, 0.4; 95% confidence interval, 0.2-1.01). Longer duration of estrogen use was associated with fewer posterior subcapsular opacities at a borderline level of significance. No association was noted fur cortical opacities. The risk of posterior subcapsular opacities was significantly increased for women who had undergone surgical menopause compared with women with natural menopause (odds ratio, 2.2; 95% confidence interval, 1.1-4.3). No association was noted for lens opacities and age at menopause. Conclusion: Data from our study and other studies suggest that a reduction in the risk of lens opacities may be an additional benefit of postmenopausal estrogen use. C1 Boston Univ, Dept Math & Stat, Boston, MA 02215 USA. Boston Univ, Endocrinol Sect, Boston, MA 02215 USA. Boston Univ, Gen Internal Med Sect, Boston, MA 02215 USA. NHLBI, Framingham, MA USA. NEI, Div Epidemiol & Clin Res, Bethesda, MD 20892 USA. Univ Minnesota, Sch Med, Gen Internal Med Sect, Vet Affairs Med Ctr, Minneapolis, MN 55455 USA. RP Hiller, R (reprint author), NEI, Div Epidemiol & Clin Res, Bldg 31,Room 6A52-31 Ctr Dr MSC 2510, Bethesda, MD 20892 USA. FU NEI NIH HHS [N01-EY-2-2112, N01-EY-6-2105] NR 41 TC 49 Z9 56 U1 0 U2 3 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0003-9926 J9 ARCH INTERN MED JI Arch. Intern. Med. PD JUN 11 PY 2001 VL 161 IS 11 BP 1448 EP 1454 DI 10.1001/archinte.161.11.1448 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 439UR UT WOS:000169136400012 PM 11386895 ER PT J AU Nechushtan, A Smith, CL Lamensdorf, I Yoon, SH Youle, RJ AF Nechushtan, A Smith, CL Lamensdorf, I Yoon, SH Youle, RJ TI Bax and Bak coalesce into novel mitochondria-associated clusters during apoptosis SO JOURNAL OF CELL BIOLOGY LA English DT Article DE apoptosis; Bcl-2; green fluorescent protein; mitochondria; clusters ID CYTOCHROME-C RELEASE; BCL-X-L; INDUCED CONFORMATIONAL CHANGE; PERMEABILITY TRANSITION PORE; CHANNEL-FORMING ACTIVITY; PROGRAMMED CELL-DEATH; SUBCELLULAR-LOCALIZATION; ION-CHANNEL; IN-VIVO; PROTEIN AB Bax is a member of the Bcl-2 family of proteins known to regulate mitochondria-dependent programmed cell death. Early in apoptosis, Bax translocates from the cytosol to the mitochondrial membrane. We have identified by confocal and electron microscopy a novel step in the Bax proapoptotic mechanism immediately subsequent to mitochondrial translocation. Bax leaves the mitochondrial membranes and coalesces into large clusters containing thousands of Bax molecules that remain adjacent to mitochondria, Bak, a close homologue of Bax, colocalizes in these apoptotic clusters in contrast to other family members, Bid and Bad, which circumscribe the outer mitochondrial membrane throughout cell death progression. We found the formation of Bax and Bak apoptotic clusters to be caspase independent and inhibited completely and specifically by Bcl-X-L, correlating cluster formation with cytotoxic activity. Our results reveal the importance of a novel structure formed by certain Bcl-2 family members during the process of cell death. C1 NINCDS, Surg Neurol Branch, Biochem Sect, NIH, Bethesda, MD 20892 USA. NINCDS, Light Imaging Facil, NIH, Bethesda, MD 20892 USA. NINCDS, Clin Neurosci Branch, NIH, Bethesda, MD 20892 USA. RP Youle, RJ (reprint author), NINCDS, Surg Neurol Branch, Biochem Sect, NIH, 10 Ctr Dr,Rm 5D37, Bethesda, MD 20892 USA. NR 50 TC 337 Z9 358 U1 1 U2 8 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 1114 FIRST AVE, 4TH FL, NEW YORK, NY 10021 USA SN 0021-9525 J9 J CELL BIOL JI J. Cell Biol. PD JUN 11 PY 2001 VL 153 IS 6 BP 1265 EP 1276 DI 10.1083/jcb.153.6.1265 PG 12 WC Cell Biology SC Cell Biology GA 443AX UT WOS:000169318100013 PM 11402069 ER PT J AU Speransky, VV Taylor, KL Edskes, HK Wickner, RB Steven, AC AF Speransky, VV Taylor, KL Edskes, HK Wickner, RB Steven, AC TI Prion filament networks in [URE3] cells of Saccharomyces cerevisiae SO JOURNAL OF CELL BIOLOGY LA English DT Article DE amyloid; yeast prion; immunoelectron microscopy; protease resistance; Ure2p ID YEAST PRION; PROTEIN HSP104; SCRAPIE PRION; IN-VITRO; DOMAIN; PROPAGATION; DETERMINANT; PSI(+); SUP35; GENE AB The [URE3] prion (infectious protein) of yeast is a self-propagating, altered form of Ure2p that cannot carry out its normal function in nitrogen regulation. Previous data have shown that Ure2p can form protease-resistant amyloid filaments in vitro, and that it is aggregated in cells carrying the [URE3] prion. Here we show by electron microscopy that [URE3] cells overexpressing Ure2p contain distinctive, filamentous networks in their cytoplasm, and demonstrate by immunolabeling that these networks contain Ure2p. In contrast, overexpressing wild-type cells show a variety of Ure2p distributions: usually, the protein is dispersed sparsely throughout the cytoplasm, although occasionally it is found in multiple small, focal aggregates. However, these distributions do not resemble the single, large networks seen in [URE3] cells, nor do the control cells exhibit cytoplasmic filaments. In [URE3] cell extracts, Ure2p is present in aggregates that are only partially solubilized by boiling in SDS and urea. In these aggregates, the NH2-terminal prion domain is inaccessible to antibodies, whereas the COOH-terminal nitrogen regulation domain is accessible. This finding is consistent with the proposal that the prion domains stack to form the filament backbone, which is surrounded by the COOH-terminal domains. These observations support and further specify the concept of the [URE3] prion as a self-propagating amyloid. C1 NIAMSD, Lab Struct Biol, NIH, Bethesda, MD 20892 USA. NIDDKD, Lab Biochem & Genet, NIH, Bethesda, MD 20892 USA. RP NIAMSD, Lab Struct Biol, NIH, Bldg 6,Room B2-34,6 Ctr Dr,MSC 2717, Bethesda, MD 20892 USA. EM steven@calvin.niams.nih.gov NR 33 TC 68 Z9 69 U1 0 U2 2 PU ROCKEFELLER UNIV PRESS PI NEW YORK PA 950 THIRD AVE, 2ND FLR, NEW YORK, NY 10022 USA SN 0021-9525 EI 1540-8140 J9 J CELL BIOL JI J. Cell Biol. PD JUN 11 PY 2001 VL 153 IS 6 BP 1327 EP 1335 DI 10.1083/jcb.153.6.1327 PG 9 WC Cell Biology SC Cell Biology GA 443AX UT WOS:000169318100018 PM 11402074 ER PT J AU Gurbuxani, S Arya, LS Raina, V Sazawal, S Khattar, A Magrath, I Marie, JP Bhargava, M AF Gurbuxani, S Arya, LS Raina, V Sazawal, S Khattar, A Magrath, I Marie, JP Bhargava, M TI Significance of MDR1, MRP1, GST pi and GST mu mRNA expression in acute lymphoblastic leukemia in Indian patients SO CANCER LETTERS LA English DT Article DE Indian acute lymphoblastic leukemia; MRP1; mechanisms of multidrug resistance 1; glutathione S-transferase pi; glutathione S-transferase mu; reverse transcription-polymerase chain reaction ID RESISTANCE-ASSOCIATED PROTEIN; ACUTE MYELOID-LEUKEMIA; CELLULAR-DRUG RESISTANCE; S-TRANSFERASE-PI; MULTIDRUG-RESISTANCE; GLYCOPROTEIN P-170; PROGNOSTIC FACTORS; GENE-EXPRESSION; GLUTATHIONE; CELLS AB Using, semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) in 167 patients of acute lymphoblastic leukemia (ALL) from India at different stages of the disease (presentation 125, remission 33, first relapse nine), MRP1 and GST pi expression were significantly higher at relapse than presentation (P = 0.03 and P = 0.01, respectively) and remission (P = 0.007 and P = 0.003, respectively). MRPI, GST pi and GST mu, were expressed simultaneously in several samples with significant association of expression levels (P = 0.0001). Association with clinicopathological features included higher MDR1 expression with age >15 years (P = 0.04) and higher MRP1, GST pi, GST mu expression with WBC counts >100 x 10(9)/l. In 71 patients (age <25 years), inability to achieve CR was associated with a significantly higher MDR1 mRNA expression (P = 0.03) indicating a prognostic significance. However, relapse or shorter Event Free Survival was independent of mRNA expression levels of the four genes. In view of the increased mRNA expression of MRP1/GST at the time of relapse and an association with risk factors such as a high WBC count, further studies directed towards investigating the functional aspects of GSH/GST/MRP1 mediated drug transport are warranted. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved. C1 All India Inst Med Sci, Dept Haematol, New Delhi 110029, India. NCI, Lymphoma Biol Sect, Paediat Branch, NIH, Bethesda, MD 20892 USA. Hosp Hotel Dieu Paris, Dept Haematol, Paris, France. RP Bhargava, M (reprint author), All India Inst Med Sci, Dept Haematol, New Delhi 110029, India. OI Bhargava, Manorama/0000-0003-2135-7298 NR 41 TC 5 Z9 8 U1 0 U2 1 PU ELSEVIER SCI IRELAND LTD PI CLARE PA CUSTOMER RELATIONS MANAGER, BAY 15, SHANNON INDUSTRIAL ESTATE CO, CLARE, IRELAND SN 0304-3835 J9 CANCER LETT JI Cancer Lett. PD JUN 10 PY 2001 VL 167 IS 1 BP 73 EP 83 DI 10.1016/S0304-3835(00)00684-4 PG 11 WC Oncology SC Oncology GA 435XN UT WOS:000168907600010 PM 11323101 ER PT J AU Mahato, D Goulding, EH Korach, KS Eddy, EM AF Mahato, D Goulding, EH Korach, KS Eddy, EM TI Estrogen receptor-alpha is required by the supporting somatic cells for spermatogenesis SO MOLECULAR AND CELLULAR ENDOCRINOLOGY LA English DT Article; Proceedings Paper CT International Workshop on Estrogens and Male Reproduction CY SEP 23-24, 1999 CL CROTONE, ITALY DE estrogen receptor; spermatogenesis; germ-cell transplantation; epididymis ID MESSENGER-RIBONUCLEIC-ACID; REPRODUCTIVE-TRACT; WILD-TYPE; SPERMATOGONIAL TRANSPLANTATION; EFFERENT DUCTULES; BETA EXPRESSION; KNOCKOUT MICE; ER-ALPHA; MOUSE; TESTIS AB The gene for estrogen receptor-alpha (ER alpha) was disrupted ill embryonic stem cells by homologous recombination and these cells were used to generate mice with a targeted mutation in the ER alpha gene (alpha ERKO mice). It was found that males homozygous for the mutation are infertile, indicating that estrogen signaling through this nuclear hormone receptor is required for male reproductive function. Although spermatogenesis arrears normal in juvenile and young adult alpha ERKO mice, the sperm produced are unable to fertilize eggs in vitro. To determine whether ER alpha is required by somatic or germ cells in the male reproductive tract, we transplanted germ cells from homozygous mutant (ER alpha (-/-)) males to the testes of wild-type (ER alpha (+/+)) males depleted of germ cells by busulfan treatment. The recipients ('surrogate fathers') sired offspring heterozygous for the mutation (ER alpha ('/-) and carrying the coat-color marker of the infertile donor males. This indicated that ER alpha (-/-) germ cells an able to produce sperm competent to fertilize when they are supported by ER alpha (+/+). somatic cells. When ER alpha (+/-) offspring produced by germ cell transplantation were mated to produce ER alpha (-/-) males, these mice were found to have the same phenotype as originally reported for alpha ERKO males. These studies showed that male germ cells do not require ER alpha for regulation of their own genes for development and Function, and strongly imply that somatic cells of the male reproductive tract require ER alpha to support the production of sperm that are capable of fertilization. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved. C1 NIEHS, Reprod & Dev Toxicol Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Eddy, EM (reprint author), NIEHS, Reprod & Dev Toxicol Lab, NIH, MD C4-01,POB 12233, Res Triangle Pk, NC 27709 USA. OI Korach, Kenneth/0000-0002-7765-418X NR 38 TC 40 Z9 42 U1 0 U2 1 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 JUN 10 PY 2001 VL 178 IS 1-2 BP 57 EP 63 DI 10.1016/S0303-7207(01)00410-5 PG 7 WC Cell Biology; Endocrinology & Metabolism SC Cell Biology; Endocrinology & Metabolism GA 445XN UT WOS:000169484000009 PM 11403895 ER PT J AU Emanuel, EJ AF Emanuel, EJ TI Euthanasia: where the Netherlands leads will the world follow? No. legalisation is a diversion from improving care for the dying SO BRITISH MEDICAL JOURNAL LA English DT Editorial Material ID PHYSICIAN-ASSISTED SUICIDE C1 NIH, Warren G Magnuson Clin Ctr, Dept Clin Bioeth, Bethesda, MD 20892 USA. RP Emanuel, EJ (reprint author), NIH, Warren G Magnuson Clin Ctr, Dept Clin Bioeth, Bethesda, MD 20892 USA. NR 5 TC 14 Z9 14 U1 0 U2 2 PU BRITISH MED JOURNAL PUBL GROUP PI LONDON PA BRITISH MED ASSOC HOUSE, TAVISTOCK SQUARE, LONDON WC1H 9JR, ENGLAND SN 0959-535X J9 BRIT MED J JI Br. Med. J. PD JUN 9 PY 2001 VL 322 IS 7299 BP 1376 EP 1377 DI 10.1136/bmj.322.7299.1376 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 442KN UT WOS:000169284600002 PM 11397727 ER PT J AU Egan, CA Lazarova, Z Darling, TN Yee, C Cote, T Yancey, KB AF Egan, CA Lazarova, Z Darling, TN Yee, C Cote, T Yancey, KB TI Anti-epiligrin cicatricial pemphigoid and relative risk for cancer SO LANCET LA English DT Article AB It is not known whether patients with anti-epiligrin cicatricial pemphigoid (AECP) have an increased risk of malignancy. We calculated the expected numbers of cancers in a cohort of 35 such patients based on respective incidence rates for all cancers In the National Cancer Institute's Surveillance, Epidemiology, and End Results (NCI SEER) Registry. Ten patients in this cohort had solitary solid cancers; eight patients developed cancer after onset of AECP (seven within 14 months). The relative risk (RR) for cancer in this cohort was 6.8 (95% confidence intervals [CI]: 3.3-12.5). AECP seems to be associated with an increased relative risk for cancer. C1 NCI, Dermatol Branch, Div Clin Sci, NIH, Bethesda, MD 20892 USA. NCI, Canc Stat Branch, NIH, Rockville, MD USA. RP Egan, CA (reprint author), NCI, Dermatol Branch, Div Clin Sci, NIH, Bldg 10 Room 12N238, Bethesda, MD 20892 USA. OI Darling, Thomas/0000-0002-5161-1974 NR 5 TC 95 Z9 96 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 JUN 9 PY 2001 VL 357 IS 9271 BP 1850 EP 1851 DI 10.1016/S0140-6736(00)04971-0 PG 2 WC Medicine, General & Internal SC General & Internal Medicine GA 440DN UT WOS:000169160800016 PM 11410196 ER PT J AU Backman, C Hoffer, BJ Misawa, H Morales, M AF Backman, C Hoffer, BJ Misawa, H Morales, M TI Cellular mRNA expression of the transcription factor NGFI-B suggests a gene regulatory role in striatal opiate-peptide neurons SO BRAIN RESEARCH LA English DT Article DE NGFI-B; Nur77; enkephalin; dynorphin; striatum; distribution ID FACTOR-INDUCIBLE-B; ORPHAN NUCLEAR RECEPTORS; IMMEDIATE-EARLY GENES; MESSENGER-RNAS; RAT STRIATUM; BASAL GANGLIA; SUBSTANCE-P; DOPAMINE; ENKEPHALIN; PATTERNS AB Previous studies have shown that NGFI-B mRNA is highly expressed in the adult striatum. In the present study we analyzed the anatomical distribution of NGFI-B mRNA within this brain region as well as the degree of co-existence of NCFI-B with different striatal markers in the adult brain. NGFI-B mRNA levels were found to be significantly higher within the dorsomedial portion of the striatum as compared to the ventrolateral striatum. This distribution pattern was maintained throughout the rostro-caudal axis of the striatum. Double in situ hybridization studies showed that striatal NGFI-B mRNA colocalized with a subset of preproenkephalin and prodynorphin positive spiny neurons within the dorsomedial striatum; 22-28% of all opiate-peptide positive cells co-expressed NC;FI-D mRNA. NGFI-B did not colocalize with striatal aspiny interneurons expressing choline acetyl transferase mRNA or those containing the calcium-binding protein parvalbumin. The pattern of NGFI-B mRNA expression within different striatal spiny projecting neurons suggests that this transcription factor may have a direct effect on the function of different striatal efferent pathways. C1 NIDA, NIH, Baltimore, MD 21224 USA. Tokyo Metropolitan Inst Neurosci, Dept Neurol, Fuchu, Tokyo 183, Japan. RP Backman, C (reprint author), NIDA, NIH, 5500 Nathan Shock Dr, Baltimore, MD 21224 USA. RI backman, cristina/C-1276-2013 NR 28 TC 7 Z9 8 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0006-8993 J9 BRAIN RES JI Brain Res. PD JUN 8 PY 2001 VL 903 IS 1-2 BP 26 EP 32 DI 10.1016/S0006-8993(01)02332-0 PG 7 WC Neurosciences SC Neurosciences & Neurology GA 444NA UT WOS:000169404500003 PM 11382384 ER PT J AU Ghribi, O DeWitt, DA Forbes, MS Herman, MM Savory, J AF Ghribi, O DeWitt, DA Forbes, MS Herman, MM Savory, J TI Co-involvement of mitochondria and endoplasmic reticulum in regulation of apoptosis: changes in cytochrome c, Bcl-2 and Bax in the hippocampus of aluminum-treated rabbits SO BRAIN RESEARCH LA English DT Article DE Bcl-2; Bax; mitochondria; endoplasmic reticulum; Alzheimer's disease; aluminum ID PERMEABILITY TRANSITION PORE; NEUROFIBRILLARY DEGENERATION; FAMILY PROTEINS; CELL-DEATH; RELEASE; MECHANISM; BRAIN; IMMUNOREACTIVITY; SURVIVAL; LESIONS AB Neurodegenerative diseases, including Alzheimer's disease, are characterized by a progressive and selective loss of neurons. Apoptosis under mitochondrial control has been implicated in this neuronal death process, involving the release of cytochrome c into the cytoplasm and initiation of the apoptosis cascade. However. a growing body of evidence suggests an active role for the endoplasmic reticulum in regulating apoptosis, either independent of mitochondrial, or in concert with mitochondrial-initiated pathways. Members of the Bcl-2 family of proteins have been shown to either inhibit apoptosis, as is the case with Bcl-2, or to promote it, in the case of Bax. Investigations in our laboratory have focused on neuronal injury resulting from the intracisternal administration of aluminum maltolate to New Zealand white rabbits, an animal system relevant to a study of human disease in that it reflects many of the histological and biochemical changes associated with Alzheimer'.s disease. Here we report that treatment of young adult rabbits with aluminum maltolate induces both cytochrome c translocation into brain cytosol, and caspase-3 activation. Furthermore. as assessed by Western blot analysis, these effects are accompanied by a decrease in Bcl-2 and an increase in Bax reactivity in the endoplasmic reticulum. (C) 2001 Elsevier Science B.V. All rights reserved. C1 Univ Virginia, Hlth Sci Ctr, Dept Pathol, Charlottesville, VA 22908 USA. Univ Virginia, Hlth Sci Ctr, Dept Biochem & Mol Genet & Chem, Charlottesville, VA 22908 USA. Liberty Univ, Dept Biol & Chem, Lynchburg, VA USA. NIMH, IRP, NIH, Bethesda, MD 20892 USA. RP Savory, J (reprint author), Univ Virginia, Hlth Sci Ctr, Dept Pathol, Box 168, Charlottesville, VA 22908 USA. NR 39 TC 57 Z9 65 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0006-8993 J9 BRAIN RES JI Brain Res. PD JUN 8 PY 2001 VL 903 IS 1-2 BP 66 EP 73 DI 10.1016/S0006-8993(01)02406-4 PG 8 WC Neurosciences SC Neurosciences & Neurology GA 444NA UT WOS:000169404500008 PM 11382389 ER PT J AU Cabib, E Roh, DH Schmidt, M Crotti, LB Varma, A AF Cabib, E Roh, DH Schmidt, M Crotti, LB Varma, A TI The yeast cell wall and septum as paradigms of cell growth and morphogenesis SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Review ID GTP-BINDING PROTEIN; MAP KINASE PATHWAY; SACCHAROMYCES-CEREVISIAE; CHITIN SYNTHESIS; IDENTIFICATION; INTEGRITY; SYNTHASE; POLARIZATION; CHS3P; BIOSYNTHESIS C1 NIDDK, Lab Biochem & Genet, NIH, Bethesda, MD 20892 USA. RP Cabib, E (reprint author), NIDDK, Lab Biochem & Genet, NIH, Bldg 8,Rm 403, Bethesda, MD 20892 USA. NR 51 TC 199 Z9 206 U1 1 U2 8 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 8 PY 2001 VL 276 IS 23 BP 19679 EP 19682 DI 10.1074/jbc.R000031200 PG 4 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 439UC UT WOS:000169135100001 PM 11309404 ER PT J AU Urnov, FD Wolffe, AP AF Urnov, FD Wolffe, AP TI An array of positioned nucleosomes potentiates thyroid hormone receptor action in vivo SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID DNA-SEQUENCE SPECIFICITY; XENOPUS HSP70 PROMOTER; IN-VIVO; GLUCOCORTICOID RECEPTOR; TRANSCRIPTION FACTOR; MESSENGER-RNA; AMPHIBIAN METAMORPHOSIS; ANDROGEN RECEPTOR; RESPONSE ELEMENT; BETA GENE AB The assembly of the genome into chromatin imposes a poorly understood set of rules and constraints on action by regulatory factors. We investigated the role played by chromatin infrastructure in enabling an acute response of the Xenopus TR betaA gene to thyroid hormone receptor (TR), an extensively studied member of the nuclear hormone receptor superfamily, We found that in addition to the known TR response element (TRE) in the promoter, full range regulation required an upstream enhancer that contained multiple nonconsensus TREs and augmented ligand action at high receptor levels. An array of translationally positioned nucleosomes formed over the TR betaA locus in vivo; unliganded TR engaged this array in linker DNA between two nucleosomes and via TREs on the surface of histone octamers, Remarkably, assembly of enhancer DNA into mature chromatin potentiated binding by TR to its target response elements and enabled a greater range of regulation by TR than was observed on immature chromatin templates. Because assembly of enhancer DNA into chromatin increased TR binding to the nonconsensus TREs, we hypothesize that chromatin disruption targeted by liganded TR to the enhancer may lead to receptor release from the template and to an attenuation of response to hormone. C1 Sangamo Biosci, Point Richmond Tech Ctr, Richmond, CA 94804 USA. NICHD, Mol Embryol Lab, NIH, Bethesda, MD 20892 USA. RP Urnov, FD (reprint author), Sangamo Biosci, Point Richmond Tech Ctr, 501 Canal Blvd,Suite A100, Richmond, CA 94804 USA. NR 86 TC 15 Z9 15 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 8 PY 2001 VL 276 IS 23 BP 19753 EP 19761 DI 10.1074/jbc.M100924200 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 439UC UT WOS:000169135100012 PM 11274156 ER PT J AU Piek, E Ju, WJ Heyer, J Escalante-Alcalde, D Stewart, CL Weinstein, M Deng, CX Kucherlapati, R Bottinger, EP Roberts, AB AF Piek, E Ju, WJ Heyer, J Escalante-Alcalde, D Stewart, CL Weinstein, M Deng, CX Kucherlapati, R Bottinger, EP Roberts, AB TI Functional characterization of transforming growth factor beta signaling in Smad2-and Smad3-deficient fibroblasts SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID TGF-BETA; TARGETED DISRUPTION; IV COLLAGENASE; SMAD7 PROMOTER; CELL-LINES; FACTOR-SS; PATHWAY; GENE; ACTIVIN; BINDING AB A prominent pathway of transforming growth factor (TGF)-beta signaling involves receptor-dependent phosphorylation of Smad2 and Smad3, which then translocate to the nucleus to activate transcription of target genes. To investigate the relative importance of these two Smad proteins in TGF-beta1 signal transduction, we have utilized a loss of function approach, based on analysis of the effects of TGF-beta1 on fibroblasts derived from mouse embryos deficient in Smad2 (S2KO) or Smad3 (S3KO). TGF-beta1 caused 50% inhibition of cellular proliferation in wild-type fibroblasts as assessed by [H-3]thymidine incorporation, whereas the growth of S2KO or S3KO cells was only weakly inhibited by TGF-beta1. Lack of Smad2 or Smad3 expression did not affect TGF-1 beta -induced fibronectin synthesis but resulted in markedly suppressed induction of plasminogen activator inhibitor-1 by TGF-beta1. Moreover, TGF-beta1-mediated induction of matrix metalloproteinase-a was selectively dependent on Smad2, whereas induction of c-fos, Smad7, and TGF-beta1 autoinduction relied on expression of Smad3. Investigation of transcriptional activation of TGF-beta -sensitive reporter genes in the different fibroblasts showed that activation of the (Smad binding element)(4)-Lux reporter by TGF-beta1 was dependent on expression of Smad3, but not Smad2, whereas activation of the activin response element-Lux reporter was strongly suppressed in S2KO fibroblasts but, on the contrary, enhanced in S3KO cells. Our findings indicate specific roles for Smad2 and Smad3 in TGF-beta1 signaling. C1 NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bethesda, MD 20892 USA. Albert Einstein Coll Med, Dept Med, Bronx, NY 10461 USA. Albert Einstein Coll Med, Dept Mol Genet, Bronx, NY 10461 USA. NCI, Canc & Dev Biol Lab, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. NIDDK, Genet Dev & Dis Branch, NIH, Bethesda, MD 20892 USA. RP Piek, E (reprint author), NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bldg 41,Rm C629,41 Lib Dr,MSC-5055, Bethesda, MD 20892 USA. RI deng, chuxia/N-6713-2016 FU NIDDK NIH HHS [DK-56077-01] NR 45 TC 260 Z9 271 U1 0 U2 3 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 8 PY 2001 VL 276 IS 23 BP 19945 EP 19953 DI 10.1074/jbc.M102382200 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 439UC UT WOS:000169135100036 PM 11262418 ER PT J AU Yu, SH Castle, A Chen, M Lee, R Takeda, K Weinstein, LS AF Yu, SH Castle, A Chen, M Lee, R Takeda, K Weinstein, LS TI Increased insulin sensitivity in G(s)alpha knockout mice SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID TYROSINE KINASE-ACTIVITY; RAT ADIPOSE-CELLS; XL-ALPHA-S; TISSUE-SPECIFIC EXPRESSION; GLUCOSE-TRANSPORTER GLUT4; DEPENDENT PROTEIN-KINASE; HUMAN GNAS1 GENE; CYCLIC-AMP; SERINE/THREONINE PHOSPHORYLATION; SIGNAL-TRANSDUCTION AB The stimulatory guanine nucleotide-binding protein (G(s)) is required for hormone-stimulated cAMP generation. Gnas, the gene encoding the G(s) alpha -subunit, is imprinted, and targeted disruption of this gene in mice leads to distinct phenotypes in heterozygotes depending on whether the maternal (m-/f+) or paternal (+/p-) allele is mutated. Notably m-/+ mice become obese, whereas +/p- mice are thinner than normal. In this study we show that despite these opposite changes in energy metabolism, both m-/+ and +/p- mice have greater sensitivity to insulin, with low to normal fasting glucose levels, low fasting insulin levels, improved glucose tolerance, and exaggerated hypoglycemic response to administered insulin. The combination of increased insulin sensitivity with obesity in m-/+ mice is unusual, because obesity is typically associated with insulin resistance. In skeletal: muscles isolated hom both m-/+ and +/p- mice, the basal rate of 8-deoxyglucose uptake was normal, whereas the rate of 8-deoxyglucose uptake in response to maximal insulin stimulation was significantly increased. The similar changes in muscle sensitivity to insulin in m-/+ and +/p- mice may reflect the fact that muscle G(s)alpha expression is reduced by similar to 50% in both groups of mice; GLUT4 expression is unaffected in muscles from +/p- mice. Increased responsiveness to insulin is therefore the result of altered insulin signaling and/or GLUT4 translocation, This is the first direct demonstration in a genetically altered in vivo model that G(s)-coupled pathways negatively regulate insulin signaling. C1 NIDDK, Metab Dis Branch, NIH, Bethesda, MD 20892 USA. NIDDK, Diabet Branch, NIH, Bethesda, MD 20892 USA. RP Weinstein, LS (reprint author), NIDDK, Metab Dis Branch, NIH, Bldg 10,Rm 8C101, Bethesda, MD 20892 USA. NR 64 TC 49 Z9 49 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 8 PY 2001 VL 276 IS 23 BP 19994 EP 19998 DI 10.1074/jbc.M010313200 PG 5 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 439UC UT WOS:000169135100043 PM 11274197 ER PT J AU Luo, DL Broad, LM Bird, GS Putney, JW AF Luo, DL Broad, LM Bird, GS Putney, JW TI Mutual antagonism of calcium entry by capacitative and arachidonic acid-mediated calcium entry pathways SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID LACRIMAL ACINAR-CELLS; CA2+ ENTRY; INOSITOL TRISPHOSPHATE; CATION CHANNELS; HEK293 CELLS; RECEPTOR; RELEASE; MECHANISM; INHIBITION; ACTIVATION AB In nonexcitable cells, the predominant mechanism for regulated entry of Ca2+ is capacitative calcium entry, whereby depletion of intracellular Ca2+ stores signals the activation of plasma membrane calcium channels, A number of other regulated Ca2+ entry pathways occur in specific cell types, however, and it is not know to what degree the different pathways interact when present in the same cell. In this study, we have examined the interaction between capacitative calcium entry and arachidonic acid-activated calcium entry, which co-exist in HEK293 cells. These two pathways exhibit mutual antagonism. That is, capacitative calcium entry is potently inhibited by arachidonic acid, and arachidonic acid-activated entry is inhibited by the pre-activation of capacitative calcium entry with thapsigargin. In the latter case, the inhibition does not seem to result from a direct action of thapsigargin, inhibition of endoplasmic reticulum Ca2+ pumps, depletion of Ca2+ stores, or entry of Ca2+ through capacitative calcium entry channels. Rather, it seems that a discrete step in the pathway signaling capacitative calcium entry interacts with and inhibits the arachidonic acid pathway, The findings reveal a novel process of mutual antagonism between two distinct calcium entry pathways. This mutual antagonism may provide an important protective mechanism for the cell, guarding against toxic Ca2+ overload. C1 NIEHS, Lab Signal Transduct, NIH, Res Triangle Pk, NC 27709 USA. RP Putney, JW (reprint author), NIEHS, Lab Signal Transduct, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 30 TC 60 Z9 64 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 8 PY 2001 VL 276 IS 23 BP 20186 EP 20189 DI 10.1074/jbc.M100327200 PG 4 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 439UC UT WOS:000169135100070 PM 11274150 ER PT J AU Paria, BC Song, H Wang, X Schmid, PC Krebsbach, RJ Schmid, HHO Bonner, TI Zimmer, A Dey, SK AF Paria, BC Song, H Wang, X Schmid, PC Krebsbach, RJ Schmid, HHO Bonner, TI Zimmer, A Dey, SK TI Dysregulated cannabinoid signaling disrupts uterine receptivity for embryo implantation SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID PREIMPLANTATION MOUSE EMBRYO; LEUKEMIA INHIBITORY FACTOR; BLASTOCYST IMPLANTATION; CB1 RECEPTOR; UTERUS; EXPRESSION; ANANDAMIDE; PHARMACOLOGY; ACTIVATION; BINDS AB The mechanisms by which synchronized embryonic development to the blastocyst stage, preparation of the uterus for the receptive state, and reciprocal embryo-uterine interactions for implantation are coordinated are still unclear. We show in this study that preimplantation embryo development became asynchronous in mice that are deficient in brain-type (CB1) and/or spleen-type (CB2) cannabinoid receptor genes. Furthermore, whereas the levels of uterine anandamide (endocannabinoid) and blastocyst CB1 are coordinately down-regulated with the onset of uterine receptivity and blastocyst activation prior to implantation, these levels remained high in the nonreceptive uterus and in dormant blastocysts during delayed implantation and in pregnant, leukemia inhibitory factor (LIF)-deficient mice with implantation failure. These results suggest that a tight regulation of endocannabinoid signaling is important for synchronizing embryo development with uterine receptivity for implantation. Indeed this is consistent with our finding that while an experimentally induced, sustained level of an exogenously administered, natural cannabinoid inhibited implantation in wild-type mice, it failed to do so in CB1(-/-)/CB2(-/-) double mutant mice. The present study is clinically important because of the widely debated medicinal use of cannabinoids and their reported adverse effects on pregnancy. C1 Univ Kansas, Med Ctr, Ralph L Smith Res Ctr, Dept Mol & Intergrat Physiol, Kansas City, KS 66160 USA. Univ Kansas, Med Ctr, Dept Pediat, Kansas City, KS 66160 USA. Univ Minnesota, Hormel Inst, Austin, MN 55912 USA. NIMH, Genet Lab, Bethesda, MD 20892 USA. Univ Bonn, Psychiat Clin, Mol Neurobiol Lab, D-53105 Bonn, Germany. RP Dey, SK (reprint author), Univ Kansas, Med Ctr, Ralph L Smith Res Ctr, Dept Mol & Intergrat Physiol, 3901 Rainbow Blvd, Kansas City, KS 66160 USA. RI Zimmer, Andreas/B-8357-2009 FU NICHD NIH HHS [HD 02528, HD 1 2304, HD 29968, HD 33994, HD 37394]; NIDA NIH HHS [DA 06668]; NIGMS NIH HHS [GM 45741] NR 30 TC 119 Z9 124 U1 1 U2 2 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 8 PY 2001 VL 276 IS 23 BP 20523 EP 20528 DI 10.1074/jbc.M100679200 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 439UC UT WOS:000169135100114 PM 11279117 ER PT J AU Walters, RW Yi, SMP Keshavjee, S Brown, KE Welsh, MJ Chiorini, JA Zabner, J AF Walters, RW Yi, SMP Keshavjee, S Brown, KE Welsh, MJ Chiorini, JA Zabner, J TI Binding of adeno-associated virus type 5 to 2,3-linked sialic acid is required for gene transfer SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID RECOMBINANT ADENOASSOCIATED VIRUS; CILIATED AIRWAY EPITHELIA; INFLUENZA-C VIRUS; TRACHEAL EPITHELIUM; CANINE PARVOVIRUS; AAV VECTOR; FACTOR-IX; CELLS; RECEPTOR; TRANSDUCTION AB Recombinant adeno-associated viruses (AAV) are promising gene therapy vectors. Whereas AAV serotype 2-mediated gene transfer to muscle has partially replaced factor IX deficiency in hemophilia patients, its ability to mediate gene transfer to the lungs for cystic fibrosis is hindered by lack of apical receptors. However, AAV serotype 5 infects human airway epithelia from the lumenal surface. We found that in contrast to AAV2, the apical membrane of airway epithelia contains abundant high affinity receptors for AAV5. Binding and gene transfer with AAV5 was abolished by genetic or enzymatic removal of sialic acid from the cell surface. Furthermore, binding and gene transfer to airway epithelia was competed by lectins that specifically bind 2,3-linked sialic acid. These observations suggest that 2,3-linked sialic acid is either a receptor for AAV5 or it is a necessary component of a receptor complex. Further elucidation of the receptor for this virus should enhance understanding of parvovirus biology and expand the therapeutic targets for AAV vectors. C1 Univ Iowa, Coll Med, Dept Internal Med, Iowa City, IA 52242 USA. Univ Iowa, Coll Med, Dept Physiol & Biophys, Iowa City, IA 52242 USA. Univ Iowa, Coll Med, Dept Otolaryngol, Iowa City, IA 52242 USA. Univ Iowa, Coll Med, Howard Hughes Med Inst, Iowa City, IA 52242 USA. Univ Toronto, Toronto Lung Transplant Program, Toronto, ON M5G 2C4, Canada. NIDCR, Gene Therapeut Branch, NIH, Bethesda, MD 20892 USA. NHLBI, Hematol Branch, NIH, Bethesda, MD 20892 USA. RP Zabner, J (reprint author), Univ Iowa, Coll Med, Dept Internal Med, 500 EMRB, Iowa City, IA 52242 USA. FU NIDDK NIH HHS [T30DK54759] NR 61 TC 217 Z9 222 U1 0 U2 3 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 8 PY 2001 VL 276 IS 23 BP 20610 EP 20616 DI 10.1074/jbc.M101559200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 439UC UT WOS:000169135100126 PM 11262413 ER PT J AU Stunkel, W Ait-Si-Ali, S Jones, PL Wolffe, AP AF Stunkel, W Ait-Si-Ali, S Jones, PL Wolffe, AP TI Programming the transcriptional state of replicating methylated DNA SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID HISTONE DEACETYLASE; CPG ISLANDS; CHROMATIN; DEMETHYLATION; COMPLEX; EXPRESSION; BINDING; REPRESSION; ORIGINS; TARGET AB CpG methylation is maintained in daughter chromatids by the action of DNA methyltransferase at the replication fork. An opportunity exists for transcription factors at replication forks to bind their cognate sequences and thereby prevent remethylation by DNA methyltransferase. To test this hypothesis, we injected a linearized, methylated, and partially single-stranded re. porter plasmid into the nuclei of Xenopus oocytes and followed changes in the transcriptional activity after DNA replication. We find that dependent on Gal4-VP16, the action of DNA methyltransferase, and replication-coupled chromatin assembly DNA replication provides a window of time in which regulatory factors can activate or repress gene activity. Demethylation in the promoter region near the GAL4 binding sites of the newly synthesized DNA did not occur even though the Gal4 binding sites were occupied and transcription was activated. We conclude that "passive" demethylation at the replication fork is not simply dependent on the presence of DNA binding transcriptional activators. C1 NICHD, NIH, Bethesda, MD 20814 USA. RP Stunkel, W (reprint author), NICHD, NIH, Bldg 6,Rm 3A17, Bethesda, MD 20814 USA. NR 38 TC 3 Z9 3 U1 0 U2 0 PU AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0021-9258 J9 J BIOL CHEM JI J. Biol. Chem. PD JUN 8 PY 2001 VL 276 IS 23 BP 20743 EP 20749 DI 10.1074/jbc.M010967200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 439UC UT WOS:000169135100142 PM 11278800 ER PT J AU Young, SH Ye, JP Frazer, DG Shi, XL Castranova, V AF Young, SH Ye, JP Frazer, DG Shi, XL Castranova, V TI Molecular mechanism of tumor necrosis factor-alpha production in 1 -> 3-beta-glucan (Zymosan)-activated macrophages SO JOURNAL OF BIOLOGICAL CHEMISTRY LA English DT Article ID NF-KAPPA-B; POLYMORPHONUCLEAR LEUKOCYTES; TRANSCRIPTION FACTORS; GLUCAN RECEPTOR; TNF-ALPHA; ACTIVATION; CYTOKINE; CELLS; (1->3)-BETA-D-GLUCAN; STIMULATION AB The molecular details of 1 -->3-beta -glucans, a fungal cell wall component, induced inflammatory responses are not well understood. In the present study, we conducted a systematic analysis of the molecular events leading to tumor necrosis factor (TNF)-alpha production after glucan stimulation of macrophages. We demonstrated that activation of nuclear factor kappaB (NF-kappaB) is essential in zymosan A (a source of 1 -->3-beta -glucans)-induced TNF-alpha production in macrophages (RAW264.7 cells). Zymosan A-induced TNF-alpha protein production was associated with an increase in the TNF-alpha gene promoter activity. Activation of the TNF-alpha gene promoter was dependent on activation of NF-kappaB. Time course studies indicated that DNA binding activity of NF-kappaB preceded TNF-alpha promoter activity. Inhibition of NF-kappaB activation led to a dramatic reduction in both TNF-alpha promoter activity and TNF-alpha protein production in the response to zymosan A Mutation of a major NF-kappaB binding site (kappa3) in the gene promoter resulted in a significant decrease in the induction of the gene promoter by zymosan A, while mutation of Egr or CRE sites failed to inhibit the response to zymosan. Together, these results strongly suggest that NF-kappaB is involved in signal transduction of 1 -->3-beta -glucans-induced TNF-alpha expression. C1 NIOSH, ECTB, HELD, NIH, Morgantown, WV 26505 USA. NIOSH, Pathol & Physiol Res Branch, Hlth Effects Lab Div, NIH, Morgantown, WV 26505 USA. RP Young, SH (reprint author), NIOSH, ECTB, HELD, NIH, 1095 Willowdale Rd, Morgantown, WV 26505 USA. RI Shi, Xianglin/B-8588-2012 NR 41 TC 101 Z9 109 U1 0 U2 10 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 JUN 8 PY 2001 VL 276 IS 23 BP 20781 EP 20787 DI 10.1074/jbc.M101111200 PG 7 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 439UC UT WOS:000169135100147 PM 11259437 ER PT J AU Vuthoori, S Bowers, CW McCracken, A Dombroski, AJ Hinton, DM AF Vuthoori, S Bowers, CW McCracken, A Dombroski, AJ Hinton, DM TI Domain 1.1 of the sigma(70) subunit of Escherichia coli RNA polymerase modulates the formation of stable polymerase/promoter complexes SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE sigma(70); domain 1.1; domain 4.2; transcription; promoter ID BACTERIOPHAGE-T4 MIDDLE PROMOTER; TRANSCRIPTIONAL ACTIVATOR MOTA; SIGMA-70 SUBUNIT; BINDING-PROTEIN; DNA-BINDING; GENE; SPECIFICITY; INITIATION; SEQUENCES; MUTATION AB The sigma 70 (sigma (70)) subunit of Escherichia coli RNA polymerase specifies transcription from promoters that are responsible for basal gene expression during vegetative growth. When sigma (70) is present within polymerase holoenzyme, two of its domains, 2.4 and 4.2, interact with sequences within the -10 and -35 regions, respectively, of promoter DNA. However, in free sigma (70), DNA binding is prevented by domain 1.1, the N-terminal domain of the protein. Previous work has demonstrated that the presence of domain 1.1 is required for efficient transcription initiation at the lambda promoter P-R. To investigate whether this is a general property of domain 1.1, we have used five promoters to compare polymerases with and without domain 1.1 in in vitro transcription assays, and in assays assessing the formation and decay of stable, pretranscription complexes. We find that the absence of domain 1.1 does not render the polymerase defective at all of these promoters. Depending on the promoter, the absence of domain 1.1 can promote or inhibit transcription initiation by affecting the formation of stable pretranscription complexes. However, domain 1.1 does not affect the stability of these complexes once they are formed. For polymerases containing domain 1.1, the efficiency of stable complex formation correlates with how well the -10 and -35 regions of a promoter match the ideal sigma (70) recognition sequences. However, when domain 1.1 is absent, having this match becomes less important in determining how efficiently stable complexes are made. We suggest that domain 1.1 influences initiation by constraining polymerase to assess a promoter primarily by the fitness of its -10 and -35 regions to the canonical sequences. (C) 2001 Academic Press. C1 NIDDKD, Mol & Cellular Biol Lab, NIH, Bethesda, MD 20892 USA. Univ Texas, Hlth Sci Ctr, Dept Microbiol & Mol Genet, Houston, TX 77030 USA. RP Hinton, DM (reprint author), Regeneron Pharmaceut Inc, 777 Old Saw Mill River Rd, Princeton, NJ 08544 USA. NR 40 TC 49 Z9 50 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 JUN 8 PY 2001 VL 309 IS 3 BP 561 EP 572 DI 10.1006/jmbi.2001.4690 PG 12 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 442UG UT WOS:000169302900004 PM 11397080 ER PT J AU Kuznetsova, N McBride, DJ Leikin, S AF Kuznetsova, N McBride, DJ Leikin, S TI Osteogenesis imperfecta murine: Interaction between type I collagen homotrimers SO JOURNAL OF MOLECULAR BIOLOGY LA English DT Article DE collagen; fibrillogenesis; molecular interactions; osteogenesis imperfecta ID X-RAY-DIFFRACTION; TRIPLE HELICES; HYDRATION FORCES; FIBRIL STRUCTURE; VARIANT; MODEL; FIBRILLOGENESIS; MUTATION; PROTEINS; LAMPREY AB Types I, II, and III collagens are believed to have evolved from the same homotrimer ancestor and they have substantial sequence homology, but type I molecules are alpha1(I)(2)alpha2(I) heterotrimers, unlike homotrimeric types II and III. It is believed that the alpha2(I) chain first appeared in lower vertebrates formation. For instance, spontaneous mutations resulting in nonfunctional alpha2 chains and formation of type I homotrimers cause severe bone pathology (osteogenesis imperfecta) in humans and in animals. However, the exact role of the alpha2 chain is not known. Here, we report measurements of intermolecular forces between collagen helices in native and reconstituted fibers composed of type I homotrimers, heterotrimers and their mix. For comparison, we report forces between type II homotrimers in reconstituted fibers. In agreement with previous studies, we find that the absence of the alpha2 chain reduces temperature-favored attraction between collagen helices, either because of the difference in amino acid sequence of the alpha1 and alpha2 chains or because of more extensive post-translational modification of homotrimers. We find that forces between helices in fibers from type I las well as type Il) homotrimers are not sensitive to pH between pH 6 and 7.5, in contrast to type I heterotrimers. Apparently, the effect of pH is related to extra histidine residues present on alpha2 chains but not on alpha1 chains. Finally, our measurements indicate that the alpha2 chain is responsible for binding some soluble compound(s), possibly glycosaminoglycans, whose displacement results, e.g., in the loss of tendon crystallinity. The ability of the alpha2 chain to bind non-collagen matrix components may be particularly important for bone matrix formation and mineralization. (C) 2001 Academic Press. C1 NICHHD, Lab Phys & Struct Biol, NIH, Bethesda, MD 20892 USA. Univ Maryland, Sch Med, Div Endocrinol Diabet & Nutr, Baltimore, MD 21201 USA. RP Leikin, S (reprint author), NICHHD, Lab Phys & Struct Biol, NIH, Bethesda, MD 20892 USA. RI Leikin, Sergey/A-5518-2008 OI Leikin, Sergey/0000-0001-7095-0739 NR 31 TC 14 Z9 14 U1 0 U2 0 PU ACADEMIC PRESS LTD PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 0022-2836 J9 J MOL BIOL JI J. Mol. Biol. PD JUN 8 PY 2001 VL 309 IS 3 BP 807 EP 815 DI 10.1006/jmbi.2001.4682 PG 9 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 442UG UT WOS:000169302900022 PM 11397098 ER PT J AU Guo, XG Longnecker, MP Michalek, JE AF Guo, XG Longnecker, MP Michalek, JE TI Relation of serum tetrachlorodibenzo-p-dioxin concentration to diet among veterans in the Air Force Health Study with background-level exposure SO JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH-PART A LA English DT Article ID FOOD-FREQUENCY QUESTIONNAIRE; POLYCHLORINATED-BIPHENYLS; UNITED-STATES; REPRODUCIBILITY; DIBENZOFURANS; VALIDITY; SAMPLES; VIETNAM; PCBS; 2,3,7,8-TETRACHLORODIBENZO-P-DIOXIN AB To examine the contribution of various foods to exposure to 2,3,7,8-tetrachloro-p-dibenzodioxin (TCDD) in a background-exposed U.S. population, serum TCDD levels were examined in relation to diet as assessed by a standard diet assessment instrument among men with no known unusual exposure to TCDD. Our subjects were male veterans aged 42-76 yr who were in the unexposed comparison group in the Air Force Health Study, a study of the health effects of exposure to Agent Orange and herbicides in Vietnam. Food consumption was assessed by a 126-item food frequency questionnaire. Two hundred and ninety veterans who had both TCDD levels and diet assessed in 1992 were included. In general, associations between serum TCDD and consumption of foods in specific groups or nutrients, that is, meats and fats, were not evident. Among younger men, fish and chicken intake were associated with higher serum TCDD levels. The results suggest that no single rood group accounts for a large proportion of variation in TCDD exposure in older U.S. men. C1 NIEHS, Epidemiol Branch, Res Triangle Pk, NC 27709 USA. Analyt Sci Inc, Stat & Publ Hlth Res Div, Durham, NC USA. USAF, Res Lab, Brooks AFB, TX USA. RP Longnecker, MP (reprint author), NIEHS, Epidemiol Branch, POB 12233 MD A3-05, Res Triangle Pk, NC 27709 USA. OI Longnecker, Matthew/0000-0001-6073-5322 NR 33 TC 19 Z9 20 U1 0 U2 0 PU TAYLOR & FRANCIS LTD PI LONDON PA 11 NEW FETTER LANE, LONDON EC4P 4EE, ENGLAND SN 1528-7394 J9 J TOXICOL ENV HEAL A JI J. TOXICOL. ENV. HEALTH PT A PD JUN 8 PY 2001 VL 63 IS 3 BP 159 EP 172 DI 10.1080/15287390151101501 PG 14 WC Environmental Sciences; Public, Environmental & Occupational Health; Toxicology SC Environmental Sciences & Ecology; Public, Environmental & Occupational Health; Toxicology GA 439MU UT WOS:000169121300001 PM 11405413 ER PT J AU Varmus, H AF Varmus, H TI The growing family of NIH institutes - Response SO SCIENCE LA English DT Letter C1 Mem Sloan Kettering Canc Ctr, New York, NY 10021 USA. NIH, Bethesda, MD 20892 USA. RP Varmus, H (reprint author), Mem Sloan Kettering Canc Ctr, 1275 York Ave, New York, NY 10021 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 JUN 8 PY 2001 VL 292 IS 5523 BP 1836 EP 1836 PG 1 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 440WY UT WOS:000169200700019 ER PT J AU Nacro, K Sigano, DM Yan, SQ Nicklaus, MC Pearce, LL Lewin, NE Garfield, SH Blumberg, PM Marquez, VE AF Nacro, K Sigano, DM Yan, SQ Nicklaus, MC Pearce, LL Lewin, NE Garfield, SH Blumberg, PM Marquez, VE TI An optimized protein kinase C activating diacylglycerol combining high binding affinity (K-i) with reduced lipophilicity (log P) SO JOURNAL OF MEDICINAL CHEMISTRY LA English DT Article ID GREEN FLUORESCENT PROTEIN; PHORBOL ESTER; LIVING CELLS; DOMAINS; DELTA; DYNAMICS; CALCIUM; LIGANDS; DAG AB A small, focused combinatorial library encompassing all possible permutations of acyl branched alkyl chains-small and large, saturated and unsaturated-was generated from the active diacylglycerol enantiomer (S-DAG) to help identify the analogue with the highest binding affinity (lowest K-i) for protein kinase C (PK-C) combined with the minimum lipophilicity (log P). The selected ligand (3B) activated PK-C more effectively than sn-1,2-dioctanoylglycerol (diC8) despite being 1.4 log units more hydrophilic. Compound 3B indeed represents the most potent, hydrophilic DAG ligand to date. With the help of a green fluorescent protein (GFP)tagged PK-C alpha, 3B was able to translocate the full length protein to the membrane with an optimal dose of 100 muM in CHO-K1 cells, while diC8 failed to achieve translocation even at doses 3-fold higher. Molecular modeling of 3B into an empty C1b domain of PK-C delta clearly showed the existence of a preferred binding orientation. In addition, molecular dynamic simulations suggest that binding discrimination could result from a favorable van der Waals (VDW) interaction between the large, branched sn-1 acyl group of 3B and the aromatic rings of Trp252 (PK-C delta) or Tyr252 (PK-C alpha). The DAG analogue of 3B in which the acyl groups are reversed (2C) showed a decrease in binding affinity reflecting the capacity of PK-C to effectively discriminate between alternative orientations of the acyl chains. C1 NCI, Ctr Canc Res, Med Chem Lab, Frederick, MD 21702 USA. NCI, Ctr Canc Res, Cellular Carcinogenesis & Tumor Promot Lab, NIH, Bethesda, MD 20892 USA. NCI, Ctr Canc Res, Expt Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. RP Marquez, VE (reprint author), NCI, Ctr Canc Res, Med Chem Lab, Frederick, MD 21702 USA. RI Sigano, Dina/M-6144-2014; Nicklaus, Marc/N-4183-2014; OI Sigano, Dina/0000-0001-7489-9555; Nicklaus, Marc/0000-0002-4775-7030 NR 24 TC 25 Z9 25 U1 0 U2 0 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 JUN 7 PY 2001 VL 44 IS 12 BP 1892 EP 1904 DI 10.1021/jm010052e PG 13 WC Chemistry, Medicinal SC Pharmacology & Pharmacy GA 438HK UT WOS:000169047400006 PM 11384235 ER PT J AU Verrecchia, F Vindevoghel, L Lechleider, RJ Uitto, J Roberts, AB Mauviel, A AF Verrecchia, F Vindevoghel, L Lechleider, RJ Uitto, J Roberts, AB Mauviel, A TI Smad3/AP-1 interactions control transcriptional responses to TGF-beta in a promoter-specific manner SO ONCOGENE LA English DT Article DE TGF-beta; AP-1; Smad; gene regulation; Jun ID GROWTH-FACTOR-BETA; COLLAGEN GENE COL7A1; C-JUN; COACTIVATOR P300; SMAD PROTEINS; DNA-BINDING; ACTIVATION; FIBROBLASTS; EXPRESSION; SEQUENCE AB Smad proteins transduce signals from TGF-beta receptors and regulate transcription of target genes either directly or in combination with other sequence-specific transcription factors, AP-1 sites and their cognate transcription factors also play important roles in the gene regulatory activities of TGF-beta. In this report, we have investigated the functional interactions of the Smad and AP-1 transcription factors. We demonstrate that Smad and AP-1 complexes specifically bind to their cognate cis-elements and do not interact with each other on-DNA, whereas off-DNA interactions occur between Smad3 and both c-Jun and JunB. Using both artificial constructs specific for either the Smad or AP-1 signaling pathways or natural promoters known to be TGF-beta -responsive, we have determined that Jun family members downregulate Smad3-mediated gene transactivation whereas AP-1-dependent promoters are synergistically activated by Smad3 and Jun proteins, We propose a model where the presence of Smad- and/or AP-1-specific cis-elements within TGF-beta -responsive genes allows dynamic modulation of gene expression, in contrast to the existing model where interactions between Smad and AP-1 proteins are merely an on/off mechanism to regulate TGF-beta /Smad targets. C1 Hop St Louis, INSERM, U532, Inst Rech Peau, F-75010 Paris, France. Thomas Jefferson Univ, Jefferson Inst Mol Med, Dept Dermatol & Cutaneous Biol, Philadelphia, PA 19107 USA. NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bethesda, MD 20892 USA. RP Mauviel, A (reprint author), Hop St Louis, INSERM, U532, Inst Rech Peau, Pavillon Bazin,1 Ave Claude Vellefaux, F-75010 Paris, France. RI MAUVIEL, Alain/F-6251-2013 FU NIAMS NIH HHS [O1 AR41439] NR 38 TC 113 Z9 118 U1 2 U2 5 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD JUN 7 PY 2001 VL 20 IS 26 BP 3332 EP 3340 DI 10.1038/sj.onc.1204448 PG 9 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 441UW UT WOS:000169248800003 PM 11423983 ER PT J AU Minamoto, T Buschmann, T Habelhah, H Matusevich, E Tahara, H Boerresen-Dale, AL Harris, C Sidransky, D Ronai, Z AF Minamoto, T Buschmann, T Habelhah, H Matusevich, E Tahara, H Boerresen-Dale, AL Harris, C Sidransky, D Ronai, Z TI Distinct pattern of p53 phosphorylation in human tumors SO ONCOGENE LA English DT Article DE p53; phosphorylation; human tumor; JNK; ATM; p38 Chk ID DNA-DAMAGE; FUNCTION PHENOTYPE; CELL-CYCLE; IN-VITRO; KINASE; ACTIVATION; DOMAIN; BINDING; PROTEIN; MDM2 AB The protein product of the tumor suppressor gene p53 is phosphorylated on multiple residues by several protein kinases. Using a battery of 10 antibodies developed against different phosphorylated and acetylated residues of p53, we compared the pattern of p53 phosphorylation and acetylation in tumor-derived cell lines, tumor samples, and non-neoplastic cells. Irrespective of tumor types or the presence of p53 mutation, phosphorylation and acetylation of p53 was substantially higher in samples obtained from tumor tissues than those found in non-transformed samples. Among the 10 sites analysed, phosphorylation of residues 15, 81, 392, and acetylation were among the more frequent modifications. Analysis of two of the more abundant phosphorylation or acetylation sites on p53 is sufficient to detect 72% of tumor-derived p53 proteins. The distinct pattern of p53 phosphorylation and acetylation in human tumors may offer a new means to monitor the status and activity of p53 in the course of tumor development and progression. C1 Mt Sinai Sch Med, Ruttenberg Canc Ctr, New York, NY 10029 USA. Kanazawa Univ, Canc Res Inst, Kanazawa, Ishikawa 920, Japan. Hiroshima Univ, Sch Med, Dept Cellular & Mol Biol, Hiroshima, Japan. Norwegian Radium Hosp, Inst Canc Res, Dept Genet, Oslo, Norway. NCI, Human Carcinogenesis Lab, Bethesda, MD 20892 USA. Johns Hopkins Univ, Baltimore, MD USA. RP Ronai, Z (reprint author), Mt Sinai Sch Med, Ruttenberg Canc Ctr, 1 Gustave Levy Pl,Box 1130, New York, NY 10029 USA. OI RONAI, ZEEV/0000-0002-3859-0400 FU NCI NIH HHS [CA78419] NR 35 TC 60 Z9 64 U1 1 U2 4 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD JUN 7 PY 2001 VL 20 IS 26 BP 3341 EP 3347 DI 10.1038/sj.onc.1204458 PG 7 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 441UW UT WOS:000169248800004 PM 11423984 ER PT J AU Inga, A Resnick, MA AF Inga, A Resnick, MA TI Novel human p53 mutations that are toxic to yeast can enhance transactivation of specific promoters and reactivate tumor p53 mutants SO ONCOGENE LA English DT Article DE p53; yeast functional assays; p53 toxic mutants; enhanced transactivation; p53 mutants reactivation ID CELL-CYCLE ARREST; DNA-BINDING SITES; FUNCTIONAL ASSAY; GENE-EXPRESSION; SEQUENCE-SPECIFICITY; SUPPRESSOR PROTEIN; TERMINAL DOMAIN; GROWTH ARREST; CANCER-CELLS; TARGET GENE AB Since highly expressed human p53 can inhibit human and yeast cell growth, we predicted that p53 mutants could be generated with increased growth inhibition of the yeast Saccharomyces cerevisiae and that these would be useful for characterizing p53 functions and tumor p53 mutants. A random mutagenesis screen led to the isolation of mutations in the DIVA binding domain that result in p53 being lethal even at moderate expression levels in yeast. Three independent mutants had an alanine change at the evolutionary invariant V122 in the L1 loop. The other toxic mutations affected codons 277 (C277R, C277W) and 279 (G279R). This latter amino acid change was also reported in tumors, while all the other mutations are novel. A recently developed rheostatable GALI promoter system that provides graded increases in expression of p53 was used to examine the transactivation function of the toxic mutations when expression was greatly reduced and cells were viable. At low expression levels the toxic mutants lacked transactivation from a 3xRGC responsive element (RE). Surprisingly some exhibited enhanced transactivation with p21 and bax REs. The V122A mutant was able to re-activate transactivation of various p53 tumor mutants and retained growth inhibition when co-expressed with dominant-negative tumor mutations. Upon expression in human Saos-2 cells the V122A p53 mutant caused growth suppression, was capable of transactivation and exhibited higher than wild type activity with the bax promoter in luciferase assays. A non-functional p53 tumor mutant was partially reactivated by V122A for both transactivation and growth suppression. Thus, the screen for toxic p53 mutants in yeast can identify novel p53 variants that may be useful in dissecting p53 regulated cellular responses and in developing p53-based cancer therapies. C1 NIEHS, Genet Mol Lab, NIH, Res Triangle Pk, NC 27709 USA. RP Resnick, MA (reprint author), NIEHS, Genet Mol Lab, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 62 TC 32 Z9 33 U1 0 U2 0 PU NATURE PUBLISHING GROUP PI BASINGSTOKE PA HOUNDMILLS, BASINGSTOKE RG21 6XS, HAMPSHIRE, ENGLAND SN 0950-9232 J9 ONCOGENE JI Oncogene PD JUN 7 PY 2001 VL 20 IS 26 BP 3409 EP 3419 DI 10.1038/sj.onc.1204457 PG 11 WC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity SC Biochemistry & Molecular Biology; Oncology; Cell Biology; Genetics & Heredity GA 441UW UT WOS:000169248800011 PM 11423991 ER PT J AU Robey, RW Honjo, Y van de Laar, A Miyake, K Regis, JT Litman, T Bates, SE AF Robey, RW Honjo, Y van de Laar, A Miyake, K Regis, JT Litman, T Bates, SE TI A functional assay for detection of the mitoxantrone resistance protein, MXR (ABCG2) SO BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES LA English DT Article DE MXR/ABCP1/BCRP; mitoxantrone; adenosine 5 '-triphosphate-binding cassette transporter; drug resistance ID ACUTE MYELOID-LEUKEMIA; BREAST-CANCER CELLS; MULTIDRUG-RESISTANCE; P-GLYCOPROTEIN; HALF-TRANSPORTER; ATPASE ACTIVITY; DRUG-RESISTANCE; FUMITREMORGIN-C; GENE; OVEREXPRESSION AB The fluorescent compounds rhodamine 123, LysoTracker Green DMD-26, mitoxantrone, and BODIPY-prazosin were used with the antagonist fumitremorgin C (FTC) in order to develop functional assays for the half-transporter, MXR/BCRP/ABCP1. A measure of FTC-inhibitable efflux was generated for each compound in a series of MXR-overexpressing drug-selected cell lines and in ten unselected cell lines which were used to determine if the four fluorescent compounds were sensitive enough to detect the low MXR levels found in drug-sensitive cell lines. FTC-inhibitable efflux of mitoxantrone and prazosin was found in four of the ten cell lines, SF295, KM12, NCI-H460, and A549, and low but detectable levels of MXR mRNA were also observed by Northern analysis in these cells. FTC-inhibitable mitoxantrone and prazosin efflux in both selected and unselected cell lines was found to correlate well with MXR levels as determined by Northern blotting, r(2) = 0.89 and r(2) = 0.70 respectively. In contrast, rhodamine and LysoTracker were not able to reliably detect MXR. Cytotoxicity assays performed on two of the four unselected cell lines confirmed increased sensitivity to mitoxantrone in the presence of FTC. FTC was found to be a specific inhibitor of MXR, with half-maximal inhibition of MXR-associated ATPase activity at 1 muM FTC. Short term selections of the SF295, KM12, NCI-H460 and A549 cell lines in mitoxantrone resulted in a small but measurable increase in MXR by both Northern blot and functional assay. These studies show that flow cytometric measurement of FTC-inhibitable mitoxantrone or prazosin efflux is a sensitive and specific method for measuring the function of the MXR half-transporter in both selected and unselected cell lines. (C) 2001 Elsevier Science B.V. All rights reserved. C1 NCI, Dev Therapeut Dept, Med Branch, Ctr Canc Res,NIH, Bethesda, MD 20892 USA. RP Bates, SE (reprint author), NCI, Dev Therapeut Dept, Med Branch, Ctr Canc Res,NIH, Bldg 10,Room 12N226,9000 Rockville Pike, Bethesda, MD 20892 USA. NR 40 TC 174 Z9 182 U1 0 U2 5 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0005-2736 J9 BBA-BIOMEMBRANES JI Biochim. Biophys. Acta-Biomembr. PD JUN 6 PY 2001 VL 1512 IS 2 BP 171 EP 182 DI 10.1016/S0005-2736(01)00308-X PG 12 WC Biochemistry & Molecular Biology; Biophysics SC Biochemistry & Molecular Biology; Biophysics GA 444TG UT WOS:000169415700003 PM 11406094 ER PT J AU Agodoa, LY Appel, L Bakris, GL Beck, G Bourgoignie, J Briggs, JP Charleston, J Cheek, D Cleveland, W Douglas, JG Douglas, M Dowie, D Faulkner, M Gabriel, A Gassman, J Greene, T Hall, Y Hebert, L Hiremath, L Jamerson, K Johnson, CJ Kopple, J Kusek, J Lash, J Lea, J Lewis, JB Lipkowitz, M Massry, S Middleton, J Miller, ER Norris, K O'Connor, D Ojo, A Phillips, RA Pogue, V Rahman, M Randall, OS Rostand, S Schulman, G Smith, W Thornley-Brown, D Tisher, CC Toto, RD Wright, JT Xu, SC AF Agodoa, LY Appel, L Bakris, GL Beck, G Bourgoignie, J Briggs, JP Charleston, J Cheek, D Cleveland, W Douglas, JG Douglas, M Dowie, D Faulkner, M Gabriel, A Gassman, J Greene, T Hall, Y Hebert, L Hiremath, L Jamerson, K Johnson, CJ Kopple, J Kusek, J Lash, J Lea, J Lewis, JB Lipkowitz, M Massry, S Middleton, J Miller, ER Norris, K O'Connor, D Ojo, A Phillips, RA Pogue, V Rahman, M Randall, OS Rostand, S Schulman, G Smith, W Thornley-Brown, D Tisher, CC Toto, RD Wright, JT Xu, SC CA African Amer Study Kidney Dis Hype TI Effect of ramipril vs amlodipine on renal outcomes in hypertensive nephrosclerosis - A randomized controlled trial SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID CALCIUM-CHANNEL BLOCKERS; CONVERTING-ENZYME-INHIBITION; BLOOD-PRESSURE CONTROL; ISOLATED SYSTOLIC HYPERTENSION; DIABETIC NEPHROPATHY; AFRICAN-AMERICAN; NONDIABETIC NEPHROPATHIES; KIDNEY-DISEASE; ANTAGONISTS; PROGRESSION AB Context incidence of end-stage renal disease due to hypertension has increased in recent decades, but the optimal strategy for treatment of hypertension to prevent renal failure is unknown, especially among African Americans. Objective To compare the effects of an angiotensin-converting enzyme (ACE) inhibitor (ramipril), a dihydropyridine calcium channel blocker (amlodipine), and a beta -blocker (metoprolol) on hypertensive renal disease progression. Design, Setting, and Participants Interim analysis of a randomized, double-blind, 3 x 2 factorial trial conducted in 1094 African Americans aged 18 to 70 years with hypertensive renal disease (glomerular filtration rate [GFR] of 20-65 mL/min per 1.73 m(2)) enrolled between February. 1995 and September 1998. This report compares the ramipril and amlodipine groups following discontinuation of the amlodipine intervention in September 2000. Interventions Participants were randomly assigned to receive amlodipine, 15 to 10 mg/d (n=217), ramipril, 2.5 to 10 mg/d (n=436), or metoprolol, 50 to 200 mg/d (n=441), with other agents added to achieve 1 of 2 blood pressure goals. Main Outcome Measures The primary outcome measure was the rate of change in GFR; the main secondary outcome was a composite index of the clinical end points of reduction in GFR of more than 50% or 25 mL/min per 1.73 m(2), end-stage renal disease, or death. Results Among participants with a urinary protein to creatinine ratio of >0.22 (corresponding approximately to proteinuria of more than 300 mg/d), the ramipril group had a 36% (2.02 [SE, 0.74] mL/min per 1.73 m(2)/y) slower mean decline in GFR over 3 years (P=.006) and a 48% reduced risk of the clinical end points vs the amlodipine group (95% confidence interval [CI], 20%-66%). In the entire cohort, there was no significant difference in mean GFR decline from baseline to 3 years between treatment groups (P=.38). However,compared with the amlodipine group, after adjustment for baseline covariates the ramipril group had a 38% reduced risk of clinical end points (95% CI, 13% -56%), a 36% slower mean decline in GFR after 3 months (P=.002), and less proteinuria (P<.001). Conclusion Ramipril, compared with amlodipine, retards renal disease progression in patients with hypertensive renal disease and proteinuria and may offer benefit to patients without proteinuria. C1 Louis Stokes, Cleveland VA Med Ctr, Cleveland, OH 44106 USA. NIDDKD, Bethesda, MD 20892 USA. Johns Hopkins Univ, Dept Prevent Med, Baltimore, MD USA. Rush Presbyterian St Lukes Med Ctr, Dept Prevent Med, Chicago, IL 60612 USA. Cleveland Clin Fdn, Dept Biostat, Cleveland, OH 44195 USA. Univ Miami, Dept Med, Coral Gables, FL 33124 USA. Med Univ S Carolina, Dept Med, Charleston, SC 29425 USA. Morehouse Sch Med, Dept Med, Atlanta, GA 30310 USA. Case Western Reserve Univ Hosp, Dept Med, Cleveland, OH 44106 USA. Emory Univ, Med Ctr, Dept Med, Atlanta, GA 30322 USA. Harlem Hosp Ctr, Dept Med, New York, NY USA. Meharry Med Coll, Dept Med, Nashville, TN 37208 USA. Mt Sinai Sch Med, Dept Med, New York, NY USA. Ohio State Univ, Dept Med, Columbus, OH 43210 USA. Univ Michigan, Dept Med, Ann Arbor, MI 48109 USA. Univ Calif Los Angeles, Harbor Med Ctr, Dept Med, Los Angeles, CA 90024 USA. Univ Illinois, Dept Med, Chicago, IL USA. Vanderbilt Univ, Dept Med, Nashville, TN USA. Univ So Calif, Dept Med, Los Angeles, CA USA. Univ Texas, SW Med Ctr, Dept Med, Dallas, TX USA. Martin L King Charles R Drew Med Ctr, Dept Med, Los Angeles, CA USA. Univ Calif San Diego, Dept Med, San Diego, CA 92103 USA. Howard Univ, Dept Med, Washington, DC 20059 USA. Univ Alabama, Dept Med, Birmingham, AL 35294 USA. Univ Florida, Dept Med, Gainesville, FL USA. RP Wright, JT (reprint author), Case Western Reserve Univ, Univ Hosp Cleveland, Clin Hypertens Program, 10900 Euclid Ave,Wood Bldg Room W-165, Cleveland, OH 44106 USA. RI Briggs, Josephine/B-9394-2009 OI Briggs, Josephine/0000-0003-0798-1190 FU NCRR NIH HHS [M01 RR-00080, 2P20 RR11104, 5M01 RR-00071, M01 RR00827, P20-RR11145]; PHS HHS [M01 00032] NR 40 TC 549 Z9 569 U1 3 U2 28 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 JUN 6 PY 2001 VL 285 IS 21 BP 2719 EP 2728 DI 10.1001/jama.285.21.2719 PG 10 WC Medicine, General & Internal SC General & Internal Medicine GA 437HG UT WOS:000168985800024 PM 11386927 ER PT J AU Schiffmann, R Kopp, JB Austin, HA Sabnis, S Moore, DF Weibel, T Balow, JE Brady, RO AF Schiffmann, R Kopp, JB Austin, HA Sabnis, S Moore, DF Weibel, T Balow, JE Brady, RO TI Enzyme replacement therapy in Fabry disease - A randomized controlled trial SO JAMA-JOURNAL OF THE AMERICAN MEDICAL ASSOCIATION LA English DT Article ID CANCER PAIN; MANIFESTATIONS; INVOLVEMENT; SEVERITY; STORAGE AB Context Fabry disease is a metabolic disorder without a specific treatment, caused by a deficiency of the lysosomal enzyme alpha -galactosidase A (alpha -gal A). Most patients experience debilitating neuropathic pain and premature mortality because of renal failure, cardiovascular disease, or cerebrovascular disease. Objective To evaluate the safety and efficacy of intravenous alpha -gal A for Fabry disease. Design and Setting Double-blind placebo-controlled trial conducted from December 1998 to August 1999 at the Clinical Research Center of the National Institutes of Health. Patients Twenty-six hemizygous male patients, aged 18 years or older, with Fabry disease that was confirmed by alpha -gal A assay, Intervention A dosage of 0.2 mg/kg of alpha -gal A, administered intravenously every other week (12 doses total), Main Outcome Measure Effect of therapy on neuropathic pain while without neuropathic pain medications measured by question 3 of the Brief Pain Inventory (BPI), Results Mean (SE) BPI neuropathic pain severity score declined from 6.2 (0.46) to 4.3 (0.73) in patients treated with alpha -gal A vs no significant change in the placebo group (P =.02). Pain-related quality of life declined from 3.2 (0.55) to 2.1 (0.56) for patients receiving alpha -gal A vs 4.8 (0.59) to 4.2 (0.74) for placebo (P =.05). In the kidney, glomeruli with mesangial widening decreased by a mean of 12.5% for patients receiving alpha -gal vs a 16.5% increase for placebo (P =.01), Mean inulin clearance decreased by 6.2 mL/min for patients receiving alpha -gal A vs 19.5 mL/min for placebo (P =.19). Mean creatinine clearance increased by 2.1 mL/min (0.4 mL/s) for patients receiving a-gal A vs a decrease of 16.1 mL/min (0.3 mL/s) for placebo (P =.02). In patients treated with alpha -gal A, there was an approximately 50% reduction in plasma glycosphingolipid levels, a significant improvement in cardiac conduction, and a significant increase in body weight. Conclusion Intravenous infusions of alpha -gal A are safe and have widespread therapeutic efficacy in Fabry disease. C1 NINDS, Dev & Metab Neurol Branch, NIH, Bethesda, MD 20892 USA. NIDDKD, Kidney Dis Sect, NIH, Bethesda, MD 20892 USA. Armed Forces Inst Pathol, Div Nephropathol, Washington, DC 20306 USA. RP Schiffmann, R (reprint author), NINDS, Dev & Metab Neurol Branch, NIH, Bldg 10,Room 3D03,9000 Rockville Pike, Bethesda, MD 20892 USA. OI Kopp, Jeffrey/0000-0001-9052-186X NR 30 TC 768 Z9 800 U1 2 U2 21 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 JUN 6 PY 2001 VL 285 IS 21 BP 2743 EP 2749 DI 10.1001/jama.285.21.2743 PG 7 WC Medicine, General & Internal SC General & Internal Medicine GA 437HG UT WOS:000168985800027 PM 11386930 ER PT J AU Howe, HL Wingo, PA Thun, MJ Ries, LAG Rosenberg, HM Feigal, EG Edwards, BK AF Howe, HL Wingo, PA Thun, MJ Ries, LAG Rosenberg, HM Feigal, EG Edwards, BK TI Annual report to the nation on the status of cancer (1973 through 1998), featuring cancers with recent increasing trends SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Review ID PAPILLARY THYROID-CANCER; SOFT-TISSUE SARCOMA; CHRONIC HEPATITIS-B; BREAST-CANCER; UNITED-STATES; PROSTATE-CANCER; SURVEILLANCE SERIES; INTERPRETING TRENDS; COLORECTAL-CANCER; GASTRIC CARDIA AB Background: The American Cancer Society, the National Cancer Institute (NCI), the North American Association of Central Cancer Registries, and the Centers for Disease Control and Prevention, including the National Center for Health Statistics (NCHS), collaborate to provide an annual update on cancer occurrence and trends in the United States. This year's report contains a special feature that focuses on cancers with recent increasing trends. Methods: From 1992 through 1998, age-adjusted rates and annual percent changes are calculated for cancer incidence and underlying cause of death with the use of NCI incidence and NCHS mortality data. Joinpoint analysis, a model of joined line segments, is used to examine long-term trends for the four most common cancers and for those cancers with recent increasing trends in incidence or mortality. Statistically significant findings are based on a P value of .05 by use of a two-sided test. State-specific incidence and death rates for 1994 through 1998 are reported for major cancers. Results: From 1992 through 1998, total cancer death rates declined in males and females, while cancer incidence rates declined only in males, Incidence rates in females increased slightly, largely because of breast cancer increases that occurred in some older age groups, possibly as a result of increased early detection. Female lung cancer mortality, a major cause of death in women, continued to increase but more slowly than in earlier years. In addition, the incidence or mortality rate increased in 10 other sites, accounting for about 13% of total cancer incidence and mortality in the United States. Conclusions: Overall cancer incidence and death rates continued to decline in the United States, Future progress will require sustained improvements in cancer prevention, screening, and treatment. C1 N Amer Assoc Cent Canc Registries Inc, Springfield, IL 62704 USA. Ctr Dis Control & Prevent, Natl Ctr Chron Dis Prevent & Hlth Promot, Div Canc Prevent & Control, Atlanta, GA USA. Amer Canc Soc, Epidemiol & Surveillance Res Dept, Atlanta, GA 30329 USA. NCI, Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. NCI, Div Canc Treatment & Diag, Bethesda, MD 20892 USA. Natl Ctr Hlth Stat, Div Vital Stat, Hyattsville, MD 20782 USA. RP Howe, HL (reprint author), N Amer Assoc Cent Canc Registries Inc, 2121 W White Oaks Dr, Springfield, IL 62704 USA. NR 107 TC 521 Z9 541 U1 0 U2 8 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 JUN 6 PY 2001 VL 93 IS 11 BP 824 EP 842 DI 10.1093/jnci/93.11.824 PG 19 WC Oncology SC Oncology GA 439HH UT WOS:000169105100009 PM 11390532 ER PT J AU Dong, SM Traverso, G Johnson, C Geng, L Favis, R Boynton, K Hibi, K Goodman, SN D'Allessio, M Paty, P Hamilton, SR Sidransky, D Barany, F Levin, B Shuber, A Kinzler, KW Vogelstein, B Jen, J AF Dong, SM Traverso, G Johnson, C Geng, L Favis, R Boynton, K Hibi, K Goodman, SN D'Allessio, M Paty, P Hamilton, SR Sidransky, D Barany, F Levin, B Shuber, A Kinzler, KW Vogelstein, B Jen, J TI Detecting colorectal cancer in stool with the use of multiple genetic targets SO JOURNAL OF THE NATIONAL CANCER INSTITUTE LA English DT Article ID K-RAS MUTATIONS; MICROSATELLITE INSTABILITY; DNA-LIGASE; P53 GENE; TUMORS; PCR; DATABASE; GERMLINE; FIDELITY; ADENOMAS AB Background: Colorectal cancer cells are shed into the stool, providing a potential means for the early detection of the disease using noninvasive approaches. Our goal was to develop reliable, specific molecular genetic tests for the detection of colorectal cancer in stool samples. Methods: Stool DNA was isolated from paired stools and primary tumor samples from 51 colorectal cancer patients. Three genetic targets-TP53, BAT26, and K-RAS-were used to detect tumor-associated mutations in the stool prior to or without regard to the molecular analyses of the paired tumors. TP53 gene mutations were detected with a mismatch-ligation assay that detects nine common p53 gene mutations. Deletions within the BAT26 locus were detected by a modified solid-phase minisequencing method. Mutations in codons 12 and 13 of K-RAS were detected with a digital polymerase chain reaction-based method. Results: TP53 gene mutations were detected in the tumor DNA of 30 patients, all of whom had the identical TP53 mutation in their stools. Tumors from three patients contained a noninherited deletion at the BAT26 locus, and the same alterations were identified in these patients' stool specimens. Nineteen of 50 tumors tested had a KRAS mutation; identical mutations were detected in the paired stool DNA samples from eight patients. In no case was a mutation found in stool that was not also present in the primary tumor. Thus, the three genetic markers together detected 36 (71%) of 51 patients (95 % confidence interval [CI] = 56% to 83%) with colorectal cancer and 36 (92%) of 39 patients (95% CI = 79% to 98%) whose tumors had an alteration. Conclusion: We were able to detect the majority of colorectal cancers by analyzing stool DNA for just three genetic markers. Additional work is needed to determine the specificity of these genetic tests for detecting colorectal neoplasia in asymptomatic patients and to more precisely estimate the prevalence of the mutations and sensitivity of the assay. C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Johns Hopkins Med Sch, Johns Hopkins Oncol Ctr, Dept Otolaryngol Head & Neck Surg, Div Head & Neck Canc Res, Bethesda, MD USA. EXACT Labs Inc, Maynard, MA USA. Mem Sloan Kettering Canc Ctr, New York, NY 10021 USA. Cornell Univ, Dept Microbiol, Ithaca, NY USA. Univ Texas, MD Anderson Canc Ctr, Houston, TX USA. Johns Hopkins Univ, Johns Hopkins Oncol Ctr, Baltimore, MD USA. Johns Hopkins Univ, Howard Hughes Med Inst, Baltimore, MD USA. Johns Hopkins Univ, Program Human Genet, Baltimore, MD USA. Johns Hopkins Univ, Johns Hopkins Med Sch, Dept Otolaryngol, Div Head & Neck Res, Baltimore, MD USA. RP Jen, J (reprint author), NIH, Bldg 41,Rm D702,41 Lib Dr, Bethesda, MD 20892 USA. FU NCI NIH HHS [CA62924, CA65930]; NIGMS NIH HHS [GM07184] NR 31 TC 256 Z9 269 U1 2 U2 8 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 JUN 6 PY 2001 VL 93 IS 11 BP 858 EP 865 DI 10.1093/jnci/93.11.858 PG 8 WC Oncology SC Oncology GA 439HH UT WOS:000169105100012 PM 11390535 ER PT J AU Deerfield, DW Carter, CW Pedersen, LG AF Deerfield, DW Carter, CW Pedersen, LG TI Models for protein-zinc ion binding sites. II. The catalytic sites SO INTERNATIONAL JOURNAL OF QUANTUM CHEMISTRY LA English DT Article DE zinc; catalytic sites; ab initio; ligands; coordination ID TRANSITION-STATE ANALOG; QUANTUM-CHEMICAL CALCULATIONS; DENSITY-FUNCTIONAL THEORY; CARBONIC ANHYDRASE-II; VALENCE SUM ANALYSIS; LIGAND BOND LENGTHS; AB-INITIO; CYTIDINE DEAMINASE; ALCOHOL-DEHYDROGENASE; CRYSTAL-STRUCTURE AB The first and selected members of the second coordination shells for four model protein-Zn(II) ion catalytic sites have been studied using ab initio computational methodology. The influence of deprotonation on the structure and relative energetics of model complexes was examined. Significant lengthening of the Zn(II)-S, Zn(II)-O, and Zn(II)-N ionic distance is observed as the negative charge about the Zn(II) ion increases. In a model Escherichia coli cytidine deaminase site, we find a substantial lengthening of the Zn-SH2 ionic distance in the active site, which corresponds to the lengthening found for transition state analogs in X-ray crystal structures. The lowest energy complex of a sulfur-containing ligand was found to be singly deprotonated, with the complex having an overall charge of +1. The deprotonation of any ligand in a model complex for thermolysin, on the other hand. was found to be endothermic. The influence of other ligands of the metal ion, along with second-shell ligands, on the energy requirement for deprotonation M as also examined for other model systems. (C) 2001 John Wiley & Sons. Inc. C1 Carnegie Mellon Univ, Pittsburgh Supercomp Ctr, Pittsburgh, PA 15213 USA. Univ N Carolina, Dept Biochem & Biophys, Chapel Hill, NC 27599 USA. Univ N Carolina, Dept Chem, Chapel Hill, NC 27599 USA. NIEHS, Res Triangle Pk, NC 27709 USA. RP Deerfield, DW (reprint author), Carnegie Mellon Univ, Pittsburgh Supercomp Ctr, Pittsburgh, PA 15213 USA. RI Pedersen, Lee/E-3405-2013 OI Pedersen, Lee/0000-0003-1262-9861 NR 85 TC 30 Z9 30 U1 0 U2 4 PU JOHN WILEY & SONS INC PI NEW YORK PA 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0020-7608 J9 INT J QUANTUM CHEM JI Int. J. Quantum Chem. PD JUN 5 PY 2001 VL 83 IS 3-4 BP 150 EP 165 DI 10.1002/qua.1207 PG 16 WC Chemistry, Physical; Mathematics, Interdisciplinary Applications; Physics, Atomic, Molecular & Chemical SC Chemistry; Mathematics; Physics GA 430QT UT WOS:000168587600006 ER PT J AU Horkay, F Hecht, AM Basser, PJ Geissler, E AF Horkay, F Hecht, AM Basser, PJ Geissler, E TI Comparison between neutral gels and neutralized polyelectrolyte gels in the presence of divalent cations SO MACROMOLECULES LA English DT Article ID SCATTERING C1 NIH, Sect Tissue Biophys & Biomimet, Lab Integrat & Med Biophys, Bethesda, MD 20892 USA. Univ Grenoble 1, Lab Spectrometrie Phys, CNRS UMR 5588, F-38402 St Martin Dheres, France. RP Horkay, F (reprint author), NIH, Sect Tissue Biophys & Biomimet, Lab Integrat & Med Biophys, Bldg 10, Bethesda, MD 20892 USA. RI Basser, Peter/H-5477-2011 NR 18 TC 10 Z9 10 U1 1 U2 9 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0024-9297 J9 MACROMOLECULES JI Macromolecules PD JUN 5 PY 2001 VL 34 IS 12 BP 4285 EP 4287 DI 10.1021/ma0021850 PG 3 WC Polymer Science SC Polymer Science GA 438JC UT WOS:000169049000068 ER PT J AU Tang, PZ Tsai-Morris, CH Dufau, ML AF Tang, PZ Tsai-Morris, CH Dufau, ML TI Cloning and characterization of a hormonally regulated rat long chain acyl-CoA synthetase SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID COMPLETE GENOME SEQUENCE; LEYDIG-CELLS; GONADOTROPIN; EXPRESSION; PURIFICATION; MODULATION; CDNA AB A previously unidentified gonadotropin-regulated long chain acyl-CoA synthetase (GR-LACS) was cloned and characterized as a 79-kDa cytoplasmic protein expressed in Leydig cells of the rat testis, GR-LACS shares sequence identity with two conserved regions of the LACS and luciferase families, including the ATP/AMP binding domain and the 25-aa fatty acyl-CoA synthetase signature motif, but displays low overall amino acid similarities (23-28%), GR-LACS mRNA is expressed abundantly in Leydig cells of the adult testis and to a lesser degree in the seminiferous tubules in spermatogonia and Sertoli cells. It is also observed in ovary and brain. Immunoreactive protein expression was observed mainly in Leydig cells and minimally in the tubules but was not detected in other tissues, In vivo, treatment with a desensitizing dose of human chorionic gonadotropin caused transcriptional down-regulation of CR-LACS expression in Leydig cells. The expressed protein present in the cytoplasm of transfected cells displayed acyl-CoA synthetase activity for long chain fatty acid substrates. CR-LACS may contribute to the provision of energy requirements and to the biosynthesis of steroid precursors and could participate through acyl-CoA's multiple functions in the regulation of the male gonad. C1 NICHHD, Sect Mol Endocrinol, Endocrinol & Reprod Res Branch, NIH, Bethesda, MD 20892 USA. RP Dufau, ML (reprint author), NICHHD, Sect Mol Endocrinol, Endocrinol & Reprod Res Branch, NIH, Bldg 49,Room 6A36, Bethesda, MD 20892 USA. NR 26 TC 22 Z9 25 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 JUN 5 PY 2001 VL 98 IS 12 BP 6581 EP 6586 DI 10.1073/pnas.121046998 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 440AT UT WOS:000169151500012 PM 11381125 ER PT J AU Zavadil, J Bitzer, M Liang, D Yang, YC Massimi, A Kneitz, S Piek, E Bottinger, EP AF Zavadil, J Bitzer, M Liang, D Yang, YC Massimi, A Kneitz, S Piek, E Bottinger, EP TI Genetic programs of epithelial cell plasticity directed by transforming growth factor-beta SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID TGF-BETA; MESENCHYMAL TRANSITION; TRANSGENIC MICE; SMAD PROTEINS; CYCLE ARREST; II RECEPTOR; EXPRESSION; PATHWAY; REVEALS; TRANSDIFFERENTIATION AB Epithelial-mesenchymal transitions (EMTs) are an essential manifestation of epithelial cell plasticity during morphogenesis, wound healing, and tumor progression. Transforming growth factor-beta (TGF-beta) modulates epithelial plasticity in these physiological contexts by inducing EMT. Here we report a transcriptome screen of genetic programs of TGF-beta -induced EMT in human keratinocytes and propose functional roles for extracellular response kinase (ERK) mitogen-activated protein kinase signaling in cell motility and disruption of adherens junctions. We used DNA arrays of 16,580 human cDNAs to identify 728 known genes regulated by TGF-beta within 4 hours after treatment. TGF-beta -stimulated ERK signaling mediated regulation of 80 target genes not previously associated with this pathway. This subset is enriched for genes with defined roles in cell-matrix interactions, cell motility, and endocytosis. ERK-independent genetic programs underlying the onset of EMT involve key pathways and regulators of epithelial dedifferentiation, undifferentiated transitional and mesenchymal progenitor phenotypes, and mediators of cytoskeletal reorganization. The gene expression profiling approach delineates complex context-dependent signaling pathways and transcriptional events that determine epithelial cell plasticity controlled by TGF-beta. Investigation of the identified pathways and genes will advance the understanding of molecular mechanisms that underlie tumor invasiveness and metastasis. C1 Yeshiva Univ Albert Einstein Coll Med, Dept Med, Bronx, NY 10461 USA. Yeshiva Univ Albert Einstein Coll Med, Dept Mol Genet, Bronx, NY 10461 USA. NCI, Lab Cell Regulat & Carcinogenesis, NIH, Bethesda, MD 20892 USA. RP Bottinger, EP (reprint author), Yeshiva Univ Albert Einstein Coll Med, Dept Med, 1300 Morris Pk Ave, Bronx, NY 10461 USA. OI Zavadil, Jiri/0000-0003-0640-5562 FU NIDDK NIH HHS [R01 DK056077] NR 44 TC 350 Z9 365 U1 2 U2 11 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 JUN 5 PY 2001 VL 98 IS 12 BP 6686 EP 6691 DI 10.1073/pnas.111614398 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 440AT UT WOS:000169151500030 PM 11390996 ER PT J AU Ellis, HM Yu, DG DiTizio, T Court, DL AF Ellis, HM Yu, DG DiTizio, T Court, DL TI High efficiency mutagenesis, repair, and engineering of chromosomal DNA using single-stranded oligonucleotides SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID COLI RECBCD ENZYME; ESCHERICHIA-COLI; BETA-PROTEIN; PHAGE-LAMBDA; BACTERIOPHAGE-LAMBDA; SYNTHETIC OLIGONUCLEOTIDES; RECOMBINATION SYSTEM; TRANSFORMATION; YEAST; GENE AB Homologous DNA recombination is a fundamental, regenerative process within living organisms. However, in most organisms, homologous recombination is a rare event, requiring a complex set of reactions and extensive homology, We demonstrate in this paper that Beta protein of phage A generates recombinants in chromosomal DNA by using synthetic single-stranded DNAs (ssDNA) as short as 30 bases long. This ssDNA recombination can be used to mutagenize or repair the chromosome with efficiencies that generate up to 6% recombinants among treated cells. Mechanistically, it appears that Beta protein, a Rad52-like protein, binds and anneals the ssDNA donor to a complementary single-strand near the DNA replication fork to generate the recombinant, This type of homologous recombination with ssDNA provides new avenues for studying and modifying genomes ranging from bacterial pathogens to eukaryotes, Beta protein and ssDNA may prove generally applicable for repairing DNA in many organisms. C1 NCI, Gene Regulat & Chromosome Biol Lab, Div Basic Sci, Ft Detrick, MD 21702 USA. RP Court, DL (reprint author), NCI, Gene Regulat & Chromosome Biol Lab, Div Basic Sci, Bldg 539,Room 243,POB B, Ft Detrick, MD 21702 USA. NR 28 TC 272 Z9 296 U1 5 U2 48 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 JUN 5 PY 2001 VL 98 IS 12 BP 6742 EP 6746 DI 10.1073/pnas.121164898 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 440AT UT WOS:000169151500040 PM 11381128 ER PT J AU Webster, JC Oakley, RH Jewell, CM Cidlowski, JA AF Webster, JC Oakley, RH Jewell, CM Cidlowski, JA TI Proinflammatory cytokines regulate human glucocorticoid receptor gene expression and lead to the accumulation of the dominant negative beta isoform: A mechanism for the generation of glucocorticoid resistance SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID NF-KAPPA-B; SYSTEMIC LUPUS-ERYTHEMATOSUS; ALZHEIMERS-DISEASE; CROSS-TALK; IMMUNOCYTOCHEMICAL ANALYSIS; ANTIINFLAMMATORY DRUGS; RHEUMATOID-ARTHRITIS; DOWN-REGULATION; ASTHMA; DEXAMETHASONE AB Inflammatory responses in many cell types are coordinately regulated by the opposing actions of NF-kappaB and the glucocorticoid receptor (GR), The human glucocorticoid receptor (hGR) gene encodes two protein isoforms: a cytoplasmic alpha form (GR alpha), which binds hormone, translocates to the nucleus, and regulates gene transcription, and a nuclear localized beta isoform (GRP), which does not bind known ligands and attenuates GR alpha action. We report here the identification of a tumor necrosis factor (TNF)responsive NF-kappaB DNA binding site 5' to the hCR promoter that leads to a 1.5-fold increase in GR alpha mRNA and a 2.0-fold increase in GRP mRNA in HeLa53 cells, which endogenously express both GR isoforms, However, TNF-alpha treatment disproportionately increased the steady-state levels of the GR beta protein isoform over GR alpha, making GR beta the predominant endogenous receptor isoform, Similar results were observed following treatment of human CEMC7 lymphoid cells with TNF-alpha or IL-1, The increase in GRP protein expression correlated with the development of glucocorticoid resistance. C1 NIEHS, Lab Signal Transduct, Mol Endocrinol Grp, NIH, Res Triangle Pk, NC 27709 USA. RP Cidlowski, JA (reprint author), NIEHS, Lab Signal Transduct, Mol Endocrinol Grp, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 54 TC 282 Z9 311 U1 5 U2 10 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 JUN 5 PY 2001 VL 98 IS 12 BP 6865 EP 6870 DI 10.1073/pnas.121455098 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 440AT UT WOS:000169151500061 PM 11381138 ER PT J AU Weise, M De-Levi, S Barnes, KM Gafni, RI Abad, V Baron, J AF Weise, M De-Levi, S Barnes, KM Gafni, RI Abad, V Baron, J TI Effects of estrogen on growth plate senescence and epiphyseal fusion SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID ZEALAND WHITE-RABBIT; LONGITUDINAL BONE-GROWTH; AROMATASE DEFICIENCY; PRECOCIOUS PUBERTY; SKELETAL TISSUES; CELL-KINETICS; GUINEA-PIGS; LONG BONES; RATS; HORMONE AB Estrogen is critical for epiphyseal fusion in both young men and women. In this study, we explored the cellular mechanisms by which estrogen causes this phenomenon. Juvenile ovariectomized female rabbits received either 70 mug/kg estradiol cypionate or vehicle i.m. once a week. Growth plates from the proximal tibia, distal tibia, and distal femur were analyzed after 2, 4, 6, or 8 weeks of treatment. In vehicle-treated animals, there was a gradual senescent decline in tibial growth rate, rate of chondrocyte proliferation, growth plate height, number of proliferative chondrocytes, number of hypertrophic: chondrocytes, size of terminal hypertrophic chondrocytes, and column density. Estrogen treatment accelerated the senescent decline in all of these parameters. In senescent growth plates, epiphyseal fusion was observed to be an abrupt event in which all remaining chondrocytes were rapidly replaced by bone elements. Fusion occurred when the rate of chondrocyte proliferation approached zero. Estrogen caused this proliferative exhaustion and fusion to occur earlier. Our data suggest that (i) epiphyseal fusion is triggered when the proliferative potential of growth plate chondrocytes is exhausted; and (ii) estrogen does not induce growth plate ossification directly; instead, estrogen accelerates the programmed senescence of the growth plate, thus causing earlier proliferative exhaustion and consequently earlier fusion. C1 NICHHD, Dev Endocrinol Branch, NIH, Bethesda, MD 20892 USA. RP Weise, M (reprint author), NICHHD, Dev Endocrinol Branch, NIH, Bethesda, MD 20892 USA. EM wisem@mail.nih.gov NR 41 TC 137 Z9 148 U1 0 U2 9 PU NATL ACAD SCIENCES PI WASHINGTON PA 2101 CONSTITUTION AVE NW, WASHINGTON, DC 20418 USA SN 0027-8424 J9 P NATL ACAD SCI USA JI Proc. Natl. Acad. Sci. U. S. A. PD JUN 5 PY 2001 VL 98 IS 12 BP 6871 EP 6876 DI 10.1073/pnas.121180498 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 440AT UT WOS:000169151500062 PM 11381135 ER PT J AU Egan, MF Goldberg, TE Kolachana, BS Callicott, JH Mazzanti, CM Straub, RE Goldman, D Weinberger, DR AF Egan, MF Goldberg, TE Kolachana, BS Callicott, JH Mazzanti, CM Straub, RE Goldman, D Weinberger, DR TI Effect of COMT Val(108/158) Met genotype on frontal lobe function and risk for schizophrenia SO PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA LA English DT Article ID CATECHOL-O-METHYLTRANSFERASE; DORSOLATERAL PREFRONTAL CORTEX; POSITRON-EMISSION TOMOGRAPHY; WORKING-MEMORY TASK; CEREBRAL BLOOD-FLOW; LINKAGE DISEQUILIBRIUM; PHYSIOLOGICAL DYSFUNCTION; CHROMOSOME 22Q12-Q13.1; SUSCEPTIBILITY LOCUS; DIABETES-MELLITUS AB Abnormalities of prefrontal cortical function are prominent features of schizophrenia and have been associated with genetic risk, suggesting that susceptibility genes for schizophrenia may impact on the molecular mechanisms of prefrontal function. A potential susceptibility mechanism involves regulation of prefrontal dopamine, which modulates the response of prefrontal neurons during working memory. We examined the relationship of a common functional polymorphism (Va(108/158) Met) in the catechol-O-methyltransferase (COMT) gene, which accounts for a 4-fold variation in enzyme activity and dopamine catabolism, with both prefrontally mediated cognition and prefrontal cortical physiology. In 175 patients with schizophrenia, 219 unaffected siblings, and 55 controls, COMT genotype was related in allele dosage fashion to performance on the Wisconsin Card Sorting Test of executive cognition and explained 4% of variance (P = 0.001) in frequency of perseverative errors. Consistent with other evidence that dopamine enhances prefrontal neuronal function, the load of the low-activity Met allele predicted enhanced cognitive performance. We then examined the effect of COMT genotype on prefrontal physiology during a working memory task in three separate subgroups(n = 11-16) assayed with functional MRI, Met allele load consistently predicted a more efficient physiological response in prefrontal cortex. Finally, in a family-based association analysis of 104 trios, we found a significant increase in transmission of the Val allele to the schizophrenic offspring. These data suggest that the COMT Val allele, because it increases prefrontal dopamine catabolism, impairs prefrontal cognition and physiology. and by this mechanism slightly increases risk for schizophrenia. C1 NIMH, Clin Brain Disorders Branch, Bethesda, MD 20892 USA. NIAAA, Neurogenet Lab, Rockville, MD 20852 USA. Virginia Commonwealth Univ, Med Coll Virginia, Dept Psychiat, Richmond, VA 23298 USA. RP Egan, MF (reprint author), NIMH, Clin Brain Disorders Branch, Bldg 10,Ctr Dr, Bethesda, MD 20892 USA. RI Callicott, Joseph/C-9102-2009; Li, Chong/F-4265-2015; Goldman, David/F-9772-2010 OI Callicott, Joseph/0000-0003-1298-3334; Goldman, David/0000-0002-1724-5405 NR 63 TC 1670 Z9 1712 U1 15 U2 90 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 JUN 5 PY 2001 VL 98 IS 12 BP 6917 EP 6922 DI 10.1073/pnas.111134598 PG 6 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 440AT UT WOS:000169151500070 PM 11381111 ER PT J AU Huang, YY Kegeles, LS Bae, SA Hwang, DR Roth, BL Savage, JE Laruelle, M AF Huang, YY Kegeles, LS Bae, SA Hwang, DR Roth, BL Savage, JE Laruelle, M TI Synthesis of potent and selective dopamine D-4 antagonists as candidate radioligands SO BIOORGANIC & MEDICINAL CHEMISTRY LETTERS LA English DT Article ID NONHUMAN PRIMATE BRAIN; RECEPTOR ANTAGONIST; HIGH-AFFINITY; L-745,870; PROFILE AB A series of dopamine D-4 antagonists was synthesized and evaluated as potential candidates for development as positron emission tomography (PET) radioligands. All new compounds display high affinity and selectivity for the D-4 receptors and compounds 5b, 5d, and 5e were identified as candidates for radioligand development. (C) 2001 Elsevier Science Ltd. All rights reserved. C1 Columbia Univ Coll Phys & Surg, Dept Psychiat, New York, NY 10032 USA. Case Western Reserve Univ, Sch Med,NIMH, Psychoact Drug Screening Program, Dept Biochem, Cleveland, OH 44106 USA. Case Western Reserve Univ, Sch Med,NIMH, Psychoact Drug Screening Program, Dept Psychiat, Cleveland, OH 44106 USA. RP Huang, YY (reprint author), Columbia Univ Coll Phys & Surg, Dept Psychiat, 722 W 168th St, New York, NY 10032 USA. RI Roth, Bryan/F-3928-2010 FU NIMH NIH HHS [KO2-MH01366, N01-MH80005, K02-MH01603-01, MH59342-01] NR 20 TC 8 Z9 9 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 JUN 4 PY 2001 VL 11 IS 11 BP 1375 EP 1377 DI 10.1016/S0960-894X(01)00241-4 PG 3 WC Chemistry, Medicinal; Chemistry, Organic SC Pharmacology & Pharmacy; Chemistry GA 438UN UT WOS:000169072800007 PM 11378358 ER PT J AU Taktakishvili, M Neamati, N Pommier, Y Nair, V AF Taktakishvili, M Neamati, N Pommier, Y Nair, V TI Discovery of a nuclease-resistant, non-natural dinucleotide that inhibits HIV-1 integrase SO BIOORGANIC & MEDICINAL CHEMISTRY LETTERS LA English DT Article ID VIRUS TYPE-1 INTEGRASE; RNA AB Integration of HIV viral DNA into human chromosomal DNA catalyzed by HIV integrase is essential for the replication of HIV. Discovery of novel inhibitors of HIV integrase is of considerable significance in approaches to the development of therapeutic agents against AIDS. We have synthesized a new dinucleotide 1 with an internucleotide phosphate bond that is unusually resistant to exonucleases. This compound exhibits potent anti-HIV-1 integrase activity. (C) 2001 Elsevier Science Ltd. All rights reserved. C1 Univ Iowa, Dept Chem, Iowa City, IA 52242 USA. NCI, Mol 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 [AI43181] NR 13 TC 18 Z9 20 U1 0 U2 2 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0960-894X J9 BIOORG MED CHEM LETT JI Bioorg. Med. Chem. Lett. PD JUN 4 PY 2001 VL 11 IS 11 BP 1433 EP 1435 DI 10.1016/S0960-894X(01)00231-1 PG 3 WC Chemistry, Medicinal; Chemistry, Organic SC Pharmacology & Pharmacy; Chemistry GA 438UN UT WOS:000169072800020 PM 11378371 ER PT J AU Chen, WS Norbury, CC Cho, YJ Yewdell, JW Bennink, JR AF Chen, WS Norbury, CC Cho, YJ Yewdell, JW Bennink, JR TI Immunoproteasomes shape immunodominance hierarchies of antiviral CD8(+) T cells at the levels of T cell repertoire and presentation of viral antigens SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Article DE antigen processing; immunodominance; T cells; proteasome; repertoire ID MHC CLASS-I; PROTEASOME SUBUNITS LMP2; DENDRITIC CELLS; LYMPHOCYTE EPITOPE; COMPLEX; MICE; DEGRADATION; SELECTION; MOLECULES; PEPTIDES AB Vertebrates express three cytokine-inducible proteasome subunits that are incorporated in the place of their constitutively synthesized counterparts. There is increasing evidence that the set of peptides generated by proteasomes containing these subunits (immunoproteasomes) differs from that produced by standard proteasomes. In this study, we use mice lacking one of the immunoproteasome subunits (LMP2) to show that immunoproteasomes play an important role in establishing the immunodominance hierarchy of CD8(+) T cells (TCD8+) responding to seven defined determinants in influenza virus. In LMP2(-/-) mice, responses to the two most dominant determinants drop precipitously, whereas responses to two subdominant determinants are greatly enhanced. Adoptive transfer experiments with naive normal and transgenic TCD8+ reveal that the reduced immunogenicity of one determinant (PA(224-233)) can be attributed to decreased generation by antigen presenting cells (APCs), whereas the other determinant (NP366-374) is less immunogenic due to alterations in the TCD8+ repertoire, and not, as reported previously, to the decreased capacity of LMP2(-/-) APCs to generate the determinant. The enhanced response to one of the subdominant determinants (PB1F2(62-70)) correlates with increased generation by LMP2(-/-) virus-infected cells. These findings indicate that in addition to their effects on the presentation of foreign antigens, immunoproteasomes influence TCD8+ responses by modifying the repertoire of responding TCD8+. C1 NIAID, Viral Dis Lab, NIH, Bethesda, MD 20892 USA. RP Yewdell, JW (reprint author), NIAID, Viral Dis Lab, NIH, Rm 211,Bldg 4,4 Ctr Dr,MSC 0440, Bethesda, MD 20892 USA. RI yewdell, jyewdell@nih.gov/A-1702-2012; Chen, Weisan/E-7828-2012 NR 29 TC 145 Z9 150 U1 0 U2 2 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 JUN 4 PY 2001 VL 193 IS 11 BP 1319 EP 1326 DI 10.1084/jem.193.11.1319 PG 8 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 440LV UT WOS:000169177500011 PM 11390439 ER PT J AU Shevach, EM AF Shevach, EM TI Certified professionals: CD4(+)CD25(+) suppressor T cells SO JOURNAL OF EXPERIMENTAL MEDICINE LA English DT Editorial Material ID IMMUNOLOGICAL SELF-TOLERANCE; LYMPHOCYTE-ASSOCIATED ANTIGEN-4; INTESTINAL INFLAMMATION; AUTOIMMUNE-DISEASE; EFFECTOR FUNCTION; DENDRITIC CELLS; IN-VITRO; INDUCTION; INHIBIT; REQUIREMENTS C1 NIAID, Immunol Lab, NIH, Bethesda, MD 20892 USA. RP Shevach, EM (reprint author), NIAID, Immunol Lab, NIH, Bldg 10,Rm 11N315, Bethesda, MD 20892 USA. NR 32 TC 250 Z9 272 U1 0 U2 3 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 JUN 4 PY 2001 VL 193 IS 11 BP F41 EP F45 DI 10.1084/jem.193.11.F41 PG 5 WC Immunology; Medicine, Research & Experimental SC Immunology; Research & Experimental Medicine GA 440LV UT WOS:000169177500001 PM 11390442 ER PT J AU Andrus, MB Turner, TM Updegraff, EP Sauna, ZE Ambudkar, SV AF Andrus, MB Turner, TM Updegraff, EP Sauna, ZE Ambudkar, SV TI Synthesis and analysis of polyethylene glycol linked P-glycoprotein-specific homodimers based on (-)-stipiamide SO TETRAHEDRON LETTERS LA English DT Article DE etherification; polyethylene glycol; dimerization; anticancer compounds; multidrug resistance ID INDEXED COMBINATORIAL LIBRARY; MULTIDRUG-RESISTANCE REVERSAL; NONNATURAL POLYENES; ATP HYDROLYSIS; TRANSPORTER; STIPIAMIDE; BINDING; SITES AB A series of five homodimeric polyethylene glycol (PEG) linked homodimers based on the multidrug resistance reversal agent (-)-stipiamide were made and tested fur their ability to interact with P-glycoprotein, the protein responsible for multidrug resistance, using ATPase and photoaffinity displacement assays. Key reactions include a new alkoxide-mesylate displacement for the assembly of the PEG linkers and a double Sunogashira coupling reaction. (C) 2001 Elsevier Science Ltd. All rights reserved. C1 Brigham Young Univ, Dept Chem & Biochem, Provo, UT 84602 USA. NCI, Cell Biol Lab, Div Basic Sci, NIH, Bethesda, MD 20878 USA. RP Andrus, MB (reprint author), Brigham Young Univ, Dept Chem & Biochem, C100 BNSN, Provo, UT 84602 USA. RI Ambudkar, Suresh/B-5964-2008 NR 27 TC 17 Z9 18 U1 0 U2 4 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0040-4039 J9 TETRAHEDRON LETT JI Tetrahedron Lett. PD JUN 4 PY 2001 VL 42 IS 23 BP 3819 EP 3822 DI 10.1016/S0040-4039(01)00619-0 PG 4 WC Chemistry, Organic SC Chemistry GA 436WP UT WOS:000168959100013 ER PT J AU Gonzalez, FJ Kimura, S AF Gonzalez, FJ Kimura, S TI Understanding the role of xenobiotic-metabolism in chemical carcinogenesis using gene knockout mice SO MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS LA English DT Review DE xenobiotic; cytochrome P450; gene knockout; carcinogenesis ID MICROSOMAL EPOXIDE HYDROLASE; CYP1A2; CAFFEINE; ACETAMINOPHEN; CYP2E1; SUSCEPTIBILITY; EXPRESSION; OXIDATION; SULFOTRANSFERASES; HEPATOTOXICITY AB Most chemical carcinogens require metabolic activation to electrophilic metabolites that are capable of binding to DNA and causing gene mutations. Carcinogen metabolism is carried out by large groups of xenobiotic-metabolizing enzymes that include the phase I cytochromes P450 (P450) and microsomal epoxide hydrolase, and various phase II transferase enzymes. It is extremely important to determine the role P450s play in the carcinogenesis and to establish if they are the rate limiting and critical interface between the chemical and its biological activities. The latter is essential in order to validate the use of rodent models to test safety of chemicals in humans. Since there are marked species differences in expressions and catalytic activities of the multiple P450 forms that activate carcinogens, this validation process becomes especially difficult. To address the role of P450s in whole animal carcinogenesis, mice were produced that lack the P450s known to catalyze carcinogen activation. Mouse lines having disrupted genes encoding the P450s CYP1A2, CYP2E1, and CYP1B1 were developed. Mice lacking expression of microsomal epoxide hydrolase (mEH) and NADPH-quinone oxidoreductase (NQO1) were also made. All of these mice exhibit no gross abnormal phenotypes, suggesting that the xenobiotic-metabolizing enzymes have no critical roles in mammalian development and physiological homeostasis. This explains the occurrence of polymorphisms in xenobiotic-metabolizing enzymes among humans and other mammalian species. However, these null mice do show differences in sensitivities to acute chemical toxicities, thus establishing the importance of xenobiotic metabolism in activation pathways that lead to cell death. Rodent bioassays using null mice and known genotoxic carcinogens should establish whether these enzymes are required for carcinogenesis in an intact animal model. These studies will also provide a framework for the production of transgenic mice and carcinogen bioassay protocols that may be more predictive for identifying the human carcinogens and validate the molecular epidemiological studies ongoing in humans that seek to establish a role for polymorphisms in cancer risk. Published by Elsevier Science B.V. C1 NCI, NIH, Bethesda, MD 20892 USA. RP Gonzalez, FJ (reprint author), NCI, NIH, Bldg 37 Room 3E-24, Bethesda, MD 20892 USA. NR 29 TC 39 Z9 41 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0027-5107 J9 MUTAT RES-FUND MOL M JI Mutat. Res.-Fundam. Mol. Mech. Mutagen. PD JUN 2 PY 2001 VL 477 IS 1-2 SI SI BP 79 EP 87 DI 10.1016/S0027-5107(01)00109-9 PG 9 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology SC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology GA 441WQ UT WOS:000169252900009 PM 11376689 ER PT J AU Deng, CX AF Deng, CX TI Tumorigenesis as a consequence of genetic instability in Brca1 mutant mice SO MUTATION RESEARCH-FUNDAMENTAL AND MOLECULAR MECHANISMS OF MUTAGENESIS LA English DT Article DE Brca1; p53; centrosome; G(2)-M checkpoint; tumorigenesis ID SPLICE VARIANTS BRCA1A; EARLY EMBRYONIC LETHALITY; FAMILIAL BREAST-CANCER; DNA-DAMAGE RESPONSE; INTERACT IN-VIVO; CELL-CYCLE; TUMOR-FORMATION; MESSENGER-RNA; P53 MUTATION; EXPRESSION AB Germline mutations in Brca1 are responsible for most cases of familial breast and ovarian cancers, but somatic mutations in the gene are rarely detected in sporadic rumors. Moreover, mouse embryos carrying Brca1-null mutations or homozygous deletions of Brca1 exon 11 of (Brca1(Delta 11/Delta 11)) die during gestation due to proliferation defects, raising questions about the mechanisms by which Brca1 represses tumor formation. Molecular analysis reveals that these Brca1 mutations cause hypersensitivity to gamma -irradiation and chromosomal abnormalities in embryos and embryonic fibroblast cells (MEFs). Notably, Brca1(Delta 11/Delta 11) MEFs maintain an intact G(1)-S checkpoint, but are defective in G(2)-M checkpoint control. They also contain multiple, functional centrosomes, which lead to unequal chromosome segregation and aneuploidy. These data uncover an essential role for Brca1 in maintaining genetic stability through regulation of centrosome duplication and G(2)-M checkpoint, and provide a molecular basis for its role in tumorigenesis. Finally, we show that conditional mutation of Brca1 in mammary epithelium causes increased apoptosis and abnormal ductal development. Mammary tumor formation in mutant mice occurs after long latency and is associated with p53 mutations. These results are consistent with a model that Brca1 acts as a caretaker gene, whose absence does not directly initiate tumorigenesis, instead, causes genetic instability, which triggers further alterations and ultimately leads to tumor formation. Published by Elsevier Science B.V. C1 NIDDKD, Genet Dev & Dis Branch, NIH, Bethesda, MD 20892 USA. RP Deng, CX (reprint author), NIDDKD, Genet Dev & Dis Branch, NIH, 10-9N105,10 Ctr Dr, Bethesda, MD 20892 USA. RI deng, chuxia/N-6713-2016 NR 58 TC 36 Z9 37 U1 1 U2 1 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 JUN 2 PY 2001 VL 477 IS 1-2 SI SI BP 183 EP 189 DI 10.1016/S0027-5107(01)00119-1 PG 7 WC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology SC Biotechnology & Applied Microbiology; Genetics & Heredity; Toxicology GA 441WQ UT WOS:000169252900019 PM 11376699 ER PT J AU Evdokimov, AG Tropea, JE Routzahn, KM Copeland, TD Waugh, DS AF Evdokimov, AG Tropea, JE Routzahn, KM Copeland, TD Waugh, DS TI Structure of the N-terminal domain of Yersinia pestis YopH at 2.0 angstrom resolution SO ACTA CRYSTALLOGRAPHICA SECTION D-BIOLOGICAL CRYSTALLOGRAPHY LA English DT Article ID PROTEIN-TYROSINE-PHOSPHATASE; MESSENGER-RNA SIGNAL; III SECRETION; FOCAL ADHESIONS; HOST-CELLS; ENTEROCOLITICA; IDENTIFICATION; PROGRAM; CRYSTALLIZATION; REGULATORS AB Yersinia pestis, the causative agent of bubonic plague, injects effector proteins into the cytosol of mammalian cells that enable the bacterium to evade the immune response of the infected organism by interfering with eukaryotic signal transduction pathways. YopH is a modular effector composed of a C-terminal protein tyrosine phosphatase (PTPase) domain and a multifunctional N-terminal domain that not only orchestrates the secretion and translocation of YopH into eukaryotic cells but also binds tyrosine-phosphorylated target proteins to mediate substrate recognition. The crystal structure of the N-terminal domain of YopH (YopH(N); residues 1-130) has been determined at 2.0 Angstrom resolution. The aminoacid sequences that target YopH for secretion from the bacterium and translocation into eukaryotic cells form integral parts of this compactly folded domain. The structure of YopHN bears no resemblance to eukaryotic phosphotyrosine-binding domains, nor is it reminiscent of any known fold. Residues that have been implicated in phosphotyrosine-dependent protein binding are clustered together on one face of YopHN, but the structure does not suggest a mechanism for protein-phosphotyrosine recognition. C1 NCI, Prot Engn Sect, Macromol Crystallog Lab, Ft Detrick, MD 21702 USA. NCI, Struct Biol Core Facil, Ft Detrick, MD 21702 USA. NCI, Prot Chem Core Facil, Ft Detrick, MD 21702 USA. RP Waugh, DS (reprint author), NCI, Prot Engn Sect, Macromol Crystallog Lab, POB B, Ft Detrick, MD 21702 USA. NR 37 TC 39 Z9 39 U1 3 U2 3 PU MUNKSGAARD INT PUBL LTD PI COPENHAGEN PA 35 NORRE SOGADE, PO BOX 2148, DK-1016 COPENHAGEN, DENMARK SN 0907-4449 J9 ACTA CRYSTALLOGR D JI Acta Crystallogr. Sect. D-Biol. Crystallogr. PD JUN PY 2001 VL 57 BP 793 EP 799 DI 10.1107/S0907444901004875 PN 6 PG 7 WC Biochemical Research Methods; Biochemistry & Molecular Biology; Biophysics; Crystallography SC Biochemistry & Molecular Biology; Biophysics; Crystallography GA 446UV UT WOS:000169533300004 PM 11375498 ER PT J AU Saif, MW Bona, R Greenberg, B AF Saif, MW Bona, R Greenberg, B TI AIDS and thrombosis: Retrospective study of 131 HIV-infected patients SO AIDS PATIENT CARE AND STDS LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; PROTEIN-S DEFICIENCY; VENOUS THROMBOSIS; CYTOMEGALO-VIRUS; DISEASE; THROMBOEMBOLISM; ABNORMALITIES; COMPLICATIONS; COAGULATION; INHIBITORS AB The recent literature contains reports of thrombotic episodes occurring in patients with human immunodeficiency virus (HIV) infection and various abnormalities predisposing to a hypercoagulable state have also been reported in such patients. To study the incidence of thrombosis in patients infected with HIV, and to assess the correlation of thrombosis with the degree of immunosuppression as well as the association with active illnesses and neoplasms, we reviewed the charts of 131 patients, which include all the patients with the diagnosis of HIV admitted or seen in the clinic between January 1, 1993, and January 1, 1998. The diagnosis of thrombosis was based on documented reports of venous plethysmography or venography for deep venous thrombosis and ventilation-perfusion scan or pulmonary angiography for pulmonary embolus. Risk factors for thrombotic disease were evaluated including general risk factors such as family history, ambulatory status, medications, and data were also collected regarding CD4 cell counts and the presence of concurrent or remote opportunistic infections, acquired immune deficiency syndrome (AIDS)-related malignancy or other AIDS-related diseases at the time of diagnosis of the thrombotic event. We also reviewed the medical literature via MEDLINE and found 45 cases of patients with HIV who developed thromboembolic complications. We found thrombotic complications in 9 of 37 patients with a CD4 count less than 200 cells/mm(3) and in 1 of the remaining 94 patients with a CD4 count more than 200 cells/mm3. The difference was significant, with p = 0.00004, and the estimated odds of an event given CD4 cell counts less than 200/mm(3) is 29.89 (95% confidence interval). Three patients had abnormalities of anticoagulation proteins. There was a history of opportunistic infections in 5 patients and malignancy in 3 patients. Two patients with autoimmune hemolytic anemia (AIHA) secondary to HIV-infection developed PE upon transfusion of packed red blood cells. The results of this study suggests that AIDS appears to predispose to thrombosis. It also revealed a significant correlation between thrombotic disease and CD4 counts (<200/mm(3)) as well as the presence of opportunistic infections, AIDS-related neoplasms, or autoimmune disorders associated with HIV such as AIHA, Therefore, clinicians caring for these patients should be aware of thromboembolic disease as a possible complication of AIDS. Further studies to elucidate the mechanisms underlying this abnormal hemostatic profile, the epidemiology, and to answer several questions such as should patients with risk factors for HIV infection who develop thromboembolic complications be further evaluated including tests for HIV are warranted. C1 NCI, Med Branch, Natl Naval Med Ctr, NIH, Bethesda, MD 20889 USA. Univ Connecticut, Ctr Hlth, Dept Med, Div Hematol Oncol, Farmington, CT USA. RP Saif, MW (reprint author), NCI, Med Branch, Natl Naval Med Ctr, NIH, Bldg 8,Room 5101,8901 Wisconsin Ave, Bethesda, MD 20889 USA. NR 40 TC 47 Z9 53 U1 0 U2 1 PU MARY ANN LIEBERT INC PUBL PI LARCHMONT PA 2 MADISON AVENUE, LARCHMONT, NY 10538 USA SN 1087-2914 J9 AIDS PATIENT CARE ST JI Aids Patient Care STDS PD JUN PY 2001 VL 15 IS 6 BP 311 EP 320 DI 10.1089/108729101750279687 PG 10 WC Public, Environmental & Occupational Health; Infectious Diseases SC Public, Environmental & Occupational Health; Infectious Diseases GA 446GV UT WOS:000169505600004 PM 11445013 ER PT J AU Patterson, LJ Robey, F Muck, A Van Remoortere, K Aldrich, K Richardson, E Alvord, WG Markham, PD Cranage, M Robert-Guroff, M AF Patterson, LJ Robey, F Muck, A Van Remoortere, K Aldrich, K Richardson, E Alvord, WG Markham, PD Cranage, M Robert-Guroff, M TI A conformational C4 peptide polymer vaccine coupled with live recombinant vector priming is immunogenic but does not protect against rectal SIV challenge SO AIDS RESEARCH AND HUMAN RETROVIRUSES LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; HUMAN MONOCLONAL-ANTIBODY; INFECTED RHESUS-MONKEYS; CD4 BINDING-SITE; ENVELOPE GLYCOPROTEIN; NEUTRALIZING ANTIBODIES; TYPE-1 GP120; IMMUNE-RESPONSE; HIV GP120; MACAQUES AB The conserved, immunogenic CD4 binding site on the viral envelope is an attractive HIV or SIV vaccine candidate. Polymerization of an 18 amino acid segment derived from the C4 domain of SIV gp120 produced a peptide polymer or "peptomer," having an alpha -helical conformation possibly mimicking a proposed structure of the C4 domain in the unbound native protein. The SIV peptomer and native gp120 were compared as subunit boosts following two adenovirus type 5 host range (Ad5hr)-SIVenv recombinant priming immunizations. Both vaccine regimens successfully elicited SIV-specific CTL responses in five of six immunized macaques. Peptomer-boosted macaques exhibited significantly higher envelope-specific T cell proliferative responses than either the gp120-boosted macaques or controls. Peptomer immunization also elicited peptomer and SIV gp120-specific binding antibodies, but only native gp120 boosting elicited SIV neutralizing antibodies. Upon intrarectal challenge with SIVmac32H, all nine macaques became infected. The solely envelope-based vaccine conferred no protection. However, changing the boosting immunogen to the C4 peptomer did not improve protective efficacy in spite of its elicitation of humoral and cellular immune responses, including robust T-helper activity. In spite of the peptomer's strong immunogenicity and potential for induction of broadly protective immune responses, it was not effective as a subunit vaccine. C1 NCI, Basic Res Lab, Bethesda, MD 20892 USA. Natl Inst Dent & Cranofacial Res, Oral & Pharyngeal Canc Branch, Bethesda, MD 20892 USA. NCI, Frederick Canc Res & Dev Ctr, Data Management Serv, Frederick, MD 21702 USA. Adv Biosci Labs Inc, Kensington, MD 20895 USA. Ctr Appl Microbiol & Res, Salisbury SP4 0JG, Wilts, England. RP Robert-Guroff, M (reprint author), NCI, Basic Res Lab, 41 Lib Dr,MSC 5055,Bldg 41,Room D804, Bethesda, MD 20892 USA. EM guroffm@exchange.nih.gov NR 58 TC 18 Z9 20 U1 0 U2 0 PU MARY ANN LIEBERT INC PI NEW ROCHELLE PA 140 HUGUENOT STREET, 3RD FL, NEW ROCHELLE, NY 10801 USA SN 0889-2229 J9 AIDS RES HUM RETROV JI Aids Res. Hum. Retrovir. PD JUN PY 2001 VL 17 IS 9 BP 837 EP 849 DI 10.1089/088922201750252034 PG 13 WC Immunology; Infectious Diseases; Virology SC Immunology; Infectious Diseases; Virology GA 445BE UT WOS:000169437600006 PM 11429125 ER PT J AU Lenfant, C AF Lenfant, C TI Physicians need practical tools to treat the complex problems of overweight and obesity SO AMERICAN FAMILY PHYSICIAN LA English DT Editorial Material ID UNITED-STATES; PREVALENCE C1 NHLBI, NIH, Bethesda, MD 20892 USA. RP Lenfant, C (reprint author), NHLBI, NIH, 31 Ctr Dr,MSC 2480,Bldg 31,Rm 5A52, Bethesda, MD 20892 USA. NR 7 TC 2 Z9 2 U1 0 U2 0 PU AMER ACAD FAMILY PHYSICIANS PI KANSAS CITY PA 8880 WARD PARKWAY, KANSAS CITY, MO 64114-2797 USA SN 0002-838X J9 AM FAM PHYSICIAN JI Am. Fam. Physician PD JUN 1 PY 2001 VL 63 IS 11 BP 2139 EP + PG 2 WC Primary Health Care; Medicine, General & Internal SC General & Internal Medicine GA 442ER UT WOS:000169272700013 PM 11417770 ER PT J AU Ilercil, A Devereux, RB Roman, MJ Paranicas, M O'Grady, MJ Welty, TK Robbins, DC Fabsitz, RR Howard, BV Lee, ET AF Ilercil, A Devereux, RB Roman, MJ Paranicas, M O'Grady, MJ Welty, TK Robbins, DC Fabsitz, RR Howard, BV Lee, ET TI Relationship of impaired glucose tolerance to left ventricular structure and function: The Strong Heart Study SO AMERICAN HEART JOURNAL LA English DT Article ID PRESSURE-OVERLOAD HYPERTROPHY; DEPENDENT DIABETES-MELLITUS; AMERICAN-INDIANS; CARDIOVASCULAR-DISEASE; NORMOTENSIVE CHILDREN; ARTERIAL-HYPERTENSION; RISK-FACTORS; BODY-SIZE; FOLLOW-UP; POPULATION AB Background We have identified increased left ventricular (LV) mass, wall thickness, relative wall thickness, and reduced systolic function in diabetic individuals after adjusting for blood pressure and body mass index. However, the cardiovascular correlates of impaired glucose tolerance (IGT), a precursor of diabetes, are unknown. Methods We compared LV measurements between 457 American Indian participants in the Strong Heart Study with IGT (34% men) by World Health Organization criteria and 888 participants (49% men) with normal glucose tolerance. Results Participants with IGT were older (60 vs 59 years, P < .01), more overweight (body mass index, 32 +/- 6 vs 29 +/- 5 g/m(2)), and had higher systolic blood pressure (129 +/- 20 vs 124 +/- 18 mm Hg, P < .001) and heart rate (67 +/- 10 vs 66 +/- 11 beats/min, P = .011). In univariate analyses, women but not men with IGT had higher LV moss (mean, 150 vs 138 g, P < .001) and cardiac index (2.6 vs 2.5 L/min/m(2), P < .05). LV wall thicknesses and relative wall thickness were greeter in women and men with IGT. Regression analysis, adjusting for multiple covariates in the entire study population, identified independent associations of IGT with higher LV relative wall thicknesses, LV mass/height(2.7), and cardiac output/height(1.83). Conclusions IGT is associated with increased LV wall thickness, mass, and cardiac output independent of effects of relevant covariates. C1 Weill Cornell Med Ctr, New york Presbyterian Hosp, Div Cardiol, New York, NY 10021 USA. Albert Einstein Coll Med, Bronx, NY 10467 USA. Aberdeen Area Tribal Chairmens Hlth Board, Rapid City, SD USA. MedStar Res Inst, Washington, DC USA. NHLBI, Bethesda, MD 20892 USA. Univ Oklahoma, Hlth Sci Ctr, Oklahoma City, OK USA. RP Devereux, RB (reprint author), Weill Cornell Med Ctr, New york Presbyterian Hosp, Div Cardiol, Box 222,525 E 68th St, New York, NY 10021 USA. FU NCRR NIH HHS [M10RR0047]; NHLBI NIH HHS [UO1-HL41654, UO1-HL41642, UO1-HL41652] NR 32 TC 71 Z9 76 U1 1 U2 2 PU MOSBY, INC PI ST LOUIS PA 11830 WESTLINE INDUSTRIAL DR, ST LOUIS, MO 63146-3318 USA SN 0002-8703 J9 AM HEART J JI Am. Heart J. PD JUN PY 2001 VL 141 IS 6 BP 992 EP 998 DI 10.1067/mhj.2001.115302 PG 7 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 438BH UT WOS:000169033500018 PM 11376315 ER PT J AU Bella, JN Devereux, RB Roman, MJ Palmieri, V Liu, JE Paranicas, M Welty, TK Lee, ET Fabsitz, RR Howard, BV AF Bella, JN Devereux, RB Roman, MJ Palmieri, V Liu, JE Paranicas, M Welty, TK Lee, ET Fabsitz, RR Howard, BV CA Strong Heart Study Investigators TI Separate and joint effects of systemic hypertension and diabetes mellitus on left ventricular structure and function in American Indians - (The strong heart study) SO AMERICAN JOURNAL OF CARDIOLOGY LA English DT Article ID PRESSURE-OVERLOAD HYPERTROPHY; CARDIOVASCULAR RISK; BODY-COMPOSITION; MASS; FRAMINGHAM; EPIDEMIOLOGY; CHILDREN; ACCURACY; DISEASE; ADULTS AB Although the association of systemic hypertension (SH) with diabetes mellitus (DM) is well established, the cardiac features and hemodynamic profile of patients with SH and DM diagnosed by American Diabetes Association criteria have not been elucidated. To address this issue, echocardiograms were analyled in 1,025 American Indian participants of the Strong Heart Study with neither DM nor SH, 642 with DM alone, 614 with SH alone, and 874 with SH and DM. In analyses that adjusted for age, gender, body mass index, and heart rate, DM and SH were associated with increased left ventricular (LV) wall thicknesses, with the greatest impact of DM on LV relative wall thickness and of the combination of DM and SH on LV mass (both p <0.001). LV fractional shortening was reduced with SH and SH + DM, midwall shortening was reduced with DM, SH, and their combination, and was reduced in both diabetic groups compared with their nondiabetic counterparts (p <0.001). DM alone was associated with lower measures of LV pump performance (stroke volume, cardiac output, and their indexes) than SH alone. Pulse pressure/stroke index, an indirect measure of arterial stiffness, was elevated in participants with DM or SH alone and most in those with both conditions. There were progressive increases from the reference group to DM alone, SH alone, and DM + SH with regard to prevalences of IV hypertrophy (12% to 19%, 29% and 38%) and subnormal LV myocardial function (7% to 10%, 11% and 18%, both p 0.001). In conclusion, DM and SH each have adverse effects on LV geometry and function, and the combination of SH and DM results in the greatest degree of LV hypertrophy, myocardial dysfunction, and arterial stiffness. (C) 2001 by Excerpta Medica, Inc. C1 Weill Cornell Med Ctr, New York Presbyterian Hosp, Div Cardiol, Dept Med, New York, NY 10021 USA. Aberdeen Area Tribal Chairmens Hlth Board, Rapid City, SD USA. Univ Oklahoma, Sch Publ Hlth Serv, Oklahoma City, OK USA. NHLBI, Div Epidemiol & Clin Applicat, Bethesda, MD 20892 USA. MedStar Res Inst, Washington, DC USA. RP Devereux, RB (reprint author), Weill Cornell Med Ctr, New York Presbyterian Hosp, Div Cardiol, Dept Med, 525 E 68th St, New York, NY 10021 USA. FU NCRR NIH HHS [M10RR0047]; NHLBI NIH HHS [U01-HL41642, U01-HL41654, U01-HL41652] NR 30 TC 84 Z9 90 U1 0 U2 2 PU EXCERPTA MEDICA INC PI NEW YORK PA 650 AVENUE OF THE AMERICAS, NEW YORK, NY 10011 USA SN 0002-9149 J9 AM J CARDIOL JI Am. J. Cardiol. PD JUN 1 PY 2001 VL 87 IS 11 BP 1260 EP 1265 DI 10.1016/S0002-9149(01)01516-8 PG 6 WC Cardiac & Cardiovascular Systems SC Cardiovascular System & Cardiology GA 437YV UT WOS:000169027700006 PM 11377351 ER PT J AU Abylkassimova, Z Kleinerman, R Land, C Weinstock, B Stovall, M Griem, ML AF Abylkassimova, Z Kleinerman, R Land, C Weinstock, B Stovall, M Griem, ML TI Cancer mortality following radiation therapy for peptic ulcer. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20852 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 91 BP S40 EP S40 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400087 ER PT J AU Alavanja, MCR Blomeke, B Vahakangas, KH Welsh, JA Harris, CC Bennett, WP AF Alavanja, MCR Blomeke, B Vahakangas, KH Welsh, JA Harris, CC Bennett, WP TI Glutathione S-transferase (GSTM1), vegetable consumption, residential radon and risk of lung cancer in non-smoking women. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 313 BP S96 EP S96 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400306 ER PT J AU Bai, Y Gao, YT Deng, J Sesterhenn, IA Mostofi, FK Hsing, AW AF Bai, Y Gao, YT Deng, J Sesterhenn, IA Mostofi, FK Hsing, AW TI Family history of cancers and the risk of prostate cancer and benign prostatic hyperplasia in China. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, DCEG, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 731 BP S200 EP S200 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400713 ER PT J AU Balfour, JL Guralnik, JM AF Balfour, JL Guralnik, JM TI Neighborhood socioeconomic deprivation and disability. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIA, EDB Program, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 679 BP S187 EP S187 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400664 ER PT J AU Brenner, A Linet, M Inskip, P AF Brenner, A Linet, M Inskip, P TI History of allergy and risk of brain tumors. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 412 BP S120 EP S120 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400403 ER PT J AU Brown, LM Gridley, G Diehl, SR Winn, DM Harty, LC Bravo-Otero, E Fraumeni, JF Hayes, RB AF Brown, LM Gridley, G Diehl, SR Winn, DM Harty, LC Bravo-Otero, E Fraumeni, JF Hayes, RB TI Family cancer history and susceptibility to oral cancer in Puerto Rico. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 96 BP S41 EP S41 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400093 ER PT J AU Chen, H Miller, B Giovannucci, E Hayes, RB AF Chen, H Miller, B Giovannucci, E Hayes, RB TI Race, socio-economic factors, smoking and drinking and prostate cancer survival. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 748 BP S204 EP S204 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400729 ER PT J AU Chen, H Miller, B Giovannucci, E Hayes, RB AF Chen, H Miller, B Giovannucci, E Hayes, RB TI Anthropometric factors and prostate cancer survival. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 713 BP S196 EP S196 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400697 ER PT J AU Daniels, JL Rowland, AS Longnecker, MP Cook, M Golding, J AF Daniels, JL Rowland, AS Longnecker, MP Cook, M Golding, J TI Dental treatment, fish consumption, and mercury exposure during pregnancy in relation to child neurodevelopment. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIEHS, Res Triangle Pk, NC 27709 USA. NR 0 TC 0 Z9 0 U1 1 U2 3 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 668 BP S184 EP S184 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400654 ER PT J AU David-Beabes, G London, SJ AF David-Beabes, G London, SJ TI Genetic polymorphism of XRCC1 and lung cancer risk among African-Americans and Caucasians. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 Natl Inst Environm Hlth Sci, Res Triangle Pk, NC 27516 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 97 BP S42 EP S42 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400094 ER PT J AU Doody, MM Kampa, D Chimes, K Tarone, RE Sigurdson, AJ Linet, MS AF Doody, MM Kampa, D Chimes, K Tarone, RE Sigurdson, AJ Linet, MS TI Randomized trial of financial incentives for improving response to a mailed questionnaire. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Radiat Epidemiol Branch, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 1 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 260 BP S82 EP S82 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400253 ER PT J AU Freedman, DM Dosemeci, M McGlynn, K AF Freedman, DM Dosemeci, M McGlynn, K TI Sunlight and mortality from breast, overian, colon, prostate, and nonmelanoma skin cancer: A composite death certificate-based case-control study. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Rockville, MD 20852 USA. NR 0 TC 0 Z9 0 U1 0 U2 1 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 722 BP S198 EP S198 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400706 ER PT J AU Grauman, DJ Tarone, RE Jemal, A Devesa, SS AF Grauman, DJ Tarone, RE Jemal, A Devesa, SS TI Interactive cancer mortality mapping and graphing on the World Wide Web. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Rockville, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 114 BP S46 EP S46 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400111 ER PT J AU Gridley, G Winn, DM Brown, LM Weinstein, SJ Diehl, SR Bravo-Otero, E Hayes, RB AF Gridley, G Winn, DM Brown, LM Weinstein, SJ Diehl, SR Bravo-Otero, E Hayes, RB TI The role of dietary factors in the etiology of oral and pharyngeal cancer in Puerto Rico. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 2 U2 2 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 109 BP S45 EP S45 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400105 ER PT J AU Harris, TB Visser, M Goodpaster, B Kritchevsky, S Newman, A Kuller, L Tracy, R Pahor, M Cummings, S Everhart, J AF Harris, TB Visser, M Goodpaster, B Kritchevsky, S Newman, A Kuller, L Tracy, R Pahor, M Cummings, S Everhart, J TI Body fat and circulating cytokines: The health, aging and body composition (Health ABC) study. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIA, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 32 BP S25 EP S25 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400032 ER PT J AU Hoffman, HJ Ko, CW Reed, GW Tambs, K AF Hoffman, HJ Ko, CW Reed, GW Tambs, K TI Aging and other risk factors for noise-induced hearing loss (NIHL): The Nord-Trondelag Health Study, 1995-97. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIDCD, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 11 BP S20 EP S20 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400010 ER PT J AU Huang, WY Winn, DM Bravo-Otero, E Diehl, SR Brown, LM Fraumeni, JF Hayes, RB AF Huang, WY Winn, DM Bravo-Otero, E Diehl, SR Brown, LM Fraumeni, JF Hayes, RB TI Alcohol concentration and oral cancer risk. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 408 BP S119 EP S119 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400399 ER PT J AU Jemal, A Travis, W Tarone, R Travis, L Devesa, S AF Jemal, A Travis, W Tarone, R Travis, L Devesa, S TI The convergence of lung cancer rates between young men and women. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Rockville, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 106 BP S44 EP S44 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400103 ER PT J AU Lacey, JV Mink, PJ Schatzkin, A Sherman, ME Schairer, C AF Lacey, JV Mink, PJ Schatzkin, A Sherman, ME Schairer, C TI Prospective cohort study of ovarian cancer and hormone replacement therapy. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Rockville, MD USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 345 BP S104 EP S104 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400338 ER PT J AU Liu, X Ragan, NB Rogan, WJ AF Liu, X Ragan, NB Rogan, WJ TI Do children with falling blood lead levels have improved cognition? SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIEHS, Res Triangle Pk, NC 27709 USA. RI Rogan, Walter/I-6034-2012 OI Rogan, Walter/0000-0002-9302-0160 NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 669 BP S185 EP S185 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400655 ER PT J AU Liu, X Lai, TJ Guo, YL Guo, NW Rogan, WJ AF Liu, X Lai, TJ Guo, YL Guo, NW Rogan, WJ TI Behavioral problems and cognitive functioning in children with high prenatal PCB exposure: A cohort study. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIEHS, Res Triangle Pk, NC 27709 USA. RI Rogan, Walter/I-6034-2012 OI Rogan, Walter/0000-0002-9302-0160 NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 604 BP S168 EP S168 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400590 ER PT J AU Longnecker, MP Klebanoff, MA Brock, JW Zhou, H Gray, KA Needham, L Wilcox, AJ AF Longnecker, MP Klebanoff, MA Brock, JW Zhou, H Gray, KA Needham, L Wilcox, AJ TI Maternal serum level of DDE and risk of cryptorchidism, hypospadias, and polythelia among male offspring. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIEHS, Res Triangle Pk, NC 27709 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 997 BP S267 EP S267 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400966 ER PT J AU McChesney, R Wilcox, AJ Weinberg, CR Baird, DD AF McChesney, R Wilcox, AJ Weinberg, CR Baird, DD TI Screening with pregnancy tests and detection of conception. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIEHS, Durham, NC 27709 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 561 BP S158 EP S158 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400547 ER PT J AU Miller, BA Hankey, BF Thomas, TL AF Miller, BA Hankey, BF Thomas, TL TI The impact of sociodemographic factors, hormone receptor status, and tumor grade om racial/ethnic differences in tumor stage and size for breast cancer in us women. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20852 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 737 BP S202 EP S202 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400719 ER PT J AU Peters, U Gridley, G Askling, J Linet, M Ekbom, A AF Peters, U Gridley, G Askling, J Linet, M Ekbom, A TI Causes of death in celiac disease patients in a population-based Swedish cohort. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 933 BP S251 EP S251 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400904 ER PT J AU Peters, U Sinha, R Chatterjee, N Subar, A Ziegler, R Bresalier, R Weissfeld, JL Flood, A Hoover, RN Schatzkin, A Hayes, RB AF Peters, U Sinha, R Chatterjee, N Subar, A Ziegler, R Bresalier, R Weissfeld, JL Flood, A Hoover, RN Schatzkin, A Hayes, RB CA PLCO Project Team TI New life for the dietary fiber and colorectal neoplasia hypothesis? Data from the prostate, lung, colorectal, and ovarian cancer (PLCO) trial. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 86 BP S39 EP S39 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400084 ER PT J AU Rooks, R Harris, T AF Rooks, R Harris, T TI Explaining racial differences in cardiovascular disease indicators among older adults: Do socioeconomic resources matter?. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIA, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 180 BP S62 EP S62 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400174 ER PT J AU Rowland, AS Baird, DD Long, S Wegienka, G Harlow, SD Alavanja, M Sandler, DP AF Rowland, AS Baird, DD Long, S Wegienka, G Harlow, SD Alavanja, M Sandler, DP TI Medical conditions and lifestyle factors are associated with menstrual cycle characteristics. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIEHS, Res Triangle Pk, NC 27709 USA. RI 段, 沛沛/C-9902-2010 NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 493 BP S141 EP S141 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400480 ER PT J AU Ruhl, CE Everhart, JE AF Ruhl, CE Everhart, JE TI Leptin concentration and bone mineral density in the United States. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 Social & Sci Syst Inc, Bethesda, MD 20814 USA. NIDDKD, Bethesda, MD 20814 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 40 BP S27 EP S27 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400040 ER PT J AU Salo, PM Xia, J Avol, E Li, Y Johnson, CA Yang, J London, SJ AF Salo, PM Xia, J Avol, E Li, Y Johnson, CA Yang, J London, SJ TI Respiratory symptoms in relation to coal burning and smoking in the home among early adolescents in Wuhan, China. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIEHS, Res Triangle Pk, NC 27516 USA. NR 0 TC 0 Z9 0 U1 1 U2 1 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 466 BP S134 EP S134 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400454 ER PT J AU Schatzkin, A Midthune, D Subar, A Thompson, F Kipnis, V AF Schatzkin, A Midthune, D Subar, A Thompson, F Kipnis, V TI The national institutes of health-American association of retired persons (NIH-AARP) diet and health study: Power to detect diet-cancer associations after adjusting for measurement error. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20892 USA. NR 0 TC 2 Z9 2 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 966 BP S259 EP S259 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400936 ER PT J AU Schroeder, JC Conway, K Taylor, JA AF Schroeder, JC Conway, K Taylor, JA TI P53 mutation, exposure, and genetic susceptibility in a case-control study of bladder cancer. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIEHS, Res Triangle Pk, NC 27709 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 419 BP S122 EP S122 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400408 ER PT J AU Silverman, DT Hoover, RN Brown, LM Swanson, GM Schiffman, M Greenberg, RS Hayes, RB Lillemoe, KD Schoenberg, JB Schwartz, AG Liff, J Pottern, LM Fraumeni, JF AF Silverman, DT Hoover, RN Brown, LM Swanson, GM Schiffman, M Greenberg, RS Hayes, RB Lillemoe, KD Schoenberg, JB Schwartz, AG Liff, J Pottern, LM Fraumeni, JF TI Why do black Americans have a higher risk of pancreatic cancer than white Americans?. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 418 BP S122 EP S122 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400409 ER PT J AU Simonsen, L Morens, D Elixhauser, A Gerber, M Van Raden, M Blackwelder, W AF Simonsen, L Morens, D Elixhauser, A Gerber, M Van Raden, M Blackwelder, W TI Incidence of infant hospitalization for intussusception: Impact of the 1998-99 rotavirus vaccination program in the US SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIAID, NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 617 BP S172 EP S172 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400603 ER PT J AU Stolzenberg-Solomon, R Pietinen, P Taylor, P Virtamo, J Albanes, D AF Stolzenberg-Solomon, R Pietinen, P Taylor, P Virtamo, J Albanes, D TI Medical conditions, anthropometrics, physical activity and pancreatic cancer risk in a cohort of male smokers. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NCI, NIH, Bethesda, MD 20892 USA. RI Albanes, Demetrius/B-9749-2015 NR 0 TC 0 Z9 0 U1 0 U2 1 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 115 BP S46 EP S46 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400110 ER PT J AU Tulloch-Reid, MK Premkumar, A Cochran, CS Reynolds, JC Boston, RC Sumner, AE AF Tulloch-Reid, MK Premkumar, A Cochran, CS Reynolds, JC Boston, RC Sumner, AE TI Sex differences in visceral adipose tissue in African Americans. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 36 BP S26 EP S26 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400036 ER PT J AU Vines, AI McNeilly, MD Stevens, J Hertz-Picciotto, I Bohlig, EM Baird, DD AF Vines, AI McNeilly, MD Stevens, J Hertz-Picciotto, I Bohlig, EM Baird, DD TI Development and reliability of a telephone-administered perceived racism scale (TPRS): A tool for epidemiological use. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIEHS, Res Triangle Pk, NC 27709 USA. NR 0 TC 0 Z9 0 U1 0 U2 2 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 258 BP S82 EP S82 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400252 ER PT J AU Yeo, AET Tanaka, Y Pfeiffer, R Umemura, T Seal, KH Shih, JWK Alter, HJ Edlin, BR O'Brien, TR AF Yeo, AET Tanaka, Y Pfeiffer, R Umemura, T Seal, KH Shih, JWK Alter, HJ Edlin, BR O'Brien, TR TI SEN viruses are common in injection drug users in San Francisco. SO AMERICAN JOURNAL OF EPIDEMIOLOGY LA English DT Meeting Abstract C1 NIH, Dept Transfus Med, Bethesda, MD 20892 USA. RI Pfeiffer, Ruth /F-4748-2011 NR 0 TC 0 Z9 0 U1 0 U2 0 PU OXFORD UNIV PRESS INC PI CARY PA JOURNALS DEPT, 2001 EVANS RD, CARY, NC 27513 USA SN 0002-9262 J9 AM J EPIDEMIOL JI Am. J. Epidemiol. PD JUN 1 PY 2001 VL 153 IS 11 SU S MA 1020 BP S272 EP S272 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 437RB UT WOS:000169006400989 ER PT J AU Egan, CA Smith, EP Taylor, TB Meyer, LJ Samowitz, WS Zone, JJ AF Egan, CA Smith, EP Taylor, TB Meyer, LJ Samowitz, WS Zone, JJ TI Linear IgA bullous dermatosis responsive to a gluten-free diet SO AMERICAN JOURNAL OF GASTROENTEROLOGY LA English DT Article ID DERMATITIS-HERPETIFORMIS; DISEASE; ANTIGEN; ADULTS AB Dermatitis herpetiformis is associated with a gluten-sensitive enteropathy in > 85% of cases. Both the skin lesions and the enteropathy respond to gluten restriction. Linear IgA bullous dermatosis has a much lower prevalence of histological small bowel abnormalities, and lesions are not known to respond to gluten restriction. We report a patient with linear IgA bullous dermatosis and gluten-sensitive enteropathy. This report addresses the issue of whether linear IgA bullous dermatosis can be associated with gluten-sensitive enteropathy. We evaluated the response to gluten restriction and normal diet by following the status of the patient's jejunal biopsies and skin lesions. The patient responded to gluten restriction, as shown by resolution of jejunal abnormalities and skin lesions and subsequently by recurrence of jejunal abnormalities and skin lesions with reinstitution of a gluten-containing diet. This report demonstrates that linear IgA bullous dermatosis can respond to gluten restriction if an underlying gluten-sensitive enteropathy is present. (C) 2001 by Am. Coll. of Gastroenterology. C1 Vet Affairs Med Ctr, Dermatol Sect, Salt Lake City, UT 84148 USA. Vet Affairs Med Ctr, Med Serv, Salt Lake City, UT 84148 USA. Vet Affairs Med Ctr, Ctr Geriatr Res Educ & Clin, Salt Lake City, UT 84148 USA. Univ Utah, Sch Med, Dept Dermatol, Salt Lake City, UT USA. Univ Utah, Sch Med, Dept Pathol, Salt Lake City, UT USA. RP Egan, CA (reprint author), NCI, Dermatol Branch, NIH, Bldg 10,Room 12N238,10 Ctr Dr MSC 1908, Bethesda, MD 20892 USA. FU NIDDK NIH HHS [R01 DK50678-01A1] NR 21 TC 20 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 0002-9270 J9 AM J GASTROENTEROL JI Am. J. Gastroenterol. PD JUN PY 2001 VL 96 IS 6 BP 1927 EP 1929 DI 10.1016/S0002-9270(01)02460-1 PG 3 WC Gastroenterology & Hepatology SC Gastroenterology & Hepatology GA 440XB UT WOS:000169201000043 PM 11419853 ER PT J AU Leroy, JG Seppala, R Huizing, M Dacremont, G De Simpel, H Van Coster, RN Orvisky, E Krasnewich, DM Gahl, WA AF Leroy, JG Seppala, R Huizing, M Dacremont, G De Simpel, H Van Coster, RN Orvisky, E Krasnewich, DM Gahl, WA TI Dominant inheritance of sialuria, an inborn error of feedback inhibition SO AMERICAN JOURNAL OF HUMAN GENETICS LA English DT Article ID N-ACETYLNEURAMINIC ACID; I-CELL DISEASE; SIALIC-ACID; FIBROBLASTS; 2-EPIMERASE; MUTATIONS; GENE AB "French type" sialuria, a presumably dominant disorder that, until now, had been documented in only five patients, manifests with mildly coarse facies, slight motor delay, and urinary excretion of large quantities ( >1 g/d) of free N-acetylneuraminic acid (NeuAc). The basic defect consists of the very rare occurrence of failed feedback inhibition of a rate-limiting enzyme, in this case uridinediphosphate-N-acetylglucosamine (UDP-GlcNAc) 2-epimerase, by a downstream product, in this case cytidine monophosphate (CMP)-NeuAc. We report a new patient with sialuria who has a heterozygous G -->A substitution in nucleotide 848 of the epimerase gene, which results in an R266Q change. The proband's other allele, as expected, had no mutation. However, the heterozygous R266Q mutation was detected in the patient's mother, who has similarly increased urinary levels of free NeuAc, thereby confirming, for the first time, the dominant mode of inheritance of this inborn error. The biochemical diagnosis of the proband was verified by the greatly increased level of free NeuAc in his cultured fibroblasts, the NeuAc distribution, mainly (59%) in the cytoplasm, and by the complete failure of 100 muM CMP-NeuAc to inhibit UDP-GlcNAc 2-epimerase activity in the mutant cells. These findings call for expansion of the phenotype to include adults and for more-extensive assaying of free NeuAc in the urine of children with mild developmental delay. The prevalence of sialuria is probably grossly underestimated. C1 State Univ Ghent, Sch Med, Dept Med Genet, B-9000 Ghent, Belgium. State Univ Ghent, Sch Med, Dept Pediat, B-9000 Ghent, Belgium. NICHHD, Sect Human Biochem Genet, Heritable Disorders Branch, Bethesda, MD 20892 USA. NIMH, Clin Neurosci Branch, Bethesda, MD 20892 USA. NHGRI, Med Genet Branch, NIH, Bethesda, MD 20892 USA. RP Leroy, JG (reprint author), State Univ Ghent, Sch Med, Dept Med Genet, 185 De Pintelaan, B-9000 Ghent, Belgium. NR 21 TC 27 Z9 28 U1 0 U2 2 PU UNIV CHICAGO PRESS PI CHICAGO PA 1427 E 60TH ST, CHICAGO, IL 60637-2954 USA SN 0002-9297 J9 AM J HUM GENET JI Am. J. Hum. Genet. PD JUN PY 2001 VL 68 IS 6 BP 1419 EP 1427 DI 10.1086/320598 PG 9 WC Genetics & Heredity SC Genetics & Heredity GA 439DB UT WOS:000169094600012 PM 11326336 ER PT J AU Alavanja, MCR Sprince, NL Oliver, E Whitten, P Lynch, CF Gillette, PP Logsden-Sacket, N Zwerling, C AF Alavanja, MCR Sprince, NL Oliver, E Whitten, P Lynch, CF Gillette, PP Logsden-Sacket, N Zwerling, C TI Nested case-control analysis of high pesticide exposure events from the Agricultural Health Study SO AMERICAN JOURNAL OF INDUSTRIAL MEDICINE LA English DT Article DE pesticides; pesticide poisoning; accidental exposure; farmers; case-control study AB Background A nested case-control analysis of high pesticide exposure events (HPEEs) was conducted using the Iowa farmers enrolled in the Agricultural Health Study (AHS). Methods In the 12 months of the study, 36 of the 5,970 farmer applicators randomly chosen from the AHS cohort (six per 1,000 farmer applicators per year) met our definition of an HPEE, by reporting "an incident with fertilizers, weed killers, or other pesticides that caused an unusually high personal exposure" resulting in physical symptoms or a visit to a health care provider or hospital. Eligibility criteria were met by 25 HPEE cases and 603 randomly selected controls. Results Significant risk factors for an HPEE included: poor financial condition of the farm which limited the pur-chase of rollover protective structures OR = 4.6 (1.5-16.6), and having a high score on a risk acceptance scale OR = 3.8 (1.4-11.2). Other nonsignificant factors were also identified. Conclusions The limited statistical power of this study necessitates replication of these analyses with a larger sample. Nonetheless, the observed elevated odds ratios of an HPEE provide hypotheses for future studies that may lead to preventive action. Am. J. Ind. Med. 39:557-563, 2001. (C) 2001 Wiley-Liss, Inc. C1 NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Sci Applicat Int Corp, SAIC Frederick, Frederick, MD USA. Univ Iowa, Coll Publ Hlth, Dept Epidemiol, Iowa City, IA USA. Univ Iowa, Coll Publ Hlth, Dept Environm & Occupat Hlth, Iowa City, IA USA. RP Alavanja, MCR (reprint author), NCI, Div Canc Epidemiol & Genet, 6120 Execut Blvd EPS,Rm 8000, Bethesda, MD 20892 USA. NR 17 TC 16 Z9 17 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 0271-3586 J9 AM J IND MED JI Am. J. Ind. Med. PD JUN PY 2001 VL 39 IS 6 BP 557 EP 563 DI 10.1002/ajim.1054 PG 7 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 438DF UT WOS:000169037900003 PM 11385639 ER PT J AU Kirkey, KL Johnson, CC Rybicki, BA Peterson, EL Kortsha, GX Gorell, JM AF Kirkey, KL Johnson, CC Rybicki, BA Peterson, EL Kortsha, GX Gorell, JM TI Occupational categories at risk for Parkinson's disease SO AMERICAN JOURNAL OF INDUSTRIAL MEDICINE LA English DT Article DE occupation; industry; Parkinson's disease; occupational coding; service occupations; agriculture occupations ID EXPOSURE; SMOKING AB Background The etiology of Parkinson's disease (PD) is considered to have a strong environmental component, but relatively few studies have investigated the potential association between occupation and the disease. Methods In a population-based case-control study we collected collected comprehensive occupational histories from all study participants, 144 case and 464 control subjects. Results Chi-square analysis revealed that working in an agricultural occupation increased estimated PD risk (OR = 1.74; 95% CI = 0.85, 3.60). In contrast, a history of ever working in a service occupation was negatively associated with PD risk (OR = 0.69; 95% CI = 0.47, 1.00). Risk estimates were close to one for specific service occupations. Adjusted odds ratios for all non-service occupational and industrial categories were similar, and working in a service occupation was the only significant inverse predictor of PD risk. Conclusions Future investigations focusing on lifestyle factors mid environmental exposures within the agricultural and service occupational categories are warranted. Am. J. Ind. Med. 39:564-571, 2001. (C) 2001 Wiley-Liss, Inc. C1 Henry Ford Hlth Syst, Dept Biostat & Res Epidemiol, Detroit, MI 48202 USA. Henry Ford Hlth Syst, Dept Neurol, Detroit, MI 48202 USA. Wayne State Univ, NIEHS, Ctr Mol & Cellular Toxicol Human Applicat, Detroit, MI USA. RP Kirkey, KL (reprint author), Henry Ford Hlth Syst, Dept Biostat & Res Epidemiol, 1 Ford Pl,3E, Detroit, MI 48202 USA. OI Douwes, Jeroen/0000-0003-3599-4036 NR 20 TC 33 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 0271-3586 J9 AM J IND MED JI Am. J. Ind. Med. PD JUN PY 2001 VL 39 IS 6 BP 564 EP 571 DI 10.1002/ajim.1055 PG 8 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 438DF UT WOS:000169037900004 PM 11385640 ER PT J AU Perretti, M Getting, SJ Solito, E Murphy, PM Gao, JL AF Perretti, M Getting, SJ Solito, E Murphy, PM Gao, JL TI Involvement of the receptor for formylated peptides in the in vivo anti-migratory actions of annexin 1 and its mimetics SO AMERICAN JOURNAL OF PATHOLOGY LA English DT Article ID N-FORMYLPEPTIDE RECEPTOR; NEUTROPHIL; LIPOCORTIN-1; ADHESION; INFLAMMATION; BINDING; INHIBIT; MOUSE; PHENYLALANINE; DEXAMETHASONE AB An innovative avenue for anti-inflammatory therapy is inhibition of neutrophil extravasation by potentiating the action of endogenous anti-inflammatory mediators. The glucocorticoid-inducible protein annexin 1 and derived peptides are effective in inhibiting neutrophil extravasation, Here we tested the hypothesis that an interaction with the receptor for formylated peptide (FPR), so far reported only in vitro, could be the mechanism for this in vivo action. In a model of mouse peritonitis, FPR antagonists abrogated the anti-migratory effects of peptides Ac2-26 and Ac2-12, with a partial reduction in annexin 1 effects. A similar result was obtained in FPR (knock-out) KO mice. Binding of annexin 1 to circulating leukocytes was reduced (>50%) in FPR KO mice. Irt vitro, annexin binding to peritoneal macrophages was also markedly reduced in FPR KO mice, Finally, evidence of direct annexin 1 binding to murine FPR was obtained with HEK-293 cells transfected with the receptor. Overall, these results indicate a functional role for FPR in the anti-migratory effect of annexin 1 and derived peptides. C1 St Bartholomews & Royal London SMD, Div Pharmacol, William Harvey Res Inst, London EC1M 6BQ, England. Univ London Imperial Coll Sci Technol & Med, Sch Med, Dept Neuroendocrinol, London SW7 2AZ, England. NIAID, Host Def Lab, NIH, Bethesda, MD 20892 USA. RP Perretti, M (reprint author), St Bartholomews & Royal London SMD, Div Pharmacol, William Harvey Res Inst, Charterhouse Sq, London EC1M 6BQ, England. NR 34 TC 79 Z9 80 U1 1 U2 6 PU AMER SOC INVESTIGATIVE PATHOLOGY, INC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3993 USA SN 0002-9440 J9 AM J PATHOL JI Am. J. Pathol. PD JUN PY 2001 VL 158 IS 6 BP 1969 EP 1973 DI 10.1016/S0002-9440(10)64667-6 PG 5 WC Pathology SC Pathology GA 439VN UT WOS:000169138400012 PM 11395373 ER PT J AU Kono, H Nakagami, M Rusyn, I Connor, HD Stefanovic, B Brenner, DA Mason, RP Arteel, GE Thurman, RG AF Kono, H Nakagami, M Rusyn, I Connor, HD Stefanovic, B Brenner, DA Mason, RP Arteel, GE Thurman, RG TI Development of an animal model of chronic alcohol-induced pancreatitis in the rat SO AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY LA English DT Article DE medium-chain triglycerides; fibrosis; free radical; enteral feeding ID INTRAGASTRIC ETHANOL INFUSION; INDUCED HEPATIC-FIBROSIS; FREE-RADICAL ADDUCTS; LIVER-DISEASE; DIETARY-FAT; KUPFFER CELLS; DESTRUCTION; INJURY; PATHOGENESIS; NECROSIS AB This study was designed to develop an animal model of alcoholic pancreatitis and to test the hypothesis that the dose of ethanol and the type of dietary fat affect free radical formation and pancreatic pathology. Female Wistar rats were fed liquid diets rich in corn oil (unsaturated fat), with or without a standard or high dose of ethanol, and medium-chain triglycerides (saturated fat) with a high dose of ethanol for 8 wk enterally. The dose of ethanol was increased as tolerance developed, which allowed approximately twice as much alcohol to be delivered in the high-dose group. Serum pancreatic enzymes and histology were normal after 4 wk of diets rich in unsaturated fat, with or without the standard dose of ethanol. In contrast, enzyme levels were elevated significantly by the high ethanol dose. Increases were blunted significantly by dietary saturated fat. Fibrosis and collagen alpha1(I) expression in the pancreas were not detectable after 4 wk of enteral ethanol feeding; however, they were enhanced significantly by the high dose after 8 wk. Furthermore, radical adducts detected by electron spin resonance were minimal with the standard dose; however, the high dose increased carbon-centered radical adducts as well as 4-hydroxynonenal, an index of lipid peroxidation, significantly. Radical adducts were also blunted by similar to 70% by dietary saturated fat. The animal model presented here is the first to demonstrate chronic alcohol-induced pancreatitis in a reproducible manner. The key factors responsible for pathology are the amount of ethanol administered and the type of dietary fat. C1 Univ N Carolina, Dept Pharmacol, Hepatobiol & Toxicol Lab, Chapel Hill, NC 27599 USA. Univ N Carolina, Curriculum Toxicol, Chapel Hill, NC 27599 USA. Univ N Carolina, Dept Med, Chapel Hill, NC 27599 USA. NIEHS, Lab Pharmacol & Chem, Res Triangle Pk, NC 27709 USA. RP Thurman, RG (reprint author), Univ N Carolina, Dept Pharmacol, Hepatobiol & Toxicol Lab, CB 7365,Mary Ellen Jones Bldg, Chapel Hill, NC 27599 USA. RI Rusyn, Ivan/S-2426-2016 NR 34 TC 38 Z9 39 U1 0 U2 1 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0193-1857 J9 AM J PHYSIOL-GASTR L JI Am. J. Physiol.-Gastroint. Liver Physiol. PD JUN PY 2001 VL 280 IS 6 BP G1178 EP G1186 PG 9 WC Gastroenterology & Hepatology; Physiology SC Gastroenterology & Hepatology; Physiology GA 432DJ UT WOS:000168676700017 PM 11352811 ER PT J AU French, S Giulivi, C Balaban, RS AF French, S Giulivi, C Balaban, RS TI Nitric oxide synthase in porcine heart mitochondria: evidence for low physiological activity SO AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY LA English DT Article DE oxidative phosphorylation; oxygen consumption; calcium; ATP; ADP ID OXIDATIVE-PHOSPHORYLATION; METABOLISM; MYOGLOBIN; EXERCISE AB The capacity of isolated porcine heart mitochondria to produce nitric oxide (NO) via mitochondrial NO synthase (NOS) was evaluated. The mitochondrial NOS content and activity (0.2 nmol NO.mg mitochondrial protein(-1).min(-1)) were similar to 10 times lower than previously reported for the rat liver. No evidence for mitochondrial NOS-generated NO was found in mitochondrial suspensions based on the lack of NO production and the lack of effect of either L-arginine or NOS inhibitors on the rate of respiration. The reason that even the low mitochondrial NOS activity did not result in net NO production and metabolic effects is because the mitochondrial metabolic breakdown of NO (1-4 nmol NO.mg mitochondrial protein(-1).min(-1)) was greater than the maximum rate of NO production measured in homogenates. These data suggest that NO production at the mitochondria via NOS is not a significant source of NO in the intact heart and does not regulate cardiac oxidative phosphorylation. C1 NHLBI, Cardiac Energet Lab, NIH, Bethesda, MD 20892 USA. Univ Minnesota, Dept Chem, Duluth, MN 55812 USA. RP French, S (reprint author), NHLBI, Cardiac Energet Lab, NIH, Bldg 10,Rm B1D418, Bethesda, MD 20892 USA. RI Balaban, Robert/A-7459-2009 OI Balaban, Robert/0000-0003-4086-0948 NR 18 TC 63 Z9 65 U1 1 U2 2 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0363-6135 J9 AM J PHYSIOL-HEART C JI Am. J. Physiol.-Heart Circul. Physiol. PD JUN PY 2001 VL 280 IS 6 BP H2863 EP H2867 PG 5 WC Cardiac & Cardiovascular Systems; Physiology; Peripheral Vascular Disease SC Cardiovascular System & Cardiology; Physiology GA 433LV UT WOS:000168761600055 PM 11356646 ER PT J AU Halcox, JPJ Narayanan, S Cramer-Joyce, L Mincemoyer, R Quyyumi, AA AF Halcox, JPJ Narayanan, S Cramer-Joyce, L Mincemoyer, R Quyyumi, AA TI Characterization of endothelium-derived hyperpolarizing factor in the human forearm microcirculation SO AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY LA English DT Article DE vascular tone; endothelium; cytochrome P-450; miconazole; potassium; bradykinin; sodium nitroprusside ID CORONARY ARTERIOLAR DILATION; VASCULAR SMOOTH-MUSCLE; ARACHIDONIC-ACID METABOLITE; CA2+-ACTIVATED K+ CHANNELS; NITRIC-OXIDE; EPOXYEICOSATRIENOIC ACIDS; IN-VIVO; DEPENDENT RELAXATION; BRADYKININ; POTASSIUM AB The identity of endothelium-dependent hyperpolarizing factor (EDHF) in the human circulation remains controversial. We investigated whether EDHF contributes to endothelium-dependent vasomotion in the forearm microvasculature by studying the effect of K+ and miconazole, an inhibitor of cytochrome P-450, on the response to bradykinin in healthy human subjects. Study drugs were infused intra-arterially, and forearm blood flow was measured using strain-gauge plethysmography. Infusion of KCl (0.33 mmol/min) into the brachial artery caused baseline vasodilation and inhibited the vasodilator response to bradykinin, but not to sodium nitroprusside. Thus the incremental vasodilation induced by bradykinin was reduced from 14.3 +/- 2 to 7.1 +/- 2 ml . min(-1) . 100 g(-1) (P< 0.001) after KCl infusion. A similar inhibition of the bradykinin (P = 0.014), but not the sodium nitroprusside (not significant), response was observed with KCl after the study was repeated during preconstriction with phenylephrine to restore resting blood flow to basal values after KCl. Miconazole (0.125 mg/min) did not inhibit endothelium-dependent or -independent responses to ACh and sodium nitroprusside, respectively. However, after inhibition of cyclooxygenase and nitric oxide synthase with aspirin and N-G-monomethyl-L-arginine, the forearm blood flow response to bradykinin (P = 0.003), but not to sodium nitroprusside (not significant), was significantly suppressed by miconazole. Thus nitric oxide- and prostaglandin-independent, bradykinin-mediated forearm vasodilation is suppressed by high intravascular K+ concentrations, indicating a contribution of EDHF. In the human forearm microvasculature, EDHF appears to be a cytochrome P-450 derivative, possibly an epoxyeicosatrienoic acid. C1 NHLBI, Cardiol Branch, NIH, Bethesda, MD 20892 USA. RP Quyyumi, AA (reprint author), NHLBI, Cardiol Branch, NIH, Bldg 10,Rm 7B15,10 Ctr Dr,MSC 1650, Bethesda, MD 20892 USA. NR 46 TC 84 Z9 87 U1 1 U2 1 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0363-6135 J9 AM J PHYSIOL-HEART C JI Am. J. Physiol.-Heart Circul. Physiol. PD JUN PY 2001 VL 280 IS 6 BP H2470 EP H2477 PG 8 WC Cardiac & Cardiovascular Systems; Physiology; Peripheral Vascular Disease SC Cardiovascular System & Cardiology; Physiology GA 433LV UT WOS:000168761600009 PM 11356600 ER PT J AU Kim, HS Kang, SW Rhee, SG Clerch, LB AF Kim, HS Kang, SW Rhee, SG Clerch, LB TI Rat lung peroxiredoxins I and II are differentially regulated during development and by hyperoxia SO AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY LA English DT Article DE antioxidant enzyme; thioredoxin peroxidase; thiol-specific antioxidant; perinatal development ID GENE-EXPRESSION; MAMMALIAN PEROXIREDOXIN; THIOREDOXIN PEROXIDASE; REDUCE PEROXIDES; ISOFORMS; OXYGEN AB Peroxiredoxin I (Prx I) and peroxiredoxin II (Prx II) are found in abundance in the cytoplasm of cells and catalyze the reduction of hydrogen peroxide with the use of electrons provided by thioredoxin. Here we examined Prx I and Prx II expression in rat lung during perinatal development and in response to hyperoxia. Prx I protein increased during late gestation and after birth fell to adult levels; conversely, Prx I mRNA increased after birth. Prx II protein concentration was unchanged in the perinatal period, but Prx II mRNA increased after birth. In response to hyperoxia begun on postnatal day 4, there was no change in Prx II expression; however, Prx I mRNA, protein, and enzymatic activity increased significantly. These data show that 1) Prx I and Prx II are developmentally regulated at the level of translational efficiency and 2) Prx I, but not Prx II, is inducible and is upregulated during the late-gestational preparation for the oxidative stress experienced by the lung at birth and during exposure to hyperoxia in the neonatal period. C1 Georgetown Univ, Med Ctr, Lung Biol Lab, Washington, DC 20007 USA. NHLBI, Lab Cell Signaling, NIH, Bethesda, MD 20892 USA. RP Clerch, LB (reprint author), Georgetown Univ, Med Ctr, Lung Biol Lab, Preclin Sci Bldg,GM12,39000 Reservoir Rd NW, Washington, DC 20007 USA. RI Kim, Han-Suk/D-5788-2011 FU NHLBI NIH HHS [HL-20366, HL-47413] NR 13 TC 17 Z9 20 U1 0 U2 0 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 1040-0605 J9 AM J PHYSIOL-LUNG C JI Am. J. Physiol.-Lung Cell. Mol. Physiol. PD JUN PY 2001 VL 280 IS 6 BP L1212 EP L1217 PG 6 WC Physiology; Respiratory System SC Physiology; Respiratory System GA 431FT UT WOS:000168621700016 PM 11350800 ER PT J AU Petrache, I Verin, AD Crow, MT Birukova, A Liu, F Garcia, JGN AF Petrache, I Verin, AD Crow, MT Birukova, A Liu, F Garcia, JGN TI Differential effect of MLC kinase in TNF-alpha-induced endothelial cell apoptosis and barrier dysfunction SO AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY LA English DT Article DE caspases; cytoskeleton; acute lung injury; permeability; Rho kinase; tumor necrosis factor-alpha ID TUMOR-NECROSIS-FACTOR; LIGHT-CHAIN KINASE; RESPIRATORY-DISTRESS SYNDROME; ACTIN DEPOLYMERIZATION; DNA FRAGMENTATION; GAP FORMATION; PHOSPHORYLATION; DEATH; PERMEABILITY; CLEAVAGE AB Tumor necrosis factor (TNF)-alpha is released in acute inflammatory lung syndromes linked to the extensive vascular dysfunction associated with increased permeability and endothelial cell apoptosis. TNF-alpha induced significant decreases in transcellular electrical resistance across pulmonary endothelial cell monolayers, reflecting vascular barrier dysfunction (beginning at 4 h and persisting for 48 h). TNF-alpha also triggered endothelial cell apoptosis beginning at 4 h, which was attenuated by the caspase inhibitor Z-Val-Ala-Asp-fluoromethylketone. Exploring the involvement of the actomyosin cytoskeleton in these important endothelial cell responses, we determined that TNF-alpha significantly increased myosin light chain (MLC) phosphorylation, with prominent stress fiber and paracellular gap formation, which paralleled the onset of decreases in transcellular electrical resistance and enhanced apoptosis. Reductions in MLC phosphorylation by the inhibition of either MLC kinase (ML-7, cholera toxin) or Rho kinase (Y-27632) dramatically attenuated TNF-alpha -induced stress fiber formation, indexes of apoptosis, and caspase-8 activity but not TNF-alpha -induced barrier dysfunction. These studies indicate a central role for the endothelial cell cytoskeleton in TNF-alpha -mediated apoptosis, whereas TNF-alpha -induced vascular permeability appears to evolve independently of contractile tension generation. C1 Johns Hopkins Univ, Sch Med, Dept Med, Div Pulm & Crit Care Med, Baltimore, MD 21224 USA. NIA, Gerontol Res Ctr, Cardiovasc Sci Lab, Vasc Biol Unit,NIH, Baltimore, MD 21224 USA. RP Garcia, JGN (reprint author), Johns Hopkins Asthma & Allergy Ctr, 5501 Hopkins Bayview Cir,Rm 4B-44, Baltimore, MD 21224 USA. RI Garcia, Joe/E-8862-2010 FU NHLBI NIH HHS [HL-04396, HL-50533, HL-58064] NR 71 TC 100 Z9 108 U1 1 U2 7 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 1040-0605 J9 AM J PHYSIOL-LUNG C JI Am. J. Physiol.-Lung Cell. Mol. Physiol. PD JUN PY 2001 VL 280 IS 6 BP L1168 EP L1178 PG 11 WC Physiology; Respiratory System SC Physiology; Respiratory System GA 431FT UT WOS:000168621700011 PM 11350795 ER PT J AU Wang, WD Kwon, TH Li, CL Flyvbjerg, A Knepper, MA Frokiaer, J Nielsen, S AF Wang, WD Kwon, TH Li, CL Flyvbjerg, A Knepper, MA Frokiaer, J Nielsen, S TI Altered expression of renal aquaporins and Na(+) transporters in rats treated with L-type calcium blocker SO AMERICAN JOURNAL OF PHYSIOLOGY-REGULATORY INTEGRATIVE AND COMPARATIVE PHYSIOLOGY LA English DT Article DE calcium-channel blocker; diuresis; hypertension; natriuresis; urinary concentration mechanism ID THICK ASCENDING LIMB; VASOPRESSIN-ELICITED WATER; MEDULLARY COLLECTING DUCT; PROXIMAL TUBULE; CONCENTRATING DEFECT; CHANNEL EXPRESSION; CONVOLUTED TUBULE; CL COTRANSPORTER; APICAL MEMBRANE; KIDNEY AB Nifedipine, a calcium antagonist, has diuretic and natriuretic properties. However, the molecular mechanisms by which these effects are produced are poorly understood. We examined kidney abundance of aquaporins (AQP1, AQP2, and AQP3) and major sodium transporters [type 3 Na/H exchanger (NHE-3); type 2 Na-Pi cotransporter (NaPi-2); Na-K-ATPase; type 1 bumetanide-sensitive cotransporter (BSC-1); and thiazide-sensitive Na-Cl cotransporter (TSC)] as well as inner medullary abundance of AQP2, phosphorylated-AQP2 (p-AQP2), AQP3, and calcium-sensing receptor (CaR). Rats treated with nifedipine orally (700 mg/kg) for 19 days had a significant increase in urine output, whereas urinary osmolality and solute-free water reabsorption were markedly reduced. Consistent with this, immunoblotting revealed a significant decrease in the abundance of whole kidney AQP2 (47 +/- 7% of control rats, P< 0.05) and in inner medullary AQP2 (60 +/- 7%) as well as in p-AQP2 abundance (17 +/- 6%) in nifedipine-treated rats. In contrast, whole kidney AQP3 abundance was significantly increased (219 +/- 28%). Of potential importance in modulating AQP2 levels, the abundance of CaR in the inner medulla was significantly increased (295 +/- 25%) in nifedipine-treated rats. Nifedipine treatment was also associated with increased urinary sodium excretion. Consistent with this, semiquantitative immunoblotting revealed significant reductions in the abundance of proximal tubule Na 1 transporters: NHE-3 (3 +/- 1%), NaPi-2 (53 +/- 12%), and Na-K-ATPase (74 +/- 5%). In contrast, the abundance of the distal tubule Na-Cl cotransporter (TSC) was markedly increased (240 +/- 29%), whereas BSC-1 in the thick ascending limb was not altered. In conclusion, 1) increased urine output and reduced urinary concentration in nifedipine-treated-rats may, in part, be due to downregulation of AQP2 and p-AQP2 levels; 2) CaR might be involved in the regulation of water reabsorption in the inner medulla collecting duct; 3) reduced expression of proximal tubule Na 1 transporters (NHE-3, NaPi-2, and Na, K-ATPase) may be involved in the increased urinary sodium excretion; and 4) increase in TSC expression may occur as a compensatory mechanism. C1 Aarhus Univ, Inst Anat, Dept Cell Biol, DK-8000 Aarhus C, Denmark. Aarhus Univ Hosp, Inst Expt Clin Res, Dept Clin Physiol, DK-8200 Aarhus, Denmark. Aarhus Univ Hosp, Lab Diabet & Endocrinol M, DK-8000 Aarhus C, Denmark. Dongguk Univ, Sch Med, Dept Physiol, Kyungju 780714, South Korea. NHLBI, Kidney & Electrolyte Metab Lab, NIH, Bethesda, MD 20892 USA. RP Nielsen, S (reprint author), Aarhus Univ, Inst Anat, Dept Cell Biol, DK-8000 Aarhus C, Denmark. EM sn@ana.au.dk FU Intramural NIH HHS [Z01 HL001285-21, Z99 HL999999] NR 51 TC 7 Z9 8 U1 1 U2 5 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 0363-6119 J9 AM J PHYSIOL-REG I JI Am. J. Physiol.-Regul. Integr. Comp. Physiol. PD JUN PY 2001 VL 280 IS 6 BP R1632 EP R1641 PG 10 WC Physiology SC Physiology GA 432DK UT WOS:000168676800006 PM 11353665 ER PT J AU Hager, H Kwon, TH Vinnikova, AK Masilamani, S Brooks, HL Frokiaer, J Knepper, MA Nielsen, S AF Hager, H Kwon, TH Vinnikova, AK Masilamani, S Brooks, HL Frokiaer, J Knepper, MA Nielsen, S TI Immunocytochemical and immunoelectron microscopic localization of alpha-, beta-, and gamma-ENaC in rat kidney SO AMERICAN JOURNAL OF PHYSIOLOGY-RENAL PHYSIOLOGY LA English DT Article DE aldosterone; collecting duct; urothelium; epithelial sodium channel; intracellular trafficking; sodium transport ID EPITHELIAL SODIUM-CHANNEL; COLLECTING DUCT; WATER CHANNEL; APICAL MEMBRANE; MEDIATED REGULATION; ALDOSTERONE; EXPRESSION; VASOPRESSIN; SUBUNIT; NA AB Epithelial sodium channel (ENaC) subunit (alpha, beta, and gamma) mRNA and protein have been localized to the principal cells of the connecting tubule (CNT), cortical collecting duct (CCD), and outer medullary collecting duct (OMCD) in rat kidney. However, the subcellular localization of ENaC subunits in the principal cells of these cells is undefined. The cellular and subcellular localization of ENaC subunits in rat kidney was therefore examined. Immunocytochemistry demonstrated the presence of all three subunits in principal cells of the CNT, CCD, OMCD, and IMCD. In cortex and outer medulla, confocal microscopy demonstrated a difference in the subcellular localization of subunits. alpha -ENaC was localized mainly in a zone in the apical domains, whereas beta- and gamma -ENaC were found throughout the cytoplasm. Immunoelectron microscopy confirmed the presence of ENaC subunits in both the apical plasma membrane and intracellular vesicles. In contrast to the labeling pattern seen in cortex, alpha -ENaC labeling in IMCD cells was distributed throughout the cytoplasm. In the urothelium covering pelvis, ureters, and bladder, immunoperoxidase and confocal microscopy revealed differences the presence of all ENaC subunits. As seen in CCD, alpha -ENaC was present in a narrow zone near the apical plasma membrane, whereas band gamma -ENaC were dispersed throughout the cytoplasm. In conclusion, all three subunits of ENaC are expressed throughout the collecting duct (CD), including the IMCD as well as in the urothelium. The intracellular vesicular pool in CD principal cells suggests ENaC trafficking as a potential mechanism for the regulation of Na(+) reabsorption. C1 Aarhus Univ, Inst Anat, Dept Cell Biol, DK-8000 Aarhus C, Denmark. Dongguk Univ, Sch Med, Dept Physiol, Kyungju 780714, South Korea. McGuire Vet Affairs Med Ctr, Res Serv, Richmond, VA 23249 USA. NHLBI, Kidney & Electrolyte Metab Lab, NIH, Bethesda, MD 20892 USA. Aarhus Univ Hosp, Dept Clin Physiol, DK-8200 Aarhus N, Denmark. Inst Expt Clin Res, DK-8200 Aarhus N, Denmark. RP Nielsen, S (reprint author), Aarhus Univ, Inst Anat, Dept Cell Biol, DK-8000 Aarhus C, Denmark. EM sn@ana.au.dk FU Intramural NIH HHS [Z01 HL001285-21, Z99 HL999999] NR 38 TC 115 Z9 117 U1 0 U2 2 PU AMER PHYSIOLOGICAL SOC PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814 USA SN 1931-857X J9 AM J PHYSIOL-RENAL JI Am. J. Physiol.-Renal Physiol. PD JUN PY 2001 VL 280 IS 6 BP F1093 EP F1106 PG 14 WC Physiology; Urology & Nephrology SC Physiology; Urology & Nephrology GA 432DL UT WOS:000168676900016 PM 11352848 ER PT J AU Fee, E Brown, TM AF Fee, E Brown, TM TI Walter J. Lear - Voices from the past SO AMERICAN JOURNAL OF PUBLIC HEALTH LA English DT Biographical-Item C1 Natl Lib Med, Hist Med Div, Bethesda, MD 20894 USA. RP Fee, E (reprint author), Natl Lib Med, Hist Med Div, 8600 Rockville Pike, Bethesda, MD 20894 USA. NR 0 TC 1 Z9 1 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 JUN PY 2001 VL 91 IS 6 BP 902 EP 902 PG 1 WC Public, Environmental & Occupational Health SC Public, Environmental & Occupational Health GA 461BQ UT WOS:000170345400017 ER PT J AU O'Grady, NP Preas, HL Pugin, J Fiuza, C Tropea, M Reda, D Banks, SM Suffredini, AF AF O'Grady, NP Preas, HL Pugin, J Fiuza, C Tropea, M Reda, D Banks, SM Suffredini, AF TI Local inflammatory responses following bronchial endotoxin instillation in humans SO AMERICAN JOURNAL OF RESPIRATORY AND CRITICAL CARE MEDICINE LA English DT Article ID TUMOR-NECROSIS-FACTOR; RESPIRATORY-DISTRESS-SYNDROME; BRONCHOALVEOLAR LAVAGE FLUID; COLONY-STIMULATING FACTOR; ALVEOLAR MACROPHAGES; INNATE IMMUNITY; IN-VIVO; ANTIINFLAMMATORY CYTOKINE; INTERLEUKIN-6 IL-6; LUNG INJURY AB To study local lung inflammation, 34 subjects had endotoxin (1-4 ng/kg) instilled into a lung segment and saline instilled into a contralateral segment followed by bronchoalveolar lavage (BAL) at 2 h, 6 h, 24 h, or 48 h. Endotoxin instillation resulted in a focal inflammatory response with a distinct time course. An early phase (2 h to 6 h) revealed an increase in neutrophils (p = 0.0001) with elevated cytokines (tumor necrosis factor [TNF]-alpha, TNF receptors [TNFR], interleukin [IL]-1 beta, IL-1 receptor antagonist, IL-6, granulocyte-colony-stimulating factor [G-CSF], all p less than or equal to 0.002, but no change in IL-10) and chemokines (IL-8, epithelial neutrophil activating protein-78, monocyte chemotactic protein-1, macrophage inflammatory protein [MIP]-1 alpha, MIP-1 beta, all p less than or equal to 0.001, but no change in growth-regulated peptide-alpha). A later phase (24 h to 48 h) showed increased neutrophils, macrophages, monocytes, and lymphocytes (all p less than or equal to 0.02), and a return to basal levels of most mediators. Elevated levels of inflammatory markers (TNFR1, TNFR2, L-selectin, lactoferrin, and myeloperoxidase) persisted in the BAL at 48 h (p less than or equal to 0.001). Increased permeability to albumin occurred throughout both phases (p = 0.001). Blood C-reactive protein, serum amyloid A, IL-6, IL-1ra, G-CSF, but not TNF-alpha increased by 8 h (all p less than or equal to 0.008). The local pulmonary inflammatory response to endotoxin has a unique qualitative and temporal profile of inflammation compared with previous reports of intravenous endotoxin challenges. This model provides a means to investigate factors that initiate, amplify, and resolve local lung inflammation. C1 NIH, Dept Crit Care Med, Warren G Magnuson Clin Ctr, CC, Bethesda, MD 20892 USA. Univ Hosp Geneva, Div Med Crit Care, Geneva, Switzerland. RP Suffredini, AF (reprint author), NIH, Dept Crit Care Med, Warren G Magnuson Clin Ctr, CC, Bldg 10,Room 7D-43,10 Ctr Dr, Bethesda, MD 20892 USA. NR 45 TC 144 Z9 146 U1 0 U2 0 PU AMER THORACIC SOC PI NEW YORK PA 1740 BROADWAY, NEW YORK, NY 10019-4374 USA SN 1073-449X J9 AM J RESP CRIT CARE JI Am. J. Respir. Crit. Care Med. PD JUN PY 2001 VL 163 IS 7 BP 1591 EP 1598 PG 8 WC Critical Care Medicine; Respiratory System SC General & Internal Medicine; Respiratory System GA 443JE UT WOS:000169337400019 PM 11401879 ER PT J AU Metheny-Barlow, LJ Flynn, B van Gijssel, HE Marrogi, A Gerwin, BI AF Metheny-Barlow, LJ Flynn, B van Gijssel, HE Marrogi, A Gerwin, BI TI Paradoxical effects of platelet-derived growth factor-A overexpression in malignant mesothelioma - Antiproliferative effects in vitro and tumorigenic stimulation in vivo SO AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY LA English DT Article ID FACTOR INDUCES APOPTOSIS; PDGF ALPHA-RECEPTORS; SMOOTH-MUSCLE CELLS; PLEURAL MESOTHELIOMA; SIGNAL-TRANSDUCTION; IN-VITRO; EXPRESSION; ASBESTOS; LINES; BETA AB Malignant mesothelioma is associated with asbestos exposure and remains resistant to all therapeutic intervention. Previous studies have suggested an enhancing role for platelet-derived growth factor (PDGF) in mesothelial tumorigenicity, although the mechanism by which PDGF facilitates tumorigenicity is unknown. Here, we evaluate the contribution of PDGF-A expression to mesothelial tumorigenicity using ectopic modulation of PDGF-A expression. We find, in accordance with other reports, that the receptor for PDGF-A, although expressed at high levels in normal human mesothelial cells, is not easily detectable in mesothelioma. Further, we show that PDGF-A overexpression is responsible for autocrine downregulation of its receptor. Our data indicate, surprisingly, that for mesothelioma cells in vitro, high-level activation of a PDGF-A-PDGF receptor loop is antiproliferative whereas abrogation of PDGF-A expression stimulates growth. These data suggest that PDGF-A does not contribute to tumorigenicity by autocrine stimulation of proliferation. In contrast, increased PDGF-A expression in vivo increases tumor incidence and growth rate and decreases the latency period to tumor formation whereas abrogation of PDGF-A expression decreases tumor incidence and increases latency. Thus, the tumorigenic effect of PDGF-A must act through paracrine mechanisms relevant at early stages of tumor initiation. C1 NCI, Human Carcinogenesis Lab, Bethesda, MD 20892 USA. NCI, Cellular Carcinogenesis & Tumor Promot Lab, Div Basic Sci, Bethesda, MD 20892 USA. RP Metheny-Barlow, LJ (reprint author), Georgetown Univ, Lombardi canc Ctr, Res Bldg E301,3970 Reservoir Rd NW, Washington, DC 20007 USA. RI Metheny-Barlow, Linda/C-3123-2011 NR 53 TC 19 Z9 19 U1 0 U2 0 PU AMER THORACIC SOC PI NEW YORK PA 1740 BROADWAY, NEW YORK, NY 10019-4374 USA SN 1044-1549 J9 AM J RESP CELL MOL JI Am. J. Respir. Cell Mol. Biol. PD JUN PY 2001 VL 24 IS 6 BP 694 EP 702 PG 9 WC Biochemistry & Molecular Biology; Cell Biology; Respiratory System SC Biochemistry & Molecular Biology; Cell Biology; Respiratory System GA 447XR UT WOS:000169598200008 PM 11415934 ER PT J AU Liu, F Verin, AD Wang, P Day, R Wersto, RP Chrest, FJ English, DK Garcia, JGN AF Liu, F Verin, AD Wang, P Day, R Wersto, RP Chrest, FJ English, DK Garcia, JGN TI Differential regulation of sphingosine-1-phosphate and VEGF-induced endothelial cell chemotaxis - Involvement of G(i alpha 2)-linked rho kinase activity SO AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECULAR BIOLOGY LA English DT Article ID PROTEIN-COUPLED RECEPTORS; FOCAL ADHESION KINASE; MYOSIN LIGHT-CHAIN; SPHINGOSINE 1-PHOSPHATE; LYSOPHOSPHATIDIC ACID; GROWTH-FACTOR; ACTIN REORGANIZATION; SIGNAL-TRANSDUCTION; PHARMACOLOGICAL PROPERTIES; MEDIATED PHOSPHORYLATION AB We compared stimulus-coupling pathways involved in bovine pulmonary artery (PA) and lung microvascular endothelial cell migration evoked by sphingosine-l-phosphate (S1P), a potent bioactive lipid released from activated platelets, and by vascular endothelial growth factor (VEGF), a well-recognized angiogenic factor. S1P-induced endothelial cell migration was maximum at 1 muM (similar to 8-fold increase with PA endothelium) and surpassed the maximal response evoked by either VEGF (10 ng/ml) (similar to 2.5-fold increase) or hepatocyte growth factor (HGF) (similar to 2.5-foId increase). Migration induced by S1P, but not by VEGF, was significantly inhibited by treatment with antisense oligonucleotides directed to Edg-1 and Edg-3 (endothelial differentiation gene) S1P receptors and by G protein modification. These strategies included pretreatment with pertussis toxin, or transfection with mini-genes encoding a py subunit inhibitory peptide of the beta -adrenergic receptor kinase, or an 11-amino-acid peptide that inhibits G(1 alpha2) signaling. Various strategies to interrupt Rho family signaling, including C-3 exotoxin, dominant/negative Rho, or the addition of Y27632, a cell-permeable Rho kinase inhibitor, significantly attenuated S1P- but not VEGF-induced migration. Conversely, pharmacologic Inhibition of either myosin light chain kinase, src family tyrosine kinases, or phosphatidylinositol-3' kinase reduced basal endothelial cell migration and abolished VEGF-induced endothelial cell migration but did not inhibit the increase in S1P-induced migration. Whereas VEGF and S1P increased both p42/p44 extracellular regulated kinase and p38 mitogen-activated protein (MAP) kinase activities, only p38 MAP kinase inhibition significantly reduced VEGF- and S1P-stimulated migration. These data confirm S1P as a potent endothelial cell chemoattractant through G(1 alpha2)-coupled Edg receptors linked to Rho-associated kinase and p38 MAP kinase activation. The divergence in signaling pathways evoked by S1P and VEGF suggests complex and agonist-specific regulation of endothelial cell angiogenic responses. C1 Johns Hopkins Univ, Sch Med, Div Pulm & Crit Care Med, Baltimore, MD 21205 USA. NIA, Baltimore, MD 21224 USA. Methodist Res Inst, Indianapolis, IN USA. Tufts Univ, Boston, MA 02111 USA. RP Garcia, JGN (reprint author), Johns Hopkins Asthma & Allergy Ctr, 5501 Hopkins Bayview Circle,4B-77, Baltimore, MD 21224 USA. RI Garcia, Joe/E-8862-2010 FU NHLBI NIH HHS [HL 50333, HL58064] NR 51 TC 126 Z9 126 U1 0 U2 2 PU AMER THORACIC SOC PI NEW YORK PA 1740 BROADWAY, NEW YORK, NY 10019-4374 USA SN 1044-1549 J9 AM J RESP CELL MOL JI Am. J. Respir. Cell Mol. Biol. PD JUN PY 2001 VL 24 IS 6 BP 711 EP 719 PG 9 WC Biochemistry & Molecular Biology; Cell Biology; Respiratory System SC Biochemistry & Molecular Biology; Cell Biology; Respiratory System GA 447XR UT WOS:000169598200010 PM 11415936 ER PT J AU Mirsky, AF AF Mirsky, AF TI Herbert Charles Lansdell (1922-2000). Obituary SO AMERICAN PSYCHOLOGIST LA English DT Biographical-Item C1 NIMH, Bethesda, MD 20892 USA. RP Mirsky, AF (reprint author), NIMH, Bethesda, MD 20892 USA. NR 1 TC 0 Z9 0 U1 0 U2 0 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 0003-066X J9 AM PSYCHOL JI Am. Psychol. PD JUN-JUL PY 2001 VL 56 IS 6-7 BP 518 EP 518 PG 1 WC Psychology, Multidisciplinary SC Psychology GA 463CL UT WOS:000170459200009 ER PT J AU Menter, DG Hoque, A Motiwala, N Sahin, AA Sneige, N Lieberman, R Lippman, SM AF Menter, DG Hoque, A Motiwala, N Sahin, AA Sneige, N Lieberman, R Lippman, SM TI Computerized image analysis of Ki-67 in ductal breast carcinoma in situ SO ANALYTICAL AND QUANTITATIVE CYTOLOGY AND HISTOLOGY LA English DT Article DE Ki-67 antigen; ductal breast carcinoma in situ; image analysis, computer-assisted ID PROLIFERATION-RELATED ANTIGEN; MONOCLONAL-ANTIBODY KI-67; CELL-PROLIFERATION; IN-SITU; GROWTH FRACTION; CANCER CHEMOPREVENTION; BIOLOGICAL MARKERS; NUCLEAR ANTIGEN; CYCLE; LOCALIZATION AB OBJECTIVE: To develop and determine the staining protocols and computerized image analysis methods that are the most effective combination for performing quantitative analysis of Ki-67. STUDY DESIGN: We compared conventional bright-field light microscopy and refractive optical enhancement methods in combination with various immunodetection and filter enhancement methods, including immunogold in combination with epipolarization refractive optics and enzymatic conversion of chromogenic substrates in combination with optical filter enhancement. Initial Ki-67 tests were performed on lymph node tissues and cultured human breast cells and then applied to 200 ductal carcinoma in situ (DCIS) samples. DCIS acini were digitally acquried, and a region of interest was manually outlined in each one with a digital stylus to include only the cellular component; then the Ki-67 staining index was quantified by segmentation analysis. RESULTS: Although combining epipolarization analysis with immunohistogold staining was the most sensitive detection method, nonspecific binding was too high. The streptavidin-horseradish-peroxide enzymatic conversion of 3,3 '-diaminobenzidine (DAB) in combination with optical enhancement filters was the most effective method tested. Ki-67 stain was associated with dense fibrillar structures of the nucleoli in the less intensely staining nuclei and was most intense in paired nuclei. CONCLUSION: The method of measuring Ki-67 expression by DAB staining combined with optical enhancement filters and quantification via computer assisted image analysis techniques produced objective and reproducible results. As such, this method can offer (1) less intraobserver and interobserver variability, (2) a digital archival record, and (3) a baseline for digital exchange of information between studies. C1 Univ Texas, MD Anderson Canc Ctr, Dept Clin Canc Prevent, Houston, TX 77030 USA. Univ Texas, MD Anderson Canc Ctr, Dept Pathol, Houston, TX 77030 USA. NCI, Div Canc Prevent, Prostate & Urol Canc Res Grp, Rockville, MD USA. RP Lippman, SM (reprint author), Univ Texas, MD Anderson Canc Ctr, Dept Clin Canc Prevent, 1515 Holcombe Blvd,Box 236, Houston, TX 77030 USA. FU NCI NIH HHS [CN-65004, CA57730] NR 41 TC 6 Z9 6 U1 0 U2 0 PU SCI PRINTERS & PUBL INC PI ST LOUIS PA PO DRAWER 12425 8342 OLIVE BLVD, ST LOUIS, MO 63132 USA SN 0884-6812 J9 ANAL QUANT CYTOL JI Anal. Quant. Cytol. Histol. PD JUN PY 2001 VL 23 IS 3 BP 218 EP 228 PG 11 WC Cell Biology SC Cell Biology GA 446KV UT WOS:000169513200008 PM 11444192 ER PT J AU McPhie, P AF McPhie, P TI Circular dichroism studies on proteins in films and in solution: Estimation of secondary structure by g-factor analysis SO ANALYTICAL BIOCHEMISTRY LA English DT Article DE circular dichroism; g-factor; protein secondary structure ID CONFORMATION; SPECTRA AB Estimates of the secondary structure of a protein in solution are made by mathematical analyses of its circular dichroism (CD) spectrum below 240 nm. All current procedures require accurate determination of the concentration of the protein sample. Insoluble proteins, such as prions or amyloid, are examined as thin films or gels, but concentrations cannot be precisely defined. The ratio of a sample's CD and absorbance signals is the g-factor, an intensive property. The g-factor spectra of 19 soluble, unconjugated proteins of known structures were measured and used to derive basis spectra, characteristic of the four major structural elements, helix, sheet, turn, and remainder. Using these, the g-factor spectra of other unconjugated proteins, measured in solution or as films, can be analyzed by linear regression to give good estimates of their secondary structures when protein concentration cannot be determined. (C) 2001 Academic Press. C1 NIDDK, Lab Biochem & Genet, NIH, Bethesda, MD 20892 USA. RP McPhie, P (reprint author), NIDDK, Lab Biochem & Genet, NIH, Bldg 8,Room 215, Bethesda, MD 20892 USA. NR 27 TC 34 Z9 36 U1 6 U2 17 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0003-2697 J9 ANAL BIOCHEM JI Anal. Biochem. PD JUN 1 PY 2001 VL 293 IS 1 BP 109 EP 119 DI 10.1006/abio.2001.5113 PG 11 WC Biochemical Research Methods; Biochemistry & Molecular Biology; Chemistry, Analytical SC Biochemistry & Molecular Biology; Chemistry GA 441PM UT WOS:000169238800016 PM 11373086 ER PT J AU Corle, DK Sharbaugh, C Mateski, DJ Coyne, T Paskett, ED Cahill, J Daston, C Lanza, E Schatzkin, A AF Corle, DK Sharbaugh, C Mateski, DJ Coyne, T Paskett, ED Cahill, J Daston, C Lanza, E Schatzkin, A CA PPT Study Grp TI Self-rated quality of life measures: Effect of change to a low-fat, high-fiber, fruit and vegetable enriched diet SO ANNALS OF BEHAVIORAL MEDICINE LA English DT Article ID CORONARY HEART-DISEASE; PREVENTION TRIAL; OF-LIFE; CHOLESTEROL CONCENTRATIONS; SERUM-CHOLESTEROL; RISK-FACTORS; INTERVENTION; HEALTH; DEATH; REHABILITATION AB The Polyp Prevention Trial (PPT) was a multicenter randomized clinical trial to determine the effect of a low-fat (20% of energy from fat), high-fiber (18 g/1000 kcal/day), high-fruit/vegetable (3.5 servings/1000 kcal/day) eating plan on the recurrence of large bowel adenomatous polyps. The PPT provided an opportunity to examine the impact of dietary changes on quality of life. At baseline and annually for 4 years, participants in the Quality of Life Substudy of PPT completed a Quality of Life Factors (QF) Questionnaire, a modified Block-National Cancer Institute Food Frequency Questionnaire, and a Health and Lifestyle Questionnaire. The 51-item QF Questionnaire assessed changes in nine domains: taste, convenience, cost, self-care, social, health assessment, health belief health action, and life satisfaction. The analysis compared annual changes in domain scores for intervention (n = 194) and control (n = 200) participants. At Year 1,363 (92%) completed a questionnaire, and 325 (82%) participants completed a Year 4 questionnaire. There were no statistically significant differences between treatment groups in the change from baseline to Year I for the convenience, cost, taste, health assessment, and life satisfaction domains. At Year 1, intervention participants rated the self-care (p < .001), health belief (p = .021), and health action (p < .001) domains significantly higher and the social domain significantly lower (p < .001) than control participants, These changes were consistent through Years 2, 3, and 4. This study provides evidence that, given appropriate support, five-living individuals can successfully alter their eatingpatterns in multiple ways without a negative impact on quality of life. C1 NCI, Bethesda, MD 20892 USA. Univ Pittsburgh, Pittsburgh, PA 15260 USA. Wake Forest Univ, Winston Salem, NC 27109 USA. RP Corle, DK (reprint author), NCI, Execut Plaza N,Suite 3131,6130 Execut Blvd,MSC 73, Bethesda, MD 20892 USA. FU NCI NIH HHS [2SC05221] NR 41 TC 13 Z9 13 U1 1 U2 6 PU LAWRENCE ERLBAUM ASSOC INC PI MAHWAH PA 10 INDUSTRIAL AVE, MAHWAH, NJ 07430-2262 USA SN 0883-6612 J9 ANN BEHAV MED JI Ann. Behav. Med. PD SUM PY 2001 VL 23 IS 3 BP 198 EP 207 DI 10.1207/S15324796ABM2303_7 PG 10 WC Psychology, Multidisciplinary SC Psychology GA 457DE UT WOS:000170121200007 PM 11495220 ER PT J AU Wagner, KR Hamed, S Hadley, DW Gropman, AL Burstein, AH Escolar, DM Hoffman, EP Fischbeck, KH AF Wagner, KR Hamed, S Hadley, DW Gropman, AL Burstein, AH Escolar, DM Hoffman, EP Fischbeck, KH TI Gentamicin treatment of Duchenne and Becker muscular dystrophy due to nonsense mutations SO ANNALS OF NEUROLOGY LA English DT Article ID AMINOGLYCOSIDE ANTIBIOTICS; HUMAN-CELLS; GLYCOPROTEIN COMPLEX; ESCHERICHIA-COLI; STOP CODON; MDX MICE; DMD GENE; SUPPRESSION; UAG; PHARMACOKINETICS AB Aminoglycosides have previously been shown to suppress nonsense mutations, allowing translation of full-length proteins in vitro and in animal models. In the mdx mouse, where muscular dystrophy is due to a nonsense mutation in the dystrophin gene, gentamicin suppressed truncation of the protein and ameliorated the phenotype. A subset of patients with Duchenne and Becker muscular dystrophy similarly possess a nonsense mutation, causing premature termination of dystrophin translation. Four such patients, with various stop codon sequences, were treated once daily with intravenous gentamicin at 7.5 mg/kg/day for 2 weeks. No ototoxicity or nephrotoxicity was detected. Full-length dystrophin was not detected in pre- and post-treatment muscle biopsies. C1 Johns Hopkins Hosp, Dept Neurol, Baltimore, MD 21205 USA. NINCDS, Neurogenet Branch, NIH, Bethesda, MD 20892 USA. Childrens Natl Med Ctr, Med Genet Res Ctr, Washington, DC 20010 USA. NHGRI, Med Genet Branch, NIH, Bethesda, MD 20892 USA. NIH, Clin Pharmacokinet Res Lab, Bethesda, MD 20892 USA. George Washington Univ, Dept Neurol, Washington, DC USA. RP Wagner, KR (reprint author), Johns Hopkins Hosp, Dept Neurol, 607 PCTB,725 N Wolfe St, Baltimore, MD 21205 USA. FU NINDS NIH HHS [NS29525-09] NR 34 TC 153 Z9 157 U1 2 U2 10 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0364-5134 J9 ANN NEUROL JI Ann. Neurol. PD JUN PY 2001 VL 49 IS 6 BP 706 EP 711 DI 10.1002/ana.1023 PG 6 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 439BR UT WOS:000169091400005 PM 11409421 ER PT J AU Broderick, J Lu, M Jackson, C Pancioli, A Tilley, BC Fagan, SC Kothari, R Levine, SR Marler, JR Lyden, PD Haley, EC Brott, T Grotta, JC AF Broderick, J Lu, M Jackson, C Pancioli, A Tilley, BC Fagan, SC Kothari, R Levine, SR Marler, JR Lyden, PD Haley, EC Brott, T Grotta, JC CA NINDS t-PA Stroke Study Grp TI Apolipoprotein E phenotype and the efficacy of intravenous tissue plasminogen activator in acute ischemic stroke SO ANNALS OF NEUROLOGY LA English DT Article ID CEREBRAL AMYLOID ANGIOPATHY; ALZHEIMERS-DISEASE; E POLYMORPHISM; IN-VITRO; NEURITE OUTGROWTH; COGNITIVE DECLINE; EPSILON-2 ALLELE; BETA-PEPTIDE; HEAD-INJURY; E GENOTYPE AB We used stored plasma samples from 409 patients in the National Institute of Neurological Diseases and Stroke (NINDS) tissue plasminogen activator (t-PA) Stroke Trial to examine the relationship between an apolipoprotein (Apo) E2 or an Apo E4 phenotype and a favorable outcome 3 months after stroke, the risk of intracerebral hemorrhage, and the response to intravenous t-PA therapy. For the 27 patients with an Apo E2 phenotype who were treated with t-PA, the odds ratio (OR) of a favorable outcome at 3 months was 6.4 [95% confidence interval (CI) 2.7-15.3%] compared to the 161 patients without an Apo E2 phenotype who were treated with placebo. The 190 patients treated with t-PA who did not have an Apo E2 phenotype also had a greater, though less pronounced, likelihood of a favorable outcome (OR 2.0, 95% CI 1.2-3.2%) than patients without an Apo E2 phenotype treated with placebo. For the 31 patients with an Apo E2 phenotype treated with placebo, the OR of a favorable 3 month outcome was 0.8 (95% CI 0.4-1.7%) compared to the 161 patients without an Apo E2 phenotype treated with placebo. This interaction between treatment and Apo E2 status persisted after adjustment for baseline variables previously associated with 3 month outcome, for differences in the baseline variables in the two treatment groups and in the Apo E2-positive and -negative groups, and for a previously reported time-to-treatment x treatment interaction (p = 0.03). Apo E4 phenotype, present in 111 (27%) of the 409 patients, was not related to a favorable 3 month outcome, response to t-PA, 3 month mortality, or risk of intracerebral hemorrhage. We conclude that the efficacy of intravenous t-PA in patients with acute ischemic stroke may be enhanced in patients who have an Apo E2 phenotype, whereas the Apo E2 phenotype alone is not associated with a detectable benefit on stroke outcome at 3 months in patients not given t-PA. In contrast to prior studies of head injury and stroke, we could not detect a relationship between Apo E4 phenotype and clinical outcome. C1 Univ Cincinnati, Dept Neurol, Cincinnati, OH 45267 USA. Henry Ford Hlth Syst, Dept Biostat & Res Epidemiol, Detroit, MI USA. Univ Calif San Diego, Stroke Ctr, San Diego, CA 92103 USA. Univ Cincinnati, Dept Emergency Med, Cincinnati, OH USA. Med Univ S Carolina, Dept Biometry & Epidemiol, Charleston, SC 29425 USA. Univ Georgia, Augusta, GA USA. Borgess Res Inst, Kalamazoo, MI USA. Wayne State Univ, Stroke Program, Detroit, MI USA. NINDS, Div Stroke & Trauma, Bethesda, MD 20892 USA. Univ Virginia Hlth Syst, Dept Neurol, Charlottesville, VA USA. Mayo Clin, Dept Neurol, Jacksonville, FL 32224 USA. Univ Texas, Dept Neurol, Houston, TX 77225 USA. RP Broderick, J (reprint author), Univ Cincinnati, Dept Neurol, 231 Bethesda Ave,ML 0525, Cincinnati, OH 45267 USA. RI Fagan, Susan /D-3281-2009; Hill, Michael/C-9073-2012 OI Hill, Michael/0000-0002-6269-1543 FU NINDS NIH HHS [N01-NS-02374, N01-NS-02377, N01-NS-02382] NR 54 TC 32 Z9 32 U1 1 U2 1 PU WILEY-LISS PI NEW YORK PA DIV JOHN WILEY & SONS INC, 605 THIRD AVE, NEW YORK, NY 10158-0012 USA SN 0364-5134 J9 ANN NEUROL JI Ann. Neurol. PD JUN PY 2001 VL 49 IS 6 BP 736 EP 744 DI 10.1002/ana.1058 PG 9 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 439BR UT WOS:000169091400009 PM 11409425 ER PT J AU Gasch, AT Janjani, A Weber, M Garland, DL Chan, CC AF Gasch, AT Janjani, A Weber, M Garland, DL Chan, CC TI Ocular pathology and lens analysis in Hajdu-Cheney syndrome SO ANNALS OF OPHTHALMOLOGY LA English DT Article AB Ocular histopathologic and lens analyses were performed on the eyes of a M-year-old woman with Hajdu-Cheney syndrome. Compared with normal, age-matched lenses, the anterior cortical suture pattern was more complex and the crystallin composition was notably similar. Histologic findings included cataracts, multiple drusen, high myopia, and cystic retinal tuft The latter two findings can lead to retinal breaks. Periodic, comprehensive ophthalmologic examinations may he particularly important for individuals with Hajdu-Cheney syndrome. C1 NEI, NIH, Bethesda, MD 20892 USA. Ctr Hosp Univ Nantes, Nantes, France. RP Chan, CC (reprint author), NEI, NIH, 10 Ctr Dr,Bldg 10,Room 10N 103, Bethesda, MD 20892 USA. NR 16 TC 0 Z9 0 U1 0 U2 0 PU AMER SOC CONTEMPORARY OPHTHALMOLOGY PI CHICAGO PA 820 N ORLEANS, STE 208, CHICAGO, IL 60610 USA SN 1079-4794 J9 ANN OPHTHALMOL JI Ann. Ophthalmol. PD SUM PY 2001 VL 33 IS 2 BP 141 EP 144 DI 10.1007/s12009-001-0011-3 PG 4 WC Ophthalmology SC Ophthalmology GA 453ND UT WOS:000169921200016 ER PT J AU Schrump, DS AF Schrump, DS TI Efficient naked plasmid cotransfection of lung grafts by extended lung/plasmid exposure time - Invited commentary SO ANNALS OF THORACIC SURGERY LA English DT Editorial Material C1 NCI, Thorac Oncol Sect, Surg Branch, Bethesda, MD 20892 USA. RP Schrump, DS (reprint author), NCI, Thorac Oncol Sect, Surg Branch, 10 Ctr Dr,Room 2B07, Bethesda, MD 20892 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 0003-4975 J9 ANN THORAC SURG JI Ann. Thorac. Surg. PD JUN PY 2001 VL 71 IS 6 BP 1823 EP 1824 DI 10.1016/S0003-4975(01)02775-8 PG 2 WC Cardiac & Cardiovascular Systems; Respiratory System; Surgery SC Cardiovascular System & Cardiology; Respiratory System; Surgery GA 442QG UT WOS:000169296000018 ER PT J AU Espinel-Ingroff, A Bartlett, M Chaturvedi, V Ghannoum, M Hazen, KC Pfaller, MA Rinaldi, M Walsh, TJ AF Espinel-Ingroff, A Bartlett, M Chaturvedi, V Ghannoum, M Hazen, KC Pfaller, MA Rinaldi, M Walsh, TJ TI Optimal susceptibility testing conditions for detection of azole resistance in Aspergillus spp.: NCCLS collaborative evaluation SO ANTIMICROBIAL AGENTS AND CHEMOTHERAPY LA English DT Article ID IN-VITRO ACTIVITY; AMPHOTERICIN-B; FILAMENTOUS FUNGI; INVASIVE ASPERGILLOSIS; ANTIFUNGAL AGENTS; DIMORPHIC FUNGI; INOCULUM SIZE; ITRACONAZOLE; TERREUS; FUMIGATUS AB The most important role of susceptibility testing is to identify potentially resistant isolates for the agent being evaluated, Standard testing guidelines recently have been proposed for antifungal susceptibility testing of filamentous fungi (molds), This collaborative (eight centers) study evaluated further newly proposed guidelines (NCCLS, proposed standard M38-P, 1998) and other testing conditions for antifungal susceptibility testing of Aspergillus spp. to itraconazole and three new triazoles, posaconazole CSCH56592), ravuconazole (BMS-207147), and voriconazole. MICs of itraconazole, posaconazole, ravuconazole, and voriconazole for 15 selected isolates of three species of Aspergillus (A. fumigatus, A. flavus, and A. terreus) with well documented in vitro, clinical, or animal data were determined in each center by using four medium formulations (standard RPMI-1640 [RPMI], RPMI with 2% dextrose, antibiotic medium 3 [M3], and M3 with 2% dextrose) and two criteria of MIC determination (complete [MIC-0s] and prominent [MIC-2s] growth inhibition) at 24, 48, and 72 h, The highest reproducibility (92 to 99%) was seen dth the standard RPM[I and M3 media. Moreover, the distinction between itraconazole-resistant MICs of >8 mug/ml for clinically resistant strains) and -susceptible (MICs of 0.03 to 1 mug/ml) isolates, as well as between a voriconazole-resistant laboratory mutant and other isolates (voriconazole MICs of 2 to >8 versus 0.12 to 2 mug/ml), was more consistently evident with the standard RPMI medium and when MIG-Os were determined at 48 h, These results provide further refinement of the testing guidelines for susceptibility testing of Aspergillus spp. and warrant consideration for inclusion in the future NCCLS document M38-A. C1 Virginia Commonwealth Univ, Med Coll Virginia, Med Mycol Res Lab, Richmond, VA 23298 USA. Univ Virginia, Hlth Syst, Charlottesville, VA USA. Indiana Univ, Sch Med, Indianapolis, IN USA. Case Western Reserve Univ, Univ Hosp Cleveland, Cleveland, OH 44106 USA. New York Hlth Dept, Albany, NY USA. Univ Iowa, Coll Med, Iowa City, IA USA. Univ Texas, Hlth Sci Ctr, San Antonio, TX USA. NCI, Bethesda, MD 20892 USA. RP Espinel-Ingroff, A (reprint author), Virginia Commonwealth Univ, Med Coll Virginia, Med Mycol Res Lab, POB 980049,1101 E Marshall St,Sanger Hall,Room 7-, Richmond, VA 23298 USA. NR 31 TC 63 Z9 64 U1 0 U2 2 PU AMER SOC MICROBIOLOGY PI WASHINGTON PA 1752 N ST NW, WASHINGTON, DC 20036-2904 USA SN 0066-4804 J9 ANTIMICROB AGENTS CH JI Antimicrob. Agents Chemother. PD JUN PY 2001 VL 45 IS 6 BP 1828 EP 1835 DI 10.1128/AAC.45.6.1828-1835.2001 PG 8 WC Microbiology; Pharmacology & Pharmacy SC Microbiology; Pharmacology & Pharmacy GA 434YD UT WOS:000168842700033 PM 11353633 ER PT J AU Schmidt, AC Couch, RB Galasso, GJ Hayden, FG Mills, J Murphy, BR Chanock, RM AF Schmidt, AC Couch, RB Galasso, GJ Hayden, FG Mills, J Murphy, BR Chanock, RM TI Current research on respiratory viral infections: Third International Symposium SO ANTIVIRAL RESEARCH LA English DT Review ID SYNCYTIAL VIRUS-INFECTION; ACUTE OTITIS-MEDIA; INFLUENZA-A-VIRUS; NONTYPABLE HAEMOPHILUS-INFLUENZAE; POLYMERASE-CHAIN-REACTION; CONGENITAL HEART-DISEASE; ALPHA-INDUCED APOPTOSIS; COLD-ADAPTED INFLUENZA; RAPID DIAGNOSIS; YOUNG-CHILDREN C1 NIAID, Infect Dis Lab, NIH, Bethesda, MD 20892 USA. Baylor Coll Med, Dept Mol Virol & Microbiol, Houston, TX 77030 USA. Macrae Grp, Rockville, MD USA. Univ Virginia, Sch Med, Charlottesville, VA 22908 USA. Macfarlane Burnet Ctr Med Res, Melbourne, Vic, Australia. RP Schmidt, AC (reprint author), NIAID, Infect Dis Lab, NIH, 7 Ctr Dr, Bethesda, MD 20892 USA. NR 155 TC 21 Z9 22 U1 0 U2 0 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 0166-3542 J9 ANTIVIR RES JI Antiviral Res. PD JUN PY 2001 VL 50 IS 3 BP 157 EP 196 DI 10.1016/S0166-3542(01)00136-X PG 40 WC Pharmacology & Pharmacy; Virology SC Pharmacology & Pharmacy; Virology GA 450BC UT WOS:000169720800001 PM 11397506 ER PT J AU Phillips, AC Vousden, KH AF Phillips, AC Vousden, KH TI E2F-1 induced apoptosis SO APOPTOSIS LA English DT Review DE apoptosis; cell cycle; E2F1; p53 ID S-PHASE ENTRY; NF-KAPPA-B; TRANSCRIPTION FACTOR E2F-1; ARF TUMOR-SUPPRESSOR; THYMIC NEGATIVE SELECTION; SKELETAL-MUSCLE CELLS; RETINOBLASTOMA PROTEIN; HISTONE DEACETYLASE; IN-VIVO; P53-DEPENDENT APOPTOSIS AB Members of the E2F family of transcription factors play an important role in regulating the cell cycle, and their activity is often perturbed during the development of human malignancies. More recent work has shown that E2F-1 regulates apoptosis as well as proliferation, in part by stabilizing the p53 tumor suppressor, an important mediator of apoptosis. This has led to the suggestion that E2F-1 may function as a tumor surveillance mechanism, detecting aberrant proliferation and engaging apoptotic pathways to protect the organism from developing tumors. C1 NCI, Regulat Cell Growth Lab, FCRDC, Frederick, MD 21702 USA. RP Vousden, KH (reprint author), NCI, Regulat Cell Growth Lab, FCRDC, Bldg 560,Room 22-96,W 7th St, Frederick, MD 21702 USA. NR 112 TC 111 Z9 113 U1 0 U2 0 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 1360-8185 J9 APOPTOSIS JI Apoptosis PD JUN PY 2001 VL 6 IS 3 BP 173 EP 182 DI 10.1023/A:1011332625740 PG 10 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 417XM UT WOS:000167860800003 PM 11388666 ER PT J AU Danilkovitch-Miagkova, A Leonard, EJ AF Danilkovitch-Miagkova, A Leonard, EJ TI Anti-apoptotic action of macrophage stimulating protein (MSP) SO APOPTOSIS LA English DT Article DE adhesion; apoptosis; epithelial cells; MSP; signaling ID HEPATOCYTE GROWTH-FACTOR; RECEPTOR TYROSINE KINASE; FOCAL ADHESION KINASE; CELL-CYCLE PROGRESSION; FACTOR-I; EXTRACELLULAR-MATRIX; SIGNALING PATHWAYS; EPITHELIAL-CELLS; RON GENE; PHOSPHATIDYLINOSITOL-3 KINASE AB MSP is a serum protein belonging to the plasminogen-related kringle domain protein family. In addition to macrophages, epithelial cells are also MSP targets. MSP is a multifunctional factor regulating cell adhesion and motility, growth and survival. MSP mediates its biological activities by activating a transmembrane receptor tyrosine kinase called RON in humans or SKT in mice. MSP can protect epithelial cells from apoptosis by activating two independent signals in the PI3-K/AKT or the MAPK pathway. The MAPK pathway mediates the MSP anti-apoptotic effect only if additional signaling pathways are activated through adhesion. This indicates that MSP receptors and integrins, the receptors mediating cell-matrix-dependent adhesion, can collaborate in promotion of cell survival. This adhesion-dependent pathway, which is essential for the MAPK-mediated anti-apoptotic effect, remains to be identified. A hypothesis that Stat3 might represent a key component of the adhesion-induced anti-apoptotic pathway is presented in this review. C1 NCI, Immunopathol Sect, Immunobiol Lab, Frederick, MD 21702 USA. RP Danilkovitch-Miagkova, A (reprint author), NCI, Immunopathol Sect, Immunobiol Lab, Bldg 560,Room 1246, Frederick, MD 21702 USA. NR 82 TC 25 Z9 25 U1 0 U2 0 PU KLUWER ACADEMIC PUBL PI DORDRECHT PA SPUIBOULEVARD 50, PO BOX 17, 3300 AA DORDRECHT, NETHERLANDS SN 1360-8185 J9 APOPTOSIS JI Apoptosis PD JUN PY 2001 VL 6 IS 3 BP 183 EP 190 DI 10.1023/A:1011384609811 PG 8 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 417XM UT WOS:000167860800004 PM 11388667 ER PT J AU Goddard, AW Mason, GF Almai, A Rothman, DL Behar, KL Petroff, OAC Charney, DS Krystal, JH AF Goddard, AW Mason, GF Almai, A Rothman, DL Behar, KL Petroff, OAC Charney, DS Krystal, JH TI Reductions in occipital cortex GABA levels in panic disorder detected with H-1-magnetic resonance spectroscopy SO ARCHIVES OF GENERAL PSYCHIATRY LA English DT Article ID GAMMA-AMINOBUTYRIC-ACID; BENZODIAZEPINE RECEPTOR-BINDING; BRAIN GABA; IOMAZENIL-SPECT; CORTICAL GABA; HOMOCARNOSINE; VIGABATRIN; EPILEPSY; ANXIETY; MICE AB Background: There is preclinical evidence and indirect clinical evidence implicating gamma -aminobutyric acid (GABA) in the pathophysiology and treatment of human panic disorder. Specifically, deficits in GABA neuronal function have been associated with anxiogenesis, whereas enhancement of GABA function tends to be anxiolytic. Although reported peripheral GABA levels leg, in cerebrospinal fluid and plasma) have been within reference limits in panic disorder, thus far there has been no direct assessment of brain GABA levels in this disorder. The purpose of the present work was to determine whether cortical GABA levels are abnormally low in patients with panic disorder. Methods: Total occipital cortical GABA levels (GABA plus homocarnosine) were assessed in 14 unmedicated patients with panic disorder who did not have major depression and 14 retrospectively age- and sex-marched control subjects using spatially localized H-1-magnetic reso-nance spectroscopy. All patients met DSM-IV criteria for a principal current diagnosis of panic disorder with or without agoraphobia. Results: Patients with panic disorder had a 22% reduction in total occipital cortex GABA concentration (GABA plus homocarnosine) compared with controls. This finding was present in 12 of 14 patient-control pairs and was not solely accounted for by medication history. There were no significant correlations between occipital cortex GABA levels and measures of illness or state anxiety. Conclusions: Panic disorder is associated with reductions in total occipital cortex GABA levels. This abnormality might contribute to the pathophysiology, of panic disorder. C1 Yale Univ, Dept Psychiat, Yale Anxiety Clin, New Haven, CT 06511 USA. Yale Univ, Dept Biomed Engn, New Haven, CT 06511 USA. Yale Univ, Dept Internal Med, New Haven, CT 06511 USA. Yale Univ, Dept Radiol, New Haven, CT 06511 USA. Yale Univ, Dept Neurol, New Haven, CT 06511 USA. NIMH, Rockville, MD 20857 USA. RP Goddard, AW (reprint author), Yale Univ, Dept Psychiat, Yale Anxiety Clin, 100 York St,Room 2J, New Haven, CT 06511 USA. FU NIAAA NIH HHS [K02 1 AA00261-01, K02 AA000261]; NIMH NIH HHS [K-08 MH-01322, MH-30929, R01 MH-58657]; NINDS NIH HHS [R01 NS034813, R01-NS34813]; PHS HHS [R01-N53218, R29-N5032126] NR 42 TC 145 Z9 152 U1 1 U2 14 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0003-990X J9 ARCH GEN PSYCHIAT JI Arch. Gen. Psychiatry PD JUN PY 2001 VL 58 IS 6 BP 556 EP 561 DI 10.1001/archpsyc.58.6.556 PG 6 WC Psychiatry SC Psychiatry GA 439UT UT WOS:000169136500005 PM 11386984 ER PT J AU Schiffman, RM Jacobsen, G Whitcup, SM AF Schiffman, RM Jacobsen, G Whitcup, SM TI Visual functioning and general health status in patients with uveitis SO ARCHIVES OF OPHTHALMOLOGY LA English DT Article ID QUALITY-OF-LIFE; CATARACT; ACUITY; INDEX; VF-14 AB Objective: To measure the visual functioning and quality of life in patients with uveitis. Methods: Consecutive adult patients with noninfectious uveitis were enrolled. The Medical Outcomes Study 36-Item Short Form (SF-36) and the National Eye Institute Visual Functioning Questionnaire (NEI VFQ-25) were administered by a trained interviewer. Sociodemographic and clinical data were also collected. Results: Seventy-six patients were enrolled. The overall NEI VFQ-25 score was significantly lower among patients with uveitis than in a normal reference group (P < .001). The SF-36 physical (PCS) and mental (MCS) component summary scores were also significantly lower among patients with uveitis than in the general US population (P < .007). Among patients with uveitis, visual acuity, binocular involvement, intensity of therapy, employment status, and PCS and MCS scores were all significantly associated with overall NEI VFQ-ZS scores in multivariable analysis. Medical comorbidity,ocular comorbidity, and NEI VFQ-25 scores were significantly associated with PCS scores. Medical comorbidity and NEI VFQ-25 scores were significantly associated with MCS scores. Regression models including NEI VFQ-ZS scores explained an additional 7% of the variance in PCS scores and 16% of MCS scores. Models including both PCS and MCS scores explained an additional 12% of the variance in NEI VFQ-2S scores. Conclusions: Patients with uveitis reported markedly poorer visual functioning and general health status than normal subjects. Patients with more severe uveitis have poorer visual functioning and general health status than patients with milder disease. Visual functioning-and general health status measurement contribute complementary information and should both be performed in patients with uveitis to measure the effect of disease and its therapy on their quality of life. C1 Henry Ford Hlth Syst, Eye Care Serv K10, Dept Eye Care Serv, Detroit, MI 48202 USA. Henry Ford Hlth Syst, Dept Biostat & Res Epidemiol, Detroit, MI 48202 USA. NEI, Clin Branch, Bethesda, MD 20892 USA. RP Schiffman, RM (reprint author), Henry Ford Hlth Syst, Eye Care Serv K10, Dept Eye Care Serv, 2799 W Grand Blvd, Detroit, MI 48202 USA. NR 17 TC 65 Z9 66 U1 0 U2 2 PU AMER MEDICAL ASSOC PI CHICAGO PA 515 N STATE ST, CHICAGO, IL 60610 USA SN 0003-9950 J9 ARCH OPHTHALMOL-CHIC JI Arch. Ophthalmol. PD JUN PY 2001 VL 119 IS 6 BP 841 EP 849 PG 9 WC Ophthalmology SC Ophthalmology GA 440JC UT WOS:000169171300006 PM 11405835 ER PT J AU Smith, JR Levinson, RD Holland, GN Jabs, DA Robinson, MR Whitcup, SM Rosenbaum, JT AF Smith, JR Levinson, RD Holland, GN Jabs, DA Robinson, MR Whitcup, SM Rosenbaum, JT TI Differential efficacy of tumor necrosis factor inhibition in the management of inflammatory eye disease and associated rheumatic disease SO ARTHRITIS & RHEUMATISM-ARTHRITIS CARE & RESEARCH LA English DT Article ID ENDOTOXIN-INDUCED UVEITIS; CHIMERIC MONOCLONAL-ANTIBODY; FACTOR-ALPHA; TNF-ALPHA; ARTHRITIS; METHOTREXATE; ETANERCEPT; INFLIXIMAB; TRIAL; RAT AB Objective. To evaluate the clinical usefulness of tumor necrosis factor (TNF) inhibitors in patients with inflammatory eye disease that is resistant to conventional immunosuppressive therapies. Methods. Sixteen patients (4 males and 12 females aged 7 to 78 years) who received etanercept (n = 14) or infliximab (n 2) for either inflammatory eye disease or associated joint disease were studied retrospectively to determine the effect of these medications on their ocular inflammation. Results. Nine cases of uveitis and 7 cases of scleritis were treated. Systemic diagnoses included rheumatoid arthritis (n 8), juvenile rheumatoid arthritis (n = 3), ankylosing spondylitis (n = 1), and psoriatic spondylarthropathy (n = 1). Three patients had uveitis without associated systemic disease. Although 12 of 12 patients with active articular inflammation (100%) experienced improvement in joint disease, only 6 of 16 with ocular inflammation (38%) experienced improvement in eye disease. Five patients developed inflammatory eye disease for the first time while taking a TNF inhibitor. No patient discontinued treatment because of adverse drug effects. Conclusion. TNF inhibitors are well tolerated immunosuppressive medications that may benefit certain subgroups of patients with inflammatory eye disease, but they appear to be more effective in controlling associated inflammatory arthritis. C1 Oregon Hlth Sci Univ, Casey Eye Inst, Portland, OR 97201 USA. Univ Calif Los Angeles, Sch Med, UCLA Ocular Inflammatory Dis Ctr, Dept Ophthalmol, Los Angeles, CA 90024 USA. Univ Calif Los Angeles, Sch Med, Jules Stein Eye Inst, Los Angeles, CA 90024 USA. Johns Hopkins Univ, Sch Med, Wilmer Eye Inst, Dept Ophthalmol, Baltimore, MD 21205 USA. Johns Hopkins Univ, Sch Med, Dept Med, Baltimore, MD 21205 USA. NEI, NIH, Bethesda, MD 20892 USA. RP Smith, JR (reprint author), Oregon Hlth Sci Univ, Casey Eye Inst, 3375 Terwilliger Blvd, Portland, OR 97201 USA. FU NEI NIH HHS [EY-06484] NR 21 TC 204 Z9 211 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 0004-3591 J9 ARTHRIT RHEUM-ARTHR JI Arthritis Rheum-Arthritis Care Res. PD JUN PY 2001 VL 45 IS 3 BP 252 EP 257 DI 10.1002/1529-0131(200106)45:3<252::AID-ART257>3.0.CO;2-5 PG 6 WC Rheumatology SC Rheumatology GA 487JT UT WOS:000171872500008 PM 11409666 ER PT J AU Weisberg, IS Jacques, PF Selhub, J Bostom, AG Chen, ZT Ellison, RC Eckfeldt, JH Rozen, R AF Weisberg, IS Jacques, PF Selhub, J Bostom, AG Chen, ZT Ellison, RC Eckfeldt, JH Rozen, R TI The 129XA -> C polymorphism in methylenetetrahydrofolate reductase (MTHFR): in vitro expression and association with homocysteine SO ATHEROSCLEROSIS LA English DT Article DE homocysteine; folate; genetics; enzymes; methylenetetrahydrofalate reductase ID CORONARY-ARTERY DISEASE; GENETIC RISK FACTOR; COMMON MUTATION; PLASMA HOMOCYSTEINE; VASCULAR-DISEASE; HYPERHOMOCYSTEINEMIA AB A common mutation in methylenetetrahydrofolate reductase (MTHFR), 677C --> T, is associated with reduced enzyme activity, a thermolabile enzyme and mild hyperhomocysteinemia, a risk factor for vascular disease. Recently, a second common mutation (1298A --> C; glutamate to alanine) was reported, but this mutation was suggested to increase homocysteine only in individuals who carried the bp677 variant. To evaluate the functional consequences of this mutation, we performed site-directed mutagenesis and in vitro expression. For in vivo assessment of clinical impact, we examined the 1298A --> C genotypes and plasma homocysteine in 198 individuals from the NHLBI Family Heart Study that had previously been assessed for the 677 substitution. Site-directed mutagenesis of the human cDNA was performed to generate enzymes containing each of the two mutations, as well as an enzyme containing both substitutions. Enzyme activity and thermolability were assessed in bacterial extracts. The activity of the wild-type cDNA was designated as 100%; mutant enzymes containing the 1298 and 677 mutations separately had 68% (+/- 5.0) and 45% (+/- 10.8), respectively, of control activity while the enzyme containing both mutations had 41% (+/- 12.8) of control activity. The 1298 mutation was not associated with a thermolabile enzyme. In the Family Heart Study, fasting homocysteine was significantly higher (P < 0.05) in individuals heterozygous for both substitutions, compared to individuals who carried only the 677C --> T variant. This study suggests that two variants in MTHFR should be assessed as genetic risk factors for hyperhomocysteinemia. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved. C1 McGill Univ, Ctr Hlth, Dept Human Genet, Montreal, PQ, Canada. McGill Univ, Ctr Hlth, Dept Pediat, Montreal, PQ, Canada. Tufts Univ, Jean Mayer USDA Human Nutr Res Ctr Aging, Boston, MA 02111 USA. Mem Hosp Rhode Isl, Div Gen Internal Med, Providence, RI USA. Boston Univ, Sch Med, Boston, MA 02215 USA. NHLBI, Family Heart Study Field Ctr, Framingham, MA USA. Univ Minnesota, Dept Lab Med & Pathol, NHLBI, Family Heart Study Cent Lab, Minnesota, MN USA. RP Rozen, R (reprint author), Montreal Childrens Hosp, 4060 St Catherine W, Montreal, PQ H3Z 2Z3, Canada. FU NHLBI NIH HHS [N01-HC-25106] NR 25 TC 252 Z9 268 U1 0 U2 3 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 JUN PY 2001 VL 156 IS 2 BP 409 EP 415 DI 10.1016/S0021-9150(00)00671-7 PG 7 WC Cardiac & Cardiovascular Systems; Peripheral Vascular Disease SC Cardiovascular System & Cardiology GA 445UJ UT WOS:000169476700019 PM 11395038 ER PT J AU Rapoport, JL Castellanos, FX Gogate, N Janson, K Kohler, S Nelson, P AF Rapoport, JL Castellanos, FX Gogate, N Janson, K Kohler, S Nelson, P TI Imaging normal and abnormal brain development: new perspectives for child psychiatry SO AUSTRALIAN AND NEW ZEALAND JOURNAL OF PSYCHIATRY LA English DT Article DE ADHD; brain development; childhood schizophrenia ID DEFICIT HYPERACTIVITY DISORDER; ATTENTION-DEFICIT/HYPERACTIVITY DISORDER; OCULAR DOMINANCE COLUMNS; ONSET SCHIZOPHRENIA; SYNAPSE ELIMINATION; CAUDATE-NUCLEUS; IN-VIVO; NEUROMUSCULAR-JUNCTION; DOPAMINERGIC-NEURONS; PREFRONTAL CORTEX AB Objective: The availability of non-invasive brain imaging permits the study of normal and abnormal brain development in childhood and adolescence. This paper summarizes current knowledge of brain abnormalities of two conditions, attention deficit hyperactivity disorder (ADHD) and childhood onset schizophrenia (COS), and illustrates how such findings are bringing clinical and preclinical perspectives closer together. Method: A selected review is presented of the pattern and temporal characteristics of anatomic brain magnetic resonance imaging (MRI) studies in ADHD and COS. These results are discussed in terms of candidate mechanisms suggested by studies in developmental neuroscience. Results: There are consistent, diagnostically specific patterns of brain abnormality for ADHD and COS. Attention deficit hyperactivity disorder is characterized by a slightly smaller (4%) total brain volume (both white and grey matter), less-consistent abnormalities of the basal ganglia and a striking (15%) decrease in posterior inferior cerebellar vermal volume. These changes do not progress with age. In contrast, patients with COS have smaller brain volume due to a 10% decrease in cortical grey volume. Moreover, in COS there is a progressive loss of regional grey volume particularly in frontal and temporal regions during adolescence. Conclusions: In ADHD, the developmental pattern suggests an early non-progressive 'lesion' involving neurotrophic factors controlling overall brain growth and selected dopamine circuits. In contrast, in COS, which shows progressive grey matter loss, various candidate processes influencing later synaptic and dendritic pruning are suggested by human post-mortem and developmental animal studies. C1 NIMH, Child Psychiat Branch, Bethesda, MD 20892 USA. NICHHD, Lab Dev Neurobiol Branch, Neurobiol Sect, Bethesda, MD 20892 USA. RP Rapoport, JL (reprint author), NIMH, Child Psychiat Branch, Bldg 10,Room 3N202,10 Ctr Dr MSC 1600, Bethesda, MD 20892 USA. RI Gogtay, Nitin/A-3035-2008 NR 78 TC 48 Z9 50 U1 5 U2 8 PU BLACKWELL SCIENCE ASIA PI CARLTON PA 54 UNIVERSITY ST, P O BOX 378, CARLTON, VICTORIA 3053, AUSTRALIA SN 0004-8674 J9 AUST NZ J PSYCHIAT JI Aust. N. Z. J. Psych. PD JUN PY 2001 VL 35 IS 3 BP 272 EP 281 DI 10.1046/j.1440-1614.2001.00900.x PG 10 WC Psychiatry SC Psychiatry GA 462HW UT WOS:000170415500003 PM 11437799 ER PT J AU Bachevalier, J Malkova, L Mishkin, M AF Bachevalier, J Malkova, L Mishkin, M TI Effects of selective neonatal temporal lobe lesions on socioemotional behavior in infant rhesus monkeys (Macaca mulatta) SO BEHAVIORAL NEUROSCIENCE LA English DT Article ID KLUVER-BUCY SYNDROME; HIPPOCAMPAL DAMAGE; RECOGNITION MEMORY; VISUAL RECOGNITION; SOCIAL-BEHAVIOR; LIMBIC LESIONS; AUTISM; AMYGDALA; CHILDREN; CORTEX AB Normal infant monkeys and infant monkeys with neonatal damage to either the medial temporal lobe or the inferior temporal visual area were assessed in dyadic social interactions at 2 and 6 months of age, Unlike the normal infant monkeys, which developed strong affiliative bonds and little or no behavioral disturbances, the lesioned monkeys (each of which was observed with an unoperated control) exhibited socioemotional abnormalities and aberrant behaviors. The socioemotional changes predominated at 6 months of age and were particularly severe in monkeys with medial temporal lesions. In both the pattern and time course, the socioemotional deficits produced by the neonatal medial temporal lesions bear a striking resemblance to the behavioral syndrome in children with autism. Further analysis of these lesion-induced abnormalities in nonhuman primates may therefore provide insight into this debilitating human developmental disorder. C1 NIMH, Neuropsychol Lab, Bethesda, MD 20892 USA. RP Bachevalier, J (reprint author), Univ Texas, Hlth Sci Ctr, Dept Neurobiol & Anat, 6431 Fannin, Houston, TX 77030 USA. FU NICHD NIH HHS [HD 35471]; PHS HHS [58846] NR 88 TC 43 Z9 44 U1 1 U2 3 PU AMER PSYCHOLOGICAL ASSOC PI WASHINGTON PA 750 FIRST ST NE, WASHINGTON, DC 20002-4242 USA SN 0735-7044 J9 BEHAV NEUROSCI JI Behav. Neurosci. PD JUN PY 2001 VL 115 IS 3 BP 545 EP 559 DI 10.1037//0735-7044.115.3.545 PG 15 WC Behavioral Sciences; Neurosciences SC Behavioral Sciences; Neurosciences & Neurology GA 471DC UT WOS:000170911500002 PM 11439445 ER PT J AU Langford, CA AF Langford, CA TI Management of systemic vasculitis SO BEST PRACTICE & RESEARCH IN CLINICAL RHEUMATOLOGY LA English DT Article DE vasculitis; arteritis; Wegener; treatment; toxicity; glucocorticoid; cytotoxic; cyclophosphamide; methotrexate ID GIANT-CELL ARTERITIS; CHURG-STRAUSS-SYNDROME; WEGENERS-GRANULOMATOSIS; POLYARTERITIS-NODOSA; TAKAYASU-ARTERITIS; MICROSCOPIC POLYANGIITIS; PULSE CYCLOPHOSPHAMIDE; MYCOPHENOLATE-MOFETIL; CLINICAL-FEATURES; RANDOMIZED TRIAL AB The systemic vasculitides are a wide-ranging group of diseases that are characterized by the presence of blood vessel inflammation. Despite this common feature, each type of vasculitis has a unique variety of clinical manifestations that influences its degree of disease severity and ultimately its management. Immunosuppressive therapy forms the foundation of treatment for almost all forms of systemic vasculitis. Because of this, treatment can be associated with its own risk of morbidity, or even mortality, related to specific medication side-effects or infections which occur as a result of impaired host defences. This chapter seeks to review the approach to management in selected forms of systemic vasculitis. Questions examined include the following. When should one treat systemic vasculitis! How does the nature of the disease and its severity affect treatment decisions! What are the data regarding the effectiveness of individual therapeutic regimens?. C1 NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. RP Langford, CA (reprint author), NIAID, Immunoregulat Lab, NIH, Bethesda, MD 20892 USA. NR 51 TC 14 Z9 14 U1 0 U2 2 PU BAILLIERE TINDALL PI LONDON PA 24-28 OVAL RD, LONDON NW1 7DX, ENGLAND SN 1521-6942 J9 BEST PRACT RES CL RH JI Best Pract. Res. Clin. Rheumatol. PD JUN PY 2001 VL 15 IS 2 BP 281 EP 297 DI 10.1053/berh.2001.0144 PG 17 WC Rheumatology SC Rheumatology GA 440TM UT WOS:000169190400007 PM 11469822 ER PT J AU Dundr, M Misteli, T AF Dundr, M Misteli, T TI Functional architecture in the cell nucleus SO BIOCHEMICAL JOURNAL LA English DT Review DE chromatin; gene expression; nuclear compartments; protein dynamics ID RNA-POLYMERASE-II; PRE-MESSENGER-RNA; RIBOSOMAL GENE-TRANSCRIPTION; SATELLITE DNA-SEQUENCES; PAIR-RULE TRANSCRIPTS; IN-SITU HYBRIDIZATION; SMALL NUCLEOLAR RNAS; LIVING HUMAN-CELLS; COILED BODIES; SACCHAROMYCES-CEREVISIAE AB The major functions of the cell nucleus, including transcription, pre-mRNA splicing and ribosome assembly, have been studied extensively by biochemical, genetic and molecular methods. An overwhelming amount of information about their molecular mechanisms is available. In stark contrast, very little is known about how these processes are integrated into the structural framework of the cell nucleus and how they are spatially and temporally co-ordinated within the three-dimensional confines of the nucleus. It is also largely unknown how nuclear architecture affects gene expression. In order to understand how genomes are organized, and how they function, the basic principles that govern nuclear architecture and function must be uncovered. Recent work combining molecular, biochemical and cell biological methods is beginning to shed light on how the nucleus functions and how genes are expressed in vivo. It has become clear that the nucleus contains distinct compartments and that many nuclear components are highly dynamic. Here we describe the major structural compartments of the cell nucleus and discuss their established and proposed functions. We summarize recent observations regarding the dynamic properties of chromatin, mRNA and nuclear proteins, and we consider the implications these findings have for the organization of nuclear processes and gene expression. Finally, we speculate that self-organization might play a substantial role in establishing and maintaining nuclear organization. C1 NCI, NIH, Bethesda, MD 20892 USA. RP Misteli, T (reprint author), NCI, NIH, 41 Lib Dr,Bldg 41, Bethesda, MD 20892 USA. NR 188 TC 270 Z9 281 U1 0 U2 13 PU PORTLAND PRESS PI LONDON PA 59 PORTLAND PLACE, LONDON W1N 3AJ, ENGLAND SN 0264-6021 J9 BIOCHEM J JI Biochem. J. PD JUN 1 PY 2001 VL 356 BP 297 EP 310 DI 10.1042/0264-6021:3560297 PN 2 PG 14 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 442QE UT WOS:000169295800001 PM 11368755 ER PT J AU Tanaka, T de la Concepcion, MLR De Luca, LM AF Tanaka, T de la Concepcion, MLR De Luca, LM TI Involvement of all-trans-retinoic acid in the breakdown of retinoic acid receptors alpha and gamma through proteasomes in MCF-7 human breast cancer cells SO BIOCHEMICAL PHARMACOLOGY LA English DT Article DE retinoic acid receptor; proteasomes; ubiquitination; retinoic acid; proteolysis ID SUMO-1 MODIFICATION; RESPONSE ELEMENT; VITAMIN-A; CYCLIN D1; PROTEINS; DIFFERENTIATION; INHIBITION; DEGRADATION; PROGRESSION; PROTEOLYSIS AB Most studies have reported an up-regulation of retinoic acid receptor (RAR) mRNA expression by all-trans retinoic acid (RA). We aimed to study the effect of RA on RAR protein levels in MCF-7 human breast cancer cells. Incubation of these cells with 10(-6) M RA induced a rapid breakdown of both RAR alpha and RAR gamma in spite of the accumulation of their mRNAs. Proteasome specific inhibitors blocked the RA-induced breakdown of RARs. Furthermore, RA enhanced the formation of the complex between RARa: and ubiquitin in a concentration and time-dependent manner, suggesting the involvement of ubiquitin and proteasome in this reaction. Retinoid X receptor cu (RXR alpha) was also decreased, albeit to a lesser extent, in RA-treated cells. Use of synthetic receptor agonists and antagonists clearly showed that the effect of the retinoid on the breakdown of the retinoid receptors is receptor-ligand agonist-dependent and blunted by the antagonist. An electrophoretic mobility shift assay, using nuclear extracts from RA-treated cells, showed that a reduction in complex formation with hormone response elements correlated with the reduction of RAR and RXR protein. These data suggest that RA induces the breakdown of RARs through a process involving ubiquitination and that this phenomenon causes a reduction in the formation of DNA-receptor complexes. (C) 2001 Elsevier Science Inc. All rights reserved. C1 NCI, NIH, Bethesda, MD 20892 USA. RP De Luca, LM (reprint author), NCI, NIH, Bldg 37,Room 3A-17,37 Convent Dr, Bethesda, MD 20892 USA. NR 30 TC 48 Z9 50 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 JUN 1 PY 2001 VL 61 IS 11 BP 1347 EP 1355 DI 10.1016/S0006-2952(01)00600-1 PG 9 WC Pharmacology & Pharmacy SC Pharmacology & Pharmacy GA 426CF UT WOS:000168331700003 PM 11331070 ER PT J AU Bushel, PR Hamadeh, H Bennett, L Sieber, S Martin, K Nuwaysir, EF Johnson, K Reynolds, K Paules, RS Afshari, CA AF Bushel, PR Hamadeh, H Bennett, L Sieber, S Martin, K Nuwaysir, EF Johnson, K Reynolds, K Paules, RS Afshari, CA TI MAPS: a microarray project system for gene expression experiment information and data validation SO BIOINFORMATICS LA English DT Article AB MAPS is a MicroArray Project System for management and interpretation of microarray gene expression experiment information and data. Microarray project information is organized to track experiments and results that are: (1) validated by performing analysis on stored replicate gene expression data; and (2) queried according to the biological classifications of genes deposited on microarray chips. C1 NIEHS, Mol Carcinogenesis Lab, Res Triangle Pk, NC 27709 USA. NIEHS, Lab Environm Carcinogenesis Mutagenesis, Res Triangle Pk, NC 27709 USA. RP Afshari, CA (reprint author), NIEHS, Mol Carcinogenesis Lab, POB 12233, Res Triangle Pk, NC 27709 USA. NR 5 TC 44 Z9 45 U1 0 U2 0 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 1367-4803 J9 BIOINFORMATICS JI Bioinformatics PD JUN PY 2001 VL 17 IS 6 BP 564 EP 565 DI 10.1093/bioinformatics/17.6.564 PG 2 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 444NC UT WOS:000169404700013 PM 11395436 ER PT J AU Sohl, G Eiberger, J Jung, YT Kozak, CA Willecke, K AF Sohl, G Eiberger, J Jung, YT Kozak, CA Willecke, K TI The mouse gap junction gene connexin29 is highly expressed in sciatic nerve and regulated during brain development SO BIOLOGICAL CHEMISTRY LA English DT Article DE connexin subgroup; Cx29; human Cx30.2; oligodendrocytes; peripheral nerves; spinal cord ID MARIE-TOOTH-DISEASE; MOLECULAR-CLONING; FUNCTIONAL EXPRESSION; RAT-BRAIN; PROTEIN; CELLS; OLIGODENDROCYTES; ASTROCYTES; MYELIN; FAMILY AB A novel mouse gap junction gene, coding for a presumptive protein of 258 amino acids (molecular mass: 28 981 Da), has been designated connexin29. This single copy gene was mapped to distal mouse chromosome 5 and shows 75% sequence identity to a human connexin30.2 sequence in the database. Connexin29 mRNA (4.4 kb) is highly expressed in mouse sciatic nerve and less abundant in spinal cord as well as in adult brain, where it increased 12-fold between day 7 and 14 post partum. Our expression data suggest that the new connexin gene is active in myelin-forming glial cells. C1 Univ Bonn, Inst Genet, Abt Mol Genet, D-53117 Bonn, Germany. NIAID, Mol Microbiol Lab, NIH, Bethesda, MD 20892 USA. RP Willecke, K (reprint author), Univ Bonn, Inst Genet, Abt Mol Genet, D-53117 Bonn, Germany. NR 38 TC 42 Z9 44 U1 0 U2 0 PU WALTER DE GRUYTER & CO PI BERLIN PA GENTHINER STRASSE 13, D-10785 BERLIN, GERMANY SN 1431-6730 J9 BIOL CHEM JI Biol. Chem. PD JUN PY 2001 VL 382 IS 6 BP 973 EP 978 DI 10.1515/BC.2001.122 PG 6 WC Biochemistry & Molecular Biology SC Biochemistry & Molecular Biology GA 457XN UT WOS:000170163000012 PM 11501764 ER PT J AU Baker, SG AF Baker, SG TI Discussion of double sampling for survival analysis SO BIOMETRICS LA English DT Editorial Material C1 NCI, Biometry Res Grp, Div Canc Prevent, Bethesda, MD 20892 USA. RP Baker, SG (reprint author), NCI, Biometry Res Grp, Div Canc Prevent, EPN 344,6130 Execut Blvd,MSC 7354, Bethesda, MD 20892 USA. NR 3 TC 1 Z9 1 U1 0 U2 0 PU INTERNATIONAL BIOMETRIC SOC PI WASHINGTON PA 1441 I ST, NW, SUITE 700, WASHINGTON, DC 20005-2210 USA SN 0006-341X J9 BIOMETRICS JI Biometrics PD JUN PY 2001 VL 57 IS 2 BP 348 EP 350 DI 10.1111/j.0006-341X.2001.00348.x PG 3 WC Biology; Mathematical & Computational Biology; Statistics & Probability SC Life Sciences & Biomedicine - Other Topics; Mathematical & Computational Biology; Mathematics GA 437QY UT WOS:000169006100003 PM 11414555 ER PT J AU Pinsky, PF AF Pinsky, PF TI Estimation and prediction for cancer screening models using deconvolution and smoothing SO BIOMETRICS LA English DT Article DE cancer screening; competing mortality; convolution model; deconvolution; lead time; overdiagnosis; sojourn time ID MEAN SOJOURN TIME; COLORECTAL-CANCER; SENSITIVITY; PROGRAMS; DISEASE; PHASE AB The model that specifies that cancer incidence, I, is the convolution of the preclinical incidence, g, and the density of time in the preclinical phase, f, has frequently been utilized to model data from cancer screening trials and to estimate such quantities as sojourn time, lead time, and sensitivity. When this model is fit to the above data, the parameters of f as well as the parameter(s) governing screening sensitivity must be estimated. Previously, g was either assumed to be equal to clinical incidence or assumed to be a constant or exponential function that also had to be estimated. Here we assume that the underlying incidence, I. in the study population (in the absence of screening) is known. With I known, g then becomes a function of f, which can be solved for using (numerical) deconvolution, thus eliminating the need to estimate g or make assumptions about it. Since numerical deconvolution procedures may be highly unstable, however, we incorporate a smoothing procedure that produces a realistic g function while still closely reproducing the original incidence function I upon convolution with f. We have also added the concept of competing mortality to the convolution model. This, along with the realistic preclinical incidence function described above, results in more accurate estimates of sojourn time and lead time and allows for estimation of quantities related to overdiagnosis, which we define here. C1 NCI, Div Canc Prevent, Bethesda, MD 20892 USA. RP Pinsky, PF (reprint author), NCI, Div Canc Prevent, EPN 330,6130 Execut Blvd, Bethesda, MD 20892 USA. NR 11 TC 18 Z9 18 U1 1 U2 5 PU INTERNATIONAL BIOMETRIC SOC PI WASHINGTON PA 1441 I ST, NW, SUITE 700, WASHINGTON, DC 20005-2210 USA SN 0006-341X J9 BIOMETRICS JI Biometrics PD JUN PY 2001 VL 57 IS 2 BP 389 EP 395 DI 10.1111/j.0006-341X.2001.00389.x PG 7 WC Biology; Mathematical & Computational Biology; Statistics & Probability SC Life Sciences & Biomedicine - Other Topics; Mathematical & Computational Biology; Mathematics GA 437QY UT WOS:000169006100010 PM 11414561 ER PT J AU Dunson, DB Baird, DD AF Dunson, DB Baird, DD TI A flexible parametric model for combining current status and age at first diagnosis data SO BIOMETRICS LA English DT Article DE coarse data; informative missingness; interval censoring; latency; proportional odds; survival analysis; three-state model; uterine fibroids ID NONPARAMETRIC-ESTIMATION; INCUBATION PERIOD; CENSORED-DATA; COARSE DATA; AIDS; INFECTION; IGNORABILITY; REGRESSION AB In some cross-sectional studies of chronic disease. data consist of the age at examination, whether the disease was present at the exam and recall of the age at first diagnosis. This article describes a flexible parametric approach for combining current status and age at first diagnosis data. We assume that the log odds of onset by a given age and of detection by a given age conditional on onset by that age are nondecreasing functions of time plus linear combinations of covariates. Piecewise linear models are used to characterize changes across time in the baseline odds. Methods are described for accommodating informatively missing current status data and inferences based on the age-specific incidence of disease prior to a landmark event (e.g., puberty, menopause). Our formulation enables straightforward maximum likelihood estimation without requiring restrictive parametric or Markov assumptions. The methods are applied to data from a study of uterine fibroids. C1 NIEHS, Biostat Branch, Res Triangle Pk, NC 27709 USA. NIEHS, Epidemiol Branch, Res Triangle Pk, NC 27709 USA. RP Dunson, DB (reprint author), NIEHS, Biostat Branch, MD A3-03,POB 12233, Res Triangle Pk, NC 27709 USA. OI Baird, Donna/0000-0002-5544-2653 NR 24 TC 10 Z9 10 U1 0 U2 2 PU INTERNATIONAL BIOMETRIC SOC PI WASHINGTON PA 1441 I ST, NW, SUITE 700, WASHINGTON, DC 20005-2210 USA SN 0006-341X J9 BIOMETRICS JI Biometrics PD JUN PY 2001 VL 57 IS 2 BP 396 EP 403 DI 10.1111/j.0006-341X.2001.00396.x PG 8 WC Biology; Mathematical & Computational Biology; Statistics & Probability SC Life Sciences & Biomedicine - Other Topics; Mathematical & Computational Biology; Mathematics GA 437QY UT WOS:000169006100011 PM 11414562 ER PT J AU Troendle, JF Frank, J AF Troendle, JF Frank, J TI Unbiased confidence intervals for the odds ratio of two independent binomial samples with application to case-control data SO BIOMETRICS LA English DT Article DE conditional; confidence interval; maximization; odds ratio; profile ID PROPORTIONS AB The problem of confidence interval construction for the odds ratio of two independent binomial samples is considered. Two methods of eliminating the nuisance parameter from the exact likelihood, conditioning and maximization, are described. A conditionally exact tail method exists by putting together upper and lower bounds. A shorter interval can be obtained by simultaneous consideration of both tails. We present here new methods that extend the tail and simultaneous approaches to the maximized likelihood. The methods are unbiased and applicable to case-control data, for which the odds ratio is important. The confidence interval procedures are compared unconditionally for small sample sizes in terms of their expected length and coverage probability. A Bayesian confidence interval method and a large-sample chi (2) procedure are included in the comparisons. C1 NICHHD, Biometry & Math Stat Branch, Div Epidemiol Stat & Prevent Res, NIH, Bethesda, MD 20892 USA. Minitab, State Coll, PA 16801 USA. RP Troendle, JF (reprint author), NICHHD, Biometry & Math Stat Branch, Div Epidemiol Stat & Prevent Res, NIH, Bldg 6100,Room 7B13, Bethesda, MD 20892 USA. NR 10 TC 9 Z9 9 U1 0 U2 1 PU INTERNATIONAL BIOMETRIC SOC PI WASHINGTON PA 1441 I ST, NW, SUITE 700, WASHINGTON, DC 20005-2210 USA SN 0006-341X J9 BIOMETRICS JI Biometrics PD JUN PY 2001 VL 57 IS 2 BP 484 EP 489 DI 10.1111/j.0006-341X.2001.00484.x PG 6 WC Biology; Mathematical & Computational Biology; Statistics & Probability SC Life Sciences & Biomedicine - Other Topics; Mathematical & Computational Biology; Mathematics GA 437QY UT WOS:000169006100022 PM 11414573 ER PT J AU Ivanova, A Berger, VW AF Ivanova, A Berger, VW TI Drawbacks to integer scoring for ordered categorical data SO BIOMETRICS LA English DT Article DE conservatism.; contingency table; linear rank test; permutation test ID TABLES AB Linear rank tests are widely used when testing for independence against stochastic order in a 2 x J contingency table with two treatments and J ordered outcome levels. For this purpose, numerical scores are assigned, possibly by default, to the J outcome levels. When the choice of scores is not apparent, integer (equally spaced) scores are often assigned. We show that this practice generally leads to unnecessarily conservative tests. The use of slightly perturbed scores will result in a less conservative and uniformly more powerful test. C1 Univ N Carolina, Dept Biostat, Chapel Hill, NC 27599 USA. NCI, DCP, Biometry Res Grp, Bethesda, MD 20892 USA. RP Ivanova, A (reprint author), Univ N Carolina, Dept Biostat, CB 7400, Chapel Hill, NC 27599 USA. NR 5 TC 14 Z9 14 U1 0 U2 0 PU INTERNATIONAL BIOMETRIC SOC PI WASHINGTON PA 1441 I ST, NW, SUITE 700, WASHINGTON, DC 20005-2210 USA SN 0006-341X J9 BIOMETRICS JI Biometrics PD JUN PY 2001 VL 57 IS 2 BP 567 EP 570 DI 10.1111/j.0006-341X.2001.00567.x PG 4 WC Biology; Mathematical & Computational Biology; Statistics & Probability SC Life Sciences & Biomedicine - Other Topics; Mathematical & Computational Biology; Mathematics GA 437QY UT WOS:000169006100034 PM 11414585 ER PT J AU Albert, PS McShane, LM Shih, JH AF Albert, PS McShane, LM Shih, JH CA US Natl Canc Inst Bladder Tumor Ma TI Latent class modeling approaches for assessing diagnostic error without a gold standard: With applications to p53 immunohistochemical assays in bladder tumors SO BIOMETRICS LA English DT Article DE biomarkers; misclassification; repeated binary data; sensitivity; specificity ID TESTS; MUTATIONS AB Improved characterization of tumors for purposes of guiding treatment decisions for cancer patients will require that accurate and reproducible assays be developed for a variety of tumor markers. No gold standards exist for most tumor marker assays. Therefore, estimates of assay sensitivity and specificity cannot he obtained unless a latent class model-based approach is used. Our goal in this article is to estimate sensitivity and specificity for p53 immunohistochemical assays of bladder tumors using data front a reproducibility study conducted by the National Cancer Institute Bladder Tumor Marker Network. We review latent class modeling approaches proposed by previous authors, and we find that many of these approaches impose assumptions about specimen heterogeneity that are not consistent with the biology of bladder tumors. We present flexible mixture model alternatives that are biologically plausible for our example, and we use them to estimate sensitivity and specificity for our p53 assay example. These mixture models are shown to offer an improvement over other methods in a variety of settings. but we caution that, in general, care must be taken in applying latent class models. C1 NCI, Biometr Res Branch, Bethesda, MD 20892 USA. NHLBI, Off Biostat Res, Bethesda, MD 20892 USA. RP Albert, PS (reprint author), NCI, Biometr Res Branch, Execut Plaza N,Room 739, Bethesda, MD 20892 USA. FU NCI NIH HHS [CA47538, CA70903, CA47526, CA47537, CA56973] NR 20 TC 46 Z9 47 U1 1 U2 10 PU INTERNATIONAL BIOMETRIC SOC PI WASHINGTON PA 1441 I ST, NW, SUITE 700, WASHINGTON, DC 20005-2210 USA SN 0006-341X J9 BIOMETRICS JI Biometrics PD JUN PY 2001 VL 57 IS 2 BP 610 EP 619 DI 10.1111/j.0006-341X.2001.00610.x PG 10 WC Biology; Mathematical & Computational Biology; Statistics & Probability SC Life Sciences & Biomedicine - Other Topics; Mathematical & Computational Biology; Mathematics GA 437QY UT WOS:000169006100040 PM 11414591 ER PT J AU Jankowsky, JL Slunt, HH Ratovitski, T Jenkins, NA Copeland, NG Borchelt, DR AF Jankowsky, JL Slunt, HH Ratovitski, T Jenkins, NA Copeland, NG Borchelt, DR TI Co-expression of multiple transgenes in mouse CNS: a comparison of strategies SO BIOMOLECULAR ENGINEERING LA English DT Review ID AMYLOID PRECURSOR PROTEIN; ALZHEIMERS-DISEASE; IN-VIVO; NUCLEAR TRANSFER; MICE; PRESENILIN-1; IDENTIFICATION; RESCUES; BRAINS; PS1 AB The introduction of two transgenes into one animal is increasingly common as transgenic experiments become more sophisticated. In this study we examine two strategies for creating double transgenic founders from a single microinjection. In the first approach, two constructs, each with its own promoter element, were coinjected into the pronucleus. In the second approach, both transgenes were cloned into one vector, separated by an internal ribosomal entry site (IRES), and placed under control of a single promoter. Both strategies save time and increase the percentage of double transgenic offspring over the standard method of mating single transgenic lines. However, despite high transgene copy numbers, the bicistronic lines did not show robust expression of either protein. Copy number and protein expression correlated much better in the coinjected lines, with expression levels in one line approaching that observed in some of our best single transgenic controls. Thus we recommend coinjection of individual plasmids for the generation of multiply transgenic founders. (C) 2001 Elsevier Science B.V. All rights reserved. C1 Johns Hopkins Sch Medt, Dept Pathol, Baltimore, MD 21205 USA. Johns Hopkins Sch Medt, Dept Neurosci, Baltimore, MD 21205 USA. NCI, Mouse Canc Genet Program, Frederick Canc Res & Dev Ctr, Frederick, MD 21702 USA. RP Borchelt, DR (reprint author), Johns Hopkins Sch Medt, Dept Pathol, 720 Rutland Ave,558 Ross Res Bldg, Baltimore, MD 21205 USA. FU NIA NIH HHS [1 P01 AG 14248] NR 26 TC 352 Z9 360 U1 5 U2 26 PU ELSEVIER SCIENCE BV PI AMSTERDAM PA PO BOX 211, 1000 AE AMSTERDAM, NETHERLANDS SN 1389-0344 J9 BIOMOL ENG JI Biomol. Eng. PD JUN PY 2001 VL 17 IS 6 BP 157 EP 165 DI 10.1016/S1389-0344(01)00067-3 PG 9 WC Biochemical Research Methods; Biotechnology & Applied Microbiology; Genetics & Heredity SC Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology; Genetics & Heredity GA 433UN UT WOS:000168778300001 PM 11337275 ER PT J AU Schroeder, BG Barry, CE AF Schroeder, BG Barry, CE TI The specificity of methyl transferases involved in trans mycolic acid biosynthesis in Mycobacterium tuberculosis and Mycobacterium smegmatis SO BIOORGANIC CHEMISTRY LA English DT Article ID CELL-WALL; ESCHERICHIA-COLI; IDENTIFICATION; SENSITIVITY; ADAPTATION; FLUIDITY; GROWTH AB Trans mycolic acid content is directly related to cell wall fluidity and permeability in mycobacteria. Carbon-13 NMR spectroscopy of mycolic acids isolated from Mycobacterium tuberculosis (MTB) and Mycobacterium smegmatis (MSM) fed C-13-labeled precursor molecules was used to probe the biosynthetic pathways that modify mycolic acids. Heteronuclear correlation spectroscopy (HMQC) of ketomycolic acid from MTB allowed assignment of the complete C-13-NMR spectrum. Incorporation patterns from [1-C-13]-acetate and [2-C-13]-acetate feeding experiments suggested that the mero chain and alpha branch of mycolic acids are both synthesized by standard fatty acid biosynthetic reactions. [C-13-methyl]-L-methionine was used to specifically label carbon atoms derived from the action of the methyl transferases involved in meromycolate modification. To enrich for trans mycolic acids a strain of MTB overexpressing the mmal gene was labeled. Carbon-carbon coupling was observed in mycolate samples doubly labeled with C-13-acetate and [C-13-methyl]-L-methionine and this information was used to assess positional specificity of methyl transfer. in MTB such methyl groups were found to occur exclusively on carbons derived from the 2 position of acetate. while in MSM they occurred only on carbons derived from the 1 position. These results suggest that the MSM methyltransferase MMAS-1 operates in an inverted manner to that of MTB. C1 NIAID, TB Res Sect, Host Def Lab, NIH, Rockville, MD 20852 USA. RP Barry, CE (reprint author), NIAID, TB Res Sect, Host Def Lab, NIH, Twinbrook II,Room 239,12441 Parklawn Dr, Rockville, MD 20852 USA. RI Barry, III, Clifton/H-3839-2012 FU Intramural NIH HHS [Z01 AI000783-11] NR 31 TC 15 Z9 15 U1 0 U2 4 PU ACADEMIC PRESS INC PI SAN DIEGO PA 525 B ST, STE 1900, SAN DIEGO, CA 92101-4495 USA SN 0045-2068 J9 BIOORG CHEM JI Bioorganic Chem. PD JUN PY 2001 VL 29 IS 3 BP 164 EP 177 DI 10.1006/bioo.2001.1207 PG 14 WC Biochemistry & Molecular Biology; Chemistry, Organic SC Biochemistry & Molecular Biology; Chemistry GA 452AX UT WOS:000169835900006 PM 11437392 ER PT J AU Burke, TR Yao, ZJ Gao, Y Wu, JX Zhu, XF Luo, JH Guo, RB Yang, DJ AF Burke, TR Yao, ZJ Gao, Y Wu, JX Zhu, XF Luo, JH Guo, RB Yang, DJ TI N-terminal carboxyl and tetrazole-containing amides as adjuvants to Grb2 SH2 domain ligand binding SO BIOORGANIC & MEDICINAL CHEMISTRY LA English DT Article ID STRUCTURE-BASED DESIGN; PHOSPHOTYROSINE-CONTAINING PEPTIDE; PHOSPHATE-CONTAINING LIGANDS; TYROSINE KINASE ZAP-70; HIGH-AFFINITY; SIGNAL-TRANSDUCTION; DRUG DESIGN; INHIBITORS; POTENT; DISCOVERY AB High affinity binding of peptides to Src homology 2 (SH2) domains, often requires the presence of phosphotyrosyl (pTyr) or pTyr-mimicking moieties in the N-terminal position of the binding ligand. Several reports have shown that N-alpha-acylation of the critical pTyr residue can result in increased SH2 domain binding potency. For Grb2 SH2 domains which recognize pTyr-Xxx-Asn-NH2 motifs, significant potency enhancement can be incurred by N-alpha-(3-amino)Z derivatization of tripeptides such as pTyr-Xxe-Asn-NH2. Using ligands based on the high affinity pY-Ac(6)c-Asn-(naphthylpropylamide) motif, (where Ac(6)c = 1-aminocyclohex-anecarboxylic acid), additional reports have shown moderate potentiating effects of N-alpha-oxalyl derivatization. The current study examined variations of the N-alpha-oxalyl theme in the context of a Xxx-Ac(6)c-Asn-(naphthylpropylamide) platform, where Xxx = the hydrolytically stable pTyr mimetics phosphonomethyl phenylalanine (Pmp) or carboxymethyl phenylalanine (Cmf). The effects of N-alpha-(3-amino)Z derivatization were also investigated for this platform, to ascertain whether the large binding enhancement reported for tripeptides such as pTyr-Ile-Asn-NH2 could be observed. In ELISA-based extracellular Grb2 SH2 domain binding assays, it was found for the Pmp-based series, that extending the oxalyl carboxyl out by one methylene unit or replacing carboxyl functionality with a tetrazole isostere, resulted in binding potency greater than the parent N-alpha-acetyl-containing compound, with enhancement approximating that observed for the N-alpha-oxalyl derivative. When Cmf was used as the pTyr mimetic, only modest differences in IC50 values were observed for the series. Examination of the N-alpha-(3-amino)Z derivatized Pmp-Ac(6)c-Asn-(naphthylpropylamide), showed that binding affinity was reduced relative to the parent N-alpha-acetyl analogue, in contrast to the reported significant enhancement of affinity observed with other peptide ligands. Treatment of MDA-453 tumor cells, which are mitogenically driven through erbB-2 tyrosine kinase-dependent pathways, with Pmp-containing inhibitors resulted in growth inhibition, with the N-alpha-oxalyl and N-alpha-malonyl-containing compounds exhibiting IC50 values (4.3 and 4.6 muM, respectively) approximately five-fold lower than the parent N-alpha-acetyl-containing compound. Tetrazole and N-alpha-(3-amino)Z-containing inhibitors were from two- to four-fold less potent than these latter analogues in the growth inhibition assays. (C) 2001 Elsevier Science Ltd. All rights reserved. C1 NCI, Med Chem Lab, Div Basic Sci, NIH, Frederick, MD 21702 USA. Georgetown Univ, Lombardi Canc Ctr, Dept Oncol, Washington, DC 20007 USA. RP Burke, TR (reprint author), NCI, Med Chem Lab, Div Basic Sci, NIH, Bldg 376,FCRDC, Frederick, MD 21702 USA. RI Burke, Terrence/N-2601-2014; Yao, Zhu-Jun/E-7635-2015 NR 53 TC 17 Z9 17 U1 0 U2 4 PU PERGAMON-ELSEVIER SCIENCE LTD PI OXFORD PA THE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLAND SN 0968-0896 J9 BIOORGAN MED CHEM JI Bioorg. Med. Chem. PD JUN PY 2001 VL 9 IS 6 BP 1439 EP 1445 DI 10.1016/S0968-0896(01)00014-1 PG 7 WC Biochemistry & Molecular Biology; Chemistry, Medicinal; Chemistry, Organic SC Biochemistry & Molecular Biology; Pharmacology & Pharmacy; Chemistry GA 444NB UT WOS:000169404600006 PM 11408162 ER PT J AU Ospeck, M Eguiluz, VM Magnasco, MO AF Ospeck, M Eguiluz, VM Magnasco, MO TI Evidence of a hoof bifurcation in frog hair cells SO BIOPHYSICAL JOURNAL LA English DT Article ID ION-DEPENDENT CONDUCTANCES; RANA-CATESBEIANA; BULL-FROG; TURTLE; NONLINEARITIES; TRANSDUCTION; RESONANCE; HEARING; STIMULI AB The membrane potential of hair cells in the low-frequency hearing organ of the bullfrog, the amphibian papilla, sinusoidally oscillates at small amplitude in the absence of acoustical input. We stimulate the cell with a series of periodic currents close to this natural frequency and observe that its current-to-voltage transfer function is compressively nonlinear, having a large gain for small stimuli and a smaller gain for larger currents. Along with the spontaneous oscillation, this implies that the cell is poised close to a dynamical instability such as a Hopf bifurcation, because distant from the instability the transfer function becomes linear. The cell's frequency selectivity is enhanced for small stimuli. Simulations show that the cell's membrane capacitance is effectively reduced due to a current gain provided by this dynamical instability. We propose that the Hopf resonance is widely used by transducer cells on the sensory periphery to achieve small-signal amplification. C1 NIDCD, Biophys Sect, Lab Cellular Biol, NIH, Bethesda, MD 20892 USA. Niels Bohr Inst, Ctr Chaos & Turbulence Studies, DK-2100 Copenhagen O, Denmark. Rockefeller Univ, Phys Math Lab, New York, NY 10021 USA. RP Ospeck, M (reprint author), NIDCD, Biophys Sect, Lab Cellular Biol, NIH, Bethesda, MD 20892 USA. RI Eguiluz, Victor/B-9655-2008 OI Eguiluz, Victor/0000-0003-1133-1289 FU NIDCD NIH HHS [DC00241] NR 28 TC 37 Z9 38 U1 0 U2 1 PU BIOPHYSICAL SOCIETY PI BETHESDA PA 9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3998 USA SN 0006-3495 J9 BIOPHYS J JI Biophys. J. PD JUN PY 2001 VL 80 IS 6 BP 2597 EP 2607 PG 11 WC Biophysics SC Biophysics GA 437QX UT WOS:000169006000010 PM 11371437 ER PT J AU Guia, A Stern, MD Lakatta, EG Josephson, IR AF Guia, A Stern, MD Lakatta, EG Josephson, IR TI Ion concentration-dependence of rat cardiac unitary L-type calcium channel conductance SO BIOPHYSICAL JOURNAL LA English DT Article ID SMOOTH-MUSCLE CELLS; PIG HEART-CELLS; CA2+ CHANNELS; VENTRICULAR MYOCYTES; CA-2+ CHANNELS; CURRENTS; VOLTAGE; STIMULATION; MODULATION; PERMEATION AB Little is known about the native properties of unitary cardiac L-type calcium currents (i(Ca)) measured with physiological calcium (Ca) ion concentration, and their role in excitation-contraction (E-C) coupling. Our goal was to chart the concentration-dependence of unitary conductance (gamma) to physiological Ca concentration and compare it to barium ion (Ba) conductance in the absence of agonists. In isolated, K-depolarized rat myocytes, i(Ca) amplitudes were measured using cell-attached patches with 2 to 70 mM Ca or 2 to 105 mM Ba in the pipette. At 0 mV, 2 mM of Ca produced 0.12 pA, and 2 mM of Ba produced 0.19 pA unitary currents. Unitary conductance was described by a Langmuir isotherm relationship with a maximum gamma (Ca) of 5.3 +/- 0.2 pS (n = 15), and gamma (Ba) of 15 +/- 1 pS (n = 27). The concentration producing half-maximal gamma, Kd((gamma)), was not different between Ca (1.7 +/- 0.3 mM) and Ba (1.9 +/- 0.4 mM), We found that quasi-physiological concentrations of Ca produced currents that were as easily resolvable as those obtained with the traditionally used higher concentrations, This study leads to future work on the molecular basis of E-C coupling with a physiological concentration of Ca ions permeating the Ca channel. C1 NIA, Cardiovasc Sci Lab, Gerontol Res Ctr, NIH, Baltimore, MD 21224 USA. RP NIA, Cardiovasc Sci Lab, Gerontol Res Ctr, NIH, 5600 Nathan Shock Dr, Baltimore, MD 21224 USA. EM JosephsonI@grc.nia.nih.gov NR 37 TC 34 Z9 37 U1 0 U2 1 PU CELL PRESS PI CAMBRIDGE PA 50 HAMPSHIRE ST, FLOOR 5, CAMBRIDGE, MA 02139 USA SN 0006-3495 EI 1542-0086 J9 BIOPHYS J JI Biophys. J. PD JUN PY 2001 VL 80 IS 6 BP 2742 EP 2750 PG 9 WC Biophysics SC Biophysics GA 437QX UT WOS:000169006000022 PM 11371449 ER PT J AU Karp, JE Lancet, JE Kaufmann, SH End, DW Wright, JJ Bol, K Horak, I Tidwell, ML Liesveld, J Kottke, TJ Ange, D Buddharaju, L Gojo, I Highsmith, WE Belly, RT Hohl, RJ Rybak, ME Thibault, A Rosenblatt, J AF Karp, JE Lancet, JE Kaufmann, SH End, DW Wright, JJ Bol, K Horak, I Tidwell, ML Liesveld, J Kottke, TJ Ange, D Buddharaju, L Gojo, I Highsmith, WE Belly, RT Hohl, RJ Rybak, ME Thibault, A Rosenblatt, J TI Clinical and biologic activity of the farnesyltransferase inhibitor R115777 in adults with refractory and relapsed acute leukemias: a phase 1 clinical-laboratory correlative trial SO BLOOD LA English DT Article ID ACUTE MYELOID-LEUKEMIA; ACUTE LYMPHOBLASTIC-LEUKEMIA; SIGNAL-TRANSDUCTION; RAS MUTATIONS; LAMIN-A; PROTEIN; CANCER; GENE; DAUNORUBICIN; MECHANISMS AB R115777 is a nonpeptidomimetic enzyme-specific inhibitor of farnesyl protein transferase (FT) that was developed as a potential inhibitor of Ras protein signaling, with antitumor activity in preclinical models, This study was a phase 1 trial of orally administered R115777 in 35 adults with poor-risk acute leukemias. Cohorts of patients received R115777 at doses ranging from 100 mg twice daily (bid) to 1200 mg bid for up to 21 days, Dose-limiting toxicity occurred at 1200 mg bid, with central neurotoxicity evidenced by ataxia, confusion, and dysarthria, Non-dose-limiting toxicities included reversible nausea, renal insufficiency, polydipsia, paresthesias, and myelosuppression, R115777 inhibited FT activity at 300 mg bid and farnesylation of FT substrates lamin A and HDJ-2 at 600 mg bid, Extracellular signal-regulated kinase (ERK), an effector enzyme of Ras-mediated signaling, was detected in its phosphorylated (activated) form in 8 (36.4%) of 22 pretreatment marrows and became undetectable in 4 of those 8 after one cycle of treatment. Pharmacokinetics revealed a linear relationship between dose and maximum plasma concentration or area under the curve over 12 hours at all dose levels. Weekly marrow samples demonstrated that R115777 accumulated in bone marrow in a dose-dependent fashion, with large increases in marrow drug levels beginning at 600 mg bid and with sustained levels throughout drug administration, Clinical responses occurred in 10 (29%) of the 34 evaluable patients, including 2 complete remissions, Genomic analyses failed to detect N-ras gene mutations in any of the 35 leukemias, The results of this first clinical trial of a signal transduction inhibitor in patients with acute leukemias suggest that inhibitors of FT may have important clinical antileukemic activity. (Blood, 2001;97:3361-3369) (C) 2001 by The American Society of Hematology. C1 Univ Maryland, Greenbaum Canc Ctr, Baltimore, MD 21201 USA. Univ Rochester, Ctr Canc, Rochester, NY USA. Mayo Clin, Rochester, MN USA. Janssen Res Fdn, B-2340 Beerse, Belgium. Janssen Res Fdn, Spring House, PA 19477 USA. NCI, Bethesda, MD 20892 USA. Ortho Clin Diagnost, Rochester, NY USA. Univ Iowa, Iowa City, IA USA. RP Karp, JE (reprint author), Univ Maryland, Greenbaum Canc Ctr, 22 S Greene St,Rm S9D07, Baltimore, MD 21201 USA. RI Highsmith, William/B-6175-2008 FU NCI NIH HHS [U01 CA69854] NR 55 TC 358 Z9 371 U1 0 U2 3 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1900 M STREET. NW SUITE 200, WASHINGTON, DC 20036 USA SN 0006-4971 J9 BLOOD JI Blood PD JUN 1 PY 2001 VL 97 IS 11 BP 3361 EP 3369 DI 10.1182/blood.V97.11.3361 PG 9 WC Hematology SC Hematology GA 436GJ UT WOS:000168927900008 PM 11369625 ER PT J AU Braun, MC Wang, JM Lahey, E Rabin, RL Kelsall, BL AF Braun, MC Wang, JM Lahey, E Rabin, RL Kelsall, BL TI Activation of the formyl peptide receptor by the HIV-derived peptide T-20 suppresses interleukin-12 p70 production by human monocytes SO BLOOD LA English DT Article ID HUMAN-IMMUNODEFICIENCY-VIRUS; PROTEIN-COUPLED RECEPTOR; BLOOD MONONUCLEAR-CELLS; ZIPPER-LIKE DOMAIN; CD4(+) T-CELLS; INFECTED INDIVIDUALS; DENDRITIC CELLS; IMMUNE FUNCTION; TYPE-1 GP41; IN-VITRO AB It has been proposed that in the early stages of human immunodeficiency (HIV) infection, before the loss of CD4(+) T cells, inhibition of IL-12 production from host antigen-presenting cells plays a critical role in the suppression of T-helper cell type 1 responses. Activation of the G(i)-protein-coupled high-affinity N-formyl peptide receptor by f-met-leu-phe and HIV-derived peptide T-20-suppressed IL-12 p70 production from human monocytes in response to both T-cell-dependent and T-cell-independent stimulation are reported. Activation of the low-affinity N-formyl peptide receptor by the HIV-derived F-peptide suppressed IL-12 production more modestly. This suppression was pertussis toxin sensitive and was selective for IL-12; the production of IL-10, transforming growth factor-p, and tumor necrosis factor-alpha was unaltered. The production of IL-12 p70 by dendritic cells was unaffected by these peptides despite functional expression of the high-affinity fMLP receptor. These findings provide a potential direct mechanism for HIV-mediated suppression of IL-12 production and suggest a broader role for G-protein-coupled receptors in the regulation of Innate immune responses. (Blood, 2001;97:3531-3536) (C) 2001 by The American Society of Hematology. C1 NIAID, Mucosal Immun Sect, Immune Cell Interact Unit, NIH, Bethesda, MD 20892 USA. NIAID, Clin Invest Lab, Cytokine Biol Unit, NIH, Bethesda, MD 20892 USA. NCI, Frederick Canc Res & Dev Ctr, Div Basic Sci, Mol Immunoregulat Lab, Frederick, MD USA. RP Kelsall, BL (reprint author), NIAID, Mucosal Immun Sect, Immune Cell Interact Unit, NIH, Bldg 10,Rm 11N238,10 Ctr Dr, Bethesda, MD 20892 USA. NR 54 TC 20 Z9 20 U1 1 U2 1 PU AMER SOC HEMATOLOGY PI WASHINGTON PA 1900 M STREET. NW SUITE 200, WASHINGTON, DC 20036 USA SN 0006-4971 J9 BLOOD JI Blood PD JUN 1 PY 2001 VL 97 IS 11 BP 3531 EP 3536 DI 10.1182/blood.V97.11.3531 PG 6 WC Hematology SC Hematology GA 436GJ UT WOS:000168927900030 PM 11369647 ER PT J AU Maguire, EA Vargha-Khadem, F Mishkin, M AF Maguire, EA Vargha-Khadem, F Mishkin, M TI The effects of bilateral hippocampal damage on fMRI regional activations and interactions during memory retrieval SO BRAIN LA English DT Article DE autobiographical memory; effective connectivity; fMRI; hippocampus; hypoxia ID POSITRON-EMISSION-TOMOGRAPHY; PREFRONTAL CORTEX; EPISODIC MEMORY; RECOGNITION MEMORY; AUTOBIOGRAPHICAL MEMORY; RETROSPLENIAL CORTEX; RETROGRADE-AMNESIA; ALZHEIMERS-DISEASE; DECLARATIVE MEMORY; PICKS-DISEASE AB Using functional magnetic resonance imaging (fMRI) we examined successful retrieval of real-world memories in a patient (Jon) with selective bilateral hippocampal pathology resulting from perinatal hypoxia compared with healthy control subjects. Jon activated the same brain regions during memory retrieval as control subjects, both medial and lateral on the left. In contrast to controls, Jon also activated many homologous regions on the right. In spite of having 50% volume loss bilaterally in his hippocampi, retrieval in Jon was associated with increased activation of the hippocampi. Furthermore, hippocampal activity, as with the controls, was differential, being most responsive to retrieval of autobiographical events compared,vith other memory types (autobiographical facts, public events, general knowledge). Jon made a distinction between events that the control subjects did not make, namely that some of the autobiographical and public events he clearly remembered, while others he found that he knew about but did not truly remember. His hippocampi and medial frontal cortex were significantly more active during retrieval of events for which he had clear and conscious recollection compared with those he knew as much about, including the context, but could not remember experiencing. Although Jon activates the same network of brain regions as the controls (albeit bilaterally), and with the same pattern of response in the hippocampus, the communication between regions differs from controls with regard to hippocampal-cortical connectivity. In controls there was increased effective connectivity between parahippocampal cortex and hippocampus, specifically during the retrieval of autobiographical events. In contrast, this increase was not apparent in Jon; rather, retrieval of autobiographical events elicited greater interaction between the hippocampus and retrosplenial cortex, and also increased interaction between retrosplenial and medial frontal cortex. This study underlines the value of scanning patients using fMRI while they undertake tasks they can perform, in this case allowing us to confirm the functionality of remaining tissue in the damaged hippocampi, and to appreciate the neural basis of a distinction (remember/know) that control subjects do not make. Besides refining our knowledge of the hippocampal role in autobiographical event memory, this study indicates that recruitment of bilateral regions during memory retrieval, and altered patterns of effective connectivity between brain regions may be important indicators of disordered memory. C1 UCL, Inst Neurol, Wellcome Dept Cognit Neurol, London WC1N 3BG, England. UCL, Dev Cognit Neurosci Unit, Inst Child Hlth, London WC1E 6BT, England. NIMH, Neuropsychol Lab, Bethesda, MD 20892 USA. RP Maguire, EA (reprint author), UCL, Inst Neurol, Wellcome Dept Cognit Neurol, 12 Queen Sq, London WC1N 3BG, England. EM e.maguire@fil.ion.ucl.ac.uk RI Vargha-Khadem, Faraneh/C-2558-2008 NR 62 TC 177 Z9 179 U1 4 U2 34 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0006-8950 J9 BRAIN JI Brain PD JUN PY 2001 VL 124 BP 1156 EP 1170 DI 10.1093/brain/124.6.1156 PN 6 PG 15 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 440EL UT WOS:000169162900011 PM 11353732 ER PT J AU Ziemann, U Muellbacher, W Hallett, M Cohen, LG AF Ziemann, U Muellbacher, W Hallett, M Cohen, LG TI Modulation of practice-dependent plasticity in human motor cortex SO BRAIN LA English DT Article DE practice-dependent plasticity; GABA-related inhibition; modulation of plasticity; human motor cortex; transcranial magnetic stimulation ID LONG-TERM POTENTIATION; TRANSCRANIAL MAGNETIC STIMULATION; ISCHEMIC NERVE BLOCK; HORIZONTAL CONNECTIONS; CORTICAL EXCITABILITY; SENSORY CORTEX; TRANSIENT DEAFFERENTATION; INTRACORTICAL INHIBITION; MOVEMENT REPRESENTATIONS; DYNAMIC ORGANIZATION AB Motor practice may lead to expansion of trained representations in the motor cortex, but it is unknown whether this practice-dependent plasticity can be purposefully enhanced or depressed. Evidence, mainly based on animal experiments, indicates that the activity of GABA-related cortical inhibition is important in controlling the extent to which plasticity may occur. We tested the role of GABA in modulating practice-dependent plasticity in the human motor cortex. A decrease in GABA-related cortical inhibition was achieved by ischaemic nerve block (INB) in the hand by deafferentation/deefferentation and an increase was achieved by administration of the GABAA receptor agonist lorazepam, In Experiment 1, healthy subjects performed motor practice (MP), consisting of repeated ballistic contractions of the biceps muscle in the absence (MP alone) or presence of INB (MP+INB), Changes in the biceps motor cortex representation were assessed by transcranial magnetic stimulation (TMS), MP+INB resulted in a dramatic increase in the size of the motor evoked potential (MEP) and in paired-pulse excitability compared with mild or no changes in the MP-alone and INB-alone conditions. In Experiment 2, this dramatic increase in biceps representation induced by MP+INB was replicated when subjects were pretreated with placebo, but this increase was prevented or even switched to a decrease when subjects were pretreated with lorazepam, These findings indicate that a decrease in GABA-related inhibition facilitates practice-dependent plasticity in the human motor cortex, whereas an increase depresses it. In Experiment 3, practice-dependent plasticity (assessed by TMS, as in the first two experiments) was also tested at the behavioural level, The dramatic increase in biceps MEP size induced by MP+INB was paralleled by an increase in peak acceleration of the fastest elbow flexion movements, Similarly, the lack of change in MEP size in the MP-alone condition was paralleled by a lack of change in peak acceleration. We propose that changes in GABA activity may be instrumented to modulate plasticity purposefully; for instance, to enhance plastic change and recovery of function after a lesion in neurological patients. C1 Univ Frankfurt, Neurol Clin, D-60590 Frankfurt, Germany. Natl Inst Neurol Disorders & Stroke, Human Cort Physiol Sect, NIH, Bethesda, MD USA. Natl Inst Neurol Disorders & Stroke, Human Motor Control Sect, NIH, Bethesda, MD USA. RP Ziemann, U (reprint author), Univ Frankfurt, Neurol Clin, Schleusenweg 2-16, D-60590 Frankfurt, Germany. NR 84 TC 261 Z9 266 U1 0 U2 17 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0006-8950 J9 BRAIN JI Brain PD JUN PY 2001 VL 124 BP 1171 EP 1181 DI 10.1093/brain/124.6.1171 PN 6 PG 11 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 440EL UT WOS:000169162900012 PM 11353733 ER PT J AU Winterer, G Mulert, C Mientus, S Gallinat, J Schlattmann, P Dorn, H Herrmann, WM AF Winterer, G Mulert, C Mientus, S Gallinat, J Schlattmann, P Dorn, H Herrmann, WM TI P300 and LORETA: Comparison of normal subjects and schizophrenic patients SO BRAIN TOPOGRAPHY LA English DT Article DE P300; LORETA; schizophrenia; left hemisphere; cortical network ID EVENT-RELATED POTENTIALS; RESOLUTION ELECTROMAGNETIC TOMOGRAPHY; SUPERIOR TEMPORAL GYRUS; TO-NOISE RATIO; INTRACEREBRAL POTENTIALS; AUDITORY ODDBALL; VISUAL-STIMULI; RARE TARGET; OSCILLATORY RESPONSES; ENDOGENOUS POTENTIALS AB It was the aim of the present study 1) to investigate how many cortical activity maxima of scalp-recorded P300 are detected by Low Resolution Electromagentic Tomography (LORETA) when analyses are performed with high time-resolution, 2) to see if the resulting LORETA-solution is in accordance with intracortical recordings as reported by others and 3) to compare the given pattern of cortical activation maxima in the P300-timeframe between schizophrenic patients and normal controls. Current density analysis was performed in 3-D Talairach space with high time resolution i.e. in 6 ms steps. This was done during an auditory choice reaction paradigm separately for normal subjects and schizophrenic patients with subsequent group comparisons. In normal subjects, a sequence of at least seven cortical activation maxima was found between 240-420ms poststimulus: the prefrontal cortex, anterior or medial cingulum, posterior cingulum, parietal cortex, temporal lobe, prefrontal cortex, medial or anterior cingulum. Within the given limits of spatial resolution, this sequential maxima distribution largely met the expectations from reports on intracranial recordings and functional neuroimaging studies. However, localization accuracy was higher near the central midline than at lateral aspects of the brain. Schizophrenic patients less activated their cortex in a widespread area mainly in the left hemisphere including the prefrontal cortex, posterior cingulum and the temporal lobe. From these analyses and comparsions, with intracranial recordings as reported by others, it is concluded that LORETA correctly localizes P300-related cortical activity maxima on the basis of 19 electrodes except for lateral cortical aspects which is most likely an edge-phenomenon. The data further suggest that the P300-deficit in schizophrenics involves an extended cortical network of the left hemisphere at several steps in time during the information processing stream. C1 NIMH, Mol Neuroimaging Unit, Clin Brain Disorders Branch, NIH, Bethesda, MD 20892 USA. Free Univ Berlin, Dept Psychiat, Lab Clin Psychophysiol, D-1000 Berlin, Germany. RP Winterer, G (reprint author), NIMH, Mol Neuroimaging Unit, Clin Brain Disorders Branch, NIH, Bldg 10,Room 4S229A,MSC 1379, Bethesda, MD 20892 USA. RI Schlattmann, Peter/B-5289-2008; Mulert, Christoph/F-2576-2012 NR 87 TC 89 Z9 91 U1 0 U2 2 PU KLUWER ACADEMIC-HUMAN SCIENCES PRESS PI NEW YORK PA 233 SPRING ST, NEW YORK, NY 10013-1578 USA SN 0896-0267 J9 BRAIN TOPOGR JI Brain Topogr. PD SUM PY 2001 VL 13 IS 4 BP 299 EP 313 DI 10.1023/A:1011184814194 PG 15 WC Clinical Neurology; Neurosciences SC Neurosciences & Neurology GA 464PR UT WOS:000170541400006 PM 11545159 ER PT J AU Zheng, T Holford, TR Mayne, ST Owens, PH Zhang, Y Zhang, B Boyle, P Zahm, SH AF Zheng, T Holford, TR Mayne, ST Owens, PH Zhang, Y Zhang, B Boyle, P Zahm, SH TI Lactation and breast cancer risk: a case-control study in Connecticut SO BRITISH JOURNAL OF CANCER LA English DT Article DE breast cancer; breastfeeding; lactation; case-control study ID MENOPAUSAL STATUS; CHINESE-WOMEN; HISTORY; ASSOCIATION AB In this report, we examined the relationship between lactation and breast cancer risk, in a case-control study of breast cancer, conducted in Connecticut between 1994 and 1998. Included were 608 incident breast cancer cases and 609 age frequency matched controls, aged 30-80 years old, Cases and controls were interviewed by trained study interviewers, using a standardized, structured questionnaire, to obtain information on lactation and other major risk factors, Parous women who reported ever lactation had a borderline significantly reduced risk of breast cancer (OR = 0.83, 95% CI, 0.63-1.09). An OR of 0.53 (95% CI, 0.27-1.04) was observed in those having breastfed more than 3 children compared to those who never lactated. Women having breastfed their first child for more than 13 months had an OR of 0.47 (95% CI, 0.23-0.94) compared to those who never breastfed. Lifetime duration of lactation also showed a risk reduction while none of the ORs were statistically significant. Further stratification by menopausal status showed a risk reduction related to lactation for both pre- and postmenopausal women, while the relationship is less consistent for the latter. These results support an inverse association between breastfeeding and breast cancer risk. (C) 2001 Cancer Research Campaign. C1 Yale Univ, Sch Med, Dept Epidemiol & Publ Hlth, New Haven, CT 06520 USA. McGill Univ, Dept Epidemiol & Biostat, Montreal, PQ H3A 1A2, Canada. European Inst Oncol, Dept Epidemiol & Biostat, I-20141 Milan, Italy. NCI, Div Epidemiol & Genet, Bethesda, MD 20892 USA. RP Zheng, T (reprint author), Yale Univ, Sch Med, Dept Epidemiol & Publ Hlth, 60 Coll, New Haven, CT 06520 USA. RI Boyle, Peter/A-4380-2014; Zahm, Shelia/B-5025-2015 OI Boyle, Peter/0000-0001-6251-0610; FU NCI NIH HHS [CA-62986] NR 23 TC 42 Z9 45 U1 0 U2 3 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 JUN 1 PY 2001 VL 84 IS 11 BP 1472 EP 1476 DI 10.1054/bjoc.2001.1793 PG 5 WC Oncology SC Oncology GA 444RZ UT WOS:000169414900009 PM 11384096 ER PT J AU Foster, CB Zhu, SX Erichsen, HC Lehrnbecher, T Hart, ES Choi, E Stein, S Smith, MW Steinberg, SM Imbach, P Kuhne, T Chanock, SJ AF Foster, CB Zhu, SX Erichsen, HC Lehrnbecher, T Hart, ES Choi, E Stein, S Smith, MW Steinberg, SM Imbach, P Kuhne, T Chanock, SJ CA Early Chronic ITP Study Grp TI Polymorphisms in inflammatory cytokines and Fc gamma receptors in childhood chronic immune thrombocytopenic purpura: a pilot study SO BRITISH JOURNAL OF HAEMATOLOGY LA English DT Article DE cytokines; Fc gamma receptors; chronic immune thrombocytopenic purpura; genetic associations ID VARIANT GENOTYPES; PHAGOCYTOSIS; PROMOTER; SARCOMA; BINDING; RIIIA; MEN AB Inflammatory cytokines and low-affinity Fc gamma receptor (Fc gammaR) polymorphisms were investigated in 37 children with chronic immune thrombocytopenic purpura (cITP) and 218 controls. Genotype analysis included common variants in the regulatory regions of cytokines, TNF, LTA, IL1RN, IL1A, IL1B, IL4, IL6 and IL10, and structural variants of the low affinity Fc gamma Rs, FCGR2A, FCGR3A and FCGR3B. Associations were observed for TNF (P = 0.0032), LTA (P = 0.019), FCGR3A (P = 0.038) and FCGR3B (P = 0.0034). Two combinations of genotypes (TNF and FCGR3A; P = 0.0003, and LTA and FCGR3B; P = 0.011) were significantly associated with cITP, These results provide preliminary evidence that variant genotypes of Fc gamma Rs and cytokines contribute to cITP pathogenesis. C1 NCI, Ctr Adv Technol, Pediat Oncol Branch, Immunocompromised Host Sect,NIH, Gaithersburg, MD 20877 USA. NCI, Sci Applicat Int Corp, Frederick, MD 21701 USA. NCI, Biostat & Data Management Sect, NIH, Bethesda, MD 20892 USA. Univ Basel, Basel, Switzerland. RP Chanock, SJ (reprint author), NCI, Ctr Adv Technol, Pediat Oncol Branch, Immunocompromised Host Sect,NIH, 8717 Grovemont Circle, Gaithersburg, MD 20877 USA. RI Smith, Michael/B-5341-2012; Choi, Eun Hwa/J-5691-2012 NR 16 TC 56 Z9 59 U1 0 U2 1 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 JUN PY 2001 VL 113 IS 3 BP 596 EP 599 DI 10.1046/j.1365-2141.2001.02807.x PG 4 WC Hematology SC Hematology GA 444YR UT WOS:000169428600006 PM 11380443 ER PT J AU Kupka, RW Nolen, WA Altshuler, LL Denicoff, KD Frye, MA Leverich, GS Keck, PE McElroy, SL Rush, AJ Suppes, T Post, RM AF Kupka, RW Nolen, WA Altshuler, LL Denicoff, KD Frye, MA Leverich, GS Keck, PE McElroy, SL Rush, AJ Suppes, T Post, RM TI The Stanley Foundation Bipolar Network 2. Preliminary summary of demographics, course of illness and response to novel treatments SO BRITISH JOURNAL OF PSYCHIATRY LA English DT Article; Proceedings Paper CT 1st European Stanley-Foundation Symposium on Bipolar Disorder CY SEP 24-25, 1998 CL ROYAL SOC, LONDON, ENGLAND SP Stanley Fdn HO ROYAL SOC ID AFFECTIVE-DISORDER; SUBSTANCE-ABUSE; MOOD DISORDERS; LAMOTRIGINE; SCALE; HYPOTHYROIDISM; EPIDEMIOLOGY; ASSOCIATION; MONOTHERAPY; RELIABILITY AB Background The Stanley Foundation Bipolar Network (SFBN) evaluates treatments, course and clinical and neurobiological markers of response in bipolar illness. Aims To give a preliminary summary of emerging findings in these areas. Method Studies with established and potentially antimanic, antidepressant and mood-stabilising agents range from open case series to double-blind randomised clinical trials, and use the same core assessment methodology, thereby optimising the comparability of the outcomes. The National Institute of Mental Health Life Chart Method is the core instrument for retrospective and prospective longitudinal illness description. Results The first groups of patients enrolled show a considerable degree of past and present symptomatology, psychiatric comorbidity and functional impairment. There are associations of both genetic and early environmental factors with more severe courses of illness. Open case series with add-on olanzapine, lamotrigine, gabapentin or topiramate show a differential spectrum of effectiveness in refractory patients. Conclusions The SFBN provides important new data for the understanding and treatment of bipolar disorder. C1 Altrecht Ctr Mental Hlth Care, NL-3512 PZ Utrecht, Netherlands. Univ Utrecht, Med Ctr, Utrecht, Netherlands. Univ Calif Los Angeles, Los Angeles, CA 90024 USA. VA Med Ctr, Los Angeles, CA USA. NIMH, Biol Psychiat Branch, Bethesda, MD 20892 USA. Univ Cincinnati, Coll Med, Biol Psychiat Program, Cincinnati, OH 45221 USA. Univ Texas, SW Med Ctr, Dallas, TX USA. RP Altrecht Ctr Mental Hlth Care, Vrouwjuttenhof 18, NL-3512 PZ Utrecht, Netherlands. EM kupka@hcrg.nl RI Nolen, Willem/E-9006-2014 NR 43 TC 23 Z9 24 U1 3 U2 4 PU ROYAL COLLEGE OF PSYCHIATRISTS PI LONDON PA BRITISH JOURNAL OF PSYCHIATRY 17 BELGRAVE SQUARE, LONDON SW1X 8PG, ENGLAND SN 0007-1250 EI 1472-1465 J9 BRIT J PSYCHIAT JI Br. J. Psychiatry PD JUN PY 2001 VL 178 SU 41 BP S177 EP S183 DI 10.1192/bjp.178.41.s177 PG 7 WC Psychiatry SC Psychiatry GA 443QD UT WOS:000169351100012 PM 11450180 ER PT J AU Manji, HK Moore, GJ Chen, G AF Manji, HK Moore, GJ Chen, G TI Bipolar disorder: leads from the molecular and cellular mechanisms of action of mood stabilisers SO BRITISH JOURNAL OF PSYCHIATRY LA English DT Article; Proceedings Paper CT 1st European Stanley Foundation Symposium on Bipolar Disorder CY SEP 24-25, 1998 CL ROYAL SOC, LONDON, ENGLAND SP Stanley Fdn HO ROYAL SOC ID PROTEIN-KINASE-C; SIGNAL-TRANSDUCTION PATHWAYS; GLYCOGEN-SYNTHASE KINASE-3; MANIC-DEPRESSIVE ILLNESS; DNA-BINDING ACTIVITY; GENOME-WIDE SEARCH; 2ND MESSENGER SYSTEMS; OLD ORDER AMISH; GENE-EXPRESSION; LITHIUM TREATMENT AB Background New research is dramatically altering our understanding of the molecular mechanisms underlying neuronal communication. Aim To elucidate the molecular mechanisms underlying the therapeutic effects of mood stabilisers. Method Results from integrated clinical and laboratory studies are reviewed. Results Chronic administration of lithium and valproate produced a striking reduction in protein kinase C (PKC) isozymes in rat frontal cortex and hippocampus. In a small study, tamoxifen (also a PKC inhibitor) had marked antimanic efficacy. Both lithium and valproate regulate the DNA binding activity of the activator protein I family of transcription factors, Using m RNA differential display. it was also shown that chronic administration of lithium and valproate modulates expression of several genes. An exciting finding is that of a robust elevation in the levels of the cytoprotective protein, bcl-2. Conclusions The results suggest that regulation of signalling pathways may play a major part in the long-term actions of mood stabilisers. Additionally mood stabilisers may exert underappreciated neuroprotective effects. C1 Wayne State Univ, Sch Med, Mol Pathophysiol Lab, Detroit, MI 48201 USA. RP Manji, HK (reprint author), NIMH, Mol Pathophysiol Lab, Bld 49,Room B1EE16,49 Convent Dr,MSC 4405, Bethesda, MD 20892 USA. RI Moore, Gregory/E-7184-2010; Chen, Guang/A-2570-2017 OI Moore, Gregory/0000-0001-8541-3194; NR 141 TC 40 Z9 41 U1 1 U2 5 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 JUN PY 2001 VL 178 SU 41 BP S107 EP S119 DI 10.1192/bjp.178.41.s107 PG 13 WC Psychiatry SC Psychiatry GA 443QD UT WOS:000169351100002 PM 11450170 ER PT J AU Post, RM Nolen, WA Kupka, RW Denicoff, KD Leverich, GS Keck, PE McElroy, SL Rush, AJ Suppes, T Altshuler, LL Frye, MA Grunze, H Walden, J AF Post, RM Nolen, WA Kupka, RW Denicoff, KD Leverich, GS Keck, PE McElroy, SL Rush, AJ Suppes, T Altshuler, LL Frye, MA Grunze, H Walden, J TI The Stanley Foundation Bipolar Network I. Rationale and methods SO BRITISH JOURNAL OF PSYCHIATRY LA English DT Article; Proceedings Paper CT 1st European Stanley-Foundation Symposium on Bipolar Disorder CY SEP 24-25, 1998 CL ROYAL SOC, LONDON, ENGLAND SP Stanley Fdn HO ROYAL SOC ID DEPRESSIVE SYMPTOMATOLOGY IDS; REFRACTORY MOOD DISORDERS; INTRACELLULAR CALCIUM; NATIONAL-INSTITUTE; CONTROLLED TRIALS; WORKSHOP REPORT; DOUBLE-BLIND; ILLNESS; LITHIUM; PLACEBO AB Background The Stanley Foundation Bipolar Network (SFBN) was created to address the paucity of help studies in bipolar illness. Aims To describe the rationale and methods of the SFBN. Method The SFBN includes five core sites and a number of affiliated sites that have adopted consistent methodology for continuous longitudinal monitoring of patients. Open and controlled studies are performed as patients symptomatology dictates. Results The reliability of SFBN raters and the validity of the rating instruments have been established. More than 500 patients are in continuous daily longitudinal follow-up. More than 125 have been randomised to one of three of the newer antidepressants (bupropion, sertraline and venlafaxine) as adjuncts in a study of mood stabilisers and 93 to omega-3 fatty acids. A number of open clinical case series have been published. Conclusions Well-characterised patients are followed in a detailed continuous longitudinal fashion in both opportunistic case series and double-blind, randomised controlled trials with reliable and validated measures. C1 NIMH, Biol Psychiat Branch, Bethesda, MD 20892 USA. HC Rumke Grp, Utrecht, Netherlands. Univ Utrecht, Med Ctr, Utrecht, Netherlands. Stanley Ctr, Cincinnati, OH USA. Univ Texas, SW Med Ctr, Stanley Ctr, Dallas, TX USA. Univ Calif Los Angeles, Stanley Ctr, Los Angeles, CA USA. Univ Munich, Dept Psychiat, D-80539 Munich, Germany. Univ Freiburg, Dept Psychiat & Psychotherapy, Freiburg, Germany. RP NIMH, Biol Psychiat Branch, Bldg 10,Room 35239,10 Ctr Dr MSC 1272, Bethesda, MD 20892 USA. RI Nolen, Willem/E-9006-2014 NR 47 TC 66 Z9 66 U1 2 U2 4 PU ROYAL COLLEGE OF PSYCHIATRISTS PI LONDON PA BRITISH JOURNAL OF PSYCHIATRY 17 BELGRAVE SQUARE, LONDON SW1X 8PG, ENGLAND SN 0007-1250 EI 1472-1465 J9 BRIT J PSYCHIAT JI Br. J. Psychiatry PD JUN PY 2001 VL 178 SU 41 BP S169 EP S176 DI 10.1192/bjp.178.41.s169 PG 8 WC Psychiatry SC Psychiatry GA 443QD UT WOS:000169351100011 PM 11450179 ER PT J AU MacKay, C AF MacKay, C TI Reconsidering the pseudo-patient study - Commentary SO CAMBRIDGE QUARTERLY OF HEALTHCARE ETHICS LA English DT Editorial Material C1 NIH, Bethesda, MD 20892 USA. RP MacKay, C (reprint author), NIH, Bldg 10, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU CAMBRIDGE UNIV PRESS PI PORT CHESTER PA 110 MIDLAND AVE, PORT CHESTER, NY 10573-9863 USA SN 0963-1801 J9 CAMB Q HEALTHC ETHIC JI Camb. Q. Healthc. Ethics PD SUM PY 2001 VL 10 IS 3 BP 332 EP 334 DI 10.1017/S0963180101223123 PG 3 WC Health Care Sciences & Services; Health Policy & Services; Social Sciences, Biomedical SC Health Care Sciences & Services; Biomedical Social Sciences GA 439CK UT WOS:000169093100012 ER PT J AU Winn, DM Diehl, SR Brown, LM Harty, LC Bravo-Otero, E Fraumeni, JF Kleinman, DV Hayes, RB AF Winn, DM Diehl, SR Brown, LM Harty, LC Bravo-Otero, E Fraumeni, JF Kleinman, DV Hayes, RB TI Mouthwash in the etiology of oral cancer in Puerto Rico SO CANCER CAUSES & CONTROL LA English DT Article DE alcohol drinking; epidemiology; human; mouth neoplasms; mouthwashes; pharyngeal neoplasms; smoking ID AERODIGESTIVE TRACT CANCER; SQUAMOUS-CELL CANCER; PHARYNGEAL CANCER; ALCOHOL-CONSUMPTION; DENTAL FACTORS; RISK; TOBACCO; CAVITY; SMOKING; PAST AB Objectives: To determine if the risk of cancers of the mouth and pharynx is associated with mouthwash use in Puerto Rico, an area of relatively high risk. Methods: Interviews were conducted with 342 cases of oral and pharyngeal cancer registered in Puerto Rico and diagnosed between 1992 and 1995 and with 521 population-based controls regarding mouthwash use and other factors. Mouthwash-related risks were estimated using unconditional logistic regression controlling for potential confounders. Results: The adjusted odds ratio associated with using mouthwash with an alcohol content of 25% or greater was 1.0. Risks were not higher with greater frequency, years of use, or lifetime mouthwash exposure. Among tobacco and alcohol abstainers the odds ratio associated with mouthwash use was 2.8 (CI = 0.8-9.9), in contrast to 0.8 (CI = 0.4-1.7) and 0.9 (CI = 0.6-1.3) among those with light and heavy cigarette smoking/alcohol drinking behaviors, respectively. Conclusions: There was no overall increased risk of oral cancer associated with mouthwash use. An elevated, but not statistically significant, risk was observed among the small number of subjects who neither smoked cigarettes nor drank alcohol, among whom an effect of alcohol-containing mouthwash would be most likely evident. Our findings indicate the need to clarify the mechanisms of oral carcinogenesis, including the possible role of alcohol-containing mouthwash. C1 Div Canc Control & Populat Sci, Bethesda, MD 20892 USA. Natl Inst Dent & Craniofacial Res, Div Intramural Res, Bethesda, MD 20892 USA. Natl Inst Dent & Craniofacial Res, Off Director, Bethesda, MD 20892 USA. NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Univ Puerto Rico, San Juan, PR 00936 USA. RP Winn, DM (reprint author), Div Canc Control & Populat Sci, 6130 Execut Blvd,Room 5114,MSC 7395, Bethesda, MD 20892 USA. NR 53 TC 24 Z9 26 U1 0 U2 3 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 JUN PY 2001 VL 12 IS 5 BP 419 EP 429 DI 10.1023/A:1011291807468 PG 11 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 455XA UT WOS:000170052000005 PM 11545457 ER PT J AU Coates, RJ Uhler, RJ Brogan, DJ Gammon, MD Malone, KE Swanson, CA Flagg, EW Brinton, LA AF Coates, RJ Uhler, RJ Brogan, DJ Gammon, MD Malone, KE Swanson, CA Flagg, EW Brinton, LA TI Patterns and predictors of the breast cancer detection methods in women under 45 years of age (United States) SO CANCER CAUSES & CONTROL LA English DT Article DE breast cancer; detection; mammography; screening; young women ID MAMMOGRAPHY SCREENING GUIDELINES; SOCIETY GUIDELINES; SELF-EXAMINATION; RISK-FACTORS; CARCINOMA; SURVIVAL; HISTORY; POPULATION; ADHERENCE; PATIENT AB Objectives: Few studies have examined methods by which breast cancers are detected, and only one study has been published on predictors of those methods. This study examined patterns and predictors of breast cancer detection methods during 1990-1992 among women age 20-44. Methods: In-person interview and medical record data were obtained during a population-based case-control study of 1619 women newly diagnosed with breast cancer in three areas of the United States (US). Results: Seventy-one percent of the cancers were identified by self-detection, 9% by routine clinical breast exam (CBE), and 20% by routine mammography. Cancers detected by mammography and CBE, but not those detected by breast self-exam, were much more likely to be early-stage. Detection by mammography increased with age, and a history of mammography use was associated with detection by mammography or CBE. Several commonly studied predictors of screening utilization in the US population were associated with CBE detection, but were less clearly related to or unrelated to mammography detection. Conclusion: Findings suggest that, during the 1990s in the US, most breast cancers among women under age 45, including those age 40-44, were self-detected. Few factors other than age and prior screening are verified predictors of method of breast cancer detection. C1 CDC, Div Canc Prevent & Control, Atlanta, GA 30341 USA. Emory Univ, Dept Biostat, Atlanta, GA 30322 USA. Columbia Univ, Sch Publ Hlth, Div Epidemiol, New York, NY 10032 USA. Fred Hutchinson Canc Res Ctr, Div Publ Hlth Sci, Seattle, WA 98104 USA. NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. Emory Univ, Dept Med, Atlanta, GA 30322 USA. RP Coates, RJ (reprint author), CDC, Div Canc Prevent & Control, K-55,4770 Buford Highway NE, Atlanta, GA 30341 USA. RI Brinton, Louise/G-7486-2015 OI Brinton, Louise/0000-0003-3853-8562 FU NCI NIH HHS [N01-CP 95604, N01-CP-95671, N01-CP-95672] NR 78 TC 27 Z9 27 U1 0 U2 0 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 JUN PY 2001 VL 12 IS 5 BP 431 EP 442 DI 10.1023/A:1011218005063 PG 12 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 455XA UT WOS:000170052000006 PM 11545458 ER PT J AU Sun, SY Yue, P Kelloff, GJ Steele, VE Lippman, SM Hong, WK Lotan, R AF Sun, SY Yue, P Kelloff, GJ Steele, VE Lippman, SM Hong, WK Lotan, R TI Identification of retinamides that are more potent than N-(4-hydroxyphenyl)retinamide in inhibiting growth and inducing apoptosis of human head and neck and lung cancer cells SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Article ID SYNTHETIC RETINOID FENRETINIDE; RECEPTOR-SELECTIVE RETINOIDS; CERVICAL-CARCINOMA CELLS; ABERRANT CRYPT FOCI; EPITHELIAL-CELLS; PROSTATE-CANCER; ACID; PROLIFERATION; MODULATION; INDUCTION AB The synthetic retinoid, N-(4-hydroxyphenyl)retinamide ( 4HPR), which is currently being evaluated in clinical trials for cancer prevention and therapy, inhibits the growth of a variety of malignant cells through induction of apoptosis, However, in the majority of tumor cells, this inhibitory effect of 4HPR requires high concentrations (>1 muM), which exceed the peak plasma level measured in humans. In the present study, we compared and contrasted the effects of several synthetic retinamides on the growth of human lung and head and neck cancer cells in vitro, We found that some retinamides, especially N-(2-carboxyphenyl)retinamide (2CPR), exhibited better growth inhibitory effects than 4HPR in some of the cell lines. 2CPR exerted potent growth inhibitory effects in 5 of 10 head and neck cancer cell lines and in 1 of 10 lung cancer cell lines (IC50, <0.8 muM). 2CPR (1 muM) induced apoptosis ranging from 10 to 60% in four of five cell lines, whereas 4HPR was ineffective at the same concentration. Unlike 4HPR, 2CPR (up to 10 muM) failed to induce reactive oxygen species production in these sensitive cell lines but could activate caspases 3 and 7 as well as increase poly(ADP-ribose)polymerase cleavage. Interestingly, the effect of 2CPR on cell growth could be suppressed by the specific retinoic acid receptor pan antagonist AGN193109. Our results suggest that 2CPR acts via retinoic acid receptors and may be a good candidate for prevention and treatment of some head and neck and lung cancers. C1 Univ Texas, MD Anderson Canc Ctr, Dept Thorac Head & Neck Med Oncol, Houston, TX 77030 USA. Univ Texas, MD Anderson Canc Ctr, Dept Clin Canc Prevent, Houston, TX 77030 USA. NCI, Div Canc Prevent, NIH, Bethesda, MD 20892 USA. RP Sun, SY (reprint author), Univ Texas, MD Anderson Canc Ctr, Dept Thorac Head & Neck Med Oncol, Box 432, Houston, TX 77030 USA. FU NCI NIH HHS [P01 CA52051, U19 CA68437]; NIDCR NIH HHS [P50 DE11906] NR 31 TC 25 Z9 27 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 JUN PY 2001 VL 10 IS 6 BP 595 EP 601 PG 7 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 442BU UT WOS:000169266000004 PM 11401908 ER PT J AU Anderson, KE Anderson, E Mink, PJ Hong, CP Kushi, LH Sellers, TA Lazovich, D Folsom, AR AF Anderson, KE Anderson, E Mink, PJ Hong, CP Kushi, LH Sellers, TA Lazovich, D Folsom, AR TI Diabetes and endometrial cancer in the Iowa women's health study SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Article ID BODY-FAT DISTRIBUTION; GROWTH-FACTOR-I; OLDER WOMEN; RISK-FACTORS; INSULIN; MELLITUS; HYPERINSULINEMIA; EPIDEMIOLOGY; POPULATION; MORTALITY AB Diabetes has been associated with increased risk of endometrial cancer in some epidemiological studies. Body mass index (BMI) and other measures of obesity have been associated positively with both diabetes and endometrial cancer. It is not clear whether or not the association of diabetes with endometrial cancer is explained entirely by obesity. Thus, we sought to test the hypothesis that diabetes is mot associated with endometrial cancer independent of obesity, We examined the association between self-reported diabetes (onset at > 30 years of age) and incident endometrial cancer in a prospective cohort study of 24,664 postmenopausal women in Iowa. Over 12 years of follow-up, 346 cases occurred among the cohort at risk. Data were analyzed using proportional hazards regression models. Diabetes was analyzed as reported at baseline and as a time-dependent variable using information obtained during follow-up. After adjustment for BMI, waist:hip ratio, and other covariates, the relative risk (RR) for women with diabetes versus women without diabetes was 1.43 [95% confidence interval (CI), 0.98-2.1]. The diabetes association was confined to women in the upper two BMI quintiles (RR, 1.47; 95% CI, 0.98-2.20), but a formal test of interaction was not statistically significant. Analyses that included diabetes ascertained at baseline and at follow-up gave similar results; the diabetes-associated RR in the higher BMI strata was 1.64 (95% CI, 1.16-2.31). We conclude that after adjustment for other risk factors, diabetes is associated with a modestly increased risk for endometrial cancer among women in this cohort. C1 Univ Minnesota, Div Epidemiol, Minneapolis, MN 55454 USA. Mayo Clin, Rochester, MN 55905 USA. Columbia Univ Teachers Coll, New York, NY 10027 USA. NCI, Environm Epidemiol Branch, Div Canc Epidemiol & Genet, Environm Epidemiol Branch, Rockville, MD 20852 USA. Univ Maryland, Sch Med, Dept Obstet & Gynecol, Baltimore, MD 21201 USA. RP Anderson, KE (reprint author), Univ Minnesota, Div Epidemiol, 1300 S 2nd St 300, Minneapolis, MN 55454 USA. OI Kushi, Lawrence/0000-0001-9136-1175 FU NCI NIH HHS [R01 CA-39742, T32CA09607] NR 47 TC 82 Z9 86 U1 1 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 JUN PY 2001 VL 10 IS 6 BP 611 EP 616 PG 6 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 442BU UT WOS:000169266000006 PM 11401910 ER PT J AU Stern, MC Umbach, DM Yu, MMC London, SJ Zhang, ZQ Taylor, JA AF Stern, MC Umbach, DM Yu, MMC London, SJ Zhang, ZQ Taylor, JA TI Hepatitis B, aflatoxin B-1, and p53 codon 249 mutation in hepatocellular carcinomas from Guangxi, People's Republic of China, and a meta-analysis of existing studies SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Article ID TUMOR-SUPPRESSOR GENE; VIRUS TRANSGENIC MICE; HOT-SPOT MUTATION; MOLECULAR PATHOGENESIS; RARE EVENT; OF-CHINA; EXPOSURE; HEPATOCARCINOGENESIS; HOTSPOT; LIVER AB The incidence of hepatocellular carcinomas (HCC) varies widely worldwide, with some of the highest incidence rates found in China. Chronic infection with the hepatitis B virus (HBV) and exposure to aflatoxins in foodstuffs are the main risk factors. A G to T transversion at codon 249 of the p53 gene (249(ser)) is commonly found in HCCs from patients in regions with dietary aflatoxin exposure. Because HBV infection is often endemic in high aflatoxin exposure areas, it is still unclear whether HBV acts as a confounder or as a synergistic partner in the development of the 249(ser) p53 mutation. Our report has two aims. First, we contribute data on HCCs from southern Guangxi, a high aflatoxin exposure area. Using DNA sequencing, we found that 36% (18 of 50) of tumors had a 249(ser) mutation. Also, 50% (30 of 60) were positive for p53 protein accumulation and 78% (28 of 36) were positive for HBV surface antigen, as detected by immunohistochemistry. Second, we present a metaanalysis, using our results along with those from 48 published studies, that examines the interrelationships among aflatoxin exposure, HBV infection, and p53 mutations in HCCs. We used a method that takes into account both within-study and study-to-study variability and found that the mean proportion of HCCs with the 249(ser) mutation was positively correlated with aflatoxin exposure (P = 0.0001). We found little evidence for an HBV-aflatoxin interaction modulating the presence of the p53 249(ser) mutation or any type of p53 mutation. C1 NIEHS, Mol Carcinogenesis Lab, NIH, Res Triangle Pk, NC 27709 USA. NIEHS, Epidemiol Branch, NIH, Res Triangle Pk, NC 27709 USA. NIEHS, Biostat Branch, NIH, Res Triangle Pk, NC 27709 USA. Univ So Calif, Keck Sch Med, Dept Prevent Med, Los Angeles, CA 90033 USA. Canc Inst Guangxi, Nanning 530021, Guangxi, Peoples R China. RP Taylor, JA (reprint author), NIEHS, Mol Carcinogenesis Lab, NIH, POB 12233,MD A3-05, Res Triangle Pk, NC 27709 USA. OI taylor, jack/0000-0001-5303-6398; London, Stephanie/0000-0003-4911-5290 FU NCI NIH HHS [R35 CA53890] NR 82 TC 60 Z9 67 U1 0 U2 3 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 JUN PY 2001 VL 10 IS 6 BP 617 EP 625 PG 9 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 442BU UT WOS:000169266000007 PM 11401911 ER PT J AU Poirier, LA Wise, CK Delongchamp, RR Sinha, R AF Poirier, LA Wise, CK Delongchamp, RR Sinha, R TI Blood determinations of S-adenosylmethionine, S-adenosylhomocysteine, and homocysteine: Correlations with diet SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Article ID MODERATE FOLATE-DEPLETION; ADENOSYL-L-METHIONINE; CHOLINE-DEVOID DIET; DNA METHYLATION; PLASMA HOMOCYSTEINE; METHYLENETETRAHYDROFOLATE REDUCTASE; SEX-DIFFERENCES; RENAL-FAILURE; BREAST-CANCER; COLON-CANCER AB An increasing number of both clinical and experimental studies have shown an association between deficiencies of the dietary sources of physiological methyl groups and cancer formation. The critical metabolic intermediate in a determination of methylation status is S-adenosylmethionine (SAM), the body's chief physiological methyl donor. The present study examined the erythrocyte levels of SAM and of its demethylated metabolite S-adenosylhomocysteine (SAH) in 66 normal subjects (33 men and 33 women), whose blood had been drawn at days 0, 7 and 14 of an experimental period during which they were fed a fixed diet. The plasma levels of homocysteine (HCys) were also determined in the same individuals at the same time points. In addition, the subjects had completed a food frequency questionnaire (FFQ) describing their usual dietary habits before being placed on the dietary regimen. The blood levels of SAM, SAH, and HCys were compared with the dietary intakes of folate, vitamin B-6, fats, and calories, both prior to using the FFQ and during the experimental period. The results indicated that the intraindividual differences were very low, but the interindividual differences were large for the values of SAM, SAH, SAM: SAH ratios, and HCys. Interestingly, the blood levels of SAM and HCys were higher in men than in women and generally showed the expected correlations with folate intake i.e., positive for SAM and negative for HCys. The intakes of folate (276 mug/days) and B-6 (1.87 mg/days) during the 2-week experimental period were relatively low compared with the usual intakes of these vitamins (375 and 2.06 mg/day for folate and B-6, respectively) but correlated well with each other during both periods of the study. Surprisingly, both men and women showed a significant rise in erythrocyte SAM:SAH ratios as a function of age. In addition, the combined results from men and women, even adjusted for gender, showed significant correlations between HCys and both weight and body mass index. On the other hand, during the experimental period of the study, blood SAM levels were inversely correlated with the intakes of both fat and calories when the data for both men and women were combined and adjusted for gender. The blood determinations of SAM and related compounds showed a high degree of reproducibility over time and thus appear to provide a practical marker of methylation status for the assessment of cancer risk from dietary, environmental, and genetic factors. C1 NCI, Environm Epidemiol Branch, Epidemiol & Biostat Program, NIH, Bethesda, MD 20892 USA. US FDA, Natl Ctr Toxicol Res, Div Mol Epidemiol, Jefferson, AR 72079 USA. RP Sinha, R (reprint author), NCI, Environm Epidemiol Branch, Epidemiol & Biostat Program, NIH, Execut Plaza S,6130 Execut Blvd, Bethesda, MD 20892 USA. RI Sinha, Rashmi/G-7446-2015 OI Sinha, Rashmi/0000-0002-2466-7462 FU NCI NIH HHS [Y01-CP3-0553] NR 67 TC 59 Z9 60 U1 0 U2 8 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 JUN PY 2001 VL 10 IS 6 BP 649 EP 655 PG 7 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 442BU UT WOS:000169266000011 PM 11401915 ER PT J AU Garcia-Closas, M Egan, KM Abruzzo, J Newcomb, PA Titus-Ernstoff, L Franklin, T Bender, PK Beck, JC Le Marchand, L Lum, A Alavanja, M Hayes, RB Rutter, J Buetow, K Brinton, LA Rothman, N AF Garcia-Closas, M Egan, KM Abruzzo, J Newcomb, PA Titus-Ernstoff, L Franklin, T Bender, PK Beck, JC Le Marchand, L Lum, A Alavanja, M Hayes, RB Rutter, J Buetow, K Brinton, LA Rothman, N TI Collection of genomic DNA from adults in epidemiological studies by buccal cytobrush and mouthwash SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Article ID TETRANUCLEOTIDE REPEAT POLYMORPHISM; PROSTATE-CANCER; CELL-DNA; GENE; POPULATIONS; MUTATIONS; SAMPLES; SWISH; SWABS; MEN AB Blood samples are an excellent source of large amounts of genomic DNA. However, alternative sources are often needed in epidemiological studies because of difficulties in obtaining blood samples. This report evaluates the buccal cytobrush and alcohol-containing mouthwash protocols for collecting DNA by mail. Several DNA extraction techniques are also evaluated. The study was conducted in two phases. In phase 1, we compared cytobrush and mouthwash samples collected by mail in two different epidemiological studies: (a) cytobrush samples (n = 120) from a United States case-control study of breast cancer; and (b) mouthwash samples (n = 40) from a prospective cohort of male United States farmers. Findings from phase 1 were confirmed in phase 2, where we randomized cytobrush (n = 28) and mouthwash (n = 25) samples among participants in the breast cancer study to directly compare both collection methods. The median human DNA yield determined by hybridization with a human DNA probe from phenol-chloroform extracts was 1.0 and 1.6 mug/2 brushes for phases 1 and 2, respectively, and 27.5 and 16.6 mug/mouthwash sample for phases 1 and 2, respectively. Most (94-100%) mouthwash extracts contained high molecular weight DNA (> 23 kb), in contrast to 55-61% of the brush extracts. PCR success rates for amplification of beta -globin gene fragments (268, 536, and 989 bp) were similar for cytobrush and mouthwash phenol-chloroform extracts (range, 94.4-100%). Also, we obtained high success rates in determining the number of CAG repeats in the androgen receptor gene, characterizing tetranucleotide microsatellites in six gene loci, and screening for mutations in the BRCA1/2 genes in a subset of phenol-chloroform DNA extracts. Relative to DNA extracted by phenol-chloroform from cytobrush samples, DNA extracted by NaOH had lower molecular weight, decreased PCR success rates for most assays performed, and unreliably high spectrophotometer readings for DNA yields. In conclusion, although DNA isolated from either mouthwash or cytobrush samples collected by mail from adults is adequate for a wide range of PCR-based assays, a single mouthwash sample provides substantially larger amounts and higher molecular weight DNA than two cytobrush samples. C1 NCI, Div Canc Epidemiol & Genet, Rockville, MD 20852 USA. Harvard Univ, Sch Publ Hlth, Boston, MA 02115 USA. Fred Hutchinson Canc Res Ctr, Seattle, WA 98104 USA. Univ Wisconsin, Madison, WI 53706 USA. Dartmouth Hitchcock Med Ctr, Lebanon, NH 03756 USA. Amer Type Culture Collect, Cell Biol, Manassas, VA 20110 USA. Coriell Inst, Camden, NJ 08103 USA. Univ Hawaii, Canc Res Ctr Hawaii, Honolulu, HI 96813 USA. RP Garcia-Closas, M (reprint author), NCI, Div Canc Epidemiol & Genet, 6120 Execut Blvd,EPS 7076, Rockville, MD 20852 USA. RI Garcia-Closas, Montserrat /F-3871-2015; Brinton, Louise/G-7486-2015; OI Garcia-Closas, Montserrat /0000-0003-1033-2650; Brinton, Louise/0000-0003-3853-8562; Rutter, Joni/0000-0002-6502-2361 FU NCI NIH HHS [CA47147, CA47305, CA67264, CA673338, CA69664] NR 24 TC 196 Z9 203 U1 4 U2 15 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 JUN PY 2001 VL 10 IS 6 BP 687 EP 696 PG 10 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 442BU UT WOS:000169266000016 PM 11401920 ER PT J AU Weinberg, DS Heyt, GJ Cavanagh, M Pitchon, D McGlynn, KA London, WT AF Weinberg, DS Heyt, GJ Cavanagh, M Pitchon, D McGlynn, KA London, WT TI Cholecystokinin and gastrin levels are not elevated in human pancreatic adenocarcinoma SO CANCER EPIDEMIOLOGY BIOMARKERS & PREVENTION LA English DT Article ID RECEPTOR; CANCER C1 Jefferson Med Coll, Div Gastroenterol & Hepatol, Dept Med, Philadelphia, PA 19107 USA. Univ Cincinnati, Dept Med, Cincinnati, OH 45221 USA. NIH, Epidemiol Branch, Bethesda, MD 20892 USA. Fox Chase Canc Ctr, Div Populat Sci, Philadelphia, PA 19111 USA. RP Weinberg, DS (reprint author), Jefferson Med Coll, Div Gastroenterol & Hepatol, Dept Med, 480 Main Bldg,132 S 10th St, Philadelphia, PA 19107 USA. NR 6 TC 6 Z9 6 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 JUN PY 2001 VL 10 IS 6 BP 721 EP 722 PG 2 WC Oncology; Public, Environmental & Occupational Health SC Oncology; Public, Environmental & Occupational Health GA 442BU UT WOS:000169266000022 PM 11401926 ER PT J AU Esa, A Coleman, AE Edelmann, P Silva, S Cremer, CG Janz, S AF Esa, A Coleman, AE Edelmann, P Silva, S Cremer, CG Janz, S TI Conformational differences in the 3-D nanostructure of the immunoglobulin heavy-chain locust a hotspot of chromosomal translocations in B lymphocytes SO CANCER GENETICS AND CYTOGENETICS LA English DT Article ID FLUORESCENCE MICROSCOPY; PARENTAL ORIGIN; C-MYC; ABERRATIONS; TERRITORIES; DNA AB Spectral precision distance microscopy was utilized to detect small but nonetheless consistently present conformational differences between the immunoglobulin heavy-chain gene clusters (IgH) that reside on the two chromosome 12 homologues in all diploid cells of the mouse. The euclidian distance (i.e.. the mean arithmetic three-dimensional [3-D] distance) between the 5 ' most IgH gene, C mu. and the 3 ' most IgH gene. C alpha. was used as the indicator to define the co-presence of a condensed IgH domain and a relaxed IgH domain in the same cell. In normal and malignant B cells in which IgH is actively rearranged and transcribed. the C mu /C alpha distance (genomic distance similar to 180 kb) was found to range from 87.5 to 121 nm on the condensed IgH domain and from 154 to 207 nm on the relaxed IgH domain. In non-B cells (fibroblasts. neutrophils. and macrophages), in which IgH is inactive, the C mu /C alpha distance was found to range from 136 to 154 nm on the condensed IgH domain and from 250 to 292 nm on the related IgH domain. These results suggested that conformational differences that may predispost: the relaxed IgH domain for illegitimate genetic recombinations. such as chromosomal translocations. are likely to exist in many cell types, including B cells. However. in B lymphocytes this structural predisposition may conspire with the lineage-specific ability to activate proto-oncogenes (after juxtaposition to IgH) to positively affect the preferential involvement of the relaxed IgH domain in chromosomal translocations. Additional studies are warranted to validate this working hypothesis. (C) 2001 Elsevier Science Inc. All rights reserved. C1 NCI, Div Basic Sci, Genet Lab, Bethesda, MD 20892 USA. Univ Heidelberg, Kirchhoff Inst Phys, Heidelberg, Germany. RP Janz, S (reprint author), NCI, Div Basic Sci, Genet Lab, Bldg 37,Room 2B10, Bethesda, MD 20892 USA. NR 16 TC 22 Z9 22 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 JUN PY 2001 VL 127 IS 2 BP 168 EP 173 DI 10.1016/S0165-4608(01)00455-1 PG 6 WC Oncology; Genetics & Heredity SC Oncology; Genetics & Heredity GA 445TB UT WOS:000169473600011 PM 11425458 ER PT J AU Blagosklonny, MV Pardee, AB AF Blagosklonny, MV Pardee, AB TI Exploiting cancer cell cycling for selective protection of normal cells SO CANCER RESEARCH LA English DT Review ID ANTICANCER DRUG TARGETS; TYROSINE KINASE INHIBITOR; DNA-DAMAGE; PROTEIN-KINASE; PHASE-II; PROSTATE-CANCER; MAMMALIAN-CELLS; CDK INHIBITORS; GROWTH-FACTORS; G(2) ARREST AB Chemotherapy of cancer is limited by its toxicity to normal cells. On the basis of discoveries in signal transduction and cell cycle regulation, novel mechanism-based therapeutics are being developed. Although these cell cycle modulators were designed to target cancer cells, some of them can also be applied for a different purpose, i.e., to protect normal cells against the lethality of chemotherapy. Loss of sensitivity of cancer cells to cell cycle inhibitors can be exploited for selective protection of normal cells that retain this response. Indeed, inhibition of redundant or overactivated pathways (e.g., growth factor-activated pathways) or stimulation of absent pathways in cancer cells (e.g, p53, Rb, and p16) may not arrest cycling of cancer cells. But growth arrest of normal cells will then permit selective killing of cancer cells by cycle-dependent chemotherapy. C1 NCI, Med Branch, NIH, Bethesda, MD 20892 USA. Dana Farber Canc Inst, Boston, MA 02115 USA. Harvard Univ, Sch Med, Boston, MA 02115 USA. RP Blagosklonny, MV (reprint author), NCI, Med Branch, NIH, Bldg 10,Room 12 N 226, Bethesda, MD 20892 USA. NR 99 TC 127 Z9 141 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 JUN 1 PY 2001 VL 61 IS 11 BP 4301 EP 4305 PG 5 WC Oncology SC Oncology GA 438KA UT WOS:000169051100002 PM 11389048 ER PT J AU Newbold, RR Banks, EP Bullock, B Jefferson, WN AF Newbold, RR Banks, EP Bullock, B Jefferson, WN TI Uterine adenocarcinoma in mice treated neonatally with genistein SO CANCER RESEARCH LA English DT Article ID BREAST-CANCER; EXPOSURE; DIETHYLSTILBESTROL; ESTROGENS; RATS AB The developing fetus is uniquely sensitive to perturbation with estrogenic chemicals. The carcinogenic effect of prenatal exposure to diethylstilbestrol (DES) is the classic example, Because phytoestrogen use in nutritional and pharmaceutical applications for infants and children is increasing, we investigated the carcinogenic potential of genistein, a naturally occurring plant estrogen in soy, in an experimental animal model previously reported to result in a high incidence of uterine adenocarcinoma after neonatal DES exposure. Outbred female CD-1 mice were treated on days 1-5 with equivalent estrogenic doses of DES (0.001 mg/kg/day) or genistein (50 mg/kg/day), At 18 months, the incidence of uterine adenocarcinoma was 35% for genistein and 31% for DES, These data suggest that genistein is carcinogenic if exposure occurs during critical periods of differentiation. Thus, the use of soy-based infant formulas in the absence of medical necessity and the marketing of soy products designed to appeal to children should be closely examined. C1 NIEHS, Dev Endocrinol Sect, Toxicol Lab, Environm Toxicol Program,Div Intramural Res, Res Triangle Pk, NC 27709 USA. Wake Forest Univ, Sch Med, Dept Pathol, Winston Salem, NC 27157 USA. RP Newbold, RR (reprint author), NIEHS, Dev Endocrinol Sect, Toxicol Lab, Environm Toxicol Program,Div Intramural Res, MD E4-02,POB 12233, Res Triangle Pk, NC 27709 USA. NR 19 TC 208 Z9 222 U1 0 U2 4 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 JUN 1 PY 2001 VL 61 IS 11 BP 4325 EP 4328 PG 4 WC Oncology SC Oncology GA 438KA UT WOS:000169051100007 PM 11389053 ER PT J AU Chokkalingam, AP McGlynn, KA Gao, YT Pollak, M Deng, J Sesterhenn, IA Mostofi, FK Fraumeni, JF Hsing, AW AF Chokkalingam, AP McGlynn, KA Gao, YT Pollak, M Deng, J Sesterhenn, IA Mostofi, FK Fraumeni, JF Hsing, AW TI Vitamin D receptor gene polymorphisms, insulin-like growth factors, and prostate cancer risk: A population-based case-control study in China SO CANCER RESEARCH LA English DT Article ID 1,25-DIHYDROXYVITAMIN D-3; BINDING-PROTEINS; ASSOCIATION; EXPRESSION; DENSITY; CELLS AB Operating through the vitamin D receptor (VDR), vitamin D inhibits prostate cancer growth and increases insulin-like growth factor binding protein (IGFBP) expression, suggesting that the vitamin D and insulin-like growth factor (IGF) regulatory systems may operate together to affect prostate cancer. Among 191 newly diagnosed prostate cancer cases and 304 randomly selected population controls in Shanghai, China, we found no significant association between the BsmI or FokI VDR gene polymorphisms and prostate cancer risk. However, we found that among men with the ff FokI genotype, those in the highest tertile of plasma IGFBP-3 had a decreased risk versus those in the lowest tertile (odds ratio, 0.14; 95% confidence interval, 0.04-0.56; P-trend < 0.01), whereas among men with the FF and Ff genotypes, IGFBP-3 was not associated with risk. Similarly, IGFBP-1 was inversely associated with prostate cancer risk only among men with the ff FokI genotype (odds ratio, 0.25; 95% confidence interval, 0.07-0.85; P-trend = 0.02). No such FokI genotype-specific effects were observed for IGF-I or IGF-II. Our findings in a low-risk population suggest that the IGF and vitamin D regulatory systems may interact to affect prostate cancer risk. Larger studies are needed to confirm these findings and clarify the underlying mechanisms. C1 NCI, Div Canc Epidemiol & Genet, NIH, Bethesda, MD 20892 USA. Univ Maryland, Dept Epidemiol & Prevent Med, Baltimore, MD 21201 USA. Shanghai Canc Inst, Shanghai 200032, Peoples R China. Sir Mortimer B Davis Jewish Hosp, Canc Prevent Res Unit, Montreal, PQ H3T 1E2, Canada. McGill Univ, Dept Oncol, Montreal, PQ H3T 1E2, Canada. Armed Forces Inst Pathol, Washington, DC 20306 USA. RP Chokkalingam, AP (reprint author), NCI, Div Canc Epidemiol & Genet, NIH, EPS-MSC-7234,6120 Execut Blvd, Bethesda, MD 20892 USA. RI Pollak, Michael/G-9094-2011 OI Pollak, Michael/0000-0003-3047-0604 NR 20 TC 65 Z9 69 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD JUN 1 PY 2001 VL 61 IS 11 BP 4333 EP 4336 PG 4 WC Oncology SC Oncology GA 438KA UT WOS:000169051100009 PM 11389055 ER PT J AU Vahakangas, KH Bennett, WP Castren, K Welsh, JA Khan, MA Blomeke, B Alavanja, MCR Harris, CC AF Vahakangas, KH Bennett, WP Castren, K Welsh, JA Khan, MA Blomeke, B Alavanja, MCR Harris, CC TI p53 and K-ras mutations in lung cancers from former and never-smoking women SO CANCER RESEARCH LA English DT Article ID ENVIRONMENTAL TOBACCO-SMOKE; TUMOR-SUPPRESSOR GENES; RESIDENTIAL RADON EXPOSURE; SQUAMOUS-CELL CARCINOMAS; NONSMOKING WOMEN; ONCOGENE ACTIVATION; MOLECULAR EPIDEMIOLOGY; PASSIVE SMOKING; TRANSGENIC MICE; POINT MUTATIONS AB Somatic p53 mutations are common in lung cancer. Active cigarette smoking is positively correlated with the total frequency of p53 mutations and G:C to T:A transversions on the nontranscribed (DNA coding) strand, Mutational hotspots within the p53 gene, e,g,, codon 157, have been identified for tobacco-related lung cancer, whereas these same mutations are found rarely in other cancers. Such data implicate specific p53 mutations as molecular markers of smoking, Because limited data exist concerning the p53 mutation frequency and spectra in ex-smokers and nonsmokers, rye have analyzed p53 and K-rns mutations in 126 lung cancers from a population-based case-control study of nonsmoking (n = 117) or ex-smoking (n = 9) women from Missouri with quantitative assessments of exposure to environmental tobacco smoke. Mutations in the p53 gene mere found in lung cancers from lifetime nonsmokers (19%) and ex-smokers (67%; odds ratio, 9.08; 95% confidence interval, 2.06-39.98). AII deletions were found in tumors from patients who mere either ex-smokers or nonsmokers exposed to passive smoking. The G:C to A:T transitions (11 of 28; 39%) were the most frequent p53 mutations found and clustered in tumors from lifetime nonsmokers without passive smoke exposure. The incidence of K-ras codon 12 or 13 mutations was 11% (14 of 115 analyzed) with no difference between long-term ex-smokers and nonsmokers, These and other results indicate that p53 mutations occur more commonly in smokers and ex-smokers than in never-smokers. Such comparisons provide additional evidence of genetic damage caused by tobacco smoke during lung carcinogenesis. C1 NCI, Human Carcinogenesis Lab, NIH, Bethesda, MD 20892 USA. NCI, Div Canc Epidemiol & Genet, Bethesda, MD 20892 USA. City Hope Beckman Res Inst, Div Mol Med, Duarte, CA 91010 USA. Univ Oulu, Dept Pharmacol & Toxicol, FIN-90014 Oulu, Finland. RP Harris, CC (reprint author), NCI, Human Carcinogenesis Lab, NIH, Bldg 37,Room 2C05,MSC 4255, Bethesda, MD 20892 USA. NR 75 TC 84 Z9 95 U1 2 U2 7 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 JUN 1 PY 2001 VL 61 IS 11 BP 4350 EP 4356 PG 7 WC Oncology SC Oncology GA 438KA UT WOS:000169051100013 PM 11389059 ER PT J AU Krivak, TC McBroom, JW Seidman, J Venzon, D Crothers, B MacKoul, PJ Rose, GS Carlson, JW Birrer, MJ AF Krivak, TC McBroom, JW Seidman, J Venzon, D Crothers, B MacKoul, PJ Rose, GS Carlson, JW Birrer, MJ TI Abnormal fragile histidine triad (FHIT) expression in advanced cervical carcinoma: A poor prognostic factor SO CANCER RESEARCH LA English DT Article ID SQUAMOUS-CELL CARCINOMA; PROTEIN EXPRESSION; UTERINE CERVIX; CLINICOPATHOLOGICAL SIGNIFICANCE; PREMALIGNANT LESIONS; CHROMOSOME 3P; GENE; CANCER; SURVIVAL; DISEASE AB The FHIT gene is a candidate tumor suppressor gene that has been implicated in the development of cervical carcinoma. We hypothesized that abnormal Fhit expression might be a poor prognostic factor for patients with cervical cancer. The tumors from 59 high-risk patients (stage ii-in) were evaluated for abnormal Fhit expression by immunohistochemical staining. Abnormal Fhit expression (absent or reduced) was noted in 66% of the specimens, There was no statistical difference with respect to stage, performance status, para-aortic node metastasis, completion of therapy, grade, race, age, and HIV status between the normal and abnormal Fhit expression groups, The 3-year survival for patients whose tumors displayed normal Fhit expression versus abnormal Fhit expression was 74% versus 37%, respectively, Univariate analysis demonstrated a difference in survival that was statistically significant for age < 55 years versus greater than or equal to 55 years (P = 0.015), normal Fhit expression versus abnormal Fhit expression (P = 0.015), and stage II versus stage III (P = 0.033), Multivariate analysis showed that abnormal Fhit expression was a poor prognostic factor (P = 0.015). C1 NCI, Div Clin Sci, Dept Cell & Canc Biol, Rockville, MD 20850 USA. Washington Hosp Ctr, Washington, DC 20307 USA. Walter Reed Army Med Ctr, Washington, DC 20307 USA. RP Birrer, MJ (reprint author), Key W Res Ctr, 9610 Med Ctr Dr,Room 300, Rockville, MD 20850 USA. RI Venzon, David/B-3078-2008 NR 30 TC 17 Z9 22 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD JUN 1 PY 2001 VL 61 IS 11 BP 4382 EP 4385 PG 4 WC Oncology SC Oncology GA 438KA UT WOS:000169051100018 PM 11389064 ER PT J AU Fan, MY Goodwin, ME Birrer, MJ Chambers, TC AF Fan, MY Goodwin, ME Birrer, MJ Chambers, TC TI The c-Jun NH2-terminal protein kinase/AP-1 pathway is required for efficient apoptosis induced by vinblastine SO CANCER RESEARCH LA English DT Article ID STRESS-INDUCED APOPTOSIS; CELL-CYCLE PROGRESSION; FACTOR-ALPHA PROMOTER; CARCINOMA-CELLS; P53-INDEPENDENT APOPTOSIS; DNA REREPLICATION; CANCER CELLS; DEATH; PHOSPHORYLATION; JNK AB Vinblastine is an important antitumor agent that induces G(2)-M arrest and subsequent apoptosis in a wide variety of cell Lines, but the molecular mechanisms that link mitotic arrest and apoptosis are poorly understood. The AP-1 transcription factor has been implicated in many critical cellular processes, including apoptosis, and is a major target of the c-Jun NH2-terminal kinase signaling pathway that is activated by vinblastine and other microtubule inhibitors. In this study we sought to determine the role of c-Jun NH2-terminal kinase/AP-1 in the response of KB3 carcinoma cells to vinblastine, For this purpose, we generated RB3 cell lines that stably expressed the c-Jun dominant-negative deletional mutant TAM67, which lacks the NH2-terminal transactivation domain. KB3-TAM67 cell lines displayed normal growth kineties and essentially unaltered basal AP-1 activity, but vinblastine-induced phosphorylation of c-Jun and activating transcription factor-2, and AP-1 activation, were strongly inhibited. KB3-TAM67 cell lines arrested normally at G(2)-M in response to vinblastine, but were significantly more resistant to the drug, exhibiting markedly delayed apoptosis and increased overall survival, relative to control cells. To investigate the underlying mechanisms, differential expression of apoptotic regulatory genes was monitored by immunoblot and cDNA microarray analysis, We found that vinblastine treatment caused down-regulation of p53 and its target p21 and up-regulation of tumor necrosis factor or, Bak, and several ether genes in control but not in KB3-TAM67 cells, identifying these genes as putative targets of vinblastine-inducible AP-1. These results demonstrate that vinblastine-inducible AP-1 plays a destructive, proapoptotic role and may do so by regulating the expression of a specific subset of target genes that promotes efficient apoptotic cell death following mitotic arrest. C1 Univ Arkansas Med Sci, Dept Biochem & Mol Biol, Little Rock, AR 72205 USA. NCI, Dept Cell & Canc Biol, Med Branch, NIH, Bethesda, MD 20850 USA. RP Chambers, TC (reprint author), Univ Arkansas Med Sci, Dept Biochem & Mol Biol, Mail Slot 516,4301 W Markham St, Little Rock, AR 72205 USA. FU NCI NIH HHS [CA-75577] NR 61 TC 90 Z9 101 U1 0 U2 5 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD JUN 1 PY 2001 VL 61 IS 11 BP 4450 EP 4458 PG 9 WC Oncology SC Oncology GA 438KA UT WOS:000169051100029 PM 11389075 ER PT J AU Lee, FT Rigopoulos, A Hall, C Clarke, K Cody, SH Smyth, FE Liu, ZQ Brechbiel, MW Hanai, N Nice, EC Catimel, B Burgess, AW Welt, S Ritter, G Old, LJ Scott, AM AF Lee, FT Rigopoulos, A Hall, C Clarke, K Cody, SH Smyth, FE Liu, ZQ Brechbiel, MW Hanai, N Nice, EC Catimel, B Burgess, AW Welt, S Ritter, G Old, LJ Scott, AM TI Specific localization, gamma camera imaging, and intracellular trafficking of radiolabelled chimeric anti-G(D3) ganglioside monoclonal antibody KM871 in SK-MEL-28 melanoma xenografts SO CANCER RESEARCH LA English DT Article ID PHASE-I TRIAL; METASTATIC MELANOMA; GD3 GANGLIOSIDE; MALIGNANT-MELANOMA; ANTIGENIC SYSTEMS; NORMAL TISSUE; MOUSE; SURFACE; TUMOR; BINDING AB The chimeric monoclonal antibody KM871, directed against the G(D3) antigen, is under evaluation for its potential to target melanoma, To facilitate the in vivo evaluation of biodistribution properties and measurement of pharmacokinetics. KM871 was radiolabeled with I-125 via tyrosine residues and with In-111 via the bifunctional metal ion chelator C-functionalized hairs-cyclohexyl diethylenetriaminepentaacetic acid (CHX-A " -DTPA) to lysine residues, Using antigen-positive SK-MEL-28 melanoma cells, immunoreactivities of 42 and 40% cell binding were obtained, respectively, for the two radioconjugates, Binding was enhanced in the presence of added unlabeled antibody, A humanized A33 antibody was similarly labeled with the two isotopes and used as a control. To determine and compare in vivo biodistribution characteristics of KM871 radiolabeled with In-111 or I-125, mixtures of the radioconjugates were injected i.v. into BALB/c nude mice bearing GD(3)-positive-SK-MEL-28 melanoma xenografts, Gamma camera images were acquired; groups of five mice were sacrificed at various time intervals, and tumors, blood, and tissues were analyzed. In-111-labeIed CHX-A " -DTPA-KM871 showed a maximum tumor uptake of 41.9 +/- 7.0% injected dose/g at 72 h with prolonged retention over a 15-day period, The tumor:blood ratio was 3:1 by 72 h, and higher ratios were observed at later time points. No abnormal accumulation of In-111-Iabeled conjugate was found in normal tissues, In contrast, there was little accumulation of I-125-labeled KM871 in the same tumors. The specificity of antibody localization was confirmed by the low tumor uptake values for radiolabeled control antibody. Gamma camera imaging demonstrated excellent uptake of In-111-labeled CHX-A " -DTPA-KM871 in the xenografts, Chromatographic analyses of xenograft cytosolic extracts demonstrated tumor internalization and catabolism of radiolabeled KM871 with the formation of small molecular weight metabolites, Laser scanning confocal microscopy demonstrated that the majority of intracellular KM871 is localized to lysosomes, Despite the catabolism of the radioconjugate, a dose-dependent increase in KM871 tumor localization was shown through immunohistochemical examination of xenograft biopsies. This study demonstrates for the first time the in vivo localization of a radiolabeled anti-G(D3), monoclonal antibody to G(D3) expressing xenografts using gamma camera scanning techniques and tumor cell internalization of KM871 tagged with a green fluorescent dye, Alexa Fluor 488, through confocal microscopy, KM871 has potential for targeting tumors in patients with melanoma. C1 Austrian & Repatriat Med Ctr, Ludwig Inst Canc Res, Melbourne Branch, Heidelberg, Vic 3084, Australia. NCI, Radimmune & Inorgan Chem Sect, Radiat Oncol Branch, NIH, Bethesda, MD 20892 USA. Kyowa Hakko Kogyo Co Ltd, Tokyo Res Labs, Tokyo 194, Japan. Mem Sloan Kettering Canc Ctr, Ludwig Inst Canc Res, New York, NY 10021 USA. Austrian & Repatriat Med Ctr, Dept Nucl Med, Heidelberg, Vic 3084, Australia. Austrian & Repatriat Med Ctr, Ctr Positron Emiss Tomog, Heidelberg, Vic 3084, Australia. RP Lee, FT (reprint author), Austrian & Repatriat Med Ctr, Ludwig Inst Canc Res, Melbourne Branch, Studley Rd, Heidelberg, Vic 3084, Australia. RI Nice, Edouard/B-1026-2011 NR 32 TC 26 Z9 27 U1 1 U2 4 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 JUN 1 PY 2001 VL 61 IS 11 BP 4474 EP 4482 PG 9 WC Oncology SC Oncology GA 438KA UT WOS:000169051100032 PM 11389078 ER PT J AU Grosenbach, DW Barrientos, JC Schlom, J Hodge, JW AF Grosenbach, DW Barrientos, JC Schlom, J Hodge, JW TI Synergy of vaccine strategies to amplify antigen-specific immune responses and antitumor effects SO CANCER RESEARCH LA English DT Article ID HUMAN CARCINOEMBRYONIC ANTIGEN; COLONY-STIMULATING FACTOR; TUMOR-ASSOCIATED ANTIGEN; T-CELL RESPONSES; RECOMBINANT ANTICANCER VACCINES; COSTIMULATORY MOLECULE B7; CANCER-PATIENTS; PHASE-I; GM-CSF; THERAPEUTIC EFFECTIVENESS AB Several different vaccine strategies have been evaluated and combined in an attempt to amplify T-cell responses toward induction of antitumor immunity. The model tumor antigen used was carcinoembryonic antigen (CEA), While initial T-cell activation studies were conducted in conventional mice, combined vaccine strategy studies and antitumor studies were conducted in transgenic mice in which CEA is expressed in normal gastrointestinal tissue and CEA protein is found in sera. The studies reported here demonstrate: (a) A recombinant avipox (fowlpox, rF) vector expressing the signal 1 (CEA) and the B7-1 costimulatory molecule transgenes (designated rF-CEA/B7-1) is more potent in inducing CEA-specific T-cell responses than rF-CEA; one administration of recombinant fowlpox vector expressing CEA and three different costimulatory molecule transgenes (B7-1, ICAM-1, LFA-3, designated rF-CEA/TRICOM) was more potent in inducing CEA-specific T-cell responses than four vaccinations with rF-CEA or two vaccinations with rF-CEA/B7-1, Moreover, up to four vaccinations with rF-CEA/TRICOM induced greater CEA-specific T-cell responses with each vaccination. (b) A diversified prime and boost strategy using a prime with a recombinant vaccinia vector expressing CEA and the triad of costimulatory molecules (designated rV-CEA/TRICOM) and a boost with rP-CEA/TRICOM was more potent in inducing CEA-specific T-cell responses than the repeated use of rF-CEA/TRICOM alone. (c) The addition of granulocyte macrophage colony-stimulating factor (GM-CSF) to the rF-CEA or rF-CEA/TRICOM vaccinations via the simultaneous administration of a rF-GM-CSF vector enhanced CEA-specific T-cell responses, These strategics (TRICOM/diversified prime and boost/GM-CSF) were combined to treat CEA-expressing carcinoma Liver metastases in CEA-transgenic mice; vaccination was initiated 14 days posttumor transplant. Antitumor effects in terms of survival and CD8(+) and CD4(+) responses specific for CEA were also observed in this CEA-transgenic mouse model. These studies demonstrate that the use of cytokines and diversified prime and boost regimens can be combined with the use of recombinant vectors expressing signal 1 and multiple costimulatory molecules to further amplify T-cell responses toward more effective vaccine strategies. C1 NCI, Tumor Immunol & Biol Lab, NIH, Bethesda, MD 20892 USA. NIH, Howard Hughes Med Inst, Bethesda, MD 20892 USA. RP Schlom, J (reprint author), NCI, Tumor Immunol & Biol Lab, NIH, 10 Ctr Dr,Room 8B09, Bethesda, MD 20892 USA. RI Hodge, James/D-5518-2015 OI Hodge, James/0000-0001-5282-3154 NR 72 TC 119 Z9 121 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 JUN 1 PY 2001 VL 61 IS 11 BP 4497 EP 4505 PG 9 WC Oncology SC Oncology GA 438KA UT WOS:000169051100035 PM 11389081 ER PT J AU Nishizuka, S Tsujimoto, H Stanbridge, EJ AF Nishizuka, S Tsujimoto, H Stanbridge, EJ TI Detection of differentially expressed genes in HeLa x fibroblast hybrids using subtractive suppression hybridization SO CANCER RESEARCH LA English DT Article ID HUMAN CELL HYBRIDS; CYCLE ARREST; IDENTIFICATION; TUMORIGENICITY; CDNA; TRANSFORMATION; CHROMOSOME-11; CARCINOMA; DELETION; CLONING AB To understand genetic differences and similarities between tumorigenic and nontumorigenic HeLa x fibroblast hybrid cells, subtractive suppression hybridization (SSH), based on suppression PCR and a combination of normalization and subtraction in a single procedure, was used. Using the nontumorigenic CGL1 and tumorigenic CGL3, forward (CGL1-CGL3) and reverse (CGL3-CGL1) subtracted libraries mere constructed. Among 192 clones, seven were identified as differentially expressed genes specific for either CGL1 or CGL3. All seven were not reported previously as differentially expressed genes in this hybrid system, In the forward subtraction, p16 was isolated, indicating the involvement of the loss of tumorigenic phenotype, Subsequent transfection of wild-type p16 to the tumorigenic CGL3 showed growth suppression in colony formation assay; however, no tumor suppression was observed when the transfectant was inoculated into nude mice. These results indicate that: (a) SSH is a suitable method to identify differentially expressed genes in two types of cells; and (b) although p16 plays some roles in growth suppression, the p16-transfected CGL3 is still capable to proliferate in vivo. C1 Univ Calif Irvine, Coll Med, Dept Microbiol & Mol Genet, Irvine, CA 92697 USA. RP Nishizuka, S (reprint author), NCI, Mol Pharmacol Lab, NIH, Bldg 37,Room 4C-09,9000 Rockville Pike, Bethesda, MD 20892 USA. FU NCI NIH HHS [CA 19104] NR 27 TC 11 Z9 15 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD JUN 1 PY 2001 VL 61 IS 11 BP 4536 EP 4540 PG 5 WC Oncology SC Oncology GA 438KA UT WOS:000169051100040 PM 11389086 ER PT J AU Alt, A Ohba, M Li, LW Gartsbein, M Belanger, A Denning, MF Kuroki, T Yuspa, SH Tennenbaum, T AF Alt, A Ohba, M Li, LW Gartsbein, M Belanger, A Denning, MF Kuroki, T Yuspa, SH Tennenbaum, T TI Protein kinase C delta-mediated phosphorylation of alpha 6 beta 4 is associated with reduced integrin localization to the hemidesmosome and decreased keratinocyte attachment SO CANCER RESEARCH LA English DT Article ID EPIDERMAL GROWTH-FACTOR; SIGNAL-TRANSDUCTION; TYROSINE PHOSPHORYLATION; CYTOPLASMIC DOMAIN; ALPHA(6)BETA(4) INTEGRIN; DOWN-REGULATION; CELL-ADHESION; TERMINAL DIFFERENTIATION; MOUSE KERATINOCYTES; BASEMENT-MEMBRANE AB In mammalian epidermis, expression of the alpha6 beta4 integrin is restricted to the hemidesmosome complexes, which connect the proliferative basal cell layer with the underlying basement membrane. Keratinocyte differentiation is associated with down-regulation of alpha6 beta4 expression and detachment of keratinocytes from the basement membrane. Neoplastic keratinocytes delay maturation, proliferate suprabasally, and retain the expression of the alpha6 beta4 integrin in suprabasal cells disassociated from the hemidesmosomes. We now show that the alpha6 beta4 integrin is a substrate for serine phosphorylation by protein kinase C in keratinocytes. Furthermore, protein kinase C-mediated phosphorylation of alpha6 beta4 is associated with redistribution of this integrin from the hemidesmosome to the cytosol. Specifically, in vitro kinase assays identified the protein kinase C delta as the primary isoform phosphorylating alpha6 and beta4 integrin subunits, Using recombinant protein kinase C adenoviruses, overexpression of protein kinase C delta but not protein kinase C alpha in primary keratinocytes increased beta4 serine phosphorylation, decreased alpha6 beta4 localization to the hemidesmosome complexes, and reduced keratinocyte attachment. Taken together, these results establish a link between protein kinase C delta -mediated serine phosphorylation of alpha6 beta4 integrin and its effects on alpha6 beta4 subcellular localization and keratinocyte attachment to the laminin underlying matrix. C1 Bar Ilan Univ, Fac Life Sci, IL-52900 Ramat Gan, Israel. NCI, Cellular Carcinogenesis & Tumor Promot Lab, NIH, Bethesda, MD 20892 USA. Showa Univ, Inst Mol Oncol, Tokyo 1428555, Japan. RP Tennenbaum, T (reprint author), Bar Ilan Univ, Fac Life Sci, IL-52900 Ramat Gan, Israel. RI Kuroki, Toshio/A-9500-2011 OI Kuroki, Toshio/0000-0001-6369-4351 NR 76 TC 35 Z9 35 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 0008-5472 J9 CANCER RES JI Cancer Res. PD JUN 1 PY 2001 VL 61 IS 11 BP 4591 EP 4598 PG 8 WC Oncology SC Oncology GA 438KA UT WOS:000169051100049 PM 11389095 ER PT J AU Mandil, R Ashkenazi, E Blass, M Kronfeld, I Kazimirsky, G Rosenthal, G Umansky, F Lorenzo, PS Blumberg, PM Brodie, C AF Mandil, R Ashkenazi, E Blass, M Kronfeld, I Kazimirsky, G Rosenthal, G Umansky, F Lorenzo, PS Blumberg, PM Brodie, C TI Protein kinase C alpha and protein kinase C delta play opposite roles in the proliferation and apoptosis of glioma cells SO CANCER RESEARCH LA English DT Article ID FIBRILLARY ACIDIC PROTEIN; HUMAN ASTROCYTOMA-CELLS; SIGNAL-TRANSDUCTION; PROTEOLYTIC ACTIVATION; MALIGNANT GLIOMAS; PHORBOL ESTER; GLUTAMINE-SYNTHETASE; COMPLEMENTARY-DNA; GLIAL-CELLS; GROWTH-RATE AB Protein kinase C (PRC) has been implicated in the proliferation and apoptosis of glial tumors, but the role of specific PKC isoforms remains unresolved, Comparing brain tumors differing in degree of malignancy, me found that malignant gliomas expressed higher levels of PKC alpha and lower levels of PKC delta as compared with low-grade astrocytomas, Consistent with a mechanistic role for these differences, overexpression of PKC alpha in the human U87 glioma cell Line resulted in enhanced cell proliferation and decreased glial fibrillary acidic protein (GFAP) expression as compared with controls, Reciprocally, overexpression of PKC delta inhibited cell proliferation and enhanced GFAP expression, Using PKC chimeras, me found that the regulatory domains of PKC alpha and PKC delta mediated their effects on cell proliferation and GFAP expression, PKC alpha and delta have been implicated as potential signaling molecules in apoptosis, Therefore, we examined the role of these isoforms in the resistance of glioma cells to apoptotic stimuli, In U87 cells, manipulation of PKC alpha levers had little effect on apoptosis in response to etoposide. In contrast, overexpression of PKC delta rendered the U87 cells more sensitive to the apoptotic effect of etoposide, and PKC delta was cleaved in these cells by a caspase-dependent process. Furthermore, the glioma cell line U373, which expresses endogenous PKC delta, underwent apoptosis in response to etoposide, and the apoptotic response was blocked by the PKC delta inhibitor rottlerin, Our results suggest that PKC alpha and PKC delta play opposite roles in the proliferation and apoptosis of glioma cells. C1 Bar Ilan Univ, Fac Life Sci, IL-52900 Ramat Gan, Israel. Hadassa Med Ctr, Dept Neurosurg, Jerusalem, Israel. NCI, Cellular Carcinogenesis & Tumor Promot Lab, NIH, Bethesda, MD 20892 USA. RP Brodie, C (reprint author), Bar Ilan Univ, Fac Life Sci, IL-52900 Ramat Gan, Israel. NR 49 TC 142 Z9 148 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 JUN 1 PY 2001 VL 61 IS 11 BP 4612 EP 4619 PG 8 WC Oncology SC Oncology GA 438KA UT WOS:000169051100052 PM 11389098 ER PT J AU Blagosklonny, MV Giannakakou, P Romanova, LY Ryan, KM Vousden, KH Fojo, T AF Blagosklonny, MV Giannakakou, P Romanova, LY Ryan, KM Vousden, KH Fojo, T TI Inhibition of HIF-1-and wild-type p53-stimulated transcription by codon Arg175 p53 mutants with selective loss of functions SO CARCINOGENESIS LA English DT Article ID HYPOXIA-INDUCIBLE FACTOR-1; TUMOR-SUPPRESSOR P53; CELL-CYCLE ARREST; P53-MEDIATED TRANSACTIVATION; COLORECTAL TUMORIGENESIS; GENE-EXPRESSION; FACTOR 1-ALPHA; HUMAN CANCER; MUTATIONS; APOPTOSIS AB Overexpression of ectopic mutant p53 represses wild-type p53-stimulated transcription, known as a dominant negative effect. On the other hand, overexpression of wildtype p53 can repress transcription stimulated by several transcription factors, including hypoxia-inducible factor-1 (HIF-1). Using a panel of well-characterized Arg175 p53 mutants we found that only mutants (Tyr175, Trp175, Asp175 and Phe175) which have completely lost their ability to transactivate repress wild-type p53-stimulated Bax, p21 and PG13 promoter constructs. In contrast, Asn175, Gln175, Leu175 and Pro175 mutants which partially retained transactivating functions did not exert dominant negative effects against PG13 and p21 promoter constructs. However, these latter mutants failed to activate Bax and, instead, exerted a dominant negative effect on a Bax-Luc promoter construct. We conclude that a dominant negative effect is promoter selective as a consequence of selective loss of transactivating function. Albeit less potent than wild-type p53, all Arg175 p53 mutants retained partial ability to repress HIF-1-stimulated transcription. We propose that transrepression and the dominant negative effect have similar mechanisms and may involve competition with transcription factors (wild-type p53, HIF-1, etc.) for cofactors such as p300. Thus, a p53(22/23) mutant, which is deficient in p300 binding, did not exert dominant negative effects. Like transrepression, the dominant negative effect required overexpression of mutant p53 and, therefore, is not dominant. In the presence of a wild-type p53 allele, levels of endogenous mutant p53 protein were low in heterozygous cells. Endogenous mutant p53 became overexpressed only after loss of the second p53 allele. Therefore, endogenous mutant p53s are unable to display a dominant negative effect. This explains why loss of the second p53 allele is required to eliminate p53 functions in cancer cells. C1 NCI, Med Branch, NIH, Bethesda, MD 20892 USA. NCI, Genet Lab, NIH, Bethesda, MD 20892 USA. NCI, Frederick Canc Res & Dev Ctr, ABL Basic Res Program, Frederick, MD 21702 USA. RP Blagosklonny, MV (reprint author), NCI, Med Branch, NIH, Bldg 10, Bethesda, MD 20892 USA. NR 58 TC 28 Z9 30 U1 1 U2 3 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0143-3334 J9 CARCINOGENESIS JI Carcinogenesis PD JUN PY 2001 VL 22 IS 6 BP 861 EP 867 DI 10.1093/carcin/22.6.861 PG 7 WC Oncology SC Oncology GA 440JE UT WOS:000169171500003 PM 11375890 ER PT J AU Kumar, S Chang, RL Wood, AW Xie, JG Huang, MT Cui, XX Kole, PL Sikka, HC Balani, SK Conney, AH Jerina, DM AF Kumar, S Chang, RL Wood, AW Xie, JG Huang, MT Cui, XX Kole, PL Sikka, HC Balani, SK Conney, AH Jerina, DM TI Tumorigenicity of racemic and optically pure bay region diol epoxides and other derivatives of the nitrogen heterocycle dibenz[a,h]acridine on mouse skin SO CARCINOGENESIS LA English DT Article ID MAMMALIAN-CELL MUTAGENICITY; RAT-LIVER MICROSOMES; NEWBORN MICE; METABOLISM; DIHYDRODIOLS; REACTIVITY; BACTERIAL; 3,4-DIOL-1,2-EPOXIDES; DIBENZACRIDINE; BENZACRIDINE AB CD-1 female mice were initiated with a single topical application of 500 mmol dibenz[a,h]acridine (DB[a,h]Acr), its racemic trans-1,2-, 3,4-, 8,9- and 10,11-dihydrodiols, racemic DB[a,h]Acr 3,4-diol 1,2-epoxide-1 and -2 or racemic DB[a,h]Acr 10,11-diol 8,9-epoxide-1 and -2, where the benzylic hydroxyl group is either cis (isomer 1) or trans (isomer 2) to the epoxide oxygen. The mice were subsequently treated twice weekly with 12-O-tetradecanoyl-phorbol 13-acetate for 25 weeks. High tumorigenicity was observed only for DB[a,h]Acr, its 10,11-dihydrodiol and DB[a,h]Acr 10,11-diol 8,9-epoxide-2 (3.3, 1.2 and 1.6 tumors/mouse, respectively). The tumor-initiating activity of a 50 nmol dose of DB[a,h]Acr and the optically active (+)- and (-)-enantiomers of DB[a,h]Acr 10,11-dihydrodiol and of the optically active DB[a,h]Acr 10,11 diol 8,9-epoxide-1 and -2 were also studied. Only DB[a,h]Acr, (-)DB[a,h]Acr (10R,11R)-dihydrodiol and the bay region(+)-(8R,9S,10S,11R)-diol epoxide-2 were highly active (1.6, 1.7 and 2.4 tumors/mouse, respectively). These results are consistent with previous studies which showed that the corresponding bay region RSSR diol epoxides of benzo[a]pyrene, benz[a]anthracene, chrysene and benzo[c]phenanthrene as well as the aza-polycyclic dibenz[c,h]acridine are the most tumorigenic isomers. C1 Rutgers State Univ, Coll Pharm, Canc Res Lab, Piscataway, NJ 08854 USA. SUNY Coll Buffalo, Great Lakes Ctr, Environm Toxicol & Chem Lab, Buffalo, NY 14222 USA. Hoffmann La Roche Inc, Roche Res Ctr, Nutley, NJ 07110 USA. NIDDKD, NIH, Bethesda, MD 20892 USA. RP Chang, RL (reprint author), Rutgers State Univ, Coll Pharm, Canc Res Lab, 164 Frelinghuysen Rd, Piscataway, NJ 08854 USA. FU NCI NIH HHS [CA49756]; NIEHS NIH HHS [ES03346] NR 31 TC 18 Z9 18 U1 0 U2 3 PU OXFORD UNIV PRESS PI OXFORD PA GREAT CLARENDON ST, OXFORD OX2 6DP, ENGLAND SN 0143-3334 J9 CARCINOGENESIS JI Carcinogenesis PD JUN PY 2001 VL 22 IS 6 BP 951 EP 955 DI 10.1093/carcin/22.6.951 PG 5 WC Oncology SC Oncology GA 440JE UT WOS:000169171500017 PM 11375904 ER PT J AU Kiss, A Ortiz-Aguayo, R Sharp, R Merlino, G Thorgeirsson, SS Gonzalez, FJ Peters, JM AF Kiss, A Ortiz-Aguayo, R Sharp, R Merlino, G Thorgeirsson, SS Gonzalez, FJ Peters, JM TI Evidence that reduction of hepatocyte growth factor (HGF) is not required for peroxisome proliferator-induced hepatocyte proliferation SO CARCINOGENESIS LA English DT Article ID <4-CHLORO-6-(2,3-XYLIDINO)-2-PYRIMIDINYLTHIO>ACETIC ACID WY-14,643; HEPATOCELLULAR-CARCINOMA CELLS; ACTIVATED RECEPTOR; RAT-LIVER; TRANSGENIC MICE; GENE-EXPRESSION; FATTY-ACIDS; MOUSE-LIVER; ALPHA; HEPATOCARCINOGENESIS AB The mechanisms underlying peroxisome proliferator-induced hepatocarcinogenesis are not understood, Because of the uncertainty of human cancer risk associated with peroxisome proliferators, delineating the mechanisms of carcinogenesis by these agents is of great interest. Alterations in liver growth factors were postulated to contribute to the carcinogenic effect of peroxisome proliferators. Administration of these compounds to rodents results in down-regulation of hepatocyte growth factor (HGF) and supplementing culture medium with HGF is reported to suppress cell proliferation of preneoplastic and neoplastic cells from WY-14,643-treated livers, Combined, these observations suggest that reduced levels of hepatic HGF contribute to the mechanisms underlying peroxisome proliferator-induced hepatocarcinogenesis. To determine if HGF can prevent the effects of peroxisome proliferators in liver, the short-term influence of WY-14,643 in two different lines of HGF transgenic mice was examined, Mice were fed either a control diet or one containing 0.1% WY-14-643 for one week. Hepatomegaly was found in both HGF transgenic mouse lines fed WY-14,643 compared with controls. Additionally, hepatic expression of typical mRNA markers of peroxisome proliferation including those encoding peroxisomal fatty acid metabolizing enzymes and cell cycle control proteins were all significantly elevated in HGF transgenic mice fed WY-14,643 compared with controls. Down-regulation of HGF was found to be dependent on PPAR alpha since lower levels of HGF mRNA and protein were observed in wild-type mice fed WY-14,643 for 1 week and not in similarly treated PPAR alpha -null mice, These results demonstrate that the early increase in hepatic mRNAs associated with peroxisome and cell proliferation induced by WY-14,643 treatment can not be prevented by overexpression of HGF in vivo. C1 Penn State Univ, Dept Vet Sci, Ctr Mol Toxicol, University Pk, PA 16802 USA. NCI, Expt Carcinogenesis Lab, Div Basic Sci, Bethesda, MD 20892 USA. NCI, Lab Metab, Div Basic Sci, Bethesda, MD 20892 USA. NCI, Mol Biol Lab, Div Basic Sci, Bethesda, MD 20892 USA. Semmelweis Univ, Inst Pathol, H-1091 Budapest, Hungary. RP Peters, JM (reprint author), Penn State Univ, Dept Vet Sci, Ctr Mol Toxicol, 226 Fenske Lab, University Pk, PA 16802 USA. EM jmp21@psu.edu RI Peters, Jeffrey/D-8847-2011 NR 42 TC 1 Z9 1 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 JUN PY 2001 VL 22 IS 6 BP 975 EP 979 DI 10.1093/carcin/22.6.975 PG 5 WC Oncology SC Oncology GA 440JE UT WOS:000169171500020 PM 11375907 ER PT J AU Kameda, H Risinger, JI Han, BB Baek, SJ Barrett, JC Glasglow, WC Eling, TE AF Kameda, H Risinger, JI Han, BB Baek, SJ Barrett, JC Glasglow, WC Eling, TE TI Identification of epidermal growth factor receptor-Grb2-associated Binder-1-SHP-2 complex formation and its functional loss during neoplastic cell progression SO CELL GROWTH & DIFFERENTIATION LA English DT Article ID PROTEIN-TYROSINE-PHOSPHATASE; TUMOR-SUPPRESSOR GENE; SH2-CONTAINING PHOSPHOTYROSINE PHOSPHATASE; INSULIN-RECEPTOR SUBSTRATE-1; PLECKSTRIN HOMOLOGY DOMAIN; LINOLEIC-ACID METABOLITE; PHOSPHOLIPASE C-GAMMA; HAMSTER EMBRYO CELLS; GRB2 BINDING-SITE; DOCKING PROTEIN AB The adaptor protein Grb2-associated binder-1 (Gab1) is known to bind to the SHP-2 tyrosine phosphatase on epidermal growth factor (EGF) receptor stimulation, To clarify the roles of these two proteins in EGF receptor (EGFR) signaling and determine their possible alteration during neoplastic cell progression, we studied these proteins in a Syrian hamster embryo (SHE) cell line model of neoplastic progression. Specifically, we used ashestos-transformed SHE fibroblasts: the 10W+8 clone, which is immortal but nontumorigenic; and the 10W2T clone, which is tumorigenic. Gab1 was detected, and the EGF-dependent formation of the EGFR-Gab1-SHP-2 complex was observed in 10W+8 cells. After cloning hamster Gab1 cDNA, exogenous expression of Gab1 significantly enhanced EGF-dependent mitogenic activity in 10W+8 cells, On the other hand, Gab1 was not detected in 10W2T cells, and the EGF-dependent association of SHP-2 with EGFR was also absent. Exogenous Gab1 expression in transfected 10W2T cells restored the EGF-dependent association of SHP-2 with EGFR, although it only showed a marginal effect on EGF-dependent mitogenic activity. Thus, Gab1 plays a pivotal role in the EGFR signaling pathway via the formation of the EGFR-Gab1-SHP-2 complex, and alteration in the expression and function of Gab1 is implicated in the neoplastic progression of SHE cells. C1 NIEHS, Mol Carcinogenesis Lab, NIH, Res Triangle Pk, NC 27709 USA. Mercer Univ, Sch Med, Div Basic Med Sci, Macon, GA 31207 USA. RP Eling, TE (reprint author), NIEHS, Mol Carcinogenesis Lab, NIH, POB 12233, Res Triangle Pk, NC 27709 USA. NR 76 TC 8 Z9 9 U1 0 U2 0 PU AMER ASSOC CANCER RESEARCH PI BIRMINGHAM PA PO BOX 11806, BIRMINGHAM, AL 35202 USA SN 1044-9523 J9 CELL GROWTH DIFFER JI Cell Growth Differ. PD JUN PY 2001 VL 12 IS 6 BP 307 EP 318 PG 12 WC Cell Biology SC Cell Biology GA 447ZE UT WOS:000169601700004 PM 11432805 ER PT J AU Litman, T Druley, TE Stein, WD Bates, SE AF Litman, T Druley, TE Stein, WD Bates, SE TI From MDR to MXR: new understanding of multidrug resistance systems, their properties and clinical significance SO CELLULAR AND MOLECULAR LIFE SCIENCES LA English DT Review DE multidrug resistance; P-glycoprotein (MDR1); multidrug-resistance-associated protein (MRP); mitoxantrone resistance protein (MXR, BCRP, ABCP, ABCG2); ABC transporter; ATPase ID HUMAN P-GLYCOPROTEIN; ORGANIC ANION TRANSPORTER; EHRLICH ASCITES TUMOR; HAMSTER OVARY CELLS; PROTEIN MRP GENE; ACUTE MYELOID-LEUKEMIA; ABC HALF-TRANSPORTER; DECREASED DRUG ACCUMULATION; NUCLEOTIDE-BINDING DOMAINS; EMBRYONIC KIDNEY 293-CELLS AB The ATP binding cassette (ABC) superfamily of membrane transporters is one of the largest protein classes known, and counts numerous proteins involved in the trafficking of biological molecules across cell membranes. The first known human ABC transporter was P-glycoprotein (P-gp), which confers multidrug resistance (MDR) to anticancer drugs. In recent years, we have obtained an increased understanding of the mechanism of action of P-gp as its ATPase activity, substrate specificity and pharmacokinetic interactions have been investigated. This review focuses on the functional characterization of P-gp, as well as other ABC transporters involved in MDR: the family of multidrug-resistance-associated proteins (MRP1-7), and the recently discovered ABC half-transporter MXR (also known as BCRP, ABCP and ABCG2). We describe recent progress in the analysis of protein structure-function relationships, and consider the conceptual problem of defining and identifying substrates and inhibitors of MDR. An in-depth discussion follows of how coupling of nucleotide hydrolysis to substrate transport takes place, and we propose a scheme for the mechanism of P-gp function. Finally, the clinical correlations, both for reversal of MDR in cancer and for drug delivery, are discussed. C1 Univ Copenhagen, Panum Inst 12633, Dept Med Physiol, DK-2200 Copenhagen N, Denmark. Univ Illinois, Dept Mol Genet, Chicago, IL 60607 USA. Hebrew Univ Jerusalem, Inst Life Sci, Dept Biol Chem, Jerusalem, Israel. NCI, Med Branch, NIH, Bethesda, MD 20892 USA. RP Litman, T (reprint author), Univ Copenhagen, Panum Inst 12633, Dept Med Physiol, Blegdamsvej 3, DK-2200 Copenhagen N, Denmark. NR 233 TC 510 Z9 528 U1 9 U2 53 PU BIRKHAUSER VERLAG AG PI BASEL PA VIADUKSTRASSE 40-44, PO BOX 133, CH-4010 BASEL, SWITZERLAND SN 1420-682X J9 CELL MOL LIFE SCI JI Cell. Mol. Life Sci. PD JUN PY 2001 VL 58 IS 7 BP 931 EP 959 DI 10.1007/PL00000912 PG 29 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 455CR UT WOS:000170010400007 PM 11497241 ER PT J AU Yang, D Chertov, O Oppenheim, JJ AF Yang, D Chertov, O Oppenheim, JJ TI The role of mammalian antimicrobial peptides and proteins in awakening of innate host defenses and adaptive immunity SO CELLULAR AND MOLECULAR LIFE SCIENCES LA English DT Review DE defensin; chemokine; receptor; cathelicidin; complement; cathepsin G; immunity; antimicrobial ID HUMAN NEUTROPHIL DEFENSINS; HUMAN POLYMORPHONUCLEAR LEUKOCYTES; RICH ANTIBACTERIAL PEPTIDE; TRACHEAL EPITHELIAL-CELLS; IMMATURE DENDRITIC CELLS; VITAMIN-A-DEFICIENCY; CATHEPSIN-G; BETA-DEFENSIN; HUMAN BETA-DEFENSIN-1; ALPHA-DEFENSINS AB Since we live in a dirty environment, we have developed many host defenses to contend with microorganisms. The epithelial lining of our skin, gastrointestinal tract and bronchial tree produces a number of antibacterial peptides, and our phagocytic neutrophils rapidly ingest and enzymatically degrade invading organisms, as well as produce peptides and enzymes with antimicrobial activities. Some of these antimicrobial moieties also appear to alert host cells involved in both innate host defense and adaptive immune responses. The epithelial cells are a source of constitutively produced beta defensin (HBD1) and proinflammatory cytokine-inducible beta defensins (HBD2 and -3) and cathelicidin (LL37). The neutrophils-derived antimicrobial peptides are released on demand from their cytoplasmic granules. They include the enzymes cathepsin G and chymase, azurocidin, alpha defensins and cathelicidin. In contrast, C5a and C3b are produced by activation of the serum complement cascade. The antimicrobial moieties direct the migration and activate target cells by interacting with selected G-protein-coupled seven-transmembrane receptors (GPCRs) on cell surfaces. The beta defensins interact with the CCR6 chemokine GPCRs, whereas cathelicidins interact with the low-affinity FPRL-1 receptors. The neutrophil-derived cathepsin G acts on the high-affinity FMLP receptor (GPCR) known as FPR, while the receptors for chymase and azurocidin have not been identified as yet. The serum-derived C5a uses a GPCR known as C5aR to mediate its chemotactic and cell-activating effects. Consequently, all these ligand-receptor interactions in addition to mediating chemotaxis also activate receptor-expressing cells to produce other mediators of inflammation. C1 NCI, Mol Immunoregulat Lab, Frederick, MD 21702 USA. SAIC, Intramural Res Support Program, Frederick, MD 21702 USA. RP Oppenheim, JJ (reprint author), NCI, Mol Immunoregulat Lab, Bldg 560,Room 21-89A, Frederick, MD 21702 USA. NR 150 TC 168 Z9 186 U1 0 U2 20 PU BIRKHAUSER VERLAG AG PI BASEL PA VIADUKSTRASSE 40-44, PO BOX 133, CH-4010 BASEL, SWITZERLAND SN 1420-682X J9 CELL MOL LIFE SCI JI Cell. Mol. Life Sci. PD JUN PY 2001 VL 58 IS 7 BP 978 EP 989 DI 10.1007/PL00000914 PG 12 WC Biochemistry & Molecular Biology; Cell Biology SC Biochemistry & Molecular Biology; Cell Biology GA 455CR UT WOS:000170010400009 PM 11497243 ER PT J AU Kroth, H Yagi, H Sayer, JM Kumar, S Jerina, DM AF Kroth, H Yagi, H Sayer, JM Kumar, S Jerina, DM TI O-6-allyl protected deoxyguanosine adducts of polycyclic aromatic hydrocarbons as building blocks for the synthesis of oligonucleotides SO CHEMICAL RESEARCH IN TOXICOLOGY LA English DT Article ID BENZOPYRENE DIOL EPOXIDE; SITE-SPECIFIC INCORPORATION; CARCINOGEN-DNA ADDUCTS; DG MISMATCH OPPOSITE; N-RAS CODON-61; DEOXYADENOSINE ADDUCTS; SOLUTION CONFORMATION; 7,8-DIOL 9,10-EPOXIDES; DEPROTECTIVE METHOD; NUCLEOSIDE ADDUCTS AB We describe a synthetic strategy for the preparation of oligonucleotides using N-2-alkylated and O-6-allyl protected deoxyguanosine phosphoramidite building blocks derived from cis- and trans-opened (+/-)-7 beta ,8 alpha -dihydroxy-9 alpha, 10 alpha -epoxy-7,8,9, 10-tetrahydrobenzo[a]pyrene and (+/- )7 beta ,8 alpha -dihydroxy-9 beta, 10 beta -epoxy-7,8,9, 10-tetrahydrobenzo [a]pyrene and from trans-opened (+/-)-3 alpha ,4 beta -dihydroxy- 1 alpha ,2 alpha -epoxy-1,2,3,4-tetrahydrobenzo [c] phenanthrene. The appropriately blocked phosphoramidite building blocks were obtained as mixtures of the cis- and trans-opened diol epoxide adducts upon initial reaction of the diol epoxides with O-6-allyl-3',5'-di-O-(tert-butyldimethylsilyl)-2'-deoxyguanosine. Key to the present approach is the removal of the O-6-allyl protecting group utilizing a palladium catalyst prior to release of the constructed oligonucleotide with ammonia from the solid support. The methodology described enables a very convenient access to oligonucleotides containing cis- and trans-N-2-deoxyguanosine adducts of polycyclic aromatic hydrocarbons in different sequence contexts. C1 NIDDKD, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. SUNY Coll Buffalo, Great Lakes Ctr, Buffalo, NY 14222 USA. RP Jerina, DM (reprint author), NIDDKD, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. NR 53 TC 28 Z9 28 U1 0 U2 4 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0893-228X J9 CHEM RES TOXICOL JI Chem. Res. Toxicol. PD JUN PY 2001 VL 14 IS 6 BP 708 EP 719 DI 10.1021/tx0002637 PG 12 WC Chemistry, Medicinal; Chemistry, Multidisciplinary; Toxicology SC Pharmacology & Pharmacy; Chemistry; Toxicology GA 445TK UT WOS:000169474500010 PM 11409942 ER PT J AU Ponten, I Kroth, H Sayer, JM Dipple, A Jerina, DM AF Ponten, I Kroth, H Sayer, JM Dipple, A Jerina, DM TI Differences between the mutational consequences of replication of cis- and trans-opened benzo[a]pyrene 7,8-diol 9,10-epoxide-deoxyguanosine adducts in M13mp7L2 constructs SO CHEMICAL RESEARCH IN TOXICOLOGY LA English DT Article ID (+)-ANTI-BENZOPYRENE DIOL EPOXIDE; N-RAS CODON-61; TEMPLATE-PRIMER JUNCTION; SITE-SPECIFIC MUTATION; ESCHERICHIA-COLI; SEQUENCE CONTEXT; DNA DUPLEX; SOLUTION CONFORMATION; MAMMALIAN-CELLS; MODIFIED DEOXYGUANOSINE AB The four adducts at N-2 Of deoxyguanosine derived from cis-opening at C-10 of four optically active isomers of 7,8-dihydroxy-9, 10-epoxy-7,8,9,10-tetrahydrobenzo [a]pyrene were incorporated into 5'-TTCGAATCCTTCCCCC [context III(G)] and 5'-GGGGTTCCCGAGCGGC [context IV(G)I at the underlined site. The mutagenic consequences of these lesions in each of the two sequence contexts were examined after ligation of the modified oligonucleotides into single-stranded M13mp7L2 and replication of the vector in SOS-induced Escherichia coli. Total frequencies of base substitution mutations ranged between 14 and 48%. The mutation frequencies were generally higher in context IV(G) than in context III(G), and consisted mainly of G -->T followed by G -->C base substitutions. A substantial number of deletions or insertions of one guanine was also found for all adducts in context IV(G), where the adduct is located at the S'-end of a run of five guanines. The overall frequencies of base substitution mutations induced by cis-opened adducts were substantially higher than those observed with the trans-opened dGuo adducts in the same sequences [Page et al. (1998) Biochemistry 37, 9127-9137]. Although G-T base substitutions predominated for both the cis- and trans-opened adducts, the cis-opened dGuo adducts generally resulted in a higher proportion of G -->C [particularly in context III(G)] relative to G -->A, whereas the opposite was true for the trans-opened dGuo adducts. The present results along with previous data indicate that mutagenicity is highly dependent on a combination of sequence context and adduct stereochemistry. C1 NCI, Chem Carcinogenesis Lab, Frederick, MD 21702 USA. NIDDKD, Bioorgan Chem Lab, NIH, Bethesda, MD 20892 USA. RP Ponten, I (reprint author), NCI, Chem Carcinogenesis Lab, Frederick, MD 21702 USA. NR 45 TC 15 Z9 15 U1 0 U2 0 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0893-228X J9 CHEM RES TOXICOL JI Chem. Res. Toxicol. PD JUN PY 2001 VL 14 IS 6 BP 720 EP 726 DI 10.1021/tx0002684 PG 7 WC Chemistry, Medicinal; Chemistry, Multidisciplinary; Toxicology SC Pharmacology & Pharmacy; Chemistry; Toxicology GA 445TK UT WOS:000169474500011 PM 11409943 ER PT J AU Melnick, RL Sills, RC AF Melnick, RL Sills, RC TI Comparative carcinogenicity of 1,3-butadiene, isoprene, and chloroprene in rats and mice SO CHEMICO-BIOLOGICAL INTERACTIONS LA English DT Article; Proceedings Paper CT 38th European Congress of Toxicology CY SEP 12-14, 2001 CL LONDON, ENGLAND DE butadiene; carcinogenicity; chloroprene; dose-response; ED10; isoprene ID MULTIPLE ORGAN CARCINOGENICITY; INHALATION EXPOSURE; WORKERS; BUTADIENE; TOXICITY AB 1,3-Butadiene, isoprene (2-methyl-1,3-butadiene), and chloroprene (2-chloro-1,3-butadiene) are high-production-volume chemicals used mainly in the manufacture of synthetic rubber. Inhalation studies have demonstrated multiple organ tumorigenic effects with each of these chemicals in mice and rats. Sites of tumor induction by these epoxide-forming chemicals were compared to each other and to ethylene oxide, a chemical classified by the National Toxicology Program (NTP) and by the International Agency for Research on Cancer (IARC) as carcinogenic to humans. For this group of chemicals, there are substantial species differences in sites of neoplasia; neoplasia of the mammary gland is the only common tumorigenic effect in rats and mice. Within each species, there are several common sites of tumor induction; these include the hematopoietic system, circulatory system, lung, liver, forestomach, Harderian gland, and mammary gland in mice, and the mammary gland and possibly the brain, thyroid, testis, and kidney in rats. For studies in which individual animal data were available, mortality-adjusted tumor rates were calculated, and estimates were made of the shape of the exposure-response curves and ED10 values (i.e. exposure concentrations associated with an excess risk of 10% at each tumor site). Most tumorigenic effects reported here were consistent with linear or supralinear models. For chloroprene and butadiene, the most potent response was for the induction of lung neoplasms in female mice, with ED10 values of 0.3 ppm. Based on animal cancer data, isoprene and chloroprene are listed in the NTP's Report on Carcinogens (RoC) as reasonably anticipated to be a human carcinogen. Butadiene is listed in the RoC as known to be a human carcinogen 'based on sufficient evidence of carcinogenicity from studies in humans, including epidemiological and mechanistic information', with support from experimental studies in laboratory animals. Epidemiology data for isoprene and chloroprene are not considered adequate to evaluate the potential carcinogenicity of these agents in humans. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved. C1 NIEHS, Res Triangle Pk, NC 27709 USA. RP Melnick, RL (reprint author), NIEHS, POB 12233, Res Triangle Pk, NC 27709 USA. NR 27 TC 36 Z9 37 U1 1 U2 4 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 JUN 1 PY 2001 VL 135 SI SI BP 27 EP 42 DI 10.1016/S0009-2797(01)00213-7 PG 16 WC Biochemistry & Molecular Biology; Pharmacology & Pharmacy; Toxicology SC Biochemistry & Molecular Biology; Pharmacology & Pharmacy; Toxicology GA 470PU UT WOS:000170881000003 PM 11397379 ER PT J AU Kohn, MC Melnick, RL AF Kohn, MC Melnick, RL TI Physiological modeling of butadiene disposition in mice and rats SO CHEMICO-BIOLOGICAL INTERACTIONS LA English DT Article; Proceedings Paper CT 38th European Congress of Toxicology CY SEP 12-14, 2001 CL LONDON, ENGLAND DE butadiene; enzymatic channeling; metabolic mechanisms; PBPK modeling ID SPRAGUE-DAWLEY RATS; LUNG IN-VITRO; INHALATION PHARMACOKINETICS; SPECIES-DIFFERENCES; GLUTATHIONE CONJUGATION; BLOOD-CONCENTRATIONS; TOXICOKINETIC MODEL; EPOXIDE HYDRASE; MOUSE-LIVER; GAS UPTAKE AB The earliest physiological models of 1,3-butadiene disposition reproduced uptake of the gas from closed chambers but over-predicted steady-state circulating concentrations of the mutagenic intermediates 1,2-epoxybut-3-ene and 1,2:3,4-diepoxybutane. A preliminary model based on the observation of a transient complex between cytochrome P450 and microsomal epoxide hydrolase on the endoplasmic reticulum membrane reproduced the blood epoxide concentrations as well as the chamber uptake data. This model was enhanced by the addition of equations for the production and detoxication of 3,4-epoxybutane-1,2-diol in the liver, lungs, and kidneys. The model includes flow-restricted delivery of butadiene and its metabolites to compartments for lungs, liver, fat, kidneys, gastrointestinal tract, other rapidly perfused tissues, and other slowly perfused tissues. Blood was distributed among compartments for arterial, venous, and tissue capillary spaces. Channeling of the three bound epoxides to epoxide hydrolase and their release from the endoplasmic reticulum are competing processes in this model. Parameters were estimated to fit data for chamber uptake of butadiene and epoxybutene, steady-state blood concentrations of epoxybutene and diepoxybutane, and the fractions of the inhaled dose of butadiene that appears as various excreted metabolites. The optimal values of the apparent K(m)s of membrane-bound epoxides for epoxide hydrolase were only 5% of the values for the cytosolic substrate, consistent with the observation of a transient complex between epoxide hydrolase and the cytochrome P450 that produces the epoxide. This proximity effect corresponds to the notion that epoxides produced in situ have privileged access to epoxide hydrolase. The model also predicts considerable accumulation of epoxybutanediol, in agreement with the observation that most of the DNA adducts in animals exposed to butadiene arise from this metabolite. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved. C1 NIEHS, Lab Computat Biol & Risk Anal, Res Triangle Pk, NC 27709 USA. RP Kohn, MC (reprint author), NIEHS, Lab Computat Biol & Risk Anal, POB 12233,Mail Drop A3-06, Res Triangle Pk, NC 27709 USA. NR 49 TC 12 Z9 13 U1 0 U2 0 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 JUN 1 PY 2001 VL 135 SI SI BP 285 EP 301 DI 10.1016/S0009-2797(01)00176-4 PG 17 WC Biochemistry & Molecular Biology; Pharmacology & Pharmacy; Toxicology SC Biochemistry & Molecular Biology; Pharmacology & Pharmacy; Toxicology GA 470PU UT WOS:000170881000021 PM 11397397 ER PT J AU Sills, RC Hong, HL Boorman, GA Devereux, TR Melnick, RL AF Sills, RC Hong, HL Boorman, GA Devereux, TR Melnick, RL TI Point mutations of K-ras and H-ras genes in forestomach neoplasms from control B6C3F1 mice and following exposure to 1,3-butadiene, isoprene or chloroprene for up to 2-years SO CHEMICO-BIOLOGICAL INTERACTIONS LA English DT Article; Proceedings Paper CT 38th European Congress of Toxicology CY SEP 12-14, 2001 CL LONDON, ENGLAND DE mutations; ras oncogene; forestomach neoplasms; 1,3-butadiene; isoprene; chloroprene; B6C3F1 mouse; chemical carcinogenesis; inhalation studies ID MULTIPLE ORGAN CARCINOGENICITY; SQUAMOUS-CELL CARCINOMAS; METHYL-N-NITROSOUREA; LUNG-TUMORS; PROTOONCOGENE ACTIVATION; INHALATION EXPOSURE; MOUSE FORESTOMACH; DNA-POLYMERASE; RATS; PROLIFERATION AB 1,3 Butadiene (BD), isoprene (IP) and chloroprene (CP) are structural analogs. There were significantly increased incidences of forestomach neoplasms in J36C3F1 mice exposed to BD, IP or CP by inhalation for up to 2-years. The present study was designed to characterize genetic alterations in K- and H-ras proto-oncogenes in a total of 52 spontaneous and chemically induced forestomach neoplasms. ras mutations were identified by restriction fragment length polymorphism, single strand conformational polymorphism analysis, and cycle sequencing of PCR-amplified DNA isolated from paraffin-embedded forestomach neoplasms. A higher frequency of K- and H-ras mutations was identified in BD-, IP- and CP-induced forestomach neoplasms (83, 70 and 57%, respectively, or combined 31/41, 76%) when compared to spontaneous forestomach neoplasms (4/11. 36%). Also a high frequency of H-ras codon 61 CAA --> CTA transversions (10/41, 24%) was detected in chemically induced forestomach neoplasms, but none were present in the spontaneous forestomach neoplasms examined. Furthermore, an increased frequency (treated 13/41, 32% versus untreated 1/11, 9%) of GGC --> CGC transversion at K-ras codon 13 was seen in BD-, and IP-induced forestomach neoplasms, similar to the predominant K-ras mutation pattern observed in BD-induced mouse lung neoplasms. These data suggest that the epoxide intermediates of the structurally related chemicals (BD, IP, and CP) may cause DNA damage in K-ras and H-ras proto-oncogenes of B6C3F1 mice following inhalation exposure and that mutational activation of these genes may be critical events in the pathogenesis of forestomach neoplasms induced in the B6C3F1 mouse. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved. C1 NIEHS, Environm Toxicol Program, Lab Expt Pathol, Res Triangle Pk, NC 27709 USA. NIEHS, Environm Carcinogenesis Program, Res Triangle Pk, NC 27709 USA. RP Sills, RC (reprint author), NIEHS, Environm Toxicol Program, Lab Expt Pathol, POB 12233, Res Triangle Pk, NC 27709 USA. NR 45 TC 11 Z9 12 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 JUN 1 PY 2001 VL 135 SI SI BP 373 EP 386 DI 10.1016/S0009-2797(01)00179-X PG 14 WC Biochemistry & Molecular Biology; Pharmacology & Pharmacy; Toxicology SC Biochemistry & Molecular Biology; Pharmacology & Pharmacy; Toxicology GA 470PU UT WOS:000170881000026 PM 11397402 ER PT J AU Hayes, RB Zhang, L Swenberg, JA Yin, SN Xi, L Wiencke, J Bechtold, WE Yao, M Rothman, N Haas, R O'Neill, JP Wiemels, J Dosemeci, M Li, G Smith, MT AF Hayes, RB Zhang, L Swenberg, JA Yin, SN Xi, L Wiencke, J Bechtold, WE Yao, M Rothman, N Haas, R O'Neill, JP Wiemels, J Dosemeci, M Li, G Smith, MT TI Markers for carcinogenicity among butadiene-polymer workers in China SO CHEMICO-BIOLOGICAL INTERACTIONS LA English DT Article; Proceedings Paper CT 38th European Congress of Toxicology CY SEP 12-14, 2001 CL LONDON, ENGLAND DE butadiene; carcinogenesis; China ID SISTER-CHROMATID EXCHANGES; SYNTHETIC RUBBER WORKERS; OCCUPATIONAL-EXPOSURE; CHROMOSOMAL-ABERRATIONS; MONOMER PRODUCTION; 1,3-BUTADIENE; LYMPHOCYTES; DIEPOXYBUTANE; SENSITIVITY; FREQUENCY AB We examined a spectrum of genotoxic and other outcomes in 41 butadiene-polymer production workers and 38 nonexposed controls, in China, to explore the role of butadiene in human carcinogenesis. Among butadiene-exposed workers, median air exposure was 2 ppm (6-h TWA), due largely to intermittent high-level exposures. Compared to unexposed subjects. butadiene-exposed workers had greater levels of hemoglobin N-(2,3,4-trihydroxybutyl)valine (THBVal) adducts (P<0.0001), and adduct levels tended to correlate, among butadiene-ex posed workers, with air measures (P=0.03). Butadiene-exposed workers did not differ., however, from unexposed workers with respect to frequency of uninduced or diepoxybutane-induced sister chromatid exchanges, aneuploidy as measured by fluorescence in situ hybridization of chromosomes 1, 7, 8 and 12, glycophorin A variants or lymphocyte hprt somatic mutation. Also among the exposed, greater THBVal levels were not associated with increases in uninduced sister chromatid exchanges, aneuploidy, glycophorin A, or hprt mutations. Butadiene-exposed workers had greater lymphocyte (P=0.002) and platelet counts (P=0.07) and lymphocytes as a percent of white blood cells were moderately correlated with greater THBVal levels (Spearman's =0.32, P=0.07). Among butadiene-exposed workers. several serum cytokines correlated with THBVal adduct levels. Overall, the study demonstrated exposure to butadiene in these workers, by a variety of short-term and long-term measures, but did not show specific genotoxic effects, at the chromosomal or gene levels, related to that exposure. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved. C1 NCI, Occupat Epidemiol Branch, Bethesda, MD 20892 USA. Univ Calif Berkeley, Sch Publ Hlth, Berkeley, CA 94720 USA. Univ N Carolina, Sch Publ Hlth & Med, Chapel Hill, NC USA. Chinese Acad Prevent Med, Beijing, Peoples R China. Univ Calif San Francisco, Sch Med, San Francisco, CA USA. Inhalat Toxicol Res Inst, Albuquerque, NM 87185 USA. Calif State Dept Hlth, Berkeley, CA USA. Univ Vermont, Genet Lab, Burlington, VT USA. RP Hayes, RB (reprint author), NCI, Occupat Epidemiol Branch, EPS 8114, Bethesda, MD 20892 USA. EM hayesr@mail.nih.gov NR 24 TC 18 Z9 20 U1 0 U2 5 PU ELSEVIER IRELAND LTD PI CLARE PA ELSEVIER HOUSE, BROOKVALE PLAZA, EAST PARK SHANNON, CO, CLARE, 00000, IRELAND SN 0009-2797 J9 CHEM-BIOL INTERACT JI Chem.-Biol. Interact. PD JUN 1 PY 2001 VL 135 SI SI BP 455 EP 464 DI 10.1016/S0009-2797(01)00182-X PG 10 WC Biochemistry & Molecular Biology; Pharmacology & Pharmacy; Toxicology SC Biochemistry & Molecular Biology; Pharmacology & Pharmacy; Toxicology GA 470PU UT WOS:000170881000030 PM 11397406 ER PT J AU Strable, E Bulte, JWM Moskowitz, B Vivekanandan, K Allen, M Douglas, T AF Strable, E Bulte, JWM Moskowitz, B Vivekanandan, K Allen, M Douglas, T TI Synthesis and characterization of soluble iron oxide-dendrimer composites SO CHEMISTRY OF MATERIALS LA English DT Article ID PERSUBSTITUTED POLY(AMIDOAMINE) DENDRIMERS; MR CONTRAST AGENT; SILVER NANOPARTICLES; AQUEOUS-SOLUTION; PROTEIN CAGES; NANOCOMPOSITES; NANOCLUSTERS; GOLD; MAGNETOLIPOSOMES; MAGNETOFERRITIN AB Carboxylated poly(amidoamine) PAMAM dendrimers (generation 4.5) have been utilized for the synthesis and stabilization of ferrimagnetic iron oxide nanoparticles. Oxidation of Fe(II) at slightly elevated pH and temperature results in the formation of highly soluble nanocomposites of iron oxides and dendrimer, which are stable under a wide range of temperatures and pH's. Size exclusion chromatography indicates aggregates in the 20-30-nm size regime, consistent with the oligomeric nature of the composite material. Transmission electron microscopy reveals small assemblies of mineral cores with electron diffraction and high-resolution TEM data, indicative of the mineral maghemite. SQUID magnetometry demonstrated that this crystalline composition exhibited superparamagnetism at room temperature. NMR relaxation studies of solvent (water) protons revealed unusually high T1 and T2 relaxivities, which make these materials excellent candidates as contrast agents for MR imaging. C1 Temple Univ, Dept Chem, Philadelphia, PA 19122 USA. NIH, Lab Diagnost Radiol Res, Bethesda, MD 20892 USA. Univ Minnesota, Dept Geol & Geophys, Inst Rock Magnetism, Minneapolis, MN 55455 USA. RP Douglas, T (reprint author), Temple Univ, Dept Chem, Beury Hall, Philadelphia, PA 19122 USA. EM tdouglas@nimbus.temple.edu RI Bulte, Jeff/A-3240-2008; Douglas, Trevor/F-2748-2011 OI Bulte, Jeff/0000-0003-1202-1610; NR 39 TC 135 Z9 138 U1 5 U2 44 PU AMER CHEMICAL SOC PI WASHINGTON PA 1155 16TH ST, NW, WASHINGTON, DC 20036 USA SN 0897-4756 J9 CHEM MATER JI Chem. Mat. PD JUN PY 2001 VL 13 IS 6 BP 2201 EP 2209 DI 10.1021/cm010125i PG 9 WC Chemistry, Physical; Materials Science, Multidisciplinary SC Chemistry; Materials Science GA 445TD UT WOS:000169473900039 ER PT J AU Escalante, P Graviss, EA Griffith, DE Musser, JM Awe, RJ AF Escalante, P Graviss, EA Griffith, DE Musser, JM Awe, RJ TI Treatment of isoniazid-resistant tuberculosis in southeastern Texas SO CHEST LA English DT Article DE drug resistance; isoniazid; isoniazid resistance; relapse; treatment; tuberculosis ID PULMONARY TUBERCULOSIS; DRUG RESISTANCE AB Background: Isoniazid-resistant tuberculosis (INHr-TB) can be treated successfully with several treatment regimens, However, the optimal regimen and duration are unclear, Study objective: To analyze the efficacy of treatment regimens used for INHr-TB in the southeastern Texas region. Design: Retrospective cohort study, Setting: Health-care facilities reporting tuberculosis (TB) patients in the Houston and Tyler areas. Subjects: All patients reported to have INHr-TB from 1991 to 1998, Exclusion criteria included poor compliance, additional first-line drug-resistance (except aminoglycosides), and death before completion of 1 month of treatment. Measurements and results: Main treatment outcomes were treatment failure, relapse, and TB-related death. Fifty-three of 83 patients were included in the study; aminoglycoside resistance coexisted in 37.5% of isolates, Seven types of treatment regimens were identified. Eighteen patients (34%) received rifampin, pyrazinamide, and ethambutol thrice weekly for 9 months. Four patients (7.5%) had a total effective treatment duration of < 9 months. Thirty patients (56.6%) and 16 patients (30.2%) received thrice-daily and daily treatment regimens, respectively, Forty-nine patients achieved sputum conversion, Treatment failure and death occurred in one patient (1.9%), Three patients (5.7%) experienced relapses. There was a significant difference in total effective treatment time between patients with and without relapses (8.3 +/- 1.1 months vs 11.1 +/- 2.1 months; p < 0.02), Twice-weekly treatment regimens were associated with relapse (p = 0.05), Conclusions: Several treatment regimens were prescribed for INHr-TB in southeastern Texas. INHr-TB treatment durations were > 7 months, and treatment regimen efficacy was adequate. Twice-weekly treatment was associated with relapse, whereas thrice-weekly and daily treatments performed similarly, A prospective study with different treatment durations is needed to determine the optimal treatment regimen for patients with INHr-TB. C1 Baylor Coll Med, Inst Study Human Bacterial Pathogenesis, Sect Pulm & Crit Care, Houston, TX 77030 USA. Baylor Coll Med, Inst Study Human Bacterial Pathogenesis, Infect Dis Sect, Houston, TX 77030 USA. Univ Texas, Ctr Hlth, Ctr Pulm & Infect Dis Control, Tyler, TX 75710 USA. NIAID, Rocky Mt Labs, Lab Human Bacterial Pathogenesis, NIH, Hamilton, MT 59840 USA. RP Escalante, P (reprint author), Univ So Calif, Los Angeles Cty Med Ctr, Dept Med, Keck Sch Med,Div Pulm & Crit Care, 1200 N State St,GNH 11900, Los Angeles, CA 90033 USA. FU NIDA NIH HHS [DA-09238] NR 17 TC 12 Z9 12 U1 0 U2 0 PU AMER COLL CHEST PHYSICIANS PI NORTHBROOK PA 3300 DUNDEE ROAD, NORTHBROOK, IL 60062-2348 USA SN 0012-3692 J9 CHEST JI Chest PD JUN PY 2001 VL 119 IS 6 BP 1730 EP 1736 DI 10.1378/chest.119.6.1730 PG 7 WC Critical Care Medicine; Respiratory System SC General & Internal Medicine; Respiratory System GA 441NB UT WOS:000169235500020 PM 11399698 ER PT J AU Wang, FS Jin, L Liu, MX Hong, WG Shi, H Lei, ZY Wang, JM Du, QY Hou, J Zhang, B Jiang, JD Wang, Y Li, YQ Xu, AL Huang, HL Liu, FJ AF Wang, FS Jin, L Liu, MX Hong, WG Shi, H Lei, ZY Wang, JM Du, QY Hou, J Zhang, B Jiang, JD Wang, Y Li, YQ Xu, AL Huang, HL Liu, FJ TI Polymorphisms of chemokine receptors and its ligand alleles influencing genetic susceptibity to HIV-1 infection in eight ethnic groups in Chinese mainland SO CHINESE SCIENCE BULLETIN LA English DT Article DE HIV-1; coreceptors; polymorphism; allelic frequency; mutation ID DISEASE PROGRESSION; CCR5 GENE; PATHOGENESIS; VARIANTS AB polymorphisms of HIV-1 resistant genes in indigenous Chinese populations. The aim of this study is to identify the allelic frequencies of the chemokine and chemokine receptor genes in the Chinese mainland. Genomic DNA samples extracted from whole blood of 2318 subjects were analyzed by using PCR or PCR/restriction fragment length polymorphism (RFLP) assays, and further confirmed by direct DNA sequencing. Higher frequencies of mutant CCR2-641 (19.15%-28.79%) and SDFI-3'A (19.10%-29.86%) alleles were found in subjects of 8 ethnic groups in the Chinese mainland. In contrast, the Delta 32 mutation in CCR5 gene occurs at a very low frequency (0.0016, n=1287) in Han population. A relatively high frequency of CCR5-wt/Delta 32 heterozygotes was observed in Uygurian and Mongolian populations. No Delta 32 mutation allele was detected in Tibetan and other 4 ethnic groups in Yunnan Province. There was no CCR5-m303 mutation in subjects of any ethnic group in the Chinese mainland. Our results suggest that the CCR5-Delta 32 mutation is not a major resistant factor against HIV-1 infection and disease progression in Han, Tibetan and other ethnic groups in Yunnan Province. Whether higher frequencies of CCR2-641 and SDF1-3'A alleles constitute major genetic resistant factors or not remains to be clarified. C1 Beijing Hosp Infect Dis, Beijing Inst Infect Dis, Dept Outpatients, Div Biol Engn, Beijing 100039, Peoples R China. NCI, Mol Immunoregulat Lab, Div Basic Sci, FCRDC, Frederick, MD 21702 USA. CUNY Mt Sinai Sch Med, Div Med Oncol, New York, NY 10029 USA. Zhongshan Univ, Acad Med Sci, Key Lab Mol Biol, Guangzhou 510275, Peoples R China. Kashi Hlth & AntiEpidem Ctr, Kashi 844001, Xinjiang, Peoples R China. RP Wang, FS (reprint author), Beijing Hosp Infect Dis, Beijing Inst Infect Dis, Dept Outpatients, Div Biol Engn, Beijing 100039, Peoples R China. NR 16 TC 1 Z9 3 U1 1 U2 4 PU SCIENCE PRESS PI BEIJING PA 16 DONGHUANGCHENGGEN NORTH ST, BEIJING 100717, PEOPLES R CHINA SN 1001-6538 J9 CHINESE SCI BULL JI Chin. Sci. Bull. PD JUN PY 2001 VL 46 IS 12 BP 1019 EP 1023 PG 5 WC Multidisciplinary Sciences SC Science & Technology - Other Topics GA 458KQ UT WOS:000170193900013 ER PT J AU Vecchione, A Ishii, H Shiao, YH Trapasso, F Rugge, M Tamburrino, JF Murakumo, Y Alder, H Croce, CM Baffa, R AF Vecchione, A Ishii, H Shiao, YH Trapasso, F Rugge, M Tamburrino, JF Murakumo, Y Alder, H Croce, CM Baffa, R TI Fez1/Lzts1 alterations in gastric carcinoma SO CLINICAL CANCER RESEARCH LA English DT Article ID ADENOCARCINOMA; CANCER; HETEROZYGOSITY; EXPRESSION AB Purpose: Loss of heterozygosity (LOH) involving the short arm of chromosome 8 (8p) is a common feature of the malignant progression of human tumors, including gastric cancer, We have cloned and mapped a candidate tumor suppressor gene, FEZ1/LZTS1, to 8p22, Here we have analyzed whether FEZ1/LZTS1 alterations play a role in the development and progression of gastric carcinoma. Experimental Design: We examined Fez1/LZTS1 expression in 8 gastric carcinoma cell lines by Western blot, and in 88 primary gastric carcinomas by immunohistochemistry. Twenty-six of these 88 primary gastric carcinomas were also microdissected and tested for LOH at the FEZ1/LZTS1 locus and for mutation of the FEZI/LZTSI gene. Furthermore, we studied the FEZ1/LZTS1 gene regulation and transcriptional control and the methylation status of the 5' region of the gem in ah 8 gastric carcinoma cell lines. Results: Fez1/Lzts1 protein was barely detectable in all of the gastric cancer cell lines tested and was absent or significantly reduced in 39 of the 88 (44.3%) gastric tarcinomas analyzed by immunohistochemistry,,vith a significant correlation (P < 0.001) to diffuse histotype, DNA allelotyping analysis showed allelic loss in 3 of 17 (18%) and microsatellite instability in 4 of 17 (23.5%) cases informative for D8S261 at the FEZ1/LZTS1 locus. When we compared the presence of LOH with Fez1/Lzts1 expression, we found loss of protein expression in all three of the tumors with allelic imbalance at D8S261, A missense mutation was detected in one case that did not express Fez1/Lzts1, Hypermethylation of the CpG island flanking the Fez1/Lzts1 promoter was evident in six of the eight tell lines examined as well as in the normal control. Conclusions: Our findings support FEZI/LZTSI as a candidate tumor suppressor gene at gp in a subtype of gastric cancer and suggest that its inactivation is attributable to several factors including genomic deletion and methylation. C1 Thomas Jefferson Univ, Jefferson Med Coll, Kimmel Canc Ctr, Philadelphia, PA 19107 USA. NCI, Frederick Canc Res & Dev Ctr, Comparat Carcinogenesis Lab, Frederick, MD 21702 USA. Univ Padua, Dept Pathol, I-35126 Padua, Italy. RP Baffa, R (reprint author), Thomas Jefferson Univ, Jefferson Med Coll, Kimmel Canc Ctr, 1015 Walnut St,Suite 1102A, Philadelphia, PA 19107 USA. RI Rugge, Massimo/K-7525-2016 FU NCI NIH HHS [CA83698-01A1] NR 16 TC 43 Z9 52 U1 0 U2 1 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 JUN PY 2001 VL 7 IS 6 BP 1546 EP 1552 PG 7 WC Oncology SC Oncology GA 442XQ UT WOS:000169310600010 PM 11410489 ER PT J AU Shapiro, GI Supko, JG Patterson, A Lynch, C Lucca, J Zacarola, PF Muzikansky, A Wright, JJ Lynch, TJ Rollins, BJ AF Shapiro, GI Supko, JG Patterson, A Lynch, C Lucca, J Zacarola, PF Muzikansky, A Wright, JJ Lynch, TJ Rollins, BJ TI A phase II trial of the cyclin-dependent kinase inhibitor flavopiridol in patients with previously untreated stage IV non-small cell lung cancer SO CLINICAL CANCER RESEARCH LA English DT Article ID BREAST-CARCINOMA CELLS; NONSMALL CELL; TUMOR SUPPRESSION; DOWN-REGULATION; SOLID TUMORS; APOPTOSIS; INACTIVATION; ARREST; P16(INK4A); PROTEIN AB Purpose: Flavopiridol is a patent cyclin-dependent kinase inhibitor with preclinical activity against non-small cell lung cancer (NSCLC), inhibiting tumor growth in vitro and in vivo by cytostatic and cytotoxic mechanisms. A Phase IT trial was conducted to determine the activity and toxicity of flavopiridol in untreated patients with metastatic NSCLC, Experimental Design: A total of 20 patients were treated with a 72-h continuous infusion of flavopiridol every 14 days at a dose of 50 mg/m(2)/day and a concentration of 0.1-0.2 mg/ml, Dose escalation to 60 mg/m(2)/day was permitted if no significant toxicity occurred. Response was initially assessed after every two infusions; patients treated longer than 8 weeks were then assessed after every four infusions. Plasma levels of flavopiridol were measured daily during the first two infusions to determine steady-state concentrations. Results: This study was designed to evaluate a total of 45 patients in two stages. However, because no objective responses were seen in the first 20 patients, the early-stopping rule was invoked, and patient accrual was halted. In four patients who received eight infusions, progression was documented at 15, 20, 40, and 65 weeks, respectively, The most common toxicities included grade 1 or 2 diarrhea in 11 patients, asthenia in 10 patients, and venous thromboses in 7 patients. The mean +/- SD steady-state concentration of drug during the first infusion was 200 +/- 89.9 nM, sufficient for cytostatic effects in irt vitro models, Conclusions: At the current doses and schedule, flavopiridol does not have cytotoxic activity in NSCLC, although protracted periods of disease stability were observed with an acceptable degree of toxicity. C1 Dana Farber Canc Inst, Lowe Ctr Thorac Oncol, Boston, MA 02115 USA. Massachusetts Gen Hosp, Ctr Canc, Boston, MA 02114 USA. NCI, Div Canc Treatment & Diagnost, Canc Therapy Evaluat Program, Bethesda, MD 20892 USA. RP Shapiro, GI (reprint author), Dana Farber Canc Inst, Lowe Ctr Thorac Oncol, Mayer 422B,44 Binney St, Boston, MA 02115 USA. FU NCI NIH HHS [U01 CA62490-07] NR 65 TC 127 Z9 134 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 JUN PY 2001 VL 7 IS 6 BP 1590 EP 1599 PG 10 WC Oncology SC Oncology GA 442XQ UT WOS:000169310600016 PM 11410495 ER PT J AU Kohn, EC Reed, E Sarosy, GA Minasian, L Bauer, KS Bostick-Bruton, F Kulpa, V Fuse, E Tompkins, A Noone, M Goldspiel, B Pluda, J Figg, WD Liotta, LA AF Kohn, EC Reed, E Sarosy, GA Minasian, L Bauer, KS Bostick-Bruton, F Kulpa, V Fuse, E Tompkins, A Noone, M Goldspiel, B Pluda, J Figg, WD Liotta, LA TI A phase I trial of carboxyamido-triazole and paclitaxel for relapsed solid tumors: Potential efficacy of the combination and demonstration of pharmacokinetic interaction SO CLINICAL CANCER RESEARCH LA English DT Article ID MEDIATED SIGNAL-TRANSDUCTION; REFRACTORY OVARIAN-CANCER; COLONY-STIMULATING FACTOR; CALCIUM INFLUX; CYTOTOXIC THERAPIES; TAXOL; ANGIOGENESIS; CELLS; INHIBITOR; CHEMOTHERAPY AB Purpose: Preclinical and clinical investigation of the combination of the antiangiogenesis/anti-invasion agent carboxyamido-triazole (CAI) administered with the cytotoxic agent paclitaxel (PAX), Experimental Design: Colony-forming assays were used to test the activity of CAI plus PAX on A2780 human ovarian cancer. The sequence of CAI followed by PAX (CAI > Pax) was modeled in nude mice to test for potential additive toxicity, The Phase I clinical dose escalation schema tested p.o. administered CAI in PEG-400 (50-100 mg/m(2)) or micronized CAI (250 mg/m(2)) for 8 days followed by a 3-h infusion of PAX (110-250 mg/m(2)) every 21 days, Patients were assessed for toxicity, pharmacokinetics of CAI and PAX, and disease outcome. Results: In preclinical studies, CAI>Pax was additive in A2780 human ovarian cancer cell lines when CAI (1 or 5 muM) preceded subtherapeutic doses of PAX. CAI did not reverse PAX resistance and collateral resistance to CAI was documented in PAX-resistant cells. CAI > PAX administration had no overt additive toxicity in nude mice. Thirty-nine patients were treated on a dose-escalation Phase I trial using daily oral CAI for 8 days followed by the PAX infusion. Pharmacokinetic analysis revealed that PAX caused an acute increase in circulating CAI concentrations in a dose-dependent fashion. No additive or cumulative toxicity was observed, and grade 3 nonhematological toxicity was rare, Three partial responses and two minor responses were observed. Conclusions: The sequential combination of CAI and PAX is well tolerated, and the activity observed suggests that further study of the combination is warranted. C1 NCI, Med Branch, Bethesda, MD 20892 USA. NCI, Pathol Lab, Bethesda, MD 20892 USA. NCI, Canc Therapy Evaluat Program, Bethesda, MD 20892 USA. NIH, Warren G Magnuson Clin Ctr, Bethesda, MD 20892 USA. RP Kohn, EC (reprint author), 10 Ctr Dr MSC1500, Bethesda, MD 20892 USA. RI Figg Sr, William/M-2411-2016 NR 52 TC 40 Z9 40 U1 1 U2 3 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 JUN PY 2001 VL 7 IS 6 BP 1600 EP 1609 PG 10 WC Oncology SC Oncology GA 442XQ UT WOS:000169310600017 PM 11410496 ER PT J AU Kang, MH Figg, WD Ando, Y Blagosklonny, MV Liewehr, D Fojo, T Bates, SE AF Kang, MH Figg, WD Ando, Y Blagosklonny, MV Liewehr, D Fojo, T Bates, SE TI The P-glycoprotein antagonist PSC 833 increases the plasma concentrations of 6 alpha-hydroxypaclitaxel, a major metabolite of paclitaxel SO CLINICAL CANCER RESEARCH LA English DT Article ID RESISTANCE GENE-PRODUCT; HUMAN LIVER-MICROSOMES; MULTIDRUG-RESISTANCE; CYTOCHROME-P450 3A4; BIOLOGICAL-ACTIVITY; DRUG-INTERACTIONS; TAXOL METABOLISM; CYCLOSPORINE-A; PHASE-II; EXPRESSION AB Purpose: Overexpression of P-glycoprotein (Pgp) is one mechanism of drug resistance in cancer chemotherapy, A Phase I trial was conducted using PSC 833, a Pgp antagonist, in combination with paclitaxel in patients with refractory cancer. The objective of this study was to assess the effect of PSC 833 on the metabolism of paclitaxel and characterize the differences in 6 alpha -hydroxypaclitaxel pharmacokinetics. In addition, we examined the possibility of enhanced cytotoxicity of paclitaxel by the coexistence of 6 alpha -hydroxypaclitaxel. Experimental Design: Patients received paclitaxel 35 mg/m(2)/day by continuous intravenous infusion (CIVI) x 4 days without PSC 833 in cycle I and escalating doses of paclitaxel (13.1, 17.5, or 21.3 mg/m(2)/day CIVI x 4 days) with 5 mg/kg PSC 833 by mouth every 6 h x 7 days in cycle 2. Plasma samples were analyzed for both paclitaxel and its major metabolite with high-performance liquid chromatography methods. Using human liver microsomes, we studied the effect of PSC 833 on the metabolism of paclitaxel, In addition, the in vitro cytotoxicity of 6 alpha -hydroxypaclitaxel alone and in combination with paclitaxel was evaluated, Results,, Twenty-one of 22 patients had a metabolite peak (6 alpha -hydroxypaclitaxel) observed in the chromatogram of plasma samples from cycle 2 when they received paclitaxel in combination with PSC 833, This metabolite was not detectable in plasma obtained during the first cycle when they received paclitaxel without PSC 833, During cycle 2, the mean concentrations of 6 alpha -hydroxypaclitaxel and paclitaxel were 0.10 +/- 0.074 and 0.079 +/- 0.041 mug/ml, respectively. A moderate association was observed between total bilirubin and 6a-hydroxypaclitaxel concentrations (P = 0.015, r 0.52; n = 21), Human liver microsome experiments showed that a PSC 833 concentration as high as 10 phr did not affect the production of 6 alpha -hydroxypaclitaxel. Paclitaxel cytotoxicity in HL60 and K562 human leukemia cells was increased in the presence of noncytotoxic concentrations of 6 alpha -hydroxypaclitaxel. Conclusions: PSC 833 increases the plasma concentration of 6 alpha -hydroxypaclitaxel during paclitaxel therapy, Inhibition of cytochrome P-450 3A4 by PSC 833 may explain this in part, although other mechanisms cannot be excluded. C1 NCI, Med Branch, NIH, Bethesda, MD 20892 USA. NCI, Biostat & Data Management Sect, NIH, Bethesda, MD 20892 USA. NCI, Div Clin Sci, NIH, Bethesda, MD 20892 USA. RP Figg, WD (reprint author), NCI, Med Branch, NIH, Bldg 10,Room 5A01,9000 Rockville Pike, Bethesda, MD 20892 USA. RI Figg Sr, William/M-2411-2016 NR 39 TC 26 Z9 26 U1 1 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 JUN PY 2001 VL 7 IS 6 BP 1610 EP 1617 PG 8 WC Oncology SC Oncology GA 442XQ UT WOS:000169310600018 PM 11410497 ER PT J AU Dudkin, L Dilling, MB Cheshire, PJ Harwood, FC Hollingshead, M Arbuck, SG Travis, R Sausville, EA Houghton, PJ AF Dudkin, L Dilling, MB Cheshire, PJ Harwood, FC Hollingshead, M Arbuck, SG Travis, R Sausville, EA Houghton, PJ TI Biochemical correlates of mTOR inhibition by the rapamycin ester CCI-779 and tumor growth inhibition SO CLINICAL CANCER RESEARCH LA English DT Article ID P70 S6 KINASE; MESSENGER-RNA; PHAS-I; RHABDOMYOSARCOMA CELLS; MAMMALIAN PROTEIN; CANCER-CELLS; (S/T)P SITES; T-CELLS; PHOSPHORYLATION; TRANSLATION AB The rapamycin ester, CCI-779, potently inhibits cell growth in vitro, inhibits tumor growth in vivo, and is currently in Phase I clinical trials. To further understand the relationship between plasma systemic exposure and inhibition of the target Ser/Thr kinase, mTOR/FRAP, two assays have been developed. The first assay involves determination of the 4E suppressor protein (4E-BP1) bound to eukaryotic initiation factor 4E (eIF4E), and the second is direct Western analysis of phosphorylation of residue Thr(70) Of 4E-BP1. Under normal growth conditions in vitro, rapamycin caused rapid association of 4E-BP1 with eIF4E within 1 h in Rh30 and GC(3) human tumor cells, Association was persistent up to 16 h. In mice, administration of rapamycin (5 or 20 mg/kg) caused rapid association of 4E-BP1 with eIF4E within 4 h in both human colon adenocarcinoma GC, and rhabdomyosarcoma Rh30 xenografts, Using phosphospecific antibody against Thr(70) of 4E-BP1, rapid and persistent dephosphorylation within 30 min of exposure to rapamycin was detected in Rh18 rhabdomyosarcoma cells. Evaluation of CCI-779 against Rh18 xenografts showed this tumor to be growth inhibited at daily dose levels of greater than or equal to8.7 mg/kg, Because immunoblotting may be more suitable for assaying tumor biopsy tissue, a "blinded" comparison between the effect of CCI-779 on Thr(70) phosphorylation and growth inhibition of human tumor xenografts was undertaken. Mice were treated daily for 5 days with CCI-779 (20 mg/kg/day) or with drug vehicle, and tumor diameters were measured. Tumors were excised I h after the final administration and frozen, and phaspho Thr(70) was determined by Western blot analysis. The correlation coefficient for decreases in Thr(70) phosphorylation and growth inhibition was high (r(2), 0.99). The results indicate that an assay of decreases in phosphorylation of Thr(70) Of 4E-BP1 may be a useful surrogate for determining the inhibition of mTOR activity in tumor specimens. C1 St Jude Childrens Res Hosp, Dept Mol Pharmacol, Memphis, TN 38105 USA. NCI, Dev Therapeut Program, NIH, Bethesda, MD 20892 USA. NCI, Invest Drug Branch, Canc Therapy Evaluat Program, NIH, Bethesda, MD 20892 USA. NCI, Frederick Canc Res & Dev Ctr, Sci Applicat Int Corp, Frederick, MD 21702 USA. RP Houghton, PJ (reprint author), St Jude Childrens Res Hosp, Dept Mol Pharmacol, 332 N Lauderdale St, Memphis, TN 38105 USA. FU NCI NIH HHS [CA21765, CA23099, CA77776, N01-CO-56000] NR 43 TC 202 Z9 205 U1 0 U2 1 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 JUN PY 2001 VL 7 IS 6 BP 1758 EP 1764 PG 7 WC Oncology SC Oncology GA 442XQ UT WOS:000169310600038 PM 11410517 ER PT J AU Wolf, JS Chen, Z Dong, G Sunwoo, JB Bancroft, CC Capo, DE Yeh, NTY Mukaida, N Van Waes, C AF Wolf, JS Chen, Z Dong, G Sunwoo, JB Bancroft, CC Capo, DE Yeh, NTY Mukaida, N Van Waes, C TI IL (interleukin)-1 alpha promotes nuclear factor-kappa B and AP-1-induced IL-8 expression, cell survival, and proliferation in head and neck squamous cell carcinomas SO CLINICAL CANCER RESEARCH LA English DT Article ID PROINFLAMMATORY CYTOKINE EXPRESSION; TUMOR-NECROSIS-FACTOR; INTERLEUKIN-1 RECEPTOR ANTAGONIST; HOST ENVIRONMENT PROMOTES; CONSTITUTIVE ACTIVATION; TRANSCRIPTION FACTORS; BREAST-CANCER; GROWTH; ALPHA; METASTASIS AB Interleukin 1 alpha (IL-1 alpha) is an important regulatory cytokine, the release of which after an injury can induce activation of transcription factors nuclear factor (NF)kappaB and activator protein (AP-I), which promote expression of genes involved in cell survival, proliferation, and angiogenesis. IL-1 alpha is expressed autonomously by head and neck squamous cell carcinomas (HNSCCs) and a variety of other cancers, raising the possibility that IL-1 alpha may serve as an autocrine factor that stimulates the activation of prosurvival transcription factors and target genes in cancer. In this study, we examined the role of IL-1 alpha in the activation of NF kappaB and AP-I, the expression of proangiogenic cytokine IL-g, and in the survival and proliferation of HNSCC cell lines. HNSCCs were found to secrete and respond to functional IL-1 alpha, in that culture supernatant from a high IL-1 alpha -secreting line, UM-SCC-IIB, could induce secretion of cytokine IL-g by a low IL-1 alpha -secreting Line, UM-SCC-9; and the induction of IL-8 secretion could be blocked by the anti-IL-1 alpha -neutralizing antibody or the IL-1 receptor antagonist (IL-1RA), Furthermore, IL-1 alpha could induce the expression of IL-8 through an autocrine mechanism, in that transfection of UM-SCC-9 cells with a plasmid encoding IL-lat resulted in the increased coexpression of IL-1 alpha. and IL-8; whereas transfection with a plasmid encoding IL-1RA lacking the secretory leader sequence led to the decreased coexpression of HL-1 alpha and IL-g, IL-1 alpha was found to induce coexpression of IL-8 through the activation of NF kappaB and AP-1, in that mutation of the NF kappaB site within the IL-8 promoter abolished autocrine- and recombinant IL-1 alpha -induced IL-8 reporter gene activity, whereas mutation in AP-I partially decreased IL-8 reporter gene activity in UM-SCC-9 cells, Intracellular expression of IL-1RA decreased NF kappaB reporter gene activity, indicating that endogenously expressed IL-1 alpha contributes to constitutive NF kappaB activation in this HNSCC line, Expression of IL-1 alpha affected survival of UM-SCC-9, inasmuch as transfection of cells with plasmid encoding IL-1 alpha or IL-IRA led to the increased or decreased survival of cells cotransfected with a beta -galactosidase reporter gene, respectively. IL-1 alpha was also found to promote the increased growth of UM-SCC-9 cells in vitro, We demonstrate that exogenous and endogenous IL-1 alpha contributes to the transcriptional activation of NF kappaB and AP-I, to the expression of IL-g, and to cell survival and the growth of HNSCC in vitro. C1 Natl Inst Deafness & Other Commun Disorders, Tumor Biol Sect, Head & Neck Surg Branch, NIH, Bethesda, MD 20892 USA. Kanazawa Univ, Canc Res Inst, Div Mol Pharmacol, Kanazawa, Ishikawa 920, Japan. RP Van Waes, C (reprint author), Natl Inst Deafness & Other Commun Disorders, Tumor Biol Sect, Head & Neck Surg Branch, NIH, Bldg 10,Room 5D55,MSC-1419, Bethesda, MD 20892 USA. RI Mukaida, Naofumi/D-7623-2011 OI Mukaida, Naofumi/0000-0002-4193-1851 FU NIDCD NIH HHS [Z01-DC-00016] NR 44 TC 160 Z9 162 U1 0 U2 4 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 JUN PY 2001 VL 7 IS 6 BP 1812 EP 1820 PG 9 WC Oncology SC Oncology GA 442XQ UT WOS:000169310600045 PM 11410524 ER PT J AU Eisenhofer, G AF Eisenhofer, G TI Free or total metanephrines for diagnosis of pheochromocytoma: What is the difference? SO CLINICAL CHEMISTRY LA English DT Editorial Material ID PLASMA METANEPHRINES; METHOXYAMINES; METABOLITES; URINARY C1 NINDS, Clin Neuroradiol Sect, NIH, Bethesda, MD 20892 USA. RP Eisenhofer, G (reprint author), NINDS, Clin Neuroradiol Sect, NIH, Bethesda, MD 20892 USA. NR 15 TC 39 Z9 44 U1 0 U2 0 PU AMER ASSOC CLINICAL CHEMISTRY PI WASHINGTON PA 2101 L STREET NW, SUITE 202, WASHINGTON, DC 20037-1526 USA SN 0009-9147 J9 CLIN CHEM JI Clin. Chem. PD JUN PY 2001 VL 47 IS 6 BP 988 EP 989 PN 1 PG 2 WC Medical Laboratory Technology SC Medical Laboratory Technology GA 437MW UT WOS:000168996300002 PM 11375282 ER PT J AU Hortin, GL Sullivan, P Csako, G AF Hortin, GL Sullivan, P Csako, G TI Relationships among plasma homocysteine, cysteine, and albumin concentrations: Potential utility of assessing the cysteine/homocysteine ratio SO CLINICAL CHEMISTRY LA English DT Letter ID PROTEIN-BOUND HOMOCYSTEINE; COMPONENTS; DISEASE C1 NIH, Warren G Magnuson Clin Ctr, Dept Lab Med, Bethesda, MD 20892 USA. RP Hortin, GL (reprint author), NIH, Warren G Magnuson Clin Ctr, Dept Lab Med, Bldg 10,Room 2C-407, Bethesda, MD 20892 USA. NR 17 TC 10 Z9 11 U1 0 U2 0 PU AMER ASSOC CLINICAL CHEMISTRY PI WASHINGTON PA 2101 L STREET NW, SUITE 202, WASHINGTON, DC 20037-1526 USA SN 0009-9147 J9 CLIN CHEM JI Clin. Chem. PD JUN PY 2001 VL 47 IS 6 BP 1121 EP 1124 PN 1 PG 4 WC Medical Laboratory Technology SC Medical Laboratory Technology GA 437MW UT WOS:000168996300027 PM 11375307 ER PT J AU Fulgenzi, A Wasserman, K Corsi, MM AF Fulgenzi, A Wasserman, K Corsi, MM TI Significance of autoantibodies to oxidatively modified LDL in plasma of children with Down syndrome SO CLINICAL CHEMISTRY LA English DT Letter ID LOW-DENSITY-LIPOPROTEIN; ATHEROSCLEROSIS; ANTIOXIDANTS; EXPRESSION; HYPOTHESIS; ATHEROMA; PROTEIN C1 Univ Milan, Fac Med, Inst Gen Pathol, I-20133 Milan, Italy. NCI, Frederick Canc Res & Dev Ctr, Lab Expt Immunol, Div Basic Sci, Frederick, MD 21702 USA. RP Corsi, MM (reprint author), Univ Milan, Fac Med, Inst Gen Pathol, I-20133 Milan, Italy. NR 21 TC 7 Z9 7 U1 0 U2 0 PU AMER ASSOC CLINICAL CHEMISTRY PI WASHINGTON PA 2101 L STREET NW, SUITE 202, WASHINGTON, DC 20037-1526 USA SN 0009-9147 J9 CLIN CHEM JI Clin. Chem. PD JUN PY 2001 VL 47 IS 6 BP 1135 EP 1137 PN 1 PG 3 WC Medical Laboratory Technology SC Medical Laboratory Technology GA 437MW UT WOS:000168996300035 PM 11375314 ER PT J AU Cecco, SA Rehak, NN AF Cecco, SA Rehak, NN TI Short-term and long-term storage of plasma in gel separator tubes: Effect on common chemistry tests determined with Hitachi 917. SO CLINICAL CHEMISTRY LA English DT Meeting Abstract C1 NIH, Bethesda, MD 20892 USA. NR 0 TC 0 Z9 0 U1 0 U2 0 PU AMER ASSOC CLINICAL CHEMISTRY PI WASHINGTON PA 2101 L STREET NW, SUITE 202, WASHINGTON, DC 20037-1526 USA SN 0009-9147 J9 CLIN CHEM JI Clin. Chem. PD JUN PY 2001 VL 47 IS 6 SU S MA 293 BP A89 EP A89 PN 2 PG 1 WC Medical Laboratory Technology SC Medical Laboratory Technology GA 437MX UT WOS:000168996400409 ER EF