TY - BOOK T1 - The year in rheumatic disorders 2002: special issue: rheumatoid arthritis A1 - Cope AP Y1 - 2002/// SN - 0-9537-3399-8 N2 - - ER - TY - BOOK T1 - Pocket Reference to TNF alpha antagonism and rheumatoid arthritis A1 - Maini RN A1 - Feldmann M Y1 - 2000/// PB - Science Press Ltd CY - London N2 - - ER - TY - CHAP T1 - Mapping lymphocyte plasma membrane proteins: A proteomic approach A1 - Peirce M J A1 - Saklatvala J A1 - Cope A P A1 - Wait R ED - AP Cope T2 - Arthritis Research: Y1 - 2007/// M2 - 136 PB - Humana Press N2 - - ER - TY - CHAP T1 - Internal standards in differentiating embryonic stem cells in vitro. A1 - Murphy, CL T2 - Methods in Molecular Biology Y1 - 2006/07/19/ M2 - 329 PB - Humana Press CY - United States SP - 101 EP - 112 N2 - - UR - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?CMD=Display&DB=pubmed ER - TY - CHAP T1 - Toll-like receptors and rheumatoid arthritis: is there a connection? A1 - Sacre SM A1 - Drexler S A1 - Foxwell BM ED - O'Neill L; Brint E T2 - Toll-like Receptors in Inflammation Y1 - 2005/12// PB - Birkhauser SN - 3-7643-7285-0 N2 - - ER - TY - CHAP T1 - Quantitative Profiling of Protein-DNA Binding on Microarrays A1 - Ragoussis J A1 - Field S A1 - Udalova IA ED - Bina M T2 - Methods in Molecular Biology, vol. 338: Gene Mapping, Discovery, and Expression: Methods and Protocols Y1 - 2005/// PB - Humana Press Inc CY - USA N2 - - ER - TY - CHAP T1 - Immunopathogenesis of rheumatoid arthritis A1 - Maini RN A1 - Feldmann M ED - Isenberg DA; Maddison PJ; Woo P; Glass D; Breedveld FC T2 - Oxford Textbook of Rheumatology Y1 - 2004/// M2 - 4th Ed PB - Oxford University Press CY - Oxford SN - 0 19 850948 0 SP - 677 EP - 697 N2 - - ER - TY - CHAP T1 - The transfer of a laboratory based hypothesis to a clinically useful therapy: the development of anti-TNF therapy of rheumatoid arthritis. A1 - Feldmann M A1 - Brennan FM A1 - WIlliams RO A1 - Woody JN A1 - Maini RN ED - L. Klareskog and S. Lindblad T2 - Scientific Basis of Rheumatology: Best Practice & Research, Clinical Rheumatology Y1 - 2004/// M2 - 18 (1) PB - Bailliere Tindall SP - 59 EP - 80 N2 - - ER - TY - CHAP T1 - The role of tumour necrosis factor in rheumatoid arthritis A1 - Maini RN ED - van den Berg WB; Miossec P T2 - Cytokines and Injury in Progress in Inflammation Research Y1 - 2004/// PB - Birkhauser Verlag CY - Basel SN - 3-7643-7001-7 SP - 1 EP - 28 N2 - - ER - TY - CHAP T1 - The Role of NF-kB in Inflammatory Diseases A1 - Andreakos E A1 - Udalova I A1 - Sacre S A1 - Foxwell BM ED - Beyaert, R T2 - Nuclear Factor Kb: Regulation and Role in Disease Y1 - 2003/07// PB - Kluwer Acad. Publrs CY - Netherlands SN - 1-4020-1434-1 N2 - - UR - http://www.springeronline.com/sgw/cda/frontpage/0,11855,5-128-22-33495140-detailsPage%253Dppmmedia%257CaboutThisBook%257CaboutThisBook,00.html ER - TY - CHAP T1 - TNFalpha A1 - Feldmann M A1 - Maini RN. ED - J. Smolen and P. Lipsky T2 - Targeted Therapies in Rheumatology Y1 - 2003/// PB - Martin Dunitz Ltd. CY - London SP - 201 EP - 212 N2 - - ER - TY - CHAP T1 - Infliximab therapy. A1 - Maini RN A1 - Feldmann M ED - L.W. Moreland and P. Emery T2 - TNFalpha inhibition in the treatment of rheumatoid arthritis. Y1 - 2003/// PB - Martin Dunitz CY - London SP - 23 EP - 45 N2 - - ER - TY - CHAP T1 - The role of NF-kappaB in inflammatory diseases. A1 - Andreakos E A1 - Udalova IA A1 - Sacre S ED - Beyaert R T2 - Nuclear Factor kappaB. Regulation and Role in Disease. Y1 - 2003/// PB - Kluwer Academic Publishers CY - Netherlands SP - 297 EP - 325 N2 - - ER - TY - CHAP T1 - Definition of TNFalpha as a therapeutic target for rheumatoid arthritis A1 - Feldmann M A1 - Brennan FM A1 - Williams RO A1 - Maini RN ED - Moreland LW; Emery P T2 - TNFalpha-inhibition in the treatment of rheumatoid arthritis Y1 - 2003/// PB - Martin Dunitz CY - London SN - 1-84184-156-0 SP - 1 EP - 22 N2 - - ER - TY - CHAP T1 - Assessment of disease activity in rheumatoid arthritis A1 - Abrahams S A1 - Cope A A1 - Taylor PC ED - Scott D and Cope A T2 - The year in rheumatic disorders 2003 Y1 - 2003/// PB - Clinical Publishing Services CY - Oxford SN - 1 904392 09 1 SP - 103 EP - 126 N2 - - ER - TY - CHAP T1 - Definition of TNFalpha as a therapeutic target for rheumatoid arthritis A1 - Feldmann M A1 - Brennan F A1 - Williams RO A1 - Maini RN ED - Moreland, Emery T2 - TNFalpha inhibition in the treatment of rheumatoid arthritis Y1 - 2003/// PB - Martin Dunitz CY - London SP - 1 EP - 22 N2 - - ER - TY - CHAP T1 - TNF alpha A1 - Feldmann M A1 - Maini RN ED - Smolen J; Lipsky P T2 - Targeted Therapies in Rheumatology Y1 - 2003/// PB - Martin Dunitz Ltd CY - London SN - 1-84184-157-9 SP - 201 EP - 212 N2 - - ER - TY - CHAP T1 - Autoimmunity and cytokines: pathogenesis and therapy. A1 - Andreakos E A1 - Feldmann M ED - A.W. Thomson and M.T. Lotze T2 - Cytokine Handbook, 4th Edition Y1 - 2003/// PB - Academic Press SP - 1189 EP - 1211 N2 - - ER - TY - CHAP T1 - Rheumatoid Arthritis A1 - Maini RN ED - Warrell DA; Cox TM; Firth JD T2 - Oxford Textbook of Medicine Y1 - 2003/// M2 - 4th edition PB - Oxford University Press CY - Oxford SN - 0-19-262922-0 N2 - - ER - TY - CHAP T1 - Infliximab Therapy A1 - Feldmann M A1 - Maini RN ED - Moreland LW; Emery P T2 - TNFalpha-inhibition in the treatment of rheumatoid arthritis Y1 - 2003/// PB - Martin Dunitz CY - London SN - 1-84184-156-0 SP - 23 EP - 45 N2 - - ER - TY - CHAP T1 - Definition of TNFalpha as a therapeutic target for rheumatoid arthritis. A1 - Feldmann M A1 - Brennan FM A1 - Williams RO A1 - Maini RN ED - L.W. Moreland and P. Emery T2 - TNFalpha inhibition in the treatment of rheumatoid arthritis. Y1 - 2003/// PB - Martin Dunitz CY - London SP - 1 EP - 22 N2 - - ER - TY - CHAP T1 - Angiogenesis A1 - Paleolog EM T2 - The year in rheumatic disorders 2002: special issue: rheumatoid arthritis Y1 - 2002/// SN - 0-9537-3399-8 SP - 63 EP - 80 N2 - - ER - TY - CHAP T1 - Gene therapy approaches for rheumatoid arthritis A1 - Bondeson J A1 - Feldmann M A1 - Maini RN T2 - Gene Therapy: the use of DNA as a drug Y1 - 2002/// SN - 0-8536-9455-9 SP - 215 EP - 265 N2 - - ER - TY - CHAP T1 - Imaging A1 - Taylor P T2 - The year in rheumatic disorders 2002: special issue: rheumatoid arthritis Y1 - 2002/// SN - 0-9537-3399-8 SP - 159 EP - 177 N2 - - ER - TY - CHAP T1 - Angiogenesis A1 - Paleolog EM T2 - The year in rheumatic disorders 2002: special issue: rheumatoid arthritis Y1 - 2002/// SN - 0-9537-3399-8 SP - 63 EP - 80 N2 - - ER - TY - CHAP T1 - Imaging A1 - Taylor P T2 - The year in rheumatic disorders 2002: special issue: rheumatoid arthritis Y1 - 2002/// SN - 0-9537-3399-8 SP - 159 EP - 177 N2 - - ER - TY - CHAP T1 - Lymphocyte biology A1 - Cope A T2 - The Year in Rheumatic Disorders 2002: special issue: rheumatoid arthritis Y1 - 2002/// SN - 0-9537-3399-8 SP - 81 EP - 106 N2 - - ER - TY - CHAP T1 - Gene therapy approaches for rheumatoid arthritis A1 - Bondeson J A1 - Feldmann M A1 - Maini RN T2 - Gene Therapy: the use of DNA as a drug Y1 - 2002/// SN - 0-8536-9455-9 SP - 215 EP - 265 N2 - - ER - TY - CHAP T1 - Proinflammatory cytokines A1 - Feldmann M A1 - Saklatvala J Y1 - 2001/// SN - 0-1225-2670-8 SP - 291 EP - 305 N2 - - ER - TY - CHAP T1 - Exploring the pathogenesis of rheumatoid arthritis in transgenic and mutant mice A1 - Cope AP Y1 - 2001/// SN - 3-8055-7120-8 SP - 64 EP - 93 N2 - - ER - TY - CHAP T1 - Metalloproteinases A1 - Nagase H Y1 - 2001/// SN - 0-4711-1184-8 SP - 21 EP - 21 N2 - - ER - TY - CHAP T1 - Cytokines and disease A1 - Feldmann M A1 - Brennan FM Y1 - 2001/// SN - 0-1225-2670-8 SP - 35 EP - 51 N2 - - ER - TY - CHAP T1 - TNF receptors A1 - Aggarwal BB A1 - Samanta A A1 - Feldmann M Y1 - 2001/// SN - 0-1225-2670-8 SP - 1619 EP - 1632 N2 - - ER - TY - CHAP T1 - TNF? A1 - Aggarwal BB A1 - Samanta A A1 - Feldmann M Y1 - 2001/// SN - 0-1225-2670-8 SP - 413 EP - 434 N2 - - ER - TY - CHAP T1 - Finding, purification and characterization of natural protease inhibitors A1 - Nagase H A1 - Salvesen GS Y1 - 2001/// M2 - 2nd SN - 0-1996-3663-X SP - 131 EP - 147 N2 - - ER - TY - CHAP T1 - Introduction to the role of cytokines in innate host defense and adaptive immunity A1 - Oppenheim JJ A1 - Feldmann M Y1 - 2001/// SN - 0-1225-2670-8 SP - 3 EP - 20 N2 - - ER - TY - CHAP T1 - The role of TNF-Alpha and IL-1 in rheumatoid arthritis A1 - Feldmann M A1 - Brennan FM A1 - Foxwell BMJ A1 - Maini RN Y1 - 2001/// SN - 3-8055-7120-8 SP - 188 EP - 199 N2 - - ER - TY - CHAP T1 - The role of TNF? and IL-1 in rheumatoid arthritis A1 - Feldmann M A1 - Brennan FM A1 - Foxwell BMJ A1 - Maini RN Y1 - 2001/// SN - 3-8055-7120-8 SP - 188 EP - 199 N2 - - ER - TY - CHAP T1 - Cytokines and disease A1 - Feldmann M A1 - Brennan FM Y1 - 2001/// SN - 0-1225-2670-8 SP - 35 EP - 51 N2 - - ER - TY - CHAP T1 - The role of TNF? and IL-1 in rheumatoid arthritis A1 - Feldmann M A1 - Brennan FM A1 - Foxwell BMJ A1 - Maini RN Y1 - 2001/// SN - 3-8055-7120-8 SP - 188 EP - 199 N2 - - ER - TY - CHAP T1 - Donor lymphocyte infusions A1 - Dazzi F A1 - Stauss HJ Y1 - 2001/// SN - 1-8531-7890-X SP - 385 EP - 399 N2 - - ER - TY - CHAP T1 - Proinflammatory cytokines A1 - Feldmann M A1 - Saklatvala J Y1 - 2001/// SN - 0-1225-2670-8 SP - 291 EP - 305 N2 - - ER - TY - CHAP T1 - Substrate specificity of MMPs A1 - Nagase H Y1 - 2001/// SN - 0-8960-3668-5 SP - 39 EP - 66 N2 - - ER - TY - CHAP T1 - Inhibition of proteolytic enzymes A1 - Salvesen GS A1 - Nagase H Y1 - 2001/// M2 - 2nd SN - 0-1996-3662-1 SP - 105 EP - 130 N2 - - ER - TY - CHAP T1 - The role of TNFalpha and IL-1 in rheumatoid arthritis A1 - Feldmann M A1 - Brennan FM A1 - Foxwell BMJ A1 - Maini RN Y1 - 2001/// SN - 3-8055-7120-8 SP - 188 EP - 199 N2 - - ER - TY - CHAP T1 - Anti-TNFalpha therapy in rheumatoid arthritis and other diseases. A1 - Feldmann M A1 - Maini RN ED - Austen KF; Frank MM; Atkinson JP; Cantor H T2 - Samter's Immunologic Diseases (Sixth Edition) Y1 - 2001/// PB - Lippincott Williams & Wilkins SP - 1180 EP - 1198 N2 - - ER - TY - CHAP T1 - The debut of anti-TNF therapy of rheumatoid arthritis in the clinic A1 - Maini RN ED - Higgs GA; Henderson B T2 - Progress in Inflammation Research Y1 - 2000/// PB - Birkhauser Verlag CY - Basel SP - 145 EP - 156 N2 - - ER - TY - CHAP T1 - The role of protein phosphatases in cell signalling by the high affinity receptor for immunoglobulin E A1 - Peirce M J ED - Ehud Razin and Juan Rivera T2 - Signal transduction in mast cells and basophils Y1 - 1999/// PB - Springer-Verlag CY - New York SN - 0-387-98625-1 SP - 134 EP - 151 N2 - - ER - TY - CONF T1 - Resuscitation of thermal injuries in the United Kingdom and Ireland A1 - Baker, RHJ A1 - Akhavani, MA A1 - Jallali, N U1 - 38th Annual Meeting of the British-Burns-Association Y1 - 2007/// Y2 - // VL - 60 SP - 682 EP - 685 N2 - - ER - TY - CONF T1 - Bench to Bedside: Anti-TNF a paradigm for biological targeted therapy A1 - Maini RN U1 - Therapeutic antibodies - 9th Annual Symposium - Ranbaxy Science Foundation AD - New Delhi Y1 - 2003/// Y2 - 2003/// PB - Ranbaxy Science Foundation SP - 13 EP - 24 N2 - - ER - TY - JFULL T1 - Anti-inflammatory functions of glucocorticoid-induced genes. A1 - Clark, AR J1 - Mol Cell Endocrinol Y1 - 2007/09/15/ VL - 275 SN - 0303-7207 SP - 79 EP - 97 N2 - There is a broad consensus that glucocorticoids (GCs) exert anti-inflammatory effects largely by inhibiting the function of nuclear factor kappaB (NFkappaB) and consequently the transcription of pro-inflammatory genes. In contrast, side effects are thought to be largely dependent on GC-induced gene expression. Biochemical and genetic evidence suggests that the positive and negative effects of GCs on transcription can be uncoupled from one another. Hence, novel GC-related drugs that mediate inhibition of NFkappaB but do not activate gene expression are predicted to retain therapeutic effects but cause fewer or less severe side effects. Here, we critically re-examine the evidence in favor of the consensus, binary model of GC action and discuss conflicting evidence, which suggests that anti-inflammatory actions of GCs depend on the induction of anti-inflammatory mediators. We propose an alternative model, in which GCs exert anti-inflammatory effects at both transcriptional and post-transcriptional levels, both by activating and inhibiting expression of target genes. The implications of such a model in the search for safer anti-inflammatory drugs are discussed. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17561338&query_hl=1 ER - TY - JFULL T1 - The anti-allergic drug, N-(3',4'-dimethoxycinnamonyl) anthranilic acid, exhibits potent anti-inflammatory and analgesic properties in arthritis. A1 - Inglis, JJ A1 - Criado, G A1 - Andrews, M A1 - Feldmann, M A1 - Williams, RO A1 - Selley, ML J1 - Rheumatology (Oxford) Y1 - 2007/09// VL - 46 SN - 1462-0324 SP - 1428 EP - 1432 N2 - Objectives. The degradation of tryptophan by indoleamine 2,3-dioxygenase yields a number of immunomodulatory metabolites, including 3-hydroxyanthranilic acid, 3-hydroxykynurenic acid and quinolinic acid. N-(3',4'-dimethoxycinnamonyl) anthranilic acid (3,4-DAA) is a synthetic anthranilic acid derivative that has been used therapeutically in Japan for many years as an anti-allergic drug and has recently been shown to be effective in a murine model of multiple sclerosis. Methods. In the present study, we tested the efficacy of 3,4-DAA in collagen-induced arthritis, a mouse model of rheumatoid arthritis, and analysed its mechanism of action. Results. Administration of 3,4-DAA after arthritis onset reduced clinical and histological severity of arthritis and reduced pain. It completely abrogated thermal and mechanical hyperalgesia. 3,4-DAA also suppressed Th1 cell activity in lymph node cell cultures and raised serum levels of IL-10. In vitro, 3,4-DAA suppressed IFNgamma production and proliferation of both T and B lymphocytes in a manner comparable with the endogenous tryptophan metabolite, 3-hydroxyanthranilic acid, suggesting similar mechanisms of action. Conclusion. It is concluded that 3,4-DAA has both anti-inflammatory and analgesic properties, and may therefore be useful in filling an unmet need, in the treatment of rheumatoid and other forms of arthritis, especially in the light of its analgesic properties. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17644821&query_hl=1 ER - TY - JFULL T1 - Mass Spectrometric Analysis of the Endogenous Type I Interleukin-1 (IL-1) Receptor Signaling Complex Formed after IL-1 Binding Identifies IL-1RAcP, MyD88, and IRAK-4 as the Stable Components. A1 - Brikos, C A1 - Wait, R A1 - Begum, S A1 - O'neill, LA A1 - Saklatvala, J J1 - Mol Cell Proteomics Y1 - 2007/09// VL - 6 SN - 1535-9476 SP - 1551 EP - 1559 N2 - We investigated the composition of the endogenous ligand-bound type I interleukin-1 (IL-1) receptor (IL-1RI) signaling complex using immunoprecipitation and tandem mass spectrometry. Three proteins with approximate molecular masses of 60 (p60), 36 (p36), and 90 kDa (p90) became phosphorylated after treatment with IL-1. Phosphorylation in vitro of p60 has been reported previously, but its identity was unknown. We showed using tandem mass spectrometry that p60 is identical to interleukin-1 receptor-associated kinase (IRAK)-4. MS also enabled detection of IL-1, IL-1RI, IL-1 receptor accessory protein (IL-1RAcP), and myeloid differentiation primary response protein 88 (MyD88) in the complex. The p60 protein (IRAK-4) was the earliest component of the complex to be phosphorylated. Phosphorylated IRAK-4 from the receptor complex migrated more slowly in SDS-PAGE than its unphosphorylated form as did recombinant IRAK-4 autophosphorylated in vitro. Phosphorylation was restricted to serine and threonine residues. IRAK-4, p36, IL-1RAcP, and MyD88 bound to the liganded receptor within 15 s of activation by IL-1 and remained associated upon prolonged activation, suggesting that the signaling complex is very stable. The p90 phosphoprotein was only transiently associated with the receptor. This behavior and its size were consistent with it being IRAK-1. Our work revealed that liganding of IL-1RI causes its strong and stable association with IL-1RAcP, MyD88, and the previously unidentified protein p60 (IRAK-4). The only component of the IL-1RI signaling complex that dissociated is IRAK-1. Our study is therefore the first detailed description of the endogenous IL-1RI complex. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17507369&query_hl=1 ER - TY - JFULL T1 - Mesenchymal stem cells of cord blood origin are effective at preventing but not treating graft-versus-host disease. A1 - Tisato, V A1 - Naresh, K A1 - Girdlestone, J A1 - Navarrete, C A1 - Dazzi, F J1 - Leukemia Y1 - 2007/09// VL - 21 SN - 0887-6924 SP - 1992 EP - 1999 N2 - The immunosuppressive properties of mesenchymal stem cells (MSC) make them particularly attractive to manipulate graft-versus-host disease (GVHD). So far, the experience of using MSC to treat GVHD is limited to a few cases, controversial results come from preclinical models and several issues remain to be clarified. The present studies were designed to address these questions in a xenogenic model testing the ability of umbilical cord blood-derived MSC (UCB-MSC) to prevent and/or treat GVHD. Sublethally irradiatiated non-obese diabetic/severe combined immunodeficiency NOD/SCID mice transplanted with human peripheral blood mononuclear cells (huPBMC) showed extensive human T-cell proliferation in the peripheral blood, lymphoid and non-lymphoid tissues, which evolved in extensive GVHD (wasting, ruffled hair and hunched back). The mice treated with a single dose of UCB-MSC did not behave differently form the controls. However, when UCB-MSC were given at weekly intervals, there was a marked decrease in human T-cell proliferation and none of the mice developed GVHD. No therapeutic effect was obtained if UCB-MSC were administered at onset of GVHD. This work supports the clinical use of MSC in stem cell transplantation as a prophylaxis rather than treatment of GVHD.Leukemia (2007) 21, 1992-1999; doi:10.1038/sj.leu.2404847; published online 12 July 2007. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17625609&query_hl=1 ER - TY - JFULL T1 - Protein Sulfenation as a Redox Sensor: Proteomics Studies Using a Novel Biotinylated Dimedone Analogue. A1 - Charles, RL A1 - Schröder, E A1 - May, G A1 - Free, P A1 - Gaffney, PR A1 - Wait, R A1 - Begum, S A1 - Heads, RJ A1 - Eaton, P J1 - Mol Cell Proteomics Y1 - 2007/09// VL - 6 SN - 1535-9476 SP - 1473 EP - 1484 N2 - Protein sulfenic acids are reactive intermediates in the catalytic cycles of many enzymes as well as the in formation of other redox states. Sulfenic acid formation is a reversible post-translational modification with potential for protein regulation. Dimedone (5,5-dimethyl-1,3-cyclohexanedione) is commonly used in vitro to study sulfenation of purified proteins, selectively "tagging" them, allowing monitoring by mass spectrometry. However dimedone is of little use in complex protein mixtures because selective monitoring of labeling is not possible. To address this issue, we synthesized a novel biotinylated derivative of dimedone, keeping the dione cassette required for sulfenate reactivity but adding the functionality of a biotin tag. Biotin-amido(5-methyl-5-carboxamidocyclohexane 1,3-dione) tetragol (biotin dimedone) was prepared in six steps, combining 3,5-dimethoxybenzoic acid (Birch reduction, ultimately leading to the dimedone unit with a carboxylate functionality), 1-amino-11-azido-3,6,9-trioxaundecane (a differentially substituted tetragol spacer), and biotin. We loaded biotin dimedone (0.1 mm, 30 min) into rat ventricular myocytes, treated them with H(2)O(2) (0.1-10,000 mum, 5 min), and monitored derivatization on Western blots using streptavidin-horseradish peroxidase. There was a dose-dependent increase in labeling of multiple proteins that was maximal at 0.1 or 1 mm H(2)O(2) and declined sharply below basal with 10 mm treatment. Cell-wide labeling was observed in fixed cells probed with avidin-FITC using a confocal fluorescence microscope. Similar H(2)O(2)-induced labeling was observed in isolated rat hearts. Hearts loaded and subjected to hypoxia showed a striking loss of labeling, which returned when oxygen was resupplied, highlighting the protein sulfenates as oxygen sensors. Cardiac proteins that were sulfenated during oxidative stress were purified with avidin-agarose and identified by separation of tryptic digests by liquid chromatography with on-line analysis by mass spectrometry. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17569890&query_hl=1 ER - TY - JFULL T1 - Reply to 'Splenectomy status of the patient may have impact on response to donor lymphocyte infusions' by Duygu Uckan-Cetinkaya et al. A1 - Dazzi, F A1 - Fozza, C J1 - Leukemia Y1 - 2007/09// VL - 21 SN - 0887-6924 SP - 2050 EP - 2051 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17597812&query_hl=1 ER - TY - JFULL T1 - Regulation of fibroblast migration by tenascin-C. A1 - Trebaul, A A1 - Chan, EK A1 - Midwood, KS J1 - Biochem Soc Trans Y1 - 2007/08// VL - 35 SN - 0300-5127 SP - 695 EP - 697 N2 - Synthesis of new tissue by fibroblasts is required for tissue rebuilding in response to injury. Fibroblast migration from surrounding healthy tissue into the fibrin-fibronectin provisional matrix deposited upon injury is a key rate-limiting step of this stage of tissue repair. These events must be tightly regulated. Excessive deposition of scar tissue is the major hallmark of fibrotic disease. Tenascin-C is an extracellular matrix glycoprotein that is transiently expressed upon tissue injury, where it is specifically localized to the wound edge, and persistently up-regulated in fibrotic disease. We have shown that full-length tenascin-C promotes fibroblast migration within fibrin-fibronectin matrices and we have mapped the domains within the molecule critical for enhancing migration. We also demonstrated that specific fragments of tenascin-C inhibit fibroblast migration. These results suggest that transient expression of tenascin-C at the wound boundary is key to tissue repair: its induction recruits fibroblasts into the wound and fragments resulting from its breakdown prevent excessive fibroblast infiltration. Our results demonstrate how fibroblast migration in three-dimensional provisional matrices may be differentially regulated by proteolysis of matrix molecules and could explain how persistent expression of tenascin-C contributes to the progression of fibrotic disease. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17635125&query_hl=1 ER - TY - JFULL T1 - Cytokine inhibitors in rheumatoid arthritis and other autoimmune diseases. A1 - Williams, RO A1 - Paleolog, E A1 - Feldmann, M J1 - Curr Opin Pharmacol Y1 - 2007/08// VL - 7 SN - 1471-4892 SP - 412 EP - 417 N2 - The clinical success of TNFalpha blocking biologics in a growing number of immune-mediated pathologies, including rheumatoid arthritis, Crohn's disease, ankylosing spondylitis and psoriasis, confirms the importance of TNFalpha in driving chronic inflammation and represents an important step forward in the treatment of these conditions. TNFalpha blockade, however, is a treatment, rather than a cure, and is not effective in all patients or in all autoimmune diseases and further research is needed to get closer to a cure. Recently, the identification of a novel, IL-17 producing, T helper cell subset, that plays a dominant pathogenic role in animal models of autoimmunity, is a major advance on existing knowledge, although the role of these cells in human disease remains to be established. Cytokines driving angiogenesis are also important in disease chronicity and thus might be valid therapeutic targets. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17627887&query_hl=1 ER - TY - JFULL T1 - Key differences in TLR3/POLY I:C signaling and cytokine induction by human primary cells: a phenomenon absent from murine cell systems. A1 - Lundberg, AM A1 - Drexler, SK A1 - Monaco, C A1 - Williams, LM A1 - Sacre, SM A1 - Feldmann, M A1 - Foxwell, BM J1 - Blood Y1 - 2007/07/27/ SN - 0006-4971 N2 - TLR3 recognizes double-stranded RNA, a product associated with viral infections. Many details of TLR3-induced mechanisms have emerged from gene-targeted mice or inhibition studies in transformed cell lines. However, the pathways activated in human immune cells or cells from disease tissue are less well understood. We have investigated TLR3-induced mechanisms of human primary cells of the innate immune system, including dendritic cells (DC), macrophages (MØ), endothelial cells (EC) and synovial fibroblasts isolated from rheumatoid arthritis joint tissue (RA-SF). Here we report that while these cells all express TLR3 they differ substantially in their response to TLR3 stimulation. The key antiviral response chemokine IP-10 was produced by all cell types, while DC and MØ failed to produce the proinflammatory cytokines TNFalpha and IL-6. Unexpectedly, TNFalpha was found secreted by TLR3 stimulated RA-SF. Furthermore, TLR3 stimulation did not activate NFkappaB, MAPKs or IRF-3 in DC and MØ, but were able to do so in EC and RA-SF. These findings were specific for human cells thereby revealing a complexity not previously expected. This is the first report of such cell type and species-specific response for any TLR stimulation and helps understand important difficulties in correlating murine models of inflammatory diseases and human inflammation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17660379&query_hl=1 ER - TY - JFULL T1 - FGF-2 is bound to perlecan in the pericellular matrix of articular cartilage, where it acts as a chondrocyte mechanotransducer. A1 - Vincent, TL A1 - McLean, CJ A1 - Full, LE A1 - Peston, D A1 - Saklatvala, J J1 - Osteoarthritis Cartilage Y1 - 2007/07// VL - 15 SN - 1063-4584 SP - 752 EP - 763 N2 - OBJECTIVE: We have shown previously that cutting or loading articular cartilage resulted in a fibroblast growth factor-2 (FGF-2) dependent activation of the extracellularly regulated kinase (ERK), and induction of a number of chondrocyte regulatory proteins including tissue inhibitor of metalloproteinase-1 and matrix metalloproteinases 1 and 3. An extracellular matrix-bound pool of FGF-2 was apparent, which could be liberated from the tissue by heparitinase (Vincent et al., Proc Natl Acad Sci U S A 2002;99(12):8259-64, Vincent et al., Arthritis Rheum 2004 Feb;50(2):526-33). Our objectives were to determine where FGF-2 was stored in articular cartilage, to which proteoglycan it was bound, and to elucidate its role in chondrocyte mechanotransduction. METHODS: Immunohistochemistry and confocal microscopy were used to localise FGF-2 in the tissue. In vitro binding studies were performed using IASYS surface plasmon resonance. To study the role of pericellular FGF-2 in mechanotransduction cartilage explants or articular chondrocytes encapsulated in alginate were loaded using an in house loading rig. The loading response was assessed by the activation of ERK, in the presence or absence of a specific FGFR inhibitor. RESULTS: Here we have identified perlecan as the heparan sulphate proteoglycan that sequesters FGF-2 in articular cartilage. Perlecan and FGF-2 co-localised within the type VI collagen-rich pericellular matrix of porcine and human articular cartilage. Chondrocytes encapsulated in alginate were able to accumulate pericellular perlecan and FGF-2 in culture, and deliver an FGF-dependent activation of ERK when loaded. CONCLUSION: Loading-induced ERK activation was dependent upon the presence and concentration of pericellular FGF-2, suggesting a functional role for this matrix-bound growth factor in chondrocyte mechanotransduction. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17368052&query_hl=1 ER - TY - JFULL T1 - Insertional polymorphisms: a new lease of life for endogenous retroviruses in human disease. A1 - Moyes, D A1 - Griffiths, DJ A1 - Venables, PJ J1 - Trends Genet Y1 - 2007/07// VL - 23 SN - 0168-9525 SP - 326 EP - 333 N2 - Human endogenous retroviruses (HERVs) result from ancestral infection by infectious viruses over millions of years of primate evolution. Some are transcriptionally active, express proteins and therefore have the potential to cause disease. Here we review the controversial attempts to link them with cancer and autoimmunity. The main difficulty is that most HERVs investigated to date are present at the same locus in 100% of the population. However, a new class of insertionally polymorphic HERV-K family members, present in a minority of individuals, has recently been described. We propose that insertionally polymorphic HERVs could be novel genetic risk factors and hence provide a new lease of life for research into HERVs and disease. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17524519&query_hl=1 ER - TY - JFULL T1 - Proteolytic activities of human ADAMTS-5: comparative studies with ADAMTS-4. A1 - Gendron, C A1 - Kashiwagi, M A1 - Lim, NH A1 - Enghild, JJ A1 - Thøgersen, IB A1 - Hughes, C A1 - Caterson, B A1 - Nagase, H J1 - J Biol Chem Y1 - 2007/06/22/ VL - 282 SN - 0021-9258 SP - 18294 EP - 18306 N2 - Aggrecanases have been characterized as proteinases that cleave the Glu373-Ala374 bond of the aggrecan core protein, and they are multidomain metalloproteinases belonging to the ADAMTS (adamalysin with thrombospondin type 1 motifs) family. The first aggrecanases discovered were ADAMTS-4 (aggrecanase 1) and ADAMTS-5 (aggrecanase 2). They contain a zinc catalytic domain followed by non-catalytic ancillary domains, including a disintegrin domain, a thrombospondin domain, a cysteine-rich domain, and a spacer domain. In the case of ADAMTS-5, a second thrombospondin domain follows the spacer domain. We previously reported that the non-catalytic domains of ADAMTS-4 influence both its extracellular matrix interaction and proteolytic abilities. Here we report the effects of these domains of ADAMTS-5 on the extracellular matrix interaction and proteolytic activities and compare them with those of ADAMTS-4. Although the spacer domain was critical for ADAMTS-4 localization in the matrix, the cysteine-rich domain influenced ADAMTS-5 localization. Similar to previous reports of other ADAMTS family members, very little proteolytic activity was detected with the ADAMTS-5 catalytic domain alone. The sequential inclusion of each carboxyl-terminal domain enhanced its activity against aggrecan, carboxymethylated transferrin, fibromodulin, decorin, biglycan, and fibronectin. Both ADAMTS-4 and -5 had a broad optimal activity at pH 7.0-9.5. Aggrecanolytic activities were sensitive to the NaCl concentration, but activities on non-aggrecan substrates, e.g. carboxymethylated transferrin, were not affected. Although ADAMTS-4 and ADAMTS-5 had similar general proteolytic activities, the aggrecanase activity of ADAMTS-5 was at least 1,000-fold greater than that of ADAMTS-4 under physiological conditions. Our studies suggest that ADAMTS-5 is a major aggrecanase in cartilage metabolism and pathology. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17430884&query_hl=1 ER - TY - JFULL T1 - Prevalence of human papillomavirus in skin tumors from repair deficient xeroderma pigmentosum patients. A1 - Luron, L A1 - Avril, MF A1 - Sarasin, A A1 - Daya-Grosjean, L J1 - Cancer Lett Y1 - 2007/06/08/ VL - 250 SN - 0304-3835 SP - 213 EP - 219 N2 - The predisposition to skin cancers in childhood is the hallmark of xeroderma pigmentosum (XP), a rare autosomal recessive disorder, deficient in DNA repair and hypersensitive to ultraviolet irradiation. Human papillomavirus (HPVs), are common infections of the skin which are often found associated to benign lesions and non-melanoma skin cancers (NMSC), mainly squamous cell carcinomas (SCC) and basal cell carcinomas (BCC). Our study is the first to analyse 40 SCCs, 27 BCCs and nine normal skin biopsies from XP patients for HPV DNA which are found more frequently in SCCs (20/40) than in BCCs (4/27) or normal skin (2/9). The HPV spectrum includes 22 different epidermodysplasia verruciformis (EV) HPV types, which predominate in SCCs (48%) compared to BCCs (15%) and normal skin (22%). Our data, showing an association between EV HPV and SCCs from young XP patients is comparable to that found for NMSC from adult immunosuppressed organ transplant patients and raises the question of the importance of HPV infection in skin carcinogenesis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17126994&query_hl=1 ER - TY - JFULL T1 - Low intensity transplantat regimens facilitate recruitment of donor apecific regulatory T cell which promote heamatopoietic engraftment A1 - Weng, L A1 - Dyson, J A1 - Dazzi, F J1 - HAEMATOL-HEMATOL J Y1 - 2007/06// VL - 92 SN - 0390-6078 SP - 379 EP - 379 ER - TY - JFULL T1 - Do Tc-99m-diphosphonate bone scans have any place in the investigation of polyarthralgia? A1 - Fisher, BA A1 - Frank, JW A1 - Taylor, PC J1 - Rheumatology (Oxford) Y1 - 2007/06// VL - 46 SN - 1462-0324 SP - 1036 EP - 1037 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17449485&query_hl=1 ER - TY - JFULL T1 - NF-kappa B inhibition by blockade of IKK-2 and MYD88 in vein graft smooth muscle cells reduces the expression of key mediators of intimal hyperplasia A1 - Finch, J A1 - Navin, T A1 - Gregan, S A1 - Foxwell, B A1 - Haskard, D A1 - Monaco, C A1 - Hornick, P J1 - HEART Y1 - 2007/06// VL - 93 SN - 1355-6037 SP - A24 EP - A25 ER - TY - JFULL T1 - Cord blood derived mesenchymal stem cells are effective at preventing graft-versus-host disease A1 - Tisato, V A1 - Naresh, K A1 - Girdlestone, J A1 - Navarrete, C A1 - Dazzi, F J1 - HAEMATOL-HEMATOL J Y1 - 2007/06// VL - 92 SN - 0390-6078 SP - 317 EP - 317 ER - TY - JFULL T1 - Mesenchymal stem cell mediated immunosuppression is not confined to progenitor status A1 - Jones, F A1 - Horwood, N A1 - Cope, A A1 - Dazzi, F J1 - HAEMATOL-HEMATOL J Y1 - 2007/06// VL - 92 SN - 0390-6078 SP - 316 EP - 316 ER - TY - JFULL T1 - Disease relapse after haematopoietic stem cell transplantation: risk factors and treatment. A1 - Dazzi, F A1 - Fozza, C J1 - Best Pract Res Clin Haematol Y1 - 2007/06// VL - 20 SN - 1521-6926 SP - 311 EP - 327 N2 - Disease relapse is the commonest cause of treatment failure after allogeneic haematopoietic stem-cell transplantation. Adoptive immunotherapy based on donor lymphocyte infusions (DLI) has a prominent role in the management of disease recurrence. Although the highest remission rates are achieved in chronic myeloid leukaemia (CML), encouraging results have also been reported in chronic lymphoproliferative disorders. However, the experience of DLI in CML is not necessarily applicable to the management of lymphoproliferative diseases because of the heterogeneity of the conditioning regimens used in chronic lymphoid malignancies. We will review the role of DLI for different disease types in the context of conventional and reduced-intensity conditioning regimens. The factors influencing response and graft-versus-host disease as well as the optimal cell dose will be discussed. Finally, we will describe the main avenues currently being explored to improve the selectivity and efficacy of DLI. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17448964&query_hl=1 ER - TY - JFULL T1 - The anabolic effects of strontium increase the number of progenitor but not primitive haemopoietic stem cells A1 - Lymperi, S A1 - Horwood, N A1 - Cope, A A1 - Dazzi, F J1 - HAEMATOL-HEMATOL J Y1 - 2007/06// VL - 92 SN - 0390-6078 SP - 122 EP - 122 ER - TY - JFULL T1 - TCRzetadim lymphocytes define populations of circulating effector cells that migrate to inflamed tissues. A1 - Zhang, Z A1 - Gorman, CL A1 - Vermi, AC A1 - Monaco, C A1 - Foey, A A1 - Owen, S A1 - Amjadi, P A1 - Vallance, A A1 - McClinton, C A1 - Marelli-Berg, F A1 - Isomäki, P A1 - Russell, A A1 - Dazzi, F A1 - Vyse, TJ A1 - Brennan, FM A1 - Cope, AP J1 - Blood Y1 - 2007/05/15/ VL - 109 SN - 0006-4971 SP - 4328 EP - 4335 N2 - The T-cell receptor zeta (TCRzeta) chain is a master sensor and regulator of lymphocyte responses. Loss of TCRzeta expression has been documented in infectious, inflammatory, and malignant diseases, suggesting that it may serve to limit T-cell reactivity and effector responses at sites of tissue damage. These observations prompted us to explore the relationship between TCRzeta expression and effector function in T cells. We report here that TCRzeta(dim) lymphocytes are enriched for antigen-experienced cells refractory to TCR-induced proliferation. Compared to their TCRzeta(bright) counterparts, TCRzeta(dim) cells share characteristics of differentiated effector T cells but use accessory pathways for transducing signals for inflammatory cytokine gene expression and cell contact-dependent pathways to activate monocytes. TCRzeta(dim) T cells accumulate in inflamed tissues in vivo and have intrinsic migratory activity in vitro. Whilst blocking leukocyte trafficking with anti-TNF therapy in vivo is associated with the accumulation of TCRzeta(dim) T cells in peripheral blood, this T-cell subset retains the capacity to migrate in vitro. Taken together, the functional properties of TCRzeta(dim) T cells make them promising cellular targets for the treatment of chronic inflammatory disease. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17255353&query_hl=1 ER - TY - JFULL T1 - Low-intensity transplant regimens facilitate recruitment of donor-specific regulatory T cells that promote hematopoietic engraftment. A1 - Weng, L A1 - Dyson, J A1 - Dazzi, F J1 - Proc Natl Acad Sci U S A Y1 - 2007/05/15/ VL - 104 SN - 0027-8424 SP - 8415 EP - 8420 N2 - Low- or reduced-intensity conditioning regimens for allogeneic hemopoietic stem cell transplantation are effective at establishing donor hematopoietic engraftment and host-vs.-graft (HvG) tolerance. We investigated the mechanisms of HvG tolerance induction and maintenance in an animal model in which transplantation of sublethally irradiated female recipients with bone marrow (BM) from syngeneic male donors produces mixed chimerism. Splenocytes from chimeric mice inhibited HY-specific CD8(+) T cell responses both in vitro and in vivo, and their adoptive transfer facilitated donor hematopoietic engraftment. These properties were contained within the CD4(+)CD25(+) population. The conditioning protocol alone led to a proportional expansion of regulatory T cells (T(regs)), but the inhibitory activity was induced only if male BM was infused. The administration of anti-CD25-depleting antibodies to conditioned recipients at time of BM transplantation prevented donor-recipient chimerism but did not affect engraftment if performed after the establishment of chimerism, thus indicating that recipient T(regs) are required for the generation but not the maintenance of HvG tolerance. We conclude that donor-specific T(regs) of recipient origin are recruited when the donor antigens are present during reduced-intensity conditioning-induced T(reg) expansion. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17494756&query_hl=1 ER - TY - JFULL T1 - Proteomic analysis of proteins regulated by TRPS1 transcription factor in DU145 prostate cancer cells. A1 - Chang, GT A1 - Gamble, SC A1 - Jhamai, M A1 - Wait, R A1 - Bevan, CL A1 - Brinkmann, AO J1 - Biochim Biophys Acta Y1 - 2007/05// VL - 1774 SN - 0006-3002 SP - 575 EP - 582 N2 - The aim of the present study was to identify proteins differentially regulated by TRPS1 in human prostate cancer cells in order to better understand the role of TRPS1 in prostate cancer development. The proteomes of androgen-independent DU145 prostate cancer cells, that do not express TRPS1 and of genetically engineered DU145 cells that stable and inducible express recombinant TRPS1 protein, were compared. Using two-dimensional electrophoresis followed by mass spectrometric analysis, 13 proteins that were differentially expressed between these two cell lines were identified. These proteins represent a dominant reduction of expression of antioxidant proteins, including superoxide dismutase, protein disulfide isomerase A3 precursor, endoplasmin precursor and annexin A2. Furthermore, regulation was observed for mitochondrion-associated proteins, glycolytic enzymes, a cytoskeleton-associated protein, a nuclear protein and proteins involved in apoptosis. Our data indicate that overexpression of TRPS1 protein is correlated with reduced protein expression of certain antioxidants. This suggests a possible involvement of TRPS1 in oxidative stress, and possibly in apoptosis in androgen-independent DU145 prostate cancer cells. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17467349&query_hl=1 ER - TY - JFULL T1 - Response to donor lymphocyte infusions for chronic myeloid leukemia is dose-dependent: the importance of escalating the cell dose to maximize therapeutic efficacy. A1 - Simula, MP A1 - Marktel, S A1 - Fozza, C A1 - Kaeda, J A1 - Szydlo, RM A1 - Nadal, E A1 - Bua, M A1 - Rahemtulla, A A1 - Kanfer, E A1 - Marin, D A1 - Olavarria, E A1 - Goldman, JM A1 - Apperley, JF A1 - Dazzi, F J1 - Leukemia Y1 - 2007/05// VL - 21 SN - 0887-6924 SP - 943 EP - 948 N2 - Donor lymphocyte infusions (DLI) are an effective treatment for patients with chronic myeloid leukemia (CML) in relapse after allografting but the optimal cell dose has yet to be identified. To address this question, we investigated the factors affecting the dose required to achieve remission (effective cell dose, (ECD)) in 81 patients treated with an escalating dose regimen. The overall proportion of patients who achieved a molecular remission was 88%. The cumulative proportion of remitters increased significantly at each dose level. With a CD3(+) cell dose < or =10(7)/kg, 56% of patients in molecular/cytogenetic relapse obtained molecular remission, whereas only 20% of those in hematologic relapse did so. At the same cell dose, 58% of patients who received lymphocytes from volunteer unrelated donors achieved remission, as compared to 29% of those who received DLI from sibling donors. We conclude that the response to DLI is dose-dependent and that the ECD is influenced by the quantity and phase of CML at relapse and degree of donor/recipient histocompatibility. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17361226&query_hl=1 ER - TY - JFULL T1 - Metabolic modulation induced by chronic hypoxia in rats using a comparative proteomic analysis of skeletal muscle tissue. A1 - De Palma, S A1 - Ripamonti, M A1 - Vigano, A A1 - Moriggi, M A1 - Capitanio, D A1 - Samaja, M A1 - Milano, G A1 - Cerretelli, P A1 - Wait, R A1 - Gelfi, C J1 - J Proteome Res Y1 - 2007/05// VL - 6 SN - 1535-3893 SP - 1974 EP - 1984 N2 - Hypoxia-induced changes of rat skeletal muscle were investigated by two-dimensional difference in-gel electrophoresis (2D-DIGE) and mass spectrometry. The results indicated that proteins involved in the TCA cycle, ATP production, and electron transport are down-regulated, whereas glycolytic enzymes and deaminases involved in ATP and AMP production were up-regulated. Up-regulation of the hypoxia markers hypoxia inducible factor 1 (HIF-1alpha) and pyruvate dehydrogenase kinase 1 (PDK1) was also observed, suggesting that in vivo adaptation to hypoxia requires an active metabolic switch. The kinase protein, mammalian target of rapamycin (mTOR), which has been implicated in the regulation of protein synthesis in hypoxia, appears unchanged, suggesting that its activity, in this system, is not controlled by oxygen partial pressure. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17391017&query_hl=1 ER - TY - JFULL T1 - IL-10 induces IL-10 in primary human monocyte-derived macrophages via the transcription factor Stat3 A1 - Staples, KJ A1 - Smallie, T A1 - Williams, LM A1 - Foey, A A1 - Burke, B A1 - Foxwell, BMJ A1 - Ziegler-Heitbrock, L J1 - J IMMUNOL Y1 - 2007/04/15/ VL - 178 SN - 0022-1767 SP - 4779 EP - 4785 N2 - IL-10 is an important immunosuppressive cytokine that can down-regulate expression of other cytokines and has been shown to down-regulate itself. We show, in this study, that treatment of human monocyte-derived macrophages with IL-10 induces IL-10 mRNA in a dose- and time-dependent manner with an optimum induction at 100 ng/ml and at 6 h, whereas IL-10-induced IL-10 protein can be detected at 18 h. In the Same cells, IL-10 can partially suppress IL-10 mRNA induced by LPS, but only down to the level of IL-10-induced IL-10. An adenoviral luciferase reporter construct driven by the -195 IL-10 promoter, which contains a Stat motif, was readily induced by both IL-10 and LPS. Mutation of this Stat motif ablated IL-10 activation of this promoter, but not the LPS activation. Finally, we show that overexpression of a dominant-negative Stat3 protein will prevent IL-10 induction, but not LPS induction, of IL-10 mRNA. These data show that IL-10 induces IL-10 in monocyte-derived macrophages in an autocrine manner via activation of the transcription factor Stat3. ER - TY - JFULL T1 - IL-10 induces IL-10 in primary human monocyte-derived macrophages via the transcription factor Stat3. A1 - Staples, KJ A1 - Smallie, T A1 - Williams, LM A1 - Foey, A A1 - Burke, B A1 - Foxwell, BM A1 - Ziegler-Heitbrock, L J1 - J Immunol Y1 - 2007/04/15/ VL - 178 SN - 0022-1767 SP - 4779 EP - 4785 N2 - IL-10 is an important immunosuppressive cytokine that can down-regulate expression of other cytokines and has been shown to down-regulate itself. We show, in this study, that treatment of human monocyte-derived macrophages with IL-10 induces IL-10 mRNA in a dose- and time-dependent manner with an optimum induction at 100 ng/ml and at 6 h, whereas IL-10-induced IL-10 protein can be detected at 18 h. In the same cells, IL-10 can partially suppress IL-10 mRNA induced by LPS, but only down to the level of IL-10-induced IL-10. An adenoviral luciferase reporter construct driven by the -195 IL-10 promoter, which contains a Stat motif, was readily induced by both IL-10 and LPS. Mutation of this Stat motif ablated IL-10 activation of this promoter, but not the LPS activation. Finally, we show that overexpression of a dominant-negative Stat3 protein will prevent IL-10 induction, but not LPS induction, of IL-10 mRNA. These data show that IL-10 induces IL-10 in monocyte-derived macrophages in an autocrine manner via activation of the transcription factor Stat3. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17404258&query_hl=1 ER - TY - JFULL T1 - CD4+ CD25+ regulatory T-cells exhibit a higher T-cell receptor diversity than CD4+ CD25- conventional T-cells after allogeneic stem cell transplantation A1 - Fozza, C A1 - Nadal, E A1 - Longinotti, M A1 - Dazzi, F J1 - BONE MARROW TRANSPL Y1 - 2007/04// VL - 39 SN - 0268-3369 SP - S46 EP - S46 ER - TY - JFULL T1 - Inhibition of p38 mitogen-activated protein kinase is effective in the treatment of experimental crescentic glomerulonephritis and suppresses monocyte chemoattractant protein-1 but not IL-1beta or IL-6. A1 - Sheryanna, A A1 - Bhangal, G A1 - McDaid, J A1 - Smith, J A1 - Manning, A A1 - Foxwell, BM A1 - Feldmann, M A1 - Cook, HT A1 - Pusey, CD A1 - Tam, FW J1 - J Am Soc Nephrol Y1 - 2007/04// VL - 18 SN - 1046-6673 SP - 1167 EP - 1179 N2 - Activation of p38 mitogen-activated protein kinase (MAPK) is known to be important in cytokine production and cell survival in inflammation. This study examined the effect of inhibiting p38 MAPK after onset of renal injury in an experimental model of crescentic glomerulonephritis. Furthermore, this study investigated whether p38 MAPK inhibition would cause widespread suppression of the cytokine network in vivo or uncontrolled apoptosis. In the in vivo studies, daily treatment with a p38 MAPKalpha/beta inhibitor was started 1 h (early treatment study) or 4 d (late treatment study) after induction of nephrotoxic nephritis in Wistar Kyoto rats. The treated rats remained healthy with normal weight gain during the study. Both early and late treatment with p38 MAPK inhibitor reduced renal monocyte chemoattractant protein-1 (MCP-1) levels, the number of glomerular macrophages, the severity of tissue injury, and proteinuria compared with the vehicle group. Unexpected, treatment with p38 MAPK inhibitor did not suppress renal levels of IL-1beta or IL-6. In the in vitro study, the p38 MAPKalpha/beta inhibitor reduced production of MCP-1 and IL-6 by TNF-alpha-or IL-1beta-stimulated mesangial cells without any effect on cell viability or apoptosis. In conclusion, p38 MAPK inhibition is effective in reducing the severity of crescentic glomerulonephritis even when treatment is started after onset of disease. The therapeutic effect is associated with selective suppression of MCP-1, without widespread suppression of cytokine production or increased apoptosis. Therefore, p38 MAPK therapeutic blockade is a promising strategy in the treatment of antibody-mediated glomerulonephritis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17314328&query_hl=1 ER - TY - JFULL T1 - Proteome changes associated with hippocampal MRI abnormalities in the lithium pilocarpine-induced model of convulsive status epilepticus. A1 - Greene, ND A1 - Bamidele, A A1 - Choy, M A1 - de Castro, SC A1 - Wait, R A1 - Leung, KY A1 - Begum, S A1 - Gadian, DG A1 - Scott, RC A1 - Lythgoe, MF J1 - Proteomics Y1 - 2007/04// VL - 7 SN - 1615-9853 SP - 1336 EP - 1344 N2 - Convulsive status epilepticus is associated with subsequent hippocampal damage and development of mesial temporal sclerosis in a subset of individuals. The lithium pilocarpine model of status epilepticus (SE) in the rat provides a model in which to investigate the molecular and pathogenic process leading to hippocampal damage. In this study, a 2-DE-based approach was used to detect proteome changes in the hippocampus, at an early stage (2 days) after SE, when increased T2 values were detectable by magnetic resonance imaging. Gel image analysis was followed by LC-MS/MS identification of protein species that differed in abundance between pilocarpine-treated and control rats. The most significantly up-regulated species in the experimental animals was identified as heat shock 27-kDa protein, in line with findings in humans and in other experimental models of epilepsy. Additional up-regulated species included dihydropyrimidinase-related protein-2, cytoskeletal proteins (alpha-tubulin and ezrin) and dihydropteridine reductase. In summary, the hippocampus of rats subject to pilocarpine-induced SE exhibits specific changes in protein abundance, which likely relate to pathogenic, neuroprotective and neurogenic responses. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17366478&query_hl=1 ER - TY - JFULL T1 - The identification of surface proteins of Burkholderia pseudomallei. A1 - Harding, SV A1 - Sarkar-Tyson, M A1 - Smither, SJ A1 - Atkins, TP A1 - Oyston, PC A1 - Brown, KA A1 - Liu, Y A1 - Wait, R A1 - Titball, RW J1 - Vaccine Y1 - 2007/03/30/ VL - 25 SN - 0264-410X SP - 2664 EP - 2672 N2 - Burkholderia pseudomallei, the causative agent of the disease melioidosis is a human pathogen endemic in Northern Australia and South-East Asia. At present there is no available vaccine or effective treatment for this disease. Surface proteins play crucial roles in the host-pathogen interaction and have been exploited as vaccine candidates and diagnostic targets. Therefore, we wished to identify immunogenic surface proteins of B. pseudomallei. To this end we used two proteomic-based approaches in parallel: a biotinylation approach for the detection of surface located proteins identified 35 proteins, while screening with human sera identified 12 immunogenic proteins. Nine of these proteins were identified by both methods indicating that they may be both surface located and immunogenic: these proteins will be evaluated further as vaccine candidates and diagnostic targets. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17289218&query_hl=1 ER - TY - JFULL T1 - Attitudes to blood transfusion post arthroplasty surgery in the United Kingdom: A national survey. A1 - Young, SW A1 - Marsh, DJ A1 - Akhavani, MA A1 - Walker, CG A1 - Skinner, JA J1 - Int Orthop Y1 - 2007/03/30/ SN - 0341-2695 N2 - Five hundred orthopaedic surgeons and 336 anaesthetists were surveyed to assess current UK attitudes towards transfusion practice following arthroplasty surgery. Seventy-two percent of surgeons and 73% of anaesthetists responded to the survey. In an uncomplicated patient following total hip arthroplasty, 53.2% of surgeons and 63.1% of anaesthetists would transfuse at or below a haemoglobin (Hb) level of 8 g/dL. Surgeons tended to be more aggressive in their attitudes, with a mean transfusion threshold of 8.3 g/dL compared to 7.9 g/dL for anaesthetists (p < 0.01), and with 97% of surgeons transfusing two or more units compared to 78% of anaesthetists (p < 0.01). This threshold Hb increased if the patient was symptomatic (surgeons 9.3 g/dL, anaesthetists 8.8 g/dL, p < 0.05) or was known to have pre-existing ischaemic heart disease (surgeons 9.0 g/dL, anaesthetists 9.2 g/dL, p < 0.05). A wide variability in attitudes and practices is demonstrated, and the development and adoption of consensus guidelines needs to be encouraged if efforts to reduce the use of blood products are to succeed. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17396259&query_hl=1 ER - TY - JFULL T1 - Chest wall reconstruction using a turbocharged chimaeric anterolateral thigh flap. A1 - Gore, SM A1 - Akhavani, MA A1 - Kang, N A1 - Chana, JS J1 - J Plast Reconstr Aesthet Surg Y1 - 2007/03/26/ SN - 1748-6815 N2 - Extremely large chest wall defects may result following salvage oncological surgery. Typically these defects involve a large skin defect combined with a variable resected area of underlying muscle and ribs. In situations where the skin defect is very large the use of a large latissimus dorsi flap may require skin grafting to the donor site if a myocutaneous flap is used or to the recipient defect if a muscle-only flap is used. Alternatively a transverse rectus abdominis flap is a second option but in certain cases this may not be available. We describe the use of a free anterolateral thigh flap to reconstruct a chest wall defect and demonstrate the principle of side-to-side stacking of separate skin paddles to achieve skin closure of a massive defect whilst permitting primary closure of the donor site. The principle of turbocharging components of a chimaeric flap is also described. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17392046&query_hl=1 ER - TY - JFULL T1 - Alteration of the cortisol-cortisone shuttle in leprosy type 1 reactions in leprosy patients in Hyderabad, India. A1 - Andersson, AK A1 - Atkinson, SE A1 - Khanolkar-Young, S A1 - Chaduvula, M A1 - Jain, S A1 - Suneetha, L A1 - Suneetha, S A1 - Lockwood, DN J1 - Immunol Lett Y1 - 2007/03/15/ VL - 109 SN - 0165-2478 SP - 72 EP - 75 N2 - Regulation of inflammation in leprosy may be influenced by local concentrations of active cortisol and inactive cortisone, whose concentrations are regulated by enzymes in the cortisol-cortisone shuttle. We investigated the cortisol-cortisone shuttle enzymes in the skin of leprosy patients with type 1 reactions (T1R), which are characterised by skin and nerve inflammation. Gene expression of the shuttle enzymes were quantified in skin biopsies from 15 leprosy patients with new T1R before and during prednisolone treatment and compared with levels in skin biopsies from 10 borderline leprosy patients without reactions. Gene expression of 11beta-hydroxysteroid dehydrogenase (11beta-HSD) type 2, which converts cortisol to cortisone, is down-regulated in skin from T1R lesions. However expression levels of 11beta-HSD type 1, which converts cortisone to cortisol, were similar in skin with and without reactions and did not change during anti-leprosy drug treatment. Prednisolone treatment of patients with reactions is associated with an upregulation of 11beta-HSD2 expression in skin. The down regulation of 11beta-HSD2 at the beginning of a reaction may be caused by pro-inflammatory cytokines in the leprosy reactional lesion and may be a local attempt to down-regulate inflammation. However in leprosy reactions this local response is insufficient and exogenous steroids are required to control inflammation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17320974&query_hl=1 ER - TY - JFULL T1 - Expression of constitutively active STAT3 can replicate the cytokine-suppressive activity of interleukin-10 in human primary macrophages. A1 - Williams, LM A1 - Sarma, U A1 - Willets, K A1 - Smallie, T A1 - Brennan, F A1 - Foxwell, BM J1 - J Biol Chem Y1 - 2007/03/09/ VL - 282 SN - 0021-9258 SP - 6965 EP - 6975 N2 - There is general agreement that signal transducer and activation of transcription 3 (STAT3) is required to mediate the anti-inflammatory activities of interleukin (IL)-10. However, STAT3 is activated by multiple factors that do not share the anti-inflammatory activity of IL-10. The question remains whether STAT3 is sufficient for the anti-inflammatory effects or whether there are other signals required, as had been suggested previously. We set out to map the human IL-10 receptor and to identify the key elements involved in transducing the cytokine-suppressive effects of IL-10. We were able to show an absolute requirement for both of the tyrosine residues found within the YXXQ-STAT3-docking site within the IL-10 receptor 1 and that no other signals appeared to be required. We used a constitutively active STAT3 to determine whether expression of this factor could suppress lipopolysaccharide-induced tumor necrosis factor and IL-6 production. Our data show that STAT3 activity can suppress both IL-6 and tumor necrosis factor production in lipopolysaccharide-stimulated macrophages. However, in synovial fibroblasts, STAT3 did not suppress IL-6 production, suggesting that the cellular environment plays an important role in dictating whether STAT3 drives a pro- or anti-inflammatory response. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17194701&query_hl=1 ER - TY - JFULL T1 - Discovery of a vaccine antigen that protects mice from Chlamydia pneumoniae infection. A1 - Thorpe, C A1 - Edwards, L A1 - Snelgrove, R A1 - Finco, O A1 - Rae, A A1 - Grandi, G A1 - Guilio, R A1 - Hussell, T J1 - Vaccine Y1 - 2007/03/08/ VL - 25 SN - 0264-410X SP - 2252 EP - 2260 N2 - Chlamydiae are atypical intracellular bacteria that infect via mucosal surfaces causing, for example, trachoma, pneumonia, cervicitis, urethritis and infertility. Existing antibiotics are only partially effective and no vaccines are available. Using surface expressed or secreted proteins previously identified by genomics and proteomics we tested five as vaccines against intranasal challenge with Chlamydia pneumoniae. One antigen, LcrE, induced CD4+ and CD8+ T cell activation, type 1 cytokine secretion and neutralising antibodies and was completely effective in eliminating infection. Such antigens are highly conserved and essential to all Chlamydial species. The discovery of an effective vaccine for Chlamydiae pneumoniae has potential wide benefits for human health. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17275142&query_hl=1 ER - TY - JFULL T1 - Heat shock protein 27 functions in inflammatory gene expression and transforming growth factor-beta-activated kinase-1 (TAK1)-mediated signaling. A1 - Alford, KA A1 - Glennie, S A1 - Turrell, BR A1 - Rawlinson, L A1 - Saklatvala, J A1 - Dean, JL J1 - J Biol Chem Y1 - 2007/03/02/ VL - 282 SN - 0021-9258 SP - 6232 EP - 6241 N2 - Heat shock protein (HSP) 27 has long been known to be a component of the p38 mitogen-activated protein kinase (MAPK) signaling pathway. p38 MAPK has important functions in the inflammatory response, but the role of HSP27 in inflammation has remained unknown. We have used small interfering RNAs to suppress HSP27 expression in HeLa cells and fibroblasts and found that it is required for pro-inflammatory cell signaling and the expression of pro-inflammatory genes. HSP27 is needed for the activation by interleukin (IL)-1 of TAK1 and downstream signaling by p38 MAPK, JNK, and their activators (MKK-3, -4, -6, -7) and IKKbeta. IL-1-induced ERK activation appears to be independent of HSP27. HSP27 is required for both IL-1 and TNF-induced signaling pathways for which the most upstream common signaling protein is TAK1. HSP27 is also required for IL-1-induced expression of the pro-inflammatory mediators, cyclooxygenase-2, IL-6, and IL-8. HSP27 functions to drive cyclooxygenase-2 and IL-6 expression by augmenting the activation of the kinase downstream of p38 MAPK, MK2, resulting in stabilization of cyclooxygenase-2 and IL-6 mRNAs. The mechanism may not occur in cells of myeloid lineage because HSP27 protein was undetectable in human monocytes and murine macrophages. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17202147&query_hl=1 ER - TY - JFULL T1 - Galectin-1: a key effector of regulation mediated by CD4+CD25+ T cells. A1 - Garín, MI A1 - Chu, CC A1 - Golshayan, D A1 - Cernuda-Morollón, E A1 - Wait, R A1 - Lechler, RI J1 - Blood Y1 - 2007/03/01/ VL - 109 SN - 0006-4971 SP - 2058 EP - 2065 N2 - The naturally occurring population of dedicated regulatory T cells that coexpress CD4 and CD25 is known to play a key role in the maintenance of peripheral T-cell tolerance; however, their mechanism of action has remained obscure. Here we report that a member of the family of beta-galactoside-binding proteins, galectin-1, is overexpressed in regulatory T cells, and that expression is increased after activation. Most importantly, blockade of galectin-1 binding significantly reduced the inhibitory effects of human and mouse CD4+CD25+ T cells. Reduced regulatory activity was observed in CD4+CD25+ T cells obtained from galectin-1-homozygous null mutant mice. These results suggest that galectin-1 is a key effector of the regulation mediated by these cells. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17110462&query_hl=1 ER - TY - JFULL T1 - Adenoviral gene transfer of the endogenous inhibitor IkappaBalpha into human osteoarthritis synovial fibroblasts demonstrates that several matrix metalloproteinases and aggrecanases are nuclear factor-kappaB-dependent. A1 - Bondeson, J A1 - Lauder, S A1 - Wainwright, S A1 - Amos, N A1 - Evans, A A1 - Hughes, C A1 - Feldmann, M A1 - Caterson, B J1 - J Rheumatol Y1 - 2007/03// VL - 34 SN - 0315-162X SP - 523 EP - 533 N2 - OBJECTIVE: To investigate the role of the transcription factor nuclear factor-kB (NF-kappaB) in promoting inflammatory and destructive responses in human osteoarthritis (OA) synovial fibroblasts, by assessing the effect of NF-kappaB blockade on the production of cytokines and destructive enzymes. METHODS: Infection with adenoviruses transferring the beta-galactosidase gene was used to ascertain that the OA fibroblasts could be infected (> 95%). Using an adenovirus transferring the inhibitory subunit IkappaBa, effective inhibition of NF-kappaB was achieved. The expression and production of several pro- and antiinflammatory cytokines and mediators, the major matrix metalloproteinases (MMP 1, 3, and 13), their main inhibitor tissue inhibitor of metalloproteinase-1 (TIMP-1), and the aggrecanases (ADAMTS4 and ADAMTS5) were measured by ELISA and/or reverse transcription-polymerase chain reaction, and their dependence on NF-kappaB evaluated. RESULTS: The production of interleukin 6 (IL-6), monocyte chemoattractant protein-1, and RANTES was potently inhibited by IkBa overexpression, irrespective of stimulus, but IL-8 was unaffected. The p55 soluble tumor necrosis factor (TNF) receptor was unaffected, but the p75 soluble TNF receptor was potently inhibited by IkBa overexpression. MMP-1, MMP-3, and MMP-13 were inhibited by IkappaBa overexpression, at both the mRNA and protein levels, whereas TIMP-1 was unaffected. The mRNA gene expression of ADAMTS4 was also inhibited by IkappaBa overexpression, particularly in IL-1-stimulated cells, but ADAMTS5 was unaffected. CONCLUSION: In OA synovial fibroblasts, inhibition of NF-kappaB has a beneficial effect on the balance between the expression of proinflammatory cytokines and antiinflammatory mediators. Inhibition of this transcription factor also results in the decreased expression of several destructive metalloproteinases and also the ADAMTS4 aggrecanase. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17295438&query_hl=1 ER - TY - JFULL T1 - Increased nitric oxide production regulates T cell function in rheumatoid arthritis A1 - Nagy, G A1 - Clark, JM A1 - Buzas, E A1 - Gorman, C A1 - Falus, A A1 - Cope, AP J1 - ANN RHEUM DIS Y1 - 2007/03/01/ VL - 66 SN - 0003-4967 SP - A69 EP - A69 ER - TY - JFULL T1 - Increased frequencies of CD4(+)CD25(high) T(regs) correlate with disease relapse after allogeneic stem cell transplantation for chronic myeloid leukemia. A1 - Nadal, E A1 - Garin, M A1 - Kaeda, J A1 - Apperley, J A1 - Lechler, R A1 - Dazzi, F J1 - Leukemia Y1 - 2007/03// VL - 21 SN - 0887-6924 SP - 472 EP - 479 N2 - The therapeutic efficacy of allogeneic hemopoietic stem cell transplantation (SCT) for chronic myeloid leukemia (CML) largely relies on the graft-versus-leukemia (GvL) effect exerted by donor T cells. CD4(+)CD25(high) regulatory T cells (T(regs)) have been shown to downregulate antitumor responses but their role on GvL has not been evaluated. We performed a cross-sectional study in which we enumerated and characterized CD4(+)CD25(high) T(regs) in the peripheral blood of CML patients undergoing allogeneic SCT. We documented higher frequencies of T(regs) in patients after transplant as compared to normal controls and newly diagnosed patients. The increment was particularly evident in patients who had received their SCT 18 months before. In vitro functional studies demonstrated that the T(regs) purified from SCT patients exhibited a more potent suppressive activity than T(regs) isolated from healthy volunteers. Patients in whom T(regs) numbers were higher than controls more than 18 months after SCT showed evidence of disease relapse. Although the increment in T(regs) might have an advantageous effect on graft rejection in the early phase post-transplant, our data suggest that T(regs) exert an inhibitory effect on GvL. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17215853&query_hl=1 ER - TY - JFULL T1 - TCR-zeta dim T cells define a population of circulating effector cells that migrate to inflamed tissues A1 - Gorman, CL A1 - Zhang, Z A1 - Vermi, AC A1 - Monaco, C A1 - Marelli-Berg, F A1 - Dazzi, F A1 - Cope, AP J1 - ANN RHEUM DIS Y1 - 2007/03/01/ VL - 66 SN - 0003-4967 SP - A17 EP - A17 ER - TY - JFULL T1 - Characterising pre and post-mortem factors influencing human post-mortem brain proteomic studies A1 - Cotter, D A1 - English, J A1 - Dicker, P A1 - Wait, R A1 - Dunn, M J1 - SCHIZOPHRENIA BULL Y1 - 2007/03// VL - 33 SN - 0586-7614 SP - 258 EP - 258 ER - TY - JFULL T1 - Synovial fibroblasts are important mediators of synovial angiogenesis in the hypoxic rheumatoid joint A1 - Larsen, H A1 - Feldmann, M A1 - Paleolog, E J1 - ANN RHEUM DIS Y1 - 2007/03/01/ VL - 66 SN - 0003-4967 SP - A61 EP - A61 ER - TY - JFULL T1 - Innate and adaptive immune interactions during respiratory infection A1 - Hussell, T J1 - IMMUNOLOGY Y1 - 2007/03// VL - 120 SN - 0019-2805 SP - 24 EP - 24 ER - TY - JFULL T1 - QUIPP: A novel tyrosine phosphoprotein implicated in toll like receptor signalling A1 - Peirce, MJ A1 - Testar, J A1 - Brook, M A1 - Begum, S A1 - Wait, R A1 - Hussell, T A1 - Cope, AP J1 - ANN RHEUM DIS Y1 - 2007/03/01/ VL - 66 SN - 0003-4967 SP - A17 EP - A17 ER - TY - JFULL T1 - Analysis of skin cancer risk factors in immunosuppressed renal transplant patients shows high levels of UV-specific tandem CC to TT mutations of the p53 gene. A1 - Queille, S A1 - Luron, L A1 - Spatz, A A1 - Avril, MF A1 - Ribrag, V A1 - Duvillard, P A1 - Hiesse, C A1 - Sarasin, A A1 - Armand, JP A1 - Daya-Grosjean, L J1 - Carcinogenesis Y1 - 2007/03// VL - 28 SN - 0143-3334 SP - 724 EP - 731 N2 - Immunosuppressed renal transplant recipients (RTRs) are predisposed to non-melanoma skin cancers (NMSCs), predominantly squamous cell carcinomas (SCCs). We have analyzed skin lesions from RTRs with aggressive tumors for p53 gene modifications, the presence of Human Papillomas Virus (HPV) DNA in relation to the p53 codon 72 genotype and polymorphisms of the XPD repair gene. We detected 24 p53 mutations in 15/25 (60%) NMSCs, 1 deletion and 23 base substitutions, the majority (78%) being UV-specific C to T transitions at bipyrimidine sites. Importantly, 35% (6/17) are tandem mutations, including 4 UV signature CC to TT transitions possibly linked to modulated DNA repair caused by the immunosuppressive drug cyclosporin A (CsA). We found 8 p53 mutations in 7/17 (41%) precancerous actinic keratosis (AK), suggesting that p53 mutations are early events in RTR skin carcinogenesis. Immunohistochemical analysis shows a good correlation between p53 accumulation and mutations. HPV DNA was detected in 78% of skin lesions (60% Basal Cell Carcinomas, 82%AK and 79% SCCs). Thus, immunosuppression has increased the risk of infections by HPVs, predominantly epidermodysplasia verruciformis, speculated to play a role in skin cancer development. No association is found between HPV status and p53 mutation. Moreover, p53 codon 72 or frequencies of three XPD genotypes of RTRs are comparable with control populations. The p53 mutation spectrum, presenting a high level of CC to TT mutations, shows that the UV component of sunlight is the major risk factor and modulated DNA repair by immunosuppressive drug treatment may be significant in the skin carcinogenesis of RTRs. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17065198&query_hl=1 ER - TY - JFULL T1 - Upregulation of OX40 and OX40 ligand during respiratory virus infection A1 - Cavanagh, MM A1 - Gwyer, E A1 - Snelgrove, RJ A1 - Hussell, T J1 - IMMUNOLOGY Y1 - 2007/03// VL - 120 SN - 0019-2805 SP - 27 EP - 27 ER - TY - JFULL T1 - Factors for graft-versus-host disease after donor lymphocyte infusions with an escalating dose regimen: lack of association with cell dose. A1 - Fozza, C A1 - Szydlo, RM A1 - Abdel-Rehim, MM A1 - Nadal, E A1 - Goldman, JM A1 - Apperley, JF A1 - Dazzi, F J1 - Br J Haematol Y1 - 2007/03// VL - 136 SN - 0007-1048 SP - 833 EP - 836 N2 - We investigated the risk factors for graft-versus-host disease (GVHD) in 82 patients treated with donor lymphocyte infusions (DLI) using an escalating dose regimen for chronic myeloid leukaemia in relapse following conventional allografting. Two factors emerged as predictors of both acute and chronic GVHD: the infusion of male recipients with lymphocytes from a female donor and the interval between transplant and last DLI, but only the first remained significant at multivariate analysis. Surprisingly, lymphocyte dose did not influence the incidence of GVHD. Our results suggest that DLI can be given in large cell doses without increasing the risk of GVHD. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17341269&query_hl=1 ER - TY - JFULL T1 - Increased nitric oxide production regulates T cell function in rheumatoid arthritis A1 - Nagy, G A1 - Clark, JM A1 - Buzas, E A1 - Gorman, C A1 - Falus, A A1 - Cope, AP J1 - JOINT BONE SPINE Y1 - 2007/03// VL - 74 SN - 1297-319X SP - 215 EP - 216 ER - TY - JFULL T1 - Circulating endothelial progenitor cells as a link between synovial vascularity and cardiovascular mortality in rheumatoid arthritis. A1 - Akhavani, MA A1 - Larsen, H A1 - Paleolog, E J1 - Scand J Rheumatol Y1 - 2007/03// VL - 36 SN - 0300-9742 SP - 83 EP - 90 N2 - Cardiovascular disease refers to the class of diseases that involve the heart and/or blood vessels (arteries and veins). Most Western countries face high and ever-increasing rates of cardiovascular disease. Each year, more Americans are killed by heart disease than by cancer. Diseases of the heart alone cause 30% of all deaths, with other diseases of the cardiovascular system causing substantial further deaths and disability. Indeed, cardiovascular disease is the major cause of death and disability in the USA and most European countries. The development of the vascular systems requires an intricate interplay of molecules such as vascular endothelial growth factor and endothelial progenitor cells. A defective vascular repair/regeneration is thought to be responsible for propagation of atherosclerosis, a key feature of cardiovascular disease. This is partly attributed to a reduction in the circulating endothelial progenitor cells in peripheral blood. Patients with rheumatoid arthritis (RA) have a higher than average incidence of cardiovascular disease in comparison with the general population, with an increased risk of stroke and myocardial infarction, and an increased risk of fatality following myocardial infarction. This review focuses on the current evidence linking the role played by endothelial progenitor cells to the development of cardiovascular disease and why this might relate to the increased risk observed in RA patients. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17476612&query_hl=1 ER - TY - JFULL T1 - Proteomic analysis of white matter from the dorsolateral prefrontal cortex in schizophrenia demonstrated alterations in synaptic associated proteins A1 - English, J A1 - Dicker, P A1 - Wait, R A1 - O'Donoghue, N A1 - Pennington, S A1 - Dunn, M A1 - Cotter, D J1 - SCHIZOPHRENIA BULL Y1 - 2007/03// VL - 33 SN - 0586-7614 SP - 259 EP - 260 ER - TY - JFULL T1 - Long-term alteration of innate immunity at mucosal surfaces after recovery from viral infection A1 - Didierlaurent, A A1 - Snelgrove, R A1 - Low, L A1 - Sirard, JC A1 - Hussell, T J1 - IMMUNOLOGY Y1 - 2007/03// VL - 120 SN - 0019-2805 SP - 25 EP - 25 ER - TY - JFULL T1 - Growth and protein profile changes in Lepidium sativum L. plantlets exposed to cadmium A1 - Gianazza, E A1 - Wait, R A1 - Sozzi, A A1 - Regondi, S A1 - Saco, D A1 - Labrac, M A1 - Agradi, E J1 - ENVIRON EXP BOT Y1 - 2007/03// VL - 59 SN - 0098-8472 SP - 179 EP - 187 N2 - Plant metabolic response to heavy metal stress is still to be clarified. The present investigation was undertaken to examine the influence of different concentrations of cadmium on the Lepidium sativum L. plantlets. Exposure of seedlings of L. sativum L. to increasing concentrations of cadmium results in the growth inhibition and in the accumulation of proteins in the 10-25 kDa range in cotyledons and hypocotyls of the plantlets. Most of these proteins are also found in extracts of L. sativum seeds. Analysis by ESI-MS after two-dimensional electrophoresis showed that these proteins exhibit sequences similar to those of storage proteins from various Cruciferae species. The response to metal exposure during germination and initial plantlet elongation thus involves inhibition of both storage protein catabolism and plant protein anabolism. In addition, two of the proteins present in higher amounts in plantlets exposed to cadmium heat-shock, in agreement with literature data, and jasmonate-like inducible protein are related to cellular stress and another two (LEAs or late embryogenesis abundant) are involved in embryogenesis. Changes in protein expression can be detected by two-dimensional electrophoresis after exposure to heavy metal concentrations lower than those at which morphometric changes become evident. Proteomics of germinating L. sativiun thus constitutes a very sensitive too] for evaluating environmental pollution. (c) 2006 Elsevier B.V. All rights reserved. ER - TY - JFULL T1 - Identification of target mRNAs regulated by both p38 MAPK and tristetraprolin in murine macrophages A1 - Tudor, C A1 - Hitti, E A1 - Blackshear, PJ A1 - Clark, AR A1 - Saklatvala, J A1 - Dean, JLE J1 - IMMUNOLOGY Y1 - 2007/03// VL - 120 SN - 0019-2805 SP - 16 EP - 16 ER - TY - JFULL T1 - Anti-CD3 antibody-induced suppression of collagen-induced arthritis in the DBA/1 mouse is associated with expansion of regulatory T cell subsets A1 - Notley, CA A1 - Criado, G A1 - Inglis, JJ A1 - Cope, A A1 - Williams, RO J1 - IMMUNOLOGY Y1 - 2007/03// VL - 120 SN - 0019-2805 SP - 7 EP - 8 ER - TY - JFULL T1 - Selective use of TRAM in lipopolysaccharide (LPS) and lipoteichoic acid (LTA) induced NF-kappaB activation and cytokine production in primary human cells: TRAM is an adaptor for LPS and LTA signaling. A1 - Sacre, SM A1 - Lundberg, AM A1 - Andreakos, E A1 - Taylor, C A1 - Feldmann, M A1 - Foxwell, BM J1 - J Immunol Y1 - 2007/02/15/ VL - 178 SN - 0022-1767 SP - 2148 EP - 2154 N2 - TLR signal via Toll-IL-1R (TIR) homology domain-containing adaptor proteins. One of these adaptors, Toll-IL-1R domain-containing adaptor inducing IFN-beta-related adaptor molecule (TRAM), has been shown to be essential for TLR4 signaling in TRAM(-/-) mice and cell lines. Previously, we showed that MyD88 or Mal dominant-negative constructs did not inhibit LPS induction of cytokines in primary human M-CSF-derived macrophages. A possible explanation was redundancy of the adaptors during LPS signaling. TRAM is a suitable candidate to compensate for these adaptors. To investigate a potential role for TRAM in LPS signaling in human M-CSF-derived macrophages, we engineered an adenoviral construct expressing dominant-negative TRAM-C117H (AdTRAMdn). Synovial fibroblasts (SF) and human umbilical endothelial cells (HUVECs) were used as a nonmyeloid comparison. AdTRAMdn inhibited LPS-induced signaling in SFs and HUVECs, reducing NF-kappaB activation and cytokine production, but did not inhibit LPS signaling in M-CSF-derived human macrophages. Further investigation of other TLR ligands showed that AdTRAMdn was also able to inhibit signaling initiated by lipoteichoic acid, a TLR2 ligand, in SFs and HUVECs and lipoteichoic acid and macrophage-activating lipopeptide 2 signaling was also inhibited in TRAM(-/-) murine embryonic fibroblasts. We conclude that TRAM is an adaptor protein for both TLR4 and TLR2/6 signaling in SFs, HUVECs, and murine embryonic fibroblasts, but cannot demonstrate a role in human macrophages. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17277119&query_hl=1 ER - TY - JFULL T1 - T-cell receptor repertoire usage after allografting differs between CD4+CD25+ regulatory T cells and their CD4+CD25- counterpart. A1 - Fozza, C A1 - Nadal, E A1 - Longinotti, M A1 - Dazzi, F J1 - Haematologica Y1 - 2007/02// VL - 92 SN - 1592-8721 SP - 206 EP - 214 N2 - BACKGROUND AND OBJECTIVES: After allogeneic haematopoietic stem cell transplantation (SCT) the whole T-cell receptor (TCR) repertoire shows a markedly skewed pattern for 2-3 years. A small fraction of CD4+ T cells is represented by CD25+ regulatory lymphocytes (Treg), which play a crucial role in modulating peripheral tolerance. To investigate their ability to react to the massive antigenic stimulation generated in an allogeneic host, which could significantly affect their pattern of reconstitution, we analyzed the TCR repertoire of Treg after SCT, focusing on the degree of similarity to CD4+CD25- conventional T cells (Tconv). DESIGN AND METHODS: We assessed the TCR Vbeta repertoire of Treg in ten patients who had received allogeneic SCT, by using complementarity determining region 3 (CDR3) spectratyping. We developed a new similarity score for the analysis. This score expresses the proportion of Vbeta with similar profile between Treg and Tconv. RESULTS: For up to 3 years after SCT the repertoires of Treg and Tconv were characterized by several Vbeta with different profiles between the two cell subsets, while they were extremely similar in patients more than 3 years post-allografting (similarity score= 0.90 vs. 0.61). The differences observed early after SCT were mainly ascribable to Vbeta expressing an oligoclonal profile in Tconv but not in Treg. INTERPRETATION AND CONCLUSIONS: Our data show that the TCR repertoires of Treg and Tconv are significantly different early post-SCT, while they tend to become identical with full reconstitution. This difference could reflect either a discrepancy in the in vivo reactivity against common antigenic stimulations or be the result of different post-transplant ontogeny. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17296570&query_hl=1 ER - TY - JFULL T1 - Mapping the 5-50-kDa fraction of human amniotic fluid proteins by 2-DE and ESI-MS A1 - Gianazza, E A1 - Wait, R A1 - Begum, S A1 - Eberini, I A1 - Campagnoli, M A1 - Labo, S A1 - Galliano, M J1 - PROTEOM CLIN APPL Y1 - 2007/02// VL - 1 SP - 167 EP - 175 N2 - This report presents a proteomic analysis and provides a reference map of the 5-50-kDa components of normal amniotic fluid collected in gestational weeks 16-18. Early amniocentesis samples were pooled and proteins with molecular mass lower than albumin were separated by gel filtration chromatography. The 2-DE protocol was optimized for the separation of the small proteins and peptides in the fraction of interest. A total of 132 Coomassie blue-stained protein spots were analyzed, following in-gel tryptic digestion, by ESI-MS/MS and 49 different gene products were identified. The treatment with alkaline phosphatase caused the shift of the phosphoisoforms of insulin-like growth factor-binding protein-1 and of the N-terminal osteopontin fragment. Of the 33 full-length proteins identified in the 2-DE profile, 23 had not been previously detected in the amniotic fluid and, of these, 22 are not present in the human plasma proteome under physiological conditions. Fragments of 16 larger proteins were identified and the sequence coverage data revealed that several correspond to autonomous domains that may have biological roles on their own. Several of the detected proteins and peptides appear to be involved in critical regulatory processes associated with placentation and early development, thus representing potential markers of various physiological or pathological conditions. ER - TY - JFULL T1 - Role of calcineurin in the regulation of human lung mast cell and basophil function by cyclosporine and FK506. A1 - Harrison, CA A1 - Bastan, R A1 - Peirce, MJ A1 - Munday, MR A1 - Peachell, PT J1 - Br J Pharmacol Y1 - 2007/02// VL - 150 SN - 0007-1188 SP - 509 EP - 518 N2 - BACKGROUND AND PURPOSE: Cyclosporine and FK506 are thought to act by targeting the Ca2+-dependent protein phosphatase, calcineurin. The aim of the present study was to determine whether cyclosporine and FK506 stabilize mast cells and basophils by interacting with calcineurin. EXPERIMENTAL APPROACH: The effects of cyclosporine and FK506 on the IgE-mediated release of histamine from mast cells and basophils were evaluated. The presence of calcineurin in cells was determined by Western blotting. Ca2+-dependent protein phosphatase activities were assessed in cell extracts using a synthetic phosphorylated peptide that is known to serve as a substrate for calcineurin. KEY RESULTS: FK506 was about 100-fold more potent than cyclosporine as an inhibitor of IgE-dependent histamine release from mast cells and basophils. Immunoblotting of solubilized preparations of purified cells demonstrated the presence of calcineurin in mast cells and basophils. In enzyme assays, mast cells expressed approximately 7-fold higher Ca2+-dependent protein phosphatase activity than basophils. Whereas cyclosporine effectively inhibited Ca2+-dependent protein phosphatase activity in cell extracts, FK506 was considerably less effective. CONCLUSIONS AND IMPLICATIONS: FK506 and cyclosporine inhibit the stimulated release of histamine from mast cells and basophils. However, the ability of cyclosporine, but not FK506, to inhibit Ca2+-dependent protein phosphatase activity questions whether FK506 stabilizes mast cells and basophils by interacting with calcineurin. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17200674&query_hl=1 ER - TY - JFULL T1 - Etanercept plus sulfasalazine: added value in rheumatoid arthritis? A1 - Fisher, BA A1 - Taylor, PC J1 - Nat Clin Pract Rheumatol Y1 - 2007/02// VL - 3 SN - 1745-8382 SP - 70 EP - 71 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17228310&query_hl=1 ER - TY - JFULL T1 - Predicting the outcome of surgery for the proximal interphalangeal joint in Dupuytren's disease. A1 - Misra, A A1 - Jain, A A1 - Ghazanfar, R A1 - Johnston, T A1 - Nanchahal, J J1 - J Hand Surg [Am] Y1 - 2007/02// VL - 32 SN - 0363-5023 SP - 240 EP - 245 N2 - PURPOSE: We prospectively studied the outcome of limited Dupuytren's fasciectomy, in combination with joint release if necessary, for disease involving 49 proximal interphalangeal joints (PIPJs) to identify factors that predispose to recurrent PIPJ contracture. METHODS: Thirty-seven patients were treated over a 4-year period. The flexion contracture of the PIPJ was measured before surgery, immediately after surgery, and at more than 1 year after surgery. RESULTS: A mean preoperative flexion contracture of 67 degrees +/- 22 degrees was corrected to 6 degrees +/- 10 degrees at the time of surgery and 25 degrees +/- 25 degrees at the follow-up evaluation. There was a positive correlation between the severity of the preoperative flexion contracture and recurrent deformity, with a preoperative contracture greater than 60 degrees leading to significantly worse outcome. Incomplete correction of PIPJ flexion contracture during surgery and poor postoperative compliance with therapy were also associated with worse recurrent joint contractures. The digit involved and the necessity for joint release did not significantly affect outcome. CONCLUSIONS: In the absence of recurrent Dupuytren's disease, severe preoperative deformity, incomplete correction at surgery, and noncompliance with therapy predispose patients to worse PIPJ contracture. TYPE OF STUDY/LEVEL OF EVIDENCE: Prognostic II. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17275601&query_hl=1 ER - TY - JFULL T1 - Mesenchymal stem cells inhibit proliferation and apoptosis of tumor cells: impact on in vivo tumor growth. A1 - Ramasamy, R A1 - Lam, EW A1 - Soeiro, I A1 - Tisato, V A1 - Bonnet, D A1 - Dazzi, F J1 - Leukemia Y1 - 2007/02// VL - 21 SN - 0887-6924 SP - 304 EP - 310 N2 - Mesenchymal stem cells (MSC) have received much attention in the field of hematopoietic stem cell transplantation because not only do they support hematopoiesis but also exhibit a profound immunosuppressive activity that can be exploited to prevent undesired alloreactivity. We have previously shown that their immunosuppressive activity is mainly exerted at the level of T-cell proliferation. Here, we show that MSC exhibit a similar antiproliferative activity on tumor cells of hematopoietic and non hematopoietic origin. In vitro, MSC produced the transient arrest of tumor cells in the G(1) phase of cell cycle; this was accompanied by a reduction in the apoptotic rate even when survival factors were limiting. However, when tumor cells were injected into non-obese diabetic-severe combined immunodeficient mice in conjunction with MSC, their growth was much faster as compared to the group receiving only tumor cells. To explain the discrepancy between the in vitro and in vivo behavior, we suggest that MSC have the ability to form a cancer stem cell niche in which tumor cells can preserve the potential to proliferate and sustain the malignant process. We conclude that the clinical use of MSC in conditions in which a malignant disease is involved should be handled with extreme caution. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17170725&query_hl=1 ER - TY - JFULL T1 - Chest wall reconstruction using a prolene/hydroxyapatite paste sandwich. A1 - Akhavani, MA A1 - Gore, S A1 - Kang, N J1 - Plast Reconstr Surg Y1 - 2007/02// VL - 119 SN - 1529-4242 SP - 761 EP - 762 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17230134&query_hl=1 ER - TY - JFULL T1 - Management of severe open ankle injuries. A1 - Khan, U A1 - Smitham, P A1 - Pearse, M A1 - Nanchahal, J J1 - Plast Reconstr Surg Y1 - 2007/02// VL - 119 SN - 1529-4242 SP - 578 EP - 589 N2 - BACKGROUND: Functional outcome after reconstruction of open ankle injuries has not been well presented in the literature. The authors present the functional results of 24 patients who sustained complex ankle injuries. METHODS: Patients were assessed using three scoring systems (a modified A/O score, the Enneking score, and the AOFAS) and subdivided into two groups: those primarily treated at Charing Cross Hospital according to strict protocols combining orthopedic and plastic surgical techniques (group P) and those secondarily treated who were transferred to Charing Cross Hospital after initial management at a remote unit (group S). RESULTS: There were nine patients (37.5 percent) in group P and 15 (62.5 percent) in group S. Eighteen patients (75 percent) underwent free-tissue transfer. Sixteen patients (67 percent) were assessed (group P, n = 7; group S, n = 9) for return of function using the Enneking score. Mean time to assessment was 10.5 months for group P and 11.4 months for group S. Mean Enneking percentage score was 75 for group P and 72.2 for group S. There were no significant differences (p > 0.05) between these scores. The mean time to union was 19 weeks (n = 5) for group P and 24 weeks (n = 7) for group S. The mean AOFAS Ankle-Hindfoot Scores were comparable to the Enneking scores when independent observers undertook this assessment. Most patients in both groups reported difficulty with descent of stairs. CONCLUSIONS: Although the authors were able to achieve a similar return of function for both groups, group S patients needed at least one more operation. In cases of ankle fracture where there is significant soft-tissue injury (either closed or open), representing a complex injury, the authors recommend making no attempt to internally fix the fracture and instead referring the patient to a specialist center for combined orthoplastic attention. If this is not immediately at hand, screw fixation of the medial malleolus should be undertaken after open reduction. The lateral malleolus should not be internally fixed, but should it require control, external fixation is the preferred method of skeletal stabilization. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17230094&query_hl=1 ER - TY - JFULL T1 - Mesenchymal stem cells are effective at preventing but not at treating GvJD A1 - Tisato, V A1 - Naresh, K A1 - Navarrete, C A1 - Dazzi, F J1 - BIOL BLOOD MARROW TR Y1 - 2007/02// VL - 13 SN - 1083-8791 SP - 44 EP - 45 ER - TY - JFULL T1 - The Toll-like receptor adaptor proteins MyD88 and Mal/TIRAP contribute to the inflammatory and destructive processes in a human model of rheumatoid arthritis. A1 - Sacre, SM A1 - Andreakos, E A1 - Kiriakidis, S A1 - Amjadi, P A1 - Lundberg, A A1 - Giddins, G A1 - Feldmann, M A1 - Brennan, F A1 - Foxwell, BM J1 - Am J Pathol Y1 - 2007/02// VL - 170 SN - 0002-9440 SP - 518 EP - 525 N2 - The widespread distribution of Toll-like receptors (TLRs) and their ligands raises the question whether they contribute to the production of inflammatory and tissue destructive molecules in rheumatoid arthritis (RA). We examined the expression and function of TLR2 and TLR4 and their downstream signaling adaptors MyD88 and Mal/TIRAP in synovial membrane cultures from RA tissue. Both TLR2 and TLR4 were detected by flow cytometry, and stimulation with TLR2 and TLR4 ligands augmented the spontaneous production of tumor necrosis factor-alpha, interleukin (IL)-6, and IL-8, indicating that TLR2 and TLR4 are functional in these cultures. In addition, overexpression of dominant-negative forms of MyD88 and Mal/TIRAP significantly down-regulated the spontaneous production of cytokines tumor necrosis factor-alpha, IL-6, and vascular endothelial growth factor, and enzymes MMP-1, MMP-2, MMP-3, and MMP-13 in RA synovial membrane cell cultures. Because TLR2 and TLR4 require both MyD88 and Mal/TIRAP for signaling, this study suggests that TLR function may regulate the expression of these factors in the RA synovium. Conditioned media from synovial membrane cell cultures stimulated human macrophages in a MyD88- and Mal-dependent manner, suggesting the release of a TLR ligand(s) from these cells. Thus, TLRs not only protect against infection but may also promote the inflammatory and destructive process in RA. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17255320&query_hl=1 ER - TY - JFULL T1 - Reduced intensity conditioning allografting induces the generation of antigen-specific regulatory T cells necessary for graft tolerance A1 - Weng, L A1 - Dyson, J A1 - Dazzi, F J1 - BIOL BLOOD MARROW TR Y1 - 2007/02// VL - 13 SN - 1083-8791 SP - 104 EP - 104 ER - TY - JFULL T1 - Inflammatory signaling in cartilage: MAPK and NF-kappaB pathways in chondrocytes and the use of inhibitors for research into pathogenesis and therapy of osteoarthritis. A1 - Saklatvala, J J1 - Curr Drug Targets Y1 - 2007/02// VL - 8 SN - 1873-5592 SP - 305 EP - 313 N2 - Osteoarthritis is characterised by degeneration of articular cartilage. It is thought to be primarily a disease of cartilage. Inflammatory response genes, such as proteinases, cyclooxygenase, and cytokines are implicated in its pathogenesis. The evidence for expression of these genes in articular cartilage in osteoarthritis is reviewed. The expression of inflammatory response genes is controlled by four major intracellular signalling pathways. These lead to activation of the three mitogen-activated protein kinases (MAPK) and the transcriptional regulator nuclear factor kappa (NFkappa)-B. The current state of knowledge of the structure of these pathways is summarized. Pharmacological inhibitors of the protein kinases of the pathways in current use are described, and insights into chondrocyte gene expression obtained with them are discussed. Very limited use of these inhibitors has yet been made in animal models of osteoarthritis. The main use of the inhibitors in the near future will be in investigation of pathogenetic mechanisms in osteoarthritis, both in experimental animals and in vitro, with a view to identifying therapeutic targets. Prospects for using signalling pathway inhibitors for therapy in osteoarthritis are distant. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17305508&query_hl=1 ER - TY - JFULL T1 - Increased CD4+CD25high+regulatory T-cell are associated with disease relapse after allogeneic stem cell transplantation (SCT) for chronic myeloid leukaemia (CML) A1 - Nadal, E A1 - Kaeda, J A1 - Apperley, JF A1 - Lechler, R A1 - Dazzi, F J1 - BIOL BLOOD MARROW TR Y1 - 2007/02// VL - 13 SN - 1083-8791 SP - 15 EP - 15 ER - TY - JFULL T1 - Mesenchymal stem cells inhibit dendritic cell differentiation and function by preventing entry into the cell cycle. A1 - Ramasamy, R A1 - Fazekasova, H A1 - Lam, EW A1 - Soeiro, I A1 - Lombardi, G A1 - Dazzi, F J1 - Transplantation Y1 - 2007/01/15/ VL - 83 SN - 0041-1337 SP - 71 EP - 76 N2 - BACKGROUND: Mesenchymal stem cells (MSCs) play a crucial role in hematopoietic development and have been shown to exert a powerful immunosuppressive effect. In this study, we investigated the effect of bone marrow MSC on the differentiation and function of peripheral blood monocytes into dendritic cells (DCs). METHODS: Human MSCs, generated from normal bone marrow, were added to peripheral blood monocytes stimulated in vitro with granulocyte-macrophage colony stimulating factor and interleukin-4 to become DCs. Monocytes were then examined for the expression of markers characteristic of DCs and their ability to stimulate allogeneic T cells. In addition, the effect of MSCs on the cell cycle of monocyte-derived DCs and the expression of various cell cycle proteins were analyzed by cytometric analysis and Western blotting with specific antibodies. RESULTS: MSCs blocked the differentiation of monocytes into DCs and impaired their antigen-presenting ability. This resulted from a block of monocytes from entering the G1 phase of the cell cycle with a progressive number of cells accumulating in the G0 phase. Cyclin D2 was downregulated. However, differently from what was observed in T-cells stimulated in the presence of MSCs, the expression of p27 was found decreased, suggesting the involvement of similar but not identical pathways. CONCLUSIONS: We conclude that MSCs impair monocyte differentiation and function by interfering with the cell cycle. These findings imply that MSC-induced immunosuppression might be a side product of a more general antiproliferative effect. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17220794&query_hl=1 ER - TY - JFULL T1 - Substrate conformation modulates aggrecanase (ADAMTS-4) affinity and sequence specificity. Suggestion of a common topological specificity for functionally diverse proteases. A1 - Lauer-Fields, JL A1 - Minond, D A1 - Sritharan, T A1 - Kashiwagi, M A1 - Nagase, H A1 - Fields, GB J1 - J Biol Chem Y1 - 2007/01/05/ VL - 282 SN - 0021-9258 SP - 142 EP - 150 N2 - Protease-substrate interactions are governed by a variety of structural features. Although the substrate sequence specificities of numerous proteases have been established, "topological specificities," whereby proteases may be classified based on recognition of distinct three-dimensional structural motifs, have not. The aggrecanase members of the ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family cleave a variety of proteins but do not seem to possess distinct sequence specificities. In the present study, the topological substrate specificity of ADAMTS-4 (aggrecanase-1) was examined using triple-helical or single-stranded poly(Pro) II helical peptides. Substrate topology modulated the affinity and sequence specificity of ADAMTS-4 with K(m) values indicating a preference for triple-helical structure. In turn, non-catalytic ADAMTS-4 domains were critical for hydrolysis of triple-helical and poly(Pro) II helical substrates. Comparison of ADAMTS-4 with MMP-1 (collagenase 1), MMP-13 (collagenase 3), trypsin, and thermolysin using triple-helical peptide (THP) and single-stranded peptide (SSP) substrates demonstrated that all five proteases possessed efficient "triple-helical peptidase" activity and fell into one of two categories: (k(cat)/K(m))(SSP) > (k(cat)/K(m))(THP) (thermolysin, trypsin, and MMP-13) or (k(cat)/K(m))(THP) > or = (k(cat)/K(m))(SSP) and (K(m))(SSP) > (K(m))(THP) (MMP-1 and ADAMTS-4). Overall these results suggest that topological specificity may be a guiding principle for protease behavior and can be utilized to design specific substrates and inhibitors. The triple-helical and single-stranded poly(Pro) II helical peptides represent the first synthetic substrates successfully designed for aggrecanases. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17095512&query_hl=1 ER - TY - JFULL T1 - Collagen-induced arthritis in mice: a major role for tumor necrosis factor-alpha. A1 - Williams, RO J1 - Methods Mol Biol Y1 - 2007/// VL - 361 SN - 1064-3745 SP - 265 EP - 284 N2 - Collagen-induced arthritis is the most widely used animal model for the evaluation of novel therapeutic strategies for rheumatoid arthritis. The disease is induced by immunization of genetically susceptible strains of mice or rats with type II collagen in adjuvant. Susceptibility to collagen-induced arthritis is associated with major histocompatibility complex (MHC) class II genes, although non-MHC genes also play a role. Both B- and T-lymphocytes are important in the pathogenesis of collagen-induced arthritis, with the peak of the T-cell response occurring around the time of disease onset. Histopathological assessment of the joints of animals with collagen-induced arthritis reveal a proliferative synovitis with infiltration of polymorphonuclear and mononuclear cells, the formation of an erosive pannus, cartilage degradation, and fibrosis. As in human rheumatoid arthritis, a number of both pro- and anti-inflammatory cytokines are expressed in the joints of mice with collagen-induced arthritis, including tumor necrosis factor-alpha (TNFalpha) and interleukin (IL)-1beta, IL-6, IL-1Ra, IL-10, and transforming growth factor beta. The use transgenic and knockout strains of mice, as well as biological inhibitors, have revealed important pathological roles for multiple cytokines. Of these, TNFalpha emerged as a valid therapeutic target for rheumatoid arthritis and this led to the setting up of clinical trials of anti-TNFalpha antibody therapy. Three anti-TNFalpha biologics(infliximab, etanercept, and adalimumab) are now approved for use and TNFalpha blockade therefore represents an important advance in our ability to treat rheumatoid arthritis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17172717&query_hl=1 ER - TY - JFULL T1 - Apolipoprotein A-I breakdown is induced by thrombolysis in coronary patients. A1 - Eberini, I A1 - Gianazza, E A1 - Breghi, L A1 - Klugmann, S A1 - Calabresi, L A1 - Gomaraschi, M A1 - Mombelli, G A1 - Brusoni, B A1 - Wait, R A1 - Sirtori, CR J1 - Ann Med Y1 - 2007/// VL - 39 SN - 0785-3890 SP - 306 EP - 311 N2 - BACKGROUND: The outcome of percutaneous coronary intervention (PCI) is apparently worse in patients receiving a prior thrombolytic therapy ('facilitated PCI'). Recombinant tissue-type plasminogen activator (rt-PA) can degrade circulating high-density lipoproteins (HDL) bound apolipoprotein A-I (apoA-I), thus possibly reducing the vascular protective activity. There have never been reports of the detection of apolipoprotein breakdown products in the circulation. AIM: We studied the potential interactions between the protein components of HDL and tenecteplase, infused as thrombolytic therapy. METHODS: Sera from a total of 40 patients with acute myocardial infarction (AMI), unstable angina (UA), and dilative cardiomyopathy (controls) were investigated. AMI patients underwent either immediate PCI or were treated with tenecteplase thrombolysis. RESULTS: Products of extensive proteolysis of apoA-I were found in many acute coronary patients treated with tenecteplase, and in some AMI patients before starting the treatment (time 0). These were not detected in controls, UA patients as well as AMI patients undergoing immediate PCI. Small pre-beta-HDLs were selectively degraded. CONCLUSION: Significant apoA-I degradation occurs in AMI patients after thrombolytic treatment. This finding may provide a potential mechanism for the apparent reduction of benefit of facilitated versus nonfacilitated PCI. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17558602&query_hl=1 ER - TY - JFULL T1 - The pitfalls in the development of biologic therapy. A1 - Maini, RN A1 - Feldmann, M J1 - Nat Clin Pract Rheumatol Y1 - 2007/01// VL - 3 SN - 1745-8382 SP - 1 EP - 1 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17202998&query_hl=1 ER - TY - JFULL T1 - Cell therapy for autoimmune diseases. A1 - Dazzi, F A1 - van Laar, JM A1 - Cope, A A1 - Tyndall, A J1 - Arthritis Res Ther Y1 - 2007/// VL - 9 SN - 1478-6362 SP - 206 EP - 206 N2 - Cell therapy, pioneered for the treatment of malignancies in the form of bone marrow transplantation, has subsequently been tested and successfully employed in autoimmune diseases. Autologous haemopoietic stem cell transplantation (HSCT) has become a curative option for conditions with very poor prognosis such as severe forms of scleroderma, multiple sclerosis, and lupus, in which targeted therapies have little or no effect. The refinement of the conditioning regimens has virtually eliminated transplant-related mortality, thus making HSCT a relatively safe choice. Although HSCT remains a nonspecific approach, the knowledge gained in this field has led to the identification of new avenues. In fact, it has become evident that the therapeutic efficacy of HSCT cannot merely be the consequence of a high-dose immuno-suppression, but rather the result of a resetting of the abnormal immune regulation underlying autoimmune conditions. The identification of professional and nonprofessional immunosuppressive cells and their biological properties is generating a huge interest for their clinical exploitation. Regulatory T cells, found abnormal in several autoimmune diseases, have been proposed as central to achieve long-term remissions. Mesenchymal stem cells of bone marrow origin have more recently been shown not only to be able to differentiate into multiple tissues, but also to exert a potent antiproliferative effect that results in the inhibition of immune responses and prolonged survival of haemopoietic stem cells. All of these potential resources clearly need to be investigated at the preclinical level but support a great deal of enthusiasm for cell therapy of autoimmune diseases. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17367542&query_hl=1 ER - TY - JFULL T1 - HIF-2alpha is Essential for Hypoxic Induction of the Human Articular Chondrocyte Phenotype A1 - Lafont, J A1 - Talma, S A1 - Murphy, CL J1 - Arthritis and Rheumatism Y1 - 2007/// ER - TY - JFULL T1 - Crystal-ball gazing - the future of immunological research viewed from the cutting edge A1 - Brent, L A1 - Cohen, IR A1 - Doherty, PC A1 - Feldmann, M A1 - Matzinger, P A1 - Holgate, ST A1 - Lachmann, P A1 - Mitchison, NA A1 - Nossal, G A1 - Rose, NR A1 - Zinkernagel, R A1 - Ghost Lab J1 - CLIN EXP IMMUNOL Y1 - 2007/01// VL - 147 SN - 0009-9104 SP - 1 EP - 10 ER - TY - JFULL T1 - Molecular profile of peripheral blood mononuclear cells from patients with rheumatoid arthritis. A1 - Edwards, CJ A1 - Feldman, JL A1 - Beech, J A1 - Shields, KM A1 - Stover, JA A1 - Trepicchio, WL A1 - Larsen, G A1 - Foxwell, BM A1 - Brennan, FM A1 - Feldmann, M A1 - Pittman, DD J1 - Mol Med Y1 - 2007/01// VL - 13 SN - 1076-1551 SP - 40 EP - 58 N2 - Rheumatoid arthritis (RA) is a chronic inflammatory arthritis. Currently, diagnosis of RA may take several weeks, and factors used to predict a poor prognosis are not always reliable. Gene expression in RA may consist of a unique signature. Gene expression analysis has been applied to synovial tissue to define molecularly distinct forms of RA; however, expression analysis of tissue taken from a synovial joint is invasive and clinically impractical. Recent studies have demonstrated that unique gene expression changes can be identified in peripheral blood mononuclear cells (PBMCs) from patients with cancer, multiple sclerosis, and lupus. To identify RA disease-related genes, we performed a global gene expression analysis. RNA from PBMCs of 9 RA patients and 13 normal volunteers was analyzed on an oligonucleotide array. Compared with normal PBMCs, 330 transcripts were differentially expressed in RA. The differentially regulated genes belong to diverse functional classes and include genes involved in calcium binding, chaperones, cytokines, transcription, translation, signal transduction, extracellular matrix, integral to plasma membrane, integral to intracellular membrane, mitochondrial, ribosomal, structural, enzymes, and proteases. A k-nearest neighbor analysis identified 29 transcripts that were preferentially expressed in RA. Ten genes with increased expression in RA PBMCs compared with controls mapped to a RA susceptibility locus, 6p21.3. These results suggest that analysis of RA PBMCs at the molecular level may provide a set of candidate genes that could yield an easily accessible gene signature to aid in early diagnosis and treatment. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17515956&query_hl=1 ER - TY - JFULL T1 - Accuracy and reproducibility of protein-DNA microarray technology. A1 - Field, S A1 - Udalova, I A1 - Ragoussis, J J1 - Adv Biochem Eng Biotechnol Y1 - 2007/// VL - 104 SN - 0724-6145 SP - 87 EP - 110 N2 - Microarray-based methods for understanding protein-DNA interactions have been developed in the last 6 years due to the need to introduce high-throughput technologies in this field. Protein-DNA microarrays utilise chips upon which a large number of DNA sequences may be printed or synthesised. Any DNA-binding protein may then be interrogated by applying either purified sample or cellular/nuclear extracts, subject to availability of a suitable detection system. Protein is simply added to the microarray slide surface, which is then washed and subjected to at least one further incubation with a labelled molecule which binds specifically to the protein of interest. The signal obtained is proportional to the level of DNA-binding protein bound to each DNA feature, enabling relative affinities to be calculated. Key factors for reproducible and accurate quantification of protein binding are: microarray surface chemistry; length of oligonucleotides; position of the binding site sequence; quality of the protein and antibodies; and hybridisation conditions. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17290820&query_hl=1 ER - TY - JFULL T1 - Resuscitation of thermal injuries in the United Kingdom and Ireland. A1 - Baker, RH A1 - Akhavani, MA A1 - Jallali, N J1 - J Plast Reconstr Aesthet Surg Y1 - 2007/// VL - 60 SN - 1748-6815 SP - 682 EP - 685 N2 - The purpose of this study was to examine the consistency of burns resuscitation practice throughout UK and Ireland. Twenty-six Burns Units were identified via the National Burn Bed Bureau and surveyed via a postal questionnaire. Twenty-three units returned a completed questionnaire, covering all of the units treating children and 17 out of 20 units that treat adults. Nearly all of the Burns Units commence fluid resuscitation at 10% total body surface area of burn in children and 15% total body surface area of burn in adults. The estimated resuscitation volume is calculated using the Parkland or the Muir and Barclay formula in 76% and 11% of units, respectively. The most commonly used resuscitation fluid is Hartmann's solution. No unit uses blood as a first line fluid. Resuscitation is discontinued after 24h in 35% of units and after 36 h in 30% of units. Approximately half of the units do not routinely change the type of intravenous fluid administered after the initial period of resuscitation. This survey illustrates that resuscitation of thermally injured patients in UK and Ireland Burns Units is fairly consistent with a shift towards crystalloid resuscitation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17485059&query_hl=1 ER - TY - JFULL T1 - Localization of a long-range cis-regulatory element of IL13 by allelic transcript ratio mapping. A1 - Forton, JT A1 - Udalova, IA A1 - Campino, S A1 - Rockett, KA A1 - Hull, J A1 - Kwiatkowski, DP J1 - Genome Res Y1 - 2007/01// VL - 17 SN - 1088-9051 SP - 82 EP - 87 N2 - It appears that, for many genes, the two alleles possessed by an individual may produce different amounts of transcript. When such allelic differences in transcription are observed for some individuals but not others, a plausible explanation is genetic variation in the cis-acting elements that regulate the gene in question. Here we describe a novel analytical approach that uses such observations, combined with genotyping data from the HapMap project, to define the genomic location of cis-acting regulatory elements. When applied to the human 5q31 chromosomal region, where complex regulatory mechanisms are known to exist, we demonstrate the sensitivity of this approach by locating a highly significant cis-regulatory element operating on IL13 at long range from a position 250 kb upstream from the gene (P = 2 x 10(-6)). As this method is unaffected by other sources of variation, such as environmental and trans-acting genetic factors, it provides a tractable approach for dissecting the complexities of genetic variation in gene regulation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17135570&query_hl=1 ER - TY - JFULL T1 - The 'Hamburger' technique for harvesting cartilage grafts in nipple reconstruction. A1 - Norton, S A1 - Akhavani, MA A1 - Kang, N J1 - J Plast Reconstr Aesthet Surg Y1 - 2007/// VL - 60 SN - 1748-6815 SP - 957 EP - 959 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17616370&query_hl=1 ER - TY - JFULL T1 - The roles of substrate thermal stability and P-2 and P-1 ' subsite identity on matrix metalloproteinase triple-helical peptidase activity and collagen specificity A1 - Minond, D A1 - Lauer-Fields, JL A1 - Cudic, M A1 - Overall, CM A1 - Pei, DQ A1 - Brew, K A1 - Visse, R A1 - Nagase, H A1 - Fields, GB J1 - J BIOL CHEM Y1 - 2006/12/15/ VL - 281 SN - 0021-9258 SP - 38302 EP - 38313 N2 - The hydrolysis of collagen (collagenolysis) is one of the committed steps in extracellular matrix turnover. Within the matrix metalloproteinase (MMP) family distinct preferences for collagen types are seen. The substrate determinants that may guide these specificities are unknown. In this study, we have utilized 12 triple-helical substrates in combination with 10 MMPs to better define the contributions of substrate sequence and thermal stability toward triple helicase activity and collagen specificity. In general, MMP-13 was found to be distinct from MMP-8 and MT1-MMP(Delta 279-523), in that enhanced substrate thermal stability has only a modest effect on activity, regardless of sequence. This result correlates to the unique collagen specificity of MMP-13 compared with MMP-8 and MT1-MMP, in that MMP-13 hydrolyzes type II collagen efficiently, whereas MMP-8 and MT1-MMP are similar in their preference for type I collagen. In turn, MMP-1 was the least efficient of the collagenolytic MMPs at processing increasingly thermal stable triple helices and thus favors type III collagen, which has a relatively flexible cleavage site. Gelatinases (MMP-2 and MMP-9(Delta 444-707)) appear incapable of processing more stable helices and are thus mechanistically distinct from collagenolytic MMPs. The collagen specificity of MMPs appears to be based on a combination of substrate sequence and thermal stability. Analysis of the hydrolysis of triple-helical peptides by an MMP mutant indicated that Tyr(210) functions in triple helix binding and hydrolysis, but not in processing triple helices of increasing thermal stabilities. Further exploration of MMP active sites and exosites, in combination with substrate conformation, may prove valuable for additional dissection of collagenolysis and yield information useful in the design of more selective MMP inhibitors. ER - TY - JFULL T1 - Cell surface collagenolysis requires homodimerization of the membrane-bound collagenase MT1-MMP. A1 - Itoh, Y A1 - Ito, N A1 - Nagase, H A1 - Evans, RD A1 - Bird, SA A1 - Seiki, M J1 - Mol Biol Cell Y1 - 2006/12// VL - 17 SN - 1059-1524 SP - 5390 EP - 5399 N2 - Pericellular degradation of interstitial collagens is a crucial event for cells to migrate through the dense connective tissue matrices, where collagens exist as insoluble fibers. A key proteinase that participates in this process is considered to be membrane-type 1 matrix metalloproteinase (MT1-MMP or MMP-14), but little is known about the mechanism by which it cleaves the insoluble collagen. Here we report that homodimerization of MT1-MMP through its hemopexin (Hpx) domain is essential for cleaving type I collagen fibers at the cell surface. When dimerization was blocked by coexpressing either a membrane-bound or a soluble form of the Hpx domain, cell surface collagenolytic activity was inhibited in a dose-dependent manner. When MMP-13, a soluble collagenase active as a monomer in solution, was expressed as a membrane-anchored form on the cell surface, homodimerization was also required to cleave collagen. Our results introduce a new concept in that pericellular collagenolysis is regulated by correct molecular assembly of the membrane-anchored collagenase, thereby governing the directionality of the cell to migrate in tissue. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17050733&query_hl=1 ER - TY - JFULL T1 - Dual-specificity phosphatase 1: a critical regulator of innate immune responses. A1 - Abraham, SM A1 - Clark, AR J1 - Biochem Soc Trans Y1 - 2006/12// VL - 34 SN - 0300-5127 SP - 1018 EP - 1023 N2 - Innate immune responses are critically dependent on MAPK (mitogen-activated protein kinase) signalling pathways, in particular JNK (c-Jun N-terminal kinase) and p38 MAPK. Both of these kinases are negatively regulated via their dephosphorylation by DUSP1 (dual--specificity phosphatase 1). Several pro- and anti-inflammatory stimuli converge to regulate the DUSP1 gene and to modulate the time course of its expression. In turn, the pattern of expression of DUSP1 dictates the kinetics of activation of JNK and p38 MAPK, and this influences the expression of several mediators of innate immunity. DUSP1 is therefore a central regulator of innate immunity, and its expression can profoundly affect the outcome of inflammatory challenges. We discuss possible implications for immune-mediated inflammatory diseases and their treatment. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17073741&query_hl=1 ER - TY - JFULL T1 - Topographical variation in glycosaminoglycan content in human articular cartilage A1 - Rogers, BA A1 - Murphy, CL A1 - Cannon, SR A1 - Briggs, TWR J1 - J BONE JOINT SURG BR Y1 - 2006/12// VL - 88B SN - 0301-620X SP - 1670 EP - 1674 N2 - The weight-bearing status of articular cartilage has been shown to affect its biochemical composition. We have investigated the topographical variation of sulphated glycosaminoglycan (GAG) relative to the DNA content of the chondrocyte in human distal femoral articular cartilage.Paired specimens of distal femoral articular cartilage, from weight-bearing and non-weight-bearing regions, were obtained from 13 patients undergoing above-knee amputation. After papain enzyme digestion, spectrophotometric GAG and fluorometric DNA assays assessed the biochemical composition of the samples. The results were analysed using a paired t-test.Although there were no significant differences in cell density between the regions, the weight-bearing areas showed a significantly higher concentration of GAG relative to DNA when compared with non-weight-bearing areas (p = 0.02).We conclude that chondrocytes are sensitive to their mechanical environment, and that local loading conditions influence the metabolism of the cells and hence the biochemical structure of the tissue. ER - TY - JFULL T1 - The therapeutic potential of positive and negative immune cell co-stimulation during inflammation. A1 - Gwyer, E A1 - Snelgrove, R A1 - Hussell, T J1 - Biochem Soc Trans Y1 - 2006/12// VL - 34 SN - 0300-5127 SP - 1032 EP - 1036 N2 - Inflammatory cascades are initiated in response to alarm signals that may result from infection, malignant transformation or trauma. Immunity, however, must be controlled; otherwise damage may occur to otherwise healthy tissue within the same microenvironment. Similarly, peripheral tolerance mechanisms must ensure that autoreactive thymic or bone marrow emigrants do not respond upon encounter with the autoantigen. Organized lymphoid structures such as lymph nodes, spleen and Peyer's patches appear to regulate inflammation successfully, displaying controlled expansion and contraction. However, when immune cells flood into effector sites, the organization of T- and B-lymphocytes is lacking. What controls inflammatory cascades in lymph nodes but rarely in effector sites is not clear. We believe the difference lies in the Toll-like receptor ligand load, which is high in effector sites and drives uncontrolled inflammation. Similarly, we believe that initiation of autoimmune inflammation is initiated by the liberation of inflammatory signals due to infection or trauma. In this review, we highlight some of the molecules responsible for maintaining an activated T-cell phenotype, strategies to interrupt these therapeutically and the impact of ligating inhibitory receptors on antigen-presenting cells. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17073744&query_hl=1 ER - TY - JFULL T1 - Development of stable organic solvent nanofiltration membranes for membrane enhanced dynamic kinetic resolution A1 - Roengpithya, C A1 - Patterson, DA A1 - Taylor, PC A1 - Livingston, AG J1 - DESALINATION Y1 - 2006/11/20/ VL - 199 SN - 0011-9164 SP - 195 EP - 197 ER - TY - JFULL T1 - Pharmacological blockade of CCR1 ameliorates murine arthritis and alters cytokine networks in vivo A1 - Amat, M A1 - Benjamim, CF A1 - Williams, LM A1 - Prats, N A1 - Terricabras, E A1 - Beleta, J A1 - Kunkel, SL A1 - Godessart, N J1 - BRIT J PHARMACOL Y1 - 2006/11// VL - 149 SN - 0007-1188 SP - 666 EP - 675 N2 - Background and purpose: The chemokine receptor CCR1 is a potential target for the treatment of rheumatoid arthritis. To explore the impact of CCR1 blockade in experimental arthritis and the underlying mechanisms, we used J-113863, a non-peptide antagonist of the mouse receptor.Experimental approach: Compound J-113863 was tested in collagen-induced arthritis (CIA) and three models of acute inflammation; Staphylococcus enterotoxin B (SEB)-induced interleukin-2 (IL-2), delayed-type hypersensitivity (DTH) response, and lipopolysaccharide (LPS)-induced tumour necrosis factor alpha (TNF alpha) production. In the LPS model, CCR1 knockout, adrenalectomised, or IL-10-depleted mice were also used. Production of TNF alpha by mouse macrophages and human synovial membrane samples in vitro were also studied.Key results: Treatment of arthritic mice with J-113863 improved paw inflammation and joint damage, and dramatically decreased cell infiltration into joints. The compound did not inhibit IL-2 or DTH, but reduced plasma TNF alpha levels in LPS-treated mice. Surprisingly, CCR1 knockout mice produced more TNF alpha than controls in response to LPS, and J-113863 decreased TNFa also in CCR1 null mice, indicating that its effect was unrelated to CCR1. Adrenalectomy or neutralisation of IL-10 did not prevent inhibition of TNF alpha production by J-113863. The compound did not inhibit mouse TNF alpha in vitro, but did induce a trend towards increased TNF alpha release in cells from synovial membranes of rheumatoid arthritis patients.Conclusions and implications: CCR1 blockade improves the development of CIA, probably via inhibition of inflammatory cell recruitment. However, results from both CCR1-deficient mice and human synovial membranes suggest that, in some experimental settings, blocking CCR1 could enhance TNF production. ER - TY - JFULL T1 - Remission of collagen-induced arthritis is associated with high levels of transforming growth factor-beta expression in the joint. A1 - Marinova-Mutafchieva, L A1 - Gabay, C A1 - Funa, K A1 - Williams, RO J1 - Clin Exp Immunol Y1 - 2006/11// VL - 146 SN - 0009-9104 SP - 287 EP - 293 N2 - Immunization of genetically susceptible strains of mice with heterologous type II collagen leads to the induction of a self-limiting polyarthritis that begins to subside around 10 days after onset of clinical disease. The aims of this study were to compare pro- and anti-inflammatory cytokine expression in the joints during the course of arthritis in order to identify cytokines involved in spontaneous remission of arthritis. DBA/1 mice were immunized with type II collagen and an immunohistochemical analysis of expression of proinflammatory cytokines [tumour necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-6] and anti-inflammatory cytokines [IL-10, IL-1ra, transforming growth factor (TGF)-beta1, TGF-beta2 and TGF-beta3] in joints was carried out over the course of the disease. Both pro- and anti-inflammatory cytokines were found to be expressed in early arthritis. However, around 10 days after onset of arthritis, the level of expression of proinflammatory cytokines declined while the level of expression of anti-inflammatory cytokines, particularly TGF-beta1 and TGF-beta2, increased. Surprisingly, TNF-alpha continued to be expressed at low levels during the period of disease remission (30 days after onset). Blockade of TNF-alpha during the period of disease remission had no effect on TGF-beta expression. This study confirms that the level of inflammation in arthritis correlates strongly with the balance of pro- and anti-inflammatory cytokine expression in the joints. Of the anti-inflammatory cytokines studied, TGF-beta1 and TGF-beta2 predominate during the time of disease remission, suggesting that these cytokines are involved in regulating disease activity. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17034581&query_hl=1 ER - TY - JFULL T1 - A critical role for ICOS co-stimulation in immune containment of pulmonary influenza virus infection. A1 - Humphreys, IR A1 - Edwards, L A1 - Snelgrove, RJ A1 - Rae, AJ A1 - Coyle, AJ A1 - Hussell, T J1 - Eur J Immunol Y1 - 2006/11// VL - 36 SN - 0014-2980 SP - 2928 EP - 2938 N2 - Lung pathology observed during influenza infection is due to direct damage resulting from viral replication and bystander damage caused by overly exuberant antiviral immune mechanisms. In the absence of universally effective vaccines and antiviral therapies, knowledge of the cellular components required for immune containment of influenza is essential. ICOS is a late co-stimulatory molecule expressed by T cells 12-24 h after activation. We show for the first time that inhibition of ICOS with a monoclonal antibody reduces pulmonary T cell inflammation and associated cytokine expression. Surprisingly however, this reduction in T cells was not accompanied by an alleviation of weight loss and illness. Furthermore, lung viral titres were elevated following anti-ICOS treatment, suggesting that the beneficial outcome of reducing T cell pathology was masked by enhanced virus-induced damage and innate inflammation. This study demonstrates the delicate balance that exists between pathogen burden and pulmonary T cell inflammation during influenza infection and highlights the critical role of ICOS in this response. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17039567&query_hl=1 ER - TY - JFULL T1 - The RNA binding protein Zfp36l1 is required for normal vascularisation and post-transcriptionally regulates VEGF expressiono A1 - Bell, SE A1 - Sanchez, MJ A1 - Spasic-Boskovic, O A1 - Santalucia, T A1 - Gambardella, L A1 - Burton, GJ A1 - Murphy, JJ A1 - Norton, JD A1 - Clark, AR A1 - Turner, M J1 - DEV DYNAM Y1 - 2006/11// VL - 235 SN - 1058-8388 SP - 3144 EP - 3155 N2 - The Zfp36l1 gene encodes a zinc finger-containing mRNA binding protein implicated in the posttranscriptional control of gene expression. Mouse embryos homozygous for a targeted mutation in the Zfp36l1 locus died mid-gestation and exhibited extraembryonic and intraembryonic vascular abnormalities and heart defects. In the developing placenta, there was a failure of the extraembryonic mesoderm to invaginate the trophoblast layer. The phenotype was associated with an elevated expression of vascular endothelial growth factor (VEGF)-A in the embryos and in embryonic fibroblasts cultured under conditions of both normoxia and hypoxia. VEGF-A overproduction by embryonic fibroblasts was not a consequence of changes in Vegf-a mRNA stability; instead, we observed enhanced association with polyribosomes, suggesting Zfp36l1 influences translational regulation. These data implicate Zfp36l1 as a negative regulator of Vegf-a gene activity during development. ER - TY - JFULL T1 - Chain register shift in type I collagen alters MMP1 cleavage A1 - Kuznetsova, NV A1 - Makareeva, E A1 - Cabral, WA A1 - Visse, R A1 - Nagase, H A1 - Marini, JC A1 - Leikin, S J1 - MATRIX BIOL Y1 - 2006/11// VL - 25 SN - 0945-053X SP - S75 EP - S75 ER - TY - JFULL T1 - Direct activation of type I protein kinase a (PKA) by oxidants independently of cAMP is mediated by RI subunit interprotein disulphide bond formation A1 - Brennan, JP A1 - Bardswell, SC A1 - Burgoyne, JR A1 - Fuller, W A1 - Schroder, E A1 - Wait, R A1 - Begum, S A1 - Kentish, JC A1 - Eaton, P J1 - CIRCULATION Y1 - 2006/10/31/ VL - 114 SN - 0009-7322 SP - 1 EP - 2 ER - TY - JFULL T1 - In the absence of reactive oxygen species, T cells default to a Th1 phenotype and mediate protection against pulmonary Cryptococcus neoformans infection. A1 - Snelgrove, RJ A1 - Edwards, L A1 - Williams, AE A1 - Rae, AJ A1 - Hussell, T J1 - J Immunol Y1 - 2006/10/15/ VL - 177 SN - 0022-1767 SP - 5509 EP - 5516 N2 - In recent years, the prevalence of invasive fungal infections has increased, attributed mostly to the rising population of immunocompromised individuals. Cryptococcus neoformans has been one of the most devastating, with an estimated 6-8% of AIDS-infected patients succumbing to Cryptococcus-associated meningitis. Reactive oxygen species (ROS) are potent antimicrobial agents but also play a significant role in regulating immune cell phenotype, but cause immunopathology when produced in excess. We now show that mice lacking phagocyte NADPH oxidase have heightened macrophage and Th1 responses and improved pathogen containment within pulmonary granulomatous lesions. Consequently, dissemination of this fungus to the brain is diminished, an effect that is independent of IL-12. Similar results are described using the metalloporphyrin antioxidant manganese(III) tetrakis(N-ethyl pyridinium-2-yl)porphyrin, which also promoted a protective Th1 response and reduced dissemination to the brain. These findings are in sharp contrast to the protective potential of ROS against other fungal pathogens, and highlight the pivotal role that ROS can fulfill in shaping the profile of the host's immune response. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17015737&query_hl=1 ER - TY - JFULL T1 - Adenoviral gene transfer into osteoarthritis synovial cells using the endogenous inhibitor I kappa B alpha reveals that most, but not all, inflammatory and destructive mediators are NF kappa B dependent A1 - Amos, N A1 - Lauder, S A1 - Evans, A A1 - Feldmann, M A1 - Bondeson, J J1 - RHEUMATOLOGY Y1 - 2006/10// VL - 45 SN - 1462-0324 SP - 1201 EP - 1209 N2 - Objectives. Despite recent major advances in the understanding of the pathogenesis of rheumatoid arthritis, with tumour necrosis factor-alpha (TNF alpha) established as a major therapeutic target, comparatively little is known about the mediators involved in the destructive and inflammatory pathways in osteoarthritis (OA). Recently, it has become appreciated that an inflammatory synovitis contributes not only to the signs and symptoms of osteoarthritis, but also to disease progression. Here, we use high-efficiency adenoviral gene transfer to investigate the role of the transcription factor nuclear factor-kappa B (NF kappa B) in regulating inflammatory and destructive mediators in the late stage OA synovium.Methods. Infection with reporter adenoviruses transferring the beta-galactosidase or green fluorescent protein genes verified that OA synovial cells could be infected (> 95%). Adenovirus transferring the inhibitory subunit I kappa B alpha inhibited NF kappa B. The production of a whole array of pro-and anti-inflammatory cytokines and mediators, and several matrix metalloproteinases and their main inhibitor, was measured by enzyme-linked immunosorbent assay.Results. The spontaneous production of macrophage-produced pro-inflammatory cytokines varied: TNF alpha was modestly inhibited by I kappa B alpha overexpression, but interleukin (IL)-1 was unaffected. Both IL-6 and IL-8 were potently inhibited, as were granulocyte-macrophage colony stimulating factor and oncostatin M. Anti-inflammatory mediators like IL-10, the IL-1 receptor antagonist and the p55 soluble TNF receptor were unaffected. Matrix metalloproteinases 1, 3, 9 and 13 were potently inhibited by I kappa B alpha overexpression, but not their main inhibitor tissue inhibitor of metalloproteinase-1.Conclusions. The OA synovium is a highly inflammatory environment, with spontaneous production of many cytokines and matrix metalloproteinases. Inhibition of NF kappa B may have a beneficial effect on the balance between pro-inflammatory cytokines and anti-inflammatory mediators, and between destructive metalloproteinases and their main inhibitor. ER - TY - JFULL T1 - Should we be using intraarticular tumor necrosis factor blockade in inflammatory monoarthritis? A1 - Fisher, BA A1 - Keat, A J1 - J Rheumatol Y1 - 2006/10// VL - 33 SN - 0315-162X SP - 1934 EP - 1935 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17014012&query_hl=1 ER - TY - JFULL T1 - MT1-MMP: a key regulator of cell migration in tissue. A1 - Itoh, Y J1 - IUBMB Life Y1 - 2006/10// VL - 58 SN - 1521-6543 SP - 589 EP - 596 N2 - Controlled cell migration is a fundamental and critical event in many physiological processes. However once control is lost, cell migration facilitates disease progression such as seen in cancer metastasis, atherosclerosis, and rheumatoid arthritis. One of the critical proteinases involved in cell migration is membrane-type 1 matrix metalloproteinase (MT1-MMP/MMP-14). MT1-MMP degrades extracellular matrix to make a path for cells to migrate, sheds cell surface molecules to give migratory signals, and activates ERK (extracellular signal-regulated protein kinase) enhancing cell migration. For MT1-MMP to promote cell migration, it needs to act in co-ordination with other cell migration machinery. Understanding such regulatory links may provide insights into the development of novel disease therapies. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17050376&query_hl=1 ER - TY - JFULL T1 - Parallel protein and transcript profiles of FSHD patient muscles correlate to the D4Z4 arrangement and reveal a common impairment of slow to fast fibre differentiation and a general deregulation of MyoD-dependent genes. A1 - Celegato, B A1 - Capitanio, D A1 - Pescatori, M A1 - Romualdi, C A1 - Pacchioni, B A1 - Cagnin, S A1 - Viganò, A A1 - Colantoni, L A1 - Begum, S A1 - Ricci, E A1 - Wait, R A1 - Lanfranchi, G A1 - Gelfi, C J1 - Proteomics Y1 - 2006/10// VL - 6 SN - 1615-9853 SP - 5303 EP - 5321 N2 - Here, we present the first study of a human neuromuscular disorder at transcriptional and proteomic level. Autosomal dominant facio-scapulo-humeral muscular dystrophy (FSHD) is caused by a deletion of an integral number of 3.3-kb KpnI repeats inside the telomeric region D4Z4 at the 4q35 locus. We combined a muscle-specific cDNA microarray platform with a proteomic investigation to analyse muscle biopsies of patients carrying a variable number of KpnI repeats. Unsupervised cluster analysis divides patients into three classes, according to their KpnI repeat number. Expression data reveal a transition from fast-glycolytic to slow-oxidative phenotype in FSHD muscle, which is accompanied by a deficit of proteins involved in response to oxidative stress. Besides, FSHD individuals show a disruption in the MyoD-dependent gene network suggesting a coregulation at transcriptional level during myogenesis. We also discuss the hypothesis that D4Z4 contraction may affect in trans the expression of a set of genes involved in myogenesis, as well as in the regeneration pathway of satellite cells in adult tissue. Muscular wasting could result from the inability of satellite cells to successfully differentiate into mature fibres and from the accumulation of structural damages caused by a reactive oxygen species (ROS) imbalance induced by an increased oxidative metabolism in fibres. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17013991&query_hl=1 ER - TY - JFULL T1 - Activation of NF-kappaB by the intracellular expression of NF-kappaB-inducing kinase acts as a powerful vaccine adjuvant. A1 - Andreakos, E A1 - Williams, RO A1 - Wales, J A1 - Foxwell, BM A1 - Feldmann, M J1 - Proc Natl Acad Sci U S A Y1 - 2006/09/26/ VL - 103 SN - 0027-8424 SP - 14459 EP - 14464 N2 - There is a pressing need for adjuvants that will enhance the effectiveness of genetic vaccines. This is particularly important in cancer and infectious disease such as HIV and malaria for which successful vaccines are desperately needed. Here, we describe an approach to enhance immunogenicity that involves the activation of NF-kappaB by the transgenic expression of an intracellular signaling molecule, NF-kappaB-inducing kinase (NIK). In vitro, NIK increases dendritic cell antigen presentation in allogeneic and antigen-specific T cell proliferation assays by potently activating NF-kappaB and consequently up-regulating the expression of cytokines (TNF-alpha, IL-6, IL-12, IL-15, and IL-18), chemokines [IL-8, RANTES (regulated on activation, normal T cell expressed and secreted), macrophage inflammatory protein-1alpha, monocyte chemoattractant protein-1, and monocyte chemoattractant protein-3], MHC antigen-presenting molecules (class I and II), and costimulatory molecules (CD80 and CD86). In vivo, NIK enhances immune responses against a vector-encoded antigen and shifts them toward a T helper 1 immune response with increased IgG2a levels, T cell proliferation, IFN-gamma production, and cytotoxic T lymphocyte responses more potently than complete Freund's adjuvant, a very efficacious T helper 1-inducing adjuvant. These findings define NIK, and possibly other inducers of NF-kappaB activation, as a potent adjuvant strategy that offers great potential for genetic vaccine development. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16971487&query_hl=1 ER - TY - JFULL T1 - Free innervated sole of foot transfer for contralateral lower limb salvage. A1 - Irwin, MS A1 - Jain, A A1 - Anand, P A1 - Nanchahal, J J1 - Plast Reconstr Surg Y1 - 2006/09/15/ VL - 118 SN - 1529-4242 SP - 93e EP - 97e L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16980840&query_hl=1 ER - TY - JFULL T1 - TIMP-3 inhibits the procollagen N-proteinase ADAMTS-2 A1 - Wang, WM A1 - Ge, GX A1 - Lim, NH A1 - Nagase, H A1 - Greenspan, DS J1 - BIOCHEM J Y1 - 2006/09/15/ VL - 398 SN - 0264-6021 SP - 515 EP - 519 N2 - ADAMTS-2 is an extracellular metalloproteinase responsible for cleaving the N-propeptides of procollagens I-III; an activity necessary for the formation of collagenous ECM (extracellular matrix). The four TIMPs (tissue inhibitors of metalloprotemases) regulate the activities of matrix metalloproteinases, which are involved in degrading ECM components. Here we delineate the abilities of the TIMPs to affect biosynthetic processing of procollagens. TIMP-1, -2 and -4 show no inhibitory activity towards ADAMTS-2, in addition none of the TIMPs showed inhibitory activity towards bone morphogenetic protein 1, which is responsible for cleaving procollagen C-propeptides. In contrast, TIMP-3 is demonstrated to inhibit ADAMTS-2 in vitro with apparent K-i values of 160 and 602 nM, in the presence of heparin or without respectively; and TIMP-3 is shown to inhibit procollagen processing by cells. ER - TY - JFULL T1 - Crystal structure of an active form of human MMP-1 A1 - Iyer, S A1 - Visse, R A1 - Nagase, H A1 - Acharya, KR J1 - J MOL BIOL Y1 - 2006/09/08/ VL - 362 SN - 0022-2836 SP - 78 EP - 88 N2 - The extracellular matrix is a dynamic environment that constantly undergoes remodelling and degradation during vital physiological processes such as angiogenesis, wound healing, and development. Unbalanced extracellular matrix breakdown is associated with many diseases such as arthritis, cancer and fibrosis. Interstitial collagen is degraded by matrix metalloproteinases with collagenolytic activity by MMP-1, MMP-8 and MMP-13, collectively known as the collagenases. Matrix metalloproteinase 1 (MMP-1) plays a pivotal role in degradation of interstitial collagen types I, II, and III. Here, we report the crystal structure of the active form of human MMP-1 at 2.67 angstrom resolution. This is the first MMP-1 structure that is free of inhibitor and a water molecule essential for peptide hydrolysis is observed coordinated with the active site zinc. Comparing this structure with the human proMMP-1 shows significant structural differences, mainly in the relative orientation of the hemopexin domain, between the pro form and active form of the human enzyme. (c) 2006 Elsevier Ltd. All rights reserved. ER - TY - JFULL T1 - TLR4-NOX4-AP-1 signaling mediates lipopolysaccharide-induced CXCR6 expression in human aortic smooth muscle cells. A1 - Patel, DN A1 - Bailey, SR A1 - Gresham, JK A1 - Schuchman, DB A1 - Shelhamer, JH A1 - Goldstein, BJ A1 - Foxwell, BM A1 - Stemerman, MB A1 - Maranchie, JK A1 - Valente, AJ A1 - Mummidi, S A1 - Chandrasekar, B J1 - Biochem Biophys Res Commun Y1 - 2006/09/08/ VL - 347 SN - 0006-291X SP - 1113 EP - 1120 N2 - CXCL16 is a transmembrane non-ELR CXC chemokine that signals via CXCR6 to induce aortic smooth muscle cell (ASMC) proliferation. While bacterial lipopolysaccharide (LPS) has been shown to stimulate CXCL16 expression in SMC, its effects on CXCR6 are not known. Here, we demonstrate that LPS upregulates CXCR6 mRNA, protein, and surface expression in human ASMC. Inhibition of TLR4 with neutralizing antibodies or specific siRNA interference blocked LPS-mediated CXCR6 expression. LPS stimulated both AP-1 (c-Fos, c-Jun) and NF-kappaB (p50 and p65) activation, but only inhibition of AP-1 attenuated LPS-induced CXCR6 expression. Using dominant negative expression vectors and siRNA interference, we demonstrate that LPS induces AP-1 activation via MyD88, TRAF6, ERK1/2, and JNK signaling pathways. Furthermore, the flavoprotein inhibitor diphenyleniodonium chloride significantly attenuated LPS-mediated AP-1-dependent CXCR6 expression, as did inhibition of NOX4 NADPH oxidase by siRNA. Finally, CXCR6 knockdown inhibited CXCL16-induced ASMC proliferation. These results demonstrate that LPS-TLR4-NOX4-AP-1 signaling can induce CXCR6 expression in ASMC, and suggest that the CXCL16-CXCR6 axis may be an important proinflammatory pathway in the pathogenesis of atherosclerosis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16870145&query_hl=1 ER - TY - JFULL T1 - Development of a novel 2D proteomics approach for the identification of proteins secreted by primary chondrocytes after stimulation by IL-1 and oncostatin M. A1 - Catterall, JB A1 - Rowan, AD A1 - Sarsfield, S A1 - Saklatvala, J A1 - Wait, R A1 - Cawston, TE J1 - Rheumatology (Oxford) Y1 - 2006/09// VL - 45 SN - 1462-0324 SP - 1101 EP - 1109 N2 - OBJECTIVES: To develop a proteomics approach to study changes in the secreted protein levels of primary human chondrocytes after stimulation by the pro-inflammatory cytokines interleukin-1 and oncostatin M. METHODS: Using both the primary human articular and bovine nasal chondrocyte-conditioned mediums, methods were investigated to enable the separation of proteins by two-dimensional (2D) gel electrophoresis. Differentially regulated proteins were identified using tandem electrospray mass spectrometery. RESULTS: We discovered that proteoglycans and glycosylaminoglycans (GAGs) secreted by chondrocytes significantly interfered with 2D gel focusing. Several different methods for GAG removal were attempted including enzymic digestion, cetyl pyridinium chloride precipitation and anion exchange in high salt. The anion exchange proved to be the most effective. Even from these initial gels, we were able to identify eight proteins produced by human chondrocytes: matrix metalloproteinase (MMP)-1, MMP-3, YKL40, cyclophilin A, beta2-microglobulin, transthyretin, S100A11, peroxidine 1 and cofilin. MMP-1, MMP-3, YKL40 and cyclophilin A were all identified as processed, smaller peptide fragments. CONCLUSIONS: We were able to develop a novel sample preparation protocol to allow the reproducible sample preparation of secreted proteins from human chondrocytes. From the initial data, we were able to show that at least some of the proteins produced were cleaved to smaller fragments as a result of proteolysis. Therefore, this technique provides valuable information about protein processing which gene-based arrays do not. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16567360&query_hl=1 ER - TY - JFULL T1 - Immunohistochemical expression of hypoxic inducible factor (HIF) and vascular endothelial growth factor (VEGF) in colorectal cancer patients on statin therapy A1 - De Four, K A1 - Paleolog, E A1 - Sandison, A A1 - Dawson, P A1 - Cohen, P J1 - MODERN PATHOL Y1 - 2006/09// VL - 19 SN - 0893-3952 SP - 57 EP - 57 ER - TY - JFULL T1 - Structural features of the reprolysin atrolysin C and tissue inhibitors of metalloproteinases (TIMPs) interaction A1 - Pinto, AFM A1 - Terra, RMS A1 - Guimaraes, JA A1 - Kashiwagi, M A1 - Nagase, H A1 - Serrano, SMT A1 - Fox, JW J1 - BIOCHEM BIOPH RES CO Y1 - 2006/09/01/ VL - 347 SN - 0006-291X SP - 641 EP - 648 N2 - Atrolysin C is a P-I snake venom metalloproteinase (SVMP) from Crotalus atrox venom, which efficiently degrades capillary basement membranes, extracellular matrix, and cell surface proteins to produce hemorrhage. The tissue inhibitors of metalloproteinases (TIMPs) are effective inhibitors of matrix metalloproteinases which share some structural similarity with the SVMPs. In this work, we evaluated the inhibitory profile of TIMP-1, TIMP-2, and the N-terminal domain of TIMP-3 (N-TIMP-3) on the proteolytic activity of atrolysin C and analyzed the structural requirements and molecular basis of inhibitor-enzyme interaction using molecular modeling. While TIMP-1 and TIMP-2 had no inhibitory activity upon atrolysin C, the N-terminal domain of TIMP-3 (N-TIMP-3) was a potent inhibitor with a K-i value of approximately 150 nM. The predicted docking structures of atrolysin C and TIMPs were submitted to molecular dynamics simulations and the complex atrolysin C/N-TIMP-3 was the only one that maintained the inhibitory conformation. This study is the first to shed light on the structural determinants required for the interaction between a SVMP and a TIMP, and suggests a structural basis for TIMP-3 inhibitory action and related proteins such as the ADAMs. (c) 2006 Elsevier Inc. All rights reserved. ER - TY - JFULL T1 - Double-blind randomized controlled clinical trial of the interleukin-6 receptor antagonist, tocilizumab, in European patients with rheumatoid arthritis who had an incomplete response to methotrexate. A1 - Maini, RN A1 - Taylor, PC A1 - Szechinski, J A1 - Pavelka, K A1 - Bröll, J A1 - Balint, G A1 - Emery, P A1 - Raemen, F A1 - Petersen, J A1 - Smolen, J A1 - Thomson, D A1 - Kishimoto, T A1 - CHARISMA Study Group J1 - Arthritis Rheum Y1 - 2006/09// VL - 54 SN - 0004-3591 SP - 2817 EP - 2829 N2 - OBJECTIVE: To establish the safety and efficacy of repeat infusions of tocilizumab (previously known as MRA), a humanized anti-interleukin-6 (IL-6) receptor antibody, alone and in combination with methotrexate (MTX), for the treatment of rheumatoid arthritis (RA). METHODS: The study group comprised 359 patients with active RA in whom the response to MTX was inadequate. During a stabilization period, these patients received their current dose of MTX for at least 4 weeks. Following stabilization, they were randomized to 1 of 7 treatment arms, as follows: tocilizumab at doses of 2 mg/kg, 4 mg/kg, or 8 mg/kg either as monotherapy or in combination with MTX, or MTX plus placebo. RESULTS: A 20% response (improvement) according to the American College of Rheumatology criteria (ACR20 response) was achieved by 61% and 63% of patients receiving 4 mg/kg and 8 mg/kg of tocilizumab as monotherapy, respectively, and by 63% and 74% of patients receiving those doses of tocilizumab plus MTX, respectively, compared with 41% of patients receiving placebo plus MTX. Statistically significant ACR50 and ACR70 responses were observed in patients receiving combination therapy with either 4 mg/kg or 8 mg/kg of tocilizumab plus MTX (P < 0.05). A dose-related reduction in the Disease Activity Score in 28 joints was observed from week 4 onward, in all patients except those receiving monotherapy with 2 mg/kg of tocilizumab. In the majority of patients who received 8 mg/kg of tocilizumab, the C-reactive protein level/erythrocyte sedimentation rate normalized, while placebo plus MTX had little effect on these laboratory parameters. Tocilizumab was mostly well tolerated, with a safety profile similar to that of other biologic and immunosuppressive therapies. Alanine transaminase and aspartate transaminase levels followed a sawtooth pattern (rising and falling between infusions). There were moderate but reversible increases in the nonfasting total cholesterol and triglyceride levels and reversible reductions in the high-density lipoprotein cholesterol and neutrophil levels. There were 2 cases of sepsis, both of which occurred in patients who were receiving combination therapy with 8 mg/kg of tocilizumab plus MTX. CONCLUSION: These results indicate that targeted blockade of IL-6 signaling is a highly efficacious and promising means of decreasing disease activity in RA. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16947782&query_hl=1 ER - TY - JFULL T1 - Infliximab inhibits radiographic progression regardless of disease activity at baseline and following treatment in patients with early active rheumatoid arthritis. A1 - Smolen, JS A1 - Han, C A1 - van der Heijde, D A1 - Emery, P A1 - Bathon, JM A1 - Keystone, E A1 - Maini, RN A1 - Kalden, JR A1 - Aletaha, D A1 - Baker, D A1 - Han, J A1 - Bala, M A1 - Clair, EW J1 - ARTHRITIS RHEUM Y1 - 2006/09// VL - 54 SN - 0004-3591 SP - S231 EP - S231 ER - TY - JFULL T1 - Harmful waste products as novel immune modulators for treating inflammatory arthritis? A1 - Cope, AP J1 - PLoS Med Y1 - 2006/09// VL - 3 SN - 1549-1676 SP - e385 EP - e385 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16968131&query_hl=1 ER - TY - JFULL T1 - Modulation of Cell-Fibronectin Matrix Interactions during Tissue Repair. A1 - Midwood, KS A1 - Mao, Y A1 - Hsia, HC A1 - Valenick, LV A1 - Schwarzbauer, JE J1 - J Invest Dermatol Y1 - 2006/09// VL - 126 Suppl SN - 0022-202X SP - 73 EP - 78 N2 - Environmental signals from the extracellular matrix (ECM) are transmitted by cell surface receptors that connect to the actin cytoskeleton and to multiple intracellular signaling pathways. To dissect how the ECM regulates cell functions, we are using a three-dimensional (3D) fibrin-fibronectin matrix, resembling the wound provisional matrix. Fibroblasts adhere to fibronectin in this matrix via concomitant engagement of alpha5beta1 integrin receptors and syndecan-4, a transmembrane proteoglycan. An adhesive phenotype is developed with actin stress fibers and activation of focal adhesion kinase (FAK) and Rho GTPase. Lack of syndecan-4 engagement, as occurs in the presence of the ECM protein tenascin-C, promotes a motile phenotype; FAK and Rho signaling are downregulated and filopodia are extended. Fibronectin matrices have distinct effects on two other receptors: alpha4beta1 and alphavbeta3 integrins. Although alpha4beta1 does not naturally support strong cell interactions with a fibrin-fibronectin matrix, binding is dramatically enhanced by proteolytic cleavage of fibronectin. Conversely, activity of alphavbeta3 is stimulated by multimeric fibronectin fibrils showing that the organization of fibronectin differentially affects integrin functions. Thus, deposition of additional ECM components, expression of co-receptors for ECM, cleavage of adhesive proteins, and the architecture of the ECM microenvironment are different mechanisms for modulating cell responses to fibronectin matrix.Journal of Investigative Dermatology Symposium Proceedings (2006) 11, 73-78. doi:10.1038/sj.jidsymp.5650005. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16983359&query_hl=1 ER - TY - JFULL T1 - The synovial fluid: a site for citrullinated autoantigen generation in RA. A1 - Kinloch, A A1 - Lundberg, K A1 - Begum, S A1 - Peirce, MJ A1 - Wait, R A1 - Venables, PJ J1 - ARTHRITIS RHEUM Y1 - 2006/09// VL - 54 SN - 0004-3591 SP - S566 EP - S567 ER - TY - JFULL T1 - Antiinflammatory effects of dexamethasone are partly dependent on induction of dual specificity phosphatase 1. A1 - Abraham, SM A1 - Lawrence, T A1 - Kleiman, A A1 - Warden, P A1 - Medghalchi, M A1 - Tuckermann, J A1 - Saklatvala, J A1 - Clark, AR J1 - J Exp Med Y1 - 2006/08/07/ VL - 203 SN - 0022-1007 SP - 1883 EP - 1889 N2 - Glucocorticoids (GCs), which are used in the treatment of immune-mediated inflammatory diseases, inhibit the expression of many inflammatory mediators. They can also induce the expression of dual specificity phosphatase 1 (DUSP1; otherwise known as mitogen-activated protein kinase [MAPK] phosphatase 1), which dephosphorylates and inactivates MAPKs. We investigated the role of DUSP1 in the antiinflammatory action of the GC dexamethasone (Dex). Dex-mediated inhibition of c-Jun N-terminal kinase and p38 MAPK was abrogated in DUSP1-/- mouse macrophages. Dex-mediated suppression of several proinflammatory genes (including tumor necrosis factor, cyclooxygenase 2, and interleukin 1alpha and 1beta) was impaired in DUSP1-/- mouse macrophages, whereas other proinflammatory genes were inhibited by Dex in a DUSP1-independent manner. In vivo antiinflammatory effects of Dex on zymosan-induced inflammation were impaired in DUSP1-/- mice. Therefore, the expression of DUSP1 is required for the inhibition of proinflammatory signaling pathways by Dex in mouse macrophages. Furthermore, DUSP1 contributes to the antiinflammatory effects of Dex in vitro and in vivo. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16880258&query_hl=1 ER - TY - JFULL T1 - Oxidant-induced activation of type I protein kinase A is mediated by RI subunit interprotein disulfide bond formation. A1 - Brennan, JP A1 - Bardswell, SC A1 - Burgoyne, JR A1 - Fuller, W A1 - Schröder, E A1 - Wait, R A1 - Begum, S A1 - Kentish, JC A1 - Eaton, P J1 - J Biol Chem Y1 - 2006/08/04/ VL - 281 SN - 0021-9258 SP - 21827 EP - 21836 N2 - Here we demonstrate that type I protein kinase A is redoxactive, forming an interprotein disulfide bond between its two regulatory RI subunits in response to cellular hydrogen peroxide. This oxidative disulfide formation causes a subcellular translocation and activation of the kinase, resulting in phosphorylation of established substrate proteins. The translocation is mediated at least in part by the oxidized form of the kinase having an enhanced affinity for alpha-myosin heavy chain, which serves as a protein kinase A (PKA) anchor protein and localizes the PKA to its myofilament substrates troponin I and myosin binding protein C. The functional consequence of these events in cardiac myocytes is that hydrogen peroxide increases contractility independently of beta-adrenergic stimulation and elevations of cAMP. The oxidant-induced phosphorylation of substrate proteins and increased contractility is blocked by the kinase inhibitor H89, indicating that these events involve PKA activation. In essence, type I PKA contains protein thiols that operate as redox sensors, and their oxidation by hydrogen peroxide directly activates the kinase. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16754666&query_hl=1 ER - TY - JFULL T1 - Cytokines for myocardial ischaemia: a echocardiographic study A1 - Raisakis, K A1 - Monaco, C A1 - Stamatelopoulos, K A1 - Desforges, L A1 - Liodakis, E A1 - Nihoyannopoulos, P J1 - EUR HEART J Y1 - 2006/08// VL - 27 SN - 0195-668X SP - 561 EP - 561 ER - TY - JFULL T1 - Survey on the perioperative use of TNFalpha inhibitors in rheumatoid hand surgery. A1 - Jain, A A1 - Harley, O A1 - Patel, RM A1 - Nanchahal, J J1 - J Hand Surg [Br] Y1 - 2006/08// VL - 31 SN - 0266-7681 SP - 463 EP - 464 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16716465&query_hl=1 ER - TY - JFULL T1 - TF ligation by factor VIIa induces pro-inflammatory cytokine production in atheroma smooth muscle cells A1 - Persson, L A1 - Navin, T A1 - Gregan, S A1 - Davies, AH A1 - Feldmann, M A1 - Monaco, C J1 - EUR HEART J Y1 - 2006/08// VL - 27 SN - 0195-668X SP - 455 EP - 455 ER - TY - JFULL T1 - Role of CCL5 (RANTES) in viral lung disease. A1 - Culley, FJ A1 - Pennycook, AM A1 - Tregoning, JS A1 - Dodd, JS A1 - Walzl, G A1 - Wells, TN A1 - Hussell, T A1 - Openshaw, PJ J1 - J Virol Y1 - 2006/08// VL - 80 SN - 0022-538X SP - 8151 EP - 8157 N2 - CCL5/RANTES is a key proinflammatory chemokine produced by virus-infected epithelial cells and present in respiratory secretions of asthmatics. To examine the role of CCL5 in viral lung disease, we measured its production during primary respiratory syncytial virus (RSV) infection and during secondary infection after sensitizing vaccination that induces Th2-mediated eosinophilia. A first peak of CCL5 mRNA and protein production was seen at 18 to 24 h of RSV infection, before significant lymphocyte recruitment occurred. Treatment in vivo with Met-RANTES (a competitive chemokine receptor blocker) throughout primary infection decreased CD4+ and CD8+ cell recruitment and increased viral replication. In RSV-infected, sensitized mice with eosinophilic disease, CCL5 production was further augmented; Met-RANTES treatment again reduced inflammatory cell recruitment and local cytokine production. A second wave of CCL5 production occurred on day 7, attributable to newly recruited T cells. Paradoxically, mice treated with Met-RANTES during primary infection demonstrated increased cellular infiltration during reinfection. We therefore show that RSV induces CCL5 production in the lung and this causes the recruitment of RSV-specific cells, including those making additional CCL5. If this action is blocked with Met-RANTES, inflammation decreases and viral clearance is delayed. However, the exact effects of chemokine modulation depend critically on time of administration, a factor that may potentially complicate the use of chemokine blockers in inflammatory diseases. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16873271&query_hl=1 ER - TY - JFULL T1 - Management of patients presenting with Sjogren's syndrome. A1 - Venables, PJ J1 - Best Pract Res Clin Rheumatol Y1 - 2006/08// VL - 20 SN - 1521-6942 SP - 791 EP - 807 N2 - Sjogren's syndrome is an autoimmune exocrinopathy that predominantly affects salivary and lachrymal glands, leading to dry eyes and mouth. The most common clinical problems faced by the rheumatologist are those of dry eyes and mouth, parotid swelling, fatigue and extraglandular manifestations. The first stage in management is to make an accurate diagnosis based on the American/European consensus criteria. The most frequent differential diagnoses are dry eyes and mouth symptoms, a variant of chronic fatigue syndrome and fibromyalgia, and sialosis, which causes a non-inflammatory enlargement of the parotid glands. The mainstay of treatment for the sicca symptoms is local therapy, and that for the milder systemic symptoms is hydroxychloroquine. Steroids and immunosuppressive drugs are reserved for more severe extraglandular disease. In spite of intensive research in other systemic treatments including biologic therapies, there is limited evidence to support their use in routine clinical practice. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16979538&query_hl=1 ER - TY - JFULL T1 - Expansion of zeta-dim CD4+ T-cells and zeta-dim natural-killer (NK)-cells correlates with C-reactive protein (CRP) in patients with acute coronary syndromes (ACS) A1 - Ammirati, E A1 - Banfi, M A1 - Cianflone, D A1 - Cope, AP A1 - Godino, C A1 - Mussardo, M A1 - Maseri, A A1 - Monaco, C J1 - EUR HEART J Y1 - 2006/08// VL - 27 SN - 0195-668X SP - 971 EP - 971 ER - TY - JFULL T1 - The mammalian chitinase-like lectin, YKL-40, binds specifically to type I collagen and modulates the rate of type I collagen fibril formation. A1 - Bigg, HF A1 - Wait, R A1 - Rowan, AD A1 - Cawston, TE J1 - J Biol Chem Y1 - 2006/07/28/ VL - 281 SN - 0021-9258 SP - 21082 EP - 21095 N2 - YKL-40 is expressed in arthritic cartilage and produced in large amounts by cultured chondrocytes, but its exact role is unclear, and the identities of its physiological ligands remain unknown. Purification of YKL-40 from resorbing bovine nasal cartilage and chondrocyte monolayers demonstrated the existence of three isoforms, a major and minor form from resorbing cartilage and a third species from chondrocytes. Affinity chromatography experiments with purified YKL-40 demonstrated specific binding of all three forms to collagen types I, II, and III, thus identifying collagens as potential YKL-40 ligands. Binding to immobilized type I collagen was inhibited by soluble native ligand, but not heat-denatured ligand, confirming a specific interaction. Binding of the chondrocyte-derived species to type I collagen was also demonstrated by surface plasmon resonance analysis, and the dissociation rate constant was calculated (3.42 x 10(-3) to 4.50 x 10(-3) s(-1)). The chondrocyte-derived species was found to prevent collagenolytic cleavage of type I collagen and to stimulate the rate of type I collagen fibril formation in a concentration-dependent manner. By contrast, the cartilage major form had an inhibitory effect on type I collagen fibrillogenesis. Digestion with N-glycosidase F, endoglycosidase H and lectin blotting did not reveal any difference in the carbohydrate component of these two YKL-40 species, indicating that this does not account for the opposing effects on fibril formation rate. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16704970&query_hl=1 ER - TY - JFULL T1 - Characterization and clinical application of human CD34+ stem/progenitor cell populations mobilized into the blood by granulocyte colony-stimulating factor. A1 - Gordon, MY A1 - Levicar, N A1 - Pai, M A1 - Bachellier, P A1 - Dimarakis, I A1 - Al-Allaf, F A1 - M'Hamdi, H A1 - Thalji, T A1 - Welsh, JP A1 - Marley, SB A1 - Davies, J A1 - Dazzi, F A1 - Marelli-Berg, F A1 - Tait, P A1 - Playford, R A1 - Jiao, L A1 - Jensen, S A1 - Nicholls, JP A1 - Ayav, A A1 - Nohandani, M A1 - Farzaneh, F A1 - Gaken, J A1 - Dodge, R A1 - Alison, M A1 - Apperley, JF A1 - Lechler, R A1 - Habib, NA J1 - Stem Cells Y1 - 2006/07// VL - 24 SN - 1066-5099 SP - 1822 EP - 1830 N2 - A phase I study was performed to determine the safety and tolerability of injecting autologous CD34(+) cells into five patients with liver insufficiency. The study was based on the hypothesis that the CD34(+) cell population in granulocyte colony-stimulating factor (G-CSF)-mobilized blood contains a subpopulation of cells with the potential for regenerating damaged tissue. We separated a candidate CD34(+) stem cell population from the majority of the CD34(+) cells (99%) by adherence to tissue culture plastic. The adherent and nonadherent CD34(+) cells were distinct in morphology, immunophenotype, and gene expression profile. Reverse transcription-polymerase chain reaction-based gene expression analysis indicated that the adherent CD34(+) cells had the potential to express determinants consistent with liver, pancreas, heart, muscle, and nerve cell differentiation as well as hematopoiesis. Overall, the characteristics of the adherent CD34(+) cells identify them as a separate putative stem/progenitor cell population. In culture, they produced a population of cells exhibiting diverse morphologies and expressing genes corresponding to multiple tissue types. Encouraged by this evidence that the CD34(+) cell population contains cells with the potential to form hepatocyte-like cells, we gave G-CSF to five patients with liver insufficiency to mobilize their stem cells for collection by leukapheresis. Between 1 x 10(6) and 2 x 10(8) CD34(+) cells were injected into the portal vein (three patients) or hepatic artery (two patients). No complications or specific side effects related to the procedure were observed. Three of the five patients showed improvement in serum bilirubin and four of five in serum albumin. These observations warrant further clinical trials. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16556705&query_hl=1 ER - TY - JFULL T1 - Over-expression in Escherichia coli and characterization of two recombinant isoforms of human FAD synthetase. A1 - Brizio, C A1 - Galluccio, M A1 - Wait, R A1 - Torchetti, EM A1 - Bafunno, V A1 - Accardi, R A1 - Gianazza, E A1 - Indiveri, C A1 - Barile, M J1 - Biochem Biophys Res Commun Y1 - 2006/06/09/ VL - 344 SN - 0006-291X SP - 1008 EP - 1016 N2 - FAD synthetase (FADS) (EC 2.7.7.2) is a key enzyme in the metabolic pathway that converts riboflavin into the redox cofactor FAD. Two hypothetical human FADSs, which are the products of FLAD1 gene, were over-expressed in Escherichia coli and identified by ESI-MS/MS. Isoform 1 was over-expressed as a T7-tagged protein which had a molecular mass of 63kDa on SDS-PAGE. Isoform 2 was over-expressed as a 6-His-tagged fusion protein, carrying an extra 84 amino acids at the N-terminal with an apparent molecular mass of 60kDa on SDS-PAGE. It was purified near to homogeneity from the soluble cell fraction by one-step affinity chromatography. Both isoforms possessed FADS activity and had a strict requirement for MgCl(2), as demonstrated using both spectrophotometric and chromatographic methods. The purified recombinant isoform 2 showed a specific activity of 6.8+/-1.3nmol of FAD synthesized/min/mg protein and exhibited a K(M) value for FMN of 1.5+/-0.3microM. This is the first report on characterization of human FADS, and the first cloning and over-expression of FADS from an organism higher than yeast. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16643857&query_hl=1 ER - TY - JFULL T1 - Proteomic analysis of the anterior cingulate cortex in the major psychiatric disorders: Evidence for disease-associated changes. A1 - Beasley, CL A1 - Pennington, K A1 - Behan, A A1 - Wait, R A1 - Dunn, MJ A1 - Cotter, D J1 - Proteomics Y1 - 2006/06// VL - 6 SN - 1615-9853 SP - 3414 EP - 3425 N2 - Abnormalities of the anterior cingulate cortex have previously been described in schizophrenia, major depressive disorder and bipolar disorder. In this study 2-DE was performed followed by mass spectrometric sequencing to identify disease-specific protein changes within the anterior cingulate cortex in these psychiatric disorders. The 2-DE system comprised IPGs 4-7 and 6-9 in the first, IEF dimension and SDS-PAGE in the second dimension. Resultant protein spots were compared between control and disease groups. Statistical analysis indicated that 35 spots were differentially expressed in one or more groups. Proteins comprising 26 of these spots were identified by mass spectroscopy. These represented 19 distinct proteins; aconitate hydratase, malate dehydrogenase, fructose bisphosphate aldolase A, ATP synthase, succinyl CoA ketoacid transferase, carbonic anhydrase, alpha- and beta-tubulin, dihydropyrimidinase-related protein-1 and -2, neuronal protein 25, trypsin precursor, glutamate dehydrogenase, glutamine synthetase, sorcin, vacuolar ATPase, creatine kinase, albumin and guanine nucleotide binding protein beta subunit. All but three of these proteins have previously been associated with the major psychiatric disorders. These findings provide support for the view that cytoskeletal and mitochondrial dysfunction are important components of the neuropathology of the major psychiatric disorders. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16637010&query_hl=1 ER - TY - JFULL T1 - Inhibition of tissue factor signalling reduces production of pro-inflammatory cytokines in atheroma smooth muscle cells A1 - Persson, L A1 - Navin, T A1 - Gregan, S A1 - Davies, AH A1 - Feldmann, M A1 - Monaco, C J1 - ATHEROSCLEROSIS SUPP Y1 - 2006/06// VL - 7 SN - 1567-5688 SP - 92 EP - 93 ER - TY - JFULL T1 - Enrichment of Z-dim CD4+ T-cells correlates with C-reactive protein (CRP) in patients with acute coronary syndrome (ACS) A1 - Ammirati, E A1 - Banfi, M A1 - Cianflone, D A1 - Cope, AP A1 - Maseri, A A1 - Monaco, C J1 - ATHEROSCLEROSIS SUPP Y1 - 2006/06// VL - 7 SN - 1567-5688 SP - 86 EP - 86 ER - TY - JFULL T1 - Proteomic analysis of membrane microdomains derived from both failing and non-failing human hearts A1 - Brioschi, M A1 - Wait, R A1 - Polvani, G A1 - Mussoni, L A1 - Tremoli, E A1 - Banfi, C J1 - ATHEROSCLEROSIS SUPP Y1 - 2006/06// VL - 7 SN - 1567-5688 SP - 591 EP - 591 ER - TY - JFULL T1 - Proteome of endothelial cell-derived procoagulant microparticles A1 - Brioschi, M A1 - Wait, R A1 - Mussoni, L A1 - Tremoli, E A1 - Banfi, C J1 - ATHEROSCLEROSIS SUPP Y1 - 2006/06// VL - 7 SN - 1567-5688 SP - 88 EP - 88 ER - TY - JFULL T1 - Comparison of extraction procedures for proteome analysis of Streptococcus pneumoniae and a basic reference map. A1 - Encheva, V A1 - Gharbia, SE A1 - Wait, R A1 - Begum, S A1 - Shah, HN J1 - Proteomics Y1 - 2006/06// VL - 6 SN - 1615-9853 SP - 3306 EP - 3317 N2 - Streptococcus pneumoniae is an important human pathogen causing life-threatening invasive diseases such as pneumonia, meningitis and bacteraemia. Despite major advances in our understanding of pneumococcal mechanisms of pathogenicity obtained through genomic studies very little has been achieved on the characterisation of the proteome of this pathogen. The highly complex structure of its cell envelope particularly amongst the various capsular forms enables the cell to resist lysis by conventional mechanical methods. It is therefore highly desirable to develop a cellular lysis and protein solubilisation procedure that minimises protein losses and allows for maximum possible coverage of the proteome of S. pneumoniae. Here we have utilised various combinations of mechanical or enzymatic cell lysis with two protein solubilisation mixtures urea/CHAPS-based mixture or SDS/DTT-based mixture in order to achieve best quality protein profiles using two proteomic technologies surface-enhanced laser desorption ionisation (SELDI) TOF MS and 2-DE. While urea/CHAPS-based mixture combined with freeze/thawing provided enough material for good-quality SELDI TOF MS fingerprints, a combination of mechanical, enzymatic and chemical lysis was needed to be used to successfully extract the desired protein content for 2-DE analysis. The methods chosen were also assessed for reproducibility and tested on various capsular types of S. pneumoniae. As a result, good-quality and reproducible profiles were created using various ProteinChip arrays and more than 800 protein spots were separated on a single 2-D gel of S. pneumoniae. Twenty-five of the most abundant protein spots were identified using LC/MS/MS to create a reference map of S. pneumoniae. The proteins identified included glycolytic enzymes such as glyceraldehyde 3-phosphate dehydrogenase, phosphoglycerate kinase, enolase etc. Several fermentation enzymes were also present including two of the components of the arginine deiminase system. Proteins involved in protein synthesis, such as translation factors and ribosomal proteins, as well as several chaperone proteins were also identified. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16673439&query_hl=1 ER - TY - JFULL T1 - Basic fibroblast growth factor: an extracellular mechanotransducer in articular cartilage? A1 - Vincent, T A1 - Saklatvala, J J1 - Biochem Soc Trans Y1 - 2006/06// VL - 34 SN - 0300-5127 SP - 456 EP - 457 N2 - Mechanical stimuli are important signals in articular cartilage, but what mediates them is unknown. We have shown that extracellular-signal-regulated kinase was activated on cutting and loading articular cartilage, and deduced that this was due to the release of bFGF (basic fibroblast growth factor) from the tissue. bFGF was shown to be extracellular, and by immunohistochemistry, was present in the pericellular matrix of articular chondrocytes attached to the heparan sulphate proteoglycan perlecan. We propose a novel mechanotransduction model, whereby pericellular bFGF, a short distance from the cell surface, becomes available to the cell surface tyrosine kinase receptors when articular cartilage is loaded. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16709186&query_hl=1 ER - TY - JFULL T1 - The human muscle proteome in aging. A1 - Gelfi, C A1 - Vigano, A A1 - Ripamonti, M A1 - Pontoglio, A A1 - Begum, S A1 - Pellegrino, MA A1 - Grassi, B A1 - Bottinelli, R A1 - Wait, R A1 - Cerretelli, P J1 - J Proteome Res Y1 - 2006/06// VL - 5 SN - 1535-3893 SP - 1344 EP - 1353 N2 - The aim of the present study was to assess age-dependent changes of proteins in the vastus lateralis muscle of physically active elderly and young subjects by a combination of two-dimensional difference gel electrophoresis, SDS-PAGE and ESI-MS/MS. The differences observed in the elderly group included down-regulation of regulatory myosin light chains, particularly the phosphorylated isoforms, a higher proportion of myosin heavy chain isoforms 1 and 2A, and enhanced oxidative and reduced glycolytic capacity. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16739986&query_hl=1 ER - TY - JFULL T1 - Rac1 and RhoA as regulators of endothelial phenotype and barrier function in hypoxia-induced neonatal pulmonary hypertension A1 - Wojciak-Stothard, B A1 - Tsang, LYF A1 - Paleolog, E A1 - Hall, SM A1 - Haworth, SG J1 - AM J PHYSIOL-LUNG C Y1 - 2006/06// VL - 290 SN - 1040-0605 SP - L1173 EP - L1182 N2 - Hypoxia is a common cause of persistant pulmonary hypertension in the newborn (PPHN), a condition associated with endothelial dysfunction and abnormal pulmonary vascular remodeling. The GTPase RhoA has been implicated in the pathogenesis of PPHN, but its contribution to endothelial remodeling and function is not known. We studied pulmonary artery endothelial cells (PAECs) taken from piglets with chronic hypoxia-induced pulmonary hypertension and from healthy animals and analyzed the roles of Rho GTPases in the regulation of the endothelial phenotype and function under basal normoxic conditions, acute hypoxia, and reoxygenation. The activities of RhoA, Rac1, and Cdc42 were correlated with changes in the endothelial cytoskeleton, adherens junctions, permeability, ROS production, VEGF levels, and activities of transcription factors hypoxia-inducible factor (HIF)-1 alpha and NF-kappa B. Adenoviral gene transfer was used to express dominant-negative GTPases, kinase-dead p21-activated kinase (PAK)-1, and constitutively activated Rac1 in cells. PAECs from pulmonary hypertensive piglets had a stable abnormal phenotype with a sustained reduction in Rac1 activity and an increase in RhoA activity, which correlated with an increase in actin stress fiber formation, increased permeability, and a decrease in VEGF and ROS production. Cells from pulmonary hypertensive animals were still able to respond to acute hypoxia. They also showed high activities of HIF-1 alpha and NF-kappa B, likely to result from changes in the activities of Rho GTPases. Activation of Rac1 and its effector PAK-1 as well as inhibition of RhoA restored the abnormal phenotype and permeability of hypertensive PAECs to normal. ER - TY - JFULL T1 - An absence of reactive oxygen species improves the resolution of lung influenza infection. A1 - Snelgrove, RJ A1 - Edwards, L A1 - Rae, AJ A1 - Hussell, T J1 - Eur J Immunol Y1 - 2006/06// VL - 36 SN - 0014-2980 SP - 1364 EP - 1373 N2 - Three influenza virus pandemics occurred in the last century, in 1918 killing 40-50 million people. In the absence of strain-specific vaccines, with potential resistance to antivirals and the threat of an imminent pandemic, strategies that alleviate symptoms are a priority. Reactive oxygen species are potent antimicrobial agents but cause immunopathology when produced in excess. Mice lacking a functional phagocyte NADPH oxidase (Cybb tm1 mice) or treated with the metalloporphyrin antioxidant manganese (III) tetrakis (N-ethyl pyridinium-2-yl) porpyhrin (MnTE-2-PyP) show heightened inflammatory infiltrates in their airways in response to pulmonary influenza infection, with augmented macrophage populations and a Th1-skewed T cell infiltrate. Underlying this exuberant macrophage response was a significant reduction in apoptosis and down-regulation of the myeloid inhibitory molecule CD200. Both, Cybb tm1 and MnTE-2-PyP-treated mice exhibited a reduced influenza titer in the lung parenchyma. Inflammatory infiltrate into the lung parenchyma was markedly reduced and lung function significantly improved. Manipulation of the homeostatic control of myeloid cells by inflammatory mediators therefore represents a novel therapeutic strategy in the treatment of influenza virus infection. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16703568&query_hl=1 ER - TY - JFULL T1 - Serial measurement of BCR-ABL transcripts in the peripheral blood after allogeneic stem cell transplantation for chronic myeloid leukemia: an attempt to define patients who may not require further therapy. A1 - Kaeda, J A1 - O'Shea, D A1 - Szydlo, RM A1 - Olavarria, E A1 - Dazzi, F A1 - Marin, D A1 - Saunders, S A1 - Khorashad, JS A1 - Cross, NC A1 - Goldman, JM A1 - Apperley, JF J1 - Blood Y1 - 2006/05/15/ VL - 107 SN - 0006-4971 SP - 4171 EP - 4176 N2 - We identified 243 patients with Philadelphia (Ph) chromosome-positive chronic myeloid leukemia (CML) who had BCR-ABL transcripts monitored by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) after allogeneic stem cell transplantation for a median of 84.3 months. Individual patients were regarded as having achieved molecular relapse (MR) if the BCR-ABL/ABL ratio exceeded 0.02% on 3 occasions or reached 0.05% on 2 occasions. Patients were allocated to 1 of 4 categories: (1) 36 patients were "persistently negative" or had a single low-level positive result; (2) 51 patients, "fluctuating positive, low level," had more than 1 positive result but never more than 2 consecutive positive results; (3) 27 patients, "persistently positive, low level," had persisting low levels of BCR-ABL transcripts but never more than 3 consecutive positive results; and (4) 129 patients relapsed. In 107 of these, relapse was based initially only on molecular criteria; in 72 (67.3%) patients the leukemia progressed to cytogenetic or hematologic relapse either prior to or during treatment with donor lymphocyte infusions. We conclude that the pattern of BCR-ABL transcript levels after allograft is variable; only a minority of patients with fluctuating or persistent low levels of BCR-ABL transcripts satisfied our definitions of MR, whereas the majority of patients who did so were likely to progress further. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16449534&query_hl=1 ER - TY - JFULL T1 - Shedding of lymphocyte function-associated antigen-1 (LFA-1) in a human inflammatory response. A1 - Evans, BJ A1 - McDowall, A A1 - Taylor, PC A1 - Hogg, N A1 - Haskard, DO A1 - Landis, RC J1 - Blood Y1 - 2006/05/01/ VL - 107 SN - 0006-4971 SP - 3593 EP - 3599 N2 - Shedding of adhesion molecules has been described for members of the selectin and immunoglobulin superfamilies, but integrins are not known to be shed. Here, we describe shedding of the integrin lymphocyte function-associated antigen-1 (LFA-1; CD11a/CD18) from human leukocytes during the cutaneous inflammatory response to the blistering agent cantharidin. Expression of LFA-1 was significantly diminished on blister-infiltrated neutrophils (P < .001) and monocytes (P = .02) compared with cells in peripheral blood, but expression on lymphocytes remained unchanged. A capture enzyme-linked immunosorbent assay (ELISA) indicated that LFA-1 was shed into blister fluid as a heterodimer expressing an intact headpiece with I and I-like epitopes. However, a CD11a central region epitope, G25.2, was absent and this remained expressed as a "stub" on the cell surface of blister neutrophils. Western analysis of soluble LFA-1 revealed a truncated 110-kDa CD11a chain and a minimally truncated 86-kDa CD18 chain. However, LFA-1 was shed in a ligand-binding conformation, since it expressed KIM-127 and 24 activation epitopes and bound to solid-phase ICAM-1. Shed LFA-1 was also detected in a synovial effusion by ELISA and Western analysis. We hypothesize that LFA-1 shedding may play a role in leukocyte detachment after transendothelial migration and in regulating integrin-dependent outside-in signaling. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16418329&query_hl=1 ER - TY - JFULL T1 - Optimising ovine cerebrospinal fluid preparation for two-dimensional gel electrophoresis. A1 - Chen, RL A1 - Sage, EA A1 - Dunn, MJ A1 - Wait, R A1 - Preston, JE J1 - Proteomics Y1 - 2006/05// VL - 6 SN - 1615-9853 SP - 3170 EP - 3175 N2 - Biomarkers for neurodegenerative disorders are potentially present in cerebrospinal fluid (CSF) and can be detected using proteomic technologies. Since CSF is high in salt and low in protein, its study by proteomic methods requires appropriate sample preparation. In this study, we applied four different sample treatments to the same ovine CSF sample. Precipitation with acetone or using a 2-D Clean-Up Kit (GE Healthcare BioSciences, Little Chalfont, UK) preserved more proteins, and produced more gel spots than spin columns from Sigma and Bio-Rad. A 53-kDa spot, identified by MS/MS as transthyretin (TTR) tetramer, was not detected in samples treated with the 2-D Clean-Up Kit, though it was always present on all gels prepared using the other three methods. Western immunoblotting confirmed the low recovery of tetrameric TTR by the 2-D Clean-Up Kit and showed that the tetrameric form of TTR predominated in ovine but not in rat CSF. In one ovine CSF sample haemoglobin was found, indicating blood contamination. We conclude that acetone precipitation is a simple and efficient way to prepare ovine CSF for 2-DE. The use of the 2-D Clean-Up Kit leads to the disappearance of tetrameric TTR only from ovine CSF proteome. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16619304&query_hl=1 ER - TY - JFULL T1 - Context-specific functional effects of IFNGR1 promoter polymorphism. A1 - Koch, O A1 - Kwiatkowski, DP A1 - Udalova, IA J1 - Hum Mol Genet Y1 - 2006/05/01/ VL - 15 SN - 0964-6906 SP - 1475 EP - 1481 N2 - We report evidence of a polymorphism in the promoter region of IFNGR1 (encoding interferon-gamma receptor 1) that has opposite functional effects in different cellular contexts. It is a deletion/insertion polymorphism that is found in Africans but not Europeans or Asians, and has been associated with resistance to severe malaria. We find that the IFNGR1-470del allele acts to suppress binding of nuclear proteins to the IFNGR1 promoter region in a manner that is specific for cell type. In B-lymphocytes, the IFNGR1-470del allele suppresses the binding of a approximately 35 kDa nuclear protein and acts to increase reporter gene expression. In epithelial cells, the same allele acts to decrease gene expression and suppresses the binding of approximately 90 kDa STAT-1 and STAT-2 proteins. In T-lymphocytes, this allele causes only subtle differences in nuclear protein binding and has no significant effect on gene expression. These findings suggest a mechanism by which a single genetic variant may cause a broad range of phenotypic consequences. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16600993&query_hl=1 ER - TY - JFULL T1 - Targeting rheumatoid tenosynovial angiogenesis with cytokine inhibitors. A1 - Jain, A A1 - Kiriakidis, S A1 - Brennan, F A1 - Sandison, A A1 - Paleolog, E A1 - Nanchahal, J J1 - Clin Orthop Relat Res Y1 - 2006/05// VL - 446 SN - 0009-921X SP - 268 EP - 277 N2 - Proliferation and invasion of the tenosynovial lining of tendons in patients with rheumatoid arthritis can result in tendon damage and rupture, leading to decreased hand function. Angiogenesis is an important process in rheumatoid joint disease; however, the role of angiogenesis in tendon disease is unknown. Our aim was to determine whether rheumatoid tenosynovial lining could produce angiogenic proteins, and if inhibition of tumor necrosis factor-alpha and interleukin-1 could decrease vascular endothelial growth factor production. Samples of encapsulating and invasive tenosynovial lining taken from the same hand and wrist synovial lining were harvested from 58 patients with rheumatoid arthritis having wrist surgery. Ex vivo samples were studied to quantify vascularity, angiogenic protein production under normoxic and hypoxic conditions, and the effect of inhibiting tumor necrosis factor-alpha and interleukin-1 on vascular endothelial growth factor production. Rheumatoid tenosynovial lining was more vascular than rheumatoid joint synovial lining and produced high levels of angiogenic factors such as vascular endothelial growth factor, interleukin-1beta, fibroblast growth factor-2, and angiopoietin-2. Hypoxia induced an increase in production of vascular endothelial growth factor by ex vivo tenosynovial lining cells. Inhibition of the cytokines interleukin-1 and tumor necrosis factor-alpha effectively reduced vascular endothelial growth factor production by tenosynovial samples. Level of Evidence: Therapeutic study. Level II (Prospective comparative study). See the Guidelines for Authors for a complete description of levels of evidence. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16456308&query_hl=1 ER - TY - JFULL T1 - Differential chemokine expression following respiratory virus infection reflects Th1- or Th2-biased immunopathology. A1 - Culley, FJ A1 - Pennycook, AM A1 - Tregoning, JS A1 - Hussell, T A1 - Openshaw, PJ J1 - J Virol Y1 - 2006/05// VL - 80 SN - 0022-538X SP - 4521 EP - 4527 N2 - Respiratory syncytial virus (RSV) is a major viral pathogen of infants that also reinfects adults. During RSV infection, inflammatory host cell recruitment to the lung plays a central role in determining disease outcome. Chemokines mediate cell recruitment to sites of inflammation and are influenced by, and influence, the production of cytokines. We therefore compared chemokine production in a mouse model of immunopathogenic RSV infection in which either Th1 or Th2 immunopathology is induced by prior sensitization to individual RSV proteins. Chemokine expression profiles were profoundly affected by the nature of the pulmonary immunopathology: "Th2" immunopathology in BALB/c mice was associated with increased and prolonged expression of CCL2 (MCP-1), CXCL10 (IP-10), and CCL11 (eotaxin) starting within 24 h of challenge. C57BL/6 mice with "Th2" pathology (enabled by a deficiency of CD8+ cells) also showed increased CCL2 production. No differences in chemokine receptor expression were detected. Chemokine blockers may therefore be of use for children with bronchiolitis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16611912&query_hl=1 ER - TY - JFULL T1 - Is there a role for TNF-alpha in anti-neutrophil cytoplasmic antibody-associated vasculitis? Lessons from other chronic inflammatory diseases. A1 - Feldmann, M A1 - Pusey, CD J1 - J Am Soc Nephrol Y1 - 2006/05// VL - 17 SN - 1046-6673 SP - 1243 EP - 1252 N2 - Anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis is the most common cause of rapidly progressive glomerulonephritis and immune-mediated pulmonary renal syndrome. Now that the acute manifestations of the disease generally can be controlled with immunosuppressive drugs, ANCA-associated vasculitis has become a chronic and relapsing inflammatory disorder. The need to develop safer and more effective treatment has led to great interest in the mediators of chronic inflammation. There are many lessons to be learned from studies of other chronic inflammatory diseases, particularly rheumatoid arthritis (RA). The identification of a TNF-alpha-dependent cytokine cascade in the in vitro cultures of synovium in joints of patients with RA led to studies of TNF blockade in experimental models of arthritis and subsequently to clinical trials. These have culminated in the widespread introduction of anti-TNF therapy not only in RA but also in Crohn disease, ankylosing spondylitis, and several other chronic inflammatory disorders. Following a similar investigative pathway, studies that show the importance of TNF production by leukocytes and intrinsic renal cells in glomerulonephritis have been followed by the demonstration of the effectiveness of TNF blockade in several experimental models of glomerulonephritis and vasculitis. In experimental autoimmune vasculitis, improvement in disease was paralleled by a reduction in leukocyte transmigration, as demonstrated by intravital microscopy. The benefit of infliximab (a mAb to TNF) in ANCA-associated vasculitis was recently reported in a prospective open-label study. However, the use of etanercept (a soluble TNF receptor fusion protein) was not found to be of significant benefit in a randomized, controlled trial in patients with Wegener granulomatosis. Therefore, there is a need for further evaluation of the use of anti-TNF antibodies in patients with ANCA-associated glomerulonephritis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16624928&query_hl=1 ER - TY - JFULL T1 - Isolation and characterization of cotiaractivase, a novel low molecular weight prothrombin activator from the venom of Bothrops cotiara. A1 - Senis, YA A1 - Kim, PY A1 - Fuller, GL A1 - García, A A1 - Prabhakar, S A1 - Wilkinson, MC A1 - Brittan, H A1 - Zitzmann, N A1 - Wait, R A1 - Warrell, DA A1 - Watson, SP A1 - Kamiguti, AS A1 - Theakston, RD A1 - Nesheim, ME A1 - Laing, GD J1 - Biochim Biophys Acta Y1 - 2006/05// VL - 1764 SN - 0006-3002 SP - 863 EP - 871 N2 - In this study, we isolated a novel prothrombin activator from the venom of Bothrops cotiara, a Brazilian lance-headed pit viper (Cotiara, Jararaca preta, Biocotiara), which we have designated "cotiaractivase" (prefix: cotiar- from B. cotiara; suffix: -activase, from prothrombin activating activity). Cotiaractivase was purified using a phenyl-Superose hydrophobic interaction column followed by a Mono-Q anion exchange column. It is a single-chain polypeptide with a molecular weight of 22,931 Da as measured by mass spectroscopy. Cotiaractivase generated active alpha-thrombin from purified human prothrombin in a Ca2+-dependent manner as assessed by S2238 chromogenic substrate assay and SDS-PAGE. Cotiaractivase cleaved prothrombin at positions Arg271-Thr272 and Arg320-Ile321, which are also cleaved by factor Xa. However, the rate of thrombin generation by cotiaractivase was approximately 60-fold less than factor Xa alone and 17 x 10(6)-fold less than the prothrombinase complex. The enzymatic activity of cotiaractivase was inhibited by the chelating agent EDTA, whereas the serine protease inhibitor PMSF had no effect on its activity, suggesting that it is a metalloproteinase. Interestingly, S2238 inhibited cotiaractivase activity non-competitively, suggesting that this toxin contains an exosite that allows it to bind prothrombin independently of its active site. Tandem mass spectrometry and N-terminal sequencing of purified cotiaractivase identified peptides that were identical to regions of the cysteine-rich and disintegrin-like domains of known snake venom metalloproteinases. Cotiaractivase is a unique low molecular weight snake venom prothrombin activator that likely belongs to the metalloproteinase family of proteins. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16647309&query_hl=1 ER - TY - JFULL T1 - The role of mesenchymal stem cells in haemopoiesis. A1 - Dazzi, F A1 - Ramasamy, R A1 - Glennie, S A1 - Jones, SP A1 - Roberts, I J1 - Blood Rev Y1 - 2006/05// VL - 20 SN - 0268-960X SP - 161 EP - 171 N2 - The ontogeny of haemopoiesis during fetal life and the differentiation of blood cells in adult life depend upon a fully competent microenvironment to provide appropriate signals via production of soluble factors and cell contact interactions. The cellular constituents of the microenvironment, also defined as the haemopoietic niche, largely derive from a common progenitor of mesenchymal origin. Mesenchymal stem cells (MSC), initially identified in adult bone marrow, have also been described in fetal haemopoietic tissues where they accompany the migration of haemopoietic development. Their precise identity remains ill-defined because of the lack of specific markers. Their ability to self-renew and differentiate into tissues of mesodermal origin (osteocytes, adipocytes, chondrocytes) and their lack of expression of haemopoietic molecules are currently the main criteria for isolation. In the bone marrow the most important elements of the niche appear to be osteoblasts, whilst a less defined population of fibroblasts regulates the maturation of immature T cells in the thymus. Recently, MSC have been shown to exert a profound immunosuppressive effect on polyclonal as well as antigen-specific T cell responses by inducing a state of division arrest anergy. Thus, the multipotent capacity of MSC, their role in supporting haemopoiesis, and their immunoregulatory activity make MSC particularly attractive for therapeutic exploitation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16364518&query_hl=1 ER - TY - JFULL T1 - Cyclooxygenase-2 induction and prostacyclin release by protease-activated receptors in endothelial cells require cooperation between mitogen-activated protein kinase and NF-kappaB pathways. A1 - Syeda, F A1 - Grosjean, J A1 - Houliston, RA A1 - Keogh, RJ A1 - Carter, TD A1 - Paleolog, E A1 - Wheeler-Jones, CP J1 - J Biol Chem Y1 - 2006/04/28/ VL - 281 SN - 0021-9258 SP - 11792 EP - 11804 N2 - The functional significance of protease-activated receptors (PARs) in endothelial cells is largely undefined, and the intracellular consequences of their activation are poorly understood. Here, we show that the serine protease thrombin, a PAR-1-selective peptide (TFLLRN), and SLIGKV (PAR-2-selective peptide) induce cyclooxygenase-2 (COX-2) protein and mRNA expression in human endothelial cells without modifying COX-1 expression. COX-2 induction was accompanied by sustained production of 6-keto-PGF1alpha, the stable hydrolysis product of prostacyclin, and this was inhibited by indomethacin and the COX-2-selective inhibitor NS398. PAR-1 and PAR-2 stimulation rapidly activated both ERK1/2 and p38MAPK, and pharmacological blockade of MEK with either PD98059 or U0126 or of p38MAPK by SB203580 or SB202190 strongly inhibited thrombin- and SLIGKV-induced COX-2 expression and 6-keto-PGF1alpha formation. Thrombin and peptide agonists of PAR-1 and PAR-2 increased luciferase activity in human umbilical vein endothelial cells infected with an NF-kappaB-dependent luciferase reporter adenovirus, and this, as well as PAR-induced 6-keto-PGF1alpha synthesis, was inhibited by co-infection with adenovirus encoding wild-type or mutated (Y42F) IkappaBalpha. Thrombin- and SLIGKV-induced COX-2 expression and 6-keto-PGF1alpha generation were markedly attenuated by the NF-kappaB inhibitor PG490 and partially inhibited by the proteasome pathway inhibitor MG-132. Activation of PAR-1 or PAR-2 promoted nuclear translocation and phosphorylation of p65-NF-kappaB, and thrombin-induced but not PAR-2-induced p65-NF-kappaB phosphorylation was reduced by inhibition of MEK or p38MAPK. Activation of PAR-4 by AYPGKF increased phosphorylation of ERK1/2 and p38MAPK without modifying NF-kappaB activation or COX-2 induction. Our data show that PAR-1 and PAR-2, but not PAR-4, are coupled with COX-2 expression and sustained endothelial production of vasculoprotective prostacyclin by mechanisms that depend on ERK1/2, p38MAPK, and IkappaBalpha-dependent NF-kappaB activation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16467309&query_hl=1 ER - TY - JFULL T1 - Individual Timp deficiencies differentially impact pro-MMP-2 activation A1 - English, JL A1 - Kassiri, Z A1 - Koskivirta, I A1 - Atkinson, SJ A1 - Di Grappa, M A1 - Soloway, PD A1 - Nagase, H A1 - Vuorio, E A1 - Murphy, G A1 - Khokha, R J1 - J BIOL CHEM Y1 - 2006/04/14/ VL - 281 SN - 0021-9258 SP - 10337 EP - 10346 N2 - Membrane-type matrix metalloproteinases (MT-MMPs) have emerged as key enzymes in tumor cell biology. The importance of MT1-MMP, in particular, is highlighted by its ability to activate pro-MMP-2 at the cell surface through the formation of a trimolecular complex comprised of MT1-MMP/tissue inhibitor of metalloproteinase-2 (TIMP-2)/pro-MMP-2. TIMPs1-4 are physiological MMP inhibitors with distinct roles in the regulation of pro-MMP-2 processing. Here, we have shown that individual Timp deficiencies differentially affect MMP-2 processing using primary mouse embryonic fibroblasts (MEFs). Timp-3 deficiency accelerated pro-MMP-2 activation in response to both cytochalasin D and concanavalin A. Exogenous TIMP-2 and N-TIMP-3 inhibited this activation, whereas TIMP-3 containing matrix from wild-type MEFs did not rescue the enhanced MMP-2 activation in Timp-3(-/-) cells. Increased processing of MMP-2 did not arise from increased expression of MT1-MMP, MT2-MMP, or MT3-MMP or altered expression of TIMP-2 and MMP-2. To test whether increased MMP-2 processing in Timp-3(-/-) MEFs is dependent on TIMP-2, double deficient Timp-2(-/-)/-3(-/-) MEFs were used. In these double deficient cells, the cleavage of pro-MMP-2 to its intermediate form was substantially increased, but the subsequent cleavage of intermediate-MMP-2 to fully active form, although absent in Timp-2(-/-) MEFs, was detectable with combined Timp-2(-/-)/-3(-/-) deficiency. TIMP-4 associates with MMP-2 and MT1-MMP in a manner similar to TIMP-3, but its deletion had no effect on pro-MMP-2 processing. Thus, TIMP-3 provides an inherent regulation over the kinetics of pro-MMP-2 processing, serving at a level distinct from that of TIMP-2 and TIMP-4. ER - TY - JFULL T1 - Reactivity of RA sera with citrullinated alpha-enolase peptides: Development of a new diagnostic ELISA A1 - Lundberg, K A1 - Kinloch, A A1 - Wait, R A1 - Moyes, D A1 - Svelander, L A1 - Klareskog, L A1 - Venables, P J1 - RHEUMATOLOGY Y1 - 2006/04// VL - 45 SN - 1462-0324 SP - I17 EP - I17 ER - TY - JFULL T1 - CD3Z polymorphism is associated with reduced expression of the TCR zeta chain: Enrichment in patients with severe rheumatoid arthritis A1 - Gorman, CL A1 - Russell, A A1 - Panesar, M A1 - Zhang, Z A1 - Vyse, TJ A1 - Cope, AP J1 - RHEUMATOLOGY Y1 - 2006/04// VL - 45 SN - 1462-0324 SP - I34 EP - I34 ER - TY - JFULL T1 - Sharp injury to articular cartilage activates JNK and NF kappa B and induces expression of inflammatory response genes A1 - Watt, FE A1 - Didangelos, A A1 - Sawaji, Y A1 - Saklatvala, J J1 - RHEUMATOLOGY Y1 - 2006/04// VL - 45 SN - 1462-0324 SP - I82 EP - I82 ER - TY - JFULL T1 - Post-streptococcal opsoclonus-myoclonus syndrome associated with anti-neuroleukin antibodies. A1 - Candler, PM A1 - Dale, RC A1 - Griffin, S A1 - Church, AJ A1 - Wait, R A1 - Chapman, MD A1 - Keir, G A1 - Giovannoni, G A1 - Rees, JH J1 - J Neurol Neurosurg Psychiatry Y1 - 2006/04// VL - 77 SN - 0022-3050 SP - 507 EP - 512 N2 - BACKGROUND: Adult opsoclonus-myoclonus (OM), a disorder of eye movements accompanied by myoclonus affecting the trunk, limbs, or head, is commonly associated with an underlying malignancy or precipitated by viral infection. METHODS: We present the first two reports of post-streptococcal OM associated with antibodies against a 56 kDa protein. Two young girls presented with opsoclonus and myoclonus following a febrile illness and pharyngitis. Protein purification techniques were employed. Amino acid sequences of human neuroleukin (NLK) and streptococcal proteins were compared using the protein-protein BLAST application. RESULTS: The antigen was identified as NLK (glucose-6-phosphate isomerase, GPI). GPI is present on the cell surface of streptococcus making the protein a candidate target for molecular mimicry. CONCLUSIONS: We have identified NLK as an antigenic target in two patients with post-streptococcal OM. The pathogenicity of the antibodies is uncertain. The potential role of anti-neuroleukin antibodies in the pathogenesis of OM is discussed. We propose that OM may represent a further syndrome in the growing spectrum of post-streptococcal neurological disorders. The role of streptococcus in OM and the frequency with which anti-NLK responses occur in both post-infectious and paraneoplastic OM should be investigated further. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16543530&query_hl=1 ER - TY - JFULL T1 - Hypoxia-driven angiogenesis is a key feature of tendon disease in rheumatoid arthritis A1 - Sivakumar, B A1 - Akhavani, M A1 - Kang, N A1 - Taylor, PC A1 - Paleolog, E J1 - RHEUMATOLOGY Y1 - 2006/04// VL - 45 SN - 1462-0324 SP - I39 EP - I40 ER - TY - JFULL T1 - The human heart proteome: Two-dimensional maps using narrow-range immobilised pH gradients. A1 - Westbrook, JA A1 - Wheeler, JX A1 - Wait, R A1 - Welson, SY A1 - Dunn, MJ J1 - Electrophoresis Y1 - 2006/04// VL - 27 SN - 0173-0835 SP - 1547 EP - 1555 N2 - The analysis of complex proteomes is undertaken using a variety of techniques and technologies such as 2-DE, surface-enhanced laser desorption ionisation, and various types of MS. In order to overcome the complexities of protein expression in discrete proteomes, sample fractionation has become an important aspect of proteomic experiments. The use of narrow-range IPGs (nrIPGs) is of special importance using the 2-DE proteomics workflow, since an enhanced visualisation of a given proteome is achieved through an improved physical separation and resolution of proteins. The work described in this paper presents a series of protein maps of the human heart left ventricle proteome that have been generated using nrIPGs for the first, IEF, dimension of 2-DE. A total of 374 gel spots were excised from seven different pH gradients, covering the range pH 3-10, giving rise to a total of 388 identifications from 110 unique proteins. Using Gene Ontologies (GOs), the identified proteins were found to be associated with 97 types of GO Process, 144 types of GO Function, and 54 types of GO Component. It is hoped that the maps presented in this paper will be of use to other researchers for reference purposes. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16609934&query_hl=1 ER - TY - JFULL T1 - Serum protein pattern during cow pregnancy: Acute-phase proteins increase in the peripartum period. A1 - Cairoli, F A1 - Battocchio, M A1 - Veronesi, MC A1 - Brambilla, D A1 - Conserva, F A1 - Eberini, I A1 - Wait, R A1 - Gianazza, E J1 - Electrophoresis Y1 - 2006/04// VL - 27 SN - 0173-0835 SP - 1617 EP - 1625 N2 - Serum collected in a time-course mode during the pregnancy of a group of heifers was analyzed by 2-DE under various experimental conditions to optimize resolution of all protein spots. Changes in the levels of some components were detected during the last phase of pregnancy and early postpartum. These included a decrease of alpha2-HS-glycoprotein, an increase of alpha1-antichymotrypsin and, with a much larger and more abrupt variation, of orosomucoid and haptoglobin. These findings associate the weeks preceding calving with an acute-phase reaction. Analysis of individual animal's sera by 1-DE demonstrated a higher level of orosomucoid in the sera of cows developing postpartum endometritis during the 2 wk after calving (i.e., in the course of the infection) but a lower level during the 2 wk before calving. This observation could represent an important tool for the prepartum detection of animals prone to develop postpartum endometritis and lead to a more accurate peripartum management of those animals. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16548048&query_hl=1 ER - TY - JFULL T1 - Citrullination is upregulated by pro-inflammatory cytokines: A new model for cytokine-driven autoimmunity in RA A1 - Moyes, DL A1 - Kinloch, A A1 - Lundberg, K A1 - Venables, PJ J1 - RHEUMATOLOGY Y1 - 2006/04// VL - 45 SN - 1462-0324 SP - I38 EP - I38 ER - TY - JFULL T1 - Localisation and characterisation of citrullinated proteins in the rheumatoid joint A1 - Kinloch, AJ A1 - Lundberg, K A1 - Wait, R A1 - Moyes, D A1 - Venables, PJ J1 - RHEUMATOLOGY Y1 - 2006/04// VL - 45 SN - 1462-0324 SP - I37 EP - I37 ER - TY - JFULL T1 - Boosting of cellular immunity against Mycobacterium tuberculosis and modulation of skin cytokine responses in healthy human volunteers by Mycobacterium bovis BCG substrain moreau Rio de Janeiro oral vaccine A1 - Cosgrove, CA A1 - Castello-Branco, LRR A1 - Hussell, T A1 - Sexton, A A1 - Giemza, R A1 - Phillips, R A1 - Williams, A A1 - Griffin, GE A1 - Dougan, G A1 - Lewis, DJM J1 - INFECT IMMUN Y1 - 2006/04// VL - 74 SN - 0019-9567 SP - 2449 EP - 2452 N2 - Oral immunization of healthy adults with 10(7) CFU BCG Moreau Rio de Janeiro was well tolerated and significantly boosted gamma interferon responses to purified protein derivative, Ag85, and MPB70 from previous childhood intradermal BCG immunization. Oral BCG offers the possibility of a needle-free tuberculosis vaccine and of boosting the protective immunity from intradermal tuberculosis vaccines. ER - TY - JFULL T1 - Modification of collagen-induced arthritis by education or manipulation of antigen presenting cells A1 - Edwards, L A1 - Snelgrove, R A1 - Feldmann, M A1 - Hussell, T J1 - RHEUMATOLOGY Y1 - 2006/04// VL - 45 SN - 1462-0324 SP - I18 EP - I18 ER - TY - JFULL T1 - A role for dendritic cells in the dissemination of mycobacterial infection. A1 - Humphreys, IR A1 - Stewart, GR A1 - Turner, DJ A1 - Patel, J A1 - Karamanou, D A1 - Snelgrove, RJ A1 - Young, DB J1 - Microbes Infect Y1 - 2006/04// VL - 8 SN - 1286-4579 SP - 1339 EP - 1346 N2 - The ability of mycobacteria to disseminate from the initial site of infection has an important role in immune priming and in the seeding of disease in multiple organs. To study this phenomenon, we used flow cytometry to analyse the distribution of green fluorescent protein-labelled BCG amongst different populations of antigen-presenting cells in the lungs of mice following intranasal infection, and monitored appearance of live bacteria in the draining mediastinal lymph nodes. BCG predominantly infected alveolar macrophages (CD11c(+)/CD11b(-)) and dendritic cells (CD11c(+)/CD11b(+)) in the lungs. The bacteria that disseminated to the lymph node were found in dendritic cells. The results are consistent with a model in which mycobacterial dissemination from the lung is initiated by the migration of infected dendritic cells to the draining lymph nodes. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16697232&query_hl=1 ER - TY - JFULL T1 - Proteolytic fragments of fibronectin containing the type III-13 and III-14 repeats induce aggrecanase activity in cartilage A1 - Sofat, N A1 - Gendron, CM A1 - Wait, R A1 - Enghild, JJ A1 - Nagase, H J1 - RHEUMATOLOGY Y1 - 2006/04// VL - 45 SN - 1462-0324 SP - I82 EP - I82 ER - TY - JFULL T1 - T cells expressing low levels of TCR xi preferentially migrate to sites of inflammation A1 - Gorman, CL A1 - Zhang, Z A1 - Monaco, C A1 - Marelli, F A1 - Schewitz, L A1 - Panesar, M A1 - Douglas, J A1 - McClinton, C A1 - Vallance, A A1 - Cope, AP J1 - RHEUMATOLOGY Y1 - 2006/04// VL - 45 SN - 1462-0324 SP - I17 EP - I18 ER - TY - JFULL T1 - Detection and properties of the human proliferative monocyte subpopulation. A1 - Clanchy, FI A1 - Holloway, AC A1 - Lari, R A1 - Cameron, PU A1 - Hamilton, JA J1 - J Leukoc Biol Y1 - 2006/04// VL - 79 SN - 0741-5400 SP - 757 EP - 766 N2 - Peripheral blood monocyte subpopulations have been reported and can give rise to diverse, differentiated phenotypes. A subpopulation(s) of human monocytes can proliferate in vitro in response to macrophage-colony stimulating factor (M-CSF; or CSF-1). This population, termed the proliferative monocyte (PM), is presumably less mature than other monocytes; however, it has not been defined further. Previous studies monitoring the frequency of the slowly cycling PM from different donors indicated that the assay for their reproducible measurement required improvement. We demonstrate that for optimal PM detection, high 5-bromo-2'-deoxyuridine concentrations are required over a delayed and wide time-frame. Surface marker phenotyping by flow cytometry showed that freshly isolated PM are CD14+ and could be distinguished from two other human monocyte subpopulations, namely, the CD14lo CD16+ and CD14lo CD64- subsets. PM express relatively high levels of CD64 and CD33 but have relatively low CD13 expression; they are also c-Fms+ and human leukocyte antigen-DR+. Labeling with carboxyfluorescein diacetate succinimidyl ester (CFSE) enabled the estimation of the number of PM divisions over time. Following CFSE labeling and culture, PM were sorted from the nonproliferating population and shown to have a distinctive, spindle-shaped morphology and higher capacity to form multinucleated, tartrate-resistant acid phosphatase+ cells in the presence of M-CSF and receptor activator of nuclear factor-kappaB ligand. The phenotype and properties of the PM subpopulation were examined as a prelude to determining its role in disease using methods that can be applied to clarify human monocyte heterogeneity. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16434691&query_hl=1 ER - TY - JFULL T1 - Bruton's tyrosine kinase is required for TLR2 and TLR4-induced TNF, but not IL-6, production. A1 - Horwood, NJ A1 - Page, TH A1 - McDaid, JP A1 - Palmer, CD A1 - Campbell, J A1 - Mahon, T A1 - Brennan, FM A1 - Webster, D A1 - Foxwell, BM J1 - J Immunol Y1 - 2006/03/15/ VL - 176 SN - 0022-1767 SP - 3635 EP - 3641 N2 - Bruton's tyrosine kinase (Btk), the gene mutated in the human immunodeficiency X-linked agammaglobulinemia, is activated by LPS and is required for LPS-induced TNF production. In this study, we have investigated the role of Btk both in signaling via another TLR (TLR2) and in the production of other proinflammatory cytokines such as IL-1beta, IL-6, and IL-8. Our data show that in X-linked agammaglobulinemia PBMCs, stimulation with TLR4 (LPS) or TLR2 (N-palmitoyl-S-[2, 3-bis(palmitoyloxy)-(2R)-propyl]-(R)-cysteine) ligands produces significantly less TNF and IL-1beta than in normal controls. In contrast, a lack of Btk has no impact on the production of IL-6, IL-8, or the anti-inflammatory cytokine, IL-10. Our previous data suggested that Btk lies within a p38-dependent pathway that stabilizes TNF mRNA. Accordingly, TaqMan quantitative PCR analysis of actinomycin D time courses presented in this work shows that overexpression of Btk is able to stabilize TNF, but not IL-6 mRNA. Furthermore, using the p38 inhibitor SB203580, we show that the TLR4-induced production of TNF, but not IL-6, requires the activity of p38 MAPK. These data provide evidence for a common requirement for Btk in TLR2- and TLR4-mediated induction of two important proinflammatory cytokines, TNF and IL-1beta, and reveal important differences in the TLR-mediated signals required for the production of IL-6, IL-8, and IL-10. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16517732&query_hl=1 ER - TY - JFULL T1 - Selective inhibition of matrix metalloproteinase-2 and-9 by phosphinic acid pseudo-peptides containing triple-helical structure A1 - Lauer-Fields, J A1 - Hammer, R A1 - Brew, K A1 - Nagase, H A1 - Fields, GB J1 - FASEB J Y1 - 2006/03/06/ VL - 20 SN - 0892-6638 SP - A48 EP - A48 ER - TY - JFULL T1 - Infliximab treatment maintains employability in patients with early rheumatoid arthritis. A1 - Smolen, JS A1 - Han, C A1 - van der Heijde, D A1 - Emery, P A1 - Bathon, JM A1 - Keystone, E A1 - Kalden, JR A1 - Schiff, M A1 - Bala, M A1 - Baker, D A1 - Han, J A1 - Maini, RN A1 - St Clair, EW J1 - Arthritis Rheum Y1 - 2006/03// VL - 54 SN - 0004-3591 SP - 716 EP - 722 N2 - OBJECTIVE: To evaluate the impact of infliximab therapy on the employment status of patients with early rheumatoid arthritis (RA). METHODS: Methotrexate (MTX)-naive patients with active early RA were randomly allocated to receive MTX plus placebo or MTX plus infliximab (3 mg/kg or 6 mg/kg) at weeks 0, 2, and 6 and then every 8 weeks through week 46. Data for patients younger than age 65 years were included in the analyses. A patient was categorized as employable if he or she was employed or felt well enough to work if a job were available. RESULTS: The change in actual employment was not significantly different between patients receiving MTX plus infliximab and those receiving MTX plus placebo (0.5% versus 1.3%; P > 0.5). However, the proportion of patients whose status changed from employable at baseline to unemployable at week 54 was smaller in the group receiving MTX plus infliximab compared with that in the group receiving MTX alone (8% versus 14%; P = 0.05). Patients who were treated with infliximab plus MTX had a significantly greater likelihood of improvement rather than deterioration in employability (odds ratio 2.4; P < 0.001); this likelihood was not significantly greater in patients receiving MTX alone. The proportion of employed patients who lost workdays during the trial was smaller in the MTX plus infliximab group than in the MTX-alone group (P = 0.010). CONCLUSION: The actual employment rates among patients in the 2 treatment groups were not different. However, patients with early RA who were treated with MTX plus infliximab had a higher probability of maintaining their employability compared with those who were treated with MTX alone. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16508932&query_hl=1 ER - TY - JFULL T1 - Posttranslational regulation of tristetraprolin subcellular localization and protein stability by p38 mitogen-activated protein kinase and extracellular signal-regulated kinase pathways. A1 - Brook, M A1 - Tchen, CR A1 - Santalucia, T A1 - McIlrath, J A1 - Arthur, JS A1 - Saklatvala, J A1 - Clark, AR J1 - Mol Cell Biol Y1 - 2006/03// VL - 26 SN - 0270-7306 SP - 2408 EP - 2418 N2 - The p38 mitogen-activated protein kinase (MAPK) signaling pathway, acting through the downstream kinase MK2, regulates the stability of many proinflammatory mRNAs that contain adenosine/uridine-rich elements (AREs). It is thought to do this by modulating the expression or activity of ARE-binding proteins that regulate mRNA turnover. MK2 phosphorylates the ARE-binding and mRNA-destabilizing protein tristetraprolin (TTP) at serines 52 and 178. Here we show that the p38 MAPK pathway regulates the subcellular localization and stability of TTP protein. A p38 MAPK inhibitor causes rapid dephosphorylation of TTP, relocalization from the cytoplasm to the nucleus, and degradation by the 20S/26S proteasome. Hence, continuous activity of the p38 MAPK pathway is required to maintain the phosphorylation status, cytoplasmic localization, and stability of TTP protein. The regulation of both subcellular localization and protein stability is dependent on MK2 and on the integrity of serines 52 and 178. Furthermore, the extracellular signal-regulated kinase (ERK) pathway synergizes with the p38 MAPK pathway to regulate both stability and localization of TTP. This effect is independent of kinases that are known to be synergistically activated by ERK and p38 MAPK. We present a model for the actions of TTP and the p38 MAPK pathway during distinct phases of the inflammatory response. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16508015&query_hl=1 ER - TY - JFULL T1 - Predictors of joint damage in patients with early rheumatoid arthritis treated with high-dose methotrexate with or without concomitant infliximab: results from the ASPIRE trial. A1 - Smolen, JS A1 - Van Der Heijde, DM A1 - St Clair, EW A1 - Emery, P A1 - Bathon, JM A1 - Keystone, E A1 - Maini, RN A1 - Kalden, JR A1 - Schiff, M A1 - Baker, D A1 - Han, C A1 - Han, J A1 - Bala, M A1 - Active-Controlled Study of Patients Receiving Infliximab for the Treatment of Rheumatoid Arthritis of Early Onset (ASPIRE) Study Group J1 - Arthritis Rheum Y1 - 2006/03// VL - 54 SN - 0004-3591 SP - 702 EP - 710 N2 - OBJECTIVE: To identify disease characteristics leading to progression of joint damage in patients with early rheumatoid arthritis (RA) treated with methotrexate (MTX) versus those treated with infliximab plus MTX. METHODS: Patients who had not previously been treated with MTX with active RA were randomly assigned to receive escalating doses of MTX up to 20 mg/week plus placebo or infliximab at weeks 0, 2, and 6, and every 8 weeks thereafter through week 46. Radiographic joint damage was assessed using the modified Sharp/van der Heijde score (SHS). The relationship between disease activity measures at baseline and week 14, as well as those averaged over time, were examined in relation to the change in SHS from baseline through week 54. RESULTS: C-reactive protein (CRP) levels, erythrocyte sedimentation rate (ESR), and swollen joint count were associated with greater joint damage progression in the MTX-only group, while none of these parameters was associated with progression in the infliximab plus MTX group. Mean changes in SHS among patients in the highest CRP (> or = 3 mg/dl) and ESR (> or = 52 mm/hour) tertiles in the MTX-only group were 5.62 and 5.89, respectively, compared with 0.73 and 1.12 in the infliximab plus MTX group (P < 0.001). Patients with greater joint damage at baseline (SHS > or = 10.5) showed less progression with infliximab plus MTX compared with MTX alone (-0.39 versus 4.11; P < 0.001). Patients receiving MTX alone who had persistently active disease at week 14 showed greater radiographic progression of joint damage than those taking MTX plus infliximab. CONCLUSION: High CRP level, high ESR, or persistent disease activity was associated with greater radiographic progression in the group taking MTX alone, while little radiographic progression was seen in patients receiving both MTX and infliximab, regardless of the abnormal levels of these traditional predictors. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16508926&query_hl=1 ER - TY - JFULL T1 - Correction of severe anaemia using immuno-regulated gene therapy is achieved by restoring the early erythroblast compartment. A1 - Du Roure, C A1 - Takács, K A1 - Maxwell, PH A1 - Roberts, I A1 - Dazzi, F A1 - Cannella, L A1 - Merkenschlager, M A1 - Fisher, AG J1 - Br J Haematol Y1 - 2006/03// VL - 132 SN - 0007-1048 SP - 608 EP - 614 N2 - Patients with chronic renal failure usually require exogenous erythropoietin (epo) to alleviate anaemia resulting from inadequate epo production by the kidneys. We have recently shown that severe anaemia in genetically manipulated epo-deficient mice (EpoTAg) can be corrected by adoptively transferred epo-producing lymphocytes. The aim of this study was to investigate the precise effects of human epo administration by this route on erythropoietic development in epo-deficient mice. The erythroblast compartments of untreated and treated EpoTAg mice were analysed in comparison with wild-type mice. The early erythroblast population was reduced in the bone marrow of epo-deficient mice, whilst the number of erythroid colony-forming units (CFU-E) was not significantly compromised. This paucity in marrow early erythroblasts was restored to normal values in treated mutant mice. In addition, the early erythroblast population was expanded in the spleens of treated animals. These findings show that the early erythroblasts are important targets of epo and that epo corrects anaemia of epo-deficient mice by restoring marrow function and splenic erythropoiesis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16445835&query_hl=1 ER - TY - JFULL T1 - Neuronal surface glycolytic enzymes are autoantigen targets in post-streptococcal autoimmune CNS disease. A1 - Dale, RC A1 - Candler, PM A1 - Church, AJ A1 - Wait, R A1 - Pocock, JM A1 - Giovannoni, G J1 - J Neuroimmunol Y1 - 2006/03// VL - 172 SN - 0165-5728 SP - 187 EP - 197 N2 - Infection with the Group A Streptococcus (GAS) can result in immune mediated brain disease characterised by a spectrum of movement and psychiatric disorders. We have previously described anti-neuronal antibodies in patients that bind to a restricted group of brain antigens with molecular weights 40 kDa, 45 kDa (doublet) and 60 kDa. The aim of this study was to define these antigens using 2-dimensional electrophoresis or ion exchange and hydrophobic interaction chromatography, followed by mass spectrometry. The findings were confirmed using commercial antibodies, commercial antigens and recombinant human antigens. The autoantigens were neuronal glycolytic enzymes--NGE (pyruvate kinase M1, aldolase C, neuronal-specific and non-neuronal enolase). These are multifunctional proteins that are all expressed intracellularly and on the neuronal cell surface. On the neuronal plasma membrane, NGE are involved in energy metabolism, cell signalling and synaptic neurotransmission. Anti-NGE antibodies were more common in the 20 unselected post-streptococcal CNS patients compared to 20 controls. In vitro experiments using cultured neurons showed that commercial anti-NGE antibodies induced apoptosis compared to blank incubation and control anti-HuD antibody. GAS also expresses glycolytic enzymes on cell surfaces that have 0-49% identity with human NGE, suggesting molecular mimicry and autoimmune cross-reactivity may be the pathogenic mechanism in post-streptococcal CNS disease. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16356555&query_hl=1 ER - TY - JFULL T1 - Coordinated and reversible reduction of enzymes involved in terminal oxidative metabolism in skeletal muscle mitochondria from a riboflavin-responsive, multiple acyl-CoA dehydrogenase deficiency patient. A1 - Gianazza, E A1 - Vergani, L A1 - Wait, R A1 - Brizio, C A1 - Brambilla, D A1 - Begum, S A1 - Giancaspero, TA A1 - Conserva, F A1 - Eberini, I A1 - Bufano, D A1 - Angelini, C A1 - Pegoraro, E A1 - Tramontano, A A1 - Barile, M J1 - Electrophoresis Y1 - 2006/03// VL - 27 SN - 0173-0835 SP - 1182 EP - 1198 N2 - In this case report we studied alterations in mitochondrial proteins in a patient suffering from recurrent profound muscle weakness, associated with ethylmalonic-adipic aciduria, who had benefited from high dose of riboflavin treatment. Morphological and biochemical alterations included muscle lipid accumulation, low muscle carnitine content, reduction in fatty acid beta-oxidation and reduced activity of complexes I and II of the respiratory chain. Riboflavin therapy partially or totally reversed these symptoms and increased the level of muscle flavin adenine dinucleotide, suggesting that aberrant flavin cofactor metabolism accounted for the disease. Proteomic investigation of muscle mitochondria revealed decrease or absence of several flavoenzymes, enzymes related to flavin cofactor-dependent mitochondrial pathways and mitochondrial or mitochondria-associated calcium-binding proteins. All these deficiencies were completely rescued after riboflavin treatment. This study indicates for the first time a profound involvement of riboflavin/flavin cofactors in modulating the level of a number of functionally coordinated polypeptides involved in fatty acyl-CoA and amino acid metabolism, extending the number of enzymatic pathways altered in riboflavin-responsive multiple acyl-CoA dehydrogenase deficiency. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16470778&query_hl=1 ER - TY - JFULL T1 - Amelioration of influenza-induced pathology in mice by coinfection with Trichinella spiralis. A1 - Furze, RC A1 - Hussell, T A1 - Selkirk, ME J1 - Infect Immun Y1 - 2006/03// VL - 74 SN - 0019-9567 SP - 1924 EP - 1932 N2 - Illness due to respiratory virus infection is often induced by excessive infiltration of cells into pulmonary tissues, leading to airway occlusion. We show here that infection with Trichinella spiralis results in lower levels of tumor necrosis factor in bronchoalveolar lavage fluid and inhibits cellular recruitment into the airways of mice coinfected with influenza A virus. Infiltration of neutrophils and CD4+ and CD8+ lymphocytes was reduced, resulting in animals gaining weight more rapidly following the initial phase of infection. Influenza resulted in a generalized increase in vascular permeability in pulmonary tissues, and this was suppressed by parasite infection, although the effects were restricted to the early phase of trichinosis. Moreover, the number of cells producing interleukin-10 (IL-10), and the local levels of this cytokine, were reduced, suggesting that amelioration of pulmonary pathology by parasite infection occurs independently of IL-10 production. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16495568&query_hl=1 ER - TY - JFULL T1 - Is abatacept an effective treatment for patients with RA who do not respond to other anti-TNF treatments? A1 - Taylor, PC J1 - Nat Clin Pract Rheumatol Y1 - 2006/03// VL - 2 SN - 1745-8382 SP - 128 EP - 129 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16932670&query_hl=1 ER - TY - JFULL T1 - Management of severe open tibial fractures: the need for combined orthopaedic and plastic surgical treatment in specialist centres. A1 - Naique, SB A1 - Pearse, M A1 - Nanchahal, J J1 - J Bone Joint Surg Br Y1 - 2006/03// VL - 88 SN - 0301-620X SP - 351 EP - 357 N2 - Although it is widely accepted that grade IIIB open tibial fractures require combined specialised orthopaedic and plastic surgery, the majority of patients in the UK initially present to local hospitals without access to specialised trauma facilities. The aim of this study was to compare the outcome of patients presenting directly to a specialist centre (primary group) with that of patients initially managed at local centres (tertiary group). We reviewed 73 consecutive grade IIIB open tibial shaft fractures with a mean follow-up of 14 months (8 to 48). There were 26 fractures in the primary and 47 in the tertiary group. The initial skeletal fixation required revision in 22 (47%) of the tertiary patients. Although there was no statistically-significant relationship between flap timing and flap failure, all the failures (6 of 63; 9.5%) occurred in the tertiary group. The overall mean time to union of 28 weeks was not influenced by the type of skeletal fixation. Deep infection occurred in 8.5% of patients, but there were no persistently infected fractures. The infection rate was not increased in those patients debrided more than six hours after injury. The limb salvage rate was 93%. The mean limb functional score was 74% of that of the normal limb. At review, 67% of patients had returned to employment, with a further 10% considering a return after rehabilitation. The times to union, infection rates and Enneking limb reconstruction scores were not statistically different between the primary and tertiary groups. The increased complications and revision surgery encountered in the tertiary group suggest that severe open tibial fractures should be referred directly to specialist centres for simultaneous combined management by orthopaedic and plastic surgeons. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16498010&query_hl=1 ER - TY - JFULL T1 - Mitogen-activated protein kinase-activated protein kinase 2 regulates tumor necrosis factor mRNA stability and translation mainly by altering tristetraprolin expression, stability, and binding to adenine/uridine-rich element A1 - Hitti, E A1 - Iakovleva, T A1 - Brook, M A1 - Deppenmeier, S A1 - Gruber, AD A1 - Radzioch, D A1 - Clark, AR A1 - Blackshear, PJ A1 - Kotlyarov, A A1 - Gaestel, M J1 - MOL CELL BIOL Y1 - 2006/03// VL - 26 SN - 0270-7306 SP - 2399 EP - 2407 N2 - The mitogen-activated protein kinase (MAPK) p38/MAPK-activated protein kinase 2 (MK2) signaling pathway plays an important role in the posttranscriptional regulation of tumor necrosis factor (TNF), which is dependent on the adenine/uridine-rich element (ARE) in the 3' untranslated region of TNF mRNA. After lipopolysaccharide (LPS) stimulation, MK2-deficient macrophages show a 90% reduction in TNF production compared to the wild type. Tristetraprolin (TTP), a protein induced by LPS, binds ARE and destabilizes TNF mRNA. Accordingly, macrophages lacking TTP produce large amounts of TNF. Here, we generated MK2/TTP double knockout mice and show that, after LPS stimulation, bone marrow-derived macrophages produce TNF mRNA and protein levels comparable to those of TTP knockout cells, indicating that in the regulation of TNF biosynthesis TTP is genetically downstream of MK2. In addition, we show that MK2 is essential for the stabilization of TTP mRNA, and phosphoryllation by MK2 leads to increased TTP protein stability but reduced A-RE affinity. These data suggest that MK2 inhibits the mRNA destabilizing activity of TTP and, in parallel, codegradation of TTP together, with the target mRNA resulting in increased cellular levels of TTP. ER - TY - JFULL T1 - Proteomic analysis of membrane microdomains derived from both failing and non-failing human hearts. A1 - Banfi, C A1 - Brioschi, M A1 - Wait, R A1 - Begum, S A1 - Gianazza, E A1 - Fratto, P A1 - Polvani, G A1 - Vitali, E A1 - Parolari, A A1 - Mussoni, L A1 - Tremoli, E J1 - Proteomics Y1 - 2006/03// VL - 6 SN - 1615-9853 SP - 1976 EP - 1988 N2 - Eukaryotic cells plasma membranes are organized into microdomains of specialized function such as lipid rafts and caveolae, with a specific lipid composition highly enriched in cholesterol and glycosphingolipids. In addition to their role in regulating signal transduction, multiple functions have been proposed, such as anchorage of receptors, trafficking of cholesterol, and regulation of permeability. However, an extensive understanding of their protein composition in human heart, both in failing and non-failing conditions, is not yet available. Membrane microdomains were isolated from left ventricular tissue of both failing (n = 15) and non-failing (n = 15) human hearts. Protein composition and differential protein expression was explored by comparing series of 2-D maps and subsequent identification by LC-MS/MS analysis. Data indicated that heart membrane microdomains are enriched in chaperones, cytoskeletal-associated proteins, enzymes and protein involved in signal transduction pathway. In addition, differential protein expression profile revealed that 30 proteins were specifically up- or down-regulated in human heart failure membrane microdomains. This study resulted in the identification of human heart membrane microdomain protein composition, which was not previously available. Moreover, it allowed the identification of multiple proteins whose expression is altered in heart failure, thus opening new perspectives to determine which role they may play in this disease. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16475230&query_hl=1 ER - TY - JFULL T1 - Vascular endothelial growth factor signalling in endothelial cell survival: a role for NFkappaB. A1 - Grosjean, J A1 - Kiriakidis, S A1 - Reilly, K A1 - Feldmann, M A1 - Paleolog, E J1 - Biochem Biophys Res Commun Y1 - 2006/02/17/ VL - 340 SN - 0006-291X SP - 984 EP - 994 N2 - Angiogenesis is the development of blood capillaries from pre-existing vessels. Vascular endothelial growth factor (VEGF) is a key regulator of vessel growth and regression, and acts as an endothelial survival factor by protecting endothelial cells from apoptosis. Many genes involved in cell proliferation and apoptosis are regulated by the nuclear factor kappa B (NFkappaB) transcription factor family. This study aimed to address the hypothesis that VEGF-mediated survival effects on endothelium involve NFkappaB. Using an NFkappaB-luciferase reporter adenovirus, we observed activation of NFkappaB following VEGF treatment of human umbilical vein endothelial cells. This was confirmed using electrophoretic mobility shift assay and found to involve nuclear translocation of NFkappaB sub-unit p65. However, NFkappaB activation occurred without degradation of inhibitory IkappaB proteins (IkappaBalpha, IkappaBbeta, and IkappaBepsilon). Instead, tyrosine phosphorylation of IkappaBalpha was observed following VEGF treatment, suggesting NFkappaB activation was mediated by degradation-independent dissociation of IkappaBalpha from NFkappaB. Adenovirus-mediated over-expression of either native IkappaBalpha, or of IkappaBalpha in which tyrosine residue 42 was mutated to phenylalanine, inhibited induction of NFkappaB-dependent luciferase activity in response to VEGF. Furthermore, VEGF-induced upregulation of mRNA for the anti-apoptotic protein Bcl-2 and cell survival following serum withdrawal was reduced following IkappaBalpha over-expression. This study highlights that different molecular mechanisms of NFkappaB activation may be involved downstream of stimuli which activate the endothelial lining of blood vessels. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16410078&query_hl=1 ER - TY - JFULL T1 - Structure and function of matrix metalloproteinases and TIMPs. A1 - Nagase, H A1 - Visse, R A1 - Murphy, G J1 - Cardiovasc Res Y1 - 2006/02/15/ VL - 69 SN - 0008-6363 SP - 562 EP - 573 N2 - Matrix metalloproteinases (MMPs), also called matrixins, function in the extracellular environment of cells and degrade both matrix and non-matrix proteins. They play central roles in morphogenesis, wound healing, tissue repair and remodelling in response to injury, e.g. after myocardial infarction, and in progression of diseases such as atheroma, arthritis, cancer and chronic tissue ulcers. They are multi-domain proteins and their activities are regulated by tissue inhibitors of metalloproteinases (TIMPs). This review introduces the members of the MMP family and discusses their domain structure and function, proenyme activation, the mechanism of inhibition by TIMPs and their significance in physiology and pathology. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16405877&query_hl=1 ER - TY - JFULL T1 - Pathogenspecific TLR signaling in mucosa: Mutual contribution of microbial TLR agonists and virulence factors A1 - Sirard, JC A1 - Bayard, M A1 - Didierlaurent, A J1 - EUR J IMMUNOL Y1 - 2006/02// VL - 36 SN - 0014-2980 SP - 260 EP - 263 N2 - Detection of microorganisms through microbe-associated molecular patterns (MAMP) by Toll-like receptors (TLR) is crucial to trigger protective immunity. In the mucosa, sentinel cells are exposed to MAMP from both pathogens and commensals; however, the TLR response is tightly controlled to avoid inflammation in response to commensals. Uropathogenic Escherichia coli (UPEC) trigger innate responses during urinary tract infection in a TLR4-dependent and CD14-independent manner. UPEC express virulence factors, such as type 1 fimbriae and/or P fimbriae, allowing bacterial attachment to the epithelium. in this issue of the European Journal of Immunology, Fisher et al. show that fimbriae are required to induce a TLR4-specific epithelial response. Depending on the fimbriae expressed by UPEC, different adaptor molecules are involved in TLR4 signaling. These data add to the recent body of evidence suggesting that TLR responses are regulated by co-receptors, such as receptors for virulence factors. In conclusion, the "pathogenic" TLR stimulation provides a novel way for the host to ignore commensal bacteria. ER - TY - JFULL T1 - Fibroblast-activation protein-alpha is expressed by chondrocytes following a proinflammatory stimulus and is elevated in osteoarthritis A1 - Milner, JM A1 - Kevorkian, L A1 - Young, DA A1 - Jones, D A1 - Wait, R A1 - Cravatt, B A1 - Donell, ST A1 - Clark, IM A1 - Rowan, AD A1 - Cawston, TE J1 - INT J EXP PATHOL Y1 - 2006/02// VL - 87 SN - 0959-9673 SP - A54 EP - A54 ER - TY - JFULL T1 - The utility of N,N-biotinyl glutathione disulfide in the study of protein S-glutathiolation. A1 - Brennan, JP A1 - Miller, JI A1 - Fuller, W A1 - Wait, R A1 - Begum, S A1 - Dunn, MJ A1 - Eaton, P J1 - Mol Cell Proteomics Y1 - 2006/02// VL - 5 SN - 1535-9476 SP - 215 EP - 225 N2 - Glutathione disulfide (GSSG) accumulates in cells under an increased oxidant load, which occurs during neurohormonal or metabolic stimulation as well as in many disease states. Elevated GSSG promotes protein S-glutathiolation, a reversible post-translational modification, which can directly alter or regulate protein function. We developed novel strategies for the study of protein S-glutathiolation that involved the simple synthesis of N,N-biotinyl glutathione disulfide (biotin-GSSG). Biotin-GSSG treatment of cells mimics a defined component of oxidative stress, namely a shift in the glutathione redox couple to the oxidized disulfide state. This induces widespread protein S-glutathiolation, which was detected on non-reducing Western blots probed with streptavidin-horseradish peroxidase and imaged using confocal fluorescence microscopy and ExtrAvidin-FITC. S-Glutathiolated proteins were purified using streptavidin-agarose and identified using proteomic methods. We conclude that biotin-GSSG is a useful tool in the investigation of protein S-glutathiolation and offers significant advantages over conventional methods or antibody-based strategies. These novel approaches may find widespread utility in the study of disease or redox signaling models where GSSG accumulation occurs. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16223748&query_hl=1 ER - TY - JFULL T1 - Tollip regulates proinflammatory responses to interleukin-1 and lipopolysaccharide. A1 - Didierlaurent, A A1 - Brissoni, B A1 - Velin, D A1 - Aebi, N A1 - Tardivel, A A1 - Käslin, E A1 - Sirard, JC A1 - Angelov, G A1 - Tschopp, J A1 - Burns, K J1 - Mol Cell Biol Y1 - 2006/02// VL - 26 SN - 0270-7306 SP - 735 EP - 742 N2 - Activation of interleukin-1 (IL-1) receptor (IL-1R), Toll-like receptor 2 (TLR2), and TLR4 triggers NF-kappaB and mitogen-activated protein kinase (MAPK)-dependent signaling, thereby initiating immune responses. Tollip has been implicated as a negative regulator of NF-kappaB signaling triggered by these receptors in in vitro studies. Here, deficient mice were used to determine the physiological contribution of Tollip to immunity. NF-kappaB, as well as MAPK, signaling appeared normal in Tollip-deficient cells stimulated with IL-1beta or the TLR4 ligand lipopolysaccharide (LPS). Similarly, IL-1beta- and TLR-driven activation of dendritic cells and lymphocytes was indistinguishable from wild-type cells. In contrast, the production of the proinflammatory cytokines, IL-6 and tumor necrosis factor alpha was significantly reduced after IL-1beta and LPS treatment at low doses but not at lethal doses of LPS. Tollip therefore controls the magnitude of inflammatory cytokine production in response to IL-1beta and LPS. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16428431&query_hl=1 ER - TY - JFULL T1 - Isolation of fibronectin fragments that induce aggrecan degradation in cartilage A1 - Sofat, N A1 - Enghild, J A1 - Nagase, H J1 - INT J EXP PATHOL Y1 - 2006/02// VL - 87 SN - 0959-9673 SP - A9 EP - A10 ER - TY - JFULL T1 - Colonic bacterial infection abrogates eosinophilic pulmonary disease. A1 - Williams, AE A1 - Edwards, L A1 - Hussell, T J1 - J Infect Dis Y1 - 2006/01/15/ VL - 193 SN - 0022-1899 SP - 223 EP - 230 N2 - Induction of immunity to one pathogen in the lungs modifies the microenvironment and alters immunopathological changes that result from a second, unrelated pulmonary infection. However, it is unclear whether immunity generated at distant sites also affects lung immune responses. Here, we show that infection with the gut-restricted bacterium Citrobacter rodentium modifies immunopathological changes that result from pulmonary Cryptococcus neoformans infection. Th2 cytokine-driven pulmonary eosinophilia induced by C. neoformans infection was reduced, and the enhanced Th1 cytokine environment afforded more-rapid clearance of the fungus in C. rodentium-immune mice. The activated and intraepithelial (CD103+) T cell populations that expand after C. neoformans infection were diminished in C. rodentium-immune mice. T cell cross-reactivity was absent, but cross-reactive antibodies were detected. It is of importance to the "hygiene hypothesis" that these data indicate that an immune response induced by a gut-restricted pathogen can modify the immune outcome after pulmonary infection, suggesting that cell-phenotype modifications occur across mucosal sites. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16362886&query_hl=1 ER - TY - JFULL T1 - An investigation of transmission ratio distortion in the central region of the human MHC. A1 - Hanchard, N A1 - Rockett, K A1 - Udalova, I A1 - Wilson, J A1 - Keating, B A1 - Koch, O A1 - Nijnik, A A1 - Diakite, M A1 - Herbert, M A1 - Kwiatkowski, D J1 - Genes Immun Y1 - 2006/01// VL - 7 SN - 1466-4879 SP - 51 EP - 58 N2 - Transmission ratio distortion (TRD) describes a significant departure from expected Mendelian inheritance ratios that is fundamental to both the biology of reproduction and statistical genetics. The relatively high fetal wastage in humans, with consequent selection of alleles in utero, makes it likely that TRD is prevalent in the human genome. The central region of the human major histocompatibility complex (MHC) is a strong TRD candidate, as it houses a number of immune and regulatory genes that may be important in pregnancy outcome. We used a nonhaplotype-based method to select 13 tagging SNPs from three central MHC candidate regions, and analysed their transmission in 380 newborns and their parents (1138 individuals). A TRD of 54:46 was noted in favour of the common allele of a promoter SNP in the CLIC1 gene (P = 0.025), with a similar distortion using haplotypes across the same gene region (P = 0.016). We also found evidence that markers in the CLIC1 gene region may have been subject to recent selection (P < 0.001). The study illustrates the potential benefits of screening for TRD and highlights the difficulties encountered therein. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16341054&query_hl=1 ER - TY - JFULL T1 - T-cell contact-dependent regulation of CC and CXC chemokine production in monocytes through differential involvement of NFkappaB: implications for rheumatoid arthritis. A1 - Beech, JT A1 - Andreakos, E A1 - Ciesielski, CJ A1 - Green, P A1 - Foxwell, BM A1 - Brennan, FM J1 - Arthritis Res Ther Y1 - 2006/// VL - 8 SN - 1478-6362 SP - R168 EP - R168 N2 - We and others have reported that rheumatoid arthritis (RA) synovial T cells can activate human monocytes/macrophages in a contact-dependent manner to induce the expression of inflammatory cytokines, including tumour necrosis factor alpha (TNFalpha). In the present study we demonstrate that RA synovial T cells without further activation can also induce monocyte CC and CXC chemokine production in a contact-dependent manner. The transcription factor NFkappaB is differentially involved in this process as CXC chemokines but not CC chemokines are inhibited after overexpression of IkappaBalpha, the natural inhibitor of NFkappaB. This effector function of RA synovial T cells is also shared by T cells activated with a cytokine cocktail containing IL-2, IL-6 and TNFalpha, but not T cells activated by anti-CD3 cross-linking that mimics TCR engagement. This study demonstrates for the first time that RA synovial T cells as well as cytokine-activated T cells are able to induce monocyte chemokine production in a contact-dependent manner and through NFkappaB-dependent and NFkappaB-independent mechanisms, in a process influenced by the phosphatidyl-inositol-3-kinase pathway. Moreover, this study provides further evidence that cytokine-activated T cells share aspects of their effector function with RA synovial T cells and that their targeting in the clinic has therapeutic potential. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17101049&query_hl=1 ER - TY - JFULL T1 - Ultrasonographic and radiographic results from a two-year controlled trial of immediate or one-year-delayed addition of infliximab to ongoing methotrexate therapy in patients with erosive early rheumatoid arthritis. A1 - Taylor, PC A1 - Steuer, A A1 - Gruber, J A1 - McClinton, C A1 - Cosgrove, DO A1 - Blomley, MJ A1 - Marsters, PA A1 - Wagner, CL A1 - Maini, RN J1 - Arthritis Rheum Y1 - 2006/01// VL - 54 SN - 0004-3591 SP - 47 EP - 53 N2 - OBJECTIVE: To compare the impact of immediate and delayed introduction of anti-tumor necrosis factor therapy on inflammation and structural damage in methotrexate (MTX)-treated patients with early rheumatoid arthritis (RA). METHODS: Twenty-four patients with erosive early RA (duration < 3 years) who were receiving MTX were randomized to receive infliximab 5 mg/kg or placebo infusions at weeks 0, 2, and 6, and then every 8 weeks through week 46. Beginning at week 54 and thereafter, all patients received infliximab 5 mg/kg. Metacarpophalangeal joints were scanned using high-frequency ultrasonography and power Doppler imaging. Radiographs were evaluated using the modified Sharp/van der Heijde scoring system. RESULTS: From baseline to week 54, total synovial thickness was significantly improved in the infliximab + MTX group compared with the placebo + MTX group (median reduction 95.8% versus 37.5%; P = 0.005), as was the total color Doppler area (CDA; vascularity assessment) (median reduction 100% and 47.1%, respectively; P = 0.025). From week 0 to week 110, no significant between-group difference was observed in the change from baseline for total synovial thickening or the total CDA. At week 54, greater progression in the Sharp/van der Heijde score was apparent in patients receiving placebo + MTX compared with those receiving infliximab + MTX. Although radiographic progression in the placebo + MTX group was greatly reduced in the second year (after initiation of infliximab therapy), marked differences were observed between the infliximab + MTX group (median change in the Sharp/van der Heijde score 4.0) and the placebo + MTX group (median change 14.5) from baseline to week 110 (P = 0.076). CONCLUSION: The results indicate that the efficacy of 2 years of combination therapy with infliximab + MTX for inhibiting cumulative structural damage was superior to that of 1 year of treatment with MTX alone followed by the addition of infliximab. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16385521&query_hl=1 ER - TY - JFULL T1 - The importance of T cell interactions with macrophages in rheumatoid cytokine production. A1 - Brennan, FM A1 - Foey, AD A1 - Feldmann, M J1 - Curr Top Microbiol Immunol Y1 - 2006/// VL - 305 SN - 0070-217X SP - 177 EP - 194 N2 - The analysis of suppression of cytokines in rheumatoid synovial tissue and fluid pioneered the studies of human cytokines in diseased tissue due to the relative ease of staining samples, even at the height of the inflammatory process. These studies led to the study of synovial cytokine regulation, and the identification of TNF as a therapeutic target, which has been amply validated in clinical trials and now routine therapy. The next key question was how is TNF disregulated in synovium. Are there differences between the mechanisms of synovial TNF production compared to the production of protective TNF during an immune response? Are there differences between the induction of the pro-inflammatory TNF and the anti inflammatory IL-10? The analysis of the interaction of the two most abundant synovial cells, T lymphocytes and macrophages has provided interesting clues to new therapeutic approaches based on disrupting T-macrophage interaction. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16724806&query_hl=1 ER - TY - JFULL T1 - Bruton's tyrosine kinase is required for TLR2 and TLR4-induced TNF, but not IL-6, production A1 - Horwood, NJ A1 - Page, TH A1 - McDaid, JP A1 - Palmer, CD A1 - Campbell, J A1 - Mahon, T A1 - Brennan, FM A1 - Webster, D A1 - Foxwell, BM J1 - J Immunol Y1 - 2006/// IS - 6 VL - 176 SN - 0022-1767 SP - 3635 EP - 3641 ER - TY - JFULL T1 - Mixed connective tissue disease. A1 - Venables, PJ J1 - Lupus Y1 - 2006/// VL - 15 SN - 0961-2033 SP - 132 EP - 137 N2 - Mixed connective tissue disease (MCTD) was first described in 1972 as a disease syndrome with overlapping features of systemic sclerosis, systemic lupus erythematosus (SLE) and polymyositis associated with antibodies to RNAse sensitive extractable nuclear antigen. When the antigen was subsequently characterized as polypeptides on the U1 ribonuclear protein component of the splicesosome (U1RNP), MCTD became the first rheumatic disease syndrome to be defined by a serologic test. Clinical features include a high frequency of Raynaud's syndrome, swollen hands, sclerodactyly, arthritis, polymyositis and interstitial lung disease. Over the last 30 years there has been a continuing debate as to whether MCTD constitutes a 'distinct clinical entity'. Here, I will review the pathological, immunogenetic and clinical features of MCTD and conclude that the debate remains unresolved. The early misconception that it has a relatively good prognosis has not stood the test of time with long-term follow-up studies. These have identified a tendency for MCTD to evolve into SLE or systemic sclerosis and highlighted pulmonary hypertension and scleroderma renal crisis as important causes of death. Providing it is realized that our appreciation of the clinical features associated with anti-U1RNP have evolved over time, MCTD remains a useful concept in clinical practice. Whether it can be credited with the term 'disease' awaits the demonstration of common etiopathological events underlying the development of antibodies to U1 RNP and their associated clinical features. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16634365&query_hl=1 ER - TY - JFULL T1 - Study of load balancing strategies for finite element computations on heterogeneous clusters A1 - Munasinghe, K A1 - Wait, R J1 - LECT NOTES COMPUT SC Y1 - 2006/// VL - 3732 SN - 0302-9743 SP - 1123 EP - 1130 N2 - We study strategies for redistributing the load in an adaptive finite clement computation performed oil a cluster of workstations. The cluster is assumed to be a heterogeneous, multi-user computing environment. The performance of a particular processor depends on both static factors, such as the processor hardware and dynamic factors, such as the system load and the work of other users.On a network, it is assumed that all processors are connected, but the topology of the finite element sub-domains can be interpreted as a processor topology and hence for each processor, it is possible to define set of neighbours. In finite element analysis, the quantity of computation on a processor is proportional to the size of the sub-domain plus some contribution from the neighbours. We consider schemes that modify the sub-domains by, in general, moving data to adjacent processors. The numerical experiments show the efficiency of the approach. ER - TY - JFULL T1 - Detection and properties of the human proliferative monocyte subpopulation. A1 - Clanchy, FI A1 - Holloway, AC A1 - Lari, R A1 - Cameron, PU A1 - Hamilton, JA J1 - J Leukoc Biol. Y1 - 2006/// VL - 79 SP - 757 EP - 766 ER - TY - JFULL T1 - Rheumatoid arthritis: a disease of chronic, low-amplitude signals transduced through T cell antigen receptors? A1 - Zhang, Z A1 - Gorman, C A1 - Clark, JM A1 - Cope, AP J1 - Wien Med Wochenschr Y1 - 2006/01// VL - 156 SN - 0043-5341 SP - 2 EP - 10 N2 - Technology has advanced to the stage where it is now possible to identify genes that confer low to moderate risk of developing autoimmune diseases such as rheumatoid arthritis (RA). This has been facilitated by the growing appreciation that these hard to detect genetic signals can only be defined in large cohorts of well characterized patients. In RA, the association between disease susceptibility and genes encoded within the MHC has been known for decades. Recent studies have identified several new candidate genes that provide further insights into the molecular nature of aberrant immune responses in chronic inflammatory diseases. Here, we describe some of these new genes. Based on their known functions we propose that in a subgroup of patients with RA inheritance of allelic variants at distinct loci could lead to dysregulation of adaptive immune responses characterized by chronic, low-amplitude signaling transduced by antigen T cell receptors. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16465609&query_hl=1 ER - TY - JFULL T1 - 2-D protein maps of rat gastrocnemius and soleus muscles: a tool for muscle plasticity assessment. A1 - Gelfi, C A1 - Viganò, A A1 - De Palma, S A1 - Ripamonti, M A1 - Begum, S A1 - Cerretelli, P A1 - Wait, R J1 - Proteomics Y1 - 2006/01// VL - 6 SN - 1615-9853 SP - 321 EP - 340 N2 - Functional characterization of muscle fibers relies on ATPase activity and on differential measurements of metabolic proteins, including mitochondrial and glycolytic enzymes, glucose, lactate and lactic acid transporters, calcium cycling proteins and components of the contractile machinery. The recent introduction of microarray technology has enabled detailed gene expression studies under different physiological and pathological conditions, thus generating novel hypotheses on muscle function. However, microarray approaches are limited by the incomplete genome coverage of currently available chips, and by poor correlation between mRNA concentration and protein expression level. We have used 2-DE and MS to build a reference map of proteins from rat mixed gastrocnemius and soleus muscle, and to assess qualitative and quantitative differences in protein distribution between these two functionally dissimilar muscles. More than 800 spots on each gel were detected by silver staining, of which 167 were excised, digested in-gel with trypsin and analyzed by ESI-MS/MS. One hundred and twenty eight distinct gene products were identified, including metabolic, transport and contractile proteins. Forty one spots displayed differences in relative expression level between mixed gastrocnemius and soleus samples. These data not only enable differentiation of functionally distinct slow-twitch and fast-twitch fiber types, but also provide tools for investigating muscle plasticity in response to physiological and environmental conditions such as aging or hypoxia. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16302281&query_hl=1 ER - TY - JFULL T1 - Angiogenesis as a therapeutic target in arthritis: lessons from oncology. A1 - Bainbridge, J A1 - Sivakumar, B A1 - Paleolog, E J1 - Curr Pharm Des Y1 - 2006/// VL - 12 SN - 1381-6128 SP - 2631 EP - 2644 N2 - Rheumatoid arthritis (RA) is a chronic disabling autoimmune inflammatory disease of unknown aetiology with a prevalence of about 1% in most parts of the world. As a result of the debilitating nature of the disease, sufferers struggle with the simple activities of daily living and frequently fail to remain in full time employment. Furthermore, the mortality associated with the disease is equivalent to that seen in triple vessel coronary artery disease. Over the 10-15 years, advances in understanding the mechanisms of RA pathogenesis based on studies of human cells and animal models of arthritis have led to the identification of new targets for therapeutic intervention. Despite these advances, a significant proportion of patients continue to exhibit disease which is refractory to such therapy. As an alternative to anti-cytokine therapy, formation of new blood vessels ('angiogenesis') represents a potentially attractive target for therapy in RA. Angiogenesis has been a putative target in cancer since it was first linked to tumour growth and metastases in the 1970s. A number of significant advances have been made in the development of anti-cancer therapy using such an approach. This review focuses on the potential for targeting angiogenesis in RA, building upon the experience of angiogenesis inhibition in the oncological setting. Through this we hope to emphasise the potential value of anti-angiogenic therapy in RA and identify future directions for optimising treatment of this disabling disease. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16842163&query_hl=1 ER - TY - JFULL T1 - Correction: Interleukin-6: a new therapeutic target. A1 - Smolen, JS A1 - Maini, RN J1 - Arthritis Res Ther Y1 - 2006/// VL - 8 SN - 1478-6362 SP - 407 N2 - L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=17049070&query_hl=1 ER - TY - JFULL T1 - Analysis of lipids from extremophilic bacteria A1 - da Costa, MS A1 - Nobre, MF A1 - Wait, R J1 - METHOD MICROBIOL Y1 - 2006/// VL - 35 SN - 0580-9517 SP - 127 EP - 159 ER - TY - JFULL T1 - MT1-MMP: a potent modifier of pericellular microenvironment. A1 - Itoh, Y A1 - Seiki, M J1 - J Cell Physiol Y1 - 2006/01// VL - 206 SN - 0021-9541 SP - 1 EP - 8 N2 - Cells are regulated by many different means, and there is more and more evidence emerging that changes in the microenvironment greatly affect cell function. MT1-MMP is a type I transmembrane proteinase which participates in pericellular proteolysis of extracellular matrix (ECM) macromolecules. The enzyme is cellular collagenase essential for skeletal development, cancer invasion, growth, and angiogenesis. MT1-MMP promotes cell invasion and motility by pericellular ECM degradation, shedding of CD44 and syndecan1, and by activating ERK. Thus MT1-MMP is one of the factors that influence the cellular microenvironment and thereby affect cell-signaling pathways and eventually alters cellular behavior. As a proteinase, MT1-MMP is regulated by inhibitors, but it also requires formation of a homo-oligomer complex, localization to migration front of the cells, and internalization to become a "functionally active" cell function modifier. Developing new means to inhibit "functional activity" of MT1-MMP may be a new direction to establish treatments for the diseases that MT1-MMP mediates such as cancer and rheumatoid arthritis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15920734&query_hl=1 ER - TY - JFULL T1 - Effects of oxidative stress on the cardiac myocyte proteome: modifications to peroxiredoxins and small heat shock proteins. A1 - Cullingford, TE A1 - Wait, R A1 - Clerk, A A1 - Sugden, PH J1 - J Mol Cell Cardiol Y1 - 2006/01// VL - 40 SN - 0022-2828 SP - 157 EP - 172 N2 - Endogenous oxidative stress is a likely cause of cardiac myocyte death in vivo. We examined the early (0-2 h) changes in the proteome of isolated cardiac myocytes from neonatal rats exposed to H2O2 (0.1 mM), focussing on proteins with apparent molecular masses of between 20 and 30 kDa. Proteins were separated by two-dimensional gel electrophoresis (2DGE), located by silver-staining and identified by mass spectrometry. Incorporation of [35S]methionine or 32Pi was also studied. For selected proteins, transcript abundance was examined by reverse transcriptase-polymerase chain reaction. Of the 38 protein spots in the region, 23 were identified. Two families showed changes in 2DGE migration or abundance with H2O2 treatment: the peroxiredoxins and two small heat shock protein (Hsp) family members: heat shock 27 kDa protein 1 (Hsp25) and alphaB-crystallin. Peroxiredoxins shifted to lower pI values and this was probably attributable to 'over-oxidation' of active site Cys-residues. Hsp25 also shifted to lower pI values but this was attributable to phosphorylation. alphaB-crystallin migration was unchanged but its abundance decreased. Transcripts encoding peroxiredoxins 2 and 5 increased significantly. In addition, 10 further proteins were identified. For two (glutathione S-transferase pi, translationally-controlled tumour protein), we could not find any previous references indicating their occurrence in cardiac myocytes. We conclude that exposure of cardiac myocytes to oxidative stress causes post-translational modification in two protein families involved in cytoprotection. These changes may be potentially useful diagnostically. In the short term, oxidative stress causes few detectable changes in global protein abundance as assessed by silver-staining. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16324708&query_hl=1 ER - TY - JFULL T1 - Internal standards in differentiating embryonic stem cells in vitro. A1 - Murphy, CL J1 - Methods Mol Biol Y1 - 2006/// VL - 329 SN - 1064-3745 SP - 101 EP - 112 N2 - Embryonic stem (ES) cell lines are important for use in developmental biology studies, and because these cells are totipotent, they may provide a much-needed source of differentiated cells for certain therapeutic applications. The phenotype of the ES cell in culture is often assessed by (semi)quantitative RNA analyses. In such cases, it is critical to use appropriate internal standards to correct for experimentally induced sources of error. This is particularly true for ES cell differentiation because it is heterogeneous in nature. We describe protocols for determining the suitability of housekeeping genes to act as internal controls in differentiating ES cell cultures. Such assessment is needed for every experimental condition under investigation. The protocol focuses on polymerase chain reaction; however, the principle and experimental design are applicable to any (semi)quantitative RNA assay. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16845987&query_hl=1 ER - TY - JFULL T1 - Fibroblast activation protein alpha is expressed by chondrocytes following a pro-inflammatory stimulus and is elevated in osteoarthritis. A1 - Milner, JM A1 - Kevorkian, L A1 - Young, DA A1 - Jones, D A1 - Wait, R A1 - Donell, ST A1 - Barksby, E A1 - Patterson, AM A1 - Middleton, J A1 - Cravatt, BF A1 - Clark, IM A1 - Rowan, AD A1 - Cawston, TE J1 - Arthritis Res Ther Y1 - 2006/// VL - 8 SN - 1478-6362 SP - R23 EP - R23 N2 - Arthritis is characterised by the proteolytic degradation of articular cartilage leading to a loss of joint function. Articular cartilage is composed of an extracellular matrix of proteoglycans and collagens. We have previously shown that serine proteinases are involved in the activation cascades leading to cartilage collagen degradation. The aim of this study was to use an active-site probe, biotinylated fluorophosphonate, to identify active serine proteinases present on the chondrocyte membrane after stimulation with the pro-inflammatory cytokines IL-1 and oncostatin M (OSM), agents that promote cartilage resorption. Fibroblast activation protein alpha (FAPalpha), a type II integral membrane serine proteinase, was identified on chondrocyte membranes stimulated with IL-1 and OSM. Real-time PCR analysis shows that FAPalpha gene expression is up-regulated by this cytokine combination in both isolated chondrocytes and cartilage explant cultures and is significantly higher in cartilage from OA patients compared to phenotypically normal articular cartilage. Immunohistochemistry analysis shows FAPalpha expression on chondrocytes in the superficial zone of OA cartilage tissues. This is the first report demonstrating the expression of active FAPalpha on the chondrocyte membrane and elevated levels in cartilage from OA patients. Its cell surface location and expression profile suggest that it may have an important pathological role in the cartilage turnover prevalent in arthritic diseases. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16507127&query_hl=1 ER - TY - JFULL T1 - Proteomic investigation of the molecular pathophysiology of dysferlinopathy. A1 - De Palma, S A1 - Morandi, L A1 - Mariani, E A1 - Begum, S A1 - Cerretelli, P A1 - Wait, R A1 - Gelfi, C J1 - Proteomics Y1 - 2006/01// VL - 6 SN - 1615-9853 SP - 379 EP - 385 N2 - Mutations in dysferlin gene cause several types of muscular dystrophy in humans, including the limb-girdle muscular dystrophy type 2B and the distal muscular dystrophy of Miyoshi. The dysferlin gene product is a membrane-associated protein belonging to the ferlins family of proteins. The function of the dysferlin protein and the cause of deterioration and regression of muscle fibres in its absence, are incompletely known. A functional clue may be the presence of six hydrophilic domains, C2, that bind calcium and mediate the interaction of proteins with cellular membranes. Dysferlin seems to be involved in the membrane fusion or repair. Molecular diagnosis of dysferlinopathies is now possible and the types of gene alterations that have been characterized so far include missense mutations, deletions and insertions. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16302276&query_hl=1 ER - TY - JFULL T1 - Efficacy of the MMP inhibitor MMI270 against lung metastasis following removal of orthotopically transplanted human colon cancer in rat A1 - Ogata, Y A1 - Matono, K A1 - Nakajima, M A1 - Sasatomi, T A1 - Mizobe, T A1 - Nagase, H A1 - Shirouzu, K J1 - INT J CANCER Y1 - 2006/01/01/ VL - 118 SN - 0020-7136 SP - 215 EP - 221 N2 - We have investigated the antitumor effects of synthetic MMP inhibitor MM1270 against postoperative lung metastasis from colon cancer in nude rat. The KM12SM human colon cancer cells were injected into the cecal wall, and at 5 weeks after the injection, the cecum was removed including the tumor. Then, 30 mg/kg of MM1270 was administered perorally twice per (lay for 2 or 4 weeks, either immediately after removal or after week 2 after the removal. At week 7 after the removal, lung metastasis was significantly inhibited by the early administration of MM1270 immediately after the tumor removal but not by the late administration. The survival rates were significantly higher in the rats treated by early administration of MM1270 compared to the survival rate in untreated rats. Moreover, no lung metastasis was detected in some rats with 24-weeks' survival treated by early administration. Lower microvessel density, lower PCNA Index and higher Apoptotic Index in the lung metastases of the rats treated with MM1270 were found compared to those in untreated rats. A beneficial effect of by early administration of MM1270 against postoperative lung metastases may be expected through inhibiting neovascularization of metastases in nude rat. (c) 2005 Wiley-Liss, Inc. ER - TY - JFULL T1 - Interleukin-6: a new therapeutic target. A1 - Smolen, JS A1 - Maini, RN J1 - Arthritis Res Ther Y1 - 2006/// VL - 8 Suppl 2 SN - 1478-6362 SP - S5 EP - S5 N2 - The therapeutic success of biological agents, especially the tumour necrosis factor (TNF) inhibitors, has opened a new chapter in the book of therapies for rheumatoid arthritis. Nevertheless, more than 50% of patients may not respond by > 50% improvement. New compounds have recently entered the treatment arena. One of these is rituximab, which depletes B cells, and another, abatacept, interferes with T-cell co-stimulation. However, although these agents may be effective in a number of patients who fail to respond to TNF blockade, they only rarely induce remission and overall 50% response rates do not exceed those with the TNF inhibitors. Among the major proinflammatory cytokines, IL-6 plays a pleiotropic role both in terms of activating the inflammatory response and osteoclastogenesis. Here, we review recent phase II trials of tocilizumab, a humanized anti-IL-6 receptor antibody that achieves a significant therapeutic response rate. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16899109&query_hl=1 ER - TY - JFULL T1 - Toll-like receptors: a new target in rheumatoid arthritis? A1 - Stefan K Drexler, Sandra M Sacre, Brian M Foxwell J1 - Expert Review of Clinical Immunology Y1 - 2006/// IS - 4 VL - 2 SP - 585 EP - 599 ER - TY - JFULL T1 - RSV infection provokes airway remodelling in non-sensitised allergen challenged mice A1 - Tourdot, S A1 - Hussell, T A1 - Openshaw, PJM A1 - Edwards, L A1 - Lloyd, CM J1 - IMMUNOLOGY Y1 - 2005/12// VL - 116 SN - 0019-2805 SP - 53 EP - 53 ER - TY - JFULL T1 - TLR5 signalling alters subsequent immune responses in the lung A1 - Didierlaurent, A A1 - Snelgrove, R A1 - Edwards, L A1 - Williams, A A1 - Sirard, JC A1 - Hussell, T J1 - IMMUNOLOGY Y1 - 2005/12// VL - 116 SN - 0019-2805 SP - 26 EP - 26 ER - TY - JFULL T1 - Homophilic complex formation of MT1-matrix metalloproteinase is essential to express collagenolytic activity on the cell surface A1 - Itoh, Y A1 - Ito, N A1 - Nagase, H A1 - Seiki, M J1 - INT J EXP PATHOL Y1 - 2005/12// VL - 86 SN - 0959-9673 SP - A77 EP - A77 ER - TY - JFULL T1 - Is there a critical role for MAPK p38 in disease development and chronicity of rheumatoid arthritis? A1 - Li, C A1 - Owen, S A1 - Green, P A1 - Amjadi, P A1 - Beech, J A1 - Brennan, FM J1 - IMMUNOLOGY Y1 - 2005/12// VL - 116 SN - 0019-2805 SP - 88 EP - 88 ER - TY - JFULL T1 - The efficacy of NF-kappa B inducing molecules as vaccine adjuvants A1 - Wales, J A1 - Andreakos, E A1 - Foxwell, B A1 - Williams, R A1 - Buchan, G A1 - Feldmann, M J1 - IMMUNOLOGY Y1 - 2005/12// VL - 116 SN - 0019-2805 SP - 108 EP - 108 ER - TY - JFULL T1 - Parameters affecting the expression and function of the late co-stimulatory molecule OX40 A1 - Gwyer, EL A1 - Edwards, L A1 - Snelgrove, RJ A1 - Nesbitt, A A1 - Shaw, S A1 - Hussell, T J1 - IMMUNOLOGY Y1 - 2005/12// VL - 116 SN - 0019-2805 SP - 7 EP - 7 ER - TY - JFULL T1 - CD200 governs compartmentalization and resolution of inflammatory responses to respiratory infection A1 - Snelgrove, RJ A1 - Edwards, L A1 - Barclay, AN A1 - Hussell, T J1 - IMMUNOLOGY Y1 - 2005/12// VL - 116 SN - 0019-2805 SP - 103 EP - 104 ER - TY - JFULL T1 - Structural basis for collagenolytic activity of matrix metalloproteinase-1 (collagenase 1) A1 - Visse, R A1 - Bode, W A1 - Maskos, K A1 - Nagase, H J1 - INT J EXP PATHOL Y1 - 2005/12// VL - 86 SN - 0959-9673 SP - A87 EP - A88 ER - TY - JFULL T1 - Management of chronic myeloid leukaemia in relapse following donor lymphocyte infusion induced remission: a retrospective study of the clinical trials committee of the British Society of Blood & Marrow Transplantation (BSBMT) A1 - Cummins, M A1 - Cwynarski, K A1 - Marktel, S A1 - Dazzi, F A1 - Cavenagh, J A1 - Clark, RE A1 - Holyoake, TL A1 - Milligan, D A1 - Parker, A A1 - Russell, NH A1 - Marks, DI J1 - BONE MARROW TRANSPL Y1 - 2005/12// VL - 36 SN - 0268-3369 SP - 1065 EP - 1069 N2 - Donor lymphocyte infusion (DLI) can restore remission in a high percentage of patients with chronic myeloid leukaemia (CML) who relapse after allogeneic stem cell transplant (SCT). Subsequent relapses after a DLI-induced remission do occur and the optimal management of these patients is not defined. A retrospective study of the practice of UK transplant centres was conducted. In all, 13 patients from seven centres were identified: all were treated for relapse post allogeneic SCT with DLI and achieved either a complete cytogenetic (n = 5) or molecular (n = 8) remission. All patients subsequently had a second relapse, at molecular (n = 7), cytogenetic (n = 4) and haematological (n = 2) levels. Further DLI was used in the treatment of 11 patients, imatinib mesylate in three and chemotherapy in two. The two patients with haematological relapse died of blastic disease. The remaining 11 patients achieved either a complete cytogenetic (n = 2) or molecular (n = 9) remission. Nine patients remain in molecular remission at a median follow-up of 29 months, seven of whom had received DLI alone as treatment for second relapse, one DLI plus imatinib and one imatinib alone. Toxicity following DLI for second relapse was low. Longer follow-up will be required to see if these second DLI-induced remissions will be durable. ER - TY - JFULL T1 - Radiographic benefit without clinical improvement in infliximab-treated patients with rheumatoid arthritis: comment on the article by Smolen et al - Reply A1 - Smolen, JS A1 - Maini, RN A1 - Han, CL A1 - Baker, D A1 - Lipsky, PL A1 - ATTRACT Study Grp J1 - ARTHRITIS RHEUM Y1 - 2005/12// VL - 52 SN - 0004-3591 SP - 4045 EP - 4047 ER - TY - JFULL T1 - A self-limiting colonic infection abrogates eosinophilic lung disease A1 - Williams, AE A1 - Edwards, L A1 - Hussell, T J1 - IMMUNOLOGY Y1 - 2005/12// VL - 116 SN - 0019-2805 SP - 21 EP - 21 ER - TY - JFULL T1 - Lung-marginated monocytes modulate pulmonary microvascular injury during early endotoxemia. A1 - O'Dea, KP A1 - Young, AJ A1 - Yamamoto, H A1 - Robotham, JL A1 - Brennan, FM A1 - Takata, M J1 - Am J Respir Crit Care Med Y1 - 2005/11/01/ VL - 172 SN - 1073-449X SP - 1119 EP - 1127 N2 - RATIONALE: The role of monocytes in acute endotoxemia has been ascribed to systemic release of mediators within the central circulation. Little is known about the potential role of "marginated" monocytes in regulating microvascular inflammatory signaling. OBJECTIVES: To investigate whether lung-marginated monocytes can locally activate pulmonary endothelial cells through cell contact-dependent interactions in early endotoxemia. METHODS: Mice were challenged with LPS to produce acute endotoxemia and pulmonary vascular injury. Adoptive transfer of ex vivo LPS-stimulated donor leukocytes to recipient mice was also performed to evaluate cell-associated inflammatory signaling between monocytes and endothelial cells within the lung. Cell suspensions from excised lungs were analyzed by flow cytometry for expression of tumor necrosis factor alpha (TNF-alpha) on monocytes and cell adhesion molecules on endothelial cells. RESULTS: Substantial numbers of monocytes rapidly marginated to the lungs after endotoxin challenge in mice, and lung-marginated monocytes expressed significantly higher levels of membrane TNF than circulating monocytes, due to higher TNF production by the marginated cells. Injection of activated wild-type donor leukocytes to wild-type or TNF receptor double knockout recipients demonstrated that lung-marginated monocytes can induce TNF-dependent upregulation of adhesion molecules on pulmonary endothelial cells. Injection of activated donor leukocytes from TNF knock-in mice that express uncleavable mutant membrane TNF also induced adhesion molecule upregulation in wild-type recipients without a systemic soluble TNF release. CONCLUSIONS: These results reveal a previously unacknowledged role for lung-marginated monocytes in early endotoxemia, exerting local, cell-associated TNF signaling within the pulmonary microcirculation, contributing to the evolution of pulmonary vascular injury. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16081546&query_hl=1 ER - TY - JFULL T1 - Planning your research training. A1 - Cope, AP A1 - Brennan, FM A1 - Hill Gaston, JS A1 - Haskard, DO J1 - Rheumatology (Oxford) Y1 - 2005/11// VL - 44 SN - 1462-0324 SP - 1339 EP - 1340 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16159949&query_hl=1 ER - TY - JFULL T1 - Reference maps of mouse serum acute-phase proteins: changes with LPS-induced inflammation and apolipoprotein A-I and A-II transgenes. A1 - Wait, R A1 - Chiesa, G A1 - Parolini, C A1 - Miller, I A1 - Begum, S A1 - Brambilla, D A1 - Galluccio, L A1 - Ballerio, R A1 - Eberini, I A1 - Gianazza, E J1 - Proteomics Y1 - 2005/11// VL - 5 SN - 1615-9853 SP - 4245 EP - 4253 N2 - We present reference maps of the mouse serum proteome (run under reducing and non-reducing conditions), from control animals, from mice injected with lipopolysaccharide (LPS) to induce systemic inflammation, and from mice transgenic for human apolipoproteins A-I and A-II. Seventy-seven spots/spot chains from the reducing gels were identified by HPLC MS/MS, representing 28 distinct proteins, including a species-specific protease inhibitor, contrapsin, and high levels of carboxylesterase. The concentrations of acute-phase reactants were monitored for 96 h after LPS challenge. The greatest changes (four-fold 48 h after LPS administration) were observed for haptoglobin and hemopexin. Orosomucoid/alpha(1)-acid glycoprotein and apolipoprotein A-I increased steadily, to 50-60% above baseline at 96 h from stimulation. In mice transgenic for human apolipoprotein A-I the levels of expression of orosomucoid/alpha(1)-acid glycoprotein, alpha(1)-macroglobulin, esterase, kininogen and contrapsin were altered compared to knockout mice lacking apolipoprotein A-I. In contrast, except for the presence of apolipoprotein A-II, no statistically significant difference was observed in mice transgenic for human apolipoprotein A-II. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16196095&query_hl=1 ER - TY - JFULL T1 - Proteomic dataset of mouse aortic smooth muscle cells. A1 - Mayr, U A1 - Mayr, M A1 - Yin, X A1 - Begum, S A1 - Tarelli, E A1 - Wait, R A1 - Xu, Q J1 - Proteomics Y1 - 2005/11// VL - 5 SN - 1615-9853 SP - 4546 EP - 4557 N2 - In an accompanying study (in this issue, DOI 10.1002/pmic.200402044), we have characterised the proteome of Sca-1(+) progenitor cells, which may function as precursors of vascular smooth muscle cells (SMCs). In the present study, we have analysed and mapped protein expression in aortic SMCs of mice, using 2-DE, MALDI-TOF MS and MS/MS. The 2-D system comprised a non-linear immobilised pH 3-10 gradient in the first dimension (separating proteins with pI values of pH 3-10), and 12%T SDS-PAGE in the second dimension (separating proteins in the range 15,000-150,000 Da). Of the 2400 spots visualised, a subset of 267 protein spots was analysed, with 235 protein spots being identified corresponding to 154 unique proteins. The data presented here are the first map of aortic SMCs and the most extensive analysis of SMC proteins published so far. This valuable tool should provide a basis for comparative studies of protein expression in vascular smooth muscle of transgenic mice and is available on our website hhtp://www.vascular-proteomics.com. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16240290&query_hl=1 ER - TY - JFULL T1 - Proteome of endothelial cell-derived procoagulant microparticles. A1 - Banfi, C A1 - Brioschi, M A1 - Wait, R A1 - Begum, S A1 - Gianazza, E A1 - Pirillo, A A1 - Mussoni, L A1 - Tremoli, E J1 - Proteomics Y1 - 2005/11// VL - 5 SN - 1615-9853 SP - 4443 EP - 4455 N2 - Microparticles (MP) are small membrane vesicles that are released from cells upon activation or during apoptosis. Cellular MP in body fluids constitute a heterogeneous population, differing in cellular origin, numbers, size, antigenic composition and functional properties. MP support coagulation by exposure of tissue factor (TF), the initiator of coagulation in vivo. Moreover, MP may transfer bioactive molecules to other cells, thereby stimulating them to produce cytokines, cell-adhesion molecules, growth factors and TF, and modulate endothelial functions. However, a comprehensive characterization of the antigenic composition of MP has been poorly defined. This study describes the protein composition of endothelial cell (EC)-derived MP (EMP) using a proteomic approach. MS analysis indicated the presence of newly described protein such as metabolic enzymes, proteins involved in adhesion and fusion processes, members of protein folding event, cytoskeleton associated proteins and nucleosome. In conclusion, circulating EMP behave as an actual storage pool, able to disseminate blood-borne TF activity and other bioactive effectors, as confirmed by our experiments showing an increased procoagulant activity of EC exposed to EMP. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16220532&query_hl=1 ER - TY - JFULL T1 - What does tumour necrosis factor excess do to the immune system long term? A1 - Clark, J A1 - Vagenas, P A1 - Panesar, M A1 - Cope, AP J1 - Ann Rheum Dis Y1 - 2005/11// VL - 64 Suppl 4 SN - 0003-4967 SP - iv70 EP - iv76 N2 - Members of the tumour necrosis factor (TNF)/TNF-receptor (TNF-R) superfamily coordinate the immune response at multiple levels. For example, TNF, LTalpha, LTbeta and RANKL provide signals required for lymphoid neogenesis, CD27, OX-40, 4-1BB and CD30 deliver costimulatory signals to augment immune responses, while pro-apoptotic members such as TNF, CD95L and TRAIL may contribute to the termination of the response. Biological identity of individual family members has been revealed through studies of gain of function or gene deficient mutants. Most notable are the development of spontaneous inflammatory polyarthritis in human TNF-globin transgenic mice, the auto-inflammatory syndromes resulting from mutations in the 55-kDa TNF-R, and, in particular, the obligatory role for the RANKL/RANK axis in osteoclastogenesis and bone remodelling. A growing appreciation of the molecular basis of signalling pathways transduced by TNF-R has provided a framework for better understanding the biology of this expanding family. For while the rapid and robust activation of NF-kappaB and MAPK pathways is typical of acute TNF-R engagement, the molecular basis of sustained receptor signalling remains a mystery, in spite of its relevance to chronic inflammatory and immune responses. Focusing on T cells, this report describes some of the molecular footprints of sustained TNF-R engagement and illustrates how these may influence immune function. A common theme arising is that prolonged TNF stimulation alters signalling thresholds over time. The authors propose that one major outcome of long term exposure to TNF is a state of localised IL-2 deficiency at sites of inflammation. The implications of this deficiency are discussed. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16239393&query_hl=1 ER - TY - JFULL T1 - The 2005 International Symposium on Advances in Targeted Therapies: what have we learned in the 2000s and where are we going? A1 - Maini, RN J1 - Ann Rheum Dis Y1 - 2005/11// VL - 64 Suppl 4 SN - 0003-4967 SP - iv106 EP - iv108 N2 - This essay summarises a personal perspective on where we have got to in the five years since the turn of the last century. The focus is on advances in knowledge of antitumour necrosis factor (anti-TNF) therapy of rheumatoid arthritis and other chronic immune-inflammatory rheumatic diseases. The accumulating knowledge is clarifying the scope and limitations of efficacy, safety, and durability of these agents. It is defining the unmet needs of patients and communities worldwide for the future. The call is for progress in providing more cost-effective advances in therapeutics that not only interrupt the disease process long term but also reverse the anatomical and functional consequences of disease and the impairment in quality of life. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16239377&query_hl=1 ER - TY - JFULL T1 - Proteomic dataset of Sca-1+ progenitor cells. A1 - Yin, X A1 - Mayr, M A1 - Xiao, Q A1 - Mayr, U A1 - Tarelli, E A1 - Wait, R A1 - Wang, W A1 - Xu, Q J1 - Proteomics Y1 - 2005/11// VL - 5 SN - 1615-9853 SP - 4533 EP - 4545 N2 - Embryonic stem cells (ES cells) can differentiate into endothelial cells and smooth muscle cells (SMCs), which participate in vascular angiogenesis. In this study, we differentiated mouse ES cells into Sca-1(+) cells, which have the potential to serve as vascular progenitor cells, and mapped their proteome by 2-DE using a pH 3-10 non-linear gradient and 12% SDS-polyacrylamide gels. A subset of 300 protein spots was analysed and mapped, with 241 protein spots being identified by their PMF using MALDI-TOF MS or by partial amino acid sequencing using MS/MS. Our protein map is the first of Sca-1(+) progenitor cells and will facilitate the identification of proteins differentially expressed during stem cell differentiation. The proteome of adult arterial SMCs is described in an accompanying paper (in this issue, DOI 10.1002/pmic.200402045). All data are made accessible on our website http://www.vascular-proteomics.com. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16240289&query_hl=1 ER - TY - JFULL T1 - Analysing the effect of novel therapies on cytokine expression in experimental arthritis. A1 - Williams, RO A1 - Inglis, JJ A1 - Simelyte, E A1 - Criado, G A1 - Sumariwalla, PF J1 - Int J Exp Pathol Y1 - 2005/10// VL - 86 SN - 0959-9673 SP - 267 EP - 278 N2 - Type II collagen-induced arthritis (CIA) is an animal model of rheumatoid arthritis that has been used extensively to address questions of disease pathogenesis and to validate novel therapeutic targets. Susceptibility to CIA is strongly associated with major histocompatibility complex class II genes, and the development of arthritis is accompanied by a robust T- and B-cell response to type II collagen. The main pathological features of CIA include proliferative synovitis with infiltration of inflammatory cells, pannus formation, cartilage degradation, erosion of bone and fibrosis. Pro-inflammatory cytokines, such as tumour necrosis factor alpha and interleukin-1beta, are expressed in the arthritic joints in both murine CIA and human rheumatoid arthritis, and blockade of these molecules results in amelioration of disease. Hence, there is a great deal of interest in the development of small-molecular-weight inhibitors of pro-inflammatory cytokines. There is also interest in the development and testing of drugs with the capacity to modulate the immune pathways involved in driving the inflammatory response in arthritis. For these reasons, there is a need to monitor the effect of novel treatments on cytokine expression in vivo. In this review, we outline the various techniques used to detect cytokines in experimental arthritis and describe how these techniques have been used to quantify changes in cytokine expression following therapeutic intervention. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16191099&query_hl=1 ER - TY - JFULL T1 - Structures of the O-linked oligosaccharides of a complex glycoconjugate from Pseudallescheria boydii. A1 - Pinto, MR A1 - Gorin, PA A1 - Wait, R A1 - Mulloy, B A1 - Barreto-Bergter, E J1 - Glycobiology Y1 - 2005/10// VL - 15 SN - 0959-6658 SP - 895 EP - 904 N2 - Nonreducing O-linked oligosaccharides were obtained from the peptidorhamnomannan of mycelia of Pseudallescheria boydii by alkaline beta-elimination under reducing conditions. They were separated by gel filtration chromatography to give three oligosaccharide fractions. The major oligosaccharide from fraction 1 was characterized by a combination of techniques including electrospray ionization quadrupole time-of-flight tandem mass spectrometry (ESI MS/MS), matrix-assisted laser desorption ionization mass spectrometry (MALDI MS), nuclear magnetic resonance (NMR), and methylation gas-liquid chromatography-mass spectrometry (GC-MS) analysis. It was branched, with a principal chain of alpha-Rhap-(1 --> 3)-alpha-Rhap-(1 --> 3)-alpha-Manp-(1 --> 2)-Man-ol substituted at O-6 of mannitol with an alpha-Glcp-(1 --> 4)-beta-Galp group. Species containing one and two additional alpha-Glcp-(1 --> 4) substituents in the rhamnose branch were also present. The major component of fraction 2 was a substructure of oligosaccharide-1, lacking a hexose from the Glc-Gal branch. Fraction 3 contained a mixture of smaller, unbranched, oligosaccharides. In hapten inhibition tests, fractions 1 and 2 blocked the reaction between peptidorhamnomannan (PRM) and rabbit anti-P. boydii mycelium hyperimmune serum by approximately 75%, whereas fraction 3 inhibited by approximately 55%. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15932920&query_hl=1 ER - TY - JFULL T1 - Reactive site mutations in tissue inhibitor of metalloproteinase-3 disrupt inhibition of matrix metalloproteinases but not tumor necrosis factor-alpha-converting enzyme. A1 - Wei, S A1 - Kashiwagi, M A1 - Kota, S A1 - Xie, Z A1 - Nagase, H A1 - Brew, K J1 - J Biol Chem Y1 - 2005/09/23/ VL - 280 SN - 0021-9258 SP - 32877 EP - 32882 N2 - Tissue inhibitor of metalloproteinase-3 (TIMP-3) is a dual inhibitor of the matrix metalloproteinases (MMPs) and some adamalysins, two families of extracellular and cell surface metalloproteinases that function in extracellular matrix turnover and the shedding of cell surface proteins. The mechanism of inhibition of MMPs by TIMPs has been well characterized, and since the catalytic domains of MMPs and adamalysins are homologous, it was assumed that the interaction of TIMP-3 with adamalysins is closely similar. Here we report that the inhibition of the extracellular region of ADAM-17 (tumor necrosis factor alpha-converting enzyme (TACE)) by the inhibitory domain of TIMP-3 (N-TIMP-3) shows positive cooperativity. Also, mutations in the core of the MMP interaction surface of N-TIMP-3 dramatically reduce the binding affinity for MMPs but have little effect on the inhibitory activity for TACE. These results suggest that the mechanism of inhibition of ADAM-17 by TIMP-3 may be distinct from that for MMPs. The mutant proteins are also effective inhibitors of tumor necrosis factor alpha (TNF-alpha) release from phorbol ester-stimulated cells, indicating that they provide a lead for engineering TACE-specific inhibitors that may reduce side effects arising from MMP inhibition and are possibly useful for treatment of diseases associated with excessive TNF-alpha levels such as rheumatoid arthritis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16079149&query_hl=1 ER - TY - JFULL T1 - An IL-7 fusion protein that shows increased thymopoietic ability. A1 - Henson, SM A1 - Snelgrove, R A1 - Hussell, T A1 - Wells, DJ A1 - Aspinall, R J1 - J Immunol Y1 - 2005/09/15/ VL - 175 SN - 0022-1767 SP - 4112 EP - 4118 N2 - The role of IL-7 during thymopoiesis has led to it being the focus of a number of therapeutic interventions. However, its small size and pleiotropic nature present problems for thymus-directed therapies. We have created a fusion molecule between the extracellular N-terminal domain of CCR9 and IL-7, which has the potential to overcome these difficulties. This novel fusion protein retains the thymopoietic activity of IL-7 and the ligand-binding ability of CCR9. As a thymopoietic agent, compared with IL-7, it shows an enhanced retention in the thymus, increased de novo T cell production, and increased thymic output. Old mice receiving the fusion protein show improved CD8 T cell responses and reduced viral load after infection with influenza virus compared with those receiving IL-7. This chimeric molecule offers a novel therapeutic strategy that may result in the production of an effective immunorestorative agent. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16148161&query_hl=1 ER - TY - JFULL T1 - Discordance between power Doppler and DAS28 assessment of remission induction in a randomised, placebo-controlled stud of infliximab therapy in early rheumatoid arthritis A1 - Taylor, PC A1 - Steuer, A A1 - McClinton, C A1 - Blomley, M A1 - Wagner, C A1 - Marsters, P A1 - Maini, RN J1 - ARTHRITIS RHEUM Y1 - 2005/09// VL - 52 SN - 0004-3591 SP - S347 EP - S348 ER - TY - JFULL T1 - The distribution of the endogenous retroviruses HERV-K113 and HERV-K115 in health and disease. A1 - Moyes, DL A1 - Martin, A A1 - Sawcer, S A1 - Temperton, N A1 - Worthington, J A1 - Griffiths, DJ A1 - Venables, PJ J1 - Genomics Y1 - 2005/09// VL - 86 SN - 0888-7543 SP - 337 EP - 341 N2 - The human endogenous retroviruses HERV-K113 and HERV-K115 are full-length proviruses but unusual in being found in only a proportion of the population. Here, we study the geographic distribution of these HERVs and their prevalence in autoimmune disease. The frequency of HERV-K113 and HERV-K115 in 174 individuals from Africa was 21.8 and 34.1%, respectively, compared to 4.16 and 1% in 96 people in the United Kingdom (p < 0.001). Prevalence in Yemen (n = 56) was 8 and 7.14% and in Papua New Guinea (n = 54) 0% for both. In the United Kingdom, HERV-K113 was found in 15.6% of 96 Sjögren's syndrome patients (p < 0.01) and 11.9% of 109 multiple sclerosis patients (p < 0.05). No increase in prevalence in either disease was seen with HERV-K115. These data suggest that both viruses are recently integrated and/or under positive evolutionary selection pressure. HERV-K113 may be a genetic risk factor for some types of autoimmunity. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16024218&query_hl=1 ER - TY - JFULL T1 - Infliximab consistently induces clinical remission in patients with early active rheumatoid arthritis regardless of remission criteria. A1 - Smolen, JS A1 - Han, C A1 - Bala, M A1 - van der Heijde, D A1 - Emery, P A1 - Bathon, JM A1 - Keystone, EC A1 - Maini, RN A1 - Kalden, JR A1 - Aletaha, D A1 - Baker, D A1 - Han, J A1 - St Clair, EW J1 - ARTHRITIS RHEUM Y1 - 2005/09// VL - 52 SN - 0004-3591 SP - S139 EP - S139 ER - TY - JFULL T1 - Effect of infliximab and methotrexate on radiographic progression in patients with early rheumatoid arthritis A1 - van der Heijde, D A1 - Emery, P A1 - Keystone, EC A1 - Maini, RN A1 - Durez, P A1 - Kalden, JR A1 - Schiff, M A1 - Han, J A1 - Baker, D A1 - Smolen, JS A1 - St Clair, EW J1 - ARTHRITIS RHEUM Y1 - 2005/09// VL - 52 SN - 0004-3591 SP - S139 EP - S140 ER - TY - JFULL T1 - Patients with early rheumatoid arthritis achieved a clinically meaningful and sustained improvement in physical function after treatment with infliximab. A1 - Smolen, JS A1 - Han, CL A1 - Bala, M A1 - van der Heijde, D A1 - Emery, P A1 - Bathon, JM A1 - Keystone, EC A1 - Maini, RN A1 - Kalden, JR A1 - Baker, D A1 - Han, J A1 - St Clair, EW J1 - ARTHRITIS RHEUM Y1 - 2005/09// VL - 52 SN - 0004-3591 SP - S37 EP - S37 ER - TY - JFULL T1 - Phase I study of TNFalpha AutoVaccIne in patients with metastatic cancer. A1 - Waterston, AM A1 - Gumbrell, L A1 - Bratt, T A1 - Waller, S A1 - Gustav-Aspland, J A1 - L'hermenier, C A1 - Bellenger, K A1 - Campbell, M A1 - Powles, T A1 - Highley, M A1 - Bower, M A1 - Mouritsen, S A1 - Feldmann, M A1 - Coombes, RC J1 - Cancer Immunol Immunother Y1 - 2005/09// VL - 54 SN - 0340-7004 SP - 848 EP - 857 N2 - We evaluated the safety and immunogencity of a novel vaccine directed against autologous TNFalpha in a Phase I fixed dose escalation trial. The vaccine consisted of two recombinant TNFalpha proteins, with specific peptides replaced by foreign immunodominant T cell epitopes from tetanus toxoid. The main objectives were to establish a safe dose and evaluate the vaccines ability to raise neutralising TNFalpha antibodies. Secondary objectives were improvements in body weight and tumour response. Thirty-three patients were vaccinated with three doses (20, 100, or 400 mug) of TNFalpha vaccine at 2-weekly intervals adjuvanted with aluminium hydroxide. Anti-TNFalpha antibody titres were measured by both a RIA, using soluble native TNFalpha as the antigen, and by an ELISA using immobilized partly denatured TNFalpha. Eleven patients (33%) had mild grade1/2 injection site reactions at the higher doses. In 10 of 20 patients, serum antibodies recognize denatured TNFalpha in the ELISA, whereas, antibody titres against native TNFalpha in the RIA were undetectable. This suggests that the production process had partly denatured the vaccine preventing the formation of cross-reacting antibodies to native TNFalpha. In conclusion, TNFalpha vaccine was able to elicit vaccine specific antibodies. However, since the antibodies were only able to cross-react with partly denatured TNFalpha, evaluation of safety and tumour responses to the TNFalpha vaccine was compromised. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15754205&query_hl=1 ER - TY - JFULL T1 - Reduction in radiographic progression in the hands and feet of patients with early rheumatoid arthritis after receiving infliximab in combination with methotrexate A1 - van der Heijde, D A1 - Emery, P A1 - Bathon, JM A1 - Maini, RN A1 - Durez, P A1 - Westhovens, R A1 - Han, J A1 - St Clair, EW A1 - Smolen, JS J1 - ARTHRITIS RHEUM Y1 - 2005/09// VL - 52 SN - 0004-3591 SP - S739 EP - S739 ER - TY - JFULL T1 - Proteomic analysis of T cell-contact dependent activation of macrophages; Identification of an antigen independent immune synapse. A1 - Cope, AP A1 - Peirce, M A1 - Wu, X A1 - Begum, S A1 - Saklatvala, J A1 - Wait, R J1 - ARTHRITIS RHEUM Y1 - 2005/09// VL - 52 SN - 0004-3591 SP - S493 EP - S494 ER - TY - JFULL T1 - The Tec kinases: Bmx and Btk play distinct regulatory roles in osteoclast differentiation A1 - Danks, L A1 - Foxwell, BM A1 - Horwood, NJ J1 - J BONE MINER RES Y1 - 2005/09// VL - 20 SN - 0884-0431 SP - S148 EP - S148 ER - TY - JFULL T1 - Molecular therapeutic targets in rheumatoid arthritis. A1 - Sacre, SM A1 - Andreakos, E A1 - Taylor, P A1 - Feldmann, M A1 - Foxwell, BM J1 - Expert Rev Mol Med Y1 - 2005/08/24/ VL - 7 SN - 1462-3994 SP - 1 EP - 20 N2 - in an attempt to combat the pain and damage generated by rheumatoid arthritis (ra), new drugs are being developed to target molecular aspects of the disease process. recently, a major development has been the use of biologicals (antibodies and soluble receptors) that neutralise the activity of tumour necrosis factor alpha (tnf-alpha) and interleukin 1 (il-1), both of which are involved in disease progression. an increase in our understanding of cell and molecular biology has resulted in the identification and investigation of potential new targets, and also the refinement and improvement of current therapeutic modalities. this review describes therapies that are approved, in clinical trials or under pre-clinical investigation at the laboratory level, particularly focusing on cytokines, although other therapeutic targets of interest are mentioned. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16117838&query_hl=1 ER - TY - JFULL T1 - Palmitoylation at Cys574 is essential for MT1-MMP to promote cell migration. A1 - Anilkumar, N A1 - Uekita, T A1 - Couchman, JR A1 - Nagase, H A1 - Seiki, M A1 - Itoh, Y J1 - FASEB J Y1 - 2005/08// VL - 19 SN - 1530-6860 SP - 1326 EP - 1328 N2 - MT1-MMP is a type I transmembrane proteinase that promotes cell migration and invasion. Here, we report that MT1-MMP is palmitoylated at Cys574 in the cytoplasmic domain, and this lipid modification is critical for its promotion of cell migration and clathrin-mediated internalization. The palmitoylation-defective mutant (C574A) failed to promote cell migration and was not internalized through clathrin pathway like wild-type, but it was internalized through the caveolae pathway. Reintroducing a cysteine at different positions in the cytoplasmic tail of the C574A mutant revealed that the position of the palmitoylated cysteine relative to LLY573, a motif that interacts with mu2 subunit of adaptor protein 2, is critical for the cell motility-promoting activity of MT1-MMP and its clathrin-mediated internalization. Taken together, palmitoylation of MT1-MMP is one of the key posttranslational modifications that determines MT1-MMP-dependent cell migration. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15946988&query_hl=1 ER - TY - JFULL T1 - Synergistic enhancement of Toll-like receptor responses by NOD1 activation. A1 - van Heel, DA A1 - Ghosh, S A1 - Butler, M A1 - Hunt, K A1 - Foxwell, BM A1 - Mengin-Lecreulx, D A1 - Playford, RJ J1 - Eur J Immunol Y1 - 2005/08// VL - 35 SN - 0014-2980 SP - 2471 EP - 2476 N2 - NOD1 is an intracellular pattern-recognition receptor specific for Gram-negative peptidoglycan that is important in host response to infections (e.g. Helicobacter pylori and Shigella flexneri). Genetic variation in NOD1 predisposes to asthma and inflammatory bowel disease. Functional responses have not previously been studied in primary human cells. NOD1 activation by low nanomolar concentrations of the specific muropeptide ligand M-TriDAP induced minimal human peripheral blood mononuclear cell TNF-alpha, IL-1beta or IL-10 secretion, but synergistically increased Toll-like receptor (TLR)-induced responses. Synergistic responses were seen across multiple ligands (to TLR1/2, 2/6, 4, 5, 7/8) and a broad range of cytokine secretion (TNF-alpha, IL-1alpha, IL-1beta, IL-4, IL-6, IL-10, GM-CSF). Synergy was also observed in the allogeneic mixed lymphocyte reaction. These responses were similar in cells homozygous for Crohn's disease-associated NOD2 mutations. In contrast to cell lines, primary human peripheral blood mononuclear cells respond to NOD1 muropeptides at approximately 100-fold lower concentrations. Cross-talk between cytosolic NOD1 and membrane-bound TLR enhances responses to the multiple antigens simultaneously presented by a microbe. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16021603&query_hl=1 ER - TY - JFULL T1 - Competitive disruption of the tumor-promoting function of membrane type 1 matrix metalloproteinase/matrix metalloproteinase-14 in vivo. A1 - Nonaka, T A1 - Nishibashi, K A1 - Itoh, Y A1 - Yana, I A1 - Seiki, M J1 - Mol Cancer Ther Y1 - 2005/08// VL - 4 SN - 1535-7163 SP - 1157 EP - 1166 N2 - Membrane type 1 matrix metalloproteinase (MT1-MMP) is a potent modulator of the pericellular environment and promotes tumor cell invasion and proliferation in many types of tumor. The activation of proMMP-2 and processing of collagen I by MT1-MMP have been thought to be important for its tumor-promoting function. These activities can be inhibited by mutant forms of MT1-MMP lacking the catalytic domain. However, the effect of such dominant-negative mutants has never been evaluated in vivo. Various mutants lacking the catalytic domain (dCAT) were prepared and confirmed to inhibit MT1-MMP activity in human fibrosarcoma HT1080 cells, and tumor cells expressing these mutants were implanted s.c. into nude mice to monitor tumor formation. Only the membrane-anchored form of a dCAT construct through the transmembrane domain [dCAT(1)] showed potent antitumor activity not only in HT1080 cells but also in gastric carcinoma MKN28 and MKN45 cells expressing MT1-MMP. A soluble form of dCAT lacking the transmembrane domain did not show such activity. The expression of dCAT(1) in MKN28 or MKN45 further prevented the metastatic spread of tumor cells into the peritoneal cavity; however, dCAT(1) showed no effect against TMK-1, another gastric carcinoma cell line expressing no MT1-MMP. It is of note that the tumorigenicity of TMK-1 cells enhanced by MT1-MMP overexpression was, in turn, canceled by the additional expression of dCAT(1). Thus, MT1-MMP expressed in tumor cells seems to play a pivotal role in tumor growth in mice. The results also suggest new possibilities to abrogate the tumor-promoting function of MT1-MMP other than the conventional protease inhibitor-based approach. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16093431&query_hl=1 ER - TY - JFULL T1 - Heat shock protein 27 is associated with freedom from graft vasculopathy after human cardiac transplantation. A1 - De Souza, AI A1 - Wait, R A1 - Mitchell, AG A1 - Banner, NR A1 - Dunn, MJ A1 - Rose, ML J1 - Circ Res Y1 - 2005/07/22/ VL - 97 SN - 1524-4571 SP - 192 EP - 198 N2 - Experimental studies have suggested that protective genes protect allografts from cardiac allograft vasculopathy (CAV), the major complication after cardiac transplantation. Here we have sought to confirm this hypothesis using long-term heart transplant recipients. Twenty-two patients that were 9 years or older after transplant were investigated; 11 of these were without angiographic evidence of CAV; 11 had developed early CAV at 1 to 3 years after transplant. To identify proteins that may act as protectors from CAV, a global proteomic approach was used comparing cardiac biopsies from 12 patients taken within the first 2 weeks after transplant and those taken after 9 years from the same patient. Proteins were separated by 2-D gel-electrophoresis, detected by silver staining, and analyzed using Progenesis software. A particular protein spot was found in 4/6 biopsies from patients without CAV, but absent from 5/6 biopsies from those with CAV (P=0.24); however, quantitative analysis of spot intensity showed a significant difference (0.061+/-0.05 versus 0.003+/-0.01, P=0.04). This spot was identified by mass spectrometry and a combination of techniques as a diphosphorylated form of HSP27. Immunohistochemistry of further biopsies not only validated that HSP27 was more abundantly expressed on biopsies without CAV but also showed it to be localized to blood vessels. In contrast, vessels from patients with CAV did not express HSP27 (P=0.028x10(-4)). Immunohistochemistry of 12 further early biopsies and nontransplanted heart showed HSP27 to be present in normal blood vessels. These findings suggest that expression of a specific diphosphorylated form of HSP27 is associated with healthy blood vessels; it appears to be lost from vessels of patients with graft vasculopathy. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15976317&query_hl=1 ER - TY - JFULL T1 - Efficient isolation of peptide ligands for the endothelial cell protein C receptor (EPCR) using candidate receptor phage display biopanning A1 - White, SJ A1 - Simmonds, RE A1 - Lane, DA A1 - Baker, AH J1 - PEPTIDES Y1 - 2005/07// VL - 26 SN - 0196-9781 SP - 1264 EP - 1269 N2 - Phage display biopanning has been used for a number of applications including ligand generation for targeted drug delivery, targeting gene therapy vectors and identification of protein-protein interaction sites. In this study, a random phage display library was used to isolate peptide ligands to the endothelial protein C receptor (EPCR), identifying 74 different peptide sequences and several motifs. Binding to EPCR was characterized by a solid phase binding assay, demonstrating that 95 % of isolated peptides were specific for EPCR. Several homologies with potential relevance to EPCR biology were identified, the most notable being leukolysin (MT-MMP6) and cerastocytin. (c) 2005 Elsevier Inc. All rights reserved. ER - TY - JFULL T1 - Expression of Bruton's tyrosine kinase in osteoclasts A1 - Danks, L A1 - Horwood, NJ J1 - J BONE MINER RES Y1 - 2005/07// VL - 20 SN - 0884-0431 SP - 1309 EP - 1309 ER - TY - JFULL T1 - Effect of infliximab and methotrexate on radiographic progression in patients with early rheumatoid arthritis A1 - Van der Heijde, D A1 - Emery, P A1 - Keystone, E A1 - Maini, RN A1 - Durez, P A1 - Kalden, JR A1 - Schiff, M A1 - Han, J A1 - Baker, D A1 - Smolen, J A1 - St Clair, E J1 - ANN RHEUM DIS Y1 - 2005/07// VL - 64 SN - 0003-4967 SP - 417 EP - 417 ER - TY - JFULL T1 - Comparison of improvement in employability between patients with early RA and patients with established RA after one year of infliximab treatment A1 - Han, C A1 - Bala, M A1 - Smolen, J A1 - StClair, E A1 - Maini, RN A1 - Baker, D J1 - ANN RHEUM DIS Y1 - 2005/07// VL - 64 SN - 0003-4967 SP - 390 EP - 391 ER - TY - JFULL T1 - HB-EGF/HER-1 signaling in bone marrow mesenchymal stem cells: inducing cell expansion and reversibly preventing multilineage differentiation A1 - Krampera, M A1 - Pasini, A A1 - Rigo, A A1 - Scupoli, MT A1 - Tecchio, C A1 - Malpeli, G A1 - Scarpa, A A1 - Dazzi, F A1 - Pizzolo, G A1 - Vinante, F J1 - BLOOD Y1 - 2005/07/01/ VL - 106 SN - 0006-4971 SP - 59 EP - 66 N2 - Epidermal growth factor receptor-1 (EGFR-1/ HER-1/ErbB-1) regulates proliferation and cell fate during epidermal development. HER-1 is activated by several EGF-family ligands including heparin-binding epidermal growth factor-like growth factor (HB-EGF), a mitogenic and chemotactic molecule that participates in tissue repair, tumor growth, and other tissue-modeling phenomena, such as angiogenesis and fibrogenesis. We found that mesenchymal stem cells (MSCS), the precursors of different mesenchymal tissues with a role in processes in which HB-EGF is often involved, normally express HEIR-1, but not HB-EGF itself. Under the effect of HB-EGF, MSCs proliferate more rapidly and persistently, without undergoing spontaneous differentiation. This effect occurs in a dose-dependent fashion, and is specific, direct, and HER-1 mediated, as it is inhibited by anti-HEIR-1 and anti-HB-EGF blocking antibodies. Moreover, HB-EGF reversibly prevents adipogenic, osteogenic, and chondrogenic differentiation induced with specific media. These data show that HB-EGF/HER-1 signaling is relevant to MSC biology, by regulating both proliferation and differentiation. ER - TY - JFULL T1 - Kinetics of change in erosion scores as assessed by plain radiography and high frequency ultrasound in a randomised, controlled trial of infliximab plus MTX versus MTX only therapy in erosive early rheumatoid arthritis A1 - Taylor, PC A1 - Steuer, A A1 - Gruber, J A1 - McClinton, C A1 - Blomley, M A1 - Wagner, C A1 - Marsters, R A1 - Maini, RN J1 - ANN RHEUM DIS Y1 - 2005/07// VL - 64 SN - 0003-4967 SP - 89 EP - 89 ER - TY - JFULL T1 - Reduction in radiographic progression in the hands and feet of patients with early rheumatoid arthritis after receiving infliximab in combination with methotrexate A1 - Van der Heijde, D A1 - Emery, P A1 - Bathon, JM A1 - Maini, RN A1 - Durez, P A1 - Westhovens, R A1 - Han, J A1 - St Clair, E A1 - Smolen, J J1 - ANN RHEUM DIS Y1 - 2005/07// VL - 64 SN - 0003-4967 SP - 418 EP - 419 ER - TY - JFULL T1 - Patients with early rheumatoid arthritis achieved a clinically meaningful and sustained improvement in physical function after treatment with infliximab A1 - Smolen, J A1 - Han, C A1 - Bala, M A1 - Van der Heijde, D A1 - Emery, P A1 - Bathon, JM A1 - Keystone, E A1 - Maini, RN A1 - Kalden, JR A1 - Baker, D A1 - Han, J A1 - St Clair, E J1 - ANN RHEUM DIS Y1 - 2005/07// VL - 64 SN - 0003-4967 SP - 418 EP - 418 ER - TY - JFULL T1 - Steam inhalation treatment for children. A1 - Akhavani, MA A1 - Baker, RH J1 - Br J Gen Pract Y1 - 2005/07// VL - 55 SN - 0960-1643 SP - 557 EP - 557 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16004753&query_hl=1 ER - TY - JFULL T1 - Comparison of remission induction in erosive early KA assessed by DAS28 calculated using either ESR or CRP in infliximab plus MTX combination therapy or MTX monotherapy A1 - Taylor, PC A1 - Steuer, A A1 - McClinton, C A1 - Douglas, J A1 - Wagner, C A1 - Marsters, P A1 - Maini, RN J1 - ANN RHEUM DIS Y1 - 2005/07// VL - 64 SN - 0003-4967 SP - 417 EP - 418 ER - TY - JFULL T1 - Biological therapies and infections: Update and prospects A1 - Maini, RN J1 - ANN RHEUM DIS Y1 - 2005/07// VL - 64 SN - 0003-4967 SP - 3 EP - 3 ER - TY - JFULL T1 - Comparison of protein expression in human deltoideus and vastus lateralis muscles using two-dimensional gel electrophoresis. A1 - Capitanio, D A1 - Viganò, A A1 - Ricci, E A1 - Cerretelli, P A1 - Wait, R A1 - Gelfi, C J1 - Proteomics Y1 - 2005/07// VL - 5 SN - 1615-9853 SP - 2577 EP - 2586 N2 - We have used two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) to study the expression of contractile and regulatory proteins in human vastus lateralis and deltoideus muscles, in order to understand protein turnover and isoform switching in muscles with the same fiber-type composition but different functional properties. We demonstrate a two- to six-fold overexpression of enzymes associated with glycolysis, the tricarboxylic acid cycle, oxidative phosphorylation, and substrate transport in vastus lateralis compared to deltoideus. Expression levels of contractile protein isoforms correlated to the proportion of slow-twitch fibers in deltoideus compared to vastus lateralis are consistent with the different contractile properties of the two muscles. Two proteins involved in free radical homeostasis were differentially expressed, suggesting a direct relationship between radical scavenging and the muscle function. The application of 2-DE and MS to studies of muscle physiology thus offers a more comprehensive assessment of the molecular determinants of muscle function than traditional approaches. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15931664&query_hl=1 ER - TY - JFULL T1 - Inhibition of NF-kappa B and oxidative pathways in human dendritic cells by antioxidative vitamins generates regulatory T cells. A1 - Tan, PH A1 - Sagoo, P A1 - Chan, C A1 - Yates, JB A1 - Campbell, J A1 - Beutelspacher, SC A1 - Foxwell, BM A1 - Lombardi, G A1 - George, AJ J1 - J Immunol Y1 - 2005/06/15/ VL - 174 SN - 0022-1767 SP - 7633 EP - 7644 N2 - Dendritic cells (DCs) are central to T cell immunity, and many strategies have been used to manipulate DCs to modify immune responses. We investigated the effects of antioxidants ascorbate (vitamin C) and alpha-tocopherol (vitamin E) on DC phenotype and function. Vitamins C and E are both antioxidants, and concurrent use results in a nonadditive activity. We have demonstrated that DC treated with these antioxidants are resistant to phenotypic and functional changes following stimulation with proinflammatory cytokines. Following treatment, the levels of intracellular oxygen radical species were reduced, and the protein kinase RNA-regulated, eukaryotic translation initiation factor 2alpha, NF-kappaB, protein kinase C, and p38 MAPK pathways could not be activated following inflammatory agent stimulation. We went on to show that allogeneic T cells (including CD4(+)CD45RO, CD4(+)CD45RA, and CD4(+)CD25(-) subsets) were anergized following exposure to vitamin-treated DCs, and secreted higher levels of Th2 cytokines and IL-10 than cells incubated with control DCs. These anergic T cells act as regulatory T cells in a contact-dependent manner that is not dependent on IL-4, IL-5, IL-10, IL-13, and TGF-beta. These data indicate that vitamin C- and E-treated DC might be useful for the induction of tolerance to allo- or autoantigens. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15944264&query_hl=1 ER - TY - JFULL T1 - Engraftment of allogeneic hematopoietic stem cells requires both inhibition of host-versus-graft responses and 'space' for homeostatic expansion. A1 - Laylor, R A1 - Dewchand, H A1 - Simpson, E A1 - Dazzi, F J1 - Transplantation Y1 - 2005/06/15/ VL - 79 SN - 0041-1337 SP - 1484 EP - 1491 N2 - BACKGROUND: The establishment of host-versus-graft (HvG) tolerance is the primary aim of reduced intensity conditioning (RIC) regimens for allogeneic stem cell transplantation (SCT). It remains to be clarified to what extent recipient myeloablation is fundamental in the establishment of donor chimerism. METHODS: We have addressed this question in a murine model of RIC SCT in which the donor-recipient combination produces HvG against the male specific minor histocompatibility antigen HY. In this system engraftment can be monitored by RT-PCR and HvG effectors enumerated by tetramer analysis. RESULTS: We demonstrate that the dose of irradiation influences donor hemopoietic engraftment and affects generation of anti-donor specific T cells. Chimeric recipients do not mount a HvG immune response, becoming selectively tolerant, as demonstrated by the long term acceptance of skin grafts of donor but not third party origin. However, HvG tolerance is not sufficient to secure engraftment since, even in the absence of HvG, partial myeloablation was still required. The "space" produced by myeloablation and the consequent potential for donor cell expansion could also affect HvG tolerance, since its induction is severely impaired when donor hematopoietic cells have reduced proliferative capacity. CONCLUSIONS: We conclude that both some degree of myeloablation and HvG tolerance are required for successful engraftment, and that the capacity of donor cells to proliferate influences the induction of HvG tolerance. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15940036&query_hl=1 ER - TY - JFULL T1 - Design of effective immunotherapy for human autoimmunity. A1 - Feldmann, M A1 - Steinman, L J1 - Nature Y1 - 2005/06/02/ VL - 435 SN - 1476-4687 SP - 612 EP - 619 N2 - A better understanding of the molecules involved in immune responses has identified many potential targets for the treatment of autoimmune diseases. But although successful therapies have been found for immune disorders in animal studies, few have passed the much harder test of treating human diseases. So far, non-antigen-specific approaches, such as the blocking of tumour-necrosis factor, are achieving some success but the same is not true for antigen-specific approaches. Future therapies will probably include both non-antigen-specific strategies that target cytokines (cell-cell signalling molecules) or block the molecules that stimulate immune responses, and antigen-specific therapies that induce tolerance to self antigens. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15931214&query_hl=1 ER - TY - JFULL T1 - Two-stage affinity purification for inducibly phosphorylated membrane proteins. A1 - Peirce, MJ A1 - Begum, S A1 - Saklatvala, J A1 - Cope, AP A1 - Wait, R J1 - Proteomics Y1 - 2005/06// VL - 5 SN - 1615-9853 SP - 2417 EP - 2421 N2 - Characterisation of tyrosine phosphorylations induced in immune cells in response to inflammatory stimuli may help elucidate the molecular bases of the diversity of immune responses. We have used anti-phosphotyrosine antibodies in combination with cell surface biotinylation in a two-step affinity purification procedure to recover pervanadate-induced tyrosine phosphorylated proteins from sub-cellular compartments, including the cell surface, of murine T cells and macrophages prior to separation by solution-phase isoelectric focussing and one-dimensional gel electrophoresis and identification by tandem mass spectrometry. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15887181&query_hl=1 ER - TY - JFULL T1 - Pathways of T cell activation and terminal differentiation in chronic inflammation. A1 - Isomäki, P A1 - Clark, JM A1 - Panesar, M A1 - Cope, AP J1 - Curr Drug Targets Inflamm Allergy Y1 - 2005/06// VL - 4 SN - 1568-010X SP - 287 EP - 293 N2 - Immune and inflammatory responses are governed by antigen-specific T cells, whose activation, differentiation and effector function are induced by signals delivered via the T cell antigen receptor (TCR) and by costimulatory and cytokine receptors. The molecular events leading to the activation of naïve T cells have been extensively studied and are well characterized. Much less is known about the molecular and biochemical events regulating the activation of T cells in chronic inflammatory diseases such as rheumatoid arthritis (RA). This review examines the current state of knowledge of T cell activation in chronic inflammation, focusing on RA, and summarizes experimental data which indicate that the chronic inflammatory process may profoundly affect TCR and cytokine signal transduction pathways. We present evidence suggesting that in chronic inflammation, the antigen-driven TCR-mediated processes are attenuated, while cytokine-driven effector responses are sustained or even enhanced. The possible implications of this inbalance are discussed. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16101535&query_hl=1 ER - TY - JFULL T1 - Analysis of Lupinus albus storage proteins by two-dimensional electrophoresis and mass spectrometry. A1 - Wait, R A1 - Gianazza, E A1 - Brambilla, D A1 - Eberini, I A1 - Morandi, S A1 - Arnoldi, A A1 - Sirtori, CR J1 - J Agric Food Chem Y1 - 2005/06/01/ VL - 53 SN - 0021-8561 SP - 4599 EP - 4606 N2 - A laboratory-prepared total protein extract (TPE) and a lupin protein isolate (LPI-E) produced in a pilot plant were submitted to a detailed two-dimensional (2DE) proteomic investigation. Recent findings have indicated that in an established rodent model of hyperlipidemia, moderate daily intakes of LPI-Es lead to a reduction of total and low-density lipoprotein cholesterol levels, and the knowledge of the actual composition of the protein sample used in that study is at the basis of further structure/action investigations. The experimental results indicate that the semi-industrial procedure used for the production of LPI-E damages only marginally the proteins. It does, however, cleave some disulfide bridges and induce mild proteolysis, as confirmed by the higher number of resolved protein spots in the low Mr and acidic pI region of the 2DE map. Out of 72 spots submitted to mass spectrometry and compared with available protein databases, 42 correspond to fragments of beta-conglutin, the 7S globulin of lupin, spanning between positions 37 and 495 of the protein sequence. Using the bioinformatic tool BlastP, these peptides were compared to the alpha'-subunit of beta-conglycinin, the 7S globulin of soybean, this being the most active hypocholesterolemic component of soybean protein, as shown by in vitro and in vivo experiments. At least 18 peptides derived from beta-conglutin, having a percentage identity higher than 50% and a similarity percentage higher than 70% vs the alpha'-subunit of beta-conglycinin, are likely candidates to be the biologically active components of lupin protein. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15913332&query_hl=1 ER - TY - JFULL T1 - Serum vascular markers and vascular imaging in assessment of rheumatoid arthritis disease activity and response to therapy. A1 - Taylor, PC J1 - Rheumatology (Oxford) Y1 - 2005/06// VL - 44 SN - 1462-0324 SP - 721 EP - 728 N2 - Vascular pathology, in the form of angiogenesis, is important in the perpetuation of rheumatoid arthritis (RA) and, in the form of endothelial dysfunction, contributes to associated cardiovascular co-morbidity. Emerging evidence suggests that TNFalpha blockade may modify vascular pathology in RA. Serum concentrations of vascular endothelial growth factor (VEGF), a potent endothelial cell-specific growth factor that is up-regulated by pro-inflammatory cytokines and by hypoxia, are elevated in RA and correlate with disease activity. Serum levels of VEGF at first presentation in RA predict radiographic progression of the disease over the subsequent year. Power Doppler ultrasonography is a sensitive method for demonstrating the presence of blood flow in small vessels and the vascular signal correlates with histopathological quantification of the vascular density of synovial tissue. Recent data indicate that high-frequency ultrasound and power Doppler are sensitive tools for evaluation of disease activity and assessment of response to therapy. Power Doppler imaging may also have the potential to predict those patients most at risk of accelerated joint destruction. However, much work has yet to be done to standardize the use of these imaging technologies. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15644394&query_hl=1 ER - TY - JFULL T1 - Intestinal epithelial barrier and mucosal immunity A1 - Didierlaurent, A A1 - Simonet, M A1 - Sirard, JC J1 - CMLS-CELL MOL LIFE S Y1 - 2005/06// VL - 62 SN - 1420-682X SP - 1285 EP - 1287 ER - TY - JFULL T1 - Proteomic analysis of cell surface proteins from Clostridium difficile. A1 - Wright, A A1 - Wait, R A1 - Begum, S A1 - Crossett, B A1 - Nagy, J A1 - Brown, K A1 - Fairweather, N J1 - Proteomics Y1 - 2005/06// VL - 5 SN - 1615-9853 SP - 2443 EP - 2452 N2 - Clostridium difficile is a bacterium that causes disease of the large intestine, particularly after treatment with antibiotics. The bacterium produces two toxins (A and B) that are responsible for the pathology of the disease. In addition, a number of bacterial virulence factors associated with adhesion to the gut have previously been identified, including the cell wall protein Cwp66, the high-molecular weight surface layer protein (HMW-SLP) and the flagella. As the genome sequence predicts many other cell wall associated proteins, we have investigated the diversity of proteins in cell wall extracts, with the aim of identifying further virulence factors. We have used a number of methods to remove the proteins associated with the cell wall of C. difficile. Two of the resulting extracts, obtained using low pH glycine treatment and lysozyme digestion of the cell wall, have been analysed in detail by two-dimensional electrophoresis and mass spectrometry. One hundred and nineteen spots, comprising 49 different proteins, have been identified. The two surface layer proteins (SLPs) are the most abundant proteins, and we have also found components of the flagellum. Interestingly, we have also determined that a number of paralogs of the HMW-SLP are expressed, and these could represent targets for further investigation as virulence factors. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15887182&query_hl=1 ER - TY - JFULL T1 - Effects of prednisolone treatment on cytokine expression in patients with leprosy type 1 reactions. A1 - Andersson, AK A1 - Chaduvula, M A1 - Atkinson, SE A1 - Khanolkar-Young, S A1 - Jain, S A1 - Suneetha, L A1 - Suneetha, S A1 - Lockwood, DN J1 - Infect Immun Y1 - 2005/06// VL - 73 SN - 0019-9567 SP - 3725 EP - 3733 N2 - Leprosy type 1 reactions (T1R) are due to increased cell-mediated immunity and result in localized tissue damage. The anti-inflammatory drug prednisolone is used for treatment, but there is little good in vivo data on the molecular actions of prednisolone. We investigated the effect of prednisolone treatment on tumor necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta), IL-10, and transforming growth factor beta1 (TGF-beta1) mRNA and protein expression in blood and skin biopsies from 30 patients with T1R in India. After 1 month of prednisolone treatment the sizes of the skin granulomas were reduced, as were the grades of cells positive for TNF-alpha and IL-10 in skin lesions. Increased production of TGF-beta1 was seen in skin lesions after 6 months of prednisolone treatment. Expression of mRNA for TNF-alpha, IL-1beta, and TGF-beta1 was reduced, whereas no change in IL-10 mRNA expression was detected during treatment. The circulating cytokine profiles were similar in patients with and without T1R, and prednisolone treatment had no detectable effects on cytokine expression in the blood. The data emphasize the compartmentalization of pathology in T1R and the importance of the immune response in the skin. Clinical improvement and cytokine expression were compared. Surprisingly, patients with improved skin and nerve function and patients with nonimproved skin and nerve function had similar cytokine profiles, suggesting that clinical improvement is not directly mediated by the cytokines studied here. This in vivo well-controlled study of the immunosuppressive effects of prednisolone showed that the drug does not switch off cytokine responses effectively. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15908402&query_hl=1 ER - TY - JFULL T1 - Geographic information system localization of community-acquired MRSA soft tissue abscesses. A1 - Tirabassi, MV A1 - Wadie, G A1 - Moriarty, KP A1 - Garb, J A1 - Konefal, SH A1 - Courtney, RA A1 - Sachs, BF A1 - Wait, R J1 - J Pediatr Surg Y1 - 2005/06// VL - 40 SN - 1531-5037 N2 - BACKGROUND: Soft tissue infections with methicillin-resistant Staphylococcus aureus (MRSA) pose an ever-increasing risk to children in the community. Although historically these infections were limited to children with prolonged hospitalization, the authors have seen an increase in community-acquired infections in children without identifiable risk factors. The goal of this study is to determine the incidence of truly community-acquired MRSA soft tissue infections in our community and geographically map regions of increased risk. METHODS: After obtaining the institutional review board's approval, a retrospective chart review was conducted on 195 patients records who underwent an incision and drainage of soft tissue infections from January 1, 2000, to December 31, 2003. Thirteen patients were excluded from the study because no cultures were taken at the time of incision and drainage. RESULTS: The most common organism isolated from wound culture was S aureus , 40% (73/182), of which 45% (33/73) were MRSA. Eighty-one percent (27/33) of MRSA infections were in Springfield, 1 of 18 towns represented in the patient population. Geographic information system analysis identified a significant MRSA cluster 1.96 km in diameter within the city of Springfield. CONCLUSIONS: Geography proved to be a significant risk factor for presenting with MRSA infection. Geographic maps of antibiotic resistance can be used to guide physician antibiotic selection before culture results are available. This has significant implications for the health care provider in proper antibiotic selection within the community. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15991178&query_hl=1 ER - TY - JFULL T1 - Matrix metalloproteinase expression and function during fin regeneration in zebrafish: analysis of MT1-MMP, MMP2 and TIMP2. A1 - Bai, S A1 - Thummel, R A1 - Godwin, AR A1 - Nagase, H A1 - Itoh, Y A1 - Li, L A1 - Evans, R A1 - McDermott, J A1 - Seiki, M A1 - Sarras, MP J1 - Matrix Biol Y1 - 2005/06// VL - 24 SN - 0945-053X SP - 247 EP - 260 N2 - Matrix metalloproteinases (MMPs) play key roles in the turnover of extracellular matrix (ECM) and, thereby, function as key regulators of cell-ECM interactions during development. In spite of their importance during developmental processes, relatively little has been reported about the role of these metalloproteinases during limb development and regeneration. To approach the problem of cell-ECM interactions during limb (fin) regeneration, we have utilized zebrafish as an experimental model. Based on previous MMP cloning studies from our laboratory, the current study has focused on the expression of membrane-type 1 metalloproteinase (MT1-MMP), gelatinase A (MMP-2) and endogenous tissue inhibitor 2 of metalloproteinases (TIMP-2) during fin regeneration in adult zebrafish. In situ analysis indicated co-expression of zmt1-mmp, zmmp-2, and ztimp-2 mRNA transcripts in regenerating caudal fins. In situ gelatin-zymography confirmed the presence of active metalloproteinases in regenerating fins. zmt1-mmp, zmmp-2, and ztimp-2 mRNA transcripts were expressed in the blastema and basal epithelium during caudal fin regeneration while expression of type IV collagen [zcol-IV(a5)] transcripts (a basal lamina component) was restricted to the basal epithelium. Fin outgrowth was greatly reduced in the presence of GM6001 (an inhibitor of MMP activity) indicating the importance of these enzymes during fin regeneration. Previous studies by Itoh (EMBO, 2001) indicated that expression of a vertebrate MT1-MMP construct containing only the hemopexin-transmembrane-cytoplasmic domains (MT1HPX) resulted in blockage of MT1-MMP homophilic complex formation and subsequent inhibition of pro-MMP-2 activation. Interference with homophilic complex formation was attributed to expression of the hemopexin domain at the cell surface. Building upon these earlier findings, the current study found that ectopic expression of MT1HPX in fin regenerates inhibited the regeneration process and resulted in a reduction in cell proliferation in the blastema. Taken together, these results indicate that MMPs have an important role during fin regeneration in zebrafish. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15935631&query_hl=1 ER - TY - JFULL T1 - Muramyl dipeptide and toll-like receptor sensitivity in NOD2-associated Crohn's disease. A1 - van Heel, DA A1 - Ghosh, S A1 - Butler, M A1 - Hunt, KA A1 - Lundberg, AM A1 - Ahmad, T A1 - McGovern, DP A1 - Onnie, C A1 - Negoro, K A1 - Goldthorpe, S A1 - Foxwell, BM A1 - Mathew, CG A1 - Forbes, A A1 - Jewell, DP A1 - Playford, RJ J1 - Lancet Y1 - 2005/05/21/ VL - 365 SN - 1474-547X SP - 1794 EP - 1796 N2 - Both NOD2 (CARD15) alleles are mutated in roughly 15% of patients with Crohn's disease, but functional effects are unclear. We analysed the cytokine response of peripheral blood mononuclear cells to muramyl dipeptide (MDP), the ligand for NOD2. MDP induced little TNFalpha or interleukin 1beta, but strong interleukin-8 secretion. MDP also substantially upregulated secretion of TNFalpha and interleukin 1beta induced by toll-like receptor ligands. These effects were abolished by the most common Crohn's NOD2 double mutant genotypes at low nanomolar MDP concentrations, and provide the basis to develop a test of NOD2 functional deficiency. In Crohn's disease, there are defects in neutrophil recruitment driven by NOD2 and interleukin 8 and in cross talk between the NOD2 and toll-like receptor pathways, which suggests that the immune system fails to receive an early priming signal. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15910952&query_hl=1 ER - TY - JFULL T1 - Hypoxia and angiogenesis in rheumatoid arthritis. A1 - Taylor, PC A1 - Sivakumar, B J1 - Curr Opin Rheumatol Y1 - 2005/05// VL - 17 SN - 1040-8711 SP - 293 EP - 298 N2 - PURPOSE OF REVIEW: Angiogenesis is a prominent feature of rheumatoid synovitis. Although new blood vessels deliver oxygen to the augmented inflammatory cell mass, the neovascular network is dysfunctional and fails to restore tissue oxygen homeostasis, so that the rheumatoid joint remains a markedly hypoxic environment. The purpose of this review is to discuss the role of hypoxia and angiogenesis in the pathogenesis of rheumatoid arthritis. RECENT FINDINGS: Vascular pathologic change, in the form of angiogenesis, is important in the perpetuation of rheumatoid arthritis and, in the form of endothelial dysfunction, contributes to associated cardiovascular comorbidity. Recent data suggest that tumor necrosis factor-alpha blockade may modify the vascular pathologic changes in rheumatoid arthritis. Angiogenesis is a prominent feature of rheumatoid synovitis. Emerging evidence based on ultrasonographic vascular imaging and angiogenic biomarkers implicates angiogenesis in the active phase of erosive disease. Many factors contribute to the profoundly hypoxic environment that can arise within the joint affected by rheumatoid arthritis. At a cellular level, hypoxia is detected by a mechanism that regulates cytoplasmic concentrations of hypoxia-inducible factor-1alpha. After translocation to the nucleus, hypoxia-inducible factor-1alpha binds its partner hypoxia-inducible factor-1beta to form a heterodimeric, functional transcription factor, hypoxia-inducible factor-1, which activates a gene program associated with angiogenesis, glycolysis, and adaptation to pH. SUMMARY: Despite the luxuriant vasculature associated with rheumatoid arthritis synovitis, the joint affected by rheumatoid arthritis is hypoxic. Repetitive cycles of hypoxia and reoxygenation together with oxidants produced by phagocytic cells promote chronic oxidative stress within the microenvironment of the affected joint, leading to the generation of reactive oxygen species with the potential to contribute to tissue damage. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15838239&query_hl=1 ER - TY - JFULL T1 - The indirect pathway of allorecognition is actively regulated by CD4+CD25+ regulatory T cells. A1 - Notley, CA A1 - Robinson, DS A1 - Dallman, MJ J1 - AM J TRANSPLANT Y1 - 2005/05// VL - 5 SN - 1600-6135 SP - 498 EP - 499 ER - TY - JFULL T1 - Redox options in two-dimensional electrophoresis. A1 - Wait, R A1 - Begum, S A1 - Brambilla, D A1 - Carabelli, AM A1 - Conserva, F A1 - Rocco Guerini, A A1 - Eberini, I A1 - Ballerio, R A1 - Gemeiner, M A1 - Miller, I A1 - Gianazza, E J1 - Amino Acids Y1 - 2005/05// VL - 28 SN - 0939-4451 SP - 239 EP - 272 N2 - Two-dimensional electrophoresis is usually run on fully reduced samples. Under these conditions even covalently bound oligomers are dissociated and individual polypeptide chains may be fully unfolded by both, urea and SDS, which maximizes the number of resolved components and allows their pI and M(r) to be most accurately evaluated. However, various electrophoretic protocols for protein structure investigation require a combination of steps under varying redox conditions. We review here some of the applications of these procedures. We also present some original data about a few related samples -- serum from four species: Homo sapiens, Mus musculus, Rattus norvegicus, Bos taurus -- which we run under fully unreduced and fully reduced conditions as well as with reduction between first and second dimension. We demonstrate that in many cases the unreduced proteins migrate with a better resolution than reduced proteins, mostly in the crowded 'alpha-globulin' area of pI 4.5-6 and M(r) 50-70 kDa. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15744479&query_hl=1 ER - TY - JFULL T1 - The 3' untranslated region of tumor necrosis factor alpha mRNA is a target of the mRNA-stabilizing factor HuR (vol 21, pg 721, 2001) A1 - Dean, JLE A1 - Wait, R A1 - Mahtani, KR A1 - Sully, G A1 - Clark, AR A1 - Saklatvala, J J1 - MOL CELL BIOL Y1 - 2005/04// VL - 25 SN - 0270-7306 SP - 3400 EP - 3400 ER - TY - JFULL T1 - Bone marrow mesenchymal stem cells induce division arrest anergy of activated T cells. A1 - Glennie, S A1 - Soeiro, I A1 - Dyson, PJ A1 - Lam, EW A1 - Dazzi, F J1 - Blood Y1 - 2005/04/01/ VL - 105 SN - 0006-4971 SP - 2821 EP - 2827 N2 - It has been shown that mesenchymal stem cells (MSCs) induce T cells to become unresponsive. We characterized the phenotype of these T cells by dissecting the effect of MSCs on T-cell activation, proliferation, and effector function. For this purpose, an in vitro murine model was used in which T-cell responses were generated against the male HY minor histocompatibility antigen. In the presence of MSCs, the expression of early activation markers CD25 and CD69 was unaffected but interferon-gamma (IFN-gamma) production was reduced. The inhibitory effect of MSCs was directed mainly at the level of cell proliferation. Analysis of the cell cycle showed that T cells, stimulated in the presence of MSCs, were arrested at the G1 phase. At the molecular level, cyclin D2 expression was profoundly inhibited, whereas p27(kip1) was up-regulated. When MSCs were removed from the cultures and restimulated with the cognate peptide, T cells produced IFN-gamma but failed to proliferate. The addition of exogenous interleukin-2 (IL-2) did not restore proliferation. MSCs did not preferentially target any T-cell subset, and the inhibition was also extended to B cells. MSC-mediated inhibition induces an unresponsive T-cell profile that is fully consistent with that observed in division arrest anergy. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15591115&query_hl=1 ER - TY - JFULL T1 - Low level BCR-ABL transcript positivity in long term survivors of allogeneic stem cell transplantation for CIVIL A1 - Patel, KC A1 - Kaeda, J A1 - Dazzi, F A1 - O'Shea, D A1 - Goldman, J A1 - Kanfer, E A1 - Marin, D A1 - Olavarria, E A1 - Rahemtulla, A A1 - Apperley, J A1 - Salooja, N J1 - BRIT J HAEMATOL Y1 - 2005/04// VL - 129 SN - 0007-1048 SP - 75 EP - 75 ER - TY - JFULL T1 - Evidence of radiographic benefit of treatment with infliximab plus methotrexate in rheumatoid arthritis patients who had no clinical improvement: a detailed subanalysis of data from the anti-tumor necrosis factor trial in rheumatoid arthritis with concomitant therapy study. A1 - Smolen, JS A1 - Han, C A1 - Bala, M A1 - Maini, RN A1 - Kalden, JR A1 - van der Heijde, D A1 - Breedveld, FC A1 - Furst, DE A1 - Lipsky, PE A1 - ATTRACT Study Group J1 - Arthritis Rheum Y1 - 2005/04// VL - 52 SN - 0004-3591 SP - 1020 EP - 1030 N2 - OBJECTIVE: To assess the relationship between inflammation and joint destruction in rheumatoid arthritis (RA) patients who have not responded clinically to treatment. METHODS: Changes from baseline to week 54 in clinical variables and measures of radiographic progression were compared between patients who received infliximab (3 mg/kg or 10 mg/kg every 4 or 8 weeks) plus methotrexate (MTX) and those who received MTX plus placebo in the Anti-Tumor Necrosis Factor Trial in RA with Concomitant Therapy trial. RESULTS: At week 54, patients who did not show 20% improvement by American College of Rheumatology criteria (ACR20 nonresponders) while receiving infliximab plus MTX exhibited mild but statistically significant improvement in clinical variables, including the 28-joint Disease Activity Score (DAS28) (P < 0.001), tender joint count (P = 0.014), swollen joint count (P < 0.001), and C-reactive protein (CRP) level (P < 0.001). Whereas the clinical and CRP changes among ACR20 nonresponders to infliximab plus MTX were small and much lower than among ACR20 responders to this treatment, radiographic progression among ACR20 nonresponders to infliximab plus MTX was significantly inhibited (P < 0.001) compared with ACR20 nonresponders to MTX plus placebo. Radiographic progression was much greater in patients receiving MTX plus placebo than in patients receiving infliximab plus MTX, irrespective of ACR response status (mean change in modified Sharp/van der Heijde score 6.0 in ACR20 responders and 7.2 in ACR20 nonresponders in the MTX plus placebo-treated group, versus 0.1 in ACR20 responders and 1.2 in ACR20 nonresponders in the infliximab plus MTX-treated group). Furthermore, among patients who were ACR20 nonresponders through week 54, patients who were DAS nonresponders at weeks 30 and 54, and patients without any improvement in individual clinical variables, those receiving infliximab plus MTX still demonstrated inhibition of structural damage that was statistically significant compared with inhibition in patients who received MTX plus placebo (P < 0.05 to P < 0.001). CONCLUSION: Even in patients without clinical improvement, treatment with infliximab plus MTX provided significant benefit with regard to the destructive process, suggesting that in such patients these 2 measures of disease are dissociated. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15818697&query_hl=1 ER - TY - JFULL T1 - Monitoring patients in complete cytogenetic remission after treatment of CML in chronic phase with imatinib: patterns of residual leukaemia and prognostic factors for cytogenetic relapse. A1 - Marin, D A1 - Kaeda, J A1 - Szydlo, R A1 - Saunders, S A1 - Fleming, A A1 - Howard, J A1 - Andreasson, C A1 - Bua, M A1 - Olavarria, E A1 - Rahemtulla, A A1 - Dazzi, F A1 - Kanfer, E A1 - Goldman, JM A1 - Apperley, JF J1 - Leukemia Y1 - 2005/04// VL - 19 SN - 0887-6924 SP - 507 EP - 512 N2 - We monitored BCR-ABL transcript levels by quantitative real-time PCR in 103 patients treated with imatinib for chronic myeloid leukaemia in chronic phase for a median of 30.3 months (range 5.5-49.9) after they achieved complete cytogenetic remission (CCyR). The patients could be divided into three groups: (1) in 32 patients transcript levels continued to decline during the period of observation (nadir BCR-ABL/ABL ratio 0.015%); in five of these patients BCR-ABL transcripts became undetectable on repeated testing, (2) in 42 patients the transcript levels reached a plateau and (3) in 26 patients transcript numbers increased and the initial CCyR was lost. Three patients were not evaluable. Patients who remained in CCyR for at least 24 months appeared to have a low risk of subsequent cytogenetic relapse. We conclude that the pattern of 'residual' disease after achieving CCyR on imatinib is variable: some patients in CCyR show a progressive reduction in the level of residual disease, some reach a plateau where transcript numbers are relatively stable and others relapse with Ph-positive metaphases. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15703781&query_hl=1 ER - TY - JFULL T1 - Protection against tetanus toxin using a plant-based vaccine. A1 - Tregoning, JS A1 - Clare, S A1 - Bowe, F A1 - Edwards, L A1 - Fairweather, N A1 - Qazi, O A1 - Nixon, PJ A1 - Maliga, P A1 - Dougan, G A1 - Hussell, T J1 - Eur J Immunol Y1 - 2005/04// VL - 35 SN - 0014-2980 SP - 1320 EP - 1326 N2 - Plant-expressed vaccines may provide a unique opportunity for generating anti-pathogen immunity, especially in countries where cold storage is lacking. In the following study, we show that soluble protein from tobacco leaves expressing fragment C of tetanus toxin protected mice against a lethal tetanus toxin challenge. More importantly, we show that a single intranasal (i.n.) vaccination was as efficient as oral delivery, inducing high levels of activated CD4(+) T cells and anti-toxin antibody. Unlike the oral route, i.n. delivery did not require the presence of adjuvant (cholera toxin). Indeed, addition of cholera toxin induced bystander immune responses to plant proteins as well. This is the first study documenting protective immunity by a single i.n. dose of plant vaccine. Plant-based vaccines are promising because they are more heat stable, are easy to produce, cheap and do not require needles. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15739166&query_hl=1 ER - TY - JFULL T1 - Proteome analysis of the anterior cingulate cortex in major psychiatric disorders A1 - Cotter, DR A1 - Beasley, CL A1 - Pennington, KP A1 - Wait, R A1 - Dunn, MJ J1 - SCHIZOPHRENIA BULL Y1 - 2005/04// VL - 31 SN - 0586-7614 SP - 249 EP - 250 ER - TY - JFULL T1 - Fusing subunit antigens to interleukin-2 and encapsulating them in liposomes improves their antigenicity but not their protective efficacy. A1 - Wales, JR A1 - Baird, MA A1 - Davies, NM A1 - Buchan, GS J1 - Vaccine Y1 - 2005/03/18/ VL - 23 SN - 0264-410X SP - 2339 EP - 2341 N2 - Subunit vaccines commonly lack sufficient immunogenicity to stimulate a comprehensive protective immune response in vivo. We have investigated the potential of specific cytokines (interleukin-2) and particulate delivery systems (liposomes) to enhance antigenicity. Here we report that the IgG1 and IFN-gamma responses to a subunit antigen, consisting of a T and B-cell epitope from Influenza haemagglutinin, can be improved when it is both fused to interelukin-2 and encapsulated in liposomes. However, this vaccine formulation was not able to protect animals against a challenge with live Influenza A/PR/8/34 virus. The addition of more potent immune stimulators may be necessary to improve responses. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15755624&query_hl=1 ER - TY - JFULL T1 - Dynamic kinetic resolution of racemates using membrane-separated catalysts. A1 - Gibbins, E A1 - Irwin, JL A1 - Livingston, AG A1 - Muir, JC A1 - Patterson, DA A1 - Roengpithya, C A1 - Taylor, PC J1 - ABSTR PAP AM CHEM S Y1 - 2005/03/13/ VL - 229 SP - U550 EP - U550 ER - TY - JFULL T1 - X-ray structure of human proMMP-1: new insights into procollagenase activation and collagen binding. A1 - Jozic, D A1 - Bourenkov, G A1 - Lim, NH A1 - Visse, R A1 - Nagase, H A1 - Bode, W A1 - Maskos, K J1 - J Biol Chem Y1 - 2005/03/11/ VL - 280 SN - 0021-9258 SP - 9578 EP - 9585 N2 - Vertebrate collagenases, members of the matrix metalloproteinase (MMP) family, initiate interstitial fibrillar collagen breakdown. It is essential in many biological processes, and unbalanced collagenolysis is associated with diseases such as arthritis, cancer, atherosclerosis, aneurysm, and fibrosis. These metalloproteinases are secreted from the cell as inactive precursors, procollagenases (proMMPs). To gain insights into the structural basis of their activation mechanisms and collagen binding, we have crystallized recombinant human proMMP-1 and determined its structure to 2.2 A resolution. The catalytic metalloproteinase domain and the C-terminal hemopexin (Hpx) domain show the classical MMP-fold, but the structure has revealed new features in surface loops and domain interaction. The prodomain is formed by a three-helix bundle and gives insight into the stepwise activation mechanism of proMMP-1. The prodomain interacts with the Hpx domain, which affects the position of the Hpx domain relative to the catalytic domain. This interaction results in a "closed" configuration of proMMP-1 in contrast to the "open" configuration observed previously for the structure of active MMP-1. This is the first evidence of mobility of the Hpx domain in relation to the catalytic domain, providing an important clue toward the understanding of the collagenase-collagen interaction and subsequent collagenolysis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15611040&query_hl=1 ER - TY - JFULL T1 - Spontaneous chemical reversion of an active site mutation: deamidation of an asparagine residue replacing the catalytic aspartic acid of glutamate dehydrogenase. A1 - Paradisi, F A1 - Dean, JL A1 - Geoghegan, KF A1 - Engel, PC J1 - Biochemistry Y1 - 2005/03/08/ VL - 44 SN - 0006-2960 SP - 3636 EP - 3643 N2 - A mutant (D165N) of clostridial glutamate dehydrogenase (GDH) in which the catalytic Asp is replaced by Asn surprisingly showed a residual 2% of wild-type activity when purified after expression in Escherichia coli at 37 degrees C. This low-level activity also displayed Michaelis constants for substrates that were remarkably similar to those of the wild-type enzyme. Expression at 8 degrees C gave a mutant enzyme preparation 1000 times less active than the first preparation, but progressively, over 2 weeks' incubation at 37 degrees C in sealed vials, this enzyme regained 90% of the specific activity of wild type. This suggested that the mutant might undergo spontaneous deamidation. Mass spectrometric analysis of tryptic peptides derived from D165N samples treated in various ways showed (i) that the Asn is in place in D165N GDH freshly prepared at 8 degrees C; (ii) that there is a time-dependent reversion of this Asn to Asp over the 2-week incubation period; (iii) that detectable deamidation of other Asn residues, in Asn-Gly sequences, mainly occurred in sample workup rather than during the 2-week incubation; (iv) that there is no significant deamidation of other randomly chosen Asn residues in this mutant over the same period; and (v) that when the protein is denatured before incubation, no deamidation at Asn-165 is detectable. It appears that this deamidation depends on the residual catalytic machinery of the mutated GDH active site. A literature search indicates that this finding is not unique and that Asn may not be a suitable mutational replacement in the assessment of putative catalytic Asp residues by site-directed mutagenesis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15736973&query_hl=1 ER - TY - JFULL T1 - Muscle protein expression changes during extremely high altitude exposure A1 - Cerretelli, P A1 - Ripamonti, M A1 - De Palma, S A1 - Wait, R A1 - Howald, H A1 - Hoppeler, H A1 - Gelfi, C J1 - FASEB J Y1 - 2005/03/04/ VL - 19 SN - 0892-6638 SP - A745 EP - A745 ER - TY - JFULL T1 - Molecular determinants of muscle plasticity: Human single fiber proteornic differential analysis A1 - Gelfi, C A1 - Vigano, A A1 - Ripamonti, M A1 - Capitanio, D A1 - Wait, R A1 - Bottinelli, R A1 - Cerretelli, P J1 - FASEB J Y1 - 2005/03/04/ VL - 19 SN - 0892-6638 SP - A117 EP - A118 ER - TY - JFULL T1 - Does post-transplant treatment with imatinib mesylate inhibit graft-versus-leukemia? Reply A1 - Cwynarski, K A1 - Melo, JV A1 - Dazzi, F J1 - LEUKEMIA Y1 - 2005/03// VL - 19 SN - 0887-6924 SP - 457 EP - 457 ER - TY - JFULL T1 - Irradiation influences endothelial cell function in vitro and in vivo: a possible role of IP10 A1 - Cannella, L A1 - Laylor, R A1 - Marelli-Berg, F A1 - Dazzi, F J1 - BONE MARROW TRANSPL Y1 - 2005/03// VL - 35 SN - 0268-3369 SP - S342 EP - S342 ER - TY - JFULL T1 - TGF-beta prevents eosinophilic lung disease but impairs pathogen clearance. A1 - Williams, AE A1 - Humphreys, IR A1 - Cornere, M A1 - Edwards, L A1 - Rae, A A1 - Hussell, T J1 - Microbes Infect Y1 - 2005/03// VL - 7 SN - 1286-4579 SP - 365 EP - 374 N2 - Respiratory infections are the third leading cause of death worldwide. Complications arise directly as a consequence of pathogen replication or indirectly due to aberrant or excessive immune responses. In the following report, we evaluate the efficacy, in a murine model, of nasally delivered DNA encoding TGF-beta1 to suppress immunopathology in response to a variety of infectious agents. A single nasal administration suppressed lymphocyte responses to Cryptococcus neoformans, influenza virus and respiratory syncytial virus. The suppression did not depend on the phenotype of the responding T cell, since both Th1 and Th2 responses were affected. During Th2-inducing infection, pulmonary eosinophilic responses were significantly suppressed. In all cases, however, suppressed immunity correlated with increased susceptibility to infection. We conclude that nasal TGF-beta treatment could be used to prevent pulmonary, pathogen-driven eosinophilic disease, although anti-pathogen strategies will need to be administered concordantly. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15784186&query_hl=1 ER - TY - JFULL T1 - NK cells support osteoclast formation in vitro A1 - Danks, L A1 - Owen, S A1 - Feldmann, M A1 - Brennan, F A1 - Horwood, NJ J1 - RHEUMATOLOGY Y1 - 2005/03// VL - 44 SN - 1462-0324 SP - I30 EP - I30 ER - TY - JFULL T1 - Immunotherapy of rheumatoid arthritis: past, present and future. A1 - Sivakumar, B A1 - Paleolog, E J1 - Curr Opin Drug Discov Devel Y1 - 2005/03// VL - 8 SN - 1367-6733 SP - 169 EP - 176 N2 - Rheumatoid arthritis (RA) is a chronic autoimmune disease, characterized by inflammation of the synovial lining of joints, and the destruction of cartilage and bone. Seminal studies demonstrating that pro-inflammatory cytokines, such as tumor necrosis factor-alpha (TNFalpha), are expressed in RA, has resulted in the approval of anti-TNFalpha biological therapies for its treatment. Although groundbreaking in themselves, these studies have also paved the way for further research to determine whether the targeting of other cytokines and immune pathways might aid in development of the next generation of drugs for the treatment of RA. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15782541&query_hl=1 ER - TY - JFULL T1 - Ceramide glycosylation and fatty acid hydroxylation influence serological reactivity in Trypanosoma cruzi glycosphingolipids. A1 - Villas-Boas, MH A1 - Wait, R A1 - Silva, RB A1 - Rodrigues, ML A1 - Barreto-Bergter, E J1 - FEMS Microbiol Lett Y1 - 2005/03/01/ VL - 244 SN - 0378-1097 SP - 47 EP - 52 N2 - Ceramide mono (CMH) or dihexoside (CDH) fractions from Trypanosoma cruzi (Dm28c clone) were identified as glucosyl and lactosylceramides containing non-hydroxylated fatty acids. The di-glycosylated form was much more efficiently recognized by sera from T. cruzi-immunized rabbits, indicating that glycosylation influences antigenicity. Fatty acid hydroxylation was also a determinant of serological reactivity, since an alpha-hydroxylated CMH, only present at the Y clone, was recognized by the hyperimmune sera. In summary, these data indicate that T. cruzi CMHs with non-hydroxylated fatty acids are unable to induce antibody responses in animal hosts, which is reverted by the addition of a sugar residue or an alpha-hydroxyl group. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15727820&query_hl=1 ER - TY - JFULL T1 - Variable significance of low levels of Bcr-Abl transcripts in the peripheral blood after allogeneic stem cell transplant for chronic myeloid leukaemia in chronic phase A1 - O'Shea, DM A1 - Kaeda, J A1 - Szydlo, R A1 - Olavarria, E A1 - Marin, D A1 - Dazzi, F A1 - Perz, JB A1 - Anand, M A1 - Saunders, S A1 - Goldman, JM A1 - Apperley, JF J1 - BONE MARROW TRANSPL Y1 - 2005/03// VL - 35 SN - 0268-3369 SP - S68 EP - S69 ER - TY - JFULL T1 - What causes acute coronary syndromes? Applying Koch's postulates. A1 - Monaco, C A1 - Mathur, A A1 - Martin, JF J1 - Atherosclerosis Y1 - 2005/03// VL - 179 SN - 0021-9150 SP - 1 EP - 15 N2 - The term "acute coronary syndromes" (ACS) is used to describe a heterogeneous spectrum of clinical conditions. This includes myocardial infarction, non-ST-elevation myocardial infarction, and unstable angina. These conditions are linked by a similar constellation of signs and symptoms but not necessarily by a common pathophysiology. They are syndromes. Several different hypotheses exist that have attempted to explain the pathological mechanisms that are involved in these conditions, however, it is not clear whether ACS are caused by variations of a single disease process or by several disease processes. The contribution of both vessel wall- and blood-related factors in the pathogenesis of acute coronary syndromes is herein discussed with the guidance of Koch's postulates. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15721004&query_hl=1 ER - TY - JFULL T1 - Engagement of the p55 TNF-R for different periods of time induces qualitatively distinct pathways of tumour necrosis factor signalling and gene expression profiles in murine T cells A1 - Vagenas, P A1 - Clark, JM A1 - Testar, J A1 - Panesar, M A1 - Udalova, I A1 - Freeman, T A1 - Lyons, P A1 - Cope, AP J1 - ARTHRITIS RES THER Y1 - 2005/02// VL - 7 SN - 1478-6362 SP - S27 EP - S27 ER - TY - JFULL T1 - Role of CCL11 in eosinophilic lung disease during respiratory syncytial virus infection. A1 - Matthews, SP A1 - Tregoning, JS A1 - Coyle, AJ A1 - Hussell, T A1 - Openshaw, PJ J1 - J Virol Y1 - 2005/02// VL - 79 SN - 0022-538X SP - 2050 EP - 2057 N2 - Respiratory syncytial virus (RSV) is a major viral pathogen of infants and the elderly. Significant morbidity is caused by an overexuberant mixed lung cell infiltrate, which is thought to be driven by chemokines. One of the main chemotactic mediators responsible for the movement of eosinophils is CCL11 (eotaxin). Using a mouse model of eosinophilic bronchiolitis induced by RSV, we show here that treatment in vivo with a blocking antibody to CCL11 greatly reduces lung eosinophilia and disease severity. In addition, anti-CCL11 caused a striking inhibition of CD4-T-cell influx and shifted cytokine production away from interleukin-5 without reducing the resistance to viral replication. These results suggest that in addition to influencing eosinophil diapedesis and survival, anti-CCL11 has an action on T cells. These studies strengthen the case for anti-CCL11 treatment of Th2-driven diseases. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15681407&query_hl=1 ER - TY - JFULL T1 - Post-transcriptional regulation of proinflammatory cytokines in arthritis using gene delivery of AU-rich element binding factors A1 - Khoury, M A1 - Apparailly, F A1 - Clark, AR A1 - Plence, P A1 - Noel, D A1 - Jorgensen, C J1 - ARTHRITIS RES THER Y1 - 2005/02// VL - 7 SN - 1478-6362 SP - S28 EP - S28 ER - TY - JFULL T1 - Leg length preservation with pedicled fillet of foot flaps after traumatic amputations. A1 - Ghali, S A1 - Harris, PA A1 - Khan, U A1 - Pearse, M A1 - Nanchahal, J J1 - Plast Reconstr Surg Y1 - 2005/02// VL - 115 SN - 1529-4242 SP - 498 EP - 505 N2 - Six patients with insufficient soft-tissue coverage after lower limb trauma were treated with pedicled fillet of foot flaps to achieve primary stump closure and to preserve leg length. The flaps used were all based on either the posterior tibial neurovascular pedicle, the anterior tibial neurovascular pedicle, or both. Five flaps survived; one patient required conversion of a through-knee to an above-knee amputation and debridement of the flap because of venous thrombosis of the pedicle. In three of the cases, a functional knee joint was preserved. The patients ranged in age from 21 to 54 years, the mean hospital stay was 55.5 days (range, 28 to 76 days), and the mean follow-up time was 14.5 months. Despite an average of 4.3 procedures from initial admission to first discharge and an average of 2.0 postamputation procedures to achieve primary stump healing, all patients have achieved independent mobility with their prosthesis. The advantages of preserving leg length and, where possible, preserving a functional knee joint compensate for repeated procedures on these patients. When planned well, a pedicled fillet of foot flap therefore achieves the aims of amputation, namely, providing primary healing of a sensate, durable, cylindrical stump that is pain-free and preserves maximal leg length. This is achieved with no donor-site morbidity and with no need for microvascular reconstruction. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15692356&query_hl=1 ER - TY - JFULL T1 - Proteomic characterisation of cell contact-dependent macrophage activation A1 - Peirce, MJ A1 - Wait, R A1 - Wu, X A1 - Begum, S A1 - Saklatvala, J A1 - Cope, AP J1 - ARTHRITIS RES THER Y1 - 2005/02// VL - 7 SN - 1478-6362 SP - S29 EP - S29 ER - TY - JFULL T1 - Mesenchymal stem cells induce division arrest anergy of activated T cells A1 - Glennie, SJ A1 - Soeiro, I A1 - Dyson, J A1 - Lam, E A1 - Dazzi, F J1 - BIOL BLOOD MARROW TR Y1 - 2005/02// VL - 11 SN - 1083-8791 SP - 1 EP - 1 ER - TY - JFULL T1 - Collagenases and aggrecanases: understanding the role of non-catalytic domains in cartilage matrix breakdown A1 - Nagase, H A1 - Visse, R A1 - Kashiwagi, M A1 - Gendron, C A1 - Itoh, Y J1 - ARTHRITIS RES THER Y1 - 2005/02// VL - 7 SN - 1478-6362 SP - S3 EP - S3 ER - TY - JFULL T1 - Irradiation influences endothelial cell function in vitro and in vivo: A possible role for IP10 A1 - Cannella, L A1 - Laylor, R A1 - Marelli-Berg, F A1 - Dazzi, F J1 - BIOL BLOOD MARROW TR Y1 - 2005/02// VL - 11 SN - 1083-8791 SP - 45 EP - 45 ER - TY - JFULL T1 - Sustained downregulation of the TCR zeta chain defines a transition from antigen mode to inflammation mode during terminal T-cell differentiation A1 - Zhang, Z A1 - Panesar, M A1 - Amjadi, P A1 - Foey, A A1 - Owen, S A1 - Dazzi, F A1 - Brennan, FM A1 - Cope, AP J1 - ARTHRITIS RES THER Y1 - 2005/02// VL - 7 SN - 1478-6362 SP - S30 EP - S30 ER - TY - JFULL T1 - Release of tumour necrosis factor alpha (TNF alpha) by TNF alpha cleaving enzyme (TACE) in response to septic stimuli in vitro A1 - Robertshaw, HJ A1 - Brennan, FM J1 - BRIT J ANAESTH Y1 - 2005/02// VL - 94 SN - 0007-0912 SP - 222 EP - 228 N2 - Background. Tumour necrosis factor alpha (TNFalpha), in its soluble form (solTNF), has been well described as an important cytokine in inflammatory states including sepsis. The transmembrane precursor of solTNF, membrane-bound TNFalpha (memTNF), is cleaved by TNFalpha cleaving enzyme (TACE), the regulation of which is poorly understood. We hypothesized that the diversity of clinical features seen with sepsis caused by different organisms may be a result of differential regulation of TACE. Therefore, we measured these proteins in models of sepsis using flow cytometric methods that we have developed.Methods. Surface protein expression of memTNF and TACE, and TACE catalytic activity were measured in human monocytes by flow cytometry following cell stimulation by lipopolysaccharide (LPS), zymosan (a yeast cell wall product) or heat-inactivated Neisseria meninigitidis.Results. Unstimulated human monocytes express memTNF on the cell surface (mean fluorescence intensity, MFI 131) and this is down-regulated initially in response to LPS (MFI 57) but then recovers to exceed the resting protein expression (MFI 614). TACE protein is also present on the surface of resting cells (MFI 389) but is catalytically inactive until cell stimulation. Stimulation of monocytes with LPS, zymosan and Neisseria meningitidis produced different patterns of TACE activation with time.Conclusions. The regulation of memTNF by TACE on monocytes is an important regulatory event in the pro-inflammatory cytokine cascade. As monocytes are important in the inflammatory cascade, we suggest that the control of memTNF and TACE activity on monocytes may play a role in the pathophysiology of sepsis. ER - TY - JFULL T1 - Exploring mechanisms of inflammatory hyperalgesia in murine collagen-induced arthritis A1 - Inglis, JJ A1 - Anand, P A1 - Facer, P A1 - Criado, G A1 - Feldmann, M A1 - Williams, RO J1 - ARTHRITIS RES THER Y1 - 2005/02// VL - 7 SN - 1478-6362 SP - S44 EP - S45 ER - TY - JFULL T1 - Activation of antigen-presenting cells by endogenous retroviral RNA A1 - Moyes, D A1 - Sacre, S A1 - Temperton, N A1 - Lundberg, A A1 - Foxwell, B A1 - Venables, P J1 - ARTHRITIS RES THER Y1 - 2005/02// VL - 7 SN - 1478-6362 SP - S12 EP - S13 ER - TY - JFULL T1 - How does tumour necrosis factor uncouple T-cell receptor-induced IL-2 gene expression in murine T cells? A1 - Clark, JM A1 - Aleksiyadis, K A1 - Panesar, M A1 - Cope, AP J1 - ARTHRITIS RES THER Y1 - 2005/02// VL - 7 SN - 1478-6362 SP - S17 EP - S18 ER - TY - JFULL T1 - Citrullinated alpha enolase, a novel citrullinated autoantigen in rheumatoid arthritis, upregulated by chronic inflammation A1 - Kinloch, A A1 - Tatzer, V A1 - Wait, R A1 - Peston, D A1 - Sacre, S A1 - Donatien, P A1 - Moyes, D A1 - Taylor, P A1 - Venables, PJ J1 - ARTHRITIS RES THER Y1 - 2005/02// VL - 7 SN - 1478-6362 SP - S11 EP - S11 ER - TY - JFULL T1 - Synovial hypoxia inducible factor-1 alpha expression is inversely related to tissue oxygen levels in vivo in inflammatory arthritis A1 - Donatien, P A1 - Sandhu, V A1 - Peston, D A1 - Sandisson, A A1 - Taylor, PC J1 - ARTHRITIS RES THER Y1 - 2005/02// VL - 7 SN - 1478-6362 SP - S6 EP - S6 ER - TY - JFULL T1 - Bystander-activated CD4(+) memory lymphocytes: a role in the pathology of rheumatoid arthritis? A1 - Beech, JT A1 - Owen, SH A1 - Green, P A1 - Amjadi, P A1 - Brennan, FM J1 - ARTHRITIS RES THER Y1 - 2005/02// VL - 7 SN - 1478-6362 SP - S52 EP - S52 ER - TY - JFULL T1 - The role of hypoxia in rheumatoid tendon disease A1 - Sivakumar, B A1 - Kang, N A1 - Taylor, P A1 - Paleolog, EM J1 - ARTHRITIS RES THER Y1 - 2005/02// VL - 7 SN - 1478-6362 SP - S7 EP - S7 ER - TY - JFULL T1 - Treatment of collagen-induced arthritis with cholera toxin-treated dendritic cells A1 - Criado, G A1 - Inglis, JJ A1 - Feldmann, M A1 - Williams, RO J1 - ARTHRITIS RES THER Y1 - 2005/02// VL - 7 SN - 1478-6362 SP - S51 EP - S51 ER - TY - JFULL T1 - Expression of E-selectin in coxsackievirus B3 myocarditis A1 - Yeh, JS A1 - Tracy, S A1 - Drescher, KM A1 - Sunde, J A1 - Kono, K A1 - Chapman, N A1 - Boyle, J A1 - Hussell, T A1 - Sennoga, C A1 - Eckersley, R A1 - Blomley, M A1 - Haskard, D A1 - Nihoyannopoulos, P J1 - J AM COLL CARDIOL Y1 - 2005/02/01/ VL - 45 SN - 0735-1097 SP - 142A EP - 142A ER - TY - JFULL T1 - CD44 binding through the hemopexin-like domain is critical for its shedding by membrane-type 1 matrix metalloproteinase. A1 - Suenaga, N A1 - Mori, H A1 - Itoh, Y A1 - Seiki, M J1 - Oncogene Y1 - 2005/01/27/ VL - 24 SN - 0950-9232 SP - 859 EP - 868 N2 - Membrane-type 1 matrix metalloproteinase (MT1-MMP) is a potent modulator of pericellular environment through its proteolytic activity and promotes migration, invasion, and proliferation of tumor cells. During cell migration, MT1-MMP binds to CD44H, a major hyaluronan receptor, through the hemopexin-like (HPX) domain and localizes at the migration front. MT1-MMP is also responsible for shedding CD44H, which supports CD44H-mediated cell migration. In this study, we asked whether the binding of MT1-MMP to CD44H is a prerequisite step for the successive shedding. Deletion of the HPX domain deprived MT1-MMP of its shedding activity. Furthermore, disruption of the CD44H/MT1-MMP complex by overexpressing the HPX fragments resulted in inhibition of the shedding. Thus, the CD44H in the complex appears to be the direct substrate of MT1-MMP for shedding. Interestingly, other members of the MT-MMP family showed varied extents of CD44H shedding. Domain swapping between MT1-MMP and other MT-MMPs revealed that the ability of the HPX domains to bind CD44H is conserved among them. However, the shedding activity was different depending on the catalytic domains. The conserved binding ability of the HPX domains suggests that CD44H may act as a core molecule assembling multiple MT-MMPs on the cell surface. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15558018&query_hl=1 ER - TY - JFULL T1 - Persistent systemic inflammation in unstable angina is largely unrelated to the atherothrombotic burden. A1 - Monaco, C A1 - Rossi, E A1 - Milazzo, D A1 - Citterio, F A1 - Ginnetti, F A1 - D'Onofrio, G A1 - Cianflone, D A1 - Crea, F A1 - Biasucci, LM A1 - Maseri, A J1 - J Am Coll Cardiol Y1 - 2005/01/18/ VL - 45 SN - 0735-1097 SP - 238 EP - 243 N2 - OBJECTIVES: The aim of this study was to assess the relationship between systemic inflammation, atherosclerosis, and thrombosis in two distinct clinical models of atherothrombosis. BACKGROUND: Persistent unstable angina (UA) is commonly associated with coronary thrombosis and persistent systemic inflammation. METHODS: We assessed circulating markers of activation of the thrombotic and fibrinolytic cascades and systemic soluble and cellular markers of inflammation on admission in 40 patients with persisting UA (Braunwald class IIIB; group 1) and 30 patients with Leriche-Fontaine stage IIB-III peripheral artery disease awaiting revascularization (group 2). RESULTS: The extent of atherosclerosis (p < 0.01) and activation of the coagulation system were greater in group 2, which had higher thrombin-antithrombin III complexes and D-dimer levels (2.7 and 24.4 microg/l, respectively), than in group 1 (2.0 microg/l and 12.9 microg/l, p = 0.02 and p = 0.0001, respectively). In contrast, C-reactive protein and interleukin-6 levels were higher in group 1 (7.6 pg/ml and 7.8 pg/ml, respectively) than in group 2 (4.5 pg/ml and 3.0 pg/ml, p < 0.01 and p = 0.03, respectively). Moreover, neutrophil activation was only found in group 1 (neutrophil myeloperoxidase content -4.0 arbitrary units vs. +3.4 arbitrary units in group 2, p < 0.0001). These differences persisted during the initial three days of hospitalization. CONCLUSIONS: Such a large, consistent discrepancy between atherothrombotic burden and systemic inflammation suggests that atherothrombosis, by itself, is an unlikely cause of persisting, recurring UA. An understanding of the primary inflammatory mechanisms of persistent and recurrent coronary instability could open the way to novel therapeutic strategies. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15653021&query_hl=1 ER - TY - JFULL T1 - Models of rheumatoid arthritis. A1 - Williams, RO J1 - Ernst Schering Res Found Workshop Y1 - 2005/// SN - 0947-6075 SP - 89 EP - 117 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15526938&query_hl=1 ER - TY - JFULL T1 - The toll-like receptor-nuclear factor kappaB pathway in rheumatoid arthritis. A1 - Andreakos, E A1 - Sacre, S A1 - Foxwell, BM A1 - Feldmann, M J1 - Front Biosci Y1 - 2005/// VL - 10 SN - 1093-4715 SP - 2478 EP - 2488 N2 - The study of the role cytokines play in the pathogenesis of rheumatoid arthritis (RA) has provided a whole new range of targets for drug development. Many of them (e.g. TNF, IL-1, IL-6, IL-15 and IL-18) are already being targeted in the clinic with success using neutralizing monoclonal antibodies or soluble cytokine receptors. Targeting TNF, in particular, has shown great efficacy in controlling both the inflammation and structural damage of the joints, setting a new gold standard for the treatment of RA. However, what triggers the production of inflammatory cytokines such as TNF in RA remains to be determined. In this article, we review evidence suggesting that the transcription factor Nuclear Factor kappaB (NF-kappaB) is essential for the expression of both inflammatory cytokines and tissue destructive enzymes in RA. Also, we discuss whether Toll-like receptors (TLRs), major receptors involved in pathogen recognition and potent activators of the NF-kappaB pathway, are involved in triggering the inflammatory and joint destructive process in RA and whether they constitute sensible targets for monoclonal antibodies/soluble receptors and small molecule inhibitors. We conclude that although the TLR- NF-kappaB pathway offers ample opportunities for therapeutic intervention, future drugs to be approved will need to match or exceed the efficacy and safety of anti-TNF agents, with safety the most difficult aspect to predict. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15970510&query_hl=1 ER - TY - JFULL T1 - Proteome analysis of serovars Typhimurium and Pullorum of Salmonella enterica subspecies I. A1 - Encheva, V A1 - Wait, R A1 - Gharbia, SE A1 - Begum, S A1 - Shah, HN J1 - BMC Microbiol Y1 - 2005/// VL - 5 SN - 1471-2180 SP - 42 EP - 42 N2 - BACKGROUND: Salmonella enterica subspecies I includes several closely related serovars which differ in host ranges and ability to cause disease. The basis for the diversity in host range and pathogenic potential of the serovars is not well understood, and it is not known how host-restricted variants appeared and what factors were lost or acquired during adaptations to a specific environment. Differences apparent from the genomic data do not necessarily correspond to functional proteins and more importantly differential regulation of otherwise identical gene content may play a role in the diverse phenotypes of the serovars of Salmonella. RESULTS: In this study a comparative analysis of the cytosolic proteins of serovars Typhimurium and Pullorum was performed using two-dimensional gel electrophoresis and the proteins of interest were identified using mass spectrometry. An annotated reference map was created for serovar Typhimurium containing 233 entries, which included many metabolic enzymes, ribosomal proteins, chaperones and many other proteins characteristic for the growing cell. The comparative analysis of the two serovars revealed a high degree of variation amongst isolates obtained from different sources and, in some cases, the variation was greater between isolates of the same serovar than between isolates with different sero-specificity. However, several serovar-specific proteins, including intermediates in sulphate utilisation and cysteine synthesis, were also found despite the fact that the genes encoding those proteins are present in the genomes of both serovars. CONCLUSION: Current microbial proteomics are generally based on the use of a single reference or type strain of a species. This study has shown the importance of incorporating a large number of strains of a species, as the diversity of the proteome in the microbial population appears to be significantly greater than expected. The characterisation of a diverse selection of strains revealed parts of the proteome of S. enterica that alter their expression while others remain stable and allowed for the identification of serovar-specific factors that have so far remained undetected by other methods. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16026608&query_hl=1 ER - TY - JFULL T1 - It's all in the blood: circulating endothelial progenitor cells link synovial vascularity with cardiovascular mortality in rheumatoid arthritis? A1 - Paleolog, E J1 - Arthritis Res Ther Y1 - 2005/// VL - 7 SN - 1478-6362 SP - 270 EP - 272 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16277702&query_hl=1 ER - TY - JFULL T1 - Stimulation via Toll-like receptor 9 reduces Cryptococcus neoformans-induced pulmonary inflammation in an IL-12-dependent manner. A1 - Edwards, L A1 - Williams, AE A1 - Krieg, AM A1 - Rae, AJ A1 - Snelgrove, RJ A1 - Hussell, T J1 - Eur J Immunol Y1 - 2005/01// VL - 35 SN - 0014-2980 SP - 273 EP - 281 N2 - Cytosine-phosphate-guanosine-containing oligodeoxynucleotides (CpG ODN) are important vaccine adjuvants that promote Th1-type immune responses. Cryptococcus neoformans is a serious human pathogen that replicates in the lung but may disseminate systemically leading to meningitis, particularly in immunocompromised individuals. Immunization of susceptible C57BL/6 mice with CpG ODN deviates the immune response from a Th2- toward a Th1-type response following infection with C. neoformans. CpG also induces IL-12, TNF, MCP-1 and macrophage nitric oxide production. CD4(+) and CD8(+) T cells producing IFN-gamma increase in frequency, while those producing IL-5 decrease. More importantly, pulmonary eosinophilia is significantly reduced, an effect that depends on IL-12 and CD8(+) T cells but not NK cells. CpG treatment also reduces the burden of C. neoformans in the lung, an effect that is IL-12-, NK cell- and T cell-independent and probably reflects a direct effect of CpG on pathogen opsonization or an enhancement of macrophage antimicrobial activity. An equivalent beneficial effect is also observed when CpG ODN treatment is delivered during established cryptococcal disease. This is the first study documenting that promotion of lung TLR9 signaling using synthetic agonists enhances host defense. Activation of innate immunity has clear therapeutic potential and may even be beneficial in patients with acquired immune deficiency. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15597328&query_hl=1 ER - TY - JFULL T1 - Pathogenic role of TNFa in rheumatoid arthritis A1 - Larche, M. J. A1 - Sacre, S. M. A1 - Foxwell, B. M. J1 - Drug Discovery Today: Disease Mechanisms Y1 - 2005/// IS - 3 VL - 2 SP - 367 EP - 375 ER - TY - JFULL T1 - Advances in the modulation of cutaneous wound healing and scarring. A1 - Miller, MC A1 - Nanchahal, J J1 - BioDrugs Y1 - 2005/// VL - 19 SN - 1173-8804 SP - 363 EP - 381 N2 - Cutaneous wounds inevitably heal with scars, which can be disfiguring and compromise function. In general, the greater the insult, the worse the scarring, although genetic make up, regional variations and age can influence the final result. Excessive scarring manifests as hypertrophic and keloid scars. At the other end of the spectrum are poorly healing chronic wounds, such as foot ulcers in diabetic patients and pressure sores. Current therapies to minimize scarring and accelerate wound healing rely on the optimization of systemic conditions, early wound coverage and closure of lacerations, and surgical incisions with minimal trauma to the surrounding skin. The possible benefits of topical therapies have also been assessed. Further major improvements in wound healing and scarring require an understanding of the molecular basis of this process. Promising strategies for modulating healing include the local administration of platelet derived growth factor (PDGF)-BB to accelerate the healing of chronic ulcers, and increasing the relative ratio of transforming growth factor (TGF)beta-3 to TGFbeta-1 and TGFbeta-2 in order to minimize scarring. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16392889&query_hl=1 ER - TY - JFULL T1 - Nitric oxide causes activation of JNK and suppression of Akt in insulin-secreting cells A1 - Storling, J A1 - Binzer, J A1 - Andersson, AK A1 - Tonnesen, M A1 - Billestrup, N A1 - Sandler, S A1 - Mandrup-Poulsen, T J1 - DIABETOLOGIA Y1 - 2005/01// VL - 48 SN - 0012-186X SP - A38 EP - A38 ER - TY - JFULL T1 - Identification of citrullinated alpha-enolase as a candidate autoantigen in rheumatoid arthritis. A1 - Kinloch, A A1 - Tatzer, V A1 - Wait, R A1 - Peston, D A1 - Lundberg, K A1 - Donatien, P A1 - Moyes, D A1 - Taylor, PC A1 - Venables, PJ J1 - Arthritis Res Ther Y1 - 2005/// VL - 7 SN - 1478-6362 SP - R1421 EP - R1429 N2 - Antibodies against citrullinated proteins are highly specific for rheumatoid arthritis (RA), but little is understood about their citrullinated target antigens. We have detected a candidate citrullinated protein by immunoblotting lysates of monocytic and granulocytic HL-60 cells treated with peptidylarginine deiminase. In an initial screen of serum samples from four patients with RA and one control, a protein of molecular mass 47 kDa from monocytic HL-60s reacted with sera from the patients, but not with the serum from the control. Only the citrullinated form of the protein was recognised. The antigen was identified by tandem mass spectrometry as alpha-enolase, and the positions of nine citrulline residues in the sequence were determined. Serum samples from 52 patients with RA and 40 healthy controls were tested for presence of antibodies against citrullinated and non-citrullinated alpha-enolase by immunoblotting of the purified antigens. Twenty-four sera from patients with RA (46%) reacted with citrullinated alpha-enolase, of which seven (13%) also recognised the non-citrullinated protein. Six samples from the controls (15%) reacted with both forms. Alpha-enolase was detected in the RA joint, where it co-localised with citrullinated proteins. The presence of antibody together with expression of antigen within the joint implicates citrullinated alpha-enolase as a candidate autoantigen that could drive the chronic inflammatory response in RA. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16277695&query_hl=1 ER - TY - JFULL T1 - Molecular diversity and thrombotic risk in Protein S deficiency: The PROSIT study A1 - Biguzzi, E A1 - Razzari, C A1 - Lane, DA A1 - Castaman, G A1 - Cappellari, A A1 - Bucciarelli, P A1 - Fontana, G A1 - Margaglione, M A1 - D'Andrea, G A1 - Simmonds, RE A1 - Rezende, SM A1 - Preston, R A1 - Prisco, D A1 - Faioni, EM A1 - Protein S Italian Team J1 - HUM MUTAT Y1 - 2005/// VL - 25 SN - 1059-7794 SP - 259 EP - 269 N2 - The Protein S Italian Team (PROSIT) enrolled 79 protein S (PS) deficient families and found 38 PROS1 variations (19 novel) in 53 probands. Of these, 23 variants were selected for expression in vitro, to evaluate their role as possible causative variants. Transient expression showed high secretion levels (> 75%) for three variants, which were considered neutral. Seven missense and five nonsense variants showed low (:!; 11%) expression levels and were classified as severe defects. Intermediate expression was observed for eight variants, which were evaluated by factor Va inactivation assay in order to be globally classified as severe or intermediate. Based on the cumulative data, the hazard ratio associated with causative variants was 4.9 (95% CI: 1.4-17-7) for deep vein thrombosis and/or pulmonary embolism, 5.1 (95% CI: 1.1-23.9) for superficial thrombophlebitis, and 4.8 (95% CI: 1.8-13.0) for any venous thrombosis. The hazard ratio for deep vein thrombosis and/or pulmonary embolism in carriers of severe defects only was 7.4 (95% CI: 1.6-24.1). PROSIT showed that dysfunctional variants causing PS deficiency are more common than expected and confirmed that PS deficiency is associated with increased thrombotic risk, although risk assessment is complicated by molecular heterogeneity. (C) 2005 Wiley-Liss, Inc. ER - TY - JFULL T1 - Load balancing by changing the graph connectivity on heterogeneous clusters A1 - Munasinghe, K A1 - Wait, R J1 - LECT NOTES COMPUT SC Y1 - 2005/// VL - 3470 SN - 0302-9743 SP - 1040 EP - 1047 N2 - This paper examines the problem of adapting parallel applications on a cluster of workstations. The cluster is assumed to be a heterogeneous, multi-user computing environment so that, efficient load balancing within the application must take external factors into account. At any time the users of the network are competing for resources. Performance of a particular processor, as a component in the parallel (message passing) computation, depends on both static factors, such as the processor hardware, and dynamic factors, such as the system load and the activities of other users. For each processor, the external factors can be condensed into a single parameter, the load index, which is a normalised measure of the current spare capacity of the processor available to the application.Numerical experiments show the efficiency of the load balancing strategies on a finite element application with a domain decomposition and the effect on overall computation time. ER - TY - JFULL T1 - Anti-TNF therapy: where have we got to in 2005? A1 - Feldmann, M A1 - Brennan, FM A1 - Foxwell, BM A1 - Taylor, PC A1 - Williams, RO A1 - Maini, RN J1 - J Autoimmun Y1 - 2005/// VL - 25 Suppl SN - 0896-8411 SP - 26 EP - 28 N2 - The blockade of TNF has had significant impact on the therapy of a number of chronic autoimmune diseases. In this chapter we review the concepts leading up to this therapy in rheumatoid arthritis (RA), how it spreads into other autoimmune diseases, and how greater understanding of its use has led to augmented therapeutic benefit. There are still many limitations, but the prospects for the future are intriguing. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16260118&query_hl=1 ER - TY - JFULL T1 - Relationships between the N-glycan structures and biological activities of recombinant human erythropoietins produced using different culture conditions and purification procedures. A1 - Yuen, CT A1 - Storring, PL A1 - Tiplady, RJ A1 - Izquierdo, M A1 - Wait, R A1 - Gee, CK A1 - Gerson, P A1 - Lloyd, P A1 - Cremata, JA J1 - Adv Exp Med Biol Y1 - 2005/// VL - 564 SN - 0065-2598 SP - 141 EP - 142 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=16400821&query_hl=1 ER - TY - JFULL T1 - Stabilization of IFN-gamma mRNA by MAPK p38 in IL-12- and IL-18-stimulated human NK cells. A1 - Mavropoulos, A A1 - Sully, G A1 - Cope, AP A1 - Clark, AR J1 - Blood Y1 - 2005/01/01/ VL - 105 SN - 0006-4971 SP - 282 EP - 288 N2 - The rapid induction of interferon-gamma (IFN-gamma) by innate cytokines such as interleukin 12 (IL-12) and IL-18 is critical for immunity against infectious pathogens. We investigated the molecular mechanisms underlying this response. IL-12 and IL-18 rapidly and synergistically induced the secretion of IFN-gamma by freshly purified human peripheral blood lymphocytes. At early time points, IFN-gamma was expressed almost exclusively by natural killer cells and in both CD56bright and CD56dim subpopulations. Mitogen-activated protein kinase p38 was activated strongly by IL-18 and weakly by IL-12 in natural killer cells but was not activated by either cytokine in T cells. The expression of IFN-gamma mRNA and protein was dose-dependently blocked by SB203580, a specific inhibitor of mitogen-activated protein kinase p38, which also caused a dramatic destabilization of IFN-gamma mRNA. The 3' untranslated region (UTR) of IFN-gamma mRNA conferred p38 responsiveness to a heterologous reporter mRNA. Therefore, the synergistic induction of IFN-gamma by IL-12 and IL-18 in natural killer cells is mediated at least in part by p38-dependent and 3' UTR-mediated stabilization of IFN-gamma mRNA. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15345584&query_hl=1 ER - TY - JFULL T1 - Association between baseline radiographic damage and improvement in physical function after treatment of patients with rheumatoid arthritis. A1 - Breedveld, FC A1 - Han, C A1 - Bala, M A1 - van der Heijde, D A1 - Baker, D A1 - Kavanaugh, AF A1 - Maini, RN A1 - Lipsky, PE J1 - Ann Rheum Dis Y1 - 2005/01// VL - 64 SN - 0003-4967 SP - 52 EP - 55 N2 - OBJECTIVES: To identify factors associated with poor physical function in rheumatoid arthritis and to assess whether baseline joint damage has an impact on improvement in physical function during infliximab treatment. METHODS: 428 patients with active rheumatoid arthritis despite methotrexate treatment received methotrexate alone or with infliximab (3 mg/kg or 10 mg/kg every four or eight weeks) for 54 weeks (the ATTRACT trial). Data on clinical outcomes and physical function (assessed by the health assessment questionnaire (HAQ)) were collected. Structural damage was assessed using the van der Heijde modification of the Sharp score. Odds ratios (OR) for factors associated with severe functional disability (HAQ > or =2.0) at baseline were estimated using multiple logistic regression analyses, and baseline factors related to the change in physical function after treatment at week 54 were determined. RESULTS: Baseline radiographic scores were correlated with baseline HAQ scores. After adjustment for demographic characteristics in the logistic regression model, baseline disease activity scores, radiological joint damage, fatigue, and morning stiffness were found to be associated with severe functional disability (HAQ >2.0), with OR values of 2.00 (1.53 to 2.63), 1.82 (1.15 to 2.87), 1.19 (1.05 to 1.34), and 1.07 (1.01 to 1.13), respectively. In multiple linear regression analysis, physical disability, joint damage, and fatigue at baseline were correlated with less improvement in physical function after treatment. Infliximab treatment was associated with greater improvement in physical function. CONCLUSIONS: Greater joint damage at baseline was associated with poorer physical function at baseline and less improvement in physical function after treatment, underlining the importance of early intervention to slow the progression of joint destruction. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15286005&query_hl=1 ER - TY - JFULL T1 - RHAMM, a receptor for hyaluronan-mediated motility, compensates for CD44 in inflamed CD44-knockout mice: a different interpretation of redundancy. A1 - Nedvetzki, S A1 - Gonen, E A1 - Assayag, N A1 - Reich, R A1 - Williams, RO A1 - Thurmond, RL A1 - Huang, JF A1 - Neudecker, BA A1 - Wang, FS A1 - Wang, FS A1 - Turley, EA A1 - Naor, D J1 - Proc Natl Acad Sci U S A Y1 - 2004/12/28/ VL - 101 SN - 0027-8424 SP - 18081 EP - 18086 N2 - We report here that joint inflammation in collagen-induced arthritis is more aggravated in CD44-knockout mice than in WT mice, and we provide evidence for molecular redundancy as a causal factor. Furthermore, we show that under the inflammatory cascade, RHAMM (receptor for hyaluronan-mediated motility), a hyaluronan receptor distinct from CD44, compensates for the loss of CD44 in binding hyaluronic acid, supporting cell migration, up-regulating genes involved with inflammation (as assessed by microarrays containing 13,000 cDNA clones), and exacerbating collagen-induced arthritis. Interestingly, we further found that the compensation for loss of the CD44 gene does not occur because of enhanced expression of the redundant gene (RHAMM), but rather because the loss of CD44 allows increased accumulation of the hyaluronic acid substrate, with which both CD44 and RHAMM engage, thus enabling augmented signaling through RHAMM. This model enlightens several aspects of molecular redundancy, which is widely discussed in many scientific circles, but the processes are still ill defined. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15596723&query_hl=1 ER - TY - JFULL T1 - Low density lipoprotein receptor-related protein mediates endocytic clearance of pro-MMP-2.TIMP-2 complex through a thrombospondin-independent mechanism. A1 - Emonard, H A1 - Bellon, G A1 - Troeberg, L A1 - Berton, A A1 - Robinet, A A1 - Henriet, P A1 - Marbaix, E A1 - Kirkegaard, K A1 - Patthy, L A1 - Eeckhout, Y A1 - Nagase, H A1 - Hornebeck, W A1 - Courtoy, PJ J1 - J Biol Chem Y1 - 2004/12/24/ VL - 279 SN - 0021-9258 SP - 54944 EP - 54951 N2 - The low density lipoprotein receptor-related protein (LRP) mediates the endocytic clearance of various proteinases and proteinase.inhibitor complexes, including thrombospondin (TSP)-dependent endocytosis of matrix metalloproteinase (MMP)-2 (or gelatinase A), a key effector of extracellular matrix remodeling and cancer progression. However, the zymogen of MMP-2 (pro-MMP-2) mostly occurs in tissues as a complex with the tissue inhibitor of MMPs (TIMP-2). Here we show that clearance of the pro-MMP-2.TIMP-2 complex is also mediated by LRP, because addition of receptor-associated protein (RAP), a natural LRP ligand antagonist, inhibited endocytosis and lysosomal degradation of (125)I-pro-MMP-2.TIMP-2. Both TIMP-2 and the pro-MMP-2 collagen-binding domain independently competed for endocytosis of (125)I-pro-MMP-2.TIMP-2 complex. Surface plasmon resonance studies indicated that pro-MMP-2, TIMP-2, and pro-MMP-2.TIMP-2 directly interact with LRP in the absence of TSP. LRP-mediated endocytic clearance of (125)I-pro-MMP-2 was inhibited by anti-TSP antibodies and accelerated upon complexing with TSP-1, but these treatments had no effect on (125)I-pro-MMP-2.TIMP-2 uptake. This implies that mechanisms of clearance by LRP of pro-MMP-2 and pro-MMP-2.TIMP-2 complex are different. Interestingly, RAP did not inhibit binding of (125)I-pro-MMP-2.TIMP-2 to the cell surface. We conclude that clearance of pro-MMP-2.TIMP-2 complex is a TSP-independent two-step process, involving (i) initial binding to the cell membrane in a RAP-insensitive manner and (ii) subsequent LRP-dependent (RAP-sensitive) internalization and degradation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15489233&query_hl=1 ER - TY - JFULL T1 - Low density lipoprotein receptor-related protein mediates endocytic clearance of Pro-MMP-2 center dot TIMP-2 complex through a thrombospondin-independent mechanism A1 - Emonard, H A1 - Bellon, G A1 - Troeberg, L A1 - Berton, A A1 - Robinet, A A1 - Henriet, P A1 - Marbaix, E A1 - Kirkegaard, K A1 - Patthy, L A1 - Eeckhout, Y A1 - Nagase, H A1 - Hornebeck, W A1 - Courtoy, PJ J1 - J BIOL CHEM Y1 - 2004/12/24/ VL - 279 SN - 0021-9258 SP - 54944 EP - 54951 N2 - The low density lipoprotein receptor-related protein (LRP) mediates the endocytic clearance of various proteinases and proteinase . inhibitor complexes, including thrombospondin (TSP)-dependent endocytosis of matrix metalloproteinase (MMP)-2 ( or gelatinase A), a key effector of extracellular matrix remodeling and cancer progression. However, the zymogen of MMP-2 (pro-MMP-2) mostly occurs in tissues as a complex with the tissue inhibitor of MMPs (TIMP-2). Here we show that clearance of the pro-MMP-2 . TIMP-2 complex is also mediated by LRP, because addition of receptor-associated protein ( RAP), a natural LRP ligand antagonist, inhibited endocytosis and lysosomal degradation of I-125-pro-MMP-2 . TIMP-2. Both TIMP-2 and the pro-MMP-2 collagen-binding domain independently competed for endocytosis of I-125-pro-MMP-2 . TIMP-2 complex. Surface plasmon resonance studies indicated that pro-MMP-2, TIMP-2, and pro-MMP-2 . TIMP- 2 directly interact with LRP in the absence of TSP. LRP-mediated endocytic clearance of I-125-pro-MMP-2 was inhibited by anti-TSP antibodies and accelerated upon complexing with TSP-1, but these treatments had no effect on I-125-pro-MMP-2 . TIMP- 2 uptake. This implies that mechanisms of clearance by LRP of pro-MMP-2 and pro-MMP-2 . TIMP- 2 complex are different. Interestingly, RAP did not inhibit binding of I-125-pro-MMP-2 . TIMP- 2 to the cell surface. We conclude that clearance of pro-MMP-2 . TIMP- 2 complex is a TSP-independent two-step process, involving (i) initial binding to the cell membrane in a RAP-insensitive manner and (ii) subsequent LRP-dependent (RAP-sensitive) internalization and degradation. ER - TY - JFULL T1 - Innate imprinting by the modified heat-labile toxin of Escherichia coli (LTK63) provides generic protection against lung infectious disease. A1 - Williams, AE A1 - Edwards, L A1 - Humphreys, IR A1 - Snelgrove, R A1 - Rae, A A1 - Rappuoli, R A1 - Hussell, T J1 - J Immunol Y1 - 2004/12/15/ VL - 173 SN - 0022-1767 SP - 7435 EP - 7443 N2 - In a healthy individual, the lung contains few lymphoid cells. However, amplified immune responses, as exemplified during lung infection, can cause extensive tissue damage. We have previously demonstrated that one lung infection modulates the immunopathological outcome to a subsequent unrelated pathogen. Mimicking heterologous immunity may provide a means of enhancing both innate and acquired immunity. We now show that prior lung administration of a modified heat-labile toxin from Escherichia coli (LTK63) enhances immunity to respiratory syncytial virus, influenza virus, and the fungus Cryptococcus neoformans. Treatment with LTK63 decreased lung inflammation and tissue damage and improved the ability to resolve the infection. APCs expressing the activation markers MHC class II, CD80, and CD40 increased in number in the lung. LTK63 treatment increased the pathogen-specific IgA response in the nasal mucosa and simultaneously decreased inflammatory cytokine production (IFN-gamma and TNF-alpha) after infection. The number of activated CD8(+)CD44(+) T cells and the respiratory syncytial virus- or influenza-specific CD8-proliferative responses increased, although the total inflammatory infiltrate was reduced. LTK63 treatment matured lung APCs (LTK63 prevented efficient presentation of whole OVA to DO11.10 cells, whereas OVA peptide presentation was unaffected), enhanced immunity in both a Th1 and Th2 environment, was long lasting, and was not pathogen or host strain specific; the protective effects were partially independent of T and B cells. Innate imprinting by toxin-based immunotherapeutics may provide generic protection against infectious disease in the lung, without the need for coadministered pathogen-specific Ag. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15585869&query_hl=1 ER - TY - JFULL T1 - How the gut links innate and adaptive immunity. A1 - Rumbo, M A1 - Anderle, P A1 - Didierlaurent, A A1 - Sierro, F A1 - Debard, N A1 - Sirard, JC A1 - Finke, D A1 - Kraehenbuhl, JP J1 - Ann N Y Acad Sci Y1 - 2004/12// VL - 1029 SN - 0077-8923 SP - 16 EP - 21 N2 - Mucosal surfaces represent the main sites in which environmental microorganisms and antigens interact with the host. Sentinel cells, including epithelial cells, lumenal macrophages, and intraepithelial dendritic cells, continuously sense the environment and coordinate defenses for the protection of mucosal tissues. The mucosal epithelial cells are crucial actors in coordinating defenses. They sense the outside world and respond to environmental signals by releasing chemokines and cytokines that recruit inflammatory and immune cells to control potential infectious agents and to attract cells able to trigger immune responses. Among immune cells, dendritic cells (DC) play a key role in controlling adaptive immune responses, due to their capacity to internalize foreign materials and to present antigens to naive T and B lymphocytes, locally or in draining organized lymphoid tissues. Immune cells recruited in epithelial tissues can, in turn, act upon the epithelial cells and change their phenotype in a process referred to as epithelial metaplasia. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15681739&query_hl=1 ER - TY - JFULL T1 - A novel mechanism for TNF-alpha regulation by p38 MAPK: involvement of NF-kappa B with implications for therapy in rheumatoid arthritis. A1 - Campbell, J A1 - Ciesielski, CJ A1 - Hunt, AE A1 - Horwood, NJ A1 - Beech, JT A1 - Hayes, LA A1 - Denys, A A1 - Feldmann, M A1 - Brennan, FM A1 - Foxwell, BM J1 - J Immunol Y1 - 2004/12/01/ VL - 173 SN - 0022-1767 SP - 6928 EP - 6937 N2 - TNF-alpha is a key factor in a variety of inflammatory diseases. This study examines the role of p38 MAPK in the regulation of TNF-alpha in primary human cells relevant to inflammation, e.g., macrophages and rheumatoid synovial cells. Using a dominant negative variant (D168A) of p38 MAPK and a kinase inhibitor, SB203580, we confirm in primary human macrophages that p38 MAPK regulates TNF-alpha production using a posttranscriptional mechanism requiring the 3' untranslated region of the gene. However, in LPS-activated primary human macrophages we also detect a second previously unidentified mechanism, the p38 MAPK modulation of TNF-alpha transcription. This is mediated through p38 MAPK regulation of NF-kappaB. Interestingly this mechanism was not observed in rheumatoid synovial cells. Importantly however, the dominant negative mutant of p38 MAPK, but not SB203580 was effective at inhibiting spontaneous TNF-alpha production in these ex vivo rheumatoid synovial cell cultures. These data indicate there are potential major differences in the role of p38 MAPK in inflammatory signaling that have a bearing on the use of this kinase as a target for therapy. These results indicate despite disappointing results with p38 MAPK inhibitors in the clinic, this kinase is a valid target in rheumatoid disease. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15557189&query_hl=1 ER - TY - JFULL T1 - Coregulation of fibronectin signaling and matrix contraction by tenascin-C and syndecan-4. A1 - Midwood, KS A1 - Valenick, LV A1 - Hsia, HC A1 - Schwarzbauer, JE J1 - Mol Biol Cell Y1 - 2004/12// VL - 15 SN - 1059-1524 SP - 5670 EP - 5677 N2 - Syndecan-4 is a ubiquitously expressed heparan sulfate proteoglycan that modulates cell interactions with the extracellular matrix. It is transiently up-regulated during tissue repair by cells that mediate wound healing. Here, we report that syndecan-4 is essential for optimal fibroblast response to the three-dimensional fibrin-fibronectin provisional matrix that is deposited upon tissue injury. Interference with syndecan-4 function inhibits matrix contraction by preventing cell spreading, actin stress fiber formation, and activation of focal adhesion kinase and RhoA mediated-intracellular signaling pathways. Tenascin-C is an extracellular matrix protein that regulates cell response to fibronectin within the provisional matrix. Syndecan-4 is also required for tenascin-C action. Inhibition of syndecan-4 function suppresses tenascin-C activity and overexpression of syndecan-4 circumvents the effects of tenascin-C. In this way, tenascin-C and syndecan-4 work together to control fibroblast morphology and signaling and regulate events such as matrix contraction that are essential for efficient tissue repair. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15483051&query_hl=1 ER - TY - JFULL T1 - Tumour necrosis factor alpha as a therapeutic target for immune-mediated inflammatory diseases. A1 - Taylor, PC A1 - Williams, RO A1 - Feldmann, M J1 - Curr Opin Biotechnol Y1 - 2004/12// VL - 15 SN - 0958-1669 SP - 557 EP - 563 N2 - Preclinical studies have identified and validated tumour necrosis factor alpha (TNFalpha) as a key disease molecule and therapeutic target for immunotherapeutic intervention in many immune-mediated inflammatory diseases. Clinical indications include rheumatoid arthritis, Crohn's disease, ankylosing spondylitis and psoriasis. Recent clinical findings indicate that many chronic inflammatory disorders share certain pathogenic pathways, whereas others are limited to particular disease phenotypes. Better understanding of these pathogenic pathways will inform the development of new therapeutic approaches leading to more complete and sustained disease remissions. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15560982&query_hl=1 ER - TY - JFULL T1 - Development of a solid-phase assay for analysis of matrix metalloproteinase activity. A1 - Lauer-Fields, JL A1 - Nagase, H A1 - Fields, GB J1 - J Biomol Tech Y1 - 2004/12// VL - 15 SN - 1524-0215 SP - 305 EP - 316 N2 - Proteases play fundamentally important roles in normal physiology and disease pathology. Methods for detection of active proteolysis may greatly aid in the diagnosis of disease progression, and suggest modes of therapeutic intervention. Most assays for proteolytic potential are limited by a lack of specificity and/or quantification. We have developed a solid-phase activity assay for members of the matrix metalloproteinase (MMP) family that is specific and can be used to quantify active enzyme concentration. The assay has two principal components: a capture antibody that immobilizes the MMP without perturbing the enzyme active site, and a fluorescence resonance energy transfer substrate for monitoring proteolysis at low enzyme concentrations. The assay was standardized for MMP-1, MMP-3, MMP-13, and MMP-14. The efficiency of the assay was found to be critically dependent upon the quality of the antibodies, the use of substrates exhibiting high specific activities for the enzymes, and enzyme samples that are fresh. The assay was applied to studies of constitutive and induced MMP activity in human melanoma cells. Analysis of several melanoma cell lines, and comparison with prior studies, correlated higher constitutive MMP-13 activity with higher levels of the cell surface receptor CD44. Ligands to two different melanoma cell surface receptors (the alpha2beta1 integrin or CD44) were found to induce different proteolytic profiles, suggesting that the extracellular matrix can modulate melanoma invasion. Overall, the solid-phase MMP activity assay was found to be valuable for analysis of protease activity in cellular environments. The solid-phase assay is suitably flexible to allow studies of virtually any proteolytic enzyme for which appropriate substrates and antibodies are available. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15585827&query_hl=1 ER - TY - JFULL T1 - Emerging approaches for the therapy of autoimmune and chronic inflammatory disease. A1 - Cope, AP A1 - Feldmann, M J1 - Curr Opin Immunol Y1 - 2004/12// VL - 16 SN - 0952-7915 SP - 780 EP - 786 N2 - Progress in defining protein and gene signatures that characterize autoimmune-mediated inflammatory diseases has uncovered a large number of potential therapeutic targets. Preclinical data from rodent models can be generated rapidly, as can data from the genetic crosses of gene-deficient mice on autoimmune-susceptible backgrounds. But humans are not the same as mice, and however robust preclinical data might appear, therapeutic intervention in patients with autoimmune disease remains the definitive experiment. Several studies published in the past year have tested paradigms of autoimmune disease in clinical trials. Recent therapeutic approaches for targeting B-cell subsets and co-stimulatory pathways are described here in detail. It is our belief that the future of immunotherapy in the clinic will depend to some extent upon the availability of biomarkers for defining biological signatures of immune function in vivo. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15511673&query_hl=1 ER - TY - JFULL T1 - Pain reduction and lack of psychotropic effects with ajulemic acid: comment on the article by Sumariwalla et al - Reply A1 - Sumariwalla, PF A1 - Feldmann, M A1 - Gallily, R A1 - Mechoulam, R A1 - Tchilibon, S A1 - Fride, E J1 - ARTHRITIS RHEUM Y1 - 2004/12// VL - 50 SN - 0004-3591 SP - 4079 EP - 4080 ER - TY - JFULL T1 - Is targeting Toll-like receptors and their signaling pathway a useful therapeutic approach to modulating cytokine-driven inflammation? A1 - Andreakos, E A1 - Foxwell, B A1 - Feldmann, M J1 - Immunol Rev Y1 - 2004/12// VL - 202 SN - 0105-2896 SP - 250 EP - 265 N2 - Cytokine-driven inflammation and tissue destruction is a common theme of chronic inflammatory diseases such as rheumatoid arthritis, Crohn's disease, ulcerative colitis, psoriasis, chronic obstructive pulmonary disease, and atherosclerosis. Research over the last two decades demonstrated the importance of cytokines that are not only expressed chronically but also are capable of signaling at sites of chronic inflammation. Cytokines thus regulate major pathological processes that include inflammation, angiogenesis, tissue remodeling, and fibrosis. This research led to the identification of key cytokines involved in these processes, two of which, tumor necrosis factor-alpha and interleukin-1, have also been successfully targeted in the clinic. However, what triggers and maintains cytokine gene expression in chronic inflammation remains a mystery. In this article, we review current progress in the understanding of cytokine-driven inflammation and discuss current evidence implicating Toll-like receptors (TLRs), recently identified as the receptors recognizing self versus non-self molecular patterns, in the regulation of cytokine-driven inflammation. Whether targeting TLRs and their downstream signaling pathway will prove to be a successful approach for the treatment of these devastating diseases remains to be determined. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15546398&query_hl=1 ER - TY - JFULL T1 - Identification of selective basophil chemoattractants in human nasal polyps as insulin-like growth factor-1 and insulin-like growth factor-2. A1 - Hartnell, A A1 - Heinemann, A A1 - Conroy, DM A1 - Wait, R A1 - Sturm, GJ A1 - Caversaccio, M A1 - Jose, PJ A1 - Williams, TJ J1 - J Immunol Y1 - 2004/11/15/ VL - 173 SN - 0022-1767 SP - 6448 EP - 6457 N2 - In a search for novel leukocyte chemoattractants at sites of allergic inflammation, we found basophil-selective chemoattractant activity in extracts of human nasal polyps. The extracts were fractionated by reverse phase HPLC, and the resulting fractions were tested for leukocyte-stimulating activity using sensitive shape change assays. The basophil-selective activity detected was not depleted by a poxvirus CC-chemokine-binding protein affinity column. This activity was further purified by HPLC, and proteins in the bioactive fractions were analyzed by tandem electrospray mass spectrometry. Insulin-like growth factor-2 (IGF-2) was identified in these HPLC fractions, and the basophil-stimulating activity was inhibited by an anti-IGF-2-neutralizing Ab. Recombinant IGF-2 induced a substantial shape change response in basophils, but not eosinophils, neutrophils, or monocytes. IGF-2 stimulated chemokinesis of basophils, but not eosinophils or neutrophils, and synergized with eotaxin-1/CCL11 in basophil chemotaxis. IGF-2 also caused up-regulation of basophil CD11b expression and inhibited apoptosis, but did not stimulate degranulation or Ca(2+) flux. Recombinant IGF-1 exhibited similar basophil-selective effects as IGF-2, and both growth factors were detected in nasal polyp extracts by ELISA. This is the first demonstration of chemokinetic factors that increase the motility of basophils, but do not act on other granulocytes or monocytes. IGF-1 and IGF-2 could play a role in the selective recruitment of basophils in vivo. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15528386&query_hl=1 ER - TY - JFULL T1 - Combination of infliximab and methotrexate therapy for early rheumatoid arthritis: a randomized, controlled trial. A1 - St Clair, EW A1 - van der Heijde, DM A1 - Smolen, JS A1 - Maini, RN A1 - Bathon, JM A1 - Emery, P A1 - Keystone, E A1 - Schiff, M A1 - Kalden, JR A1 - Wang, B A1 - Dewoody, K A1 - Weiss, R A1 - Baker, D A1 - Active-Controlled Study of Patients Receiving Infliximab for the Treatment of Rheumatoid Arthritis of Early Onset Study Group J1 - Arthritis Rheum Y1 - 2004/11// VL - 50 SN - 0004-3591 SP - 3432 EP - 3443 N2 - OBJECTIVE: To compare the benefits of initiating treatment with methotrexate (MTX) and infliximab (anti-tumor necrosis factor alpha [anti-TNFalpha] monoclonal antibody) with those of MTX treatment alone in patients with rheumatoid arthritis (RA) of < or =3 years' duration. METHODS: RA patients were eligible if they had active disease and no prior treatment with MTX or a TNFalpha inhibitor. One thousand forty-nine patients were randomly assigned in a 4:5:5 ratio to 3 treatment groups: MTX-placebo, MTX-3 mg/kg infliximab, and MTX-6 mg/kg infliximab. MTX dosages were rapidly escalated to 20 mg/week, and infliximab or placebo infusions were given at weeks 0, 2, and 6, and every 8 weeks thereafter through week 46. RESULTS: At week 54, the median percentage of American College of Rheumatology improvement (ACR-N) was higher for the MTX-3 mg/kg infliximab and MTX-6 mg/kg infliximab groups than for the MTX-placebo group (38.9% and 46.7% versus 26.4%, respectively; P < 0.001 for both comparisons). Patients in the MTX-3 mg/kg infliximab and MTX-6 mg/kg infliximab groups also showed less radiographic progression than those receiving MTX alone (mean +/- SD changes in van der Heijde modification of the total Sharp score at week 54: 0.4 +/- 5.8 and 0.5 +/- 5.6 versus 3.7 +/- 9.6, respectively; P < 0.001 for each comparison). In addition, physical function improved significantly more in the MTX-3 mg/kg infliximab and MTX-6 mg/kg infliximab groups than in the MTX-placebo group. Infliximab therapy was associated with a significantly higher incidence of serious infections, especially pneumonia. CONCLUSION: For patients with active RA in its early stages, combination therapy with MTX and infliximab provides greater clinical, radiographic, and functional benefits than treatment with MTX alone. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15529377&query_hl=1 ER - TY - JFULL T1 - The role of tumour necrosis factor (TNF-alpha) in experimental autoimmune uveoretinitis (EAU). A1 - Dick, AD A1 - Forrester, JV A1 - Liversidge, J A1 - Cope, AP J1 - Prog Retin Eye Res Y1 - 2004/11// VL - 23 SN - 1350-9462 SP - 617 EP - 637 N2 - The pleiotropic cytokine tumour necrosis factor-alpha (TNF-alpha) is released from cells that include macrophages and T-cells during inflammatory responses, orchestrating the initiation of further leucocytic infiltration via adhesion molecule upregulation, dendritic cell maturation and survival, macrophage activation and driving Th1 T-cells responses within tissues. Exposure to TNF also plays a role in maintaining tissue homeostasis, particularly relating to resident cell responses of both microglia and retinal pigment epithelium. Depending on the balance between duration and dose of TNF exposure, an environment where full expression of inflammatory and autoimmune responses within tissues may occur. In experimental autoimmune uveoretinitis (EAU), increased tissue concentrations of TNF facilitate the on-going T-cell effector responses and macrophage activation. These are responsible for targeted and bystander tissue damage and can be suppressed by anti-TNF therapies, in particular, those directed at the p55 TNF receptor. The ability to suppress disease experimentally has led to the successful translation of anti-TNF therapy for treatment of uveitis in cohort studies and phase I/II trials where, additionally, altered peripheral blood CD4(+) T-cell profiles can be demonstrated following each treatment. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15388077&query_hl=1 ER - TY - JFULL T1 - Interleukin-10 suppression of myeloid cell activation--a continuing puzzle. A1 - Williams, LM A1 - Ricchetti, G A1 - Sarma, U A1 - Smallie, T A1 - Foxwell, BM J1 - Immunology Y1 - 2004/11// VL - 113 SN - 0019-2805 SP - 281 EP - 292 N2 - Efforts to identify the signal transduction pathways used by interleukin-10 (IL-10) have resulted in limited success. The anti-inflammatory effects elicited by IL-10, and the mechanisms by which these are mediated, are still relatively unknown. Understanding the signalling mechanisms behind the suppression of cytokine expression by IL-10 could be of potential therapeutic interest. Although the consensus is that the Janus kinase, Jak1, as well as the signal transducer and activator of transcription STAT3 are central, much controversy exists about the participation and roles of many other signalling pathways targeted by IL-10. The mechanisms of cytokine suppression proposed by various groups have included transcriptional, post-transcriptional and post-translational regulation of IL-10 target genes; nevertheless no unifying model has emerged thus far. Here we would like to highlight novel findings and discuss their implications in the context of current understanding of IL-10 signalling. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15500614&query_hl=1 ER - TY - JFULL T1 - Secreted frizzled-related protein-1 inhibits RANKL-dependent osteoclast formation. A1 - Häusler, KD A1 - Horwood, NJ A1 - Chuman, Y A1 - Fisher, JL A1 - Ellis, J A1 - Martin, TJ A1 - Rubin, JS A1 - Gillespie, MT J1 - J Bone Miner Res Y1 - 2004/11// VL - 19 SN - 0884-0431 SP - 1873 EP - 1881 N2 - We determined that sFRP-1 mRNA was differentially expressed by osteoblast/stromal cell lines and that sFRP-1 neutralizing antibodies and siRNA complementary to sFRP-1 coding sequence enhanced, while recombinant sFRP-1 inhibited, osteoclast formation. In studying the mechanism of action for sFRP-1, we found that sFRP-1 could bind recombinant RANKL. These results suggest potential cross-talk between Wnt and RANKL pathways. INTRODUCTION: Osteoclast formation in normal bone remodeling requires the presence of osteoblast lineage cells that express RANKL and macrophage-colony-stimulating factor (M-CSF), which interact with their cognate receptors on the osteoclast precursor. We identified secreted Frizzled-related protein-1 (sFRP-1), which is known to bind to Wnt and inhibit the Wnt signaling pathway, as an osteoblast-derived factor that impinges on osteoclast formation and activity. MATERIALS AND METHODS: Differential display of mRNA from osteoblast lineage cell lines established sFRP-1 to be highly expressed in an osteoclast supporting cell line. sFRP-1 expression in bone was determined by in situ hybridization, and the effects of sFRP-1 on osteoclast formation were determined using a neutralizing antibody, siRNA, for sFRP-1 and recombinant protein. RESULTS: In situ hybridization revealed sFRP-1 mRNA expression in osteoblasts and chondrocytes in murine bone. sFRP-1 mRNA expression could be elevated in calvarial primary osteoblasts in response to prostaglandin E2 (PGE2) or interleukin (IL)-11, whereas many other osteotropic agents (e.g., IL-1, IL-6, calcitrol, parathyroid hormone) were without any effect. In vitro assays of osteoclast formation established sFRP-1 to be an inhibitor of osteoclast formation. Neutralizing antibodies against sFRP-1 enhanced TRACP+ mononuclear and multinuclear osteoclast formation (3- and 2-fold, respectively) in co-cultures of murine osteoblasts with spleen cells, whereas siRNA complementary to sFRP-1 coding sequence significantly enhanced osteoclast formation in co-cultures of KUSA O (osteoblast/stromal cell line) and bone marrow cells, cultured in the presence of PGE2 and 1,25(OH)2 vitamin D3. Recombinant sFRP-1 dose-dependently inhibited osteoclast formation in osteoblast/spleen co-cultures, RANKL + M-CSF-treated splenic cultures, and RANKL-treated RAW264.7 cell cultures, indicating a direct action of sFRP-1 on hematopoietic cells. Consistent with this, sFRP-1 was found to bind to RANKL in ELISAs. CONCLUSION: sFRP-1 is expressed by osteoblasts and inhibits osteoclast formation. While sFRP-1 activity might involve the blocking of endogenous Wnt signaling, our results suggest that, alternatively, it could be because of direct binding to RANKL. This study describes a new mechanism whereby osteoblasts regulate osteoclastogenesis through the expression and release of sFRP-1. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15476588&query_hl=1 ER - TY - JFULL T1 - Chondrogenic differentiation of murine embryonic stem cells: effects of culture conditions and dexamethasone. A1 - Tanaka, H A1 - Murphy, CL A1 - Murphy, C A1 - Kimura, M A1 - Kawai, S A1 - Polak, JM J1 - J Cell Biochem Y1 - 2004/10/15/ VL - 93 SN - 0730-2312 SP - 454 EP - 462 N2 - Pluripotent embryonic stem (ES) cells have the capability to differentiate to various cell types and may represent an alternative cell source for the treatment of cartilage defects. Here, we show that differentiation of ES cells toward the chondrogenic lineage can be enhanced by altering the culture conditions. Chondrogenesis was observed in intact embryoid body (EB) cultures, as detected by an increase in mRNA levels for aggrecan and Sox9 genes. Collagen IIB mRNA, the mature chondrocyte-specific splice variant, was absent at day 5, but appeared at later time points. Dexamethasone treatment of alginate-encapsulated EB cultures did not have a strong chondrogenic effect. Nor was chondrogenesis enhanced by alginate encapsulation compared to simple plating of EBs. However, disruption of day 5 EBs and culture as a micromass or pelleted mass, significantly enhanced the expression of the cartilage marker gene collagen type II and the transcription factor Sox9 compared to all other treatments. Histological and immunohistochemical analysis of pellet cultures revealed cartilage-like tissue characterized by metachromatically stained extracellular matrix and type II collagen immunoreactivity, indicative of chondrogenesis. These findings have potentially important implications for cartilage tissue engineering, since they may enable the increase in differentiated cell numbers needed for the in vitro development of functional cartilaginous tissue suitable for implantation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15372628&query_hl=1 ER - TY - JFULL T1 - Proteomic analysis of articular cartilage shows increased type II collagen synthesis in osteoarthritis and expression of inhibin betaA (activin A), a regulatory molecule for chondrocytes. A1 - Hermansson, M A1 - Sawaji, Y A1 - Bolton, M A1 - Alexander, S A1 - Wallace, A A1 - Begum, S A1 - Wait, R A1 - Saklatvala, J J1 - J Biol Chem Y1 - 2004/10/15/ VL - 279 SN - 0021-9258 SP - 43514 EP - 43521 N2 - We show that proteomic analysis can be applied to study cartilage pathophysiology. Proteins secreted by articular cartilage were analyzed by two-dimensional SDS-PAGE and mass spectrometry. Cartilage explants were cultured in medium containing [35S]methionine/cysteine to radiolabel newly synthesized proteins. To resolve the cartilage proteins by two-dimensional electrophoresis, it was necessary to remove the proteoglycan aggrecan by precipitation with cetylpyridinium chloride. 50-100 radiolabeled protein spots were detected on two-dimensional gels of human cartilage cultures. Of 170 silver-stained proteins identified, 19 were radiolabeled, representing newly synthesized gene products. Most of these were known cartilage constituents. Several nonradiolabeled cartilage proteins were also detected. The secreted protein pattern of explants from 12 osteoarthritic joints (knee, hip, and shoulder) and 14 nonosteoarthritic adult joints were compared. The synthesis of type II collagen was strongly up-regulated in osteoarthritic cartilage. Normal adult cartilage synthesized little or no type II collagen in contrast to infant and juvenile cartilage. Potential regulatory molecules novel to cartilage were identified; pro-inhibin betaA and processed inhibin betaA (which dimerizes to activin A) were produced by all the osteoarthritic samples and half of the normals. Connective tissue growth factor and cytokine-like protein C17 (previously only identified as an mRNA) were also found. Activin induced the tissue inhibitor for metalloproteinases-1 in human chondrocytes. Its expression was induced in isolated chondrocytes by growth factors or interleukin-1. We conclude that type II collagen synthesis in articular cartilage is down-regulated at skeletal maturity and reactivated in osteoarthritis in attempted repair and that activin A may be an anabolic factor in cartilage. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15292256&query_hl=1 ER - TY - JFULL T1 - Advanced glycation end products upregulate angiogenic and pro-inflammatory cytokine production in human monocyte/macrophages. A1 - Pertynska-Marczewska, M A1 - Kiriakidis, S A1 - Wait, R A1 - Beech, J A1 - Feldmann, M A1 - Paleolog, EM J1 - Cytokine Y1 - 2004/10/07/ VL - 28 SN - 1043-4666 SP - 35 EP - 47 N2 - Glucose can react non-enzymatically with amino groups of, for example, proteins, to yield derivatives termed advanced glycation end products (AGE), which contribute to many chronic progressive diseases associated with microvascular complications. The study aimed to determine the effect of AGE-modified albumin on THP-1 cells and human monocyte-derived macrophages. Bovine serum albumin (BSA) or human serum albumin (HSA), modified by glucose-derived AGE, was prepared by incubation with glucose for differing periods of time. Alternatively, BSA was incubated with sodium cyanoborohydride and glyoxylic acid to produce N(epsilon)-(carboxymethyl)lysine-modified BSA (CML-BSA). Stimulation for 24h of THP-1 cells with BSA, incubated for 6-8 weeks with glucose, induced significant VEGF release. Human monocyte-derived macrophages stimulated with extensively glycated HSA also showed significant VEGF release, as well as upregulation of IL-8 production, incubation for 6h with extensively glycated HSA increased release of TNFalpha and expression of tissue factor. Finally, addition of CML-BSA resulted in significant induction of TNFalpha and VEGF release. We demonstrate that a range of different methods of glycation of BSA and HSA, including CML-BSA, resulted in the induction of VEGF, TNFalpha, IL-8 and expression of tissue factor, according to length of stimulation and different glycation products used, suggesting that AGE-induced activation of macrophages may contribute to vascular complications by regulation of angiogenic, inflammatory and pro-coagulant processes. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15341924&query_hl=1 ER - TY - JFULL T1 - The involvement of AU-rich element-binding proteins in p38 mitogen-activated protein kinase pathway-mediated mRNA stabilisation. A1 - Dean, JL A1 - Sully, G A1 - Clark, AR A1 - Saklatvala, J J1 - Cell Signal Y1 - 2004/10// VL - 16 SN - 0898-6568 SP - 1113 EP - 1121 N2 - The p38 mitogen-activated protein kinase (MAPK) pathway plays an important role in the post-transcriptional regulation of inflammatory genes. p38 has been found to regulate both the translation and the stability of inflammatory mRNAs. The mRNAs regulated by p38 share common AU-rich elements (ARE) present in their 3'-untranslated regions. AREs act as mRNA instability determinants but also confer stabilisation of the mRNA by the p38 pathway. In recent years, AREs have shown to be binding sites for numerous proteins including HuR, TTP, AUF1, AUF2, FBP1, FBP2 (KSRP), TIA-1, and TIAR. However, it is unclear which protein is responsible for mRNA stabilisation by p38. This review gives an overview of the major ARE-binding proteins and discusses reasons for and against their involvement in p38-mediated mRNA stabilisation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15240006&query_hl=1 ER - TY - JFULL T1 - Detection and mapping of widespread intermolecular protein disulfide formation during cardiac oxidative stress using proteomics with diagonal electrophoresis. A1 - Brennan, JP A1 - Wait, R A1 - Begum, S A1 - Bell, JR A1 - Dunn, MJ A1 - Eaton, P J1 - J Biol Chem Y1 - 2004/10/01/ VL - 279 SN - 0021-9258 SP - 41352 EP - 41360 N2 - Regulation of protein function by reversible cysteine-targeted oxidation can be achieved by multiple mechanisms, such as S-glutathiolation, S-nitrosylation, sulfenic acid, sulfinic acid, and sulfenyl amide formation, as well as intramolecular disulfide bonding of vicinal thiols. Another cysteine oxidation state with regulatory potential involves the formation of intermolecular protein disulfides. We utilized two-dimensional sequential non-reducing/reducing SDS-PAGE (diagonal electrophoresis) to investigate intermolecular protein disulfide formation in adult cardiac myocytes subjected to a series of interventions (hydrogen peroxide, S-nitroso-N-acetylpenicillamine, doxorubicin, simulated ischemia, or metabolic inhibition) that alter the redox status of the cell. More detailed experiments were undertaken with the thiol-specific oxidant diamide (5 mm), a concentration that induces a mild non-injurious oxidative stress. This increase in cellular oxidation potential caused global intermolecular protein disulfide formation in cytosolic, membrane, and myofilament/cytoskeletal compartments. A large number of proteins that undergo these associations were identified using liquid chromatography-mass spectrometry/mass spectrometry. These associations, which involve metabolic and antioxidant enzymes, structural proteins, signaling molecules, and molecular chaperones, were confirmed by assessing "shifts" on non-reducing immunoblots. The observation of widespread protein-protein disulfides indicates that these oxidative associations are likely to be fundamental in how cells respond to redox changes. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15292244&query_hl=1 ER - TY - JFULL T1 - Recombinant respiratory syncytial virus lacking secreted glycoprotein G is attenuated, non-pathogenic but induces protective immunity. A1 - Maher, CF A1 - Hussell, T A1 - Blair, E A1 - Ring, CJ A1 - Openshaw, PJ J1 - Microbes Infect Y1 - 2004/10// VL - 6 SN - 1286-4579 SP - 1049 EP - 1055 N2 - Respiratory syncytial virus (RSV) causes intense pulmonary inflammatory responses in some infected infants. The surface attachment protein 'G' of RSV has membrane-bound and secreted forms and shows homology to the CX3C chemokine fractalkine. Using recombinant techniques, we generated replication-competent recombinant clonal RSV expressing normal G proteins ('rRSV') or only the membrane-bound form of G ('Gmem rRSV'). Both recombinants grew well in HEp-2 cells, but after primary intranasal infection in mice, pulmonary Gmem rRSV replication was reduced tenfold compared to parental or rRSV; moreover, CCL2 and CCL5 production was greatly reduced and no apparent disease or pulmonary cellular infiltration was observed. However, Gmem rRSV-infected mice developed good antibody responses and were fully protected against subsequent intranasal challenge with parental virus. Even in mice sensitized to G by cutaneous infection with recombinant vaccinia expressing G, intranasal challenge with Gmem rRSV caused insignificant disease. We conclude that secreted G is a key viral product assisting virus replication in vivo, enhancing CCL2 and CCL5 production and promoting illness. Engineered RSV mutants lacking the ability to secrete G are thus promising vaccine candidates. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15380773&query_hl=1 ER - TY - JFULL T1 - Proteomic analysis of the anterior cingulate cortex in schizophrenia, bipolar disorder and major depressive disorder: Evidence for metabolic disturbances A1 - Pennington, K A1 - Beasley, CL A1 - Wait, R A1 - Dunn, M A1 - Cotter, D J1 - AM J MED GENET B Y1 - 2004/09/15/ VL - 130B SN - 1552-4841 SP - 121 EP - 122 ER - TY - JFULL T1 - Matrix metalloproteinase triple-helical peptidase activities are differentially regulated by substrate stability. A1 - Minond, D A1 - Lauer-Fields, JL A1 - Nagase, H A1 - Fields, GB J1 - Biochemistry Y1 - 2004/09/14/ VL - 43 SN - 0006-2960 SP - 11474 EP - 11481 N2 - Matrix metalloproteinases (MMPs) are involved in physiological remodeling as well as pathological destruction of tissues. The turnover of the collagen triple-helical structure has been ascribed to several members of the MMP family, but the determinants for collagenolytic specificity have not been identified. The present study has compared the triple-helical peptidase activities of MMP-1 and MMP-14 (membrane-type 1 MMP; MT1-MMP). The ability of each enzyme to efficiently hydrolyze the triple helix was quantified using chemically synthesized fluorogenic triple-helical substrates that, via addition of N-terminal alkyl chains, differ in their thermal stabilities. One series of substrates was modeled after a collagenolytic MMP consensus cleavage site from types I-III collagen, while the other series had a single substitution in the P(1)' subsite of the consensus sequence. The substitution of Cys(4-methoxybenzyl) for Leu in the P(1)' subsite was greatly favored by MMP-14 but disfavored by MMP-1. An increase in substrate triple-helical thermal stability led to the decreased ability of the enzyme to cleave such substrates, but with a much more pronounced effect for MMP-1. Increased thermal stability was detrimental to enzyme turnover of substrate (k(cat)), but not binding (K(M)). Activation energies were considerably lower for MMP-14 hydrolysis of triple-helical substrates compared with MMP-1. Overall, MMP-1 was found to be less efficient at processing triple-helical structures than MMP-14. These results demonstrate that collagenolytic MMPs have subtle differences in their abilities to hydrolyze triple helices and may explain the relative collagen specificity of MMP-1. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15350133&query_hl=1 ER - TY - JFULL T1 - Mucosal immunisation of murine neonates using whole cell and acellular Pertussis vaccines. A1 - Hale, C A1 - Humphreys, IR A1 - Hussell, T A1 - Bowe, F A1 - Clare, S A1 - Pickard, D A1 - Preston, A A1 - Del Giudice, G A1 - Dougan, G J1 - Vaccine Y1 - 2004/09/09/ VL - 22 SN - 0264-410X SP - 3595 EP - 3602 N2 - Groups of neonatal mice were immunised with different mucosal vaccines based on acellular (Pertactin antigen) or whole cell (inactivated Bordetella pertussis with Diphtheria and Tetanus toxoid) Pertussis vaccines, using Escherichia coli heat-labile enterotoxin (LT) as a mucosal adjuvant. Neonatal mice tolerated mucosal vaccination well and a significant cellular infiltrate was detected in the lungs of mice receiving mucosal vaccines compared to PBS controls. This infiltrate included B lymphocytes, gammadelta T cells and interferon-gamma producing T cells. Neonatal mice, in contrast to adult mice, responded poorly in terms of the production of serum antibody to Pertussis antigens delivered mucosally, although they were able to mount an anti-Tetanus response to those vaccines harbouring Tetanus toxoid and whole cell Pertussis antigen. Neonatal mice immunised with Pertactin or whole cell Pertussis antigen together with LT were protected against virulent B. pertussis challenge. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15315838&query_hl=1 ER - TY - JFULL T1 - Attenuated poxviruses expressing a synthetic HIV protein stimulate HLA-A2-restricted cytotoxic T-cell responses. A1 - Didierlaurent, A A1 - Ramirez, JC A1 - Gherardi, M A1 - Zimmerli, SC A1 - Graf, M A1 - Orbea, HA A1 - Pantaleo, G A1 - Wagner, R A1 - Esteban, M A1 - Kraehenbuhl, JP A1 - Sirard, JC J1 - Vaccine Y1 - 2004/09/03/ VL - 22 SN - 0264-410X SP - 3395 EP - 3403 N2 - Efficient HIV vaccines have to trigger cell-mediated immunity directed against various viral antigens. However little is known about the breadth of the response induced by vaccines carrying multiple proteins. Here, we report on the immunogenicity of a construct harbouring a fusion of the HIV-1 IIIB gag, pol and nef genes (gpn) designed for optimal safety and equimolar expression of the HIV proteins. The attenuated poxviruses, MVA and NYVAC, harbouring the gpn construct, induced potent immune responses in conventional mice characterised by stimulation of Gpn-specific IFN-gamma-producing cells and cytotoxic T cells. In HLA-A2 transgenic mice, recombinant MVA elicited cytotoxic responses against epitopes recognised in most HLA-A2+ HIV-1-infected individuals. We also found that the MVA vaccine triggered the in vitro expansion of peripheral blood cells isolated from a HIV-1-seropositive patient and with similar specificity as found in immunised HLA-A2 transgenic mice. In conclusion, the synthetic HIV polyantigen Gpn delivered by MVA is immunogenic, efficiently processed and presented by human MHC class I molecules. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15308364&query_hl=1 ER - TY - JFULL T1 - A novel citrullinated autoantigen, p47cit, in rheumatoid arthritis. A1 - Kinloch, AJ A1 - Tatzer, V A1 - Wait, R A1 - Moyes, D A1 - Venables, P J1 - ARTHRITIS RHEUM Y1 - 2004/09// VL - 50 SN - 0004-3591 SP - S522 EP - S523 ER - TY - JFULL T1 - Comparison of ultrasonographic measures of synovitis and radiographic assessment of structural damage over two years in a randomised, controlled trial of infliximab therapy in erosive early rheumatoid arthritis. A1 - Taylor, PC A1 - Steuer, A A1 - Gruber, J A1 - McClinton, C A1 - Cosgrove, D A1 - Blomley, M A1 - Wagner, CL A1 - Marsters, PA A1 - Maini, RN J1 - ARTHRITIS RHEUM Y1 - 2004/09// VL - 50 SN - 0004-3591 SP - S402 EP - S402 ER - TY - JFULL T1 - Heme oxygenase 1 expression induced by IL-10 requires STAT-3 and phosphoinositol-3 kinase and is inhibited by lipopolysaccharide. A1 - Ricchetti, GA A1 - Williams, LM A1 - Foxwell, BM J1 - J Leukoc Biol Y1 - 2004/09// VL - 76 SN - 0741-5400 SP - 719 EP - 726 N2 - Heme-oxygenase 1 (HO-1) is a stress-response protein with anti-inflammatory activity. This study has examined the regulation of HO-1 expression by the anti-inflammatory factor, interleukin (IL)-10 and whether HO-1 could account for the function of the cytokine. IL-10-induced expression of HO-1 required the activation of signal transducer and activator of transcription (STAT)-3 but not p38 mitogen-activated protein kinase. However, expression of HO-1 also required the activation of the phosphatidylinositol-3 kinase pathway, a signaling mechanism not required for the anti-inflammatory activity of IL-10. Moreover, induction of HO-1 expression was not restricted to IL-10, as IL-6, a cytokine known to activate STAT-3, could also induce the protein. In human macrophages, lipopolysaccharide inhibited HO-1 expression induced by IL-10. Also, inhibition of HO-1 activity by the specific inhibitor zinc-II-protoporphyrin-IX had no effect on the anti-inflammatory function of IL-10. In summary, although IL-10 does regulate HO-1 expression, it does not appear to play a significant role in the anti-inflammatory activity of the cytokine. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15240748&query_hl=1 ER - TY - JFULL T1 - Protection against the early acute phase of Cryptosporidium parvum infection conferred by interleukin-4-induced expression of T helper 1 cytokines. A1 - McDonald, SA A1 - O'Grady, JE A1 - Bajaj-Elliott, M A1 - Notley, CA A1 - Alexander, J A1 - Brombacher, F A1 - McDonald, V J1 - J Infect Dis Y1 - 2004/09/01/ VL - 190 SN - 0022-1899 SP - 1019 EP - 1025 N2 - Immunity to Cryptosporidium parvum infection involves a T helper (Th) 1 response with interferon (IFN)- gamma and interleukin (IL)-12 activity, but the role of Th2 cytokines, such as IL-4, is unclear. Around the peak of infection, production of oocysts in IL-4-deficient and IL-4 receptor alpha -deficient neonatal BALB/c mice was greater than that in wild-type (wt) mice. Susceptibility to infection was increased or decreased, respectively, in wt mice treated with anti-IL-4 neutralizing antibodies or recombinant IL-4. Excretion of oocysts by IFN- gamma -deficient mice was unaffected by treatment with anti-IL-4, indicating that IL-4 stimulated IFN- gamma activity. Early during infection, wt mice had increased intestinal expression of IFN- gamma and IL-12 mRNA, compared with IL-4-deficient mice. Intestinal IL-4 was detected by Western blotting in wt mice 24 h after infection but not in uninfected control mice. These findings suggest that, early during C. parvum infection of BALB/c mice, there is production of IL-4 that promotes Th1-mediated immunity. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15295711&query_hl=1 ER - TY - JFULL T1 - Glycolytic enzymes on neuronal membranes are candidate autoantigens in post-streptococcal neuropsychiatric disorders A1 - Dale, RC A1 - Candler, PM A1 - Church, AJ A1 - Wait, R A1 - Pocock, JM A1 - Giovannoni, G J1 - MOVEMENT DISORD Y1 - 2004/09// VL - 19 SN - 0885-3185 SP - S33 EP - S33 ER - TY - JFULL T1 - Modulating angiogenesis: more vs less. A1 - Sivakumar, B A1 - Harry, LE A1 - Paleolog, EM J1 - JAMA Y1 - 2004/08/25/ VL - 292 SN - 1538-3598 SP - 972 EP - 977 N2 - The concept of manipulation of the vascular bed to either increase or decrease the number of blood vessels has attracted considerable interest. This review focuses on angiogenesis as a therapeutic target, particularly in the context of cancer and arthritis, as well as on promoting angiogenesis in cardiovascular disease and the healing of bone fractures. Although once touted almost as a panacea for treatment of tumors, as well as other diseases associated with angiogenesis, such as diabetic retinopathy or rheumatoid arthritis, it is now clear that such enthusiasm was somewhat premature. Similarly, some clinical trials of therapeutic angiogenesis for the management of cardiovascular disease have been disappointing. Nevertheless, this exciting field of research holds promise for more targeted therapies. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15328330&query_hl=1 ER - TY - JFULL T1 - Collagenase unwinds triple-helical collagen prior to peptide bond hydrolysis. A1 - Chung, L A1 - Dinakarpandian, D A1 - Yoshida, N A1 - Lauer-Fields, JL A1 - Fields, GB A1 - Visse, R A1 - Nagase, H J1 - EMBO J Y1 - 2004/08/04/ VL - 23 SN - 0261-4189 SP - 3020 EP - 3030 N2 - Breakdown of triple-helical interstitial collagens is essential in embryonic development, organ morphogenesis and tissue remodelling and repair. Aberrant collagenolysis may result in diseases such as arthritis, cancer, atherosclerosis, aneurysm and fibrosis. In vertebrates, it is initiated by collagenases belonging to the matrix metalloproteinase (MMP) family. The three-dimensional structure of a prototypic collagenase, MMP-1, indicates that the substrate-binding site of the enzyme is too narrow to accommodate triple-helical collagen. Here we report that collagenases bind and locally unwind the triple-helical structure before hydrolyzing the peptide bonds. Mutation of the catalytically essential residue Glu200 of MMP-1 to Ala resulted in a catalytically inactive enzyme, but in its presence noncollagenolytic proteinases digested collagen into typical 3/4 and 1/4 fragments, indicating that the MMP-1(E200A) mutant unwinds the triple-helical collagen. The study also shows that MMP-1 preferentially interacts with the alpha2(I) chain of type I collagen and cleaves the three alpha chains in succession. Our results throw light on the basic mechanisms that control a wide range of biological and pathological processes associated with tissue remodelling. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15257288&query_hl=1 ER - TY - JFULL T1 - Co-stimulation: novel methods for preventing viral-induced lung inflammation. A1 - Hussell, T A1 - Snelgrove, R A1 - Humphreys, IR A1 - Williams, AE J1 - Trends Mol Med Y1 - 2004/08// VL - 10 SN - 1471-4914 SP - 379 EP - 386 N2 - Respiratory infections cause significant morbidity and mortality worldwide. Although an immune response is required to eliminate respiratory pathogens, if unchecked, it can damage surrounding tissues and block primary lung function. Based on our knowledge of immune T-cell activation, there are several pathways to which immune intervention could be applied. However, relatively few interventions target only those immune cells that are responding to antigens. OX40 and 4-1BB are members of the tumour necrosis factor receptor family and are expressed on the surface of T cells in several inflammatory conditions. Recently, the inhibition of OX40 has proved beneficial during influenza virus infection. This review highlights the recent advances in the manipulation of such molecules and how they have been applied to inflammatory conditions that are caused by viruses in the lung. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15310458&query_hl=1 ER - TY - JFULL T1 - Identification of major heparin-binding proteins in plasma using electrophoresis and mass spectrometry A1 - Killeen, R A1 - Wait, R A1 - Begum, S A1 - Gray, E A1 - Mulloy, B J1 - INT J EXP PATHOL Y1 - 2004/08// VL - 85 SN - 0959-9673 SP - A69 EP - A69 ER - TY - JFULL T1 - The p38 MAP kinase pathway as a therapeutic target in inflammatory disease. A1 - Saklatvala, J J1 - Curr Opin Pharmacol Y1 - 2004/08// VL - 4 SN - 1471-4892 SP - 372 EP - 377 N2 - The p38 MAPK signalling pathway plays an important role in inflammation and other physiological processes. Specific inhibitors of p38 alpha and beta MAPK block production of the major inflammatory cytokines (i.e. tumour necrosis factor-alpha and interleukin-1) and other proteins (e.g. cyclooxygenase-2), and are anti-inflammatory in animal models of disease. A major function of the pathway is post-transcriptional control of inflammatory gene expression. Many of the mRNAs are unstable (or untranslatable) because of AU-rich elements in the 3'untranslated region. Signalling in the p38 pathway counteracts these and stabilizes the mRNAs by preventing their otherwise rapid de-adenylation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15251131&query_hl=1 ER - TY - JFULL T1 - New approaches to therapeutic immunomodulation for immune-mediated inflammatory disorders. A1 - Taylor, PC A1 - Feldmann, M J1 - Curr Opin Pharmacol Y1 - 2004/08// VL - 4 SN - 1471-4892 SP - 368 EP - 371 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15251130&query_hl=1 ER - TY - JFULL T1 - IL-10 gene therapy is therapeutic for dextran sodium sulfate-induced murine colitis A1 - Lindsay, JO A1 - Sandison, A A1 - Cohen, P A1 - Brennan, FM A1 - Hodgson, HJF J1 - DIGEST DIS SCI Y1 - 2004/08// VL - 49 SN - 0163-2116 SP - 1327 EP - 1334 N2 - The transfer of genes encoding immunoregulatory proteins is a promising new strategy in the treatment of intestinal inflammation. Previous work has demonstrated that daily systemic interleukin (IL)-10 therapy is able to prevent disease onset in animal models of colitis but is not sufficient to treat established disease. This study investigates the therapeutic efficacy of an adenovirus encoding IL-10 (AdvmuIL-10) in the treatment of experimental colitis. Colitis was induced in BALB/c mice by the addition of dextran sodium sulfate to the drinking water for 7 days. A single systemic injection of AdvmuIL-10, empty cassette vector (Adv0), or saline vehicle was administered on day 4 after the onset of colitis. The addition of DSS to the drinking water led to an acute, dose-dependent colitis. A single injection of AdvmuIL-10 led to a marked reduction in both stool markers of inflammation (IL- 1beta, IL-6, and TNFRII) and serum IL-6. Furthermore, the histological colitis score was significantly reduced in mice receiving AdvmuIL-10 compared to controls (4.9 +/- 1.1 Vs 9.1 +/- 1.2, respectively; P < 0.05). A single systemic injection of AdvmuIL-10 is therapeutic in mice with established DSS colitis. Gene therapy strategies using adenoviral vectors encoding IL- 10 may prove to be a potent therapy for chronic inflammation of the colon such as Crohn's disease. ER - TY - JFULL T1 - Induction of interleukin-1 in articular cartilage by explantation and cutting. A1 - Gruber, J A1 - Vincent, TL A1 - Hermansson, M A1 - Bolton, M A1 - Wait, R A1 - Saklatvala, J J1 - Arthritis Rheum Y1 - 2004/08// VL - 50 SN - 0004-3591 SP - 2539 EP - 2546 N2 - OBJECTIVE: To investigate the effect of explantation and fine cutting of articular cartilage upon intracellular inflammatory signaling pathways and expression of interleukin-1 (IL-1). METHODS: Cartilage from porcine metacarpophalangeal joints was cultured in serum-free medium. Tissue extracts were examined for ERK activation by phosphorylated-Western blotting, for JNK and p38 MAPK activity by kinase assay, and for IkappaBalpha. IL-1alpha and IL-1beta messenger RNA (mRNA) was measured by reverse transcriptase-polymerase chain reaction. IL-1 activity was measured by the induction of serum amyloid A protein in cultured chondrocytes. RESULTS: All 3 MAPKs (p38, JNK, and ERK) were rapidly activated upon dissection and explantation of the cartilage. IL-1alpha and IL-1beta mRNA was also induced: the speed and magnitude of induction were increased if the explants had been finely cut. IL-1 activity that could be inhibited by IL-1 receptor antagonist or antibodies to IL-1alpha was found in extracts of explants cultured for 20 hours or lysates of cells isolated from them. This activity was likely due to intracellular proIL-1alpha that was not secreted. ProIL-1beta would not be detected because it is biologically inactive. The mechanism of inflammatory signaling pathway activation underlying the induction of IL-1 is unknown. CONCLUSION: Explantation and cutting of articular cartilage activates intracellular inflammatory signaling pathways and induces expression of mRNA for IL-1alpha and IL-1beta. Biologically active IL-1alpha protein was detectable in cartilage lysates and was probably intracellular proIL-1alpha. We were unable to show that IL-1 was secreted by chondrocytes. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15334468&query_hl=1 ER - TY - JFULL T1 - Imatinib inhibits the activation and proliferation of normal T lymphocytes in vitro. A1 - Cwynarski, K A1 - Laylor, R A1 - Macchiarulo, E A1 - Goldman, J A1 - Lombardi, G A1 - Melo, JV A1 - Dazzi, F J1 - Leukemia Y1 - 2004/08// VL - 18 SN - 0887-6924 SP - 1332 EP - 1339 N2 - The ABL tyrosine kinase inhibitor imatinib mesylate is highly effective in the treatment of CML and is increasingly used in the stem cell transplantation (SCT) setting. Since ABL-dependent intracellular signaling molecules are involved in T-cell activation, imatinib may affect T-cell responses in vivo, thus affecting T-cell function in CML patients, disrupting immune reconstitution after allogeneic SCT and/or impeding the graft-versus-leukemia effect. Here we demonstrate that imatinib inhibits PHA-induced proliferation of normal peripheral blood mononuclear cells at in vitro concentrations (1-5 micromol/l) representative of the pharmacological doses used therapeutically in vivo. The effect is not dependent on antigen-presenting cells because CD3/CD28-induced T-cell stimulation was similarly inhibited by imatinib. Dose-dependent inhibition of the proliferative response of purified CD8+ and CD4+ T lymphocytes to anti-CD3/CD28 was similarly observed and associated with reduction in IFN-gamma production. The inhibitory effect could not be ascribed to an increased rate of apoptosis but the expression of activation markers on CD3+ T cells was significantly reduced in the presence of imatinib (1-5 micromol/L). Inhibition of T-cell proliferation was reversible after removal of the drug from the cultures. Thus, imatinib inhibits T-cell proliferation in vitro, an effect that is APC-independent, reversible, and does not involve apoptosis induction. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15190258&query_hl=1 ER - TY - JFULL T1 - The stability of tristetraprolin mRNA is regulated by mitogen-activated protein kinase p38 and by tristetraprolin itself. A1 - Tchen, CR A1 - Brook, M A1 - Saklatvala, J A1 - Clark, AR J1 - J Biol Chem Y1 - 2004/07/30/ VL - 279 SN - 0021-9258 SP - 32393 EP - 32400 N2 - Tristetraprolin (TTP) is an mRNA-destabilizing protein that negatively regulates the expression of proinflammatory mediators such as tumor necrosis factor alpha, granulocyte/macrophage colony-stimulating factor, and cyclooxygenase 2. Here we investigate the regulation of TTP expression in the mouse macrophage cell line RAW264.7. We show that TTP mRNA is expressed in a biphasic manner following stimulation of cells with lipopolysaccharide and that the second phase of expression, like the first, is dependent on mitogen-activated protein kinase (MAPK) p38. MAPK p38 acts through a downstream kinase to stabilize TTP mRNA, and this stabilization is mediated by an adenosine/uridine-rich region at the 3'-end of the TTP 3'-untranslated region. Hence TTP is post-transcriptionally regulated in a similar manner to several proinflammatory genes. We also demonstrate that TTP is able to bind to its own 3'-untranslated region and negatively regulate its own expression, forming a feedback loop to limit expression levels. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15187092&query_hl=1 ER - TY - JFULL T1 - Adenoviral gene transfer into cells from diseased tissue as a tool to investigate the regulation of inflammatory and destructive mediators in the osteoarthritis synovium A1 - Bondeson, J A1 - Evans, A A1 - Lauder, S A1 - Amos, N A1 - Feldmann, M J1 - ANN RHEUM DIS Y1 - 2004/07// VL - 63 SN - 0003-4967 SP - 64 EP - 64 ER - TY - JFULL T1 - Minor histocompatibility antigens and stem cell transplantation. A1 - Laylor, R A1 - Cannella, L A1 - Simpson, E A1 - Dazzi, F J1 - Vox Sang Y1 - 2004/07// VL - 87 Suppl 2 SN - 0042-9007 SP - 11 EP - 14 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15209868&query_hl=1 ER - TY - JFULL T1 - The introduction of TNF blockade in rheumatoid arthritis A1 - Maini, RN A1 - Feldmann, M J1 - ANN RHEUM DIS Y1 - 2004/07// VL - 63 SN - 0003-4967 SP - 13 EP - 13 ER - TY - JFULL T1 - Menage a trois of bacterial and viral pulmonary pathogens delivers coup de grace to the lung A1 - Hussell, T A1 - Williams, A J1 - CLIN EXP IMMUNOL Y1 - 2004/07// VL - 137 SN - 0009-9104 SP - 8 EP - 11 ER - TY - JFULL T1 - Patterns of autoimmunity in primary biliary cirrhosis patients and their families: a population-based cohort study. A1 - Watt, FE A1 - James, OF A1 - Jones, DE J1 - QJM Y1 - 2004/07// VL - 97 SN - 1460-2725 SP - 397 EP - 406 N2 - BACKGROUND: Primary biliary cirrhosis (PBC) is a chronic liver disease with autoimmune features but uncertain aetiology. Increased risk of PBC among relatives of patients may reflect common environmental factors, or inherited immunogenetic susceptibility. Associations between PBC and other autoimmune diseases have been reported, but their true extent and pattern is unknown. Aim: To examine the prevalence and association patterns of autoimmune disease in a representative group of PBC patients. DESIGN: Clinical cohort study. METHODS: We clinically assessed members of a geographically-based PBC patient cohort (n = 160) for the presence of additional autoimmune disease, using established specific diagnostic criteria. RESULTS:Some 53% of patients had at least one additional autoimmune condition, and 63% had serum autoantibodies other than AMA or ANA. AMA+ patients had a significantly lower prevalence of additional autoimmunity than AMA- patients (49% vs. 79%; p < 0.01). The greatest relative increase in disease prevalence was for scleroderma (8% of patients). Autoimmune disease was present in 14% of first-degree relatives. DISCUSSION: PBC patients and their families have a wide susceptibility to autoimmunity. This observation supports an autoimmune aetiology and suggests that the genetic basis of PBC is likely to be expressed, at least in part, through factors controlling immune tolerance in general. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15208427&query_hl=1 ER - TY - JFULL T1 - Anti-IL6 receptor therapy: Rationale and early results in rheumatoid arthritis A1 - Maini, RN J1 - ANN RHEUM DIS Y1 - 2004/07// VL - 63 SN - 0003-4967 SP - 25 EP - 26 ER - TY - JFULL T1 - Proteomic analysis of Leishmania mexicana differentiation. A1 - Nugent, PG A1 - Karsani, SA A1 - Wait, R A1 - Tempero, J A1 - Smith, DF J1 - Mol Biochem Parasitol Y1 - 2004/07// VL - 136 SN - 0166-6851 SP - 51 EP - 62 N2 - We have resolved the proteome of axenically differentiated Leishmania mexicana parasites by two-dimensional gel electrophoresis (2DE), employing optimised, robust and reproducible procedures, and visualised (by silver staining) approximately 2000 protein species in each of three developmental stages: procyclic promastigotes, metacyclic promastigotes and amastigotes. This analysis has used homogeneous populations of these parasite stages, characterised according to their morphology, protease and nuclease activity profiles and expression of stage-specific antigens. Following comparison of the whole proteome profiles between stages, 47 spots were found to be stage-specific, while a further 100 spots changed in intensity during differentiation. The majority of "unique" spots were expressed during the infective stages of parasite differentiation, metacyclic promastigotes and amastigotes. CapLC-QTOF mass spectrometry has allowed the identification of 47 protein species to date, including a number which are only detected in the amastigote stage. Proteins identified are members of eight functionally related groupings, some of which are implicated in infectivity and host-parasite interactions. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15138067&query_hl=1 ER - TY - JFULL T1 - Increased keratin content detected by proteomic analysis of exhaled breath condensate from healthy persons who smoke. A1 - Gianazza, E A1 - Allegra, L A1 - Bucchioni, E A1 - Eberini, I A1 - Puglisi, L A1 - Blasi, F A1 - Terzano, C A1 - Wait, R A1 - Sirtori, CR J1 - Am J Med Y1 - 2004/07/01/ VL - 117 SN - 0002-9343 SP - 51 EP - 54 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15210388&query_hl=1 ER - TY - JFULL T1 - Palmitoylation of MT1-MMP at cysteine 574 in the cytoplasmic tail facilitates its internalisation and modulate cell migration A1 - Anilkumer, N A1 - Uekita, T A1 - Couchman, J A1 - Seiki, M A1 - Nagase, H A1 - Itoh, Y J1 - BRIT J CANCER Y1 - 2004/07// VL - 91 SN - 0007-0920 SP - S72 EP - S72 ER - TY - JFULL T1 - Treatment with infliximab leads to decrease in joint erosion scores in early rheumatoid arthritis patients with existing erosions A1 - van der Heijde, D A1 - Westhovens, R A1 - Keystone, E A1 - Maini, RN A1 - Emery, P A1 - Shiff, M A1 - Kalden, JR A1 - Bathon, J A1 - Baker, D A1 - Patel, K A1 - Han, J A1 - Clair, EWS A1 - Smolen, J J1 - ANN RHEUM DIS Y1 - 2004/07// VL - 63 SN - 0003-4967 SP - 294 EP - 294 ER - TY - JFULL T1 - A proteomic analysis of changes in prothrombin and plasma proteins associated with the G20210A mutation. A1 - Gelfi, C A1 - Viganò, A A1 - Ripamonti, M A1 - Wait, R A1 - Begum, S A1 - Biguzzi, E A1 - Castaman, G A1 - Faioni, EM J1 - Proteomics Y1 - 2004/07// VL - 4 SN - 1615-9853 SP - 2151 EP - 2159 N2 - The G-->A mutation at position 20210 of the prothrombin gene, localized in the 3'-polyadenylation untranslated region of the mRNA, is a recognized genetic risk factor for venous thromboembolism. The mechanism by which this base change confers an increased risk of thrombosis compared to noncarriers is undefined. Studies on the mRNA suggest enhanced cleavage site recognition and a change in the location of the 3'-cleavage/polyadenylation reaction, but no defined model has been proposed. The present study, based on proteomic investigation by two-dimensional gel electrophoresis and electrospray ionization (ESI) tandem mass spectrometry (MS/MS) protein identification, suggests that the G20210A mutation is associated with increased glycosylation of prothrombin, which confers greater stability to the protein. Additionally, proteomic investigation of pooled plasma showed that expression levels of six spots, three of them identified by ESI MS/MS, were altered in subjects carrying the mutation, suggesting a possible cooperative effect in the thrombotic risk increment induced by the mutation. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15221775&query_hl=1 ER - TY - JFULL T1 - The role of TNF ininflammatory joint disease A1 - Feldmann, M A1 - Maini, RN J1 - ANN RHEUM DIS Y1 - 2004/07// VL - 63 SN - 0003-4967 SP - 12 EP - 13 ER - TY - JFULL T1 - Infliximab therapy leads to clinical and radiographic benefits in early rheumatoid arthritis patients receiving optimum methotrexate dose A1 - Bathon, J A1 - Devogelaer, J A1 - Kalden, JR A1 - Shiff, M A1 - Emery, P A1 - Maini, RN A1 - van der Heijde, D A1 - Keystone, E A1 - Baker, D A1 - Patel, K A1 - Han, J A1 - Smolen, J A1 - Clair, EWS J1 - ANN RHEUM DIS Y1 - 2004/07// VL - 63 SN - 0003-4967 SP - 293 EP - 293 ER - TY - JFULL T1 - Serum levels of IL-6 and TNF-alpha correlate with clinicopathological features and patient survival in patients with prostate cancer. A1 - Michalaki, V A1 - Syrigos, K A1 - Charles, P A1 - Waxman, J J1 - Br J Cancer Y1 - 2004/06/14/ VL - 90 SN - 0007-0920 SP - 2312 EP - 2316 N2 - Interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-alpha) are important multifunctional cytokines involved in tumour growth and metastasis. In this study, we have measured serial levels of serum IL-6 and TNF-alpha in prostate cancer patients. A total of 80 patients with carcinoma of the prostate and 38 controls were studied. Three patient groups, with small bulk localised, large volume localised and metastatic prostate cancer, were assessed. Serum IL-6 and TNF-alpha levels were measured and correlated with clinicopathological variables and patient survival. Serial changes in these cytokines were also assessed and related to disease progression in 40 patients with recurrent prostate cancer. Serum IL-6 levels in patients with metastatic disease (9.3+/-7.8 pg x ml(-1)) were higher than those in patients with localised disease (1.3+/-0.8 pg x ml(-1), P<0.001). Significantly elevated levels of TNF-alpha were found in metastatic disease (6.3+/-3.6 pg x ml(-1)) compared with localised disease (1.1+/-0.5 pg x ml(-1), P<0.001). The levels of both cytokines were directly correlated with the extent of the disease. Serial analysis in 40 patients with recurrent tumours showed that both cytokines became elevated at the point of prostate-specific antigen progression. In conclusion, these results suggest that IL-6 and TNF-alpha correlate with the extent of disease in patients with prostate cancer and may be monitored in conjunction with other disease markers. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15150588&query_hl=1 ER - TY - JFULL T1 - Tissue repair and the dynamics of the extracellular matrix. A1 - Midwood, KS A1 - Williams, LV A1 - Schwarzbauer, JE J1 - Int J Biochem Cell Biol Y1 - 2004/06// VL - 36 SN - 1357-2725 SP - 1031 EP - 1037 N2 - Repair of tissue after injury depends on the synthesis of a fibrous extracellular matrix to replace lost or damaged tissue. Newly deposited extracellular matrix is then re-modeled over time to emulate normal tissue. The extracellular matrix directs repair by regulating the behavior of the wide variety of cell types that are mobilized to the damaged area in order to rebuild the tissue. Acute inflammation, re-epithelialization, and contraction all depend on cell-extracellular matrix interactions and contribute to minimize infection and promote rapid wound closure. Matricellular proteins are up-regulated during wound healing where they modulate interactions between cells and the extracellular matrix to exert control over events that are essential for efficient tissue repair. Here, we discuss how the extracellular matrix changes during the stages of tissue repair, how matricellular proteins affect cell-extracellular matrix interactions, and how these proteins might be exploited for use therapeutically. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15094118&query_hl=1 ER - TY - JFULL T1 - Feasibility of thoracoscopic U-clip esophageal anastomosis: an alternative for esophageal atresia reconstruction. A1 - Tirabassi, MV A1 - Banever, GT A1 - Moriarty, KP A1 - Konefal, S A1 - Reiter, E A1 - Wait, R J1 - J Pediatr Surg Y1 - 2004/06// VL - 39 SN - 1531-5037 SP - 851 EP - 854 N2 - BACKGROUND: The authors propose that U-Clips can significantly decrease the technical difficulty of performing thoracoscopic esophageal reconstruction, thus, reducing operating time, the incidence of postoperative leak, and stricture rate. METHODS: After obtaining Institutional Animal Care and Use Committee approval, 3 4-kg female piglets underwent complete thoracoscopic esophageal transections. The esophagus was reconstructed thoracoscopically using S50 and S60 U-Clips over an 8F transanastomotic tube. Esophagrams were performed on postoperative day (POD) 7, 21, 44, and 77. RESULTS: Mean operating time was 57 minutes (45 to 75 min). Two of 3 piglets had no evidence of leak on POD 7 esophagrams. One animal had a small leak that resolved spontaneously on antibiotics. All 3 piglets tolerated a formula diet orally by POD 8. Over a 77-day survival period all 3 piglets had steady weight gain on an oral diet. CONCLUSIONS: U-Clips are a feasible alternative to sutures for esophageal reconstruction in thoracoscopic surgery. Further study is warranted to investigate the full potential of U-Clips in minimally invasive pediatric surgery. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15185211&query_hl=1 ER - TY - JFULL T1 - Control of human articular chondrocyte differentiation by reduced oxygen tension. A1 - Murphy, CL A1 - Polak, JM J1 - J Cell Physiol Y1 - 2004/06// VL - 199 SN - 0021-9541 SP - 451 EP - 459 N2 - Cell number is often a limiting factor in studies of chondrocyte physiology, particularly for human investigations. Chondrocytes can be readily proliferated in monolayer culture, however, differentiated phenotype is soon lost. We therefore endeavored to restore normal phenotype to human chondrocytes after serial passage in monolayer culture by manipulating cell morphology and oxygen tension towards the in vivo state. Third passage cells were encapsulated in alginate and exposed to either 20% or more physiologic 5% oxygen tensions. To assess cell phenotype, gene expression was measured using TaqMan real-time PCR. Encapsulated, primary chondrocytes cultured in 20% oxygen were used as a positive reference. Passaged human chondrocytes were fibroblastic in appearance and had lost normal phenotype as evidenced by a decrease in expression of collagen II, aggrecan, and sox9 genes of 66, 6, and 14 fold, respectively; with concomitant high expression of type I collagen (22 fold increase). A partial regaining of the differentiated phenotype was observed by encapsulation in 20% oxygen; however, even after 4 weeks, collagen II gene expression was not fully restored. Collagen II and aggrecan expression were increased, on average, 3 fold, in 5% oxygen tension compared to 20% cultures. Furthermore, matrix glycosaminoglycan (GAG) levels were significantly increased in reduced oxygen. In fact, after 4 weeks in 5% oxygen, encapsulated third passage cells had collagen II expression fully regained and aggrecan and sox9 levels actually exceeding primary cell levels in 20% oxygen. Our results show that the phenotype of serially passaged human articular chondrocytes is more fully restored by combining encapsulation with culture in more physiological levels of oxygen. Sox9, an essential transcription factor for chondrocyte differentiation is strongly implicated in this process since its expression was upregulated almost 27 fold. These findings have implications for the optimal conditions for the in vitro culture of chondrocytes. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15095292&query_hl=1 ER - TY - JFULL T1 - A monoclonal antibody to glucosylceramide inhibits the growth of Fonsecaea pedrosoi and enhances the antifungal action of mouse macrophages. A1 - Nimrichter, L A1 - Barreto-Bergter, E A1 - Mendonça-Filho, RR A1 - Kneipp, LF A1 - Mazzi, MT A1 - Salve, P A1 - Farias, SE A1 - Wait, R A1 - Alviano, CS A1 - Rodrigues, ML J1 - Microbes Infect Y1 - 2004/06// VL - 6 SN - 1286-4579 SP - 657 EP - 665 N2 - Fungal glucosylceramides (GlcCer) are conserved lipid components in a large variety of pathogenic and non-pathogenic fungal species, but their biological functions are still unclear. Recent studies demonstrate that GlcCer are immunologically active components inducing the production of antifungal antibodies. In this work, a major GlcCer was purified and characterized from mycelial forms of Fonsecaea pedrosoi, the most frequent causative agent of chromoblastomycosis. As determined by fast atom bombardment mass spectrometry (FAB-MS) analysis, the purified molecule was identified as the conserved structure N-2'-hydroxyhexadecanoyl-1-beta-D-glucopyranosyl-9-methyl-4,8-sphingadienine. A monoclonal antibody (Mab) against this structure was used in indirect immunofluorescence with the different morphological stages of F. pedrosoi. Only the surface of young dividing cells was recognized by the antibody. Treatment of F. pedrosoi conidia with the Mab against GlcCer resulted in a clear reduction in fungal growth. In addition, the Mab also enhanced phagocytosis and killing of F. pedrosoi by murine cells. These results suggest that, in F. pedrosoi, GlcCer seem to be cell wall components targeted by antifungal antibodies that directly inhibit fungal development and also enhance macrophage function, supporting the use of monoclonal antibodies to GlcCer as potential tools in antifungal immunotherapy. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15158773&query_hl=1 ER - TY - JFULL T1 - Flagellin promotes myeloid differentiation factor 88-dependent development of Th2-type response. A1 - Didierlaurent, A A1 - Ferrero, I A1 - Otten, LA A1 - Dubois, B A1 - Reinhardt, M A1 - Carlsen, H A1 - Blomhoff, R A1 - Akira, S A1 - Kraehenbuhl, JP A1 - Sirard, JC J1 - J Immunol Y1 - 2004/06/01/ VL - 172 SN - 0022-1767 SP - 6922 EP - 6930 N2 - Activation of dendritic cells (DC) by microbial products via Toll-like receptors (TLR) is instrumental in the induction of immunity. In particular, TLR signaling plays a major role in the instruction of Th1 responses. The development of Th2 responses has been proposed to be independent of the adapter molecule myeloid differentiation factor 88 (MyD88) involved in signal transduction by TLRs. In this study we show that flagellin, the bacterial stimulus for TLR5, drives MyD88-dependent Th2-type immunity in mice. Flagellin promotes the secretion of IL-4 and IL-13 by Ag-specific CD4(+) T cells as well as IgG1 responses. The Th2-biased responses are associated with the maturation of DCs, which are shown to express TLR5. Flagellin-mediated DC activation requires MyD88 and induces NF-kappaB-dependent transcription and the production of low levels of proinflammatory cytokines. In addition, the flagellin-specific response is characterized by the lack of secretion of the Th1-promoting cytokine IL-12 p70. In conclusion, this study suggests that flagellin and, more generally, TLR ligands can control Th2 responses in a MyD88-dependent manner. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15153511&query_hl=1 ER - TY - JFULL T1 - MT1-MMP: an enzyme with multidimensional regulation. A1 - Itoh, Y A1 - Seiki, M J1 - Trends Biochem Sci Y1 - 2004/06// VL - 29 SN - 0968-0004 SP - 285 EP - 289 N2 - The activity of membrane-type 1 matrix metalloproteinase (MT1-MMP) is a double-edged sword--it is crucial for both physiological processes and disease progression. MT1-MMP modifies various cellular functions and it is, sthus, regulated precisely as a proteinase and as a membrane protein. Recent studies have further revealed that the function of MT1-MMP is modified and regulated by O-glycosylation, interaction with CD44, internalization and recycling. Such multidimensional mechanisms enable MT1-MMP to be regulated spatially and temporally, and are essential for its proper functioning on the cell surface. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15276180&query_hl=1 ER - TY - JFULL T1 - Sjögren's syndrome. A1 - Venables, PJ J1 - Best Pract Res Clin Rheumatol Y1 - 2004/06// VL - 18 SN - 1521-6942 SP - 313 EP - 329 N2 - Sjögren's syndrome is an autoimmune disease characterized by inflammation of the exocrine glands, leading to impaired function. Here, I review the relatively short history of the syndrome and explain why it is frequently underdiagnosed, undertreated and under-researched. Attempts to provide classification criteria have culminated in the revised American-European Consensus Criteria, which provide a sound basis for both clinical management and research. The recognition that Sjögren's syndrome is a disease of considerable morbidity has led to a more aggressive approach to therapy ranging from topical therapies to systemic treatment with secretagogues such as pilocarpine and cemiveline, and immunomodulatory drugs such as hydroxychloroquine and interferon-alpha. The central role of the glandular epithelial cell is identified as the key to understanding the pathogenesis of the disease. Hypofunction rather than destruction of these cells is now regarded as the main mechanism of secretory failure in Sjögren's syndrome. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15158743&query_hl=1 ER - TY - JFULL T1 - Closing remarks A1 - Maini, RN J1 - ARTHRITIS RES THER Y1 - 2004/06// VL - 6 SN - 1478-6362 SP - S44 EP - S44 ER - TY - JFULL T1 - Manipulation of immunity to and pathology of respiratory infections. A1 - Snelgrove, R A1 - Williams, A A1 - Thorpe, C A1 - Hussell, T J1 - Expert Rev Anti Infect Ther Y1 - 2004/06// VL - 2 SN - 1478-7210 SP - 413 EP - 426 N2 - Respiratory infections are the third leading cause of death worldwide and are a priority for vaccine development. Immune defence mechanisms are critical in recovery from most respiratory infections but the role of the immune system in causing bystander lung injury is not as well understood, and will be the focus of this review. Immune-mediated injury results from physical occlusion of the airways or the ensuing 'cytokine storm', which may spill over into the systemic circulation and cause devastating consequences. Respiratory pathogens employ numerous strategies to avoid detection by the immune system. One of these, the alteration of key surface determinants, makes the design of rational vaccines problematic. In the following review the immune compartments responsible for clinical lung disease are discussed, and current and novel strategies to reduce their potency are overviewed. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15482206&query_hl=1 ER - TY - JFULL T1 - Increased in vivo transcription of an IL-8 haplotype associated with respiratory syncytial virus disease-susceptibility. A1 - Hacking, D A1 - Knight, JC A1 - Rockett, K A1 - Brown, H A1 - Frampton, J A1 - Kwiatkowski, DP A1 - Hull, J A1 - Udalova, IA J1 - Genes Immun Y1 - 2004/06// VL - 5 SN - 1466-4879 SP - 274 EP - 282 N2 - Interleukin-8 (IL-8) has been implicated in the pathogenesis of RSV-induced bronchiolitis. Previously, we have described an association between bronchiolitis disease severity and a specific IL-8 haplotype comprising six single-nucleotide polymorphisms (SNPs) (-251A/+396G/+781T/+1238delA/+1633T/+2767T, haplotype 2). Here we investigated the functional basis for this association by measuring haplotype-specific transcription in vivo in human primary cells. We found a significant increase in transcript level derived from the IL-8 haplotype 2 relative to the mirror haplotype 1 (-251T/+396T/+781C/+1238insA/+1633C/+2767A) in respiratory epithelial cells but not in lymphocytes. A promoter polymorphism, -251A, present on the high producer haplotype, had no significant affect on the allele-specific level of transcription when analyzed in reporter gene experiments in human respiratory epithelial A549 cells. We proceeded to systematically screen for allele-specific protein-DNA binding in this functional haplotype, which revealed significant differential binding at the +781T/C polymorphism. C/EBP beta was identified as being part of a transcription factor binding complex that preferentially bound in the presence of the +781 T allele. These results suggest that the mechanism for disease susceptibility to RSV-induced bronchiolitis may occur through a haplotype-specific increase in IL-8 transcription, which may be mediated by functional polymorphisms within that haplotype. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15085176&query_hl=1 ER - TY - JFULL T1 - Conventional protein kinase C and atypical protein kinase Czeta differentially regulate macrophage production of tumour necrosis factor-alpha and interleukin-10. A1 - Foey, AD A1 - Brennan, FM J1 - Immunology Y1 - 2004/05// VL - 112 SN - 0019-2805 SP - 44 EP - 53 N2 - In chronic inflammatory diseases such as rheumatoid arthritis, joint macrophages/monocytes are the major source of pro- and anti-inflammatory cytokines. Little is understood regarding the signalling pathways which determine the production of the pro-inflammatory cytokine, tumour necrosis factor-alpha (TNF-alpha) and the anti-inflammatory cytokine, interleukin-10 (IL-10). Two pathways integral to macrophage function are the protein kinase C (PKC)- and the cAMP-dependent pathways. In this report, we have investigated the involvement of PKC and cAMP in the production of TNF-alpha and IL-10 by peripheral blood monocyte-derived macrophages. The utilization of the PKC inhibitors Go6983, Go6976 and RO-32-0432 demonstrated a role for conventional PKCs (alpha and beta) in the production of TNF-alpha in response to stimulation by lipopolysaccharide and phorbol 12-myristate 13-acetate (PMA)/ionomycin. PKC stimulation resulted in the downstream activation of the p42/44 mitogen-activated protein kinase (MAPK) pathway which differentially regulates TNF-alpha and IL-10. The addition of cAMP however, suppressed activation of this MAPK and TNF-alpha production. Cyclic-AMP augmented IL-10 production and cAMP response element binding protein activation upon stimulation by PMA/ionomycin. In addition, cAMP activated PKCzeta; inhibition of which, by a dominant negative adenovirus construct, selectively suppressed IL-10 production. These observations suggest that pro-inflammatory and anti-inflammatory cytokines are differentially regulated by PKC isoforms; TNF-alpha being dependent on conventional PKCs (alpha and beta) whereas IL-10 is regulated by the cAMP-regulated atypical PKCzeta. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15096183&query_hl=1 ER - TY - JFULL T1 - Disease modifying treatment and elective surgery in rheumatoid arthritis: the need for more data A1 - Jain, A A1 - Maini, R A1 - Nanchahal, J J1 - ANN RHEUM DIS Y1 - 2004/05/01/ VL - 63 SN - 0003-4967 SP - 602 EP - 603 ER - TY - JFULL T1 - The Role Of The NF kappa B Pathway In Atherosclerosis A1 - Monaco C A1 - Grosjean J A1 - Paleolog E J1 - International Atherosclerosis Society Web-Site Y1 - 2004/05// UR - http://www.athero.org/commentaries/comm303.pdf ER - TY - JFULL T1 - Conservation and loss of the ERV3 open reading frame in primates. A1 - Hervé, CA A1 - Forrest, G A1 - Löwer, R A1 - Griffiths, DJ A1 - Venables, PJ J1 - Genomics Y1 - 2004/05// VL - 83 SN - 0888-7543 SP - 940 EP - 943 N2 - The human endogenous retrovirus ERV3 possesses an open reading frame for a truncated envelope, which is expressed as mRNA and protein. Here we examine the env sequence in primates for evidence of evolutionary conservation. ERV3 sequences were amplified by PCR from genomic DNA of great ape and Old World primates but not from New World primates or gorilla, suggesting an integration event more than 30 million years ago with a subsequent loss in one species. In the chimpanzee, the protein sequence of Env is 98.18% identical to that of human. In other species the identity falls (93.71% in rhesus macaque) in proportion to the separation from the human lineage. Start and stop codons and domains of functional significance in the envelope protein are conserved. The evolutionary conservation of the ERV3 envelope suggests a beneficial function, though the loss from gorilla shows that it is not essential for survival or reproduction. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15081124&query_hl=1 ER - TY - JFULL T1 - Shear stress-induced shedding of soluble intercellular adhesion molecule-1 from saphenous vein endothelium. A1 - Sultan, S A1 - Gosling, M A1 - Nagase, H A1 - Powell, JT J1 - FEBS Lett Y1 - 2004/04/23/ VL - 564 SN - 0014-5793 SP - 161 EP - 165 N2 - Within 6 h, shear stress upregulated intercellular adhesion molecule-1 (ICAM-1) (two- to four-fold, P<0.001) and induced matrix metalloproteinase-2 (MMP-2) in cultured human saphenous vein endothelial cells. By 8 h endothelial ICAM-1 levels returned to baseline, with concomitant increase in soluble ICAM-1 (sICAM-1) (P<0.001) and MMP-9 had been induced. Inclusion of a hydroxamate metalloproteinase inhibitor partially reversed the effects on ICAM-1 and sICAM-1 at 8 h, whereas TIMP-1, -2 or -3 had no effect. MMP-9, but not MMP-2, co-immunoprecipitated with ICAM-1. sICAM-1 was processed distal to Arg441, indicating that MMP-9, docking to ICAM-1, contributes to sICAM-1 shedding and attenuation of the shear stress-induced upregulation of ICAM-1. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15094060&query_hl=1 ER - TY - JFULL T1 - Androgens target prohibitin to regulate proliferation of prostate cancer cells. A1 - Gamble, SC A1 - Odontiadis, M A1 - Waxman, J A1 - Westbrook, JA A1 - Dunn, MJ A1 - Wait, R A1 - Lam, EW A1 - Bevan, CL J1 - Oncogene Y1 - 2004/04/15/ VL - 23 SN - 0950-9232 SP - 2996 EP - 3004 N2 - Proteins involved in the growth response of prostate cancer cells to androgen were investigated by comparing the proteomes of LNCaP cells treated with vehicle or androgen. Whole-cell lysates were separated by two-dimensional PAGE, and HPLC-MS/MS was used to identify androgen-regulated proteins. Prohibitin, a protein with cell-cycle regulatory activity, was shown to be downregulated by 50% following androgen stimulation. Western blot and reverse transcription-PCR experiments confirmed the result and showed that regulation occurs at the level of transcription. To determine the importance of prohibitin in androgen-stimulated growth, we used transient transfection to overexpress the protein and RNA interference to knock down the protein. Subsequent FACS analysis showed that cells with reduced levels of prohibitin showed a slight but reproducible increase in the percentage of population in cell cycle, while cells with increased prohibitin levels showed a clear reduction in the percentage entering cell cycle, following dihydrotestosterone stimulation, when compared to untransfected controls. Confocal microscopy showed localization of prohibitin in the nucleus as well as the mitochondria of LNCaP cells. It therefore seems that the regulation of prohibitin is a vital part of the cellular growth response to androgen stimulation in LNCaPs and prohibitin may have a nuclear regulatory role in cell-cycle progression. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=14968116&query_hl=1 ER - TY - JFULL T1 - Canonical pathway of nuclear factor kappa B activation selectively regulates proinflammatory and prothrombotic responses in human atherosclerosis. A1 - Monaco, C A1 - Andreakos, E A1 - Kiriakidis, S A1 - Mauri, C A1 - Bicknell, C A1 - Foxwell, B A1 - Cheshire, N A1 - Paleolog, E A1 - Feldmann, M J1 - Proc Natl Acad Sci U S A Y1 - 2004/04/13/ VL - 101 SN - 0027-8424 SP - 5634 EP - 5639 N2 - Nuclear factor kappa B (NF-kappa B) activation has been observed in human atherosclerotic plaques and is enhanced in unstable coronary plaques, but whether such activation has a protective or pathophysiological role remains to be determined. We addressed this question by developing a short-term culture system of cells isolated from human atherosclerotic tissue, allowing efficient gene transfer to directly investigate signaling pathways in human atherosclerosis. We found that NF-kappa B is activated in these cells and that this activity involves p65, p50, and c-Rel but not p52 or RelB. This NF-kappa B activation can be blocked by overexpression of I kappa B alpha or dominant-negative I kappa B kinase (IKK)-2 but not dominant-negative IKK-1 or NF-kappa B-inducing kinase, resulting in selective inhibition of inflammatory cytokines (tumor necrosis factor alpha, IL-6, and IL-8), tissue factor, and matrix metalloproteinases without affecting the antiinflammatory cytokine IL-10 or tissue inhibitor of matrix metalloproteinases. Our results demonstrate that the canonical pathway of NF-kappa B activation that involves p65, p50, c-Rel, and IKK-2 is activated in human atherosclerosis and results in selective up-regulation of major proinflammatory and prothrombotic mediators of the disease. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15064395&query_hl=1 ER - TY - JFULL T1 - Identification and characterization of 4-[[4-(2-butynyloxy) phenyl]sulfonyl]-N-hydroxy-2,2-dimethyl-(3S)-thiomorpholinecarboxamide (TMI-1), a novel dual tumor necrosis factor-alpha-converting enzyme/matrix metalloprotease inhibitor for the treatment of rheumatoid arthritis A1 - Zhang, YH A1 - Xu, J A1 - Levin, J A1 - Hegen, M A1 - Li, GD A1 - Robertshaw, H A1 - Brennan, F A1 - Cummons, T A1 - Clarke, D A1 - Vansell, N A1 - Nickerson-Nutter, C A1 - Barone, D A1 - Mohler, K A1 - Black, R A1 - Skotnicki, J A1 - Gibbons, J A1 - Feldmann, M A1 - Frost, P A1 - Larsen, G A1 - Lin, LL J1 - J PHARMACOL EXP THER Y1 - 2004/04/01/ VL - 309 SN - 0022-3565 SP - 348 EP - 355 N2 - Tumor necrosis factor (TNF)-alpha is a well validated therapeutic target for the treatment of rheumatoid arthritis. TNF-alpha is initially synthesized as a 26-kDa membrane-bound form (pro-TNF) that is cleaved by a Zn-metalloprotease named TNF-alpha-converting enzyme (TACE) to generate the 17-kDa, soluble, mature TNF-alpha. TACE inhibitors that prevent the secretion of soluble TNF-alpha may be effective in treating rheumatoid arthritis ( RA) patients. Using a structure-based design approach, we have identified a novel dual TACE/matrix metalloprotease (MMP) inhibitor 4-[[ 4( 2-butynyloxy) phenyl] sulfonyl]-N-hydroxy-2,2-dimethyl-(3S)thiomorpholinecarboxamide( TMI-1). This molecule inhibits TACE and several MMPs with nanomolar IC50 values in vitro. In cell-based assays such as monocyte cell lines, human primary monocytes, and human whole blood, it inhibits lipopolysaccharide LPS)-induced TNF-alpha secretion at submicromolar concentrations, whereas there is no effect on the TNF-alpha mRNA level as judged by RNase protection assay. The inhibition of LPS-induced TNF-alpha secretion is selective because TMI-1 has no effect on the secretion of other proinflammatory cytokines such as interleukin (IL)-1beta, IL-6, and IL-8. Importantly, TMI-1 potently inhibits TNF-alpha secretion by human synovium tissue explants of RA patients. In vivo, TMI-1 is highly effective in reducing clinical severity scores in mouse prophylactic collagen-induced arthritis ( CIA) at 5, 10, and 20 mg/kg p.o. b.i.d. and therapeutic CIA model at 100 mg/kg p.o. b.i.d. In summary, TMI-1, a dual TACE/MMP inhibitor, represents a unique class of orally bioavailable small molecule TNF inhibitors that may be effective and beneficial for treating RA. ER - TY - JFULL T1 - Comparison of ultrasonographic assessment of synovitis and joint vascularity with radiographic evaluation in a randomized, placebo-controlled study of infliximab therapy in early rheumatoid arthritis. A1 - Taylor, PC A1 - Steuer, A A1 - Gruber, J A1 - Cosgrove, DO A1 - Blomley, MJ A1 - Marsters, PA A1 - Wagner, CL A1 - McClinton, C A1 - Maini, RN J1 - Arthritis Rheum Y1 - 2004/04// VL - 50 SN - 0004-3591 SP - 1107 EP - 1116 N2 - OBJECTIVE: To investigate sensitive ultrasonographic imaging methods for detection of synovial thickness and vascularity to discriminate between patients with early rheumatoid arthritis (RA) receiving infliximab + methotrexate (MTX) versus placebo + MTX over 18 weeks, and to compare the relationship between synovial thickening and vascularity at baseline and radiologic damage to joints of the hands and feet at 54 weeks. METHODS: Patients with early RA (duration <3 years) receiving stable dosages of MTX were randomly assigned to receive blinded infusions of 5 mg/kg infliximab (n = 12) or placebo (n = 12) at weeks 0, 2, 6, and then every 8 weeks until week 46. At baseline and week 18, clinical assessments were performed, and metacarpophalangeal joints were assessed by high-frequency ultrasonography and power Doppler ultrasonography measurements. Radiographs of the hands and feet taken at baseline and at 54 weeks were evaluated using the van der Heijde modification of the Sharp method (vdH-Sharp score). RESULTS: Using changes in the total vdH-Sharp score over 54 weeks and changes in synovial thickening and joint vascularity at 18 weeks, we were able to distinguish those patients receiving infusions of infliximab + MTX from those receiving placebo + MTX. Sonographic measurements of synovial thickening and vascularity at baseline in the placebo + MTX group demonstrated clear relationships with the magnitude of radiologic joint damage at week 54. Infliximab + MTX treatment abolished these relationships. CONCLUSION: The delay or reversal of inflammatory and joint-destructive mechanisms in patients with early RA was already apparent following 18 weeks of treatment with infliximab + MTX and was reflected in radiologic changes at 54 weeks. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15077292&query_hl=1 ER - TY - JFULL T1 - Historical review: cytokines as therapeutics and targets of therapeutics A1 - Vilcek J A1 - Feldmann M J1 - Trends in Pharmacological Sciences Y1 - 2004/04// IS - 4 VL - 25 SP - 201 EP - 219 ER - TY - JFULL T1 - An unusual case of ankle arthropathy. A1 - Abraham, S A1 - Cope, A J1 - Ann Rheum Dis Y1 - 2004/04// VL - 63 SN - 0003-4967 SP - 460 EP - 461 L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15020346&query_hl=1 ER - TY - JFULL T1 - P-selectin glycoprotein ligand 1 therapy ameliorates established collagen-induced arthritis in DBA/1 mice partly through the suppression of tumour necrosis factor. A1 - Sumariwalla, PF A1 - Malfait, AM A1 - Feldmann, M J1 - Clin Exp Immunol Y1 - 2004/04// VL - 136 SN - 0009-9104 SP - 67 EP - 75 N2 - We investigated the therapeutic potential of P-selectin glycoprotein ligand (PSGL)-1 in established collagen-induced arthritis (CIA) in DBA/1 mice. PSGL-1 is the high-affinity specific ligand for P-selectin and is thus important in cell recruitment to inflammatory sites. I-316 PSGL-1 or rPSGL-1Ig fusion protein were administered to mice after the onset of clinical arthritis for 10 days, and the effect of treatment on both clinical and histopathological progression of disease was studied. It was found that both PSGL-1 biologicals effectively suppressed progression of clinical arthritis, and this was accompanied by protection against damage of joint tissues. We sought to investigate a mechanism underlying the effect of rPSGL-1Ig on the reduction of clinical arthritis. Blockade of PSGL-1/P-selectin interaction blocks recruitment of leucocytes, thus we observed a notable reduction in viable cell numbers of synoviocytes from rPSGL-1Ig treated mice. In view of this finding we suspected an effect of treatment on the production of pro-inflammatory mediators such as bioactive tumour necrosis factor-alpha (TNF) in synovial membrane ex vivo cell cultures. Production of TNF was reduced in arthritic mice that had been treated with rPSGL-1Ig. To further investigate the mechanism of rPSGL-1Ig, we explored the possibility that PSGL-1 might also have a direct signalling effect on TNF release from inflammatory cells. Thus synoviocyte cultures from arthritic mice were incubated with rPSGL-1Ig. A significant reduction in the spontaneous bioactive TNF release from these cultures was noted. We therefore confirmed these surprising findings using cultures of a mouse macrophage like cell line RAW 264.7, stimulated by LPS. Our results indicate that both forms of PSGL-1 have significant therapeutic effects in CIA murine model of RA. The mechanism of action involves reduced cellularity of synovium as anticipated, along with a reduction in TNF production from inflammatory cells in the synovium. The latter mechanism needs further mechanistic analysis. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15030516&query_hl=1 ER - TY - JFULL T1 - Sustained improvement over two years in physical function, structural damage, and signs and symptoms among patients with rheumatoid arthritis treated with infliximab and methotrexate. A1 - Maini, RN A1 - Breedveld, FC A1 - Kalden, JR A1 - Smolen, JS A1 - Furst, D A1 - Weisman, MH A1 - St Clair, EW A1 - Keenan, GF A1 - van der Heijde, D A1 - Marsters, PA A1 - Lipsky, PE A1 - Anti-Tumor Necrosis Factor Trial in Rheumatoid Arthritis with Concomitant Therapy Study Group J1 - Arthritis Rheum Y1 - 2004/04// VL - 50 SN - 0004-3591 SP - 1051 EP - 1065 N2 - OBJECTIVE: To evaluate the efficacy and safety of repeated administration of infliximab plus methotrexate (MTX) over a 2-year period in patients with rheumatoid arthritis (RA) who previously experienced an incomplete response to MTX. METHODS: Four hundred twenty-eight patients were randomly assigned to receive MTX plus placebo or infliximab at a dose of 3 or 10 mg/kg plus MTX for 54 weeks, with an additional year of followup. The protocol was later amended to allow for continued treatment during the second year. Of 259 patients who entered the second year of treatment, 216 continued to receive infliximab plus MTX for 102 weeks. Ninety-four of these 259 patients experienced a gap in therapy of >8 weeks before continuing therapy. Infusions were administered at weeks 0, 2, and 6, followed by treatment every 4 weeks or every 8 weeks (alternating with placebo infusions in the interim 4-week visits) at a dose of 3 or 10 mg/kg for a total of 102 weeks (including the gap in therapy). For safety and efficacy assessments, data on the patients who were randomized to receive treatment, irrespective of whether treatment was administered for 102 weeks, were evaluated using all actual observations available. The efficacy measures included the Health Assessment Questionnaire (HAQ) (physical function), Short Form 36 health survey (SF-36) (health-related quality of life), total radiographic scores (structural damage), and the American College of Rheumatology 20% improvement criteria (ACR20) (signs and symptoms). RESULTS: The infliximab plus MTX regimens resulted in significantly greater improvement in HAQ scores (P < or = 0.006) and SF-36 physical component summary scores (P < or = 0.011) compared with the MTX-only group. There also was stability in the SF-36 mental component summary score among patients who received the infliximab plus MTX regimens. Median changes from baseline to week 102 in the total radiographic score were 4.25 for patients who received the MTX-only regimen and 0.50 for patients who received the infliximab plus MTX regimen. The proportion of patients achieving an ACR20 response at week 102 varied from 40% to 48% for the infliximab plus MTX groups compared with 16% for the MTX-only group. CONCLUSION: Throughout 102 weeks of therapy, infliximab plus MTX provided significant, clinically relevant improvement in physical function and quality of life, accompanied by inhibition of progressive joint damage and sustained improvement in the signs and symptoms of RA among patients who previously had an incomplete response to MTX alone. L1 - http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15077287&query_hl=1 ER - TY - JFULL T1 - Historical review: Cytokines as therapeutics and targets of therapeutics A1 - Vilcek, J A1 - Feldmann, M J1 - TRENDS PHARMACOL SCI Y1 - 2004/04// VL - 25 SN - 0165-6147 SP - 201 EP - 209 N2 - Cytokine research has spawned the introduction of new therapies that have revolutionized the treatment of many important diseases. These therapeutic advances have resulted from two very different strategies. The first therapeutic strategy embodies the administration of purified, recombinant cytokines. The second relies on the administration of therapeutics that inhibit the harmful effects of upregulated, endogenous cytokines. Examples of successful cytokine therapeutics include hematopoietic growth factors (colony stimulating factors) and interferons. Prime examples of cytokine antagonists that have profoundly altered the treatment of some inflammatory disorders are agents that inhibit the effects of tumor necrosis factor (TNF). In this article, we highlight some of the studies that have been responsible for the introduction of cytokine and anti-cytokine therapies, with emphasis on the development of interferons and anti-TNIF agents. ER - TY - JFULL T1 - Membrane-type 1 matrix metalloproteinase cytoplasmic tail-binding protein-1 is a new member of the Cupin superfamily. A possible multifunctional protein acting as an invasion suppressor down-regulated in tumors. A1 - Uekita, T A1 - Gotoh, I A1 - Kinoshita, T A1 - Itoh, Y A1 - Sato, H A1 - Shiomi, T A1 - Okada, Y A1 - Seiki, M J1 - J Biol Chem Y1 - 2004/03/26/ VL - 279 SN - 0021-9258 SP - 12734 EP - 12743 N2 - Membrane-type 1 matrix metalloproteinase (MT1-MMP/MMP-14) is an enzyme that promotes tumor cell invasion in tissues. Although the proteolytic activity of MT1-MMP is indispensable for invasion, it is also regulated by functions of the cytoplasmic tail. In this study we obtained a new human gene whose product binds to the tail sequence in yeast. The product, MTCBP-1, is a 19-kDa protein that belongs to the newly proposed Cupin superfamily composed of proteins with diverse functions. MTCBP-1 expressed in cells formed a complex with MT1-MMP and co-localized at the membrane. It was also detected in both the cytoplasm and nucleus, where MT1-MMP does not exist. In human tumor cell lines MTCBP-1 expression was significantly low compared with non-transformed fibroblasts, and enforced expression of MTCBP-1 inhibited the activity of MT1-MMP in promoting cell migration and invasion. MTCBP-1 showed significant homology to the bacterial aci-reductone dioxygenase, which is an enzyme in methionine metabolism. The C-terminal part of