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Session III : Protein kinase cascades as therapeutic targets Poster III, 52 MAPKinase gene expression – as determined by microarray analysis - distinguishes uncomplicated from complicated reconstitution after major surgical trauma E. Marion Schneider1, Manfred Weiss2, Weidong Du1, Fabian Kriebel1, Gerhard Leder3, Klaus Buttenschön3, Ulrich C. Liener3, Uwe Bernd Brückner3 fabiankriebel@web.de 1Sektion Experimentelle Anaesthesiologie; 2Abteilung Klinische Anaesthesiologie, 3Abteilung Chirurgie, Universitaetsklinikum Ulm, Steinhoevelstrasse 9; 89075 Ulm, Germany Objective: A prospective pilot study in patients with major surgical trauma at an university hospital was set up to identify signaling pathways by microarray expression analysis, which may be distinct in individual patients and may be indicative for complicated versus uneventful reconstitution post trauma. Methods: RNA was prepared from peripheral blood drawn into PAXgene tubes from three patients before and exactly 24h after trauma. These RNA were then transcribed into cDNA, Cy3- or Cy5-labeled, and hybridized onto a cDNA microchip, red vs. green fluorescence was quantified using a microarray scanner. Patient (patients before and post trauma) versus control (healthy donors) blood cell gene expressions were compared using fluorescence intensities and appropriate normalizations. Results: In addition to a number of rather non-specific stress response genes, activated in all patients, we found a remarkable number of differences in gene expression patterns of individual patients. Some of the differing genes were associated with uncomplicated convalescence such as up-regulation of both the ERK5 pathway (MAPK7, mitogen-activated protein kinase-7) and transcription factors which stimulate hematopoiesis and tissue remodeling (MEF2, myocyte enhancer factor-2, BMP-2, bone morphogenic protein-2, TNFRSF11a (TNF-R superfamily, syn. RANK), and RUNX-1, runt related transcription factor-1). Chemokine genes active in stem cell recruitment from the bone marrow as well as dendritic cell and NK maturation (SCYA14 (HCC-1, hemofiltrate cc chemokine )), were increased as well as activators of the lymphoid compartment (TNFRSF7 (CD27), CD3zeta and perforin (PRF1)). In contrast, all these transcripts were down-regulated in complicated reconstitution and later development of septic shock. Moreover, p38 kinase (MAPK14), S100 molecules and members of the lipoxygenase pathway were associated with a more eventful outcome. In none of the patients, we found c-Jun-stress activated factor (JNK) pathway to be markedly activated, also p105 (Rel) or the “NFkB inducible kinase” (NIK, MAP3K14) were not found to be up-regulated post surgical trauma. Conclusions: Microarray expression studies are a promising tool for screening and then selecting differentially regulated genes in favorable as compared to complicated reconstitution post trauma. Gene up-regulation patterns may emphasize previously unrecognized molecules and signaling pathways encouraging the appropriate protein and functional assays in an individual patient but also in larger patient cohorts. - 238 -
Session III : Protein kinase cascades as therapeutic targets Poster III, 53 Modulation of MPF and MAPK pathways activities by physiological pHi changes. Chantal Sellier1, Anne F Antoine1, Stéphane Flament2, Arlette Lescuyer1, Jean-Pierre Vilain1 and Jean-François Bodart1. 1 - Laboratoire de Régulation des signaux de division, EA 1033, IFR 118, UTSL, SN3, F- 59655 Villeneuve d’Ascq cedex, France. E-mail : chantal_sellier@yahoo.fr ; Anne- Frederique.Antoine@univ-lille1.fr ; Arlette.Lescuyer@univ-lille1.fr ; Jean- Pierre.Vilain@univ-lille1.fr ; Jean-François.Bodart@univ-lille1.fr 2 - Faculté des Sciences, EA 3442, UHP, Bvd des Aiguillettes, BP 239, F-54506 Vandoeuvre Lès Nancy cedex, France. E-mail : stephane.flament@scbiol.uhp-nancy.fr Intracellular pH (pHi) plays a key role in mitogenic signal transduction, development and maintenance of transformed phenotypes. Nevertheless, cell cycle mechanisms sensitive to pHi remain to be determined. Analogous to G2/M transition, Xenopus oocyte maturation is characterized by germinal vesicle breakdown (GVBD), MPF (Cyclin B/Cdk1) and Mos- MAPK pathways activation. These events are accompanied by a transient alkalization. We determined that an acidifying compound, NH4Cl, delayed progesterone-induced GVBD in a dose-dependent manner. Cyclin B phosphorylation, Cdk1 Y-15 dephosphorylation as well as p39Mos accumulation, MAPK and p90Rsk phosphorylations induced by progesterone were delayed by acidification. The delay induced by NH4Cl was prevented by injection of MOPS buffer pH 7.7. In contrast, alkalyzing treatment such as Tris buffer pH 9 injections, accelerated GVBD, MPF and MAPK activation. Strickingly, we observed that NH4Cl strongly inhibited thiophosphorylated active MAPK-induced GVBD and MPF activation while it had no or little effect on the MPF autoamplification loop and GVBD triggered by egg cytoplasm injection. Nevertheless, Tris pH 9 did not have any effects on egg cytoplasm- or active MAPK-induced GVBD. Thus, dynamic of early events driving MAPK and MPF activation induced by progesterone may be negatively or positively regulated by pHi changes whereas only MAPK-induced GVBD pathway was inhibited by acidification. Finally, using electrophysiological approch, we observed that NH4Cl induced an activation of Ca2+activated Cl current suggesting an elevation of intracellular Ca2+ concentration. Therefore, effects of pHi on early meiosis dynamic might be related to subsequent change in Ca2+ homeostasis. - 239 -
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Table of content PREFACE ..........
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Preface In 1998, we organized the f
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Acknowledgments This meeting has be
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Group B (15h00 - 17h00) Session V.
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- 11 - Exhibition
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Visit our Expo (in alphabetical ord
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Thursday January 26th, 2006 (Early
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Thursday January 26th, 2006 (Evenin
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Regulation of inflammation and gluc
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Saturday January 28th, 2006 (Early
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Oral presentations (by alphabetical
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4: : Lottin S, Vercoutter-Edouart A
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the B-Raf/mitogen-activated/extrace
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Albertini MC, Accorsi A, Citterio B
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AP-1-dependent gene expression in s
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Jaks and cytokine receptors - an in
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The Role of Met-Gab1 Signalling in
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The mammalian tumor suppressor Scri
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SIGNAL TRANSDUCTION BY STRESS-ACTIV
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Title to be announced Caroline Dive
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Mechanisms of DNA methylation in ma
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Blocking the "4 Integrin-Paxillin I
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Regulation of NF-kB-dependent trans
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The immunomodulator AS101 induces g
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Signaling pathways leading to the a
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Boute, N., Jockers, R. and Issad, T
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Jespsen JS, Sorensen MD and Wengel
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8. Nishikawa, H., Kawai, K., Tanaka
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9) Proteome analysis of rat pancrea
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MicroRNA is an anti-apoptotic facto
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Resveratrol inhibits phorbol ester-
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Cross-talk between angiotensin II a
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Multiple role of histamine H 1 rece
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PI3K Targeting by the Beta-GBP Cyto
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D.P. Chopra, R.E. Menard, J. Janusz
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[2] Meijer, L., Flajolet, M. and Gr
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[4] Niemand, C., Nimmesgern, A., Ha
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Oncogenic signaling by mutant p53 M
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activity of the Peroxisome Prolifer
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Restauration of SHIP-1 activity in
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Hypoxia Signaling. Impact on Tumor
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5. Sokolov EI, Zykov KA, Pukhalsky
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HOW ANTITOXIC AND ANTICANCER AGENTS
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Listeria monocytogenes induced Rac1
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Epigenetic Regulation of Stem Cell
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Survey on in vitro cell death induc
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Distinct roles of IRF-3 and IRF-7 i
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Redox-Sensitive Transcription Facto
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Epigenic control of MHC2TA transcri
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Attenuation of MSK1-driven NF-kB ge
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Talking to chromatin: Silencers Sil
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Ubiquitin ligase Smurf1 controls os
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Workshop presentations (by alphabet
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Ambion MicroRNAs (miRNAs) are small
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Organizer: BIOBASE GmbH Date: Janua
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Reliable and efficient gene Knock-d
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Exploring ubiquitin signaling with
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Poster presentations Poster are cla
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Session I : protein domains Poster
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Session II. Receptor signaling and
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Session II : Receptor signaling and
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Session III. Protein kinase cascade
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Session III : Protein kinase cascad
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VIII. Inflammation specific signali
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Session X : Cell death in cancer Po
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Session XVI : Chemopreventive agent
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Dr. Nassera Aouali L-1526 Luxembour
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