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46 results on '"Service de Génomique Fonctionnelle"'

1. Immune thrombocytopenia and pregnancy: an exposed/nonexposed cohort study

Author

Stéphanie Guillet, Valentine Loustau, Emmanuelle Boutin, Anissa Zarour, Thibault Comont, Odile Souchaud-Debouverie, Nathalie Costedoat Chalumeau, Brigitte Pan-Petesch, Delphine Gobert, Stéphane Cheze, Jean Francois Viallard, Anne-Sophie Morin, Gaetan Sauvetre, Manuel Cliquennois, Bruno Royer, Agathe Masseau, Louis Terriou, Claire Fieschi, Olivier Lambotte, Stéphane Girault, Bertrand Lioger, Sylvain Audia, Karim Sacre, Jean Christophe Lega, Vincent Langlois, Alexandra Benachi, Corentin Orvain, Alain Devidas, Sebastien Humbert, Nicolas Gambier, Marc Ruivard, Virginie Zarrouk, Mikael Ebbo, Lise Willems, Lauriane Segaux, Matthieu Mahevas, Bassam Haddad, Marc Michel, Florence Canoui-Poitrine, Bertrand Godeau, Service de médecine interne [Mondor], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Etablissement Français du Sang [Île-de-France Mondor], IMRB - 'Transfusion et Maladies du Globule Rouge' [Créteil] (U955 Inserm - UPEC), Institut Mondor de Recherche Biomédicale (IMRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Laboratoire d'Excellence : Biogenèse et pathologies du globule rouge (Labex Gr-Ex), Université Sorbonne Paris Cité (USPC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Centre de référence maladie rare des cytopénies auto-immunes de l'adulte (GECAI - Hôpital Henri-Mondor - UPEC), IMRB - CEPIA/'Clinical Epidemiology And Ageing : Geriatrics, Primary Care and Public Health' [Créteil] (U955 Inserm - UPEC), Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Service Hématologie - IUCT-Oncopole [CHU Toulouse], Pôle Biologie [CHU Toulouse], Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Pôle IUCT [CHU Toulouse], Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Institut Universitaire du Cancer de Toulouse - Oncopole (IUCT Oncopole - UMR 1037), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre hospitalier universitaire de Poitiers (CHU Poitiers), Université de Brest (UBO), Centre Hospitalier Régional Universitaire de Brest (CHRU Brest), CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN), Hôpital Haut-Lévêque [CHU Bordeaux], CHU Bordeaux [Bordeaux], Université de Bordeaux (UB), Hôpital Jean Verdier [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Service de Médecine Interne [CHU Rouen], CHU Rouen, Normandie Université (NU)-Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU), Hopital Saint-Louis [AP-HP] (AP-HP), Centre hospitalier universitaire de Nantes (CHU Nantes), Institute for Translational Research in Inflammation - U 1286 (INFINITE (Ex-Liric)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Service de Médecine Interne - Immunologie Clinique [AP-HP Bicêtre], AP-HP Hôpital Bicêtre (Le Kremlin-Bicêtre), Service d'Hématologie biologique [CHU Limoges], and CHU Limoges

Subjects
Purpura, Thrombocytopenic, Idiopathic, Immunology, Pregnancy Complications, Hematologic, Infant, Newborn, Cell Biology, Hematology, Biochemistry, Cohort Studies, Thrombocytopenia, Neonatal Alloimmune, Pregnancy, Humans, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, Female, Prospective Studies, Retrospective Studies
Abstract

The risk of immune thrombocytopenia (ITP) worsening during pregnancy and neonatal ITP (NITP) have never been prospectively studied. We included 180 pregnant and 168 nonpregnant women with ITP in a prospective, multicenter, observational cohort study. A total of 131 pregnant women with ITP were matched to 131 nonpregnant women with ITP by history of splenectomy, ITP status (no response, response, complete response), and duration. Groups were followed for 15 months. The primary outcome was the first occurrence of ITP worsening defined by a composite end point including bleeding events and/or severe thrombocytopenia (

Published
2022
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2. Belatacept in renal transplant recipient with mild immunologic risk factor: A pilot prospective study (BELACOR)

Author

Laurent Salomon, Stéphanie Malard, Thomas Robert, Caroline Pilon, Emmanuelle Boutin, Florence Canoui-Poitrine, Antoine Durrbach, Marie Matignon, Anissa Zarour, Claire Leibler, Anissa Moktefi, Philippe Grimbert, Chloé Samson, Pierre-André Natella, Institut Mondor de Recherche Biomédicale (IMRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), service Anatomie et Cytologie Pathologique, Hôpital Henri Mondor, Assistance Publique - Hôpitaux de Paris (AP-HP), Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Hopital Saint-Louis [AP-HP] (AP-HP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), IMRB - CEPIA/'Clinical Epidemiology And Ageing : Geriatrics, Primary Care and Public Health' [Créteil] (U955 Inserm - UPEC), Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Groupe Henri Mondor-Albert Chenevier, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Hôpital Albert Chenevier, Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), CHU Necker - Enfants Malades [AP-HP], Clinical Research Unit (URC Mondor), Hôpitaux Universitaires Henri-Mondor AP-HP, Néphrologie [CHU Bicêtre], AP-HP Hôpital Bicêtre (Le Kremlin-Bicêtre), Laboratoire de Biomécanique et Mécanique des Chocs (LBMC UMR T9406), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut Français des Sciences et Technologies des Transports, de l'Aménagement et des Réseaux (IFSTTAR), Service de néphrologie, dialyse, aphérèses et transplantation, and CHU Grenoble-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])

Subjects
Graft Rejection, Male, Pilot Projects, kidney transplantation/nephrology, immunosuppression/immune modulation, 030230 surgery, Kidney Function Tests, law.invention, Postoperative Complications, 0302 clinical medicine, Randomized controlled trial, Isoantibodies, Risk Factors, law, Immunology and Allergy, Medicine, Pharmacology (medical), Prospective Studies, immunosuppressant - fusion proteins and monoclonal antibodies, Prospective cohort study, belatacept, Incidence (epidemiology), Graft Survival, clinical trial, Middle Aged, Prognosis, Female, rejection, Immunosuppressive Agents, Glomerular Filtration Rate, medicine.drug, medicine.medical_specialty, Urology, Renal function, clinical research/practice, Belatacept, Abatacept, 03 medical and health sciences, antibody-mediated (ABMR), panel reactive antibody (PRA), Humans, Risk factor, Retrospective Studies, Transplantation, business.industry, Kidney Transplantation, Calcineurin, Clinical trial, Kidney Failure, Chronic, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, business, Follow-Up Studies
Abstract

The benefit of belatacept on antibody-mediated rejection (ABMR) incidence after kidney transplant with preformed donor-specific antibodies (DSAs) has never been assessed. Between 2014 and 2016, we conducted a multicenter prospective clinical trial with 49 patients to determine kidney allograft outcome in recipients with preformed DSAs (maximal mean fluorescence intensity 500 to 3000) treated with belatacept (BELACOR trial). Immunosuppressive strategy included antithymocyte globulin, belatacept, mycophenolate mofetil, and steroids. An ancillary control group was designed retrospectively, including patients fulfilling the same inclusion criteria treated with calcineurin inhibitors. In BELACOR group, no patient exhibited acute ABMR, patient and allograft survival at 1 year was 100% and 95.4%, respectively, and the estimated glomerular filtration rate was 53.2 mL/min/1.73 m2 . However, the 12-month incidence of acute T cell-mediated rejection was 25.4% (14.5% to 42.4%). Comparison with the control group showed significantly higher T cell-mediated rejection incidence only in the BELACOR group (P = .003). Considering the DSAs, the outcome was similar in the 2 groups except a significantly higher number of patients displayed a complete disappearance of class II DSAs in the BELACOR group (P = .001). Belatacept was not associated with an acute ABMR increased risk and may be considered as immunosuppressive strategy in transplant recipients with preformed DSAs (maximal mean fluorescence intensity 500 to 3000). Prospective randomized trials are needed to confirm these results.

Published
2019
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3. Development of Biphasic Formulations for Use in Electrowetting-Based Liquid Lenses with a High Refractive Index Difference

Author

Géraldine Malet, Matthias S. Ober, Mathieu Maillard, Caroline Woelfle-Gupta, Franck Amiot, Benjamin Burger, Bruno Berge, Daniel L. Dermody, Department of Chemistry, University of Freiburg [Freiburg], Laboratoire des Multimatériaux et Interfaces (LMI), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Service de Génomique Fonctionnelle, and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)

Subjects
Physics::Optics, 02 engineering and technology, 010402 general chemistry, Curvature, 01 natural sciences, Phase Transition, law.invention, Physics::Fluid Dynamics, Optics, law, Phase (matter), Optical medium, Transition Temperature, Electrical conductor, ComputingMilieux_MISCELLANEOUS, Lenses, Viscosity, Chemistry, business.industry, Water, Equipment Design, General Chemistry, General Medicine, [CHIM.MATE]Chemical Sciences/Material chemistry, 021001 nanoscience & nanotechnology, 0104 chemical sciences, Lens (optics), Refractometry, Electrowetting, 0210 nano-technology, business, Hydrophobic and Hydrophilic Interactions, Oils, Refractive index, Voltage
Abstract

Commercial electrowetting-based liquid lenses are optical devices containing two immiscible liquids as an optical medium. The first phase is a droplet of a high refractive index oil phase placed in a ring-shaped chassis. The second phase is electrically conductive and has a similar density over a wide temperature range. Droplet curvature and refractive index difference of two liquids determine the optical strength of the lens. Liquid lenses take advantage of the electrowetting effect, which induces a change of the interface's curvature by applying a voltage, thereby providing a variable focal that is useful in autofocus applications. The first generation of lens modules were highly reliable, but the optical strength and application scope was limited by a low refractive index difference between the oil and conductive phase. Described herein is an effort to increase the refractive index difference between both phases, while maintaining other critical application characteristics of the liquids, including a low freezing point, viscosity, phase miscibility, and turbidity after thermal shock. An important challenge was the requirement that both phases have to have matching densities and hence had to be optimized simultaneously. Using high throughput experimentation in conjunction with statistical design of experiments (DOE), we have developed a series of empirical models to predict multiple physicochemical properties of both phases and derived ideal locations within the formulation space. This approach enabled the development of reliable liquid lenses with a previously unavailable refractive index difference of Δ n

Published
2018
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4. A Tandem of SH3-like Domains Participates in RNA Binding in KIN17, a Human Protein Activated in Response to Genotoxics

Author

Joël Couprie, Sophie Zinn-Justin, Enrico A. Stura, Ghislaine Pinon-Lataillade, M. Schiltz, Muriel Gondry, Albane le Maire, Jaime F. Angulo, Mireille Moutiez, Département d'Ingenierie et d'Etudes des Protéines (DIEP), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Ecole Polytechnique Fédérale de Lausanne (EPFL), Institut des Sciences du Vivant Frédéric JOLIOT (JOLIOT), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Service d'Ingénierie Moléculaire pour la Santé (ex SIMOPRO) (SIMoS), Médicaments et Technologies pour la Santé (MTS), Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE)-Université Paris-Saclay-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), and CEA, DSV, DRR, Service de Génomique Fonctionnelle

Subjects
RNA-binding protein, Biology, DNA-binding protein, src Homology Domains, Structural Biology, Humans, SH3-like fold, Binding site, Nuclear protein, Molecular Biology, Gene, Regulation of gene expression, Binding Sites, [SDV.BBM.BS]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Structural Biology [q-bio.BM], KIN17 protein, Lysine, Nuclear Proteins, RNA-Binding Proteins, RNA, Tungsten Compounds, RNA binding, Cell biology, DNA-Binding Proteins, Gene Expression Regulation, Biochemistry, tungstate, TAF4, X-ray structure, Mutagens
Abstract

International audience; The human KIN17 protein is an essential nuclear protein conserved from yeast to human and expressed ubiquitously in mammals. Suppression of Rts2, the yeast equivalent of gene KIN17, renders the cells unviable, and silencing the human KIN17 gene slows cell growth dramatically. Moreover, the human gene KIN17 is up-regulated following exposure to ionizing radiations and UV light, depending on the integrity of the human global genome repair machinery. Its ectopic over-expression blocks S-phase progression by inhibiting DNA synthesis. The C-terminal region of human KIN17 is crucial for this anti-proliferation effect. Its high-resolution structure, presented here, reveals a tandem of SH3-like subdomains. This domain binds to ribonucleotide homopolymers with the same preferences as the whole protein. Analysis of its structure complexed with tungstate shows structural variability within the domain. The interaction with tungstate is mediated by several lysine residues located within a positively charged groove at the interface between the two subdomains. This groovecould be the site of interaction with RNA, since mutagenesis of two of these highly conserved lysine residue weakens RNA binding

Published
2006
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5. Update on infectious risks associated with dental unit waterlines

Author

Amélie Robert, Vanessa Barbot, Christine Imbert, Marie-Hélène Rodier, Microbiologie de l'Eau (MDE), Ecologie et biologie des interactions (EBI), Université de Poitiers-Centre National de la Recherche Scientifique (CNRS)-Université de Poitiers-Centre National de la Recherche Scientifique (CNRS), Service de Génomique Fonctionnelle, and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)

Subjects
Microbiology (medical), Dental Instruments, Immunology, Dentistry, Biology, Microbial contamination, Microbiology, Dental treatments, Risk Assessment, 03 medical and health sciences, 0302 clinical medicine, Immunology and Allergy, Humans, 0303 health sciences, Protozoan Infections, 030306 microbiology, business.industry, Dental Equipment, Dental procedures, 030206 dentistry, General Medicine, Bacterial Infections, 6. Clean water, 3. Good health, stomatognathic diseases, Infectious Diseases, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Mycoses, Virus Diseases, Biofilms, Dental chair, Infectious risk, business, Water Microbiology
Abstract

International audience; Modern dental chair units consist of a network of interconnected narrow-bore plastic tubes called dental unit waterlines (DUWLs). The water delivered by these DUWLs acts as both a coolant for a range of instruments and an irrigant during dental treatments. The quality of water is of considerable importance because both patients and dental team are regularly exposed to water and aerosols generated by dental equipment. Studies have demonstrated that DUWLs provide a favourable environment for microbial proliferation and biofilm formation, and that water is consequently often contaminated with high densities of various microorganisms (bacteria, fungi, protozoa, viruses). The presence of high levels of microbial contamination may be a health problem for dentists and patients, especially those who are immunocompromised. The current status of knowledge on microbial contamination of DUWLs is presented, with an emphasis on the infectious risk associated with DUWLs and on the various approaches for disinfecting and protecting DUWLs.

Published
2012
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6. The hyperscape project: Participative game informational construction

Author

Woloszyn, Philippe, Bourdin, G., Girardot, Jean-Jacques, Service de Génomique Fonctionnelle, and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)

Subjects
[SHS.HISPHILSO]Humanities and Social Sciences/History, Philosophy and Sociology of Sciences, [SHS.HISPHILSO] Humanities and Social Sciences/History, Philosophy and Sociology of Sciences, territorial Intelligence, ComputingMilieux_MISCELLANEOUS, Intelligence territoriale
Abstract

International audience

Published
2009

7. The Hyperscape project: the Landscapes Observatory. Tool Coproduction and Acknowledgment Participative Challenge-Games

Author

Woloszyn, Philippe, Bourdin, G., Girardot, Jean-Jacques, Service de Génomique Fonctionnelle, and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)

Subjects
[SHS.HISPHILSO]Humanities and Social Sciences/History, Philosophy and Sociology of Sciences, Territorial Intelligence, [SHS.HISPHILSO] Humanities and Social Sciences/History, Philosophy and Sociology of Sciences, ComputingMilieux_MISCELLANEOUS, Intelligence territoriale
Abstract

International audience

Published
2009

8. Broad modulation of gene expression in CD4+ lymphocyte subpopulations in response to low doses of ionizing radiation

Author

Aurélie Vaurijoux, Peggy Maltère, Gaëtan Gruel, Cyrille Petat, Xavier Gidrol, Philippe Voisin, Pascale Voisin, Sandrine Roch-Lefèvre, Eric Gregoire, Laurence Roy, Laboratoire de dosimétrie biologique (DRPH/SRBE/LDB), Institut de Radioprotection et de Sûreté Nucléaire (IRSN), Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Laboratoire de dosimétrie biologique (IRSN/DRPH/SRBE/LDB), Service de RadioBiologie et d'Epidémiologie (IRSN/DRPH/SRBE), and Institut de Radioprotection et de Sûreté Nucléaire (IRSN)-Institut de Radioprotection et de Sûreté Nucléaire (IRSN)

Subjects
CD4-Positive T-Lymphocytes, Male, cycline, protein synthesis, Lymphocyte, [SDV]Life Sciences [q-bio], complementary DNA, Ionizing radiation, cyclin dependent kinase inhibitor 1A, low energy radiation, human experiment, 0302 clinical medicine, Gene expression, Cells, Cultured, Whole blood, Regulation of gene expression, 0303 health sciences, Cultured, Radiation, adult, article, CD8 antigen, Middle Aged, protein DDB2, unclassified drug, medicine.anatomical_structure, priority journal, 030220 oncology & carcinogenesis, CD4 antigen, ionizing radiation, protein Bax, radiation response, Cells, CD56 antigen, Biophysics, oxidative phosphorylation, Biology, Radiation Dosage, Dose-Response Relationship, 03 medical and health sciences, medicine, Humans, Radiology, Nuclear Medicine and imaging, controlled study, Irradiation, human, normal human, protein expression, 030304 developmental biology, growth arrest and DNA damage inducible protein 45, human cell, DNA microarray, CD4 lymphocyte count, cell type, Dose-Response Relationship, Radiation, Molecular biology, human tissue, Gene Expression Regulation, Gene chip analysis, RNA, lymphocyte subpopulation, CD8
Abstract

To compare me responses of the different lymphocyte sub-types after an exposure of whole blood to low doses of ionizing radiation, we examined variations in gene expression in different lymphocyte subpopulations using microarray technology. Blood samples from five healthy donors were independently exposed to 0 (sham irradiation), 0.05 and 0.5 Gy of ionizing radiation. Three and 24 h after exposure, CD56+, CD4+ and CD8+ cells were negatively isolated. RNA from each set of experimental conditions was competitively hybridized on 25k oligonucleotide microarrays. Modifications of gene expression were measured after both intervals and in all cell types. Twenty-four hours after exposure to 0.5 Gy, we observed an induction of the expression of BAX, PCNA, GADD45, DDB2 and CDKNIA. However, the numbers of modulated genes greatly differed between cell types. In particular, 3 h after exposure to doses as low as 0.05 Gy, the number of down-modulated genes was 10 times greater for CD4+ cells than for all other cell types. Moreover, most of these repressed genes were taking part in the cell processes of protein biosynthesis and oxidative phosphorylation. The results suggest that several biological pathways in CD4+ cells could be sensitive to low doses of radiation. Therefore, specifically studying CD4+ cells could help to understand the mechanisms involved in low-dose response and allow their detection.

Published
2007
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9. Sensitization to the conditioned rewarding effects of morphine modulates gene expression in rat hippocampus

Author

Xavier Gidrol, Amelie Robert, Florence Noble, Cindie Courtin, Bernard P. Roques, Cynthia Marie-Claire, Neuropsychopharmacologie des addictions. Vulnérabilité et variabilité expérimentale et clinique (NAVVEC - UM 81 (UMR 8206/ U705)), Université Paris Diderot - Paris 7 (UPD7)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut de Recherche pour le Développement (IRD)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Pharmacochimie moléculaire et structurale, Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), MILDT AH020G, Marie-Claire, Cynthia, Neuropsychopharmacologie des addictions. Vulnérabilité et variabilité expérimentale et clinique ( NAVVEC (UM 81) ), Université Paris Diderot - Paris 7 ( UPD7 ) -Institut des sciences du Médicament -Toxicologie - Chimie - Environnement ( IFR71 ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL ( ENSCP ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Ecole Nationale Supérieure de Chimie de Paris- Chimie ParisTech-PSL ( ENSCP ) -Centre National de la Recherche Scientifique ( CNRS ) -Institut de Recherche pour le Développement ( IRD ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), Institut des sciences du Médicament -Toxicologie - Chimie - Environnement ( IFR71 ), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5)-Institut des sciences du Médicament -Toxicologie - Chimie - Environnement (IFR71), Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Université Paris Diderot - Paris 7 (UPD7), and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Paris Descartes - Paris 5 (UPD5)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Male, MESH: Hippocampus, MESH : Narcotics, Hippocampus, MESH: Conditioning, Operant, MESH : Analysis of Variance, MESH: Rats, Sprague-Dawley, MESH : Hippocampus, Pharmacology, Hippocampal formation, Rats, Sprague-Dawley, chemistry.chemical_compound, 0302 clinical medicine, Gene expression, MESH: Animals, Neurotransmitter, Sensitization, Regulation of gene expression, 0303 health sciences, Morphine, MESH : Gene Expression Regulation, MESH : Rats, MESH: Gene Expression Regulation, MESH: Narcotics, MESH : Microarray Analysis, medicine.anatomical_structure, [ SDV.BBM.GTP ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Narcotics, MESH: Rats, MESH : Male, MESH : Drug Administration Schedule, Biology, MESH: Drug Administration Schedule, Drug Administration Schedule, Article, MESH : Conditioning, Operant, 03 medical and health sciences, Cellular and Molecular Neuroscience, Reward, MESH: Analysis of Variance, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], medicine, Animals, 030304 developmental biology, MESH: Reward, Analysis of Variance, Microarray analysis techniques, MESH : Morphine, Microarray Analysis, MESH : Rats, Sprague-Dawley, MESH: Male, Rats, MESH: Microarray Analysis, MESH: Morphine, Gene Expression Regulation, chemistry, Suppression subtractive hybridization, Conditioning, Operant, MESH : Animals, MESH : Reward, 030217 neurology & neurosurgery
Abstract

Opiates addiction is characterized by its long-term persistence. In order to study the enduring changes in long-term memory in hippocampus, a pivotal region for this process, we used suppression subtractive hybridization to compare hippocampal gene expression in morphine and saline-treated rats. Animals were subjected to an extended place preference paradigm consisting of four conditioning phases. Sensitization to the reinforcing effects of the drug occurred after three conditioning phases. After 25 days of treatment rats were euthanized and the complementary DNA (cDNA) from the hippocampus of morphine-dependent and saline-treated animals were then screened for differentially expressed cDNAs. The selected 177 clones were then subjected to a microarray procedure and 20 clones were found differentially regulated. The pattern of regulated genes suggests impairments in neurotransmitter release and the activation of neuroprotective pathways.

Published
2007
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10. Novel Microarray-Based Method for Estimating Exposure to Ionizing Radiation

Author

Gaëtan Gruel, Alexandre Pawlik, Olivier Alibert, Beatrice Jacquelin, Xavier Gidrol, Carlo Lucchesi, Anissa Zarour, Diana Tronik-LeRoux, Thierry Kortulewski, Vincent Frouin, Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), This work was supported by funds from the Commisariat à l'Energie Atomique (CEA) and Electricité de France (EDF), GG was also supported by a predoctoral grant of the Association pour la Recherche sur le Cancer (ARC), AP by a predoctoral grant of the Fondation pour la Recherche Médicale (FRM), and BJ by a postdoctoral training grant from Genopole©-Evry., and We would like to thank C. Joubert, P Flamant and V. Neuville for outstanding work in animal care, G. Munhoz for helpful discussions, and P. Soularue, A. Robert and P. Maltere for producing the cDNA microarray slides.

Subjects
Male, Microarray, Gene Expression, Ionizing radiation, Mice, 0302 clinical medicine, Radiation, Ionizing, MESH: Radiometry / methods, MESH: Body Burden, Gene expression, MESH: Animals, Oligonucleotide Array Sequence Analysis, 0303 health sciences, Radiation, MESH: Relative Biological Effectiveness, Blood Proteins, 030220 oncology & carcinogenesis, Body Burden, Algorithms, MESH: Radiation Dosage, Biophysics, Computational biology, Biology, MESH: Radiation, Ionizing, Radiation Dosage, 03 medical and health sciences, MESH: Mice, Inbred C57BL, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], MESH: Dose-Response Relationship, Radiation, Animals, MESH: Gene Expression / radiation effects, Radiology, Nuclear Medicine and imaging, Epigenetics, Radiometry, Gene, MESH: Mice, 030304 developmental biology, Dosimeter, business.industry, Dose-Response Relationship, Radiation, Environmental Exposure, MESH: Male, Mice, Inbred C57BL, Gene selection, MESH: Environmental Exposure / analysis, MESH: Algorithms, Gene chip analysis, MESH: Blood Proteins / analysis, MESH: Oligonucleotide Array Sequence Analysis / methods, Nuclear medicine, business, Relative Biological Effectiveness
Abstract

International audience; Accurate estimation of the dose of ionizing radiation to which individuals have been exposed is critical for therapeutic treatment. We investigated whether gene expression profiles could be used to evaluate the dose received, thereby serving as a biological dosimeter. We used cDNA microarrays to monitor changes in gene expression profiles induced by ionizing radiation in mouse total blood. The subsets of genes best characterizing each dose were identified by resampling the original data set and calculating the intersection of the dose signatures. This analytical strategy minimizes the impact of potential genetic/epigenetic variation between mice and overcomes the bias in gene selection inherent to microarray technology. The significance of the identified signatures was evaluated by monitoring the type I error rate by in silico negative control simulation. Based on the distribution of the mean ratios of the selected probes, we were able to identify transcription profiles giving 83% to 100% correct estimation of the dose received by test mice, demonstrating that the selected probes could be used to determine the dose of radiation to which the animals had been exposed. This method could potentially be generalized to determine the level of exposure to other toxins and could be used to develop new related clinical applications.

Published
2006
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11. Vacuum Casting to Manufacture a Plastic Biochip for Highly Parallel Cell Transfection

Author

M Denoual, X Gidrol, Pascal Mognol, B. Le Pioufle, Y Macé, D Castel, Systèmes et Applications des Technologies de l'Information et de l'Energie (SATIE), École normale supérieure - Cachan (ENS Cachan)-Université Paris-Sud - Paris 11 (UP11)-Institut Français des Sciences et Technologies des Transports, de l'Aménagement et des Réseaux (IFSTTAR)-École normale supérieure - Rennes (ENS Rennes)-Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine-Conservatoire National des Arts et Métiers [CNAM] (CNAM)-Centre National de la Recherche Scientifique (CNRS), Institut de Recherche en Communications et en Cybernétique de Nantes (IRCCyN), Mines Nantes (Mines Nantes)-École Centrale de Nantes (ECN)-Ecole Polytechnique de l'Université de Nantes (EPUN), Université de Nantes (UN)-Université de Nantes (UN)-PRES Université Nantes Angers Le Mans (UNAM)-Centre National de la Recherche Scientifique (CNRS), Bio-MIcroSystèmes et BioSensors (SATIE-BIOMIS), Systèmes d'Information et d'Analyse Multi-Echelles (SIAME), Université Paris-Seine-Université Paris-Seine-Conservatoire National des Arts et Métiers [CNAM] (CNAM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Cachan (ENS Cachan)-Université Paris-Sud - Paris 11 (UP11)-Institut Français des Sciences et Technologies des Transports, de l'Aménagement et des Réseaux (IFSTTAR)-École normale supérieure - Rennes (ENS Rennes)-Université de Cergy Pontoise (UCP), Université Paris-Seine-Université Paris-Seine-Conservatoire National des Arts et Métiers [CNAM] (CNAM)-Centre National de la Recherche Scientifique (CNRS)-Systèmes et Applications des Technologies de l'Information et de l'Energie (SATIE), CEA, DSV, DRR, Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Université Paris-Seine-Université Paris-Seine-Conservatoire National des Arts et Métiers [CNAM] (CNAM), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-Centre National de la Recherche Scientifique (CNRS), Systèmes d'Information et d'Analyse Multi-Echelles (SATIE-SIAME), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Cachan (ENS Cachan)-Université Paris-Sud - Paris 11 (UP11)-Institut Français des Sciences et Technologies des Transports, de l'Aménagement et des Réseaux (IFSTTAR)-École normale supérieure - Rennes (ENS Rennes)-Université de Cergy Pontoise (UCP), and HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-Centre National de la Recherche Scientifique (CNRS)-Systèmes et Applications des Technologies de l'Information et de l'Energie (SATIE)

Subjects
Rapid prototyping, Materials science, Applied Mathematics, Vacuum casting, 010401 analytical chemistry, Thermosetting polymer, Nanotechnology, 02 engineering and technology, 021001 nanoscience & nanotechnology, Chip, 01 natural sciences, 0104 chemical sciences, Casting (metalworking), Microsystem, [SPI.NANO]Engineering Sciences [physics]/Micro and nanotechnologies/Microelectronics, 0210 nano-technology, Biochip, Instrumentation, Engineering (miscellaneous), Microfabrication
Abstract

International audience; A novel polymer microarray fabrication technique is presented and applied to the realization of a biochip for highly parallelized cell transfection. The proposed microfabrication technique is derived from a macroscale rapid prototyping technique called vacuum casting. It was optimized to reduce production cost, in order to produce small series (100-10 000 chip series) of chips to meet demand in today's market of cellulomics. Microfabrication technologies and rapid prototyping technologies are combined to shape the master part, which can thus involve microsized features. The corresponding female structure is moulded in a flexible silicone material. The duplicated polymer chips are obtained by casting a thermosetting plastic under vacuum. The dimensional replication accuracy between the master part and the duplicated parts is uniform over the duplicated parts and better than 1%. Advantages of the proposed technique over existing plastic microfabrication techniques are discussed in the paper. Using this microfabrication technique, we produced a plastic biochip for highly parallelized transfection of arrays of living cells. The feasibility of parallel lipofection was demonstrated: two different plasmids encoding, respectively, eGFP and DsRED2 were inserted into HEK293T cells. The transfection was monitored through fluorescence observation after 72 h showing successful expression of both genes.

Published
2006
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12. An open-access long oligonucleotide microarray resource for analysis of the human and mouse transcriptomes

Author

Philippe Kastner, Philippe Dessen, Christine Bole-Feysot, Paul A. Lyons, Virginie Virolle, Philippe Rostagno, Franck Amiot, Gary Williams, Chimène Moreilhon, Pascal Barbry, Bernard Jost, Roslin Russell, Erdogan Gulari, Xavier Gidrol, Géraldine Rios, Virginie Defamie, Kevin Le Brigand, Virginie Magnone, Tom C. Freeman, Jean Marie Rouillard, Bernard Mari, Institut de pharmacologie moléculaire et cellulaire (IPMC), Université Nice Sophia Antipolis (1965 - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS), MRC Rosalind Franklin Centre for Genomics Research, Department of Chemical Engineering, University of Michigan [Ann Arbor], University of Michigan System-University of Michigan System, Biodiscovery LLC, Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Laboratoire de Génétique Oncologique, Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Réseau National des Génopoles, the GIP Aventis, the French Ministry of Industry (Réseau GenHomme), the French Association Vaincre la Mucoviscidose, the INCA, the CNRS, the UK Medical Research Council, Université Nice Sophia Antipolis (... - 2019) (UNS), and Kervella, Jacqueline

Subjects
Microarray, Transcription, Genetic, MESH: Oligonucleotide Probes, In silico, [SDV.GEN] Life Sciences [q-bio]/Genetics, Biology, MESH: Databases, Nucleic Acid, MESH: Expressed Sequence Tags, Transcriptome, 03 medical and health sciences, MESH: Gene Expression Profiling, Mice, 0302 clinical medicine, Genetics, Animals, Humans, MESH: Animals, MESH: Mice, 030304 developmental biology, Oligonucleotide Array Sequence Analysis, Expressed Sequence Tags, 0303 health sciences, Expressed sequence tag, [SDV.GEN]Life Sciences [q-bio]/Genetics, Internet, MESH: Humans, Oligonucleotide, MESH: Transcription, Genetic, Gene Expression Profiling, Gene expression profiling, MESH: Internet, Oligonucleotide Microarray, 030220 oncology & carcinogenesis, MESH: Oligonucleotide Array Sequence Analysis, Methods Online, DNA microarray, Databases, Nucleic Acid, Oligonucleotide Probes
Abstract

Two collections of oligonucleotides have been designed for preparing pangenomic human and mouse microarrays. A total of 148,993 and 121,703 oligonucleotides were designed against human and mouse transcripts. Quality scores were created in order to select 25,342 human and 24,109 mouse oligonucleotides. They correspond to: (i) a BLAST-specificity score; (ii) the number of expressed sequence tags matching each probe; (iii) the distance to the 3' end of the target mRNA. Scores were also used to compare in silico the two microarrays with commercial microarrays. The sets described here, called RNG/MRC collections, appear at least as specific and sensitive as those from the commercial platforms. The RNG/MRC collections have now been used by an Anglo-French consortium to distribute more than 3500 microarrays to the academic community. Ad hoc identification of tissue-specific transcripts and a approximately 80% correlation with hybridizations performed on Affymetrix GeneChiptrade mark suggest that the RNG/MRC microarrays perform well. This work provides a comprehensive open resource for investigators working on human and mouse transcriptomes, as well as a generic method to generate new microarray collections in other organisms. All information related to these probes, as well as additional information about commercial microarrays have been stored in a freely-accessible database called MEDIANTE.

Published
2006
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13. Therapeutic effect of human mesenchymal stem cells in skin after radiation damage [Potentiel thérapeutique des cellules souches mésenchymateuses humaines dans les lésions cutanées radioinduites]

Author

Bensidhoum, M., Gobin, S., alain chapel, Lemaitre, G., Bouet, S., Waksman, G., Thierry, D., Martin, M. T., CEA, DSV, DRR, Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Laboratoire de Génomique et Radiobiologie du Kératinocyte (LGRK), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE), Laboratoire de Thérapie Cellulaire et de Radioprotection Accidentelle (IRSN/DRPH/SRBE/LTCRA), Service de RadioBiologie et d'Epidémiologie (IRSN/DRPH/SRBE), Institut de Radioprotection et de Sûreté Nucléaire (IRSN)-Institut de Radioprotection et de Sûreté Nucléaire (IRSN), Laboratoire de radiobiologie et d'étude du génome (LREG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Laboratoire de Thérapie Cellulaire et de Radioprotection Accidentelle (DRPH/SRBE/LTCRA), and Institut de Radioprotection et de Sûreté Nucléaire (IRSN)

Subjects
[SDV]Life Sciences [q-bio]
Abstract

Over 50 % of all cancer patients presently receive radiotherapy at one stage in their treatment course. Inevitably skin is one of the most frequently damaged tissue due to its localization and constant turn-over. Our present goal is to reduce radiation-induced complications in human skin through stem cell therapy, particulary in human epidermis. Mesenchymal Stem Cells (MSCs) have been shown to be multipotent cells able to engraft in many tissues after injury. Herein, we isolated human MSCs and tested their capability to improve skin wound healing after irradiation. This potential was assessed in NOD/SCID mice which received 30 Gy locally on the thigh. This dose caused within 3 weeks local epidermis necrosis which was repaired within 13 weeks. MSCs were intravenously injected in irradiated mice 24 hours after exposure. Clinical scoring throughout 6 weeks gave indications that human MSCs reduced the extent of damage and accelerated the wound healing process. We show by quantitative qPCR and histological studies the presence of human MSCs derived cells into the scar. Human MSCs homed to the damaged skin and participated to the wound healing process. These results open prospects for cellular therapy by MSCs in irradiated epithelial tissues and could be extended to the whole general field of cutaneous cicatrization, particularly after burns.

Published
2005
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14. Novel pathway for megakaryocyte production after in vivo conditional eradication of integrin αIIb-expressing cells

Author

Thierry Kortulewski, Pierre Vaigot, Pascal Soularue, Olivier Alibert, Alexandre Pawlik, Diana Tronik-Le Roux, Beatrice Jacquelin, Xavier Gidrol, Christophe Joubert, Gaëtan Gruel, Laboratoire de Cancérologie Expérimentale (LCE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay, Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Supported by funds from the Commissariat à l'Energie Atomique (CEA) and the Association Française contre les Myopathies (AFM) and by a postdoctoral training grant from Genopole-Evry., and We thank Drs A. Galy and E. Lauret for their assistance in cell culture studies, J.-M. Egly and Y. Senis for critical reading of the manuscript, and Dr G. Marguerie, who initially established the CEA microarray platform. We also thank the staff of the Genethon mouse facility and P. Flament and V. Neuville for outstanding work in animal care at the CEA-FAR mouse facility. All the animal studies reported were performed in accordance with current French regulations (Décret no. 87-848 modifié, Ministère de l'Agriculture).

Subjects
Myeloid, Cellular differentiation, Biochemistry, Mice, 0302 clinical medicine, Megakaryocyte, Myeloid Cells, MESH: Animals, MESH: Proteins, Lymphocytes, MESH: Platelet Membrane Glycoprotein IIb* / genetics, 0303 health sciences, education.field_of_study, MESH: Bone Marrow Cells / physiology, Polycomb Repressive Complex 2, MESH: Regeneration, Hematology, 3. Good health, Cell biology, DNA-Binding Proteins, Haematopoiesis, medicine.anatomical_structure, Myelopoiesis, Megakaryocytes, MESH: Polycomb Repressive Complex 2, Platelet Membrane Glycoprotein IIb, MESH: DNA Nucleotidylexotransferase / genetics, MESH: Mice, Transgenic, Immunology, Population, Bone Marrow Cells, Mice, Transgenic, MESH: Lymphocytes / physiology, Biology, MESH: Hematopoiesis, 03 medical and health sciences, MESH: Gene Expression Profiling, DNA Nucleotidylexotransferase, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], MESH: Megakaryocytes / cytology, medicine, MESH: Enhancer of Zeste Homolog 2 Protein, [SDV.MHEP.PHY]Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], Animals, Regeneration, Cell Lineage, Enhancer of Zeste Homolog 2 Protein, Lymphopoiesis, education, MESH: Mice, MESH: Myeloid Cells / physiology, 030304 developmental biology, MESH: DNA-Binding Proteins / genetics, Gene Expression Profiling, MESH: Histone-Lysine N-Methyltransferase, Proteins, Histone-Lysine N-Methyltransferase, Cell Biology, MESH: Cell Lineage, Molecular biology, Hematopoiesis, Gene expression profiling, 030215 immunology
Abstract

Our knowledge of the molecular mechanisms that regulate hematopoiesis in physiologic and pathologic conditions is limited. Using a molecular approach based on cDNA microarrays, we demonstrated the emergence of an alternative pathway for mature bone marrow cell recovery after the programmed and reversible eradication of CD41+ cells in transgenic mice expressing a conditional toxigene targeted by the platelet αIIb promoter. The expression profile of the newly produced CD41+ cells showed high levels of transcripts encoding Ezh2, TdT, Rag2, and various immunoglobulin (Ig) heavy chains. In this context, we identified and characterized a novel population of Lin-Sca-1hic-Kit- cells, with a lymphoid-like expression pattern, potentially involved in the reconstitution process. Our study revealed novel transcriptional cross talk between myeloid and lymphoid lineages and identified gene expression modifications that occur in vivo under these particular stress conditions, opening important prospects for therapeutic applications.

Published
2005
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15. Isolement et caractérisation de marqueurs membranaires des cellules progénitrices de l'épiderme humain

Author

Lemaitre, Gilles, Serhal, H., Cohen, Albert, Bourdin, G., Gonnet, Florence, Vaigot, Pierre, Rachidi, Walid, Tortajada, Jeanine, Martin, Michèle, Waksman, Gilles, Gonnet, Florence, Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Analyse et environnement (LAE), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS)

Subjects
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
Abstract

Affiche

Published
2005

16. CD98, a novel marker of transient amplifying human keratinocytes

Author

Xavier Gidrol, Jeanine Tortajada, Gilles Lemaitre, Gilles Waksman, Michèle T. Martin, Florence Gonnet, Pierre Vaigot, Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Analyse et environnement (LAE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS), and Gonnet, Florence

Subjects
Keratinocytes, Caveolin 1, Fusion Regulatory Protein-1, Biology, Biochemistry, Flow cytometry, Cell Line, 03 medical and health sciences, 0302 clinical medicine, Caveolae, medicine, Humans, Immunoprecipitation, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], Molecular Biology, [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], 030304 developmental biology, 0303 health sciences, medicine.diagnostic_test, Epidermis (botany), Flow Cytometry, Immunohistochemistry, Transmembrane protein, 3. Good health, Cell biology, [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM], HaCaT, medicine.anatomical_structure, Membrane protein, 030220 oncology & carcinogenesis, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Electrophoresis, Polyacrylamide Gel, Stem cell, Keratinocyte
Abstract

Identification of plasma membrane markers of basal keratinocytes is essential for sorting basal cells and, subsequently, adult epidermal stem cells. In this study, we isolated caveolin-1-enriched microdomains from human HaCaT keratinocytes and identified proteins representing potential cell surface markers of the epidermis by a proteomic approach. The purification of this caveolae domain allowed us to characterize 53 proteins of which 26% were transmembrane and 32% associated-membrane proteins. One of them, CD98, was found to be co-localized with beta1 integrin at the plasma membrane of the basal keratinocytes of healthy human epidermis. We then isolated CD98-positive keratinocytes from fresh skin biopsies. Using clonogenic assays, we demonstrate that CD98 may be considered as a marker of transient amplifying human keratinocytes.

Published
2005

17. The search of new markers of keratinocyte differentiation

Author

Lemaitre, Gilles, Gonnet, Florence, Lamartine, J., Le Minter, P., Vaigot, Pierre, Gidrol, Xavier, Tortajada, Jeanine, Martin, Michèle, Waksman, Gilles, Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Analyse et environnement (LAE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS), and Gonnet, Florence

Subjects
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
Published
2005

18. Analyse directe par UV-MALDI-TOF de la matrice extra-cellulaire de cellules cultivées sur membrane polyester

Author

Zirah, Séverine, Gonnet, Florence, Lemaitre, Gilles, Waksman, Gilles, Tortajada, Jeanine, Analyse et environnement (LAE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS), Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and Gonnet, Florence

Subjects
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
Abstract

Affiche

Published
2004

19. Prion protein prevents human breast carcinoma cell line from tumor necrosis factor alpha-induced cell death

Author

Geneviève Piétu, Marc Eloit, Hugues Ripoche, Salem Chouaib, Catherine Gaudin, Dominique Dormont, Xavier Pinson, Abdelali Jalil, Samuel Arrabal, Maryam Diarra-Mehrpour, Amandine Pitaval, Institut National de la Santé et de la Recherche Médicale (INSERM), Virologie, École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Agence Française de Sécurité Sanitaire des Aliments (AFSSA), Commissariat à l'énergie atomique et aux énergies alternatives (CEA), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL), Université Paris-Sud - Paris 11 (UP11), Service de Génomique Fonctionnelle, Centre National de la Recherche Scientifique (CNRS), Institut Gustave Roussy (IGR), Génétique oncologique (GO - UMR 8125), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Centre National de la Recherche Scientifique (CNRS), Immunologie intégrative des tumeurs (UMR 1186), Université Paris-Sud - Paris 11 (UP11)-Institut Gustave Roussy (IGR)-Institut National de la Santé et de la Recherche Médicale (INSERM), École nationale vétérinaire - Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA)-Agence Française de Sécurité Sanitaire des Aliments (AFSSA), and École Pratique des Hautes Études (EPHE)

Subjects
EXPRESSION, Cancer Research, Pathology, medicine.medical_specialty, Programmed cell death, DNA, Complementary, [SDV]Life Sciences [q-bio], Cell, Breast Neoplasms, Biology, Transfection, ACTIVATION, PATHWAY, 03 medical and health sciences, 0302 clinical medicine, SIGNALS, Complementary DNA, Cell Line, Tumor, medicine, Cytotoxic T cell, Humans, PrPC Proteins, NEURONS, 030304 developmental biology, Oligonucleotide Array Sequence Analysis, 0303 health sciences, Cell Death, Tumor Necrosis Factor-alpha, Molecular biology, Enzymes, medicine.anatomical_structure, Oncology, Cell culture, Apoptosis, Drug Resistance, Neoplasm, 030220 oncology & carcinogenesis, SURVIVAL, Tumor necrosis factor alpha, Female, TNF-INDUCED CYTOTOXICITY, OVEREXPRESSION, RESISTANCE
Abstract

To define genetic determinants of tumor cell resistance to the cytotoxic action of tumor necrosis factor α (TNF), we have applied cDNA microarrays to a human breast carcinoma TNF-sensitive MCF7 cell line and its established TNF-resistant clone. Of a total of 5760 samples of cDNA examined, 3.6% were found to be differentially expressed in TNF-resistant 1001 cells as compared with TNF-sensitive MCF7 cells. On the basis of available literature data, the striking finding is the association of some differentially expressed genes involved in the phosphatidylinositol-3-kinase/Akt signaling pathway. More notably, we found that the PRNP gene coding for the cellular prion protein (PrPc), was 17-fold overexpressed in the 1001 cell line as compared with the MCF7 cell line. This differential expression was confirmed at the cell surface by immunostaining that indicated that PrPc is overexpressed at both mRNA and protein levels in the TNF-resistant derivative. Using recombinant adenoviruses expressing the human PrPc, our data demonstrate that PrPc overexpression converted TNF-sensitive MCF7 cells into TNF-resistant cells, at least in part, by a mechanism involving alteration of cytochrome c release from mitochondria and nuclear condensation.

Published
2004
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20. Analyse protéomique de fractions membranaires de kératinocytes eucaryotes

Author

Gonnet, Florence, Lemaitre, Gilles, Waksman, Gilles, Tortajada, Jeanine, Analyse et environnement (LAE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS), Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and Gonnet, Florence

Subjects
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
Published
2003

21. Proteomic analysis of caveolin1 enriched fractions of membrane proteins of human keratinocytes

Author

Gonnet, Florence, Lemaitre, Gilles, Gidrol, Xavier, Waksman, Gilles, Tortajada, Jeanine, Gonnet, Florence, Analyse et environnement (LAE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS), Service de Génomique Fonctionnelle, and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)

Subjects
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
Abstract

Affiche

Published
2003

22. Enhanced sequence coverage of proteins interacting with plasma membrane of keratinocytes by using MALDI with different matrices in concert

Author

Gonnet, Florence, Lemaitre, Gilles, Gidrol, Xavier, Waksman, Gilles, Tortajada, Jeanine, Analyse et environnement (LAE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS), Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and Gonnet, Florence

Subjects
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
Abstract

Affiche

Published
2003

23. Identification de protéines membranaires par spectrométrie de masse

Author

Gonnet, Florence, Lemaitre, Gilles, Waksman, Gilles, Tortajada, Jeanine, Analyse et environnement (LAE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS), Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and Gonnet, Florence

Subjects
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
Published
2003

24. Purification of caveolae microdomains from HaCaT keratinocytes followed by proteomic analysis allows the characterization of new markers of human epidermis layers

Author

Lemaitre, Gilles, Gonnet, Florence, Vaigot, Pierre, Gidrol, Xavier, Tortajada, Jeanine, Waksman, Gilles, Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Analyse et environnement (LAE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS), ASBMB, and Gonnet, Florence

Subjects
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
Abstract

Affiche; International audience

Published
2003

25. Etude protéomique d'une fraction membranaire de kératinocytes enrichie en microdomaine caveolae

Author

Gonnet, Florence, Lemaitre, Gilles, Waksman, Gilles, Tortajada, Jeanine, Analyse et environnement (LAE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS), Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and Gonnet, Florence

Subjects
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
Published
2003

26. Identification of genes involved in ceramide-dependent neuronal apoptosis using cDNA arrays

Author

Decraene, C., Brugg, B., Ruberg, M., Eveno, E., Matingou, C., Tahi, F., Jean Mariani, Auffray, C., Pietu, G., Genexpress, Centre National de la Recherche Scientifique (CNRS), Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Neurobiologie des processus adaptatifs (NPA), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), Neurologie Expérimentale et Thérapeutique, IFR70-Institut National de la Santé et de la Recherche Médicale (INSERM), Université d'Évry-Val-d'Essonne (UEVE), This work was supported by the CNRS and grants from European Union to J.M. (TMR projet Neuril) and BIOMED2 programs (EURO-IMAGE Consortium, BMH4-CT-97-2284) to C.A. C.M. was supported by Genome Express and C.D. acknowledges fellowships from the Fédération Française des Groupements Parkinsoniens (FFGP) and the Association pour la Recherche sur le Cancer (ARC)., and Maylin, Françoise

Subjects
MESH: Signal Transduction, MESH: Cell Differentiation, MESH: Rats, MESH: Neurons, Apoptosis, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, PC12 Cells, MESH: Nucleic Acid Hybridization, MESH: Gene Expression Profiling, Sphingosine, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Animals, MESH: PC12 Cells, MESH: Animals, [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], [SDV.BC] Life Sciences [q-bio]/Cellular Biology, Oligonucleotide Array Sequence Analysis, Neurons, Research, Gene Expression Profiling, MESH: Apoptosis, Nucleic Acid Hybridization, Cell Differentiation, MESH: Genes, Rats, Genes, MESH: Oligonucleotide Array Sequence Analysis, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH: Sphingosine, Signal Transduction
Abstract

A cell-culture model was used to establish a profile of gene expression during the effector phase of ceramide-mediated cell death. Of the 239 genes that met the criteria for differential hybridization, 10 correspond to genes previously involved in C2-ceramide or TNF-α signaling pathways and 20 in neuronal disorders, oncogenesis or more broadly in the regulation of proliferation., Background Ceramide is important in many cell responses, such as proliferation, differentiation, growth arrest and apoptosis. Elevated ceramide levels have been shown to induce apoptosis in primary neuronal cultures and neuronally differentiated PC 12 cells. Results To investigate gene expression during ceramide-dependent apoptosis, we carried out a global study of gene expression in neuronally differentiated PC 12 cells treated with C2-ceramide using an array of 9,120 cDNA clones. Although the criteria adopted for differential hybridization were stringent, modulation of expression of 239 genes was identified during the effector phase of C2-ceramide-induced cell death. We have made an attempt at classifying these genes on the basis of their putative functions, first with respect to known effects of ceramide or ceramide-mediated transduction systems, and then with respect to regulation of cell growth and apoptosis. Conclusions Our cell-culture model has enabled us to establish a profile of gene expression during the effector phase of ceramide-mediated cell death. Of the 239 genes that met the criteria for differential hybridization, 10 correspond to genes previously involved in C2-ceramide or TNF-α signaling pathways and 20 in neuronal disorders, oncogenesis or more broadly in the regulation of proliferation. The remaining 209 genes, with or without known functions, constitute a pool of genes potentially implicated in the regulation of neuronal cell death.

Published
2002

27. Isolement et caractérisation de protéines membranaires des kératinocytes

Author

Gonnet, Florence, Lemaitre, Gilles, Waksman, Gilles, Tortajada, Jeanine, Analyse et environnement (LAE), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université d'Évry-Val-d'Essonne (UEVE)-Centre National de la Recherche Scientifique (CNRS), Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and Gonnet, Florence

Subjects
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biochemistry [q-bio.BM], [SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biomolecules [q-bio.BM]
Abstract

Affiche

Published
2002

28. Accumulation of mature mRNA in the nuclear fraction of mammalian cells

Author

Pierre Gondran, François Dautry, Dominique Weil, Franck Amiot, Service de Génomique Fonctionnelle, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Compartimentation et trafic intracellulaire des mRNP = Compartmentation and intracellular traffic of mRNPs (LBD-E14), Laboratoire de Biologie du Développement [IBPS] (LBD), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Paris Seine (IBPS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Paris Seine (IBPS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biologie et de Pharmacologie Appliquée (LBPA), École normale supérieure - Cachan (ENS Cachan)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biologie du Développement [Paris] (LBD), Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Institut de Biologie Paris Seine (IBPS), and Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Cytoplasm, Mature messenger RNA, Biophysics, Biology, Biochemistry, Cell Line, 03 medical and health sciences, mRNA transport, Mice, 0302 clinical medicine, Structural Biology, Nuclear retention, P-bodies, Genetics, MRNA transport, Animals, Nuclear Matrix, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Northern blot, RNA, Messenger, Nuclear protein, Nuclear mRNA metabolism, Molecular Biology, 030304 developmental biology, Cell Nucleus, 0303 health sciences, Messenger RNA, Biological Transport, Cell Biology, Nuclear matrix, Blotting, Northern, Molecular biology, Cell biology, Rats, Kinetics, RNA splicing, DNA Probes, 030217 neurology & neurosurgery, Cell Division
Abstract

International audience; Little is known about the nuclear mRNA content of mammalian cells. In this study, we analyzed by Northern blotting with a panel of probes the nuclear and cytoplasmic fractions derived from several rodent cell lines. For most of the genes under study, mature mRNAs could easily be detected in the nuclear fraction and accumulated to higher levels than the corresponding precursors. In addition, significant differences in the nucleo-cytoplasmic partition of mature mRNAs were observed between genes as well as between cell types (NIH 3T3, CTLL-2, D3-ES, PC-12), indicating that this nuclear accumulation of mRNA is regulated. Thus, while it is usually considered that splicing is the limiting step of pre-mRNA processing, these results point towards transport or nuclear retention of mRNA as a key determinant of nuclear mRNA metabolism.

Published
1999
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29. Novel microarray-based method for estimating exposure to ionizing radiation.

Author

Gruel G, Lucchesi C, Pawlik A, Frouin V, Alibert O, Kortulewski T, Zarour A, Jacquelin B, Gidrol X, and Tronik-Le Roux D

Subjects
Animals, Body Burden, Dose-Response Relationship, Radiation, Male, Mice, Mice, Inbred C57BL, Radiation Dosage, Radiation, Ionizing, Relative Biological Effectiveness, Algorithms, Blood Proteins analysis, Environmental Exposure analysis, Gene Expression radiation effects, Oligonucleotide Array Sequence Analysis methods, Radiometry methods
Abstract

Accurate estimation of the dose of ionizing radiation to which individuals have been exposed is critical for therapeutic treatment. We investigated whether gene expression profiles could be used to evaluate the dose received, thereby serving as a biological dosimeter. We used cDNA microarrays to monitor changes in gene expression profiles induced by ionizing radiation in mouse total blood. The subsets of genes best characterizing each dose were identified by resampling the original data set and calculating the intersection of the dose signatures. This analytical strategy minimizes the impact of potential genetic/epigenetic variation between mice and overcomes the bias in gene selection inherent to microarray technology. The significance of the identified signatures was evaluated by monitoring the type I error rate by in silico negative control simulation. Based on the distribution of the mean ratios of the selected probes, we were able to identify transcription profiles giving 83% to 100% correct estimation of the dose received by test mice, demonstrating that the selected probes could be used to determine the dose of radiation to which the animals had been exposed. This method could potentially be generalized to determine the level of exposure to other toxins and could be used to develop new related clinical applications.

Published
2006
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30. Comparison of gene expression pattern in SP cell populations from four tissues to define common "stemness functions".

Author

Rochon C, Frouin V, Bortoli S, Giraud-Triboult K, Duverger V, Vaigot P, Petat C, Fouchet P, Lassalle B, Alibert O, Gidrol X, and Piétu G

Subjects
Animals, Bone Marrow Cells cytology, Cell Line, Cell Separation, Cells, Cultured, Germinal Center cytology, Male, Mesoderm cytology, Mice, Mice, Inbred C57BL, Muscle Cells cytology, Stem Cells cytology, Bone Marrow Cells classification, Bone Marrow Cells metabolism, Gene Expression Profiling, Germinal Center metabolism, Mesoderm metabolism, Muscle Cells metabolism, Stem Cells metabolism
Abstract

The goal of our study was to identify a subset of genes commonly expressed in Side Populations (SP), isolated by Hoechst staining followed by flow cytometry, from adult mouse bone marrow, male adult germinal cells, muscle primary culture, and mesenchymal cells. These SP cells have been proposed to be a "stem-like" population and are used here as a "model" that may reveal mechanisms which would be relevant for a better understanding of stem cell properties. Transcriptional profiles for SP and the more differentiated non-SP cells isolated from the four tissues were compared by hybridization on microarray using a common external reference. Among the 503 genes differentially expressed, which discriminate SP and non-SP cells in all the tissues, the genes upregulated in SP cells are implicated in the quiescent status of the cells, the maintenance of their pluripotency and the capacity to undergo asymmetric division. These genes may be responsible for the decision for self-renewal of these cells, whereas the repression of lineage-affiliated genes in SP cells could be responsible for their undifferentiated state. These genes, acting in concert, may be the key players that mediate the mechanisms that control stem cell functions, and our results suggest that we have identified common "stemness functions" of these "stem-like" cells.

Published
2006
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31. Cell microarrays in drug discovery.

Author

Castel D, Pitaval A, Debily MA, and Gidrol X

Subjects
Animals, Cell Adhesion, Cells, Cultured metabolism, Extracellular Matrix Proteins metabolism, Gene Expression, Humans, RNA Interference, RNA, Small Interfering genetics, RNA, Small Interfering metabolism, Reproducibility of Results, Drug Design, Oligonucleotide Array Sequence Analysis instrumentation, Oligonucleotide Array Sequence Analysis methods, Protein Array Analysis methods, Tissue Array Analysis methods
Abstract

There is an increasing need for systematic cell-based assays in a high-throughput screening (HTS) format to analyze the phenotypic consequences of perturbing mammalian cells with drugs, genes, interfering RNA. Taking advantage of the recent progress in microtechnology, new cell microarrays are being developed and applied to a large range of issues in metazoan cells. This article compares different approaches and evaluates their potential use in the drug discovery process. Although still an emerging technology, cell microarrays hold great promise to optimize the efficiency:cost ratio in cell-based HTS.

Published
2006
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32. Human side population keratinocytes exhibit long-term proliferative potential and a specific gene expression profile and can form a pluristratified epidermis.

Author

Larderet G, Fortunel NO, Vaigot P, Cegalerba M, Maltère P, Zobiri O, Gidrol X, Waksman G, and Martin MT

Subjects
Adult, Cell Differentiation, Cell Proliferation, Cell Separation, Cells, Cultured, Colony-Forming Units Assay, Female, Gene Expression Profiling, Humans, In Vitro Techniques, Keratinocytes classification, Oligonucleotide Array Sequence Analysis, Stem Cells classification, Time Factors, Epidermal Cells, Epidermis metabolism, Keratinocytes cytology, Keratinocytes metabolism, Stem Cells cytology, Stem Cells metabolism
Abstract

The aim of the present study was to characterize human side population (SP) epidermal keratinocytes isolated from primary cell cultures. For that purpose, keratinocytes were isolated from normal adult breast skin samples and the Hoechst 33342 exclusion assay described for hematopoietic cells was adapted to keratinocytes. Three types of keratinocytes were studied: the SP, the main population (MP), and the unsorted initial population. SP keratinocytes represented 0.16% of the total population. In short-term cultures, they exhibited an increased colony-forming efficiency and produced more actively growing colonies than did unsorted and MP keratinocytes. In long-term cultures, SP cells exhibited an extensive expansion potential, performing a mean of 44 population doublings for up to 12 successive passages after cell sorting. Moreover, even in long-term cultures, SP keratinocytes were able to form a pluristratified epidermis when seeded on a dermal substrate. Unsorted and MP keratinocytes promoted a reduced expansion: mean values of 14 population doublings for five passages and 12 population doublings for four successive passages, respectively. To further characterize SP cells, cDNA microarrays were used to identify their molecular signature. Transcriptome profiling showed that 41 genes were differentially expressed in SP (vs. MP) cells, with 37 upregulated genes and only four downregulated genes in SP cells. The majority of these genes were functionally related to the regulation of transcription and cell signaling. In conclusion, SP human keratinocytes isolated from primary cultures exhibited both short- and long-term high proliferative potential, formed a pluristratified epidermis, and were characterized by a specific gene expression profile.

Published
2006
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33. Global transcriptional characterization of SP and MP cells from the myogenic C2C12 cell line: effect of FGF6.

Author

Decraene C, Benchaouir R, Dillies MA, Israeli D, Bortoli S, Rochon C, Rameau P, Pitaval A, Tronik-Le Roux D, Danos O, Gidrol X, Garcia L, and Piétu G

Subjects
Animals, Benzimidazoles, Cell Differentiation, Cell Separation, Cells, Cultured, DNA metabolism, DNA Probes, Down-Regulation genetics, Flow Cytometry, Gene Expression Profiling, Mice, Microarray Analysis, Principal Component Analysis, Reproducibility of Results, Reverse Transcriptase Polymerase Chain Reaction, Staining and Labeling, Up-Regulation genetics, Fibroblast Growth Factor 6 genetics, Fibroblast Growth Factor 6 metabolism, Myoblasts metabolism, Proto-Oncogene Proteins genetics, Proto-Oncogene Proteins metabolism, Transcription, Genetic genetics
Abstract

With the use of Hoechst staining techniques, we have previously shown that the C2C12 myogenic cell line contains a side population (SP) that is largely increased in the presence of fibroblast growth factor 6 (FGF6). Here, we compared transcriptional profiles from SP and main population (MP) cells from either C2C12 or FGF6-expressing C2C12. Expression profiles of SPs show that these cells are less differentiated than MPs and display some similarities to stem cells. Moreover, principal component analysis made it possible to distinguish specific contributions of either FGF6 or differentiation effects on gene expression profiles. This demonstrated that FGF6-expanded SPs were similar to parental C2C12-derived SPs. Conversely, FGF6-treated MPs differed from parental MPs and were more related to SP cells. These results show that FGF6 pushed committed myogenic cells toward a more immature phenotype resulting in the accumulation of cells with a SP phenotype. We propose that FGF6 conditioning could provide a way to expand the pool of immature cells by myoblast dedifferentiation.

Published
2005
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34. Novel pathway for megakaryocyte production after in vivo conditional eradication of integrin alphaIIb-expressing cells.

Author

Jacquelin B, Kortulewski T, Vaigot P, Pawlik A, Gruel G, Alibert O, Soularue P, Joubert C, Gidrol X, and Tronik-Le Roux D

Subjects
Animals, Cell Lineage, DNA Nucleotidylexotransferase genetics, DNA-Binding Proteins genetics, Enhancer of Zeste Homolog 2 Protein, Gene Expression Profiling, Histone-Lysine N-Methyltransferase, Lymphocytes physiology, Mice, Mice, Transgenic, Myeloid Cells physiology, Polycomb Repressive Complex 2, Proteins, Regeneration, Bone Marrow Cells physiology, Hematopoiesis, Megakaryocytes cytology, Platelet Membrane Glycoprotein IIb genetics
Abstract

Our knowledge of the molecular mechanisms that regulate hematopoiesis in physiologic and pathologic conditions is limited. Using a molecular approach based on cDNA microarrays, we demonstrated the emergence of an alternative pathway for mature bone marrow cell recovery after the programmed and reversible eradication of CD41+ cells in transgenic mice expressing a conditional toxigene targeted by the platelet alphaIIb promoter. The expression profile of the newly produced CD41+ cells showed high levels of transcripts encoding Ezh2, TdT, Rag2, and various immunoglobulin (Ig) heavy chains. In this context, we identified and characterized a novel population of Lin-Sca-1hi c-Kit- cells, with a lymphoid-like expression pattern, potentially involved in the reconstitution process. Our study revealed novel transcriptional cross talk between myeloid and lymphoid lineages and identified gene expression modifications that occur in vivo under these particular stress conditions, opening important prospects for therapeutic applications.

Published
2005
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35. CD98, a novel marker of transient amplifying human keratinocytes.

Author

Lemaître G, Gonnet F, Vaigot P, Gidrol X, Martin MT, Tortajada J, and Waksman G

Subjects
Caveolin 1 metabolism, Cell Line, Electrophoresis, Polyacrylamide Gel, Flow Cytometry, Fusion Regulatory Protein-1 chemistry, Humans, Immunohistochemistry, Immunoprecipitation, Keratinocytes metabolism, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Fusion Regulatory Protein-1 analysis, Keratinocytes chemistry
Abstract

Identification of plasma membrane markers of basal keratinocytes is essential for sorting basal cells and, subsequently, adult epidermal stem cells. In this study, we isolated caveolin-1-enriched microdomains from human HaCaT keratinocytes and identified proteins representing potential cell surface markers of the epidermis by a proteomic approach. The purification of this caveolae domain allowed us to characterize 53 proteins of which 26% were transmembrane and 32% associated-membrane proteins. One of them, CD98, was found to be co-localized with beta1 integrin at the plasma membrane of the basal keratinocytes of healthy human epidermis. We then isolated CD98-positive keratinocytes from fresh skin biopsies. Using clonogenic assays, we demonstrate that CD98 may be considered as a marker of transient amplifying human keratinocytes.

Published
2005
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36. Activation of an energy providing response in human keratinocytes after gamma irradiation.

Author

Lamartine J, Franco N, Le Minter P, Soularue P, Alibert O, Leplat JJ, Gidrol X, Waksman G, and Martin MT

Subjects
Cells, Cultured, Energy Metabolism genetics, Humans, Keratinocytes cytology, Energy Metabolism radiation effects, Gamma Rays, Keratinocytes metabolism, Mitochondria enzymology
Abstract

We performed a microarray study on human differentiated HaCaT keratinocytes exposed to ionizing radiation (2 or 10 Gy). At 3 h after exposure, more than 150 known and unknown genes were found regulated in irradiated HaCaT keratinocytes. Among the genes regulated at 3 h, those involved in cell energy metabolism appeared to be the most abundant and the most responsive. Two mitochondrial ATP-synthases and several other genes involved in energy producing pathways, such as glucose metabolism, were induced, whereas many genes from energy requiring pathways were shut down. These changes in energy metabolism were confirmed both in normal primary keratinocytes and in HaCaT keratinocytes by RT-PCR and proteins studies. Moreover, measures of intracellular ATP revealed a 50% increase in keratinocytes immediately after irradiation, supporting an energy procurement response. The overall results indicate that irradiation induces an immediate burst of ATP that seems to be a general response of human differentiated keratinocytes to the radiation stress. This article contains Supplementary Material available at http://www.mrw.interscience.wiley.com/suppmat/0730-2312/suppmat/v95.html, ((c) 2005 Wiley-Liss, Inc.)

Published
2005
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37. Low-dose exposure to gamma rays induces specific gene regulations in normal human keratinocytes.

Author

Franco N, Lamartine J, Frouin V, Le Minter P, Petat C, Leplat JJ, Libert F, Gidrol X, and Martin MT

Subjects
Cells, Cultured, Dose-Response Relationship, Radiation, Humans, Radiation Dosage, Skin metabolism, Skin radiation effects, Gamma Rays, Gene Expression Regulation physiology, Gene Expression Regulation radiation effects, Keratinocytes metabolism, Keratinocytes radiation effects, Transcription Factors metabolism
Abstract

Skin is the organ most exposed to various environmental aggressors, including ionizing radiation. Low-dose and low-dose-rate exposures to gamma rays account for most occupational, medical or environmental irradiations. To examine whether this type of exposure triggers specific molecular responses, cultured primary keratinocytes isolated from adult normal skin were irradiated with single acute doses of 1 cGy or 2 Gy. DNA microarrays containing 10,500 probes were used to assess transcriptional changes over a time course between 3 and 72 h postirradiation. Keratinocytes were studied at a differentiated stage to mimic the response of cells from the suprabasal layers of the epidermis. A major finding of this study was the identification of an important number of low-dose-specific genes (140), most of which were modulated at 48 h. Clustering analysis also revealed low-dose-specific profiles. One of these clusters (17 known genes) was further analyzed using Gibbs sampling algorithm, which led to the identification of 7 putative promoter sequences. These results show for the first time that low-dose ionizing radiation is able to induce specific transcriptional responses in human keratinocytes. Our findings support the potential usefulness of microarrays in biological dosimetry studies after low-dose exposures.

Published
2005
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38. [Cell microarrays and functional genomics].

Author

Roupioz Y, Castel D, Pitaval A, Baghdoyan S, and Gidrol X

Subjects
Cells, Cultured, Humans, Transfection methods, Genomics methods, Oligonucleotide Array Sequence Analysis methods
Abstract

With the complete sequencing of the human genome, research priorities have shifted from the identification of genes to the elucidation of their function. Methods currently used by scientists to characterize gene function, such as knock-out mice, are based upon loss of protein function and analysis of the resulting phenotypes to infer a potential role for the protein under scrutiny. Until now, these methods have been successful but time consuming and only a few genes at a time could be analyzed. Cell microarrays allow to simultaneously transfect thousands of different nucleic acid molecules, RNA or DNA, into adherent cells. It is then possible to analyze a large pallet of resulting phenotypes in clusters of transfected cells. We are currently manufacturing cell microarrays with collections of full-length cDNA cloned in expression vectors (gain of function analyses) or siRNA (loss of function studies) to unravel function of genes involved in differentiation and proliferation of human cells. Although there are still some technological difficulties to overcome, the potential for cell microarrays to speed up functional exploration of genomes is very promising.

Published
2005
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39. Gene structure and promoter analysis of the human GJB6 gene encoding connexin 30.

Author

Essenfelder GM, Larderet G, Waksman G, and Lamartine J

Subjects
5' Flanking Region genetics, Alternative Splicing, Base Sequence, Cell Line, Connexin 30, Connexins analysis, DNA chemistry, DNA genetics, Epidermal Cells, Epidermal Growth Factor pharmacology, Epidermis metabolism, Gene Expression drug effects, Genes genetics, Hair Follicle chemistry, Humans, Immunohistochemistry, Keratinocytes metabolism, Luciferases genetics, Luciferases metabolism, Molecular Sequence Data, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins metabolism, Regulatory Sequences, Nucleic Acid genetics, Sequence Alignment, Sequence Analysis, DNA, Transcription, Genetic, Transfection, Connexins genetics, Promoter Regions, Genetic genetics
Abstract

Connexins (Cx) are the protein subunits of gap junctions, which play an important role in cell-to-cell communication. We characterized the genomic structure of the human GJB6 gene, encoding connexin 30 (C x 30), and showed that it differs from most connexin-encoding genes. GJB6 presents six different exons, some of which can be alternatively spliced. We also mapped a basal promoter sequence active in a human keratinocyte cell line which responds to the activation of the EGF receptor. One of the non-encoding exons of GJB6, which has been described in brain C x 30 cDNA, was not found in cDNA obtained from human keratinocytes, suggesting tissue-specific splicing.

Published
2005
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40. Id2 reverses cell cycle arrest induced by {gamma}-irradiation in human HaCaT keratinocytes.

Author

Baghdoyan S, Lamartine J, Castel D, Pitaval A, Roupioz Y, Franco N, Duarte M, Martin MT, and Gidrol X

Subjects
Cell Cycle physiology, Cell Proliferation radiation effects, Humans, Inhibitor of Differentiation Protein 2, Keratinocytes cytology, Keratinocytes metabolism, Cell Cycle radiation effects, DNA-Binding Proteins metabolism, Gamma Rays, Keratinocytes radiation effects, Repressor Proteins metabolism, Transcription Factors metabolism
Abstract

Id2 plays a key role in epithelial cells, regulating differentiation, the cell cycle, and proliferation. Because human skin constantly renews itself and is the first target of irradiation, it is of primary interest to evaluate whether such a gene may be regulated in keratinocytes exposed to ionizing radiation. We show here that Id2 is induced in response to gamma-irradiation and have investigated the consequence of this regulation on cell fate. Using RNA interference, we observed that Id2 extinction significantly reduces cell growth in human keratinocytes through the control of the G(1)-S transition of the cell cycle. We have investigated whether the impact of Id2 on the cell cycle may have a physiological role on the cell's ability to cope with radiative stress. Indeed, when Id2 is down-regulated through interfering RNA, cells are more sensitive to irradiation. Conversely, when Id2 is overexpressed, this somehow protects the cell. We propose that Id2 favors reentering the cell cycle after radiation-induced cell cycle arrest to permit the recovery of keratinocytes exposed to ionizing radiation.

Published
2005
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41. [Therapeutic effect of human mesenchymal stem cells in skin after radiation damage].

Author

Bensidhoum M, Gobin S, Chapel A, Lemaitre G, Bouet S, Waksman G, Thierry D, and Martin MT

Subjects
Animals, Biomarkers, Bone Marrow Cells, Cell Lineage, Cell Movement, Cells, Cultured transplantation, Graft Survival, Humans, Hyperplasia, Mice, Mice, Inbred NOD, Mice, SCID, Radiation Injuries, Experimental pathology, Radiodermatitis pathology, Transplantation, Heterologous, beta 2-Microglobulin analysis, Mesenchymal Stem Cell Transplantation, Radiation Injuries, Experimental surgery, Radiodermatitis surgery
Abstract

Over 50% of all cancer patients presently receive radiotherapy at one stage in their treatment course. Inevitably skin is one of the most frequently damaged tissue due to its localization and constant turn-over. Our present goal is to reduce radiation-induced complications in human skin through stem cell therapy, particulary in human epidermis. Mesenchymal Stem Cells (MSCs) have been shown to be multipotent cells able to engraft in many tissues after injury. Herein, we isolated human MSCs and tested their capability to improve skin wound healing after irradiation. This potential was assessed in NOD/SCID mice which received 30 Gy locally on the thigh. This dose caused within 3 weeks local epidermis necrosis which was repaired within 13 weeks. MSCs were intravenously injected in irradiated mice 24 hours after exposure. Clinical scoring throughout 6 weeks gave indications that human MSCs reduced the extent of damage and accelerated the wound healing process. We show by quantitative qPCR and histological studies the presence of human MSCs derived cells into the scar. Human MSCs homed to the damaged skin and participated to the wound healing process. These results open prospects for cellular therapy by MSCs in irradiated epithelial tissues and could be extended to the whole general field of cutaneous cicatrization, particularly after burns.

Published
2005
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42. Expression profiling of genes and proteins in HaCaT keratinocytes: proliferating versus differentiated state.

Author

Lemaître G, Lamartine J, Pitaval A, Vaigot P, Garin J, Bouet S, Petat C, Soularue P, Gidrol X, Martin MT, and Waksman G

Subjects
Animals, Cell Cycle physiology, Cell Line, Humans, Keratinocytes cytology, Keratinocytes metabolism, Molecular Sequence Data, Oligonucleotide Array Sequence Analysis, Signal Transduction physiology, Cell Differentiation physiology, Cell Proliferation, Gene Expression Profiling, Gene Expression Regulation, Keratinocytes physiology
Abstract

The knowledge of the mechanism of keratinocyte differentiation in culture is still uncompleted. The emergence of new technologies, such as cDNA microarrays or 2D electrophoresis followed by mass spectrometry analysis, has allowed the identification of genes and proteins expressed in biological processes in keratinocytes. Here, we report a genome wide analysis of proliferating versus differentiated human HaCaT keratinocytes. We found that genes and proteins which take part in the cell cycle control, carbohydrate metabolism, cell auto-immunity, adhesion and cytokine signal transduction pathways were regulated in differentiated HaCaT keratinocytes. In addition, we identified seven proteins and 33 transcripts that had not been previously described as differentially expressed in proliferating versus differentiated HaCaT cells. Furthermore, some of these transcripts or proteins were similarly regulated in human primary keratinocytes and in human epidermis. The present study opens new areas of investigation in the comprehension of keratinocyte differentiation., ((c) 2004 Wiley-Liss, Inc.)

Published
2004
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43. Connexin30 mutations responsible for hidrotic ectodermal dysplasia cause abnormal hemichannel activity.

Author

Essenfelder GM, Bruzzone R, Lamartine J, Charollais A, Blanchet-Bardon C, Barbe MT, Meda P, and Waksman G

Subjects
Adenosine Triphosphate metabolism, Animals, Connexin 30, DNA Primers, Electrophysiology, Epidermis metabolism, HeLa Cells, Humans, Immunohistochemistry, Ion Channels genetics, Microinjections, Mutagenesis, Site-Directed, Nucleic Acid Amplification Techniques, Oocytes metabolism, Transfection, Xenopus, Connexins genetics, Connexins metabolism, Ectodermal Dysplasia genetics, Ion Channels metabolism, Mutation genetics
Abstract

Clouston syndrome or hidrotic ectodermal dysplasia (HED) is a rare dominant genodermatosis characterized by palmoplantar hyperkeratosis, generalized alopecia and nail defects. The disease is caused by mutations in the human GJB6 gene which encodes the gap junction protein connexin30 (Cx30). To gain insight into the molecular mechanisms underlying HED, we have analyzed the consequences of two of these mutations (G11R Cx30 and A88V Cx30) on the functional properties of the connexons they form. Here, we show that the distribution of Cx30 is similar in affected palmoplantar skin and in normal epidermis. We further demonstrate that the presence of the wild-type protein (wt Cx30) improves the trafficking of mutated Cx30 to the plasma membrane where both G11R and A88V Cx30 co-localize with wt Cx30 and form functional intercellular channels. The electrophysiological properties of channels made of G11R and A88V Cx30 differ slightly from those of wt Cx30 but allow for dye transfer between transfected HeLa cells. Finally, we document a gain of function of G11R and A88V Cx30, which form functional hemichannels at the cell surface and, when expressed in HeLa cells, generate a leakage of ATP into the extracellular medium. Such increased ATP levels might act as a paracrine messenger that, by altering the epidermal factors which control the proliferation and differentiation of keratinocytes, may play an important role in the pathophysiological processes leading to the HED phenotype.

Published
2004
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44. Quantitative analysis of highly parallel transfection in cell microarrays.

Author

Baghdoyan S, Roupioz Y, Pitaval A, Castel D, Khomyakova E, Papine A, Soussaline F, and Gidrol X

Subjects
Automation, Biopolymers, Cell Line, Computational Biology methods, DNA administration & dosage, DNA genetics, Fluorescence, Gelatin, Gene Expression Profiling methods, Gene Silencing, Genes, Reporter genetics, Genetic Vectors genetics, Genome, Human, Green Fluorescent Proteins, Humans, Luminescent Proteins genetics, Phenotype, RNA, Small Interfering administration & dosage, RNA, Small Interfering genetics, RNA, Small Interfering metabolism, Reproducibility of Results, Software, Cells metabolism, Genomics methods, Oligonucleotide Array Sequence Analysis methods, Transfection methods, Transfection standards
Abstract

As more genomes are sequenced, we are facing the challenge of rapidly unraveling the functions of genes. To that end, cell microarrays have recently been described that transfect thousands of nucleic acids in parallel and can be used to analyze the phenotypic consequences of such perturbations. As many parameters can influence the efficacy of transfection in such a format, we describe some important features in manufacturing cell microarrays that may improve reliability and efficiency of both plasmid DNA and siRNA transfection. We have also developed image analysis software that allows automatic detection of cell clusters, quantification of transfection efficiency and levels of expression/extinction of genes. Along with cell microarrays, this bioinformatic tool should expedite functional exploration of the human genome.

Published
2004
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45. Altered proliferation and differentiation of human epidermis in cases of skin fibrosis after radiotherapy.

Author

Sivan V, Vozenin-Brotons MC, Tricaud Y, Lefaix JL, Cosset JM, Dubray B, and Martin MT

Subjects
Adult, Aged, Cell Differentiation radiation effects, Cell Division radiation effects, Cicatrix metabolism, Female, Humans, Hyperplasia etiology, Hyperplasia metabolism, Hyperplasia pathology, Integrins metabolism, Keratinocytes metabolism, Keratinocytes pathology, Keratins metabolism, Male, Middle Aged, Proliferating Cell Nuclear Antigen metabolism, Radiation Injuries metabolism, Skin metabolism, Skin pathology, Transforming Growth Factor beta metabolism, Cicatrix pathology, Keratinocytes radiation effects, Radiation Injuries pathology, Skin radiation effects
Abstract

Purpose: To characterize, at the histopathologic and molecular levels, the irradiated epidermis in cases of human skin fibrosis induced by radiotherapy., Methods and Materials: Surgical samples were obtained from 6 patients who had developed cutaneous fibronecrotic lesions from 7 months to 27 years after irradiation. The proliferation and differentiation status of the irradiated epidermis was characterized with specific markers using immunohistochemical methods., Results: All samples presented with hyperplasia of the epidermis associated with local inflammation. The scar epidermis exhibited an increased expression of proliferating cell nuclear antigen, which revealed hyperproliferation of keratinocytes. Furthermore, an abnormal differentiation was found, characterized by the expression of K6 and K16, and by alterations in protein amounts and localization of cytokeratins, involucrin, and transforming growth factor-beta1., Conclusion: These results demonstrate that late damage of irradiated skin is not only characterized by fibrosis in the dermis but also by hyperplasia in the epidermis. This hyperplasia was due to both hyperproliferation and abnormal differentiation of keratinocytes.

Published
2002
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46. Mutations in GJB6 cause hidrotic ectodermal dysplasia.

Author

Lamartine J, Munhoz Essenfelder G, Kibar Z, Lanneluc I, Callouet E, Laoudj D, Lemaître G, Hand C, Hayflick SJ, Zonana J, Antonarakis S, Radhakrishna U, Kelsell DP, Christianson AL, Pitaval A, Der Kaloustian V, Fraser C, Blanchet-Bardon C, Rouleau GA, and Waksman G

Subjects
Amino Acid Sequence, Amino Acid Substitution, Animals, Arginine, Chromosome Mapping, Connexin 30, Female, Glycine, Humans, Male, Mice, Molecular Sequence Data, Pedigree, Sequence Alignment, Sequence Homology, Amino Acid, Chromosomes, Human, Pair 13, Connexins genetics, Ectodermal Dysplasia genetics, Mutation, Missense
Published
2000
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