Descriptor: "MESH : Immunohistochemistry" - Searchworks@Jio Institute Digital Library Search Results

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46 results on '"MESH : Immunohistochemistry"'

1. Adrenal Cortex Remodeling and Functional Zona Glomerulosa Hyperplasia in Primary Aldosteronism

Author: Arndt Benecke, Xavier Jeunemaitre, Estelle Louiset, José-Felipe Golib Dzib, Pierre-François Plouin, Sheerazed Boulkroun, Tchao Meatchi, Enzo Lalli, Laurence Amar, Benoit Samson-Couterie, Hervé Lefebvre, Maria-Christina Zennaro, Paris-Centre de Recherche Cardiovasculaire (PARCC - UMR-S U970), Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Paris Descartes - Paris 5 (UPD5), Institut des Hautes Etudes Scientifiques (IHES), IHES, Différenciation et communication neuronale et neuroendocrine (DC2N), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), UNIROUEN - UFR Santé (UNIROUEN UFR Santé), Normandie Université (NU)-Normandie Université (NU), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO), Institut de pharmacologie moléculaire et cellulaire (IPMC), Centre National de la Recherche Scientifique (CNRS)-Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA), Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA), Institut de Recherche Interdisciplinaire [Villeneuve d'Ascq] (IRI), Centre National de la Recherche Scientifique (CNRS)-Université de Lille, Droit et Santé-Université de Lille, Sciences et Technologies, Service de génétique [CHU HEGP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO), Paris-Centre de Recherche Cardiovasculaire ( PARCC - U970 ), Hôpital Européen Georges Pompidou [APHP] ( HEGP ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université Paris Descartes - Paris 5 ( UPD5 ), Institut des Hautes Etudes Scientifiques ( IHES ), Différenciation et communication neuronale et neuroendocrine ( DC2N ), Université de Rouen Normandie ( UNIROUEN ), Normandie Université ( NU ) -Normandie Université ( NU ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), UNIROUEN - UFR Santé, Normandie Université ( NU ) -Normandie Université ( NU ), Hôpital Européen Georges Pompidou [APHP] ( HEGP ), Institut de pharmacologie moléculaire et cellulaire ( IPMC ), Université Nice Sophia Antipolis ( UNS ), Université Côte d'Azur ( UCA ) -Université Côte d'Azur ( UCA ) -Centre National de la Recherche Scientifique ( CNRS ), Université Côte d'Azur ( UCA ), Institut de Recherche Interdisciplinaire [Villeneuve d'Ascq] ( IRI ), Université de Lille, Sciences et Technologies-Université de Lille, Droit et Santé-Centre National de la Recherche Scientifique ( CNRS ), Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Européen Georges Pompidou [APHP] ( HEGP ), Université Paris Descartes - Paris 5 (UPD5)-Hôpital Européen Georges Pompidou [APHP] (HEGP), Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP), Université Côte d'Azur (UCA)-Université Côte d'Azur (UCA)-Centre National de la Recherche Scientifique (CNRS), Université Côte d'Azur (UCA), Université de Lille, Sciences et Technologies-Université de Lille, Droit et Santé-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Européen Georges Pompidou [APHP] (HEGP), and Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)
Subjects: Male, MESH : Steroid 11-beta-Hydroxylase, MESH : Immunohistochemistry, MESH : Adrenal Cortex Neoplasms, MESH: Adrenal Cortex Neoplasms, MESH : Adrenal Cortex, chemistry.chemical_compound, 0302 clinical medicine, Primary aldosteronism, MESH : Female, Aldosterone, In Situ Hybridization, 0303 health sciences, MESH: Middle Aged, Adrenal cortex, MESH : In Situ Hybridization, Middle Aged, MESH : Adult, Hyperplasia, Immunohistochemistry, MESH : Hyperaldosteronism, MESH: Hyperaldosteronism, medicine.anatomical_structure, Female, Zona Glomerulosa, Adenoma, Adult, medicine.medical_specialty, MESH: Zona Glomerulosa, medicine.drug_class, MESH : Male, 030209 endocrinology & metabolism, Biology, MESH: Aldosterone Synthase, 03 medical and health sciences, MESH: In Situ Hybridization, MESH : Hyperplasia, [ SDV.MHEP ] Life Sciences [q-bio]/Human health and pathology, Internal medicine, Hyperaldosteronism, Internal Medicine, medicine, Cytochrome P-450 CYP11B2, Humans, MESH : Middle Aged, Steroid 11-beta-hydroxylase, 030304 developmental biology, MESH : Aldosterone Synthase, MESH: Adenoma, MESH: Humans, MESH : Aldosterone, MESH: Hyperplasia, MESH : Humans, MESH: Aldosterone, MESH: Immunohistochemistry, MESH: Adult, medicine.disease, Adrenal Cortex Neoplasms, MESH: Male, Endocrinology, MESH : Adenoma, chemistry, Mineralocorticoid, Zona glomerulosa, Adrenal Cortex, Steroid 11-beta-Hydroxylase, MESH: Adrenal Cortex, MESH : Zona Glomerulosa, MESH: Female, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology, MESH: Steroid 11-beta-Hydroxylase
Abstract: International audience; Primary aldosteronism is the most common form of secondary hypertension with hypokalemia and suppressed renin-angiotensin system caused by autonomous aldosterone production. Our aim was to compare zona glomerulosa (ZG) structure and function between control adrenals and the peritumoral tissue from patients operated on for aldosterone-producing adenoma. ZG morphology and CYP11B1, CYP11B2, and disabled 2 expression were studied in 15 control adrenals and 25 adrenals with aldosterone-producing adenoma. A transcriptome analysis was done using publicly available data sets. In control adrenals, ZG was discontinuous, and CYP11B2 expression was focal or partly continuous and localized to 3 structures, foci, megafoci, and aldosterone-producing cell clusters. CYP11B2 expression was restricted to a limited number of ZG cells expressing Dab2 but not CYP11B1; aldosterone-producing cell clusters were composed of cells with an intermediate phenotype expressing CYP11B2 but not disabled 2 or CYP11B1. In peritumoral tissue, large remodeling of the adrenal cortex was observed with increased nodulation and decreased vascularization that were not correlated with CYP11B2 expression. In 17 out of 25 adrenals, hyperplasia of adjacent ZG was observed with persistent expression of CYP11B2 that was extended to the entire ZG. In all of the adrenals from patients with aldosterone-producing adenoma, CYP11B2 expression was present in foci, megafoci, and aldosterone-producing cell clusters. Transcriptome profiling indicates a close relationship between peritumoral and control adrenal cortex. In conclusion, adrenal cortex remodeling, reduced vascularization, and ZG hyperplasia are major features of adrenals with aldosterone-producing adenoma. Transcriptional phenotyping is not in favor of this being an intermediate step toward the formation of aldosterone-producing adenoma.
Published: 2010

2. Hypoxia Down-regulates CCAAT/Enhancer Binding Protein-α Expression in Breast Cancer Cells

Author: Robert Barouki, Etienne Blanc, Nathalie M. Mazure, Fabrice Lecuru, Marie-Claude Fulchignoni-Lataud, Marie-Aude Le Frere Belda, Anne Dreiem, Thérèse Hervèe Mayi, Charbel Massaad, Vincent Favaudon, Liliane Massaad-Massade, Ramzi Seifeddine, Laboratoire de Détection et de Géophysique (CEA) (LDG), DAM Île-de-France (DAM/DIF), Direction des Applications Militaires (DAM), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction des Applications Militaires (DAM), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO), IFR50, Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Faculté de Médecine Nice, Institut de signalisation, biologie du développement et cancer (ISBDC), Centre National de la Recherche Scientifique (CNRS)-Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA), Génotoxicologie, signalisation et radiothérapie expérimentale, Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Biophysique moléculaire, Régulation de la transcription et maladies génétiques (RTMG), Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Stéroïdes et système nerveux : physiopathologie moléculaire et clinique, Institut National de la Santé et de la Recherche Médicale (INSERM), Faculté de Médecine Paris-Sud, Université Paris-Sud - Paris 11 (UP11), Toxicologie Moleculaire (U490), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), 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)-Université Côte d'Azur (UCA), Laboratoire de Détection et de Géophysique (CEA) ( LDG ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), Hôpital Européen Georges Pompidou [APHP] ( HEGP ), Université Nice Sophia Antipolis ( UNS ), Université Côte d'Azur ( UCA ) -Université Côte d'Azur ( UCA ) -Faculté de Médecine Nice, Institut de signalisation, biologie du développement et cancer ( ISBDC ), Université Côte d'Azur ( UCA ) -Université Côte d'Azur ( UCA ) -Centre National de la Recherche Scientifique ( CNRS ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -INSTITUT CURIE, Régulation de la transcription et maladies génétiques ( RTMG ), Centre National de la Recherche Scientifique ( CNRS ) -Centre National de la Recherche Scientifique ( CNRS ), Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université Paris-Sud - Paris 11 ( UP11 ), Toxicologie Moleculaire, Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP), Université Côte d'Azur (UCA)-Université Côte d'Azur (UCA)-Faculté de Médecine Nice, Université Côte d'Azur (UCA)-Université Côte d'Azur (UCA)-Centre National de la Recherche Scientifique (CNRS), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Curie
Subjects: Cancer Research, Hypoxia-Inducible Factor 1, MESH : Molecular Sequence Data, MESH : RNA, Messenger, Transcription, Genetic, MESH : Immunohistochemistry, MESH: Cell Hypoxia, Electrophoretic Mobility Shift Assay, MESH: Cell Cycle, RNA polymerase II, MESH : Blotting, Western, MESH : Breast Neoplasms, MESH: Base Sequence, MESH : RNA, Small Interfering, MESH: Down-Regulation, MESH : Down-Regulation, 0302 clinical medicine, Transcription (biology), MESH: Reverse Transcriptase Polymerase Chain Reaction, Enhancer binding, MESH: RNA, Small Interfering, Gene expression, MESH: CCAAT-Enhancer-Binding Protein-alpha, MESH : CCAAT-Enhancer-Binding Protein-alpha, RNA, Small Interfering, Promoter Regions, Genetic, 0303 health sciences, Gene knockdown, Reverse Transcriptase Polymerase Chain Reaction, Cell Cycle, MESH : Reverse Transcriptase Polymerase Chain Reaction, Immunohistochemistry, Cell Hypoxia, 3. Good health, MESH: Promoter Regions (Genetics), Oncology, MESH : Electrophoretic Mobility Shift Assay, 030220 oncology & carcinogenesis, MESH: Cell Growth Processes, MESH : Cell Hypoxia, MESH : Transfection, Subcellular Fractions, medicine.medical_specialty, MESH : Hypoxia-Inducible Factor 1, alpha Subunit, MESH: Cell Line, Tumor, Blotting, Western, Molecular Sequence Data, MESH : Deferoxamine, Down-Regulation, Breast Neoplasms, Cell Growth Processes, MESH : Subcellular Fractions, Deferoxamine, Biology, Transfection, MESH: Hypoxia-Inducible Factor 1, alpha Subunit, 03 medical and health sciences, Cell Line, Tumor, Internal medicine, MESH : Cell Cycle, CCAAT-Enhancer-Binding Protein-alpha, medicine, Humans, MESH: Blotting, Western, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, RNA, Messenger, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Transcription factor, MESH: RNA, Messenger, 030304 developmental biology, MESH: Humans, MESH: Molecular Sequence Data, Base Sequence, MESH : Cell Line, Tumor, MESH: Transcription, Genetic, MESH: Transfection, MESH : Humans, MESH : Transcription, Genetic, MESH: Immunohistochemistry, MESH: Deferoxamine, Hypoxia-Inducible Factor 1, alpha Subunit, MESH : Promoter Regions (Genetics), Molecular biology, Endocrinology, HIF1A, MESH : Cell Growth Processes, MESH: Subcellular Fractions, MESH: Electrophoretic Mobility Shift Assay, biology.protein, MESH : Base Sequence, MESH: Breast Neoplasms
Abstract: International audience; The transcription factor CCAAT/enhancer binding protein-alpha (C/EBP alpha) is involved in the control of cell differentiation and proliferation, and has been suggested to act as a tumor suppressor in several cancers. By using microarray analysis, we have previously shown that hypoxia and estrogen down-regulate C/EBP alpha mRNA in T-47D breast cancer cells. Here, we have examined the mechanism by which the down-regulation by hypoxia takes place. Using the specific RNA polymerase II inhibitor 5,6-dichlorobenzimidazole-1-beta-D-ribofuranoside, the mRNA stability was analyzed under normoxia or hypoxia by quantitative reverse transcription-PCR. Hypoxia reduced the half-life of C/EBP alpha mRNA by approximately 30%. C/EBP alpha gene promoter studies indicated that hypoxia also repressed the transcription of the gene and identified a hypoxia-responsive element (-522; -527 bp), which binds to hypoxia-inducible factor (HIF)-1 alpha, as essential for down-regulation of C/EBP alpha transcription in hypoxia. Immunocytochemical analysis showed that C/EBP alpha was localized in the nucleus at 21% O(2), but was mostly cytoplasmic under 1% O(2). Knockdown of HIF-1 alpha by RNAi restored C/EBP alpha to normal levels under hypoxic conditions. Immunohistochemical studies of 10 tumor samples did not show any colocalization of C/EBP alpha and glucose transporter 1 (used as a marker for hypoxia). Taken together, these results show that hypoxia down-regulates C/EBP alpha expression in breast cancer cells by several mechanisms, including transcriptional and posttranscriptional effects. The down-regulation of C/EBP alpha in hypoxia is mediated by HIF-1.
Published: 2008

3. Co-localization of histamine with GABA but not with galanin in the human tuberomamillary nucleus

Author: B. Fisser, C. Chotard, Suzanne Trottier, Dick F. Swaab, J.-C. Schwartz, Unga A. Unmehopa, Elisabeth Traiffort, Netherlands Institute for Neuroscience (NIN), Outters-Lafaye, Michèle, Unité de Neurobiologie et Pharmacologie Moléculaire, Université Paris Descartes - Paris 5 (UPD5) - Institut National de la Santé et de la Recherche Médicale (INSERM) - Centre National de la Recherche Scientifique (CNRS), Netherlands Institute for Brain Research, Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and Other departments
Subjects: Male, MESH : RNA, Messenger, MESH : Immunohistochemistry, MESH : Aged, MESH: gamma-Aminobutyric Acid, Histidine Decarboxylase, chemistry.chemical_compound, MESH: Aged, 80 and over, MESH : Female, MESH : Autopsy, In Situ Hybridization, gamma-Aminobutyric Acid, MESH: Aged, Aged, 80 and over, MESH : Isoenzymes, MESH: Middle Aged, Glutamate Decarboxylase, General Neuroscience, Glutamate receptor, MESH : In Situ Hybridization, MESH : Hypothalamic Area, Lateral, MESH: Hypothalamic Area, Lateral, Middle Aged, MESH : Adult, MESH : Galanin, Immunohistochemistry, MESH : gamma-Aminobutyric Acid, Isoenzymes, MESH : Histamine, medicine.anatomical_structure, Hypothalamus, MESH: Isoenzymes, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Female, Autopsy, MESH: Histamine, Tuberomammillary nucleus, Histamine, MESH: Glutamate Decarboxylase, Adult, medicine.medical_specialty, MESH : Histidine Decarboxylase, MESH : Male, Galanin, In situ hybridization, Biology, MESH: In Situ Hybridization, MESH: Galanin, Internal medicine, medicine, Humans, MESH : Middle Aged, RNA, Messenger, [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH : Aged, 80 and over, Molecular Biology, MESH: RNA, Messenger, Aged, MESH : Glutamate Decarboxylase, MESH: Humans, MESH : Humans, Histaminergic, MESH: Adult, MESH: Immunohistochemistry, MESH: Male, Endocrinology, nervous system, chemistry, Hypothalamic Area, Lateral, Neurology (clinical), MESH: Autopsy, MESH: Female, MESH: Histidine Decarboxylase, Nucleus, Developmental Biology
Abstract: International audience; The presence of GABA and galanin in histaminergic neurons was previously reported in the rodent brain but whether such co-localizations also occur in the human brain was not known. We used in situ hybridization histochemistry and immunohistochemistry to study the co-localization of histamine with GABA and galanin in neurons of the tuberomamillary nucleus of adult human posterior hypothalamus. On consecutive formalin-fixed paraffin-embedded sections, co-localization was assessed using the in situ hybridization for L-histidine decarboxylase mRNA and immunocytochemistry for glutamate decarboxylase-67 kDa or galanin in the two profiles of same cell. The pattern of distribution and number of histaminergic neurons identified by in situ hybridization of the synthesizing enzyme gene transcripts were in accordance with data reported for histamine immunoreactivity. The great majority of neurons within the main divisions of the tuberomamillary nucleus containing L-histidine decarboxylase mRNA was also immunoreactive for glutamate decarboxylase-67 kDa. The range of co-localization of the two markers varied from 72% in the lateral part, to 75-87% in the medial part and 83-88% in the ventral part. In contrast, no cell containing L-histidine decarboxylase mRNA was immunoreactive for galanin. We conclude that tuberomamillary neurons in human co-express histamine with GABA but, unlike the neurons in rodents, do not express galanin, indicating that neurotransmitter co-localization patterns differ in the two species.
Published: 2002

4. Cell-penetrating anti-GFAP VHH and corresponding fluorescent fusion protein VHH-GFP spontaneously cross the blood-brain barrier and specifically recognize astrocytes: application to brain imaging

Author: Jean-Pierre Bourgeois, Anne-Marie Le Sourd, Fabienne Glacial, Pierre Lafaye, Susanna Celli, Pierre-Olivier Couraud, Babette B. Weksler, Ignacio A. Romero, Tengfei Li, Salah Mecheri, François Rougeon, Production de Protéines Recombinantes et d'Anticorps (Plate-Forme), Institut Pasteur [Paris] (IP), Gènes, Synapses et Cognition (CNRS - UMR3571 ), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Biologie des Interactions Hôte-Parasite, Division of Hematology and Oncology, Weill Medical College of Cornell University [New York], Department of Life, Health and Chemical Sciences, The Open University [Milton Keynes] (OU), Génétique et Biochimie du Développement, Institut Pasteur [Paris], Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), 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)-Institut Pasteur [Paris], Gènes, Synapses et Cognition, Institut Pasteur [Paris]-Université Paris Diderot - Paris 7 ( UPD7 ) -Centre National de la Recherche Scientifique ( CNRS ), Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut Cochin ( UM3 (UMR 8104 / U1016) ), 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 Pasteur [Paris]-Centre National de la Recherche Scientifique ( CNRS ), and The Open University [Milton Keynes] ( OU )
Subjects: MESH : Cell Line, [SDV.IB.IMA]Life Sciences [q-bio]/Bioengineering/Imaging, Plasmodium berghei, MESH : Immunohistochemistry, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Fluorescent Antibody Technique, Biochemistry, law.invention, Green fluorescent protein, MESH : Blood-Brain Barrier, MESH: Blood-Brain Barrier, Mice, 0302 clinical medicine, law, MESH: Glial Fibrillary Acidic Protein, MESH : Female, MESH: Animals, MESH: Fluorescent Antibody Technique, [ SDV.IB.IMA ] Life Sciences [q-bio]/Bioengineering/Imaging, 0303 health sciences, biology, Glial fibrillary acidic protein, intrabodies, MESH: Enzyme-Linked Immunosorbent Assay, Brain, MESH : Enzyme-Linked Immunosorbent Assay, MESH : Glial Fibrillary Acidic Protein, Immunohistochemistry, nanobodies, MESH : Mutagenesis, Site-Directed, 3. Good health, MESH: Mutagenesis, Site-Directed, medicine.anatomical_structure, Blood-Brain Barrier, [ SDV.NEU.NB ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH : Single-Domain Antibodies, Recombinant DNA, Female, Antibody, Biotechnology, MESH: Cell Line, Tumor, MESH : Recombinant Fusion Proteins, Recombinant Fusion Proteins, [ SDV.IMM.IMM ] Life Sciences [q-bio]/Immunology/Immunotherapy, fluobodies, Enzyme-Linked Immunosorbent Assay, MESH : Mice, Inbred C57BL, Astrocytoma, Blood–brain barrier, MESH: Single-Domain Antibodies, camelids, Cell Line, 03 medical and health sciences, MESH: Brain, Antigen, MESH: Mice, Inbred C57BL, Cell Line, Tumor, MESH : Mice, Glial Fibrillary Acidic Protein, Genetics, medicine, MESH: Recombinant Fusion Proteins, MESH: Plasmodium berghei, Animals, Humans, Molecular Biology, MESH: Mice, MESH : Plasmodium berghei, 030304 developmental biology, MESH: Humans, MESH : Cell Line, Tumor, MESH : Humans, MESH : Astrocytes, MESH : Astrocytoma, MESH: Immunohistochemistry, [SDV.IMM.IMM]Life Sciences [q-bio]/Immunology/Immunotherapy, Single-Domain Antibodies, central nervous system, Fusion protein, Molecular biology, MESH: Cell Line, MESH: Astrocytes, Mice, Inbred C57BL, MESH : Fluorescent Antibody Technique, MESH: Astrocytoma, MESH : Brain, Astrocytes, biology.protein, Mutagenesis, Site-Directed, Paratope, MESH : Animals, BBB, MESH: Female, 030217 neurology & neurosurgery
Abstract: International audience; Antibodies normally do not cross the blood-brain barrier (BBB) and cannot bind an intracellular cerebral antigen. We demonstrate here for the first time that a new class of antibodies can cross the BBB without treatment. Camelids produce native homodimeric heavy-chain antibodies, the paratope being composed of a single-variable domain called VHH. Here, we used recombinant VHH directed against human glial fibrillary acidic protein (GFAP), a specific marker of astrocytes. Only basic VHHs (e.g., pI=9.4) were able to cross the BBB in vitro (7.8 vs. 0% for VHH with pI=7.7). By intracarotid and intravenous injections into live mice, we showed that these basic VHHs are able to cross the BBB in vivo, diffuse into the brain tissue, penetrate into astrocytes, and specifically label GFAP. To analyze their ability to be used as a specific transporter, we then expressed a recombinant fusion protein VHH-green fluorescent protein (GFP). These "fluobodies" specifically labeled GFAP on murine brain sections, and a basic variant (pI=9.3) of the fusion protein VHH-GFP was able to cross the BBB and to label astrocytes in vivo. The potential of VHHs as diagnostic or therapeutic agents in the central nervous system now deserves attention.
Published: 2012

5. PPARγ contributes to PKM2 and HK2 expression in fatty liver

Author: Catherine Espeillac, Ganna Panasyuk, Akira Suzuki, Francisco Verdeguer, Lluis Fajas, Céline Chauvin, Jean-Sébastien Annicotte, Marc Foretz, Ludivine A. Pradelli, Jean-Yves Scoazec, Marco Pontoglio, Pascal Ferré, Mario Pende, Morris J. Birnbaum, Jean-Ehrland Ricci, Yasuo Horie, Centre de recherche Croissance et signalisation (UMR_S 845), 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 Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA), Institut de recherche en cancérologie de Montpellier (IRCM - U896 Inserm - UM1), CRLCC Val d'Aurelle - Paul Lamarque-Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 1 (UM1), Institut Cochin (IC UM3 (UMR 8104 / U1016)), 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 de Recherche des Cordeliers (CRC (UMR_S 872)), Université Paris Descartes - Paris 5 (UPD5)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre de Recherche en Cancérologie de Lyon (UNICANCER/CRCL), Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Anipath, UFR Médecine Lyon-RTH Laennec-UFR Médecine Lyon-RTH Laennec, Institute for Diabetes, Obesity and Metabolism, University of Pennsylvania, University of Pennsylvania [Philadelphia], Centre méditerranéen de médecine moléculaire (C3M), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Recherche en Cancérologie de Montpellier (IRCM - U1194 Inserm - UM), CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), Pathologies nutritionnelles et métaboliques : obésité et diabète, Université Pierre et Marie Curie - Paris 6 (UPMC)-IFR58-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), This work was supported by grants from the European Research Council, from Fondation de la Recherche Medicale (DEQ20061107956) and from Fondation Schlumberger pour l'Education et la Recherche to M.P., and from the Association pour la Recherche sur le Cancer to M.P. and J.-E.R. L.A.P received a fellowship from the Région Provence-Alpes-Cote-d'Azur and INSERM, C.E. from Ministere de Recherche et Technologies and from Fondation de la Recherche Medicale., We are grateful to the members of INSERM-U845 for support, and to David Sabatini, Stefano Fumagalli, Benoit Viollet, Fabienne Foufelle, Renaud Dentin, Pascal Pineau, Isabelle André-Schmutz, Olivier Danos and Anne Dejean for helpful discussions and sharing reagents. We thank Sophie Berissi, Dominique Chretien and Sylvie Fabrega for technical support., Centre de recherche Croissance et signalisation ( UMR_S 845 ), 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 Nice Sophia Antipolis ( UNS ), Université Côte d'Azur ( UCA ), Institut de recherche en cancérologie de Montpellier ( IRCM - U896 Inserm - UM1 ), Université Montpellier 1 ( UM1 ) -CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université de Montpellier ( UM ), Institut Cochin ( UM3 (UMR 8104 / U1016) ), Centre de Recherche des Cordeliers ( CRC (UMR_S 872) ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Centre de Recherche en Cancérologie de Lyon ( CRCL ), Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ) -Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ) -Anipath, 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)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Côte d'Azur (UCA), Akita University, Kyushu University, Université Montpellier 1 (UM1)-CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), Université Pierre et Marie Curie - Paris 6 (UPMC)-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 de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Tumeurs endocrines digestives : mécanismes de la tumorigenèse et de la progression tumorale, Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM), University of Pennsylvania, and FORETZ, Marc
Subjects: MESH : Fatty Liver, MESH : Immunohistochemistry, MESH : Insulin, General Physics and Astronomy, Peroxisome proliferator-activated receptor, MESH : Promoter Regions, Genetic, MESH: Thiazolidinediones, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, Mice, 0302 clinical medicine, Hexokinase, MESH : Cell Proliferation, Insulin, MESH: Hexokinase, MESH: Animals, MESH : Glycolysis, Promoter Regions, Genetic, MESH: Fatty Liver, [SDV.MHEP.EM] Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, chemistry.chemical_classification, 0303 health sciences, Multidisciplinary, Disease genetics, MESH: Gene Expression Regulation, Enzymologic, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, Immunohistochemistry, MESH : Mice, Transgenic, MESH : Thiazolidinediones, MESH : Proto-Oncogene Proteins c-akt, MESH : Thyroid Hormones, Biochemistry, 030220 oncology & carcinogenesis, MESH: Glycolysis, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Peroxisome proliferator-activated receptor delta, MESH: Membrane Proteins, Peroxisome proliferator-activated receptor alpha, MESH : Carrier Proteins, Clinical pharmacology, PPARGC1B, Glycolysis, MESH : PPAR gamma, Thyroid Hormones, Peroxisome proliferator-activated receptor gamma, MESH: Mice, Transgenic, Mice, Transgenic, MESH: Carrier Proteins, [SDV.CAN]Life Sciences [q-bio]/Cancer, MESH: Insulin, Biology, PKM2, Gene Expression Regulation, Enzymologic, Article, General Biochemistry, Genetics and Molecular Biology, MESH : Gene Expression Regulation, Enzymologic, 03 medical and health sciences, MESH: Thyroid Hormones, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], MESH: Cell Proliferation, MESH : Mice, MESH: Promoter Regions, Genetic, Animals, Humans, MESH: Mice, Liver diseases, Cell Proliferation, 030304 developmental biology, MESH: Humans, MESH: Proto-Oncogene Proteins c-akt, Liver receptor homolog-1, MESH : Humans, Membrane Proteins, MESH: Immunohistochemistry, General Chemistry, MESH : Hexokinase, Fatty Liver, PPAR gamma, MESH: PPAR gamma, chemistry, MESH : Membrane Proteins, Thiazolidinediones, MESH : Animals, Carrier Proteins, Proto-Oncogene Proteins c-akt, Pyruvate kinase
Abstract: Rapidly proliferating cells promote glycolysis in aerobic conditions, to increase growth rate. Expression of specific glycolytic enzymes, namely pyruvate kinase M2 and hexokinase 2, concurs to this metabolic adaptation, as their kinetics and intracellular localization favour biosynthetic processes required for cell proliferation. Intracellular factors regulating their selective expression remain largely unknown. Here we show that the peroxisome proliferator-activated receptor gamma transcription factor and nuclear hormone receptor contributes to selective pyruvate kinase M2 and hexokinase 2 gene expression in PTEN-null fatty liver. Peroxisome proliferator-activated receptor gamma expression, liver steatosis, shift to aerobic glycolysis and tumorigenesis are under the control of the Akt2 kinase in PTEN-null mouse livers. Peroxisome proliferator-activated receptor gamma binds to hexokinase 2 and pyruvate kinase M promoters to activate transcription. In vivo rescue of peroxisome proliferator-activated receptor gamma activity causes liver steatosis, hypertrophy and hyperplasia. Our data suggest that therapies with the insulin-sensitizing agents and peroxisome proliferator-activated receptor gamma agonists, thiazolidinediones, may have opposite outcomes depending on the nutritional or genetic origins of liver steatosis., Molecular factors, regulating the expression of specific glycolytic enzymes that favour biosynthetic processes, have remained unknown. Panasyuk et al. identify PPARγ as a novel transcription factor turning on pyruvate kinase M2 and hexokinase 2, which are frequently upregulated in pathophysiological growth.
Published: 2012

6. The Inflammatory Response in Acyl-CoA Oxidase 1 Deficiency (Pseudoneonatal Adrenoleukodystrophy)

Author: Pierre Andreoletti, Stéphane Mandard, Aurore Vluggens, Gérard Lizard, Sander Kersten, Janardan K. Reddy, H. I. El Hajj, Kévin Ragot, Ronald J.A. Wanders, H. R. Waterham, Mustapha Cherkaoui-Malki, Amsterdam Gastroenterology Endocrinology Metabolism, Laboratory Genetic Metabolic Diseases, Lipides - Nutrition - Cancer (U866) ( LNC ), Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon ( ENSBANA ), Laboratoire Bio-PeroxIL. Biochimie du Peroxysome, Inflammation et Métabolisme Lipidique ( Bio-PeroxIL ), Université de Bourgogne ( UB ), Departments of Clinical Chemistry and Pediatrics, University of Amsterdam [Amsterdam] ( UvA ) -Academic Medical Centre, Nutrition, Metabolism and Genomics Group, Wageningen University and Research Centre [Wageningen] ( WUR ), Academic Medical Center [Amsterdam] ( AMC ), University of Amsterdam [Amsterdam] ( UvA ) -University of Amsterdam [Amsterdam] ( UvA ) -Universiteit van Amsterdam ( UvA ), Institut National de la Santé et de la Recherche Médicale, the Conseil Régional de Bourgogne, the Ministère de l'Enseignement Supérieur et de la Recherche, and the Centre National de la Recherche Scientifique., Mandard, Stéphane, Lipides - Nutrition - Cancer (U866) (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), Laboratoire Bio-PeroxIL. Biochimie du Peroxysome, Inflammation et Métabolisme Lipidique (Bio-PeroxIL), Université de Bourgogne (UB), University of Amsterdam [Amsterdam] (UvA)-Academic Medical Centre, Wageningen University and Research [Wageningen] (WUR), Academic Medical Center - Academisch Medisch Centrum [Amsterdam] (AMC), and University of Amsterdam [Amsterdam] (UvA)-University of Amsterdam [Amsterdam] (UvA)-Universiteit van Amsterdam (UvA)
Subjects: MESH: Inflammation, peroxisomal disorders, MESH: Osteopontin, medicine.medical_treatment, MESH : Immunohistochemistry, MESH : Transcriptome, chemokine receptors, Voeding, Metabolisme en Genomica, 0302 clinical medicine, Endocrinology, MESH: Reverse Transcriptase Polymerase Chain Reaction, Acyl-CoA oxidase, multiple-sclerosis lesions, MESH : Osteopontin, MESH : Fatty Acids, Cells, Cultured, Oligonucleotide Array Sequence Analysis, [SDV.MHEP.EM] Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, 0303 health sciences, Oxidase test, MESH : Gene Expression Regulation, Reverse Transcriptase Polymerase Chain Reaction, Fatty Acids, MESH: Acyl-CoA Oxidase, MESH : Reverse Transcriptase Polymerase Chain Reaction, Peroxisome, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, [ SDV.MHEP.EM ] Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, Immunohistochemistry, MESH: Gene Expression Regulation, Metabolism and Genomics, 3. Good health, MESH: Fatty Acids, MESH : Oligonucleotide Array Sequence Analysis, Cytokine, Metabolisme en Genomica, ACOX1, Adrenoleukodystrophy, Nutrition, Metabolism and Genomics, MESH : Acyl-CoA Oxidase, medicine.symptom, Inflammation Mediators, MESH: Cells, Cultured, medicine.medical_specialty, MESH : Interleukin-8, MESH : Interleukin-6, MESH: Inflammation Mediators, Inflammation, Biology, in-vitro, MESH : Interleukin-1, MESH : Inflammation Mediators, 03 medical and health sciences, Voeding, Internal medicine, Peroxisomal disorder, nf-kappa-b, MESH : Cells, Cultured, MESH : Fibroblasts, medicine, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, gene, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Nutrition, 030304 developmental biology, VLAG, MESH: Humans, MESH : Inflammation, Interleukin-6, MESH: Transcriptome, Interleukin-8, MESH : Humans, MESH: Interleukin-1, MESH: Immunohistochemistry, Fibroblasts, medicine.disease, MESH: Interleukin-6, MESH: Interleukin-8, Gene Expression Regulation, MESH: Fibroblasts, MESH: Oligonucleotide Array Sequence Analysis, cells, Brief Reports, Osteopontin, microarray analysis, Acyl-CoA Oxidase, Transcriptome, interleukin-1, 030217 neurology & neurosurgery, x-linked adrenoleukodystrophy, Interleukin-1
Abstract: Among several peroxisomal neurodegenerative disorders, the pseudoneonatal adrenoleukodystrophy (P-NALD) is characterized by the acyl-coenzyme A oxidase 1 (ACOX1) deficiency, which leads to the accumulation of very-long-chain fatty acids ( VLCFA) and inflammatory demyelination. However, the components of this inflammatory process in P-NALD remain elusive. In this study, we used transcriptomic profiling and PCR array analyses to explore inflammatory gene expression in patient fibroblasts. Our results show the activation of IL-1 inflammatory pathway accompanied by the increased secretion of two IL-1 target genes, IL-6 and IL-8 cytokines. Human fibroblasts exposed to very-long-chain fatty acids exhibited increased mRNA expression of IL-1 alpha and IL-1 beta cytokines. Furthermore, expression of IL-6 and IL-8 cytokines in patient fibroblasts was down-regulated by MAPK, p38MAPK, and Jun N-terminal kinase inhibitors. Thus, the absence of acyl-coenzyme A oxidase 1 activity in P-NALD fibroblasts triggers an inflammatory process, in which the IL-1 pathway seems to be central. The use of specific kinase inhibitors may permit the modulation of the enhanced inflammatory status. (Endocrinology 153: 2568-2575, 2012)
Published: 2012

7. Coagulation factor X mediates adenovirus type 5 liver gene transfer in non-human primates (Microcebus murinus)

Author: Nadine Mestre-Francés, Angela C. Bradshaw, Daniel Henaff, Andrew H. Baker, Raul Alba, Jean-Michel Verdier, British Heart Foundation Glasgow Cardiovascular Research Centre (BHF GCRC), University of Glasgow-NHS Greater Glasgow and Clyde, Mécanismes moléculaires dans les démences neurodégénératives (MMDN), Université Montpellier 2 - Sciences et Techniques (UM2)-École Pratique des Hautes Études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Génétique Moléculaire de Montpellier (IGMM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), This work was supported by the European Commission FP7 BRAINCAV programme., European Project: 222992,EC:FP7:HEALTH,FP7-HEALTH-2007-B,BRAINCAV(2008), Université de Montpellier (UM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Institut National de la Santé et de la Recherche Médicale (INSERM)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL), Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM), British Heart Foundation Glasgow Cardiovascular Research Centre, University of Glasgow, Mécanismes moléculaires dans les démences neurodégénératives ( MMDN ), École pratique des hautes études ( EPHE ) -Université Montpellier 2 - Sciences et Techniques ( UM2 ) -Université de Montpellier ( UM ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut de Génétique Moléculaire de Montpellier ( IGMM ), Centre National de la Recherche Scientifique ( CNRS ) -Université de Montpellier ( UM ), European Project : 222992,EC:FP7:HEALTH,FP7-HEALTH-2007-B,BRAINCAV ( 2008 ), Université Montpellier 2 - Sciences et Techniques (UM2)-École pratique des hautes études (EPHE), Verdier, Jean-Michel, and NONHUMAN ADENOVIRUS VECTORS FOR GENE TRANSFER TO THE BRAIN - BRAINCAV - - EC:FP7:HEALTH2008-10-01 - 2013-03-31 - 222992 - VALID
Subjects: Liver cytology, MESH: Spleen, MESH : Immunohistochemistry, viruses, MESH: Factor X, MESH : Hepatocytes, MESH: Gene Targeting, medicine.disease_cause, MESH: Hepatocytes, chemistry.chemical_compound, Transduction (genetics), 0302 clinical medicine, MESH: Genetic Vectors, MESH: Animals, 0303 health sciences, biology, Factor X, Gene Transfer Techniques, Gene targeting, MESH : Protein Binding, MESH: Adenoviridae, Immunohistochemistry, MESH : Genetic Vectors, 3. Good health, MESH : Factor X, medicine.anatomical_structure, Liver, [ SDV.BBM.GTP ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], 030220 oncology & carcinogenesis, Hepatocyte, Gene Targeting, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Molecular Medicine, MESH: Cheirogaleidae, MESH: Genome, Viral, Cheirogaleidae, MESH : Gene Therapy, MESH : Genome, Viral, MESH : Heparan Sulfate Proteoglycans, Protein Binding, MESH : Spleen, Microcebus murinus, MESH : Cell Membrane, Genetic Vectors, Spleen, MESH: Gene Transfer Techniques, Genome, Viral, Article, Adenoviridae, 03 medical and health sciences, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Genetics, medicine, MESH: Protein Binding, Animals, MESH : Adenoviridae, MESH : Gene Transfer Techniques, Molecular Biology, 030304 developmental biology, MESH : Cheirogaleidae, Cell Membrane, MESH : Liver, MESH: Immunohistochemistry, Genetic Therapy, biochemical phenomena, metabolism, and nutrition, biology.organism_classification, Molecular biology, chemistry, MESH: Heparan Sulfate Proteoglycans, Hepatocytes, MESH : Animals, MESH: Gene Therapy, MESH : Gene Targeting, Heparan Sulfate Proteoglycans, MESH: Liver, MESH: Cell Membrane
Abstract: International audience; Coagulation factor X (FX)-binding ablated adenovirus type 5 (Ad5) vectors have been genetically engineered to ablate the interaction with FX, resulting in substantially reduced hepatocyte transduction following intravenous administration in rodents. Here, we quantify viral genomes and gene transfer mediated by Ad5 and FX-binding-ablated Ad5 vectors in non-human primates. Ad5 vectors accumulated in and mediated gene transfer predominantly to the liver, whereas FX-binding-ablated vectors primarily targeted the spleen but showed negligible liver gene transfer. In addition, we show that Ad5 binding to hepatocytes may be due to the presence of heparan sulfate proteoglycans (HSPGs) on the cell membrane. Therefore, the Ad5-FX-HSPG pathway mediating liver gene transfer in rodents is also the mechanism underlying Ad5 hepatocyte transduction in Microcebus murinus.
Published: 2011

8. Usher type 1G protein sans is a critical component of the tip-link complex, a structure controlling actin polymerization in stereocilia

Author: Dominique Weil, Sébastien Chardenoux, Paul Avan, Vincent Michel, Nicolas Michalski, Olinda Alegria-Prévot, Marcio Do Cruzeiro, Isabelle Perfettini, Guy P. Richardson, Richard J. Goodyear, Isabelle Foucher, Elisa Caberlotto, Jean-Pierre Hardelin, Amel Bahloul, Christine Petit, Elise Pepermans, Collège de France - Chaire Génétique et physiologie cellulaire, Collège de France (CdF (institution)), Université Pierre et Marie Curie - Paris 6 (UPMC), Génétique des Déficits Sensoriels, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), School of Life Sciences, University of Sussex, Plateforme Recombinaison Homologue, Transfert d'Embryons et Cryoconservation [Institut Cochin] (PRHTEC), Institut Cochin (IC UM3 (UMR 8104 / U1016)), 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), Equipe Biophysique Neurosensorielle [Neuro-Dol], Neuro-Dol (Neuro-Dol), Université d'Auvergne - Clermont-Ferrand I (UdA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université d'Auvergne - Clermont-Ferrand I (UdA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Neuro-Dol (Neuro-Dol), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Université Clermont Auvergne [2017-2020] (UCA [2017-2020]), This work was supported by FAUN Stiftung (Suchert Foundation), Fondation Raymonde and Guy Strittmatter,LHW-Stiftung, the Conny Maeva Charitable Foundation, Fondation Orange, and the Louis-Jeantet Foundation. R.J.G. and G.P.R. are supported by The Wellcome Trust, Chaire Génétique et physiologie cellulaire, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), michalski, nicolas, Génétique et Physiologie de l'Audition, Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Collège de France ( CdF ) -Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université Pierre et Marie Curie - Paris 6 ( UPMC ), Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Plateforme Recombinaison Homologue, Transfert d'Embryons et Cryoconservation [Institut Cochin] ( PRHTEC ), Institut Cochin ( UM3 (UMR 8104 / U1016) ), 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 ), Neuro-Dol ( Neuro-Dol ), Université d'Auvergne - Clermont-Ferrand I ( UdA ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université d'Auvergne - Clermont-Ferrand I ( UdA ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Neuro-Dol - Clermont Auvergne ( Neuro-Dol ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Clermont Auvergne ( UCA ) -Université Clermont Auvergne ( UCA ), and Collège de France ( CdF )
Subjects: Scaffold protein, MESH: Signal Transduction, MESH: Protein Precursors, MESH : Immunohistochemistry, [SDV]Life Sciences [q-bio], MESH : Actins, Fluorescent Antibody Technique, MESH : Analysis of Variance, MESH: Mice, Knockout, MESH: Cadherins, Polymerization, Mice, 0302 clinical medicine, MESH: Genetic Vectors, MESH: Hair Cells, Auditory, MESH : Polymerization, MESH: Animals, MESH: Nerve Tissue Proteins, Mechanotransduction, MESH : Protein Precursors, MESH : Nerve Tissue Proteins, MESH: Fluorescent Antibody Technique, Mice, Knockout, 0303 health sciences, Multidisciplinary, MESH: Electrophysiology, Cilium, Biological Sciences, Cadherins, Immunohistochemistry, MESH : Genetic Vectors, Cell biology, [SDV] Life Sciences [q-bio], Electrophysiology, medicine.anatomical_structure, MESH: Polymerization, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH : Cadherins, Transduction (physiology), Engineering sciences. Technology, Signal Transduction, MESH : Microscopy, Electron, Scanning, MESH: Microscopy, Electron, Scanning, Genetic Vectors, Cadherin Related Proteins, Nerve Tissue Proteins, Biology, MESH: Actins, MESH : Cilia, 03 medical and health sciences, MESH : Electrophysiology, MESH: Cilia, MESH: Analysis of Variance, MESH : Mice, Hair Cells, Auditory, medicine, otorhinolaryngologic diseases, Animals, [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Cilia, Protein Precursors, MESH : Hair Cells, Auditory, MESH: Mice, Actin, 030304 developmental biology, MESH : Signal Transduction, Analysis of Variance, [ SDV ] Life Sciences [q-bio], Stereocilia, MESH: Immunohistochemistry, Actins, MESH : Fluorescent Antibody Technique, Cytoplasm, [ SDV.NEU ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Microscopy, Electron, Scanning, MESH : Mice, Knockout, MESH : Animals, sense organs, Tip link, 030217 neurology & neurosurgery
Abstract: The mechanotransducer channels of auditory hair cells are gated by tip-links, oblique filaments that interconnect the stereocilia of the hair bundle. Tip-links stretch from the tips of stereocilia in the short and middle rows to the sides of neighboring, taller stereocilia. They are made of cadherin-23 and protocadherin-15, products of the Usher syndrome type 1 genes USH1D and USH1F, respectively. In this study we address the role of sans, a putative scaffold protein and product of the USH1G gene. In Ush1g −/− mice, the cohesion of stereocilia is disrupted, and both the amplitude and the sensitivity of the transduction currents are reduced. In Ush1g fl/fl Myo15-cre +/− mice, the loss of sans occurs postnatally and the stereocilia remain cohesive. In these mice, there is a decrease in the amplitude of the total transducer current with no loss in sensitivity, and the tips of the stereocilia in the short and middle rows lose their prolate shape, features that can be attributed to the loss of tip-links. Furthermore, stereocilia from these rows undergo a dramatic reduction in length, suggesting that the mechanotransduction machinery has a positive effect on F-actin polymerization. Sans interacts with the cytoplasmic domains of cadherin-23 and protocadherin-15 in vitro and is absent from the hair bundle in mice defective for either of the two cadherins. Because sans localizes mainly to the tips of short- and middle-row stereocilia in vivo, we conclude that it belongs to a molecular complex at the lower end of the tip-link and plays a critical role in the maintenance of this link.
Published: 2011

9. Human genetic polymorphisms in T1R1 and T1R3 taste receptor subunits affect their function

Author: Claire Schlegel-Le-Poupon, Annick Faurion, Jean-Claude Pernollet, Singh Preet Bano, Christine Belloir, Hélène Débat, Loïc Briand, Brice Hoffmann, Anna Wiencis, Maud Sigoillot, Claude Nespoulous, Marta Grauso, Jean-Pierre Montmayeur, Mariam Raliou, Didier Trotier, Briand, Loïc, Neurobiologie de l'Olfaction et de la Prise Alimentaire - Biochimie de l'Olfaction et de la Gustation, Institut National de la Recherche Agronomique (INRA), Centre des Sciences du Goût et de l'Alimentation [Dijon] (CSGA), Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique (CNRS), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Institut de Neurobiologie Alfred Fessard (INAF), Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique ( INRA ), Centre des Sciences du Goût et de l'Alimentation [Dijon] ( CSGA ), Institut National de la Recherche Agronomique ( INRA ) -Université de Bourgogne ( UB ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique ( CNRS ), Université Versailles St-Quentin, Institut de Neurobiologie Alfred Fessard ( INAF ), Centre National de la Recherche Scientifique ( CNRS ), and Neurobiologie de l'Olfaction et de la Prise Alimentaire (NOPA)
Subjects: Nonsynonymous substitution, Models, Molecular, Taste, 030309 nutrition & dietetics, Physiology, Monosodium glutamate, MESH : Polymorphism, Genetic, saccharin, MESH : Immunohistochemistry, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, récepteur gustatif, goût, Umami, MESH : Blotting, Western, MESH: Receptors, G-Protein-Coupled, perception, gustation, Receptors, G-Protein-Coupled, Behavioral Neuroscience, chemistry.chemical_compound, MESH : Taste Threshold, Taste receptor, taste receptor, Sodium Glutamate, structure cristalline, dynamique moléculaire, [ SDV.GEN.GH ] Life Sciences [q-bio]/Genetics/Human genetics, papillae, Cells, Cultured, Genetics, 0303 health sciences, molecular dynamic, MESH: Taste Threshold, mammifère, récepteur couplé aux protéines G, MESH: Amino Acid Substitution, umami, Immunohistochemistry, Sensory Systems, Transmembrane domain, Biochemistry, [ SDV.NEU.NB ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Alimentation et Nutrition, Taste Threshold, récepteur glutamate, MESH : Receptors, G-Protein-Coupled, MESH: Models, Molecular, MESH: Cells, Cultured, t1r3, crystal structure, t1r1, MESH : Sodium Glutamate, MESH : Models, Molecular, Blotting, Western, différence individuelle, G protein coupled receptor, glutamate, individual difference, mammalian sweet, 03 medical and health sciences, protein coupled receptor, MESH : Amino Acid Substitution, man, Physiology (medical), MESH : Cells, Cultured, MESH: Polymorphism, Genetic, saccharine, MESH: Blotting, Western, Food and Nutrition, Humans, mammals, l glutamate, human, metabotropic glutamate receptor, umami taste, 030304 developmental biology, G protein-coupled receptor, MESH: Humans, Polymorphism, Genetic, papilla, MESH: Sodium Glutamate, MESH : Humans, snp, MESH: Immunohistochemistry, gustatory receptor, molecular dynamics, chemistry, crystalline structure, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, Amino Acid Substitution, Metabotropic glutamate receptor, msg, saccharin preference, humain, papille
Abstract: International audience; Umami is the typical taste induced by monosodium glutamate (MSG), which is thought to be detected by the heterodimeric G protein-coupled receptor, T1R1 and T1R3. Previously, we showed that MSG detection thresholds differ substantially between individuals and we further showed that nontaster and hypotaster subjects are associated with nonsynonymous single polymorphisms occurring in the T1R1 and T1R3 genes. Here, we show using functional expression that both amino acid substitutions (A110V and R507Q) in the N-terminal ligand-binding domain of T1R1 and the 2 other ones (F749S and R757C), located in the transmembrane domain of T1R3, severely impair in vitro T1R1/T1R3 response to MSG. A molecular model of the ligand-binding region of T1R1/T1R3 provides a mechanistic explanation supporting functional expression data. The data presented here support causal relations between the genotype and previous in vivo psychophysical studies in human evaluating sensitivity to MSG.
Published: 2011

10. Antiviral activity of Bay 41-4109 on hepatitis B virus in humanized Alb-uPA/SCID mice

Author: Eric Giang, Jacques Belghiti, Serban Morosan, Sophie Berissi, Céline Lamant, Johan Neyts, Eva Wimmer, Gherard Puerstinger, Patrick Soussan, Lucie DaSilva, Olivier Hantz, Marie-Noëlle Brunelle, Laurent Hannoun, Dina Kremsdorf, Hélène Massinet, Nicolas Brezillon, IFR Laennec (IL), Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Université Jean Monnet [Saint-Étienne] (UJM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), neuroscience and barin technology, Service de chirurgie, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Beaujon [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Service de Chirurgie Digestive, Hépato-Bilio-pancréatique et Transplantation Hépatique [CHU Pitié-Salpétrière], CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Department of Microbiology and Immunology, Rega Institute for Medical Research, Catholic University of Leuven - Katholieke Universiteit Leuven (KU Leuven), equipe 16, Physiopathologie moléculaire et nouveaux traitements des hépatites virales, Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-IFR62-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-IFR62-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Recherche en Cancérologie de Lyon (UNICANCER/CRCL), Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Léon Bérard [Lyon]-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), Carcinogenèse Hépatique et Virologie Moléculaire, Institut National de la Santé et de la Recherche Médicale (INSERM), Pathogénèse des Hépatites Virales B et Immunothérapie (UMR_S 812), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), IFR Laennec ( IL ), Institut National de la Recherche Agronomique ( INRA ) -Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Université Jean Monnet [Saint-Étienne] ( UJM ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Beaujon, Service de chirurgie digestive et hépato-bilio-pancréatique [CHU Pitié-Salpétrière], Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Pitié-Salpêtrière [APHP], Catholic University of Leuven ( KU Leuven ), Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-IFR62-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-IFR62-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre de Recherche en Cancérologie de Lyon ( CRCL ), Centre Léon Bérard [Lyon]-Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ) -Centre Léon Bérard [Lyon]-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 ), Pathogénèse des Hépatites Virales B et Immunothérapie ( UMR_S 812 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Beaujon, Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Pitié-Salpêtrière [APHP], Université de Lyon-Université de Lyon-IFR62-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre de Recherche en Cancérologie de Lyon (CRCL), and Université de Lyon-Université de Lyon-Centre Léon Bérard [Lyon]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre Léon Bérard [Lyon]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Mouse, Pyridines, Gastroenterology and hepatology, Biopsy, medicine.medical_treatment, MESH : Immunohistochemistry, MESH : Pyrimidines, MESH : Hepatitis B virus, MESH : Viral Load, Mice, SCID, MESH : Hepatocytes, Liver transplantation, medicine.disease_cause, Hepatitis, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, MESH: Hepatocytes, Mice, MESH: Biopsy, 0302 clinical medicine, MESH : Albumins, MESH: Animals, MESH : DNA, Viral, MESH: Mice, SCID, 0303 health sciences, Multidisciplinary, MESH: Kinetics, virus diseases, Animal Models, Viral Load, Hepatitis B, Antivirals, Immunohistochemistry, 3. Good health, MESH : Antiviral Agents, Infectious hepatitis, MESH: Hepatitis B virus, Real-time polymerase chain reaction, Liver, Capsid, 030220 oncology & carcinogenesis, Infectious diseases, Medicine, MESH : Kinetics, MESH: Viral Load, Viral load, Research Article, MESH: Antiviral Agents, Hepatitis B virus, MESH : Pyridines, Science, Alpha interferon, Viremia, [SDV.CAN]Life Sciences [q-bio]/Cancer, Viral diseases, Biology, Antiviral Agents, Microbiology, 03 medical and health sciences, Model Organisms, Albumins, Virology, MESH : Mice, medicine, Animals, Humans, MESH: Mice, Liver diseases, 030304 developmental biology, MESH: Humans, MESH: Hepatitis B, MESH : Humans, MESH: Pyridines, MESH : Liver, MESH : Hepatitis B, MESH: Immunohistochemistry, medicine.disease, MESH : Mice, SCID, digestive system diseases, MESH: DNA, Viral, Kinetics, Animal Models of Infection, Pyrimidines, Viral replication, MESH : Biopsy, MESH: Pyrimidines, DNA, Viral, Immunology, Hepatocytes, MESH : Animals, MESH: Albumins, MESH: Liver
Abstract: International audience; Current treatments for HBV chronic carriers using interferon alpha or nucleoside analogues are not effective in all patients and may induce the emergence of HBV resistant strains. Bay 41-4109, a member of the heteroaryldihydropyrimidine family, inhibits HBV replication by destabilizing capsid assembly. The aim of this study was to determine the antiviral effect of Bay 41-4109 in a mouse model with humanized liver and the spread of active HBV. Antiviral assays of Bay 41-4109 on HepG2.2.15 cells constitutively expressing HBV, displayed an IC(50) of about 202 nM with no cell toxicity. Alb-uPA/SCID mice were transplanted with human hepatocytes and infected with HBV. Ten days post-infection, the mice were treated with Bay 41-4109 for five days. During the 30 days of follow-up, the HBV load was evaluated by quantitative PCR. At the end of treatment, decreased HBV viremia of about 1 log(10) copies/ml was observed. By contrast, increased HBV viremia of about 0.5 log(10) copies/ml was measured in the control group. Five days after the end of treatment, a rebound of HBV viremia occurred in the treated group. Furthermore, 15 days after treatment discontinuation, a similar expression of the viral capsid was evidenced in liver biopsies. Our findings demonstrate that Bay 41-4109 displayed antiviral properties against HBV in humanized Alb-uPA/SCID mice and confirm the usefulness of Alb-uPA/SCID mice for the evaluation of pharmaceutical compounds. The administration of Bay 41-4109 may constitute a new strategy for the treatment of patients in escape from standard antiviral therapy.
Published: 2011

11. Phenotyping the claudin 11 deficiency in testis: from histology to immunohistochemistry

Author: Brigitte Le Magueresse-Battistoni, Alexander Gow, Séverine Mazaud-Guittot, Groupe d'Etude de la Reproduction Chez l'Homme et les Mammiferes (GERHM), Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM), Cardiovasculaire, métabolisme, diabétologie et nutrition (CarMeN), Institut National de la Recherche Agronomique (INRA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Hospices Civils de Lyon (HCL)-Institut National de la Santé et de la Recherche Médicale (INSERM), NIDCD NIH HHS DC006262 R01 DC006262, Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-IFR140-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Hospices Civils de Lyon (HCL), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), Hospices Civils de Lyon (HCL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National des Sciences Appliquées de Lyon (INSA Lyon), Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université de Lyon-Institut National des Sciences Appliquées (INSA)-Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Institut National de la Recherche Agronomique (INRA), Groupe d'Etude de la Reproduction Chez l'Homme et les Mammiferes ( GERHM ), Université de Rennes 1 ( UR1 ), Université de Rennes ( UNIV-RENNES ) -Université de Rennes ( UNIV-RENNES ) -IFR140-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Cardiovasculaire, métabolisme, diabétologie et nutrition ( CarMeN ), Institut National de la Recherche Agronomique ( INRA ) -Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Institut National des Sciences Appliquées de Lyon ( INSA Lyon ), and Université de Lyon-Institut National des Sciences Appliquées ( INSA ) -Institut National des Sciences Appliquées ( INSA ) -Hospices Civils de Lyon ( HCL ) -Institut National de la Santé et de la Recherche Médicale ( INSERM )
Subjects: Male, Somatic cell, MESH: Spermatogonia, MESH : Immunohistochemistry, MESH : Eosine Yellowish-(YS), MESH: Formaldehyde, MESH: Mice, Knockout, MESH: Spermatocytes, MESH: Tight Junctions, Mice, MESH : Spermatogonia, 0302 clinical medicine, Spermatocytes, MESH : Fixatives, MESH : Microscopy, MESH: Animals, MESH: Fixatives, MESH: Nerve Tissue Proteins, Hematoxylin, MESH : Nerve Tissue Proteins, Blood–testis barrier, Mice, Knockout, 0303 health sciences, Microscopy, Claudin 11, 030219 obstetrics & reproductive medicine, Seminiferous Tubules, Sertoli cell, Immunohistochemistry, Cell biology, MESH : Spermatocytes, MESH : Phenotype, medicine.anatomical_structure, Phenotype, Seminiferous Epithelium, MESH : Seminiferous Tubules, MESH : Seminiferous Epithelium, Eosine Yellowish-(YS), Germ cell, MESH : Sertoli Cells, medicine.medical_specialty, Cell type, endocrine system, sertoli cell, MESH: Citric Acid, MESH: Microscopy, MESH : Infertility, MESH : Male, MESH: Seminiferous Tubules, Nerve Tissue Proteins, MESH : Citric Acid, blood testis barrier, Biology, MESH: Phenotype, Citric Acid, Article, Tight Junctions, 03 medical and health sciences, Fixatives, MESH: Sertoli Cells, Internal medicine, Formaldehyde, MESH : Mice, medicine, Animals, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Infertility, Claudin, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Mice, 030304 developmental biology, MESH : Hematoxylin, MESH : Tight Junctions, Sertoli Cells, MESH: Immunohistochemistry, Epithelium, Spermatogonia, spermatogenesis, MESH: Male, Endocrinology, Infertility, MESH : Formaldehyde, Claudins, MESH : Mice, Knockout, MESH: Eosine Yellowish-(YS), MESH: Hematoxylin, MESH : Animals, gene regulation, Spermatogenesis, MESH: Seminiferous Epithelium
Abstract: International audience; The testis is a heterogeneous organ that comprises a number of cell types, including germ cells at -different stages in their maturation, differentiated neighbor nursing cells, and endocrine somatic cells. Despite such cellular heterogeneity the testis is highly organized, with germ cell development and differentiation being compartmentalized into the interconnected tubular network of the seminiferous epithelium. Intratesticular scaffolds rely heavily on the basement membrane of the seminiferous tubules while germ cell development inside the seminiferous epithelium is critically dependent on the Blood Testis Barrier (BTB). The BTB is a macromolecular tight junction complex generated by somatic Sertoli cells within the seminiferous epithelium. The BTB divides the seminiferous epithelium into two compartments: the basal compartment, which delineates a niche for the proliferation and renewal of spermatogonia; and the adluminal compartment, where differentiating germ cells undergo meiosis and spermiogenesis. The BTB is unique in mammalian tissues because it is cyclically reconstructed during the spermatogenic cycle as preleptotene spermatocytes migrate from the basal compartment to the adluminal compartment and enter meiosis. In mouse, the loss of the BTB in the absence of the claudin 11 protein causes azoospermia and leads to infertility. Specifically, cldn11 deficiency results in sloughing of the cells of the seminiferous epithelium into the lumen. Understanding this pathophysiology has involved histological examination of the tissue defects as well as immunohistological characterization. Here, we present a comparative study of several modifications to the classical Hematoxylin-Eosin stain that may improve the diagnostic usefulness of this technique, as well as the use of several selective markers to identify testicular cell types.
Published: 2011

12. Molecular analysis and intestinal expression of SAR1 genes and proteins in Anderson's disease (Chylomicron retention disease)

Author: Marie Elisabeth Samson-Bouma, Lawrence P. Aggerbeck, Marianne Abifadel, Mathilde Varret, A. Morali, Jacqueline Champigneulle, Dominique Bonnefont-Rousselot, Jean P Rabès, Jean C Guenedet, Eric Bruckert, Thomas Aparicio, Amandine Georges, Catherine Boileau, J. Bonneau, Nutrition-Génétique et Exposition aux Risques Environnementaux (NGERE), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lorraine (UL), Service de Médecine Infantile III et Génétique Clinique [CHRU Nancy], Centre Hospitalier Régional Universitaire de Nancy (CHRU Nancy), Génétique et épigénétique des maladies métaboliques, neurosensorielles et du développement (Inserm U781), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de Biochimie Métabolique [CHU Pitié-Salpêtrière], CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Département de Biologie Expérimentale, Métabolique et Clinique, Université Paris Descartes - Paris 5 (UPD5), Laboratoire d'Anatomie et de Cytologie Pathologiques, Université Paris 13 (UP13), Service de biochimie, d'hormonologie et de génétique moléculaire [CHU Amrboise Paré], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Ambroise Paré [AP-HP], Service de Gastro-entérologie [Avicenne], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris 13 (UP13)-Hôpital Avicenne [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Service de Microscopie Electronique, Service d’Endocrinologie, Métabolisme et Prévention des Risques Cardio-Vasculaires [CHU Pitié-Salpêtrière], Pharmacologie, toxicologie et signalisation cellulaire (U747), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), These studies were supported by INSERM (AG, JB, JPR, MA, CB, AM, MV, MES), CNRS (LPA) and by grants from PHRC (AOMO6024), (JB, JPR, MA, CB, MV, MES) and ANR (ANR-08-GENO-002-01), (JB, JPR, MA, CB, MV, MES)., Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Université Paris 13 (UP13)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Avicenne [AP-HP], BMC, Ed., Service d'Endocrinologie, Métabolisme et Prévention des Maladies Cardio-vasculaires [CHU Pitié-Salpêtrière], Nutrition-Génétique et Exposition aux Risques Environnementaux ( NGERE ), Université de Lorraine ( UL ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Centre Hospitalier Régional Universitaire de Nancy ( CHRU Nancy ), Génétique et épigénétique des maladies métaboliques, neurosensorielles et du développement ( Inserm U781 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Pitié-Salpêtrière [APHP], Université Paris Descartes - Paris 5 ( UPD5 ), Université Paris 13 ( UP13 ), Service de Biochimie et Génétique Moléculaire, Université de Versailles Saint-Quentin-en-Yvelines ( UVSQ ) -Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Ambroise Paré, Université Paris 13 ( UP13 ) -Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Avicenne, Service d'endocrinologie-métabolisme [CHU Pitié-Salpêtrière], Physiologie Cellulaire des Regulations Hormonales, Nutritionnelles et Pharmacologiques, and 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 : Immunohistochemistry, lcsh:Medicine, [SDV.GEN] Life Sciences [q-bio]/Genetics, medicine.disease_cause, Hypobetalipoproteinemias, PCSK9 Gene, Exon, MESH : Exons, 0302 clinical medicine, MESH : Child, MESH: Child, MESH : Female, Genetics(clinical), Pharmacology (medical), MESH: Serine Endopeptidases, Intestinal Mucosa, Child, Genetics (clinical), MESH : Intestines, Medicine(all), 0303 health sciences, Mutation, biology, SAR1B Gene, Serine Endopeptidases, Exons, General Medicine, SAR1B, Immunohistochemistry, 3. Good health, MESH: Malabsorption Syndromes, MESH : Monomeric GTP-Binding Proteins, Female, Proprotein Convertases, MESH : Mutation, Proprotein Convertase 9, Chylomicron retention disease, MESH: Intestines, MESH: Mutation, Adolescent, MESH : Male, MESH : Hypobetalipoproteinemias, MESH: Monomeric GTP-Binding Proteins, 03 medical and health sciences, Malabsorption Syndromes, MESH : Serine Endopeptidases, MESH : Adolescent, medicine, Humans, MESH: Hypobetalipoproteinemias, Gene, Monomeric GTP-Binding Proteins, 030304 developmental biology, MESH: Adolescent, [SDV.GEN]Life Sciences [q-bio]/Genetics, MESH: Humans, Research, PCSK9, MESH : Humans, lcsh:R, MESH: Immunohistochemistry, medicine.disease, Molecular biology, MESH: Male, MESH : Malabsorption Syndromes, biology.protein, [ SDV.GEN ] Life Sciences [q-bio]/Genetics, MESH: Exons, MESH: Female, 030217 neurology & neurosurgery
Abstract: Background Anderson's disease (AD) or chylomicron retention disease (CMRD) is a very rare hereditary lipid malabsorption syndrome. In order to discover novel mutations in the SAR1B gene and to evaluate the expression, as compared to healthy subjects, of the Sar1 gene and protein paralogues in the intestine, we investigated three previously undescribed individuals with the disease. Methods The SAR1B, SAR1A and PCSK9 genes were sequenced. The expression of the SAR1B and SAR1A genes in intestinal biopsies of both normal individuals and patients was measured by RTqPCR. Immunohistochemistry using antibodies to recombinant Sar1 protein was used to evaluate the expression and localization of the Sar1 paralogues in the duodenal biopsies. Results Two patients had a novel SAR1B mutation (p.Asp48ThrfsX17). The third patient, who had a previously described SAR1B mutation (p.Leu28ArgfsX7), also had a p.Leu21dup variant of the PCSK9 gene. The expression of the SAR1B gene in duodenal biopsies from an AD/CMRD patient was significantly decreased whereas the expression of the SAR1A gene was significantly increased, as compared to healthy individuals. The Sar1 proteins were present in decreased amounts in enterocytes in duodenal biopsies from the patients as compared to those from healthy subjects. Conclusions Although the proteins encoded by the SAR1A and SAR1B genes are 90% identical, the increased expression of the SAR1A gene in AD/CMRD does not appear to compensate for the lack of the SAR1B protein. The PCSK9 variant, although reported to be associated with low levels of cholesterol, does not appear to exert any additional effect in this patient. The results provide further insight into the tissue-specific nature of AD/CMRD.
Published: 2011

13. 17alpha-ethinylestradiol disrupts the ontogeny of the forebrain GnRH system and the expression of brain aromatase during early development of zebrafish

Author: Yann Le Page, Mélanie Vosges, François Brion, Yves Combarnous, Jean-Marc Porcher, Bon Chu Chung, Olivier Kah, Institut National de l'Environnement Industriel et des Risques (INERIS), Interactions cellulaires et moléculaires (ICM), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS), Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), French Ministry of Ecology and Sustainable Development 189_09 DRC01, French National Research Agency, Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur] (IFCE)-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-IFR140-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES), Centre National de la Recherche Scientifique (CNRS)-Université de Tours-Institut Français du Cheval et de l'Equitation [Saumur]-Institut National de la Recherche Agronomique (INRA), Institut National de l'Environnement Industriel et des Risques ( INERIS ), Interactions cellulaires et moléculaires ( ICM ), Université de Rennes 1 ( UR1 ), Université de Rennes ( UNIV-RENNES ) -Université de Rennes ( UNIV-RENNES ) -IFR140-Centre National de la Recherche Scientifique ( CNRS ), Physiologie de la reproduction et des comportements [Nouzilly] ( PRC ), and Institut National de la Recherche Agronomique ( INRA ) -Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique ( CNRS )
Subjects: Time Factors, 17 alpha-Ethinylestradiol (EE2), MESH : Immunohistochemistry, Health, Toxicology and Mutagenesis, MESH: Neurons, Fluorescent Antibody Technique, MESH : Dose-Response Relationship, Drug, Endocrine Disruptors, 010501 environmental sciences, Endoplasmic Reticulum, Ethinyl Estradiol, 01 natural sciences, MESH: Dose-Response Relationship, Drug, Gonadotropin-Releasing Hormone, chemistry.chemical_compound, MESH: Prosencephalon, (Neuro-) endocrine disruption, MESH: Gonadotropin-Releasing Hormone, MESH : Aromatase, MESH: Animals, Aromatase, Zebrafish, MESH: Fluorescent Antibody Technique, MESH: Ethinyl Estradiol, Neurons, Regulation of gene expression, 0303 health sciences, MESH: Statistics, Nonparametric, MESH : Endoplasmic Reticulum, biology, MESH: Gene Expression Regulation, Enzymologic, Immunohistochemistry, Cell biology, MESH: Endocrine Disruptors, medicine.anatomical_structure, Xenoestrogen, Aromatase B (cyp19a1b), Gonadotropin, hormones, hormone substitutes, and hormone antagonists, MESH : Time Factors, medicine.medical_specialty, endocrine system, medicine.drug_class, Central nervous system, Danio, MESH : Ethinyl Estradiol, MESH : Zebrafish, MESH: Zebrafish Proteins, [ SDV.BBM.BM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Aquatic Science, Development, MESH : Neurons, Gene Expression Regulation, Enzymologic, Statistics, Nonparametric, MESH : Gene Expression Regulation, Enzymologic, 03 medical and health sciences, Prosencephalon, MESH : Zebrafish Proteins, MESH: Endoplasmic Reticulum, Internal medicine, medicine, Zebrafish (Danio rerio), Animals, MESH: Aromatase, MESH: Zebrafish, MESH : Statistics, Nonparametric, MESH : Prosencephalon, 030304 developmental biology, 0105 earth and related environmental sciences, Dose-Response Relationship, Drug, MESH: Time Factors, MESH: Immunohistochemistry, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Zebrafish Proteins, biology.organism_classification, MESH : Fluorescent Antibody Technique, Endocrinology, MESH : Endocrine Disruptors, chemistry, Forebrain, biology.protein, MESH : Animals, Gonadotropin-releasing-hormone (GnRH), MESH : Gonadotropin-Releasing Hormone
Abstract: International audience; Until now, studies dedicated to the actions of endocrine disrupting chemicals (EDCs) on the reproductive axis have been concerned with their effects at the gonadal level leaving their actions on neuroendocrine circuits controlling reproduction virtually unexplored. In vertebrates, gonadotropin-releasing hormone (GnRH) is the key factor controlling the activity of the reproductive axis. The development and functioning of GnRH neurons are finely tuned by a series of factors, notably sex steroids, making these neurons potential targets of EDCs, notably in aquatic species. By means of immunohistochemistry, we examined the effects of low levels of ethinylestradiol (EE2 0.02 nM, 0.1 nM, 0.5 nM), a potent synthetic estrogen, on early development (at 5, 10, 20, 30 days post-fertilization) of the forebrain GnRH neurons in a model fish species, the zebrafish (Danio rerio). In parallel, the ER-regulated expression of cytochrome P450 aromatase B (AroB) protein, which is encoded by the cyp19a1b gene, was precisely mapped at the brain and pituitary levels in developing control and EE2-exposed zebrafish. We show that EE2 disrupts the ontogeny of GnRH system by inducing an increase in the number of GnRH-ir neurons and GnRH fibers based on their immunoreactivity as well as a decrease in the size of the GnRH-ir soma and a modification of the migration profile of GnRH-ir neurons. Furthermore, we report a spectacular dose and time-dependent induction of AroB expression in radial glial cells of the developing brain further illustrating the extreme sensitivity of AroB to xenoestrogen and the relevance of AroB as biomarker of xenoestrogen effects on the central nervous system. Collectively, these original and relevant observations highlight the sensitivity of GnRH and AroB to a synthetic estrogen during embryogenesis. These data reinforce the need to further study the mechanisms underlying EDC effects on key neuroendocrine circuits involved in reproduction and brain development of vertebrates.
Published: 2010

14. Rescue of a dystrophin-like protein by exon skipping in vivo restores GABAA-receptor clustering in the hippocampus of the mdx mouse

Author: Olivier Danos, Carole Gruszczynski, Luis Garcia, Cyrille Vaillend, Carine Ros, Caroline Perronnet, Elise Peltekian, Aurélie Goyenvalle, Serge Laroche, Mécanismes cellulaires et moléculaires de la plasticité et de la mémoire, Centre de Neurosciences Paris-Sud (CNPS), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS)-Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11)-Centre National de la Recherche Scientifique (CNRS), Génétique et épigénétique des maladies métaboliques, neurosensorielles et du développement (Inserm U781), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Thérapie des maladies du muscle strié, Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Pierre et Marie Curie - Paris 6 (UPMC), Centre de Neurosciences Paris-Sud ( CNPS ), Université Paris-Sud - Paris 11 ( UP11 ) -Centre National de la Recherche Scientifique ( CNRS ) -Université Paris-Sud - Paris 11 ( UP11 ) -Centre National de la Recherche Scientifique ( CNRS ), Université Paris-Sud - Paris 11 ( UP11 ) -Centre National de la Recherche Scientifique ( CNRS ), Génétique et épigénétique des maladies métaboliques, neurosensorielles et du développement ( Inserm U781 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), and Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
Subjects: MESH : Dystrophin, Male, mdx mouse, MESH: Hippocampus, Duchenne muscular dystrophy, MESH : Immunohistochemistry, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH : Blotting, Western, MESH : Mice, Inbred mdx, Hippocampal formation, MESH : Hippocampus, Hippocampus, Polymerase Chain Reaction, Dystrophin, [ SDV.NEU.SC ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Cognitive Sciences, Mice, 0302 clinical medicine, MESH : Exons, Drug Discovery, MESH : Female, MESH: Animals, Muscular dystrophy, MESH : Polymerase Chain Reaction, MESH: Receptors, GABA-A, 0303 health sciences, [SDV.NEU.PC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Psychology and behavior, MESH: Mice, Inbred mdx, [SDV.NEU.SC]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Cognitive Sciences, Exons, Immunohistochemistry, Cell biology, [ SDV.NEU.NB ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, Molecular Medicine, Female, Original Article, musculoskeletal diseases, congenital, hereditary, and neonatal diseases and abnormalities, MESH : Male, Blotting, Western, MESH : Mice, Inbred C57BL, Biology, [ SDV.NEU.PC ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Psychology and behavior, 03 medical and health sciences, MESH: Dystrophin, MESH: Mice, Inbred C57BL, Utrophin, MESH : Mice, medicine, Genetics, MESH: Muscular Dystrophy, Duchenne, Animals, MESH: Blotting, Western, Molecular Biology, MESH: Mice, 030304 developmental biology, Pharmacology, MESH : Receptors, GABA-A, MESH: Polymerase Chain Reaction, MESH: Immunohistochemistry, medicine.disease, Receptors, GABA-A, Molecular biology, Exon skipping, MESH: Male, Mice, Inbred C57BL, Muscular Dystrophy, Duchenne, Synaptic plasticity, biology.protein, Mice, Inbred mdx, MESH : Animals, MESH: Exons, MESH : Muscular Dystrophy, Duchenne, MESH: Female, 030217 neurology & neurosurgery
Abstract: International audience; Dystrophin, the cytoskeletal protein whose defect is responsible for Duchenne muscular dystrophy (DMD), is normally expressed in both muscles and brain. Genetic loss of brain dystrophin in the mdx mouse model of DMD reduces the capacity for type A gamma-aminobutyric acid (GABA(A))-receptor clustering in central inhibitory synapses, which is thought to be a main molecular defect leading to brain and cognitive alterations in this syndrome. U7 small nuclear RNAs modified to encode antisense sequences and expressed from recombinant adeno-associated viral (rAAV) vectors have proven efficient after intramuscular injection to induce skipping of the mutated exon 23 and rescue expression of a functional dystrophin-like product in muscle tissues of mdx mice in vivo. Here, we report that intrahippocampal injection of a single dose of rAAV2/1-U7 can rescue substantial levels of brain dystrophin expression (15-25%) in mdx mice for months. This is sufficient to completely restore GABA(A)-receptor clustering in pyramidal and dendritic layers of CA1 hippocampus, suggesting exon-skipping strategies offer the prospect to investigate and correct both brain and muscle alterations in DMD. This provides new evidence that in the adult brain dystrophin is critical for the control of GABA(A)-receptor clustering, which may have an important role in activity-dependent synaptic plasticity in hippocampal circuits.
Published: 2010

15. Comprehensive diagnostic strategy for blood culture-negative endocarditis: a prospective study of 819 new cases

Author: Jean-Paul Casalta, Pierre-Edouard Fournier, Jean-Pierre Arzouni, Franck Thuny, Hervé Richet, Didier Raoult, Jean-Luc Mainardi, Marie Célard, Thierry Caus, Gilbert Habib, Frederic Collart, Max Maurin, Hubert Lepidi, Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes (URMITE), Institut de Recherche pour le Développement (IRD)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR48, INSB-INSB-Centre National de la Recherche Scientifique (CNRS), Service de cardiologie, Université de la Méditerranée - Aix-Marseille 2-Assistance Publique - Hôpitaux de Marseille (APHM)- Hôpital de la Timone [CHU - APHM] (TIMONE), Fédération de Microbiologie Clinique, Hôpital de la Timone [CHU - APHM] (TIMONE), Laboratoire Adaptation et pathogénie des micro-organismes [Grenoble] (LAPM), Centre National de la Recherche Scientifique (CNRS)-Université Joseph Fourier - Grenoble 1 (UJF), CHU Grenoble, Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble, Laboratoire de microbiologie, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Université Paris Descartes - Paris 5 (UPD5), Affectations Mineralisantes du Systeme Cardiovasculaire : Calcifications Arterielles et Valvulaires Aortiques, Université de Picardie Jules Verne (UPJV)-Institut National de la Santé et de la Recherche Médicale (INSERM), Chirurgie cardiaque, CHU Amiens-Picardie, Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes ( URMITE ), Institut de Recherche pour le Développement ( IRD ) -Aix Marseille Université ( AMU ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -IFR48, INSB-INSB-Centre National de la Recherche Scientifique ( CNRS ), Université de la Méditerranée - Aix-Marseille 2-Assistance Publique - Hôpitaux de Marseille ( APHM ) - Hôpital de la Timone [CHU - APHM] ( TIMONE ), Hôpital de la Timone [CHU - APHM] ( TIMONE ), Laboratoire Adaptation et pathogénie des micro-organismes [Grenoble] ( LAPM ), Université Joseph Fourier - Grenoble 1 ( UJF ) -Centre National de la Recherche Scientifique ( CNRS ), Université Joseph Fourier - Grenoble 1 ( UJF ) -CHU Grenoble, Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Européen Georges Pompidou [APHP] ( HEGP ) -Université Paris Descartes - Paris 5 ( UPD5 ), Université de Picardie Jules Verne ( UPJV ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université Joseph Fourier - Grenoble 1 (UJF)-Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Européen Georges Pompidou [APHP] (HEGP), Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Université Paris Descartes - Paris 5 (UPD5), and Institut des sciences biologiques (INSB-CNRS)-Institut des sciences biologiques (INSB-CNRS)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Male, MESH: Endocarditis, MESH : Molecular Sequence Data, MESH : Pathology, Molecular, MESH : Immunohistochemistry, MESH : Prospective Studies, MESH : Aged, MESH: Molecular Diagnostic Techniques, 030204 cardiovascular system & hematology, Polymerase Chain Reaction, Serology, 0302 clinical medicine, MESH: Aged, 80 and over, MESH : Child, Neoplasms, MESH: Autoimmune Diseases, MESH: Child, Blood culture, MESH : Female, MESH: Neoplasms, 030212 general & internal medicine, Prospective Studies, Pathology, Molecular, Child, MESH : Polymerase Chain Reaction, Aged, 80 and over, Immunoassay, MESH: Aged, MESH: Middle Aged, medicine.diagnostic_test, biology, Endocarditis, [SDV.BID.EVO]Life Sciences [q-bio]/Biodiversity/Populations and Evolution [q-bio.PE], Bacterial Infections, Middle Aged, MESH : Adult, MESH : Immunoassay, Immunohistochemistry, MESH : Endocarditis, 3. Good health, MESH : Bacterial Infections, Infectious Diseases, [ SDV.BID.EVO ] Life Sciences [q-bio]/Biodiversity/Populations and Evolution [q-bio.PE], Molecular Diagnostic Techniques, MESH: Young Adult, Female, MESH: Immunoassay, Bartonella Infection, Microbiology (medical), Bartonella, Adult, Adolescent, MESH: Bacterial Infections, MESH : Male, Molecular Sequence Data, MESH : Young Adult, Q fever, Autoimmune Diseases, Tropheryma whipplei, 03 medical and health sciences, Young Adult, MESH : Autoimmune Diseases, MESH : Adolescent, medicine, Humans, MESH : Middle Aged, MESH : Aged, 80 and over, Aged, MESH: Pathology, Molecular, MESH: Adolescent, MESH: Molecular Sequence Data, MESH: Humans, business.industry, MESH : Humans, MESH: Adult, MESH: Immunohistochemistry, MESH: Polymerase Chain Reaction, medicine.disease, biology.organism_classification, Coxiella burnetii, Virology, MESH : Neoplasms, MESH: Male, MESH: Prospective Studies, business, MESH: Female, MESH : Molecular Diagnostic Techniques
Abstract: International audience; BACKGROUND. Blood culture-negative endocarditis (BCNE) may account for up to 31% of all cases of endocarditis. METHODS. We used a prospective, multimodal strategy incorporating serological, molecular, and histopathological assays to investigate specimens from 819 patients suspected of having BCNE. RESULTS. Diagnosis of endocarditis was first ruled out for 60 patients. Among 759 patients with BCNE, a causative microorganism was identified in 62.7%, and a noninfective etiology in 2.5%. Blood was the most useful specimen, providing a diagnosis for 47.7% of patients by serological analysis (mainly Q fever and Bartonella infections). Broad-range polymerase chain reaction (PCR) of blood and Bartonella-specific Western blot methods diagnosed 7 additional cases. PCR of valvular biopsies identified 109 more etiologies, mostly streptococci, Tropheryma whipplei, Bartonella species, and fungi. Primer extension enrichment reaction and autoimmunohistochemistry identified a microorganism in 5 additional patients. No virus or Chlamydia species were detected. A noninfective cause of endocarditis, particularly neoplasic or autoimmune disease, was determined by histological analysis or by searching for antinuclear antibodies in 19 (2.5%) of the patients. Our diagnostic strategy proved useful and sensitive for BCNE workup. CONCLUSIONS. We highlight the major role of zoonotic agents and the underestimated role of noninfective diseases in BCNE. We propose serological analysis for Coxiella burnetii and Bartonella species, detection of antinuclear antibodies and rheumatoid factor as first-line tests, followed by specific PCR assays for T. whipplei, Bartonella species, and fungi in blood. Broad-spectrum 16S and 18S ribosomal RNA PCR may be performed on valvular biopsies, when available.
Published: 2010

16. ZO-1 is involved in trophoblastic cell differentiation in human placenta

Author: Danièle Evain-Brion, Graziello Geneau, Michael Grynberg, Jean-Louis Frendo, Diane Carette, Pascale Gerbaud, André Malassiné, Laurent Cronier, Sédami Gnidehou, Jean Guibourdenche, Guillaume Pidoux, La grossesse normale et pathologique: développement et fonctions du placenta et de l'utérus ( UMR_S 767 ), 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 ), PremUp Foundation, Institut de Recherche pour le Développement ( IRD ) -Université Paris-Sud - Paris 11 ( UP11 ) -Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Université Paris Diderot - Paris 7 ( UPD7 ) -CHI Créteil-Université Paris Descartes - Paris 5 ( UPD5 ) -Sorbonne Universités-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut de physiologie et biologie cellulaires ( IPBC ), Université de Poitiers-Centre National de la Recherche Scientifique ( CNRS ), Service de biochimie-hormonologie, Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Robert Debré-Université Paris Diderot - Paris 7 ( UPD7 ), Université Paris Descartes - Paris 5 ( UPD5 ), Caisse d'Assurance Maladie des Professions Libérales Province, Conseil Régional d'Ile de France, INSERM, La grossesse normale et pathologique: développement et fonctions du placenta et de l'utérus (UMR_S 767), 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), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Universités-Université Paris Descartes - Paris 5 (UPD5)-CHI Créteil-Institut de Recherche pour le Développement (IRD)-Université Paris-Sud - Paris 11 (UP11)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Diderot - Paris 7 (UPD7), Institut de physiologie et biologie cellulaires (IPBC), Université de Poitiers-Centre National de la Recherche Scientifique (CNRS), Université Paris Diderot - Paris 7 (UPD7)-Hôpital Robert Debré-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP), Université Paris Descartes - Paris 5 (UPD5), 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), Institut de Recherche pour le Développement (IRD)-Université Paris-Sud - Paris 11 (UP11)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Diderot - Paris 7 (UPD7)-CHI Créteil-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Robert Debré-Université Paris Diderot - Paris 7 (UPD7), 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 Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Université Paris Descartes - Paris 5 (UPD5)-CHI Créteil-Institut de Recherche pour le Développement (IRD)-Université Paris-Sud - Paris 11 (UP11)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Diderot - Paris 7 (UPD7)
Subjects: Small interfering RNA, Time Factors, Physiology, MESH: Trophoblasts, Cellular differentiation, MESH : Immunohistochemistry, Connexin, Chorionic Gonadotropin, MESH : Fluorescence Recovery After Photobleaching, 0302 clinical medicine, MESH: Pregnancy, MESH: Desmoplakins, Pregnancy, MESH: Gestational Age, MESH : Connexin 43, MESH : Female, Cells, Cultured, reproductive and urinary physiology, MESH : Chorionic Gonadotropin, 0303 health sciences, Cell fusion, Tight junction, MESH: Kinetics, Cell Differentiation, MESH : Protein Binding, Zona Occludens-1, MESH: Placenta, Immunohistochemistry, Cell biology, Trophoblasts, medicine.anatomical_structure, Intercellular Junctions, [ SDV.BDLR ] Life Sciences [q-bio]/Reproductive Biology, MESH : Placenta, 030220 oncology & carcinogenesis, embryonic structures, MESH : Intercellular Junctions, Female, RNA Interference, MESH: Membrane Proteins, MESH : Trophoblasts, MESH : Kinetics, MESH : Cell Differentiation, Fluorescence Recovery After Photobleaching, Protein Binding, MESH: Cells, Cultured, MESH : Time Factors, MESH: Cell Differentiation, trophoblast differentiation, placenta, MESH: RNA Interference, Gestational Age, Biology, MESH: Phosphoproteins, MESH : Immunoprecipitation, Adherens junction, MESH : Desmoplakins, 03 medical and health sciences, MESH: Chorionic Gonadotropin, MESH : Cells, Cultured, medicine, Humans, Immunoprecipitation, MESH: Protein Binding, MESH: Cell Shape, Cell Shape, 030304 developmental biology, MESH: Humans, cell fusion, MESH: Immunoprecipitation, MESH : Cell Fusion, MESH: Time Factors, MESH : Humans, Trophoblast, Membrane Proteins, MESH: Immunohistochemistry, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Cell Biology, Phosphoproteins, MESH: Connexin 43, Kinetics, MESH : Pregnancy, Membrane protein, Desmoplakins, MESH : Membrane Proteins, MESH: Cell Fusion, Connexin 43, Zonula Occludens-1 Protein, MESH : Phosphoproteins, MESH : Cell Shape, MESH : RNA Interference, MESH: Female, MESH: Fluorescence Recovery After Photobleaching, MESH: Intercellular Junctions, MESH : Gestational Age
Abstract: (IF : 4,23); International audience; Trophoblastic cell-cell fusion is an essential event required during human placental development. Several membrane proteins have been described to be directly involved in this process, including connexin 43 (Cx43), syncytin 1 (Herv-W env), and syncytin 2 (Herv-FRD env glycoprotein). Recently, zona occludens (ZO) proteins (peripheral membrane proteins associated with tight junctions, adherens junctions, and gap junctions) were shown to be involved in mouse placental development. Moreover, zona occludens 1 (ZO-1) was localized mainly at the intercellular boundaries between human trophoblastic cells. Therefore the role of ZO-1 in the dynamic process of human trophoblastic cell-cell fusion was investigated using primary trophoblastic cells in culture. In vitro as in situ, ZO-1 was localized mainly at the intercellular boundaries between trophoblastic cells where its expression substantially decreased during differentiation and during fusion. At the same time, Cx43 was localized at the interface of trophoblastic cells and its expression increased during differentiation. To determine a functional role for ZO-1 during trophoblast differentiation, small interfering RNA (siRNA) was used to knock down ZO-1 expression. Cytotrophoblasts treated with ZO-1 siRNA fused poorly, but interestingly, decreased Cx43 expression without altering the functionality of trophoblastic cell-cell communication as measured by relative permeability time constant determined using gap-FRAP experiments. Because kinetics of Cx43 and ZO-1 proteins show a mirror image, a potential association of these two proteins was investigated. By using coimmunoprecipitation experiments, a physical interaction between ZO-1 and Cx43 was demonstrated. These results demonstrate that a decrease in ZO-1 expression reduces human trophoblast cell-cell fusion and differentiation.
Published: 2010

17. Tenectin is a novel alphaPS2betaPS integrin ligand required for wing morphogenesis and male genital looping in Drosophila

Author: Anne-Laure Bougé, Thomas A. Bunch, Timmy Kendall, Stéphane Fraichard, Isabelle Chauvel, Hervé Bouhin, Centre des Sciences du Goût et de l'Alimentation [Dijon] (CSGA), Institut National de la Recherche Agronomique (INRA)-Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique (CNRS), Centre des Sciences du Goût et de l'Alimentation [Dijon] ( CSGA ), and Institut National de la Recherche Agronomique ( INRA ) -Université de Bourgogne ( UB ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique ( CNRS )
Subjects: Male, MESH: Extracellular Matrix Proteins, MESH: Drosophila, MESH : Immunohistochemistry, [ SDV.AEN ] Life Sciences [q-bio]/Food and Nutrition, Integrin, Ligands, Looping morphogenesis, Extracellular matrix, chemistry.chemical_compound, 0302 clinical medicine, MESH: Genitalia, Male, Morphogenesis, MESH: Ligands, Drosophila Proteins, Wings, Animal, MESH: Animals, Transgenes, In Situ Hybridization, 0303 health sciences, Extracellular Matrix Proteins, MESH : Genitalia, Male, MESH : Ligands, Integrin alpha Chains, Cell adhesion molecule, MESH : In Situ Hybridization, Immunohistochemistry, 3. Good health, Cell biology, Larva, MESH : Integrin alpha Chains, Adhesion, Drosophila, MESH : Mutation, MESH : Transgenes, Tenectin, Drosophila Protein, Ecdysone, MESH: Mutation, MESH: Drosophila Proteins, MESH : Male, MESH : Wing, MESH: Transgenes, Biology, Genitalia, Male, Article, 03 medical and health sciences, MESH : Extracellular Matrix Proteins, MESH: In Situ Hybridization, Animals, MESH : Drosophila, Cell adhesion, Molecular Biology, 030304 developmental biology, MESH : Larva, Metamorphosis, MESH: Integrin alpha Chains, Left–right asymmetry, MESH: Immunohistochemistry, Cell Biology, MESH : Drosophila Proteins, MESH: Wing, MESH: Male, MESH: Morphogenesis, chemistry, MESH : Morphogenesis, Mutation, biology.protein, MESH : Animals, MESH: Larva, [SDV.AEN]Life Sciences [q-bio]/Food and Nutrition, 030217 neurology & neurosurgery, Developmental Biology
Abstract: International audience; Morphogenesis of the adult structures of holometabolous insects is regulated by ecdysteroids and juvenile hormones and involves cell-cell interactions mediated in part by the cell surface integrin receptors and their extracellular matrix (ECM) ligands. These adhesion molecules and their regulation by hormones are not well characterized. We describe the gene structure of a newly described ECM molecule, tenectin, and demonstrate that it is a hormonally regulated ECM protein required for proper morphogenesis of the adult wing and male genitalia. Tenectin's function as a new ligand of the PS2 integrins is demonstrated by both genetic interactions in the fly and by cell spreading and cell adhesion assays in cultured cells. Its interaction with the PS2 integrins is dependent on RGD and RGD-like motifs. Tenectin's function in looping morphogenesis in the development of the male genitalia led to experiments that demonstrate a role for PS integrins in the execution of left-right asymmetry.
Published: 2010

18. In situ protein expression in tumour spheres: development of an immunostaining protocol for confocal microscopy

Author: Bruno Saubaméa, Dominique Bellet, Virginie Dangles-Marie, Louis-Bastien Weiswald, Sophie Richon, Jean-Marc Guinebretière, 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 ), Service de Pathologie, Hôpital René HUGUENIN (Saint-Cloud)-INSTITUT CURIE, Service de Médecine Nucléaire, INSTITUT CURIE-Hôpital René HUGUENIN (Saint-Cloud), 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 ), This work was supported by Ligue Contre le Cancer, Comité Ile-de-France (Grant RS 06/75-71, RS 07/75-45), Institut National du Cancer et Cancérôpole Ile de France ('COLOMETASTEM' and 'CSC cross platform network' projects), Philippe and Denis Bloch Cancer Research Grant, Patrick Roy Translational Medicine Grant, Sally Paget-Brown Translational Research Grant, Geneviève and Jean-Paul Driot Transformative Research grant. LBW was supported by a predoctoral fellowship from the Association d'Aide à la Recherche Cancérologique de Saint-Cloud and from The Association pour la Recherche sur le Cance, 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 Curie [Paris]-Hôpital René HUGUENIN (Saint-Cloud), 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), 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), BMC, Ed., 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), Hôpital René HUGUENIN (Saint-Cloud)-Institut Curie [Paris], 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), 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), 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 Diderot - Paris 7 (UPD7)
Subjects: Cancer Research, MESH : Immunohistochemistry, Cell, MESH: Flow Cytometry, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, law.invention, 0302 clinical medicine, law, Neoplasms, Image Processing, Computer-Assisted, MESH: Microscopy, Confocal, MESH: Neoplasms, AC133 Antigen, MESH: Antigens, CD, 0303 health sciences, Microscopy, Confocal, medicine.diagnostic_test, biology, MESH: Peptides, MESH : Peptides, MESH: Gene Expression Regulation, Neoplastic, Flow Cytometry, lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens, Immunohistochemistry, MESH: Image Processing, Computer-Assisted, 3. Good health, Cell biology, Gene Expression Regulation, Neoplastic, medicine.anatomical_structure, Technical Advance, Oncology, 030220 oncology & carcinogenesis, Neoplastic Stem Cells, MESH : Image Processing, Computer-Assisted, MESH: Cell Line, Tumor, MESH : Flow Cytometry, MESH : Gene Expression Regulation, Neoplastic, Confocal, MESH: Glycoproteins, [SDV.CAN]Life Sciences [q-bio]/Cancer, lcsh:RC254-282, Flow cytometry, MESH: Gene Expression Profiling, 03 medical and health sciences, MESH : Neoplastic Stem Cells, [SDV.CAN] Life Sciences [q-bio]/Cancer, Antigens, CD, Cancer stem cell, Confocal microscopy, Cell Line, Tumor, MESH : Antigens, CD, Genetics, medicine, Humans, MESH : Microscopy, Confocal, Glycoproteins, 030304 developmental biology, MESH: Humans, MESH : Cell Line, Tumor, Gene Expression Profiling, MESH : Gene Expression Profiling, MESH : Humans, CD44, MESH: Immunohistochemistry, MESH: Neoplastic Stem Cells, MESH : Neoplasms, MESH : Glycoproteins, Molecular biology, Cell culture, biology.protein, Peptides, Immunostaining
Abstract: Background Multicellular tumour sphere models have been shown to closely mimic phenotype characteristics of in vivo solid tumours, or to allow in vitro propagation of cancer stem cells (CSCs). CSCs are usually characterized by the expression of specific membrane markers using flow cytometry (FC) after enzymatic dissociation. Consequently, the spatial location of positive cells within spheres is not documented. Confocal microscopy is the best technique for the imaging of thick biological specimens after multi-labelling but suffers from poor antibody penetration. Thus, we describe here a new protocol for in situ confocal imaging of protein expression in intact spheroids. Methods Protein expression in whole spheroids (150 μm in diameter) from two human colon cancer cell lines, HT29 and CT320X6, has been investigated with confocal immunostaining, then compared with profiles obtained through paraffin immunohistochemistry (pIHC) and FC. Target antigens, relevant for colon cancer and with different expression patterns, have been studied. Results We first demonstrate that our procedure overcomes the well-known problem of antibody penetration in compact structures by performing immunostaining of EpCAM, a membrane protein expressed by all cells within our spheroids. EpCAM expression is detected in all cells, even the deepest ones. Likewise, antibody access is confirmed with CK20 and CD44 immunostaining. Confocal imaging shows that 100% of cells express β-catenin, mainly present in the plasma membrane with also cytoplasmic and nuclear staining, in agreement with FC and pIHC data. pIHC and confocal imaging show similar CA 19-9 cytoplasmic and membranar expression profile in a cell subpopulation. CA 19-9+ cell count confirms confocal imaging as a highly sensitive method (75%, 62% and 51%, for FC, confocal imaging and pIHC, respectively). Finally, confocal imaging reveals that the weak expression of CD133, a putative colon CSC marker, is restricted to the luminal cell surface of colorectal cancer acini, with CD133+ cellular debris into glandular lumina. Conclusion The present protocol enables in situ visualization of protein expression in compact three-dimensional models by whole mount confocal imaging, allowing the accurate localization and quantification of cells expressing specific markers. It should prove useful to study rare events like CSCs within tumour spheres.
Published: 2010

19. A new rat model of prenatal bowel obstruction: development and early assessment

Author: Frédérique Sauvat, Franck Sturtz, Laurent Fourcade, Nadine Cerf-Bensussan, Sabine Sarnacki, Yoanne Mousseau, Naziha Khen-Dunlop, Biomolécules Thérapies anti-tumorales (EA4021), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503), Service de chirurgie pédiatrique viscérale, orthopédique et plastique [CHU Limoges], CHU Limoges, CHU Necker - Enfants Malades [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Interactions de l'épithelium intestinal avec le système immunitaire, Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de Biochimie et Génétique Moléculaire [CHU Limoges], Biomolécules Thérapies anti-tumorales ( EA4021 ), Université de Limoges ( UNILIM ) -Génomique, Environnement, Immunité, Santé, Thérapeutique ( GEIST FR CNRS 3503 ), and Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM )
Subjects: Pathology, MESH : Immunohistochemistry, medicine.medical_treatment, MESH: Random Allocation, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, [ SDV.MHEP.PED ] Life Sciences [q-bio]/Human health and pathology/Pediatrics, MESH : Intestinal Obstruction, Gastroenterology, Random Allocation, 0302 clinical medicine, MESH: Pregnancy, Pregnancy, Reference Values, Prenatal Diagnosis, Medicine, MESH : Female, MESH: Animals, [ SDV.MHEP.CHI ] Life Sciences [q-bio]/Human health and pathology/Surgery, [ SDV.GEN.GH ] Life Sciences [q-bio]/Genetics/Human genetics, MESH : Rats, Mortality rate, Intestinal atresia, MESH: Reference Values, General Medicine, MESH : Survival Rate, Immunohistochemistry, 3. Good health, Bowel obstruction, Survival Rate, Fetal Diseases, MESH: Fetal Diseases, [ SDV.BBM.GTP ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], [ SDV.NEU.NB ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, 030220 oncology & carcinogenesis, Gestation, 030211 gastroenterology & hepatology, Female, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH : Sensitivity and Specificity, medicine.medical_specialty, MESH: Rats, MESH: Survival Rate, MESH: Probability, MESH : Probability, Rat model, [ SDV.BBM.BM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, [SDV.MHEP.CHI]Life Sciences [q-bio]/Human health and pathology/Surgery, MESH : Rats, Wistar, MESH : Reference Values, Sensitivity and Specificity, MESH : Random Allocation, 03 medical and health sciences, Internal medicine, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Animals, Rats, Wistar, MESH: Prenatal Diagnosis, MESH : Prenatal Diagnosis, Probability, Fetus, [SDV.MHEP.PED]Life Sciences [q-bio]/Human health and pathology/Pediatrics, business.industry, Fetal surgery, MESH : Fetal Diseases, MESH: Immunohistochemistry, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, MESH: Rats, Wistar, medicine.disease, MESH : Disease Models, Animal, MESH: Sensitivity and Specificity, Rats, MESH : Pregnancy, Disease Models, Animal, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, [ SDV.NEU ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Atresia, Pediatrics, Perinatology and Child Health, Surgery, MESH : Animals, MESH: Disease Models, Animal, MESH: Intestinal Obstruction, business, MESH: Female, Intestinal Obstruction
Abstract: International audience; PURPOSE: Although intestinal motility disorders often complicate the postoperative surgical management of newborns with congenital intestinal atresia, their pathogenesis remains unclear. Animal models of prenatal intestinal obstruction have been mainly developed in the lamb and the chicken. Despite new insights brought by these models, they have one or more limitations, such as high fetal mortality rates, high costs, long gestation periods, and an insufficient number of fetuses per litter. Moreover, some species are phylogenetically distant from mammals. METHODS: We developed a reproducible model of prenatal intestinal obstruction in the rat to study the histologic changes induced by the obstruction. We report, the technical devices and the first assessment of this atresia model in a didactic way to allow other researchers to easily reproduce the model. RESULTS: Prenatal intestinal obstructions in this study fulfilled all the macroscopic and histologic criteria usually listed by other models of prenatal intestinal obstruction that have been developed in other species. Furthermore with our model, we obtained a high success rate at a low cost. CONCLUSIONS: We presented in this study a reproducible model of prenatal intestinal obstruction in the rat with the macroscopical and histologic features of prenatal intestinal obstruction.
Published: 2010

20. Brain growth receptors control lifespan

Author: Annick Blaise, Jacques Epelbaum, Laurent Kappeler, Pascale Cervera, Carlos De Magalhaes Filho, Catherine Loudes, Yves Le Bouc, Patricia Leneuve, Joëlle Dupont, Martin Holzenberger, Rüdiger Klein, Laurence Perin, Centre de Recherche Saint-Antoine (UMRS893), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Physiologie de la reproduction et des comportements [Nouzilly] (PRC), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur] (IFCE)-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS), Service d'anatomie et cytologie pathologiques [CHU Saint-Antoine], Université Pierre et Marie Curie - Paris 6 (UPMC)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), Neurobiologie de la Croissance et de la Senescence, Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Department of Molecular Neurobiology [Martinsried], Max Planck Institute of Neurobiology (MPIN), Max-Planck-Gesellschaft-Max-Planck-Gesellschaft, Funding Agence National pour la Recherche (grant NT05-3 42491) and European Union Network of Excellence LifeSpan (036894), Le Ministère de l'Education Nationale, de la Recherche et de la Technologie (MENRT), and Groupement d'Intérêt Scientifique Institut du Vieillissement sponsored this study with grants to MH and YLB. ARC, Inserm, and MENRT supported LK and CDMF., Autard, Delphine, Centre de Recherche Saint-Antoine ( UMRS893 ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Physiologie de la reproduction et des comportements [Nouzilly] ( PRC ), Institut National de la Recherche Agronomique ( INRA ) -Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique ( CNRS ), Service d'anatomie et cytologie pathologiques, Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Saint-Antoine [APHP], Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Max Planck Institute of Neurobiology ( MPIN ), Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université de Tours-Institut Français du Cheval et de l'Equitation [Saumur]-Institut National de la Recherche Agronomique (INRA), Université Pierre et Marie Curie - Paris 6 (UPMC)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Saint-Antoine [APHP], CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), and Institut National de la Recherche Agronomique (INRA)-Institut Français du Cheval et de l'Equitation [Saumur]-Université de Tours (UT)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Nervous system, Male, Pituitary gland, medicine.medical_treatment, MESH : Insulin-Like Growth Factor I, MESH : Blotting, Western, Somatoliberin, Mice, 0302 clinical medicine, MESH: Receptor, IGF Type 1, MESH: Animals, Biology (General), 0303 health sciences, MESH : Animals, Newborn, MESH : Receptor, IGF Type 1, MESH : Reverse Transcriptase Polymerase Chain Reaction, Brain, mammifère, mus musculus, Growth hormone–releasing hormone, Immunohistochemistry, Diabetes and Endocrinology, Somatostatin, Hypothalamus, neuroendocrinologie, General Agricultural and Biological Sciences, medicine.medical_specialty, QH301-705.5, Central nervous system, Blotting, Western, Longevity, MESH : Growth Hormone, hormone de croissance, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Endocrine system, MESH: Blotting, Western, [SDV.NEU] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], IGF, système nerveux, MESH : Body Temperature, MESH: Fertility, Neurosecretory Systems, Mice, Inbred C57BL, MESH : Energy Metabolism, Endocrinology, MESH : Brain, MESH: Growth Hormone, MESH: Female, 030217 neurology & neurosurgery, Hormone, Developmental Biology, MESH: Body Temperature, Physiology, MESH : Immunohistochemistry, MESH: Insulin-Like Growth Factor I, MESH : Longevity, souris, MESH: Mice, Knockout, MESH: Animals, Newborn, Body Temperature, Receptor, IGF Type 1, MESH : Fertility, MESH: Reverse Transcriptase Polymerase Chain Reaction, MESH : Female, Insulin-Like Growth Factor I, Receptor, Mice, Knockout, Reverse Transcriptase Polymerase Chain Reaction, General Neuroscience, MESH: Energy Metabolism, medicine.anatomical_structure, MESH: Longevity, Synopsis, Female, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Research Article, Somatotropic cell, MESH : Male, MESH : Mice, Inbred C57BL, Biology, MESH: Brain, MESH: Mice, Inbred C57BL, Internal medicine, MESH : Mice, medicine, Animals, croissance animale, MESH: Mice, 030304 developmental biology, General Immunology and Microbiology, Growth factor, MESH: Immunohistochemistry, Genetics and Genomics, MESH: Male, Fertility, Animals, Newborn, MESH: Neurosecretory Systems, Growth Hormone, [ SDV.NEU ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH : Mice, Knockout, MESH : Animals, Energy Metabolism, MESH : Neurosecretory Systems
Abstract: Mutations that decrease insulin-like growth factor (IGF) and growth hormone signaling limit body size and prolong lifespan in mice. In vertebrates, these somatotropic hormones are controlled by the neuroendocrine brain. Hormone-like regulations discovered in nematodes and flies suggest that IGF signals in the nervous system can determine lifespan, but it is unknown whether this applies to higher organisms. Using conditional mutagenesis in the mouse, we show that brain IGF receptors (IGF-1R) efficiently regulate somatotropic development. Partial inactivation of IGF-1R in the embryonic brain selectively inhibited GH and IGF-I pathways after birth. This caused growth retardation, smaller adult size, and metabolic alterations, and led to delayed mortality and longer mean lifespan. Thus, early changes in neuroendocrine development can durably modify the life trajectory in mammals. The underlying mechanism appears to be an adaptive plasticity of somatotropic functions allowing individuals to decelerate growth and preserve resources, and thereby improve fitness in challenging environments. Our results also suggest that tonic somatotropic signaling entails the risk of shortened lifespan., Author Summary Using a mouse model relevant for humans, we showed that lifespan can be significantly extended by reducing the signaling selectively of a protein called IGF-I in the central nervous system. This effect occurred through changes in specific neuroendocrine pathways. Dissecting the pathophysiological mechanism, we discovered that IGF receptors in the mammalian brain efficiently steered the development of the somatotropic axis, which in turn affected the individual growth trajectory and lifespan. Our work confirms experimentally that continuously low IGF-I and low growth hormone levels favor extended lifespan and postpone age-related mortality. Together with other recent reports, our results further challenge the view that administration of GH can prevent, or even counteract human aging. This knowledge is important since growth hormone is often prescribed to elderly people in an attempt to compensate the unwanted effects of aging. Growth hormone and IGF-I are also substances frequently used for doping in sports., Inactivating IGF receptors in the brain decreased growth hormone and IGF-I, and increased lifespan in healthy mice. Such neuroendocrine longevity could be a physiological response to environment.
Published: 2010

21. Chronic infection during placental malaria is associated with up-regulation of cycloxygenase-2

Author: Emilie Fréalle, Odile Mercereau-Puijalon, Hamoud O Brahim, Alioune Gaye, Laurence Marrama, Jean Marie Dangou, Jean Y. Lehesran, Demba Sarr, Makhtar Niang, Ronan Jambou, Delphine Aldebert, Institut Pasteur de Dakar, Réseau International des Instituts Pasteur (RIIP), Centre de Santé Roi Baudoin, Immunologie moléculaire des parasites, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), UR10, Mother & Child Unit, Mother & Child Unit, Santé de la mère et de l'enfant en milieu tropical : épidémiologie génétique et périnatale, Université Paris Descartes - Paris 5 (UPD5), Département de Parasitologie Mycologie, Institut Pasteur [Paris] (IP), Institut Pasteur de Dakar - Réseau International des Instituts Pasteur (RIIP), Institut Pasteur [Paris] - Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris], and Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)
Subjects: MESH: Inflammation, MESH : RNA, Messenger, Placenta Diseases, Placenta, 0302 clinical medicine, MESH: Pregnancy, Pregnancy, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH : Pregnancy Complications, Parasitic, MESH: Up-Regulation, MESH : Up-Regulation, 0303 health sciences, MESH : Reverse Transcriptase Polymerase Chain Reaction, Pregnancy Outcome, MESH: Placenta, MESH : Senegal, Immunohistochemistry, MESH: Case-Control Studies, 3. Good health, Up-Regulation, Interleukin 10, MESH : Placenta, MESH: Young Adult, MESH : Infant, Low Birth Weight, MESH : Case-Control Studies, lcsh:RC955-962, MESH : Cyclooxygenase 2, MESH : Young Adult, MESH: Infant, Low Birth Weight, lcsh:Infectious and parasitic diseases, 03 medical and health sciences, MESH: Senegal, MESH : Adolescent, Humans, RNA, Messenger, MESH: Adolescent, MESH : Inflammation, MESH: Humans, MESH : Humans, MESH: Adult, medicine.disease, MESH : Pregnancy, MESH : Biopsy, Case-Control Studies, Immunology, Parasitology, Cyclooxygenase, MESH: Female, Malaria, MESH: Interleukin-10, MESH : Immunohistochemistry, Biopsy, MESH : Malaria, Falciparum, MESH: Pregnancy Complications, Parasitic, MESH: Placenta Diseases, MESH: Biopsy, MESH: Reverse Transcriptase Polymerase Chain Reaction, MESH : Female, Malaria, Falciparum, MESH : Pregnancy Outcome, biology, Reverse Transcriptase Polymerase Chain Reaction, MESH: Malaria, Falciparum, MESH: Infant, Newborn, MESH : Adult, Senegal, Interleukin-10, Infectious Diseases, medicine.anatomical_structure, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, Female, medicine.symptom, MESH: Cyclooxygenase 2, Adult, lcsh:Arctic medicine. Tropical medicine, Adolescent, 030231 tropical medicine, MESH : Placenta Diseases, Inflammation, MESH : Infant, Newborn, Young Adult, MESH : Interleukin-10, medicine, lcsh:RC109-216, 030304 developmental biology, MESH: RNA, Messenger, Research, Infant, Newborn, MESH: Immunohistochemistry, Infant, Low Birth Weight, MESH: Pregnancy Outcome, Chronic infection, Cyclooxygenase 2, Pregnancy Complications, Parasitic, biology.protein
Abstract: BackgroundPlacental malaria (PM) is associated with poor foetal development, but the pathophysiological processes involved are poorly understood. Cyclooxygenase (COX) and lipoxygenase (LOX) which convert fatty acids to prostaglandins and leukotrienes, play important roles in pregnancy and foetal development. COX-2, currently targeted by specific drugs, plays a dual role as it associates with both pre-eclampsia pathology and recovery during infection. The role of COX during PM was questioned by quantifying at delivery COX-1, COX-2, 15-LOX, and IL-10 expression in two groups of malaria infected and uninfected placenta.MethodsPlacental biopsies were collected at delivery for mRNA isolation and quantification, using real time PCR.ResultsCOX-2 and IL-10 mRNAs increased mainly during chronic infections (nine- and five-times, respectively), whereas COX-1 transcripts remained constant. COX-2 over-expression was associated with a higher birth weight of the baby, but with a lower rate of haemoglobin of the mother. It was associated with a macrophage infiltration of the placenta and with a low haemozoin infiltration. In the opposite way, placental infection was associated with lower expression of 15-LOX mRNA. A high degree of haemozoin deposition correlates with low birth weight and decreased expression of COX-2.ConclusionThese data provide evidence that COX-2 and IL-10 are highly induced during chronic infection of the placenta, but were not associated with preterm delivery or low birth weight. The data support the involvement of COX-2 in the recovery phase of the placental infection.
Published: 2010

22. Double immunohistochemistry with horseradish peroxidase and alkaline phosphatase detection systems

Author: Vincent Sarrazy, Desmoulière, A., Université de Limoges ( UNILIM ), Homéostasie Cellulaire et Pathologies ( HCP ), Université de Limoges ( UNILIM ) -CHU Limoges-Génomique, Environnement, Immunité, Santé, Thérapeutique ( GEIST FR CNRS 3503 ), Université de Limoges (UNILIM), Homéostasie Cellulaire et Pathologies (HCP), and Université de Limoges (UNILIM)-CHU Limoges-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503)
Subjects: [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH : Immunohistochemistry, MESH : Alkaline Phosphatase, [ SDV.BBM.BM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, In Vitro Techniques, MESH: Alkaline Phosphatase, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Animals, Humans, [ SDV.IMM ] Life Sciences [q-bio]/Immunology, MESH: Animals, [ SDV.GEN.GH ] Life Sciences [q-bio]/Genetics/Human genetics, MESH: Horseradish Peroxidase, Horseradish Peroxidase, MESH: Humans, MESH : Humans, MESH: Immunohistochemistry, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, MESH : Horseradish Peroxidase, Alkaline Phosphatase, Immunohistochemistry, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, [ SDV.BBM.GTP ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], [ SDV.NEU.NB ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, [ SDV.NEU ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], [SDV.IMM]Life Sciences [q-bio]/Immunology, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH : Animals
Abstract: Books Title : Histology Protocols; International audience; We describe here a protocol optimized for formalin-fixed paraffin-embedded tissue sections that enables the detection of two antigens. This technique allows immunohistochemistry to be performed with detection systems allowing observation by light microscopy. This chapter discusses the choice of appropriate protocols as well as the choice of visualization systems.In doing so, we provide examples of representative results obtained with this protocol and describe necessary controls; additionally, we discuss common problems associated with this methodology, and detail troubleshooting recommendations. Although this method has been optimized for liver sections, it may be applicable for performing double immunostaining in a variety of tissue samples.
Published: 2010

23. Evaluation of the collaborative network of highly correlating skin proteins and its change following treatment with glucocorticoids

Author: Nicolette Farman, Anette Fiebeler, Uwe Klinge, BMC, Ed., Surgical Department, Rheinisch-Westfälische Technische Hochschule Aachen University (RWTH), Department for applied medical engineering, Centre de Recherche des Cordeliers (CRC (UMR_S 872)), Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Clinics for Nephrology and Hypertension, Hannover Medical School [Hannover] (MHH), Rheinisch-Westfälische Technische Hochschule Aachen (RWTH), RWTH Aachen University Hospital, Germany, Centre de Recherche des Cordeliers ( CRC (UMR_S 872) ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Hannover Medical School [Hannover] ( MHH ), and Université Paris Descartes - Paris 5 (UPD5)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)
Subjects: MESH : Immunohistochemistry, Collaborative network, Bioinformatics, MESH : Proteins, Extracellular matrix, Mice, 0302 clinical medicine, MESH : Female, MESH: Animals, MESH: Proteins, [ SDV.BIBS ] Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], lcsh:QH301-705.5, Skin, [INFO.INFO-BI] Computer Science [cs]/Bioinformatics [q-bio.QM], 0303 health sciences, [SDV.BIBS] Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], Immunohistochemistry, [SDV.BIBS]Life Sciences [q-bio]/Quantitative Methods [q-bio.QM], 3. Good health, 030220 oncology & carcinogenesis, Modeling and Simulation, lcsh:R858-859.7, Female, Systems biology, Health Informatics, Biology, lcsh:Computer applications to medicine. Medical informatics, MESH: Multivariate Analysis, 03 medical and health sciences, MESH: Skin, Modelling and Simulation, [ INFO.INFO-BI ] Computer Science [cs]/Bioinformatics [q-bio.QM], MESH : Glucocorticoids, MESH : Mice, MESH : Skin, Animals, Mode of action, Glucocorticoids, MESH: Mice, 030304 developmental biology, Research, Proteins, MESH : Multivariate Analysis, MESH: Immunohistochemistry, Protein expression profile, lcsh:Biology (General), Treatment modality, Multivariate Analysis, MESH : Animals, [INFO.INFO-BI]Computer Science [cs]/Bioinformatics [q-bio.QM], MESH: Glucocorticoids, MESH: Female
Abstract: Background Glucocorticoids (GC) represent the core treatment modality for many inflammatory diseases. Its mode of action is difficult to grasp, not least because it includes direct modulation of many components of the extracellular matrix as well as complex anti-inflammatory effects. Protein expression profile of skin proteins is being changed with topical application of GC, however, the knowledge about singular markers in this regard is only patchy and collaboration is ill defined. Material/Methods Scar formation was observed under different doses of GC, which were locally applied on the back skin of mice (1 to 3 weeks). After euthanasia we analyzed protein expression of collagen I and III (picrosirius) in scar tissue together with 16 additional protein markers, which are involved in wound healing, with immunhistochemistry. For assessing GC's effect on co-expression we compared our results with a model of random figures to estimate how many significant correlations should be expected by chance. Results GC altered collagen and protein expression with distinct results in different areas of investigation. Most often we observed a reduced expression after application of low dose GC. In the scar infiltrate a multivariate analysis confirmed the significant impact of both GC concentrations. Calculation of Spearman's correlation coefficient similarly resulted in a significant impact of GC, and furthermore, offered the possibility to grasp the entire interactive profile in between all variables studied. The biological markers, which were connected by significant correlations could be arranged in a highly cross-linked network that involved most of the markers measured. A marker highly cross-linked with more than 3 significant correlations was indicated by a higher variation of all its correlations to the other variables, resulting in a standard deviation of > 0.2. Conclusion In addition to immunohistochemical analysis of single protein markers multivariate analysis of co-expressions by use of correlation coefficients reveals the complexity of biological relationships and identifies complex biological effects of GC on skin scarring. Depiction of collaborative clusters will help to understand functional pathways. The functional importance of highly cross-linked proteins will have to be proven in subsequent studies.
Published: 2010

24. An ANOCEF genomic and transcriptomic microarray study of the response to radiotherapy or to alkylating first-line chemotherapy in glioblastoma patients

Author: François M. Vallette, Olivier Chinot, Carole Colin, Yannick Marie, Ahmed Idbaih, Michel Wager, Aurélien de Reyniès, François Ducray, Dominique Figarella-Branger, Marc Sanson, Emilie Thomas, Jérôme Honnorat, Jean-Yves Delattre, Lucie Karayan-Tapon, David S. Rickman, François Berger, Hervé Chneiweiss, Neuro-oncologie et neuro-inflammation, Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale (INSERM), Biologie des Interactions Neurones / Glie, Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Association des Neuro-Oncologues d'Expression Française (ANOCEF), Hôpital de la Timone [CHU - APHM] (TIMONE)-CHU Marseille, (le programme) Cartes d'identité des tumeurs (CIT), Ligue Nationales Contre le Cancer (LNCC), Neuro-Oncologie [Hôpital de la Timone - APHM], Aix Marseille Université (AMU)-Assistance Publique - Hôpitaux de Marseille (APHM)- Hôpital de la Timone [CHU - APHM] (TIMONE), Centre de Recherches en Oncologie biologique et Oncopharmacologie (CRO2), Institut National de la Santé et de la Recherche Médicale (INSERM)- Hôpital de la Timone [CHU - APHM] (TIMONE)-Aix Marseille Université (AMU), Cellules souches leucémiques et thérapeuthiques, Université de Poitiers-Centre hospitalier universitaire de Poitiers (CHU Poitiers), Institut de psychiatrie et neurosciences (U894 / UMS 1266), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Recherche en Cancérologie Nantes-Angers (CRCNA), Centre Hospitalier Universitaire d'Angers (CHU Angers), PRES Université Nantes Angers Le Mans (UNAM)-PRES Université Nantes Angers Le Mans (UNAM)-Hôtel-Dieu de Nantes-Institut National de la Santé et de la Recherche Médicale (INSERM)-Hôpital Laennec-Centre National de la Recherche Scientifique (CNRS)-Faculté de Médecine d'Angers-Centre hospitalier universitaire de Nantes (CHU Nantes), Grenoble Institut des Neurosciences (GIN), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM), This work is part of the national program Cartes d'Identité des Tumeurs (CIT) funded and developed by the Ligue Nationale Contre le Cancer., Aix Marseille Université (AMU)- Hôpital de la Timone [CHU - APHM] (TIMONE)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre de Psychiatrie et Neurosciences (U894), Université Claude Bernard Lyon 1 ( UCBL ), Université de Lyon-Université de Lyon-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Association des Neuro-Oncologues d'Expression Française ( ANOCEF ), Hôpital de la Timone [CHU - APHM] ( TIMONE ) -CHU Marseille, Programme Cartes d'Identité des Tumeurs (CIT), Ligue Nationale Contre le Cancer, Aix Marseille Université ( AMU ) -Assistance Publique - Hôpitaux de Marseille ( APHM ) - Hôpital de la Timone [CHU - APHM] ( TIMONE ), Centre de Recherches en Oncologie biologique et Oncopharmacologie ( CRO2 ), Aix Marseille Université ( AMU ) - Hôpital de la Timone [CHU - APHM] ( TIMONE ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université de Poitiers-Centre hospitalier universitaire de Poitiers ( CHU Poitiers ), Centre de Psychiatrie et Neurosciences ( CPN - U894 ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Paris Descartes - Paris 5 ( UPD5 ), Centre de Recherche en Cancérologie / Nantes - Angers ( CRCNA ), CHU Angers-Hôtel-Dieu de Nantes-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Hôpital Laennec-Centre National de la Recherche Scientifique ( CNRS ) -Faculté de Médecine d'Angers-Centre hospitalier universitaire de Nantes ( CHU Nantes ), Grenoble Institut des Neurosciences ( GIN ), Université Joseph Fourier - Grenoble 1 ( UJF ) -CHU Grenoble-Institut National de la Santé et de la Recherche Médicale ( INSERM ), and BMC, Ed.
Subjects: Cancer Research, Microarray, MESH : Retrospective Studies, medicine.medical_treatment, MESH : Immunohistochemistry, MESH : Promoter Regions, Genetic, MESH: Antineoplastic Agents, Alkylating, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, Transcriptome, 0302 clinical medicine, MESH: DNA Methylation, Gene expression, Promoter Regions, Genetic, Oligonucleotide Array Sequence Analysis, 0303 health sciences, CD68, MESH: Glioblastoma, MESH : Cyclin-Dependent Kinase Inhibitor p16, lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens, Immunohistochemistry, 3. Good health, MESH : Oligonucleotide Array Sequence Analysis, Oncology, [ SDV.BBM.GTP ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], 030220 oncology & carcinogenesis, MESH: Cyclin-Dependent Kinase Inhibitor p16, DNA methylation, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Molecular Medicine, MESH : Antineoplastic Agents, Alkylating, [SDV.CAN]Life Sciences [q-bio]/Cancer, Biology, In Vitro Techniques, lcsh:RC254-282, MESH : Glioblastoma, 03 medical and health sciences, [SDV.CAN] Life Sciences [q-bio]/Cancer, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], MESH: Promoter Regions, Genetic, medicine, Humans, Antineoplastic Agents, Alkylating, Cyclin-Dependent Kinase Inhibitor p16, 030304 developmental biology, Retrospective Studies, Chemotherapy, MESH: Humans, Research, MESH : Humans, MESH: Immunohistochemistry, MESH: Retrospective Studies, DNA Methylation, Radiation therapy, MESH: Oligonucleotide Array Sequence Analysis, Cancer research, Glioblastoma, MESH : DNA Methylation
Abstract: Background The molecular characteristics associated with the response to treatment in glioblastomas (GBMs) remain largely unknown. We performed a retrospective study to assess the genomic characteristics associated with the response of GBMs to either first-line chemotherapy or radiation therapy. The gene expression (n = 56) and genomic profiles (n = 67) of responders and non-responders to first-line chemotherapy or radiation therapy alone were compared on Affymetrix Plus 2 gene expression arrays and BAC CGH arrays. Results According to Verhaak et al.'s classification system, mesenchymal GBMs were more likely to respond to radiotherapy than to first-line chemotherapy, whereas classical GBMs were more likely to respond to first-line chemotherapy than to radiotherapy. In patients treated with radiation therapy alone, the response was associated with differential expression of microenvironment-associated genes; the expression of hypoxia-related genes was associated with short-term progression-free survival (< 5 months), whereas the expression of immune genes was associated with prolonged progression-free survival (> 10 months). Consistently, infiltration of the tumor by both CD3 and CD68 cells was significantly more frequent in responders to radiotherapy than in non-responders. In patients treated with first-line chemotherapy, the expression of stem-cell genes was associated with resistance to chemotherapy, and there was a significant association between response to treatment and p16 locus deletions. Consistently, in an independent data set of patients treated with either radiotherapy alone or with both radiotherapy and adjuvant chemotherapy, we found that patients with the p16 deletion benefited from adjuvant chemotherapy regardless of their MGMT promoter methylation status, whereas in patients without the p16 deletion, this benefit was only observed in patients with a methylated MGMT promoter. Conclusion Differential expression of microenvironment genes and p16 locus deletion are associated with responses to radiation therapy and to first-line chemotherapy, respectively, in GBM. Recently identified transcriptomic subgroups of GBMs seem to respond differently to radiotherapy and to first-line chemotherapy.
Published: 2010

25. Myenteric plexus alterations downstream from a prenatal intestinal obstruction in a rat model

Author: Yoanne Mousseau, Francis Jaubert, Laurent Fourcade, Franck Sturtz, Biomolécules Thérapies anti-tumorales ( EA4021 ), Université de Limoges ( UNILIM ) -Génomique, Environnement, Immunité, Santé, Thérapeutique ( GEIST FR CNRS 3503 ), Service de chirurgie pédiatrique viscérale, orthopédique et plastique [CHU Limoges], CHU Limoges, Université de Limoges ( UNILIM ), CHU Necker - Enfants Malades [AP-HP], Service de Biochimie et Génétique Moléculaire [CHU Limoges], Biomolécules Thérapies anti-tumorales (EA4021), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503), Université de Limoges (UNILIM), and Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)
Subjects: Pathology, MESH: Intestine, Small, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH : Immunohistochemistry, MESH : Actins, MESH: Muscle, Smooth, MESH : Intestinal Obstruction, MESH : Intestinal Atresia, 0302 clinical medicine, MESH: Pregnancy, Pregnancy, Intestine, Small, MESH : Myenteric Plexus, MESH: Animals, MESH : Female, [ SDV.GEN.GH ] Life Sciences [q-bio]/Genetics/Human genetics, Myenteric plexus, 0303 health sciences, MESH : Intestine, Small, MESH : Rats, General Neuroscience, MESH : Synaptophysin, Intestinal atresia, Immunohistochemistry, Fetal Diseases, MESH: Fetal Diseases, MESH: Pregnancy Complications, [ SDV.BBM.GTP ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], [ SDV.NEU.NB ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, 030211 gastroenterology & hepatology, Female, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], medicine.medical_specialty, MESH: Rats, Intestinal Atresia, Synaptophysin, MESH : Muscle, Smooth, Motility, Myenteric Plexus, [ SDV.BBM.BM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, Biology, MESH: Actins, MESH : Pregnancy Complications, MESH: Synaptophysin, 03 medical and health sciences, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], medicine, Animals, MESH: Myenteric Plexus, Actin, 030304 developmental biology, Fetus, MESH: Intestinal Atresia, MESH : Fetal Diseases, Muscle, Smooth, MESH: Immunohistochemistry, [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Molecular biology, medicine.disease, Actins, Staining, Rats, Pregnancy Complications, MESH : Pregnancy, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, [ SDV.NEU ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], biology.protein, Enteric nervous system, MESH : Animals, MESH: Intestinal Obstruction, MESH: Female, Intestinal Obstruction
Abstract: International audience; The enteric nervous system maturation occurs during embryonic life and continues after birth. Some prenatal events on the digestive tract such as intestinal atresia have been shown to dramatically alter this maturation. Thus, we developed a fetal rat model of intestinal atresia by surgically obstructing the small bowel at embryonic day E18. Fetuses were removed at day E21, and small bowels sections were examined by immuno-histochemistry. Synaptophysin and smooth muscle actin staining was used to define the cellular aspect. Labeling revealed marked alterations of the myenteric plexus in the lower extremity of the occluded small bowel. At day E21, the myenteric plexus of the lower part and the 2 muscle layers surrounding it, retained the staining pattern observed at day E17. This cellular pattern was classified as: immature (aspect at day E17) vs. mature (aspect of day E21) by 3 pathologists not familiar with the study. The number of samples with an immature cellular pattern at the lower end of the occluded bowel was different from that observed for the upper end (Mac Nemar test, p
Published: 2009

26. Carbon dioxide effects on olfactory functioning: behavioral, histological and immunohistochemical measurements

Author: Grégory Pourié, G. Buron, Romain Hacquemand, Gérard Brand, Laboratoire de Neurosciences Intégratives et Cliniques - UFC ( NEURO ), Université Bourgogne Franche-Comté ( UBFC ) -Université de Franche-Comté ( UFC ), Laboratoire de Neurosciences Intégratives et Cliniques - UFC (EA 481) (NEURO), Université de Franche-Comté (UFC), and Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)
Subjects: Pathology, MESH : Olfactory Marker Protein, MESH: Olfactory Perception, MESH: Air Pollutants, MESH : Immunohistochemistry, MESH : Dose-Response Relationship, Drug, Toxicology, MESH: Carbon Dioxide, MESH : Behavior, Animal, Epithelium, MESH: Dose-Response Relationship, Drug, MESH : Maze Learning, Mice, 0302 clinical medicine, MESH: Behavior, Animal, MESH: Olfactory Marker Protein, MESH: Animals, MESH : Female, MESH : Olfactory Bulb, 0303 health sciences, Air Pollutants, Inhalation Exposure, MESH : Mice, Inbred Strains, biology, Inhalation, Behavior, Animal, General Medicine, Immunohistochemistry, Olfactory Bulb, MESH : Carbon Dioxide, Sensory Thresholds, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Female, MESH: Inhalation Exposure, MESH: Olfactory Bulb, medicine.medical_specialty, Period (gene), MESH : Air Pollutants, Mice, Inbred Strains, Olfaction, MESH: Mice, Inbred Strains, Andrology, 03 medical and health sciences, Olfactory Marker Protein, Proliferating Cell Nuclear Antigen, MESH : Mice, MESH : Olfactory Perception, medicine, Animals, Maze Learning, MESH: Mice, 030304 developmental biology, Dose-Response Relationship, Drug, MESH: Maze Learning, Histology, MESH: Immunohistochemistry, Carbon Dioxide, Olfactory Perception, MESH : Epithelium, Proliferating cell nuclear antigen, MESH: Epithelium, MESH: Proliferating Cell Nuclear Antigen, Odor, 13. Climate action, MESH : Proliferating Cell Nuclear Antigen, [ SDV.NEU ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], biology.protein, MESH : Animals, MESH : Inhalation Exposure, MESH: Sensory Thresholds, Olfactory marker protein, MESH: Female, 030217 neurology & neurosurgery, MESH : Sensory Thresholds
Abstract: International audience; Most studies on toxic inhalation focus on solvent effects and few have dealt with gases on olfactory functioning. Among gases, the effects of carbon dioxide on general physiology have been well investigated contrary to the impact on olfactory neuroepithelium. Thus, this work was designed to evaluate in mice the possible effects of 3% CO(2) in two exposure periods: a 5h/day and a 12h/day conditions. Behavioral, histological and immunohistochemical observations were conducted every 2 weeks, i.e. before (W0), during (W2, W4) and after exposure (W6, W8). Firstly, behavioral evaluations of odor sensitivity showed differences in relation to the odor tested, i.e. no effect with congener urine odor and a reinforcement of 2,4,5-trimethythiazoline (TMT) (predator odor) repulsion. Secondly, histological evaluations showed a similar evolution of the epithelium thickness, i.e. a decrease along the exposure as well as during the post-exposure period and an increase of cell number (whatever the phenotype) although the kinetic appeared different in both experimental conditions. Thirdly, immunohistochemical quantification of olfactory marker protein (OMP)- and proliferating cell nuclear antigen (PCNA)-positive cells revealed that the number of mature olfactory neurons increased at the early beginning of exposure period in both conditions. While a decrease was observed in the following weeks (W4-W8) for the 12h/day condition, a stable amount of OMP-positive cells was maintained in the 5h/day condition. In contrast, the number of PCNA-positive cells followed a similar evolution, i.e. a constant decrease along the experiment. These findings indicate that the effects of CO(2) inhalation exposure are selectively dose-dependent.
Published: 2009

27. Role of regulatory T cells in a new mouse model of experimental autoimmune myositis

Author: Yves Allenbach, Benoît L. Salomon, Odile Dubourg, S. Solly, Serge Herson, Lucile Musset, Olivier Benveniste, Sylvie Grégoire, David Klatzmann, Gillian Butler-Browne, Facultat de Química (DEPARTAMENT DE QUíMICA ORGàNICA), Universitat de Barcelona (UB), Laboratoire de Biologie et Thérapeutique des Pathologies Immunitaires (CNRS UMR 7087), Centre National de la Recherche Scientifique (CNRS), Service de cardiologie et maladies vasculaires, Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Ambroise Paré, UMR S 787, Université Pierre et Marie Curie - Paris 6 (UPMC), EA 209 - Transports membranaires et chimiorésistances (LABORATOIRE DE PARASITOLOGIE-MYCOLOGIE), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Université Paris Descartes - Paris 5 (UPD5), Service de biothérapies [CHU Pitié-Salpétrière], Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Pitié-Salpêtrière [APHP], CHU Pitié-Salpêtrière [APHP], Facultat de Química ( DEPARTAMENT DE QUíMICA ORGàNICA ), Universitat de Barcelona ( UB ), Laboratoire de Biologie et Thérapeutique des Pathologies Immunitaires ( CNRS UMR 7087 ), Centre National de la Recherche Scientifique ( CNRS ), Université de Versailles Saint-Quentin-en-Yvelines ( UVSQ ) -Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Ambroise Paré, Université Pierre et Marie Curie - Paris 6 ( UPMC ), EA 209 - Transports membranaires et chimiorésistances ( LABORATOIRE DE PARASITOLOGIE-MYCOLOGIE ), Assistance publique - Hôpitaux de Paris (AP-HP)-Université Paris Descartes - Paris 5 ( UPD5 ), Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Pitié-Salpêtrière [APHP], Service de cardiologie et maladies vasculaires [CHU Ambroise Paré], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Ambroise Paré [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Descartes - Paris 5 (UPD5), Departement Hospitalo- Universitaire - Inflammation, Immunopathologie, Biothérapie [Paris] (DHU - I2B), Sorbonne Université (SU)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-CHU Trousseau [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Biologie et thérapeutique des pathologies immunitaires (BTPI), and Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)
Subjects: MESH: Inflammation, CD4-Positive T-Lymphocytes, MESH : Immunohistochemistry, MESH: Interleukin-2 Receptor alpha Subunit, medicine.disease_cause, Polymyositis, T-Lymphocytes, Regulatory, Autoimmunity, Nervous System Autoimmune Disease, Experimental, MESH : T-Lymphocytes, Regulatory, Mice, 0302 clinical medicine, MESH: Autoimmune Diseases, MESH : Female, MESH: Animals, IL-2 receptor, MESH : Muscle, Skeletal, Myositis, 0303 health sciences, education.field_of_study, MESH: Muscle, Skeletal, B-Lymphocytes, Mice, Inbred BALB C, MESH: Muscle Strength, MESH: CD4-Positive T-Lymphocytes, MESH : Interleukin-2 Receptor alpha Subunit, Immunohistochemistry, 3. Good health, MESH : CD4-Positive T-Lymphocytes, MESH: Immunization, Female, medicine.symptom, MESH : Nervous System Autoimmune Disease, Experimental, Population, MESH: Mice, Inbred BALB C, Inflammation, [ SDV.IMM.IMM ] Life Sciences [q-bio]/Immunology/Immunotherapy, MESH : Myosins, Muscle disorder, Biology, Myosins, Pathology and Forensic Medicine, MESH : B-Lymphocytes, Autoimmune Diseases, MESH: Nervous System Autoimmune Disease, Experimental, 03 medical and health sciences, MESH : Autoimmune Diseases, MESH: B-Lymphocytes, MESH : Mice, medicine, Animals, Muscle Strength, education, Muscle, Skeletal, MESH: Mice, MESH : Mice, Inbred BALB C, 030304 developmental biology, MESH : Inflammation, MESH: T-Lymphocytes, Regulatory, Interleukin-2 Receptor alpha Subunit, MESH: Immunohistochemistry, MESH : Immunization, [SDV.IMM.IMM]Life Sciences [q-bio]/Immunology/Immunotherapy, MESH: Myosins, medicine.disease, MESH : Muscle Strength, Immunology, Immunization, MESH : Animals, MESH: Female, CD8, 030215 immunology, Regular Articles
Abstract: International audience; Polymyositis is a rare and severe inflammatory muscle disorder. Treatments are partially efficacious but have many side effects. New therapeutic approaches must be first tested in a relevant animal model. Regulatory CD4+CD25+ T cells (Tregs) have been rediscovered as a pivotal cell population in the control of autoimmunity, but the connection between polymyositis and Tregs is currently unknown. To develop a reproducible experimental autoimmune myositis model of polymyositis, mice were immunized once a week for 3 weeks with 1 mg of partially purified myosin emulsified in complete Freund's adjuvant. All mice injected with myosin and complete Freund's adjuvant developed myositis. The infiltrates were composed of CD4(+) and CD8(+) cells, as well as macrophages, but did not contain B lymphocytes. In mice that were depleted of Tregs, the myositis was more severe, as determined by quantitative scoring of muscle inflammation (2.36 +/- 0.9 vs. 1.64 +/- 0.8, P = 0.019). In contrast, injection of in vitro expanded polyclonal Tregs at the time of immunization significantly improved the disease (quantitative score of inflammation 0.87 +/- 1.06 vs. 2.4 +/- 0.67, P = 0.047). Transfer of sensitized or CD4(+)-sorted cells from the lymph nodes of experimental autoimmune myositis mice induced myositis in naïve, irradiated, recipient mice. Thus, experimental autoimmune myositis is a reproducible, transferable disease in mice, both aggravated by Treg depletion and improved by polyclonal Treg injection.
Published: 2009

28. The expression of Twist has an impact on survival in human bladder cancer and is influenced by the smoking status

Author: Jean P. Thiery, Hugues Bittard, Marie E. Fondrevelle, Franck Monnien, Hervé Wallerand, Robert E. Reiter, Dominique Chopin, Bernadette Kantelip, Laboratoire d'anatomie pathologique [Besancon], Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Hôpital Jean Minjoz-Université de Franche-Comté ( UFC ), Carcinogénèse épithéliale : facteurs prédictifs et pronostiques - UFC ( CEF2P / CARCINO ), Université Bourgogne Franche-Comté ( UBFC ) -Université de Franche-Comté ( UFC ) -Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ), Service de Néphrologie et Urologie, Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Hôpital Saint-Jacques, Service d'urologie, andrologie et transplantation rénale, Université Bordeaux Segalen - Bordeaux 2-CHU Bordeaux [Bordeaux]-Groupe hospitalier Pellegrin, Service d'Anatomie pathologique [CHRU Besançon], Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon), Carcinogénèse épithéliale : facteurs prédictifs et pronostiques - UFC (EA 3181) (CEF2P / CARCINO), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), and Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Hôpital Saint-Jacques
Subjects: Oncology, Male, Pathology, MESH : Immunohistochemistry, MESH : Aged, Kaplan-Meier Estimate, MESH: Cadherins, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, MESH: Proportional Hazards Models, 0302 clinical medicine, MESH: Aged, 80 and over, MESH: Risk Factors, Risk Factors, MESH : Neoplasm Staging, MESH : Female, MESH: Kaplan-Meiers Estimate, Stage (cooking), MESH: Aged, Aged, 80 and over, 0303 health sciences, MESH: Middle Aged, MESH : Prognosis, Smoking, MESH: Twist Transcription Factor, Nuclear Proteins, Anatomical pathology, MESH: Neoplasm Staging, Middle Aged, Cadherins, Prognosis, Immunohistochemistry, MESH : Risk Factors, 3. Good health, MESH: Urinary Bladder Neoplasms, MESH : Smoking, MESH : Urinary Bladder Neoplasms, 030220 oncology & carcinogenesis, Disease Progression, MESH: Disease Progression, Female, MESH : Cadherins, medicine.medical_specialty, MESH: Smoking, Urology, MESH : Male, [SDV.CAN]Life Sciences [q-bio]/Cancer, MESH: Multivariate Analysis, MESH: Prognosis, 03 medical and health sciences, Internal medicine, medicine, Humans, MESH : Middle Aged, Epithelial–mesenchymal transition, Risk factor, MESH : Aged, 80 and over, 030304 developmental biology, Aged, Neoplasm Staging, Proportional Hazards Models, Bladder cancer, MESH: Humans, Proportional hazards model, business.industry, Twist-Related Protein 1, MESH : Twist Transcription Factor, MESH : Humans, Cancer, MESH : Multivariate Analysis, MESH : Nuclear Proteins, MESH: Immunohistochemistry, MESH : Kaplan-Meiers Estimate, MESH : Disease Progression, medicine.disease, MESH : Proportional Hazards Models, MESH: Male, Urinary Bladder Neoplasms, Multivariate Analysis, business, MESH: Nuclear Proteins, MESH: Female
Abstract: International audience; OBJECTIVES: Twist is considered as transcription factor that regulates epithelial mesenchymal transition (EMT) by at least inhibition of E-cadherin expression. EMT is a key event in the tumor invasion process. The purpose of this study is to investigate the expression of Twist but also those of E- and N-cadherin in human primary bladder tumor and to evaluate its prognostic value. As smoking cigarettes is a strong bladder cancer risk factor, we tried to evaluate the impact of the tobacco status on these molecular abnormalities. MATERIALS AND METHODS: To delineate on the oncogenic role for Twist in human bladder cancer, we evaluated the E- and N-cadherin but also Twist expression (n = 70) by immunohistochemistry. We evaluated the prognostic value of these expressions. Moreover, we tried to correlate these protein expressions to the smoking status of the patients. Overall survival (OS) and progression-free survival (PFS) were evaluated using the Kaplan-Meier method, and multivariate analysis was performed using the Cox proportional hazard analysis. RESULTS: Of the 70 bladder tumors, 28 (40%) cases were positive for Twist expression, 16 (23%) cases were negative for E-cadherin expression, and 12 (17%) were positive for N-cadherin expression. When categorized into negative vs. positive expression, Twist was associated with the stage (P = 0.001), the grade (P < 0.001), the progression (P = 0.02), and the E-cadherin expression (P = 0.01). Moreover, positive Twist expression clearly predicted poorer PFS (P = 0.02). In the multivariate analysis, both positive Twist expression and loss of E-cadherin expression were independent prognostic factors for PFS (P = 0.046 and P = 0.001, respectively) and only loss of E-cadherin expression for the OS (P < 0.001). We also demonstrated that almost 60% (16/28) of patients with Twist-positive expression were current smokers at the time of the diagnosis, corroborating the fact that smoking modulates the expression of EMT markers including Twist. CONCLUSION: Positive Twist expression may be a useful prognostic marker for patients with bladder cancer. Its expression seems to be correlated to the tobacco status of the patients.
Published: 2009

29. The CDK4-pRB-E2F1 pathway controls insulin secretion

Author: Stéphane Dalle, Lluis Fajas, Safia Costes, Emilie Blanchet, Said Assou, Jean-Sébastien Annicotte, Irena Iankova, Claude Sardet, Carine Chavey, Jacques Teyssier, INSERM, U834, Montpellier, F-34298, France, Métabolisme et cancer, Université Montpellier 1 ( UM1 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), CRLC Val d'Aurelle-Paul Lamarque, CRLCC Val d'Aurelle - Paul Lamarque, Institut de recherche en cancérologie de Montpellier ( IRCM ), Université Montpellier 1 ( UM1 ) -CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université de Montpellier ( UM ), Institut de Génomique Fonctionnelle ( IGF ), Centre National de la Recherche Scientifique ( CNRS ) -Université Montpellier 2 - Sciences et Techniques ( UM2 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Montpellier 1 ( UM1 ) -Université de Montpellier ( UM ), Institut de recherche en biothérapie ( IRB ), Université Montpellier 1 ( UM1 ) -Centre Hospitalier Régional Universitaire [Montpellier] ( CHRU Montpellier ) -Université de Montpellier ( UM ), Institut de Génétique Moléculaire de Montpellier ( IGMM ), Université de Montpellier ( UM ) -Centre National de la Recherche Scientifique ( CNRS ), CHU Arnaud de Villeuneuve Montpellier, Centre Hospitalier Régional Universitaire [Montpellier] ( CHRU Montpellier ), INSERM-Association Française des Diabétiques (PNR-Diabète), Association pour la Recherche contre le Cancer, and Fondation pour la Recherche Médicale. E.B. is supported by a grant form the Ministère de l'Enseignement Supérieur et de la Recherche, C.C. is supported by a grant from the Agence Nationale pour la Recherche., Agence Nationale pour la Recherche (ANR physio2006),Agence Nationale pour la Recherche (ANR physio2006), Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de recherche en cancérologie de Montpellier (IRCM - U896 Inserm - UM1), Université Montpellier 1 (UM1)-CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), Institut de Génomique Fonctionnelle (IGF), Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut de recherche en biothérapie (IRB), Université Montpellier 1 (UM1)-Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), Institut de Génétique Moléculaire de Montpellier (IGMM), Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), Agence Nationale pour la Recherche (ANR physio2006), Le Ster, Yves, CRLCC Val d'Aurelle - Paul Lamarque-Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 1 (UM1), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM)
Subjects: [SDV.MHEP.PHY] Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], medicine.medical_treatment, MESH : Blotting, Western, MESH : RNA, Small Interfering, MESH : E2F1 Transcription Factor, MESH : Phosphorylation, Mice, 0302 clinical medicine, Insulin Secretion, Glucose homeostasis, Insulin, MESH: Animals, ComputingMilieux_MISCELLANEOUS, MESH: Chromatin Immunoprecipitation, 0303 health sciences, Retinoblastoma protein, MESH : Reverse Transcriptase Polymerase Chain Reaction, MESH : Protein Binding, Transfection, [SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, [ SDV.MHEP.EM ] Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, Immunohistochemistry, Cell biology, MESH: COS Cells, [ SDV.BBM.GTP ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], 030220 oncology & carcinogenesis, COS Cells, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], endocrine system, Blotting, Western, MESH: Insulin, Article, 03 medical and health sciences, MESH: Gene Expression Profiling, Islets of Langerhans, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], MESH: Blotting, Western, MESH: Protein Binding, MESH : Islets of Langerhans, Potassium Channels, Inwardly Rectifying, neoplasms, MESH: Phosphorylation, MESH : Glucose, MESH: Islets of Langerhans, MESH : Gene Expression Profiling, MESH: Cercopithecus aethiops, MESH: Cell Line, Mice, Inbred C57BL, Insulin receptor, Glucose, MESH : Cercopithecus aethiops, Chromatin immunoprecipitation, MESH: Signal Transduction, MESH : Cell Line, MESH : Immunohistochemistry, MESH : Insulin, [SDV]Life Sciences [q-bio], MESH : Models, Biological, MESH: Mice, Knockout, Retinoblastoma Protein, MESH: Reverse Transcriptase Polymerase Chain Reaction, MESH: RNA, Small Interfering, Chlorocebus aethiops, Phosphorylation, RNA, Small Interfering, [SDV.MHEP.EM] Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism, Mice, Knockout, biology, [ SDV.MHEP.PHY ] Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], Reverse Transcriptase Polymerase Chain Reaction, MESH: Potassium Channels, Inwardly Rectifying, MESH: E2F1 Transcription Factor, MESH: Glucose, MESH : Cyclin-Dependent Kinase 4, Regulatory Pathway, Signal transduction, MESH : Transfection, biological phenomena, cell phenomena, and immunity, MESH : COS Cells, Protein Binding, Signal Transduction, Chromatin Immunoprecipitation, MESH: Cyclin-Dependent Kinase 4, MESH : Mice, Inbred C57BL, Models, Biological, Cell Line, MESH : Chromatin Immunoprecipitation, MESH: Mice, Inbred C57BL, MESH : Mice, medicine, [SDV.MHEP.PHY]Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO], Animals, MESH: Mice, 030304 developmental biology, MESH : Signal Transduction, MESH: Transfection, Gene Expression Profiling, MESH: Models, Biological, Cyclin-Dependent Kinase 4, MESH: Immunohistochemistry, Cell Biology, MESH: Retinoblastoma Protein, MESH : Potassium Channels, Inwardly Rectifying, biology.protein, MESH : Mice, Knockout, MESH : Animals, MESH : Retinoblastoma Protein, E2F1 Transcription Factor
Abstract: International audience; CDK4-pRB-E2F1 cell-cycle regulators are robustly expressed in non-proliferating beta cells, suggesting that besides the control of beta-cell number the CDK4-pRB-E2F1 pathway has a role in beta-cell function. We show here that E2F1 directly regulates expression of Kir6.2, which is a key component of the K(ATP) channel involved in the regulation of glucose-induced insulin secretion. We demonstrate, through chromatin immunoprecipitation analysis from tissues, that Kir6.2 expression is regulated at the promoter level by the CDK4-pRB-E2F1 pathway. Consistently, inhibition of CDK4, or genetic inactivation of E2F1, results in decreased expression of Kir6.2, impaired insulin secretion and glucose intolerance in mice. Furthermore we show that rescue of Kir6.2 expression restores insulin secretion in E2f1(-/-) beta cells. Finally, we demonstrate that CDK4 is activated by glucose through the insulin pathway, ultimately resulting in E2F1 activation and, consequently, increased expression of Kir6.2. In summary we provide evidence that the CDK4-pRB-E2F1 regulatory pathway is involved in glucose homeostasis, defining a new link between cell proliferation and metabolism.
Published: 2008

30. The uterosacral complex: ligament or neurovascular pathway? Anatomical and histological study of fetuses and adults

Author: Francine Arbez-Gindre, Bernard Parratte, Rajeev Ramanah, Didier Riethmuller, Robert Maillet, Service de gynécologie, Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Hôpital Saint-Jacques, Service de médecine physique et de réadaptation, Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ), Carcinogénèse épithéliale : facteurs prédictifs et pronostiques - UFC ( CEF2P / CARCINO ), Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Université Bourgogne Franche-Comté ( UBFC ) -Université de Franche-Comté ( UFC ), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Hôpital Saint-Jacques, Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon), Carcinogénèse épithéliale : facteurs prédictifs et pronostiques - UFC (EA 3181) (CEF2P / CARCINO), Université de Franche-Comté (UFC), and Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)
Subjects: Pathology, Biopsy, MESH : Immunohistochemistry, Uterosacral ligament, Lumbosacral Plexus, MESH: Hypogastric Plexus, H&E stain, MESH : Aged, MESH: Ligaments, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, MESH: Biopsy, 0302 clinical medicine, MESH: Aged, 80 and over, MESH: Pregnancy, Pregnancy, MESH : Uterus, MESH : Female, Aged, 80 and over, MESH: Aged, 030219 obstetrics & reproductive medicine, Pelvic floor, MESH: Lumbosacral Plexus, medicine.diagnostic_test, MESH : Hypogastric Plexus, Obstetrics and Gynecology, Hypogastric Plexus, Anatomy, MESH : Fetus, Immunohistochemistry, 3. Good health, medicine.anatomical_structure, MESH: Young Adult, 030220 oncology & carcinogenesis, MESH: Sacrococcygeal Region, MESH : Cadaver, Ligament, MESH: Uterus, MESH : Lumbosacral Plexus, Female, medicine.medical_specialty, Urology, MESH : Young Adult, Connective tissue, [SDV.CAN]Life Sciences [q-bio]/Cancer, MESH : Blood Vessels, 03 medical and health sciences, Young Adult, Fetus, medicine, Cadaver, Humans, MESH: Cadaver, MESH : Aged, 80 and over, Aged, Ligaments, MESH: Humans, business.industry, Sacrococcygeal Region, Uterus, MESH : Humans, MESH: Fetus, MESH: Immunohistochemistry, MESH: Blood Vessels, Neurovascular bundle, MESH : Pregnancy, MESH : Biopsy, MESH : Sacrococcygeal Region, Blood Vessels, business, MESH : Ligaments, MESH: Female
Abstract: International audience; The aim of this study was to define the anatomical relationships of the uterosacral ligament complex (USLC) and to analyze histologically its content. Three fetal and four adult cadavers were used. Anatomical dissections were carried out. Eight fresh biopsies (four fetal and four adult) of the USLC were analyzed histologically and immunohistochemically. Specimens were stained with hematoxylin eosin safran coloration, with anti-nervous cell antibodies (PS 100) and with anti-smooth muscle antibodies (to visualize vessel walls). By removing the visceral pelvic fascia, nervous fibers were found within the USLC forming the hypogastric plexus. Histologically, the USLC contained connective tissue, nervous fibers, sympathetic nodes, vessels, and fatty tissue. No structured ligamentous organization was identified. The uterosacral "ligament" is a "complex" integrating connective tissue as well as nervous and vascular elements. Radical excisions and USLC suspension during pelvic floor reconstructive surgery should be performed with caution in order to preserve pelvic innervation.
Published: 2008

31. Phase III trial comparing intensive induction chemoradiotherapy (60 Gy, infusional 5-FU and intermittent cisplatin) followed by maintenance gemcitabine with gemcitabine alone for locally advanced unresectable pancreatic cancer. Definitive results of the 2000-01 FFCD/SFRO study

Author: Ph. Rougier, Thomas Aparicio, Christophe Mariette, F. Mornex, J.-F. Bosset, O. Bouche, Franck Bonnetain, Laurent Mineur, A. Azzedine, Pascal Hammel, Bruno Chauffert, N. Stremsdoerfer, J. Butel, Philippe Maingon, Laurent Bedenne, Lipides - Nutrition - Cancer (U866) ( LNC ), Université de Bourgogne ( UB ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon ( ENSBANA ), Grand Accélérateur National d'Ions Lourds ( GANIL ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ) -Institut National de Physique Nucléaire et de Physique des Particules du CNRS ( IN2P3 ) -Centre National de la Recherche Scientifique ( CNRS ), Carcinogénèse épithéliale : facteurs prédictifs et pronostiques - UFC ( CEF2P / CARCINO ), Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Université Bourgogne Franche-Comté ( UBFC ) -Université de Franche-Comté ( UFC ), Lipides - Nutrition - Cancer (U866) (LNC), Université de Bourgogne (UB)-Institut National de la Santé et de la Recherche Médicale (INSERM)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Ecole Nationale Supérieure de Biologie Appliquée à la Nutrition et à l'Alimentation de Dijon (ENSBANA), Grand Accélérateur National d'Ions Lourds (GANIL), Centre National de la Recherche Scientifique (CNRS)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3), Carcinogénèse épithéliale : facteurs prédictifs et pronostiques - UFC (EA 3181) (CEF2P / CARCINO), Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon), Matrice extracellulaire et dynamique cellulaire - UMR 7369 (MEDyC), Université de Reims Champagne-Ardenne (URCA)-SFR CAP Santé (Champagne-Ardenne Picardie Santé), and Université de Reims Champagne-Ardenne (URCA)-Université de Picardie Jules Verne (UPJV)-Université de Reims Champagne-Ardenne (URCA)-Université de Picardie Jules Verne (UPJV)-Centre National de la Recherche Scientifique (CNRS)
Subjects: Male, MESH: Remission Induction, MESH : Antineoplastic Combined Chemotherapy Protocols, medicine.medical_treatment, MESH : Aged, MESH: Risk Assessment, Deoxycytidine, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, 0302 clinical medicine, Antineoplastic Combined Chemotherapy Protocols, MESH : Neoplasm Staging, ComputingMilieux_MISCELLANEOUS, MESH: Treatment Outcome, MESH: Middle Aged, Biopsy, Needle, Remission Induction, MESH : Infusions, Intravenous, MESH : Chemotherapy, Adjuvant, MESH: Follow-Up Studies, Immunohistochemistry, 3. Good health, MESH: Antineoplastic Combined Chemotherapy Protocols, Oncology, Fluorouracil, 030220 oncology & carcinogenesis, 030211 gastroenterology & hepatology, medicine.medical_specialty, MESH: Radiotherapy, Adjuvant, MESH: Probability, MESH : Drug Administration Schedule, Urology, MESH: Drug Administration Schedule, Antimetabolite, Risk Assessment, Drug Administration Schedule, MESH : Radiotherapy, Adjuvant, MESH : Cisplatin, 03 medical and health sciences, Humans, MESH : Middle Aged, Aged, MESH: Humans, Dose-Response Relationship, Drug, MESH: Deoxycytidine, MESH : Humans, MESH: Adult, MESH : Follow-Up Studies, Survival Analysis, Pancreatic Neoplasms, Concomitant, Cisplatin, MESH: Female, MESH: Fluorouracil, MESH : Pancreatic Neoplasms, MESH: Combined Modality Therapy, MESH : Immunohistochemistry, MESH : Dose-Response Relationship, Drug, MESH : Biopsy, Needle, MESH : Neoplasm Invasiveness, MESH: Dose-Response Relationship, Drug, MESH : Female, MESH : Risk Assessment, MESH: Radiotherapy Dosage, Infusions, Intravenous, MESH: Aged, MESH : Radiotherapy Dosage, Radiotherapy Dosage, Hematology, MESH: Neoplasm Staging, Middle Aged, MESH : Adult, Chemotherapy regimen, Combined Modality Therapy, Treatment Outcome, MESH: Chemotherapy, Adjuvant, Chemotherapy, Adjuvant, MESH: Survival Analysis, MESH : Fluorouracil, Female, MESH: Pancreatic Neoplasms, medicine.drug, Adult, MESH: Biopsy, Needle, medicine.drug_class, MESH : Male, MESH : Probability, [SDV.CAN]Life Sciences [q-bio]/Cancer, MESH : Treatment Outcome, MESH : Deoxycytidine, medicine, Neoplasm Invasiveness, MESH: Infusions, Intravenous, Neoplasm Staging, Probability, Chemotherapy, MESH : Remission Induction, Performance status, business.industry, MESH: Immunohistochemistry, MESH: Neoplasm Invasiveness, Gemcitabine, MESH: Male, Surgery, MESH: Cisplatin, Radiotherapy, Adjuvant, MESH : Survival Analysis, business, MESH : Combined Modality Therapy, Chemoradiotherapy, Follow-Up Studies
Abstract: International audience; BACKGROUND: The role of chemoradiation with systemic chemotherapy compared with chemotherapy alone in locally advanced pancreatic cancer (LAPC) is uncertain. PATIENTS AND METHODS: One hundred and nineteen patients with LAPC, World Health Organization performance status of zero to two were randomly assigned to either the induction CHRT group (60 Gy, 2 Gy/fraction; concomitant 5-fluorouracil infusion, 300 mg/m(2)/day, days 1-5 for 6 weeks; cisplatin, 20 mg/m(2)/day, days 1-5 during weeks 1 and 5) or the induction gemcitabine group (GEM: 1000 mg/m(2) weekly for 7 weeks). Maintenance gemcitabine (1000 mg/m(2) weekly, 3/4 weeks) was given in both arms until disease progression or toxicity. RESULTS: Overall survival was shorter in the CHRT than in GEM arm [median survival 8.6 (99% confidence interval 7.1-11.4) and 13 months (8.7-18.1), P = 0.03]. One-year survival was, respectively, 32% and 53%. These results were confirmed in a per-protocol analysis for patients who received 75% or more of the planned dose of radiotherapy. More overall grades 3-4 toxic effects were recorded in the CHRT arm, both during induction (36 versus 22%) and maintenance (32 versus 18%). CONCLUSION: This intensive induction schedule of CHRT was more toxic and less effective than gemcitabine alone.
Published: 2008

32. KBA.62: a useful marker for primary and metastatic melanomas

Author: Sophie Le Guellec, Philippe Rochaix, Paul Carle, Pierre Payoux, Séverine Valmary-Degano, Talal Al Saati, François Launay, Laurence Lamant, Serge Boulinguez, Frédéric Lauwers, Pierre Brousset, Cécile Pages, Laboratoire d'Anatomie Pathologique [Purpan], CHU Toulouse [Toulouse], Institut Claudius Regaud, CRLCC Institut Claudius Regaud, Plateau technique d'Histopathologie Expérimentale, Institut Claude de Préval (ICP), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Toulouse [Toulouse]-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Toulouse [Toulouse]-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Service de dermatologie, Hôpital Purpan [Toulouse], CHU Toulouse [Toulouse]-CHU Toulouse [Toulouse], Service de chirurgie plastique et maxillo-faciale, CHU Purpan, Toulouse, Service de Médecine Nucléaire [Toulouse], CHU Toulouse [Toulouse]-Hôpital Rangueil, Carcinogénèse épithéliale : facteurs prédictifs et pronostiques - UFC ( CEF2P / CARCINO ), Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Université Bourgogne Franche-Comté ( UBFC ) -Université de Franche-Comté ( UFC ), Laboratoire d'anatomie pathologique [Besancon], Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Hôpital Jean Minjoz-Université de Franche-Comté ( UFC ), Institut de Physique Nucléaire d'Orsay ( IPNO ), Université Paris-Sud - Paris 11 ( UP11 ) -Institut National de Physique Nucléaire et de Physique des Particules du CNRS ( IN2P3 ) -Centre National de la Recherche Scientifique ( CNRS ), Imagerie cérébrale et handicaps neurologiques, Université Paul Sabatier - Toulouse 3 ( UPS ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Service d'anatomie pathologique et histologie-cytologie [Rangueil], Université Paul Sabatier - Toulouse 3 ( UPS ) -CHU Toulouse [Toulouse]-Hôpital de Rangueil, Université Sciences et Technologies - Bordeaux 1, Centre de Recherche en Cancérologie de Toulouse ( CRCT ), and Université Paul Sabatier - Toulouse 3 ( UPS ) -CHU Toulouse [Toulouse]-Hôpital Purpan [Toulouse]-Institut National de la Santé et de la Recherche Médicale ( INSERM )
Subjects: MESH: Neoplasm Proteins, Pathology, Skin Neoplasms, MESH : Immunohistochemistry, MESH: Lymphatic Metastasis, MESH : S100 Proteins, MESH: Antigens, Neoplasm, MESH: Lymph Nodes, MESH : Antigens, Neoplasm, Metastasis, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, MESH: Antibodies, Monoclonal, 030207 dermatology & venereal diseases, 0302 clinical medicine, MESH : Tumor Markers, Biological, Lymph node, Melanoma, MESH : Lymph Nodes, Micrometastasis, S100 Proteins, Antibodies, Monoclonal, MESH : Lymphatic Metastasis, Immunohistochemistry, 3. Good health, Neoplasm Proteins, medicine.anatomical_structure, 030220 oncology & carcinogenesis, MESH : Antibodies, Monoclonal, Lymphatic Metastasis, Lymph, MESH : Sensitivity and Specificity, MESH: S100 Proteins, Melanoma-Specific Antigens, medicine.medical_specialty, MESH: Melanoma, Sentinel lymph node, MESH : Melanoma, [SDV.CAN]Life Sciences [q-bio]/Cancer, Sensitivity and Specificity, Pathology and Forensic Medicine, 03 medical and health sciences, MESH : Neoplasm Proteins, Antigens, Neoplasm, medicine, Biomarkers, Tumor, Humans, Desmoplastic melanoma, MESH: Humans, business.industry, MESH: Skin Neoplasms, MESH : Skin Neoplasms, MESH : Humans, MESH : Melanoma-Specific Antigens, MESH: Immunohistochemistry, medicine.disease, MESH: Sensitivity and Specificity, MESH: Melanoma-Specific Antigens, MESH: Tumor Markers, Biological, Lymph Nodes, business
Abstract: International audience; We previously described a novel antimelanoma antibody, designated KBA.62. However, review of the literature showed that only a few studies have reported on this antibody. We report our experience in the diagnosis of melanoma using KBA.62 and its value in both primary and metastatic conditions. In addition to conventional melanomas, we included desmoplastic and spindle cell melanomas, whose diagnosis can be challenging. We also focus on identification of malignant cells in sentinel lymph node biopsies using KBA.62, by comparison with anti-S-100 protein and HMB-45 antibodies, because discrepancies in sensitivity and specificity of melanoma markers have given rise to numerous studies aiming to determine the best panel for the evaluation of sentinel lymph node from patients with melanoma. Overall, KBA.62 was positive in 93% of primary and metastatic melanomas. Interestingly, the 12 cases of desmoplastic and spindle cell melanomas had strongly positive results. In addition, the results of our study performed on a series of 215 sentinel lymph nodes showed that the sensitivity of anti-S-100 protein and KBA.62 antibodies in detecting occult metastasis was similar. Moreover, KBA.62 identified 6 patients (3%) who had confirmed sentinel lymph node metastasis but were negative for HMB-45. The resolution was higher with KBA.62 than that observed with anti-S-100 protein antibody, as the nonmelanocytic positive cells for KBA.62 in lymph nodes were only represented by endothelial cells, which therefore constituted an intrinsic positive control. We conclude that KBA.62 antibody is a useful additional marker for melanoma, specifically in desmoplastic/spindle cell cases and in the context of micrometastasis in sentinel lymph node.
Published: 2008

33. Secretory IgA mediates retrotranscytosis of intact gliadin peptides via the transferrin receptor in celiac disease.: IgA retrotranscytosis in celiac disease

Author: Matysiak-Budnik, Tamara, Cruz-Moura, Ivan, Arcos-Fajardo, Michelle, Lebreton, Corinne, Ménard, Sandrine, Candalh, Céline, Ben Khalifa, Karima, Dugave, Christophe, Tamouza, Houda, Van Niel, Guillaume, Bouhnik, Yoram, Lamarque, Dominique, Chaussade, Stanislas, Malamut, Georgia, Cellier, Christophe, Cerf-Bensussan, Nadine, Monteiro, Renato, Heyman, Martine, Interactions de l'épithelium intestinal avec le système immunitaire, Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de gastro-entérologie et assistance nutritive, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7)-Hôpital Beaujon [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Immunopathologie rénale, récepteurs et inflammation, Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Département d'Ingénierie et d'Etudes des Protéines, Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Compartimentation et dynamique cellulaires (CDC), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut Curie [Paris]-Centre National de la Recherche Scientifique (CNRS), Service d'hépato-gastro-entérologie, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Européen Georges Pompidou [APHP] (HEGP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO)-Hôpitaux Universitaires Paris Ouest - Hôpitaux Universitaires Île de France Ouest (HUPO), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Assistance publique - Hôpitaux de Paris (AP-HP)-Université Paris Diderot - Paris 7 ( UPD7 ) -Hôpital Beaujon, Université Paris Diderot - Paris 7 ( UPD7 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Commissariat à l'énergie atomique et aux énergies alternatives ( CEA ), Compartimentation et dynamique cellulaires ( CDC ), Centre National de la Recherche Scientifique ( CNRS ) -INSTITUT CURIE-Université Pierre et Marie Curie - Paris 6 ( UPMC ), Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Européen Georges Pompidou [APHP] ( HEGP ), Hôpital Beaujon [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Université Paris Diderot - Paris 7 (UPD7), and Centre National de la Recherche Scientifique (CNRS)-Institut Curie [Paris]-Université Pierre et Marie Curie - Paris 6 (UPMC)
Subjects: MESH : Molecular Weight, MESH : Immunohistochemistry, MESH : Immunoglobulin A, MESH : Models, Biological, MESH: Glomerulonephritis, IGA, digestive system, MESH : Gliadin, MESH : Chromatography, High Pressure Liquid, MESH: Biopsy, MESH: Enterocytes, MESH : Antigens, CD, [ SDV.IMM ] Life Sciences [q-bio]/Immunology, MESH : Receptors, Transferrin, MESH: Receptors, Transferrin, MESH: Immunoglobulin A, intestinal transport, MESH: Chromatography, High Pressure Liquid, MESH: Antigens, CD, Ussing chamber, MESH: Humans, MESH : Peptides, MESH: Peptides, MESH: Molecular Weight, MESH : Humans, MESH: Models, Biological, food and beverages, nutritional and metabolic diseases, MESH: Gliadin, MESH: Immunohistochemistry, MESH : Celiac Disease, transferrin receptor, digestive system diseases, MESH : Enterocytes, gliadin peptides, MESH : Biopsy, receptor-mediated transcytosis, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH : Glomerulonephritis, IGA, celiac disease, IgA, MESH: Celiac Disease
Abstract: International audience; Celiac disease (CD) is an enteropathy resulting from an abnormal immune response to gluten-derived peptides in genetically susceptible individuals. This immune response is initiated by intestinal transport of intact peptide 31-49 (p31-49) and 33-mer gliadin peptides through an unknown mechanism. We show that the transferrin receptor CD71 is responsible for apical to basal retrotranscytosis of gliadin peptides, a process during which p31-49 and 33-mer peptides are protected from degradation. In patients with active CD, CD71 is overexpressed in the intestinal epithelium and colocalizes with immunoglobulin (Ig) A. Intestinal transport of intact p31-49 and 33-mer peptides was blocked by polymeric and secretory IgA (SIgA) and by soluble CD71 receptors, pointing to a role of SIgA-gliadin complexes in this abnormal intestinal transport. This retrotranscytosis of SIgA-gliadin complexes may promote the entry of harmful gliadin peptides into the intestinal mucosa, thereby triggering an immune response and perpetuating intestinal inflammation. Our findings strongly implicate CD71 in the pathogenesis of CD.
Published: 2008

34. Clinical analysis of dendritic cell subsets: the dendritogram

Author: Hosmalin, Anne, Lichtner, Miriam, Louis, Stéphanie, Institut Cochin ( UMR_S567 / UMR 8104 ), 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 Cochin ( UM3 (UMR 8104 / U1016) ), J. Ewbank, E. Vivier, Institut Cochin (UMR_S567 / UMR 8104), 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 Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: MESH : Blood Cell Count, MESH: Dendritic Cells, MESH: Antibodies, MESH : Immunohistochemistry, MESH: Immunohistochemistry, hemic and immune systems, macromolecular substances, MESH: Biological Assay, MESH: Cell Separation, MESH : Antibodies, MESH : Leukocytes, Mononuclear, MESH: Leukocytes, Mononuclear, MESH : Dendritic Cells, MESH : Cell Separation, [SDV.IMM]Life Sciences [q-bio]/Immunology, [ SDV.IMM ] Life Sciences [q-bio]/Immunology, MESH: Animals, MESH : Animals, MESH : Biological Assay, MESH: Blood Cell Count
Abstract: International audience; Dendritic cells (DCs) are crucial in adaptive immunity because they are the only antigen-presenting cells that can present antigens to naive T lymphocytes. Plasmacytoid DCs (pDC) are also the main producers of type I Interferons in response to infection. We have shown that circulating myeloid DC (mDC) and pDC numbers are reduced in chronic as well as primary HIV infection. Data from different laboratories indicate that pDC counts, obtained by flow cytometry and rare event analysis, correlate inversely with the viral load, may be an early marker of recovery after antiretroviral treatment, and may predict better immune control of HIV replication. PDC counts may also be predictive of severe illness in dengue virus infection or of successful treatment against Mycobacterium tuberculosis. DC counts, or the "dendritogram", may therefore become useful in the clinical assessment of different infectious diseases.
Published: 2008

35. Incidence of amyloidosis over 3 years: the AMYPRO study

Author: Magy-Bertrand N, Jl, Dupond, Fred Mauny, As, Dupond, Duchene F, Gil H, Kantelip B, Laboratoire Chrono-environnement ( LCE ), Université Bourgogne Franche-Comté ( UBFC ) -Centre National de la Recherche Scientifique ( CNRS ) -Université de Franche-Comté ( UFC ), Laboratoire Chrono-environnement - CNRS - UBFC (UMR 6249) (LCE), Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), and Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)
Subjects: Male, Biopsy, MESH : Immunohistochemistry, MESH: Eyelids, MESH: Heart Valves, MESH : Aged, MESH : Prospective Studies, MESH : Carotid Arteries, MESH: Ligaments, Kidney, MESH: Biopsy, MESH: Aged, 80 and over, Prealbumin, MESH : Female, Prospective Studies, MESH: Incidence, Aged, 80 and over, MESH: Aged, MESH: Middle Aged, Incidence, Brain, [ SDV.SPEE ] Life Sciences [q-bio]/Santé publique et épidémiologie, Amyloidosis, Middle Aged, Heart Valves, Immunohistochemistry, MESH : Incidence, MESH : Kidney, MESH : Eyelids, Carotid Arteries, Adipose Tissue, MESH : Prealbumin, Female, France, MESH : Heart Valves, MESH: Adipose Tissue, Amyloid, MESH : Male, MESH : Adipose Tissue, MESH: Brain, MESH : Amyloid, MESH: Prealbumin, Humans, MESH: Amyloidosis, MESH : Middle Aged, MESH : France, MESH : Aged, 80 and over, Aged, MESH: Amyloid, Ligaments, MESH: Humans, MESH: Carotid Arteries, MESH : Humans, Eyelids, MESH: Immunohistochemistry, MESH: Kidney, MESH: Male, MESH: Prospective Studies, MESH : Amyloidosis, Gastrointestinal Tract, MESH: France, MESH : Brain, MESH : Biopsy, MESH: Gastrointestinal Tract, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, MESH : Gastrointestinal Tract, MESH : Ligaments, MESH: Female
Abstract: International audience; OBJECTIVES:There is a lack of epidemiological information concerning amyloidosis, particularly in France. We started a 3-year prospective study (AMPYRO) to analyze the epidemiological features of amyloidosis in the eastern part of France. METHODS:From 2003 to 2005, all patients with a tissue sample showing amyloid deposits, were included in this study. Immunohistochemistry using anti-P component, anti-SAA, anti-light chains immunoglobulins and anti-transthyretin was applied for each tissue sample. For each patient, past and present medical histories along with biological features were recorded. RESULTS:Seventy-six patients with amyloid were identified over 3 years. The age-standardized incidence rate of amyloidosis was estimated at 14 cases per million person-years. The final entire population included in the AMYPRO study was composed of 66 patients with a mean age of 71.7+/-11.5 years old. The amyloid typing after clinical, biological and immunohistochemistry revealed senile amyloid in 40 cases (60.6%), AL amyloid in 13 (19.7%) and AA amyloid in 9 (13.6%). Neither clinical nor biological features differed significantly between the transthyretin-positive and transthyretin-negative populations.CONCLUSION:Regarding only tissue samples, senile amyloid was the most prominent amyloid type identified. Therefore, the clinician needs to be aware that in most of the amyloid cases identified on the pathologic examination there is no need for additional examination unless there are clinical or biological signs of a primary or secondary amyloidosis.
Published: 2008

36. In vitro induction of differentiation by retinoic acid in an immortalized olfactory neuronal cell line

Author: Germaine Michel, Boris Lakard, Sophie Lakard, Claudine Versaux-Botteri, Eric Lesniewska, Nadege Morrand-Villeneuve, Laboratoire Interdisciplinaire Carnot de Bourgogne (ICB), Université de Bourgogne (UB)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Neurosciences Intégratives et Cliniques - UFC (UR 481) (NEURO), Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Laboratoire de Chimie des Matériaux et Interfaces (EA 474) (LCMI), Centre de Recherche des Cordeliers (CRC (UMR_S 872)), Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Ingénierie et biologie cellulaire et tissulaire (IBCT (ex IFR133)), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS BFC)-Université de Franche-Comté (UFC), Laboratoire Interdisciplinaire Carnot de Bourgogne ( LICB ), Centre National de la Recherche Scientifique ( CNRS ) -Université de Bourgogne ( UB ), Laboratoire de Neurosciences Intégratives et Cliniques - UFC ( NEURO ), Université Bourgogne Franche-Comté ( UBFC ) -Université de Franche-Comté ( UFC ), Laboratoire de Chimie des Matériaux et Interfaces ( LCMI ), Université de Franche-Comté ( UFC ), Centre de Recherche des Cordeliers ( CRC (UMR_S 872) ), Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Ingénierie et biologie cellulaire et tissulaire ( IBCT (ex IFR133) ), Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Etablissement français du sang [Bourgogne-France-Comté] ( EFS [Bourgogne-France-Comté] ) -Université de Franche-Comté ( UFC ), Laboratoire Interdisciplinaire Carnot de Bourgogne (LICB), Laboratoire de Neurosciences Intégratives et Cliniques - UFC (EA 481) (NEURO), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université de Franche-Comté (UFC), Laboratoire de Chimie des Matériaux et Interfaces (LCMI), Centre Hospitalier Régional Universitaire [Besançon] (CHRU Besançon)-Etablissement français du sang [Bourgogne-France-Comté] (EFS [Bourgogne-France-Comté])-Université de Franche-Comté (UFC), Université Paris Descartes - Paris 5 (UPD5)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Université de Franche-Comté (UFC), Université de Bourgogne ( UB ) -Centre National de la Recherche Scientifique ( CNRS ), Laboratoire de Neurosciences Intégratives et Cliniques - UFC (EA 481) ( NEURO ), and Université Bourgogne Franche-Comté [COMUE] ( UBFC ) -Université de Franche-Comté ( UFC )
Subjects: Time Factors, MESH : Immunohistochemistry, Retinoic acid, MESH: Neurons, MESH: Intermediate Filament Proteins, Apoptosis, MESH: Tubulin, Microscopy, Atomic Force, MESH : Adenylate Cyclase, Nestin, chemistry.chemical_compound, 0302 clinical medicine, Intermediate Filament Proteins, Tubulin, MESH : Cell Proliferation, MESH: Glial Fibrillary Acidic Protein, MESH: Animals, Oncogene E1A, MESH: Nerve Tissue Proteins, MESH : Nerve Tissue Proteins, MESH : Tretinoin, Cell Line, Transformed, Neurons, MESH: Microscopy, Atomic Force, 0303 health sciences, MESH : Rats, MESH : Cell Line, Transformed, Cell Differentiation, General Medicine, Transfection, MESH : Tubulin, Immunohistochemistry, MESH : Glial Fibrillary Acidic Protein, Cell biology, medicine.anatomical_structure, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], MESH : Cell Differentiation, Adenylyl Cyclases, MESH : Time Factors, MESH : Microscopy, Electron, Scanning, MESH: Cell Differentiation, Histology, MESH: Microscopy, Electron, Scanning, MESH: Rats, Immunocytochemistry, Nerve Tissue Proteins, Tretinoin, Biology, MESH : Neurons, 03 medical and health sciences, Olfactory Mucosa, MESH: Cell Proliferation, Glial Fibrillary Acidic Protein, MESH: Adenylate Cyclase, medicine, Animals, MESH: Cell Line, Transformed, Progenitor cell, MESH: Olfactory Mucosa, Cell Proliferation, 030304 developmental biology, MESH : Olfactory Mucosa, MESH: Tretinoin, MESH: Apoptosis, MESH: Time Factors, MESH: Immunohistochemistry, Cell Biology, Molecular biology, MESH : Microscopy, Atomic Force, Rats, chemistry, Cell culture, [ SDV.NEU ] Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Microscopy, Electron, Scanning, MESH : Animals, MESH : Intermediate Filament Proteins, Olfactory epithelium, 030217 neurology & neurosurgery, MESH : Apoptosis
Abstract: International audience; In this study, we used a neuronal cell line generated by transfection of rat olfactory epithelium with immortalizing recombinant oncogene E1A of adenovirus-2. The resulting 13.S.1.24 line of transformed cells expressed an antigenic phenotype of olfactory neuronal progenitors. Time-dependency assessments over 1 week of treatment indicated that apoptosis and differentiation induced by retinoic acid (RA) were concomitant. Indeed, RA altered the cell proliferation rate, but it also stimulated differentiation of surviving 13.S.1.24 cells into bipolar olfactory marker protein-immunoreactive neurons. To characterize the nature of the cells we used immunocytochemistry, optical imaging, scanning electron microscopy and atomic force microscopy.
Published: 2007

37. Protein mapping of calcium carbonate biominerals by immunogold

Author: Boaz Pokroy, Gilles Luquet, Klaas de Groot, Pierre Layrolle, Frédéric Marin, Faculty of Science and Technology, Biomaterials Science and Technology, Layrolle, Pierre, Biogéosciences [UMR 6282] (BGS), Université de Bourgogne (UB)-Centre National de la Recherche Scientifique (CNRS), Department of Materials Engineering, Technion - Israel Institute of Technology [Haifa], Laboratoire d'ingénierie osteo-articulaire et dentaire (LIOAD), Université de Nantes (UN)-IFR26-Institut National de la Santé et de la Recherche Médicale (INSERM), Institute for Biomedical Technology, University of Twente, Biogéosciences [Dijon] ( BGS ), Université de Bourgogne ( UB ) -AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique ( CNRS ), Laboratoire d'ingénierie osteo-articulaire et dentaire ( LIOAD ), Université de Nantes ( UN ) -IFR26-Institut National de la Santé et de la Recherche Médicale ( INSERM ), University of Twente [Netherlands], Biogéosciences [UMR 6282] [Dijon] (BGS), and Université de Bourgogne (UB)-AgroSup Dijon - Institut National Supérieur des Sciences Agronomiques, de l'Alimentation et de l'Environnement-Centre National de la Recherche Scientifique (CNRS)
Subjects: MESH : Models, Chemical, MESH : Molecular Sequence Data, MESH: Sequence Homology, Amino Acid, MESH : Calcium Carbonate, MESH : Immunohistochemistry, MESH : Aspartic Acid, MESH: Trypsin, MESH: Amino Acid Sequence, Matrix (biology), 01 natural sciences, MESH: Aspartic Acid, MESH : Proteins, chemistry.chemical_compound, Trypsin, MESH: Animals, MESH: Proteins, Peptide sequence, MESH: Crystallization, chemistry.chemical_classification, 0303 health sciences, Caspartin, biology, MESH : Amino Acid Sequence, MESH : Pepsin A, MESH: Models, Chemical, Immunogold labelling, Immunohistochemistry, MESH: Mollusca, MESH : Sequence Homology, Amino Acid, Amino acid, Biochemistry, MESH: Calcium Carbonate, Mechanics of Materials, MESH : Crystallization, MESH: Pepsin A, SEM, MESH : Edetic Acid, Crystallization, MESH : Mollusca, Calcium carbonate, Proteinaceous extracellular matrix, MESH: Edetic Acid, Molecular Sequence Data, Biophysics, Bioengineering, 010402 general chemistry, Biomaterials, 03 medical and health sciences, Animals, Amino Acid Sequence, [SDV.IB.BIO]Life Sciences [q-bio]/Bioengineering/Biomaterials, Edetic Acid, 030304 developmental biology, Aspartic Acid, Viral matrix protein, MESH: Molecular Sequence Data, Sequence Homology, Amino Acid, MESH : Solubility, Back-scattered electrons, Surface treatmen, Proteins, MESH: Immunohistochemistry, IR-78873, [ SDV.IB.BIO ] Life Sciences [q-bio]/Bioengineering/Biomaterials, Pepsin A, 0104 chemical sciences, [SDV.IB.BIO] Life Sciences [q-bio]/Bioengineering/Biomaterials, MESH: Solubility, chemistry, Models, Chemical, Solubility, Polyclonal antibodies, Mollusca, Ceramics and Composites, biology.protein, MESH : Animals, MESH : Trypsin, Immunogold
Abstract: The construction of metazoan calcium carbonate skeletons is finely regulated by a proteinaceous extracellular matrix, which remains embedded within the exoskeleton. In spite of numerous biochemical studies, the precise localization of skeletal proteins has remained for a long time as an elusive goal. In this paper, we describe a technique for visualizing shell matrix proteins on the surface of calcium carbonate crystals or within the biominerals. The technique is as follows: freshly broken pieces of biominerals or NaOCl then EDTA-etched polished surfaces are incubated with an antibody elicited against one matrix protein, then with a secondary gold-coupled antibody. After silver enhancement, the samples are subsequently observed with scanning electron microscopy by using back-scattered electron mode. In the present case, the technique is applied to a particular example, the calcitic prisms that compose the outer shell layer of the mediterranean fan mussel Pinna nobilis. One major soluble protein, caspartin, which was identified recently, was partly de novo sequenced after enzymatic digestions. A polyclonal antibody raised against caspartin was used for its localization within and on the prisms. The immunogold localization indicated that caspartin surrounds the calcitic prisms, but is also dispersed within the biominerals. This example illustrates the deep impact of the technique on the definition of intracrystalline versus intercrystalline matrix proteins. Furthermore, it is an important tool for assigning a putative function to a matrix protein of interest.
Published: 2007

38. Stromal cell networks regulate lymphocyte entry, migration, and territoriality in lymph nodes

Author: Ronald N. Germain, Frédéric Brau, Nicolas Glaichenhaus, Jackson G. Egen, Lily Koo, Marc Bajénoff, Jean Pierre Laugier, Lymphocyte Biology Section ( NIAID ), National Institutes of Health, Immunologie des Maladies Infectieuses Allergiques et Autoimmunes, Université Nice Sophia Antipolis ( UNS ), Université Côte d'Azur ( UCA ) -Université Côte d'Azur ( UCA ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Centre Commun de Microscopie Appliquée ( CCMA ), Université Côte d'Azur ( UCA ) -Université Côte d'Azur ( UCA ), Institut de pharmacologie moléculaire et cellulaire ( IPMC ), Université Côte d'Azur ( UCA ) -Université Côte d'Azur ( UCA ) -Centre National de la Recherche Scientifique ( CNRS ), Intramural Research Program of NIAID, NIH, DHHS, Institut de la Santé et de la Recherche Médicale (INSERM), Lymphocyte Biology Section (NIAID), Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre Commun de Microscopie Appliquée (CCMA), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA), Institut de pharmacologie moléculaire et cellulaire (IPMC), and 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)
Subjects: Pathology, Adoptive cell transfer, Lymphocyte, MESH : Immunohistochemistry, MESH : Diagnostic Imaging, MESH : Lymphocytes, MESH: Chemotaxis, Leukocyte, MESH: Lymph Nodes, Cell Communication, Mice, 0302 clinical medicine, Reticular cell, MESH: Microscopy, Confocal, Immunology and Allergy, [ SDV.IMM ] Life Sciences [q-bio]/Immunology, MESH: Animals, Lymphocytes, MESH : Lymph Nodes, 0303 health sciences, Microscopy, Confocal, MESH : Adoptive Transfer, Cell migration, MESH : Chemotaxis, Leukocyte, Adoptive Transfer, Immunohistochemistry, Cell biology, Chemotaxis, Leukocyte, medicine.anatomical_structure, Infectious Diseases, MESH : Microscopy, Electron, Transmission, MESH: Microscopy, Electron, Transmission, [SDV.IMM]Life Sciences [q-bio]/Immunology, Diagnostic Imaging, MESH : Microscopy, Electron, Scanning, medicine.medical_specialty, Stromal cell, Naive T cell, MESH: Microscopy, Electron, Scanning, MESH : Cell Communication, T cell, Immunology, MESH : Mice, Inbred C57BL, Biology, 03 medical and health sciences, Microscopy, Electron, Transmission, MESH: Mice, Inbred C57BL, MESH: Cell Communication, MESH : Mice, medicine, Animals, MESH : Microscopy, Confocal, MESH: Mice, 030304 developmental biology, Follicular dendritic cells, MESH: Diagnostic Imaging, MESH: Immunohistochemistry, Mice, Inbred C57BL, MESH: Adoptive Transfer, CELLIMMUNO, MESH : Stromal Cells, Microscopy, Electron, Scanning, CELLBIO, MESH: Lymphocytes, Lymph Nodes, MESH : Animals, Stromal Cells, MESH: Stromal Cells, 030215 immunology
Abstract: SummaryAfter entry into lymph nodes (LNs), B cells migrate to follicles, whereas T cells remain in the paracortex, with each lymphocyte type showing apparently random migration within these distinct areas. Other than chemokines, the factors contributing to this spatial segregation and to the observed patterns of lymphocyte movement are poorly characterized. By combining confocal, electron, and intravital microscopy, we showed that the fibroblastic reticular cell network regulated naive T cell access to the paracortex and also supported and defined the limits of T cell movement within this domain, whereas a distinct follicular dendritic cell network similarly served as the substratum for movement of follicular B cells. These results highlight the central role of stromal microanatomy in orchestrating cell migration within the LN.
Published: 2006

39. Partial defects in transcriptional activity of two novel DAX-1 mutations in childhood-onset adrenal hypoplasia congenita

Author: Paul Laissue, Silvia Copelli, Marc Fellous, Gabriel Barrio, César Bergadá, Sinan Karaboga, Jean Marie Wurtz, Reiner A. Veitia, Ignacio Bergadá, Enzo Lalli, Institut Cochin (UMR_S567 / UMR 8104), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Departamento de Ciencias Biologicas, Universidad CAECE, Centro de Investigaciones Endocrinológicas (CEDIE), Hospital de Niños 'Ricardo Gutiérrez', 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), Institut de pharmacologie moléculaire et cellulaire (IPMC), Université Nice Sophia Antipolis (... - 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), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Descartes - Paris 5 (UPD5), Université Côte d'Azur (UCA)-Université Côte d'Azur (UCA)-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), Université Louis Pasteur - Strasbourg I - Institut National de la Santé et de la Recherche Médicale (INSERM) - Centre National de la Recherche Scientifique (CNRS), Université Nice Sophia Antipolis (UNS), and Université Côte d'Azur (UCA) - Université Côte d'Azur (UCA) - Centre National de la Recherche Scientifique (CNRS)
Subjects: Male, Transcription, Genetic, MESH : Hela Cells, Receptors, Retinoic Acid, Endocrinology, Diabetes and Metabolism, MESH : Immunohistochemistry, DNA Mutational Analysis, Mutant, MESH : Child, Preschool, MESH: Protein Structure, Tertiary, 0302 clinical medicine, Endocrinology, MESH: Structure-Activity Relationship, MESH : Child, MESH: Child, Adrenal Glands, MESH : Structure-Activity Relationship, MESH: Adrenal Glands, MESH: DNA Mutational Analysis, Child, 0303 health sciences, [SDV.MHEP] Life Sciences [q-bio]/Human health and pathology, MESH : Infant, Transfection, Immunohistochemistry, Protein subcellular localization prediction, MESH: Infant, MESH : Receptors, Retinoic Acid, DNA-Binding Proteins, MESH: Repressor Proteins, Child, Preschool, MESH: Adrenal Insufficiency, MESH : DNA-Binding Proteins, MESH : Mutation, MESH : Repressor Proteins, MESH : Transfection, MESH : Protein Structure, Tertiary, medicine.medical_specialty, MESH: Mutation, MESH : Male, 030209 endocrinology & metabolism, [SDV.CAN]Life Sciences [q-bio]/Cancer, MESH : DNA Mutational Analysis, Biology, MESH : Adrenal Glands, Structure-Activity Relationship, 03 medical and health sciences, [SDV.CAN] Life Sciences [q-bio]/Cancer, Internal medicine, X-linked adrenal hypoplasia congenita, medicine, [SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, Epigenetics, Gene, 030304 developmental biology, MESH: Receptors, Retinoic Acid, MESH: Humans, DAX-1 Orphan Nuclear Receptor, MESH: Transcription, Genetic, MESH: Transfection, MESH : Humans, MESH: Child, Preschool, Infant, MESH : Transcription, Genetic, MESH: Immunohistochemistry, MESH : Adrenal Insufficiency, medicine.disease, MESH: Male, Protein Structure, Tertiary, Repressor Proteins, MESH: Hela Cells, Nuclear receptor, Mutation, DAX1, MESH: DNA-Binding Proteins, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology, Adrenal Insufficiency, HeLa Cells
Abstract: OBJECTIVE: Mutations in DAX-1, an X-linked gene encoding an orphan nuclear receptor, have been associated with adrenal hypoplasia congenita and hypogonadotropic hypogonadism. Here we describe two novel DAX-1 mutations, Y214X and I361T, associated with childhood-onset primary adrenal failure. We aimed at analysing their effects on protein localization, transcriptional activity and propose a structural-function relationship. DESIGN: We have directly sequenced the DAX-1 gene from PCR-amplified DNA. The effect of the mutations on protein localization was assessed by immunocytochemistry in transfected cells. The impact of mutations on transcriptional activity was determined by transfection using a StAR promoter-luciferase reporter system. RESULTS: The mutation Y214X produces a protein lacking the C-terminal half of DAX-1. The other one, I361T, affects a highly conserved amino acid within the putative ligand-binding domain. The mutant Y214X displayed a wild-type subcellular localization while I361T was partially retained in the cytoplasm. Both mutants retained subtantial transcriptional repressive activity, compared to mutants producing early onset adrenal failure. Interestingly, I361T displayed similar in vitro transcriptional repressive activity to the mutant I439S previously described in a case of late-onset AHC. This shows that molecular alterations of DAX-1 leading to similar in vitro activities may result in very different ages of onset of adrenal failure, which suggests that additional genetic and epigenetic factors are important in determining the clinical course of AHC. CONCLUSIONS: Our results help the understanding of structure-function relationships in the DAX-1 molecule, suggesting that the N-terminal domain is relatively autonomous and add credence to presumed molecular interactions within ligand binding domain of the protein.
Published: 2006

40. Expression and localization of alpha-fetoprotein mRNA and protein in human early villous trophoblasts

Author: F. Ferré, A.M. Hagneré, Daniel Vaiman, Françoise Mondon, Thérèse-Marie Mignot, C. Benassayag, B. Robert, P. Duc-Goiran, Marie-Cécile Vacher-Lavenu, J.L. Danan, F. Machavoine, Génomique et épigénétique des pathologies placentaires (Inserm U709), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Descartes - Paris 5 (UPD5), Cytokines, hématopoïèse et réponse immune (CHRI), 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 d'Anatomie et de cytologie pathologiques [CHU Cochin], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Biologie des Transporteurs Mitochondriaux et Métabolisme (BIOTRAM), Centre National de la Recherche Scientifique (CNRS), ProdInra, Migration, Université Paris Descartes - Paris 5 (UPD5) - Institut National de la Santé et de la Recherche Médicale (INSERM) - Centre National de la Recherche Scientifique (CNRS), Génomique et épigénétique des pathologies placentaires ( Inserm U709 ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Paris Descartes - Paris 5 ( UPD5 ), Cytokines, hématopoïèse et réponse immune ( CHRI ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Centre National de la Recherche Scientifique ( CNRS ), Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Cochin [AP-HP], Biologie des Transporteurs Mitochondriaux et Métabolisme ( BIOTRAM ), Centre National de la Recherche Scientifique ( CNRS ), Slama, Catherine, Unité de recherche Génétique Biochimique et Cytogénétique (LGBC), Institut National de la Recherche Agronomique (INRA), and Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Cochin [AP-HP]
Subjects: MESH : RNA, Messenger, MESH: Trophoblasts, [SDV]Life Sciences [q-bio], MESH : Immunohistochemistry, ALPHA-FETOPROTEINE, Gene Expression, MESH : Chorionic Villi, 0302 clinical medicine, MESH: Pregnancy, Pregnancy, Gene expression, MESH : Female, PLACENTA, MESH: Chorionic Villi, ComputingMilieux_MISCELLANEOUS, In Situ Hybridization, 0303 health sciences, digestive, oral, and skin physiology, MESH : In Situ Hybridization, Obstetrics and Gynecology, Immunohistochemistry, Trophoblasts, [SDV] Life Sciences [q-bio], medicine.anatomical_structure, 030220 oncology & carcinogenesis, embryonic structures, Female, MESH : Trophoblasts, alpha-Fetoproteins, Chorionic Villi, Alpha-fetoprotein, TROPHOBLATE, MESH: Gene Expression, ALPHA-FETOPROTEIN, In situ hybridization, Biology, TROPHOBAST, 03 medical and health sciences, MESH: In Situ Hybridization, Fetal membrane, Placenta, medicine, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, RNA, Messenger, Yolk sac, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, 030304 developmental biology, MESH: RNA, Messenger, Fetus, MESH: Humans, MESH : Humans, MESH: Immunohistochemistry, Molecular biology, digestive system diseases, MESH : Gene Expression, MESH : Pregnancy, MESH : alpha-Fetoproteins, Reproductive Medicine, MESH: alpha-Fetoproteins, MESH: Female, Developmental Biology
Abstract: Alpha-fetoprotein (AFP) is a major plasma protein produced during human fetal life. It is a good marker for several possible disorders affecting gestation. We previously reported that afp gene expression, which takes place mainly in yolk sac and fetal liver, also occurs in normal human placenta, specifically in early pregnancy. The aim of the present study was to determine the precise location of AFP synthesis sites within the placental villi. In situ hybridization and immunohistochemical experiments were performed on sections obtained from placentas of first-trimester and full-term pregnancies. We found that the pattern of afp gene expression was restricted to specific villous trophoblastic areas in early placentas. Both afp transcripts and AFP protein were mainly located in discontinuous regions, at junctions between two villi and at budding sites. In contrast, no AFP expression was detected in the cytotrophoblastic extravillous proliferative zone or in other placental cell types. According to the earlier studies, no AFP synthesis was detected in placental villous tissue from full-term pregnancies, using in situ hybridization and immunohistochemistry.
Published: 2006

41. Immunohistochemical analysis of CD4+ and CD8+ T-cell subsets in high risk human papillomavirus-associated pre-malignant and malignant lesions of the uterine cervix

Author: Christiane Mougin, Frédéric Mauny, Sylvain Monnier-Benoit, Estelle Seilles, Mihai Căpîlna, Jean-Luc Prétet, Sophie Félix, Didier Riethmuller, Jean-Sébastien Guerrini, Carcinogénèse épithéliale : facteurs prédictifs et pronostiques - UFC ( CEF2P / CARCINO ), Centre Hospitalier Régional Universitaire [Besançon] ( CHRU Besançon ) -Université Bourgogne Franche-Comté ( UBFC ) -Université de Franche-Comté ( UFC ), Département d'information médicale, Hôpital Saint-Jacques, Clinica de Obstetrică-Ginecologie, Târgu Mureş Hospital, Service d'Anatomopathologie, hopital Jean Minjoz, Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC ( HOTE GREFFON ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Etablissement français du sang [Bourgogne-France-Comté] ( EFS [Bourgogne-France-Comté] ) -Université de Franche-Comté ( UFC ), Carcinogénèse épithéliale : facteurs prédictifs et pronostiques - UFC (EA 3181) (CEF2P / CARCINO), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC), Centre Hospitalier Régional Universitaire de Besançon (CHRU Besançon), Interactions hôte-greffon-tumeur, ingénierie cellulaire et génique - UFC (UMR INSERM 1098) (RIGHT), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement français du sang [Bourgogne-Franche-Comté] (EFS [Bourgogne-Franche-Comté])-Université de Franche-Comté (UFC)
Subjects: CD4-Positive T-Lymphocytes, Pathology, MESH : Immunohistochemistry, Uterine Cervical Neoplasms, MESH: T-Lymphocyte Subsets, CD8-Positive T-Lymphocytes, MESH : Cervical Intraepithelial Neoplasia, MESH: Papillomavirus Infections, MESH : Neoplasm Invasiveness, 0302 clinical medicine, MESH: Cell Aggregation, MESH: Papillomaviridae, T-Lymphocyte Subsets, MESH : Antigens, CD45, Cytotoxic T cell, MESH : Female, Papillomaviridae, Cell Aggregation, MESH : Papillomavirus Infections, 0303 health sciences, biology, Obstetrics and Gynecology, MESH: CD4-Positive T-Lymphocytes, [ SDV.SPEE ] Life Sciences [q-bio]/Santé publique et épidémiologie, MESH : CD8-Positive T-Lymphocytes, Immunohistochemistry, MESH: CD8-Positive T-Lymphocytes, female genital diseases and pregnancy complications, 3. Good health, MESH: Uterine Cervical Neoplasms, Oncology, 030220 oncology & carcinogenesis, MESH : CD4-Positive T-Lymphocytes, Disease Progression, Female, MESH: Disease Progression, medicine.medical_specialty, medicine.drug_class, MESH : Uterine Cervical Neoplasms, MESH: Antigens, CD45, Monoclonal antibody, 03 medical and health sciences, Immune system, Stroma, medicine, Humans, Neoplasm Invasiveness, MESH : Papillomaviridae, MESH : Cell Aggregation, 030304 developmental biology, MESH: Humans, business.industry, Papillomavirus Infections, MESH : Humans, Cancer, MESH: Immunohistochemistry, MESH: Neoplasm Invasiveness, MESH : Disease Progression, Uterine Cervical Dysplasia, biology.organism_classification, medicine.disease, MESH : T-Lymphocyte Subsets, Leukocyte Common Antigens, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, business, MESH: Cervical Intraepithelial Neoplasia, MESH: Female, CD8
Abstract: International audience; OBJECTIVES: Humoral and cellular immune responses are likely to play a key role for the clearance or persistence and progression of high risk (HR) HPV-associated cervical lesions. Although there are many studies describing the systemic T-cell responses to HPV16 and 18 proteins, few data are available regarding the cellular mucosal immune responses. We used immunohistochemistry to characterize populations of T-immune cells (CD4+, CD8+, CD45RO+) in HR-HPV-infected precancerous and cancerous lesions of the uterine cervix. METHODS: Four biopsies from normal cervix, 9 CIN1 which have regressed (rCIN), 5 CIN1 which have progressed (pCIN) to high grade lesions, 13 CIN3 and 11 invasive carcinomas were included. All dysplasias and carcinomas were HR-HPV-positive and low-risk-HPV-negative. They were stained with monoclonal antibodies specific for CD4, CD8 and CD45RO and examined by microscopy. STATISTICAL ANALYSIS: The Kruskal-Wallis test and the Siegel's and Castelan's method were used. RESULTS.: CD4+ cells predominated in regressing CIN1 both within the stroma and the epithelium with the highest CD4+/CD8+ ratio compared with pCIN1, CIN3 and invasive carcinoma. At the exception of CD45RO+ cells, T cells were detected with similar frequencies in both pCIN1 and CIN3. However, in 7 out of 10 CIN3, CD4+ and CD8+ cells were visible as organized lymphoid follicle structure. The CD8+ and CD45RO+ cells far exceeded the CD4+ cells in invasive cancers. CONCLUSIONS: Density and distribution of immune T cells depend on the malignant potential of HR-HPV lesions. These results suggest that the studied lymphocyte subsets have an important role to fulfil during the natural history of HR-HPV-associated lesions.
Published: 2006

42. Natural killer cell behavior in lymph nodes revealed by static and real-time imaging

Author: Béatrice Breart, Hai Qi, Veronique M. Braud, Marc Bajénoff, Ronald N. Germain, Alex Yee-Chen Huang, Nicolas Glaichenhaus, Julie Cazareth, Institut de pharmacologie moléculaire et cellulaire ( IPMC ), Université Nice Sophia Antipolis ( UNS ), Université Côte d'Azur ( UCA ) -Université Côte d'Azur ( UCA ) -Centre National de la Recherche Scientifique ( CNRS ), National Institutes of Health, Bethesda ( NIH ), National Institutes of Health ( NIH ), Université Côte d'Azur ( UCA ), Immunologie des Maladies Infectieuses Allergiques et Autoimmunes, Université Côte d'Azur ( UCA ) -Université Côte d'Azur ( UCA ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Institut de pharmacologie moléculaire et cellulaire (IPMC), Université Nice Sophia Antipolis (... - 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), National Institutes of Health, Bethesda (NIH), National Institutes of Health [Bethesda] (NIH), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA), and COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Institut National de la Santé et de la Recherche Médicale (INSERM)
Subjects: CD4-Positive T-Lymphocytes, MESH: Signal Transduction, MESH: Inflammation, Cellular differentiation, MESH : Immunohistochemistry, MESH: Lymph Nodes, MESH : Leishmaniasis, Cutaneous, Lymphocyte Activation, Mice, Interleukin 21, 0302 clinical medicine, MESH: Leishmania major, Immunology and Allergy, Interferon gamma, MESH: Animals, MESH : Lymph Nodes, Leishmania major, Mice, Inbred BALB C, 0303 health sciences, MESH: CD4-Positive T-Lymphocytes, Cell Differentiation, MESH : Interferon Type II, Articles, Acquired immune system, Immunohistochemistry, MESH : Mice, Transgenic, Cell biology, Killer Cells, Natural, medicine.anatomical_structure, MESH : CD4-Positive T-Lymphocytes, MESH : Killer Cells, Natural, MESH : Cell Differentiation, Intravital microscopy, Signal Transduction, medicine.drug, MESH: Cell Differentiation, MESH: Killer Cells, Natural, MESH: Mice, Transgenic, MESH: Interferon Type II, T cell, Immunology, MESH: Mice, Inbred BALB C, Leishmaniasis, Cutaneous, Mice, Transgenic, Biology, Article, Natural killer cell, MESH : Leishmania major, Interferon-gamma, 03 medical and health sciences, MESH : Microscopy, Fluorescence, Multiphoton, MESH : Mice, medicine, Animals, Humans, [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Lymphocyte Activation, MESH: Mice, [ SDV.BBM ] Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH : Lymphocyte Activation, MESH : Mice, Inbred BALB C, 030304 developmental biology, Inflammation, MESH : Signal Transduction, MESH: Humans, MESH : Inflammation, MESH : Humans, MESH: Immunohistochemistry, Dendritic cell, Cell Biology, MESH: Leishmaniasis, Cutaneous, Microscopy, Fluorescence, Multiphoton, MESH: Microscopy, Fluorescence, Multiphoton, Lymph Nodes, MESH : Animals, 030215 immunology
Abstract: Natural killer (NK) cells promote dendritic cell (DC) maturation and influence T cell differentiation in vitro. To better understand the nature of the putative interactions among these cells in vivo during the early phases of an adaptive immune response, we have used immunohistochemical analysis and dynamic intravital imaging to study NK cell localization and behavior in lymph nodes (LNs) in the steady state and shortly after infection with Leishmania major. In the LNs of naive mice, NK cells reside in the medulla and the paracortex, where they closely associate with DCs. In contrast to T cells, intravital microscopy revealed that NK cells in the superficial regions of LNs were slowly motile and maintained their interactions with DCs over extended times in the presence or absence of immune-activating signals. L. major induced NK cells to secrete interferon-γ and to be recruited to the paracortex, where concomitant CD4 T cell activation occurred. Therefore, NK cells form a reactive but low mobile network in a strategic area of the LN where they can receive inflammatory signals, interact with DCs, and regulate colocalized T cell responses.
Published: 2006

43. Hepatocellular adenoma displaying a HNF1alpha inactivation in a patient with familial adenomatous polyposis coli

Author: François Paye, Najat Mourra, Dominique Wendum, Jean-François Fléjou, Claudia de Toma, Emmanuelle Jeannot, Jessica Zucman-Rossi, Zucman-Rossi, Jessica, Génomique Fonctionnelle des Tumeurs Solides (U1162), Université Paris 13 (UP13)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service d'anatomie et cytologie pathologiques [CHU Saint-Antoine], Université Pierre et Marie Curie - Paris 6 (UPMC)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Saint-Antoine [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), Service de chirurgie générale et digestive [CHU Saint-Antoine], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Saint-Antoine [AP-HP], Centre d'Etude du Polymorphisme Humain (CEPH), Université Paris Diderot - Paris 7 (UPD7)-Institut Universitaire d'Hématologie (IUH), Université Paris Diderot - Paris 7 (UPD7)-Fondation Jean Dausset, Genomique Fonctionnelle des Tumeurs Solides, Université Paris Diderot - Paris 7 ( UPD7 ) -IFR105-Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Service d'anatomie et cytologie pathologiques, Université Pierre et Marie Curie - Paris 6 ( UPMC ) -Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Saint-Antoine [APHP], Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Saint-Antoine [APHP], Centre d'Etude du Polymorphisme Humain ( CEPH ), Université Paris Diderot - Paris 7 ( UPD7 ) -Institut Universitaire d'Hématologie ( IUH ), and Université Paris Diderot - Paris 7 ( UPD7 ) -Fondation Jean Dausset
Subjects: Biallelic Mutation, MESH : Germ-Line Mutation, MESH : Immunohistochemistry, [SDV.GEN] Life Sciences [q-bio]/Genetics, MESH : Liv, Gene mutation, medicine.disease_cause, [ SDV.CAN ] Life Sciences [q-bio]/Cancer, 0302 clinical medicine, MESH: Germ-Line Mutation, MESH : Female, MESH: Gene Silencing, Hepatocyte Nuclear Factor 1-alpha, MESH: Carcinoma, Hepatocellular, MESH: Liv, MESH: Genes, APC, biology, Reverse Transcriptase Polymerase Chain Reaction, Liver Neoplasms, Wnt signaling pathway, MESH: Genetic Predisposition to Disease, MESH : Adult, MESH : Diagnosis, Differential, Immunohistochemistry, 3. Good health, Adenomatous Polyposis Coli, 030220 oncology & carcinogenesis, Female, MESH : Hepatocyte Nuclear Factor 1-alpha, [ SDV.MHEP.HEG ] Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, 030211 gastroenterology & hepatology, Adult, MESH : Carcinoma, Hepatocellular, congenital, hereditary, and neonatal diseases and abnormalities, Carcinoma, Hepatocellular, Genes, APC, Adenoma, Adenomatous polyposis coli, [SDV.CAN]Life Sciences [q-bio]/Cancer, Article, Diagnosis, Differential, 03 medical and health sciences, Germline mutation, [SDV.CAN] Life Sciences [q-bio]/Cancer, MESH: Diagnosis, Differential, MESH : Gene Silencing, medicine, Humans, Genetic Predisposition to Disease, Gene Silencing, MESH: Hepatocyte Nuclear Factor 1-alpha, Alleles, Germ-Line Mutation, [SDV.GEN]Life Sciences [q-bio]/Genetics, MESH: Humans, MESH : Adenomatous Polyposis Coli, Hepatology, MESH: Alleles, MESH : Humans, MESH: Adult, MESH: Immunohistochemistry, [SDV.MHEP.HEG]Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, Hepatocellular adenoma, medicine.disease, MESH: Adenomatous Polyposis Coli, digestive system diseases, [SDV.MHEP.HEG] Life Sciences [q-bio]/Human health and pathology/Hépatology and Gastroenterology, MESH : Genes, APC, biology.protein, Cancer research, MESH : Genetic Predisposition to Disease, MESH : Alleles, [ SDV.GEN ] Life Sciences [q-bio]/Genetics, Carcinogenesis, MESH: Female
Abstract: Patients with familial adenomatous polyposis coli (FAP) may rarely develop hepatocellular adenoma. Here we report the case of a 37-year-old FAP woman presenting a hepatocellular adenoma after oestroprogestative oral contraception use. In this steatotic adenoma, we identified an inactivating biallelic mutation of HNF1alpha. In addition to the known germline APC mutation Q1062fs, we did not find an inactivation of the second APC allele nor an activation of the beta-catenin target genes GLUL and GPR49. Our findings contrast with two hepatocellular adenoma cases related to FAP, for which a biallelic inactivation of the APC gene was previously described. Altogether, these results suggest that benign hepatocellular carcinogenesis may be dependent on or independent of the Wnt/beta-catenin pathway in patients with FAP.
Published: 2006

44. Angiogenin distribution in human term placenta, and expression by cultured trophoblastic cells

Author: Josette Badet, Danièle Evain-Brion, Yann Bassaglia, Jean-Louis Frendo, Elissavet Hatzi, Nadine Pavlov, Développement humain : Croissance et différenciation, 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), Laboratory of Biological Chemistry, University of Ioannina, Laboratoire de recherche sur la croissance cellulaire, la réparation et la régénération tissulaires (LRCCRRT), Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Centre National de la Recherche Scientifique (CNRS), INSERM, Ministère de la Recherche et de la Technologie, Association de la Recherche sur le Cancer, Ligue contre le Cancer, Novo Nordisk Pharmaceutique S.A., 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 Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), 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), Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12) - Centre National de la Recherche Scientifique (CNRS), and Badet, Josette
Subjects: Cancer Research, Physiology, Angiogenesis, MESH: Trophoblasts, MESH : Immunohistochemistry, Placenta, Clinical Biochemistry, MESH: Ribonuclease, Pancreatic, MESH : Ribonuclease, Pancreatic, Gene Expression, MESH : Pregnancy Trimester, Third, MESH : Tissue Distribution, MESH : Fluorescent Antibody Technique, Indirect, chemistry.chemical_compound, 0302 clinical medicine, MESH: Pregnancy, Pregnancy, MESH: Reverse Transcriptase Polymerase Chain Reaction, MESH : Female, Tissue Distribution, Fluorescent Antibody Technique, Indirect, MESH : Culture Media, Conditioned, Tissue homeostasis, reproductive and urinary physiology, Cells, Cultured, In Situ Hybridization, 0303 health sciences, MESH : Neovascularization, Physiologic, MESH: Culture Media, Conditioned, Reverse Transcriptase Polymerase Chain Reaction, MESH : Reverse Transcriptase Polymerase Chain Reaction, Trophoblast, MESH : In Situ Hybridization, MESH: Enzyme-Linked Immunosorbent Assay, MESH: Placenta, MESH : Enzyme-Linked Immunosorbent Assay, Immunohistochemistry, Trophoblasts, Endothelial stem cell, Vascular endothelial growth factor, medicine.anatomical_structure, MESH : Placenta, Placental ribonuclease inhibitor, 030220 oncology & carcinogenesis, embryonic structures, MESH: Pregnancy Trimester, Third, Female, MESH : Trophoblasts, MESH: Neovascularization, Physiologic, MESH: Cells, Cultured, MESH: Gene Expression, Angiogenin, Pregnancy Trimester, Third, Neovascularization, Physiologic, Enzyme-Linked Immunosorbent Assay, Biology, Article, 03 medical and health sciences, Syncytiotrophoblast, MESH: In Situ Hybridization, MESH : Cells, Cultured, medicine, MESH: Fluorescent Antibody Technique, Indirect, Humans, MESH: Tissue Distribution, [SDV.BDLR] Life Sciences [q-bio]/Reproductive Biology, 030304 developmental biology, MESH: Humans, MESH : Humans, MESH: Biological Markers, MESH: Immunohistochemistry, [SDV.BDLR]Life Sciences [q-bio]/Reproductive Biology, Ribonuclease, Pancreatic, Molecular biology, MESH : Biological Markers, MESH : Gene Expression, MESH : Pregnancy, chemistry, Culture Media, Conditioned, MESH: Female, Biomarkers
Abstract: International audience; Human angiogenin is a 14-kDa secreted protein with angiogenic and ribonucleolytic activities. Angiogenin is associated with tumour development but is also present in normal biological fluids and tissues. To further address the physiological role of angiogenin, we studied its expression in situ and in vitro, using the human term placenta as a model of physiological angiogenesis. Angiogenin was immunodetected by light and transmission electron microscopy, and its cellular distribution was established by double immunolabelling with cell markers including von Willebrand factor, platelet/endothelial cell adhesion molecule-1 (PECAM-1), CD34, Tie-2, vascular endothelial cadherin (VE-cadherin), vascular endothelial growth factor receptor-2 (VEGF-R2), erythropoeitin receptor (Epo-R), alpha-smooth muscle actin, CD45, cytokeratin 7, and Ki-67. Angiogenin immunoreactivity was detected in villous and extravillous trophoblasts, the trophoblast basement membrane, the endothelial basal lamina, foetal blood vessels, foetal and maternal red blood cells, and amnionic cells. Its expression was confirmed by in situ hybridisation with a digoxygenin-labelled cDNA probe and reverse transcriptase-polymerase chain reaction amplification. Villous cytotrophoblasts, isolated and differentiated in vitro into a functional syncytiotrophoblast, expressed and secreted angiogenin. Given its known biological activities in vitro and its observed pattern of expression, these data suggest that, in human placenta, angiogenin has a role not only in angiogenesis but also in vascular and tissue homeostasis, maternal immune tolerance of the foetus, and host defences.
Published: 2004

45. Role of nitric oxide synthases in the infarct size-reducing effect conferred by heat stress in isolated rat hearts

Author: Arnaud, Claire, Laubriet, Aline, Joyeux, Marie, Godin-Ribuot, Diane, Rochette, Luc, Demenge, Pierre, Ribuot, Christophe, Hypoxie : Physiopathologie Respiratoire et Cardiovasculaire (HP2), Université Joseph Fourier - Grenoble 1 (UJF)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Physiopathologie et de Pharmacologie Cardiovasculaire Expérimentale (LPPCE), Université de Bourgogne (UB), Hypoxie : Physiopathologie Respiratoire et Cardiovasculaire ( HP2 ), Université Joseph Fourier - Grenoble 1 ( UJF ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Laboratoire de Physiopathologie et de Pharmacologie Cardiovasculaire Expérimentale ( LPPCE ), Université de Bourgogne ( UB ), and Arnaud, Claire
Subjects: Male, MESH : Immunohistochemistry, Myocardial Infarction, Nitric Oxide Synthase Type II, HSP72 Heat-Shock Proteins, MESH: Heat-Shock Proteins, Infarct size, Heat stress, MESH : Heat Stress Disorders, MESH : Myocardium, MESH: Animals, Enzyme Inhibitors, MESH: Heat Stress Disorders, Heat-Shock Proteins, MESH : Isoenzymes, MESH : Hemodynamics, MESH : Rats, [ SDV.MHEP.CSC ] Life Sciences [q-bio]/Human health and pathology/Cardiology and cardiovascular system, Immunohistochemistry, MESH: Myocardial Reperfusion Injury, [SDV.MHEP.CSC] Life Sciences [q-bio]/Human health and pathology/Cardiology and cardiovascular system, Isoenzymes, MESH: Myocardial Infarction, MESH: Enzyme Inhibitors, MESH : Lysine, Papers, Heat stress protein, MESH: Isoenzymes, MESH : HSP72 Heat-Shock Proteins, MESH: Nitric Oxide Synthase, MESH: Nitric Oxide Synthase Type II, MESH: Hemodynamics, MESH: Myocardium, MESH: Rats, MESH : Male, MESH : Myocardial Reperfusion Injury, Myocardial Reperfusion Injury, In Vitro Techniques, Heat Stress Disorders, MESH : Rats, Wistar, MESH: HSP72 Heat-Shock Proteins, [SDV.MHEP.CSC]Life Sciences [q-bio]/Human health and pathology/Cardiology and cardiovascular system, Animals, MESH: Lysine, Rats, Wistar, MESH : Enzyme Inhibitors, Lysine, Myocardium, Nitric oxide synthases, Hemodynamics, MESH : Nitric Oxide Synthase, MESH: Immunohistochemistry, MESH: Rats, Wistar, MESH : Nitric Oxide Synthase Type II, MESH : Heat-Shock Proteins, MESH: Male, Rats, MESH : Animals, Nitric Oxide Synthase, MESH : Myocardial Infarction
Abstract: International audience; Nitric oxide (NO) donors are known to induce both delayed cardioprotection and myocardial heat stress protein (HSP) expression. Moreover, heat stress (HS), which also protects myocardium against ischaemic damages, is associated with a NO release. Therefore, we have investigated the implication of NO in HS-induced resistance to myocardial infarction, in the isolated rat heart model. Rats were divided in six groups (n=10 in each group), subjected or not to heat stress (42 degrees C internal temperature, 15 min) and treated or not with nitro-L-arginine-methylester (L-NAME) a non-selective inhibitor of NO synthase isoforms, or L-N(6)-(1-imino-ethyl)lysine (L-NIL), a selective inhibitor of the inducible NO synthase. Twenty-four hours after heat stress, their hearts were isolated, retrogradely perfused, and subjected to a 30-min occlusion of the left coronary artery followed by 120 min of reperfusion. Infarct-to-risk ratio was significantly reduced in HS (18.7+/-1.6%) compared to Sham (33.0+/-1.7%) hearts. This effect was abolished in L-NAME-treated (41.7+/-3.1% in HS+L-NAME vs 35.2+/-3.0% in Sham+L-NAME ) and L-NIL-treated (36.1+/-3.4% in HS+L-NIL vs 42.1+/-4.6% in Sham+L-NIL) groups. Immunohistochemical analysis of myocardial HSP 27 and 72 showed an HS-induced increase of these proteins, which was not modified by L-NAME pretreatment. We conclude that NO synthases, and in particular the inducible isoform, appear to play a role in the heat stress-induced cardioprotection, independently of HSP 27 and 72 levels. Further investigations are required to elucidate the precise role of HSPs in this adaptive response.
Published: 2001

46. Missense Mutations in SLC26A8, Encoding a Sperm-Specific Activator of CFTR, Are Associated with Human Asthenozoospermia

Author: Thierry Bienvenu, Pierre F. Ray, Nathalie Da Silva, Mathilde Jollivet, Aminata Touré, Pierre Tufféry, Baptiste Rode, Denise Escalier, Gérard Gacon, Emmanuel Dulioust, Frédéric Becq, Jean-Philippe Wolf, Caroline Norez, Thassadite Dirami, Natacha Gaitch, Institut Cochin (IC UM3 (UMR 8104 / U1016)), 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 physiologie et biologie cellulaires (IPBC), Université de Poitiers-Centre National de la Recherche Scientifique (CNRS), Physiopathologie des maladies génétiques d'expression pédiatrique, Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Biochimie et Génétique Moléculaire, Université Paris Descartes - Paris 5 (UPD5), Molécules Thérapeutiques in silico (MTI), Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM), Service d'Histologie-Embryologie, Biologie de la Reproduction (CECOS Paris Cochin), Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), AGeing and IMagery (AGIM), Centre National de la Recherche Scientifique (CNRS)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Joseph Fourier - Grenoble 1 (UJF)-Université Pierre Mendès France - Grenoble 2 (UPMF), Université Joseph Fourier - Grenoble 1 (UJF), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Université Pierre Mendès France - Grenoble 2 (UPMF)-Université Joseph Fourier - Grenoble 1 (UJF)-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), Morillon, Christelle, Université Pierre Mendès France - Grenoble 2 (UPMF)-Université Joseph Fourier - Grenoble 1 (UJF)-École Pratique des Hautes Études (EPHE), Institut Cochin ( UM3 (UMR 8104 / U1016) ), 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 physiologie et biologie cellulaires ( IPBC ), Université de Poitiers-Centre National de la Recherche Scientifique ( CNRS ), Institut National de la Santé et de la Recherche Médicale ( INSERM ) -Université Pierre et Marie Curie - Paris 6 ( UPMC ), Université Paris Descartes - Paris 5 ( UPD5 ), Molécules Thérapeutique in silico, Recherche de molécules à visée thérapeutique par approches In Silico ( MTI ), Université Paris Diderot - Paris 7 ( UPD7 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Service d'Histologie-Embryologie, Biologie de la Reproduction ( CECOS ), Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Cochin [AP-HP], AGeing and IMagery ( AGIM ), Université Pierre Mendès France - Grenoble 2 ( UPMF ) -Université Joseph Fourier - Grenoble 1 ( UJF ) -École pratique des hautes études ( EPHE ) -Centre National de la Recherche Scientifique ( CNRS ), and Université Joseph Fourier - Grenoble 1 ( UJF )
Subjects: Male, MESH : Immunohistochemistry, [SDV]Life Sciences [q-bio], MESH: Cricetinae, Cystic Fibrosis Transmembrane Conductance Regulator, Antiporters, MESH : Antiporters, 0302 clinical medicine, MESH: Cricetulus, Cricetinae, MESH : CHO Cells, Missense mutation, Genetics(clinical), MESH: Animals, MESH : Immunoblotting, Genetics (clinical), Sperm motility, Genetics, MESH: Cystic Fibrosis Transmembrane Conductance Regulator, 0303 health sciences, 030219 obstetrics & reproductive medicine, MESH: Immunoblotting, Chinese hamster ovary cell, MESH: Spermatozoa, MESH : Cricetinae, MESH: Genetic Predisposition to Disease, MESH: Anion Transport Proteins, MESH: Sperm Motility, Immunohistochemistry, Spermatozoa, Cystic fibrosis transmembrane conductance regulator, Transport protein, [SDV] Life Sciences [q-bio], Sulfate Transporters, Asthenozoospermia, MESH: Sperm Capacitation, Sperm Motility, MESH : Asthenozoospermia, MESH : Sperm Capacitation, MESH : Male, Urology, MESH : Cricetulus, Immunoblotting, Anion Transport Proteins, Mutation, Missense, CHO Cells, Biology, 03 medical and health sciences, Cricetulus, MESH: CHO Cells, Capacitation, Report, MESH : Spermatozoa, medicine, Animals, Humans, Genetic Predisposition to Disease, 030304 developmental biology, MESH: Mutation, Missense, MESH: Humans, [ SDV ] Life Sciences [q-bio], business.industry, Activator (genetics), MESH : Cystic Fibrosis Transmembrane Conductance Regulator, MESH : Humans, MESH: Immunohistochemistry, medicine.disease, Sperm, Molecular biology, MESH: Male, MESH: Asthenozoospermia, MESH : Sperm Motility, MESH : Anion Transport Proteins, biology.protein, MESH : Genetic Predisposition to Disease, MESH: Antiporters, MESH : Animals, business, Sperm Capacitation, MESH : Mutation, Missense
Abstract: International audience; The cystic fibrosis transmembrane conductance regulator (CFTR) is present in mature sperm and is required for sperm motility and capacitation. Both these processes are controlled by ions fluxes and are essential for fertilization. We have shown that SLC26A8, a sperm-specific member of the SLC26 family of anion exchangers, associates with the CFTR channel and strongly stimulates its activity. This suggests that the two proteins cooperate to regulate the anion fluxes required for correct sperm motility and capacitation. Here, we report on three heterozygous SLC26A8 missense mutations identified in a cohort of 146 men presenting with asthenozoospermia: c.260G>A (p.Arg87Gln), c.2434G>A (p.Glu812Lys), and c.2860C>T (p.Arg954Cys). These mutations were not present in 121 controls matched for ethnicity, and statistical analysis on a control population of 8,600 individuals (from dbSNP and 1000 Genomes) showed them to be associated with asthenozoospermia with a power > 95%. By cotransfecting Chinese hamster ovary (CHO)-K1 cells with SLC26A8 variants and CFTR, we showed that the physical interaction between the two proteins was partly conserved but that the capacity to activate CFTR-dependent anion transport was completely abolished for all mutants. Biochemical studies revealed the presence of much smaller amounts of protein for all variants, but these amounts were restored to wild-type levels upon treatment with the proteasome inhibitor MG132. Immunocytochemistry also showed the amounts of SLC26A8 in sperm to be abnormally small in individuals carrying the mutations. These mutations might therefore impair formation of the SLC26A8-CFTR complex, principally by affecting SLC26A8 stability, consistent with an impairment of CFTR-dependent sperm-activation events in affected individuals.
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