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7 results on '"MESH: Fixatives"'

1. Detection of the TMPRSS2-ETS fusion gene in prostate carcinomas: retrospective analysis of 55 formalin-fixed and paraffin-embedded samples with clinical data

Author

Cécile Rouzier, Jean Amiel, Jean-François Michiels, Laure Valério, Juliette Haudebourg, Florence Pedeutour, Xavier Carpentier, Laboratory of Solid Tumors Genetics, Nice University Hospital, 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), Laboratoire d'Anatomo-Pathologie, Hôpital Pasteur [Nice] (CHU), Laboratoire d'Urologie, Department of Public Health, and Hôpital l'Archet

Subjects
Male, Cancer Research, Oncogene Proteins, Fusion, Oncogene Proteins, MESH: Paraffin Embedding, Chromosomal translocation, MESH: Formaldehyde, law.invention, Fusion gene, MESH: Aged, 80 and over, 0302 clinical medicine, law, Prostate, MESH: Reverse Transcriptase Polymerase Chain Reaction, MESH: Fixatives, MESH: Serine Endopeptidases, Polymerase chain reaction, MESH: Aged, Aged, 80 and over, 0303 health sciences, MESH: Middle Aged, Paraffin Embedding, Reverse Transcriptase Polymerase Chain Reaction, Serine Endopeptidases, MESH: Transcription Factors, Middle Aged, 3. Good health, DNA-Binding Proteins, medicine.anatomical_structure, 030220 oncology & carcinogenesis, Adenocarcinoma, Adenovirus E1A Proteins, PCA3, MESH: Proto-Oncogene Proteins c-ets, [SDV.CAN]Life Sciences [q-bio]/Cancer, Biology, Fixatives, 03 medical and health sciences, Antigen, Formaldehyde, Proto-Oncogene Proteins, Genetics, medicine, Humans, Molecular Biology, Aged, Retrospective Studies, 030304 developmental biology, MESH: Humans, Proto-Oncogene Proteins c-ets, MESH: Adenocarcinoma, Prostatic Neoplasms, MESH: Retrospective Studies, medicine.disease, MESH: Adenovirus E1A Proteins, MESH: Male, MESH: Proto-Oncogene Proteins, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, MESH: Prostatic Neoplasms, Immunology, Cancer research, Feasibility Studies, MESH: Feasibility Studies, MESH: DNA-Binding Proteins, MESH: Oncogene Proteins, Fusion, Transcription Factors
Abstract

The recent identification of fusion genes involving ETS family members in human prostate adenocarcinoma has confirmed the hypothesis that recurrent specific aberrations such as fusion genes may be as frequent in epithelial tumors as they are in leukemias and sarcomas. However, reciprocal translocations with fusion genes are often not detectable in carcinomas by conventional karyotyping because of additional complex chromosomal abnormalities. We retrospectively analyzed a large series of formalin-fixed, paraffin-embedded samples including 55 prostate carcinomas and 11 benign prostate tumors. We identified the fusion gene TMPRSS2-ERG by reverse-transcriptase polymerase chain reaction (RT-PCR) in 40/55 carcinomas (72%). Our study demonstrates that the detection of ETS fusion gene by RT-PCR is feasible on formalin-fixed and paraffin-embedded samples. No significant association between the presence of the fusion gene and any clinical feature, such as preoperative serum prostate-specific antigen (PSA) level (PSA>20 or PSA< or =20), pTNM stage including capsule invasion, seminal vesicle invasion, and lymph nodes metastases, or recurrence was observed in our series.

Published
2008
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2. Correlative fluorescence and electron microscopy of biocytin-filled neurons with a preservation of the postsynaptic ultrastructure

Author

Yehezkel Ben-Ari, Youri Morozov, Ilgam Khalilov, Alfonso Represa, Epilepsie et ischémie cérébrale, Université de la Méditerranée - Aix-Marseille 2-Institut National de la Santé et de la Recherche Médicale (INSERM), and Tyzio, Roman

Subjects
MESH: Hippocampus, Tissue Fixation, MESH: Neurons, MESH: Microscopy, Fluorescence, MESH: Streptavidin, Hippocampus, MESH: Animals, Newborn, law.invention, chemistry.chemical_compound, 0302 clinical medicine, law, Postsynaptic potential, Fluorescence microscope, MESH: Animals, MESH: Fixatives, MESH: Tissue Embedding, Neurons, 0303 health sciences, Pyramidal Cells, General Neuroscience, MESH: Fluorescent Dyes, MESH: Interneurons, Immunohistochemistry, MESH: Neuroanatomy, medicine.anatomical_structure, MESH: Axons, MESH: Rats, Synaptic Membranes, MESH: Microscopy, Electron, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, Biology, MESH: Dendrites, Fixatives, 03 medical and health sciences, Organ Culture Techniques, Interneurons, Biocytin, medicine, Animals, MESH: Lysine, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, Fluorescent Dyes, 030304 developmental biology, Organelles, Tissue Embedding, Lysine, MESH: Immunohistochemistry, MESH: Pyramidal Cells, Dendrites, MESH: Synaptic Membranes, MESH: Organ Culture Techniques, Axons, Rats, Microscopy, Electron, Neuroanatomy, Animals, Newborn, Microscopy, Fluorescence, nervous system, chemistry, Osmium tetroxide, Ultrastructure, Biophysics, Streptavidin, Neuron, MESH: Tissue Fixation, Electron microscope, Neuroscience, Postsynaptic density, 030217 neurology & neurosurgery, MESH: Organelles
Abstract

International audience; Several techniques enable to inject intracellularly neurons with dyes and to use light and electron microscopy to correlate the physiological data with the morphological properties of the neuron. However, the ultrastructure of the neuron is usually obscured by the injected dye thus notably precluding the analysis of the postsynaptic specialisation and that of the other organelles. To overcome this problem, we have developed a technique based on fluorophore- and ultra small gold-conjugated streptavidins. We report, that this method facilitates the identification of intracellular organelles of the biocytin-filled neuron and of postsynaptic densities. This method is valid for the study of early postnatal neurons that are particularly refractory to this type of analysis. The procedure introduced here consists of the following steps: (1) injection of biocytin into the neuron by a patch-clamp pipette, (2) aldehyde fixation, (3) reaction with a fluorophore-conjugated streptavidin, (4) analysis with a fluorescence microscope, (5) formation of avidin-biotin complexes (ABC), (6) reaction with an ultra small gold-conjugated streptavidin, (7) silver enhancement of gold, (8) postfixation with osmium tetroxide and embedding in resin, (9) ultrathin sectioning and analysis with an electron microscope. Using this method, we show that in early postnatal hippocampal neurons, that have been injected with biocytine, it is possible to determine the morphology of the dendritic and axonal trees (including very thin details such as spines and filopodia) and to identify the localisation of the symmetric and asymmetric synapses on dendrites of the injected neuron.

Published
2002
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3. Performance and cost efficiency of KRAS mutation testing for metastatic colorectal cancer in routine diagnosis: the MOKAECM study, a nationwide experience

Author

Pierre Laurent-Puig, Jean-François Emile, Marie-Claude Gorisse, Pascaline Aucouturier, Jean-Philippe Merlio, Florence de Fraipont, Jean-Louis Merlin, Isabelle Durand-Zaleski, Frédérique Nowak, Karen Leroy, Thierry Frebourg, Alain Morel, Hélène Blons, Delphine Le Corre, Gilles Chatellier, Jean-François Côté, Jean-Christophe Sabourin, L'Houcine Ouafik, Etienne Rouleau, Jean-Christophe Boyer, Nathanaël Charrier, Jean-Christophe Pages, Gérard Milano, Patricia de Cremoux, Bases moléculaires de la réponse aux xénobiotiques (U775 (IFR95)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Oncogénétique, CRLCC René Huguenin, Service de Santé Publique, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Hôpital Albert Chenevier, Centre d'Investigation Clinique - Epidemiologie Clinique/essais Cliniques [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)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Recherches Cliniques de Montréal (IRCM), Université de Montréal (UdeM), Virus, pseudo-virus: Morphogénèse et Antigénicité, Université de Tours (UT)-EA3856, Hôpital Bretonneau, Centre Hospitalier Régional Universitaire de Tours (CHRU Tours), Institut d'oncologie/développement Albert Bonniot de Grenoble (INSERM U823), Université Joseph Fourier - Grenoble 1 (UJF)-CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Biochimie [CHRU Nîmes], Centre Hospitalier Universitaire de Nîmes (CHU Nîmes), CHU Bordeaux [Bordeaux], Oppelia, Institut Jean Godinot [Reims], UNICANCER, Hôpital Saint Louis, Hopital Saint-Louis [AP-HP] (AP-HP), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Département de pathologie [Mondor], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Département de pharmacogénomique, Centre de Lutte contre le Cancer Antoine Lacassagne [Nice] (UNICANCER/CAL), UNICANCER-Université Côte d'Azur (UCA)-UNICANCER-Université Côte d'Azur (UCA), 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), Centre de Recherche en Automatique de Nancy (CRAN), Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), Centre Alexis Vautrin (CAV), CHU Rouen, Normandie Université (NU), Institut national du cancer [Boulogne] (INCA), Génétique du cancer et des maladies neuropsychiatriques (GMFC), Université de Rouen Normandie (UNIROUEN), Normandie Université (NU)-Normandie Université (NU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Biomarqueurs et essais cliniques en Cancérologie et Onco-Hématologie (BECCOH), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Université Paris-Saclay, Service d'anatomie pathologique [CHU Ambroise-Paré], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Ambroise Paré [AP-HP], Département de la recherche clinique et du développement, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôtel-Dieu-Unité de Recherche Clinique en Economie de la santé d'Ile de France (URC Eco), Médecine Personnalisée, Pharmacogénomique, Optimisation Thérapeutique (MEPPOT - U1147), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Tours-EA3856, Institut National de la Santé et de la Recherche Médicale (INSERM)-EFS-CHU Grenoble-Université Joseph Fourier - Grenoble 1 (UJF), Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Henri Mondor-Hôpital Albert Chenevier, 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)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Recherches Cliniques de Montréal, Université de Montréal, Université de Montréal [Montréal], Biochimie et Physiologie Moléculaire des Plantes (BPMP), Université de Montpellier (UM)-Centre international d'études supérieures en sciences agronomiques (Montpellier SupAgro)-Institut national d’études supérieures agronomiques de Montpellier (Montpellier SupAgro)-Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Institut Jean Godinot, CRLCC Jean Godinot, CHU Saint Louis [APHP], Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), CRLCC Antoine Lacassagne, Laboratoire épidémiologie et oncogénèse des tumeurs digestives, Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Service d'anatomie pathologique, Université de Versailles Saint-Quentin-en-Yvelines (UVSQ)-Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-Hôpital Ambroise Paré, Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Cochin [AP-HP]-Hôtel-Dieu-Unité de Recherche Clinique en Economie de la santé d'Ile de France (URC Eco), Centre National de la Recherche Scientifique (CNRS)-Université de Lorraine (UL), Bases moléculaires de la réponse aux xénobiotiques ( U775 (IFR95) ), 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)-Hôpital Henri Mondor-Hôpital Albert Chenevier, Centre d'Investigation Clinique - Epidemiologie Clinique/essais Cliniques de l'Hopital Europeen Georges Pompidou, Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Européen Georges Pompidou [APHP] ( HEGP ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université de Montréal, CHRU Tours, Institut d'oncologie/développement Albert Bonniot de Grenoble ( INSERM U823 ), Université Joseph Fourier - Grenoble 1 ( UJF ) -CHU Grenoble-EFS-Institut National de la Santé et de la Recherche Médicale ( INSERM ), Biochimie et Physiologie Moléculaire des Plantes ( BPMP ), Centre National de la Recherche Scientifique ( CNRS ) -Université de Montpellier ( UM ) -Centre international d'études supérieures en sciences agronomiques ( Montpellier SupAgro ) -Institut national d’études supérieures agronomiques de Montpellier ( Montpellier SupAgro ) -Institut national de la recherche agronomique [Montpellier] ( INRA Montpellier ), Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 ( UPEC UP12 ), 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 ), Centre de Recherche en Automatique de Nancy ( CRAN ), Université de Lorraine ( UL ) -Centre National de la Recherche Scientifique ( CNRS ), Centre Alexis Vautrin ( CAV ), Institut national du cancer [Boulogne] ( INCA ), Génétique du cancer et des maladies neuropsychiatriques ( GMFC ), Université de Rouen Normandie ( UNIROUEN ), Normandie Université ( NU ) -Normandie Université ( NU ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), Université de Versailles Saint-Quentin-en-Yvelines ( UVSQ ), Université de Versailles Saint-Quentin-en-Yvelines ( UVSQ ) -Assistance publique - Hôpitaux de Paris (AP-HP)-Hôpital Ambroise Paré, Assistance publique - Hôpitaux de Paris (AP-HP)-CHU Cochin [AP-HP]-Hôtel-Dieu-Unité de Recherche Clinique en Economie de la santé d'Ile de France (URC Eco), Médecine Personnalisée, Pharmacogénomique, Optimisation Thérapeutique ( MEPPOT - U1147 ), Université Paris Descartes - Paris 5 ( UPD5 ) -Institut National de la Santé et de la Recherche Médicale ( INSERM ), and Merlin, Jean-Louis

Subjects
Oncology, Colorectal cancer, Epidemiology, Molecular oncology, MESH: Antibodies, Monoclonal, 0302 clinical medicine, MESH : Fixatives, MESH: Fixatives, Neoplasm Metastasis, lcsh:Science, MESH: Tissue Embedding, 0303 health sciences, MESH: Genetic Testing, Antibodies, Monoclonal, 3. Good health, MESH: Reproducibility of Results, ErbB Receptors, [ SDV.BBM.GTP ] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], 030220 oncology & carcinogenesis, MESH : Neoplasm Metastasis, MESH : Proto-Oncogene Proteins, [SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], Medicine, MESH : Colorectal Neoplasms, MESH : Sensitivity and Specificity, MESH: ras Proteins, medicine.medical_specialty, MESH: Receptor, Epidermal Growth Factor, Gastroenterology and Hepatology, Sensitivity and Specificity, Proto-Oncogene Proteins p21(ras), 03 medical and health sciences, Fixatives, MESH : Genetic Testing, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], External quality assessment, Gastrointestinal Tumors, Cancer Detection and Diagnosis, Humans, Genetic Testing, Genotyping, Biology, MESH: Humans, Tissue Embedding, MESH : Reproducibility of Results, MESH : Humans, lcsh:R, medicine.disease, MESH: Neoplasm Metastasis, MESH: Sensitivity and Specificity, MESH : Biopsy, ras Proteins, lcsh:Q, MESH : Sequence Analysis, DNA, MESH: Sequence Analysis, DNA, Biopsy, Cancer Treatment, lcsh:Medicine, Bioinformatics, medicine.disease_cause, MESH : Tissue Embedding, MESH: Biopsy, MESH : Receptor, Epidermal Growth Factor, Gastrointestinal Cancers, Sanger sequencing, Multidisciplinary, Drug development, MESH : Antibodies, Monoclonal, symbols, KRAS, France, Colorectal Neoplasms, Research Article, MESH: Cell Line, Tumor, Clinical Research Design, Rectal Cancer, symbols.namesake, Antibody Therapy, Diagnostic Medicine, Molecular genetics, Internal medicine, Cell Line, Tumor, Proto-Oncogene Proteins, medicine, MESH : France, 030304 developmental biology, Population Biology, MESH : Cell Line, Tumor, business.industry, Reproducibility of Results, Cancers and Neoplasms, Sequence Analysis, DNA, MESH: France, MESH: Proto-Oncogene Proteins, Gastric Cancer, MESH : ras Proteins, business, MESH: Colorectal Neoplasms
Abstract

International audience; PURPOSE: Rapid advances in the understanding of cancer biology have transformed drug development thus leading to the approval of targeted therapies and to the development of molecular tests to select patients that will respond to treatments. KRAS status has emerged as a negative predictor of clinical benefit from anti-EGFR antibodies in colorectal cancer, and anti-EGFR antibodies use was limited to KRAS wild type tumors. In order to ensure wide access to tumor molecular profiling, the French National Cancer Institute (INCa) has set up a national network of 28 regional molecular genetics centers. Concurrently, a nationwide external quality assessment for KRAS testing (MOKAECM) was granted to analyze reproducibility and costs. METHODS: 96 cell-line DNAs and 24 DNA samples from paraffin embedded tumor tissues were sent to 40 French laboratories. A total of 5448 KRAS results were collected and analyzed and a micro-costing study was performed on sites for 5 common methods by an independent team of health economists. RESULTS: This work provided a baseline picture of the accuracy and reliability of KRAS analysis in routine testing conditions at a nationwide level. Inter-laboratory Kappa values were >0.8 for KRAS results despite differences detection methods and the use of in-house technologies. Specificity was excellent with only one false positive in 1128 FFPE data, and sensitivity was higher for targeted techniques as compared to Sanger sequencing based methods that were dependent upon local expertise. Estimated reagent costs per patient ranged from €5.5 to €19.0. CONCLUSION: The INCa has set-up a network of public laboratories dedicated to molecular oncology tests. Our results showed almost perfect agreements in KRAS testing at a nationwide level despite different testing methods ensuring a cost-effective equal access to personalized colorectal cancer treatment.

Published
2013
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4. 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
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5. CEMOVIS on a pathogen: analysis of Bacillus anthracis spores

Author

Evelyne Couture-Tosi, Georges Michel Haustant, Martin Sachse, Gérard Pehau-Arnaudet, Jean-Luc Ranck, Toxines et Pathogénie Bactérienne, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Biologie structurale et agents infectieux (BSAI), Institut Pasteur [Paris] (IP), Centre National de la Recherche Scientifique (CNRS), Imagopole (CITECH), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris], and CNRS

Subjects
Bacillus cereus, Bacillus, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, spore, Microbiology, Anthrax, Fixatives, 03 medical and health sciences, MESH: Spores, Bacterial, MESH: Anthrax, MESH: Fixatives, MESH: Glutaral, 030304 developmental biology, Spores, Bacterial, 0303 health sciences, biology, 030306 microbiology, Cryoelectron Microscopy, fungi, Exosporium, Cell Biology, General Medicine, Cryo-Electron Microscopy of Vitreous Sections (CEMOVIS), biology.organism_classification, Vitrification, Spore, Bacillus anthracis, MESH: Bacillus anthracis, germination, MESH: Bacillus cereus, Glutaral, Germination, MESH: Vitrification, Ultrastructure, Bacterial spore, MESH: Cryoelectron Microscopy
Abstract

Background information. Under conditions of starvation, bacteria of Bacillus ssp. are able to form a highly structured cell type, the dormant spore. When the environment presents more favourable conditions, the spore starts to germinate, which will lead to the release of the vegetative form in the life cycle, the bacillus. For Bacillus anthracis, the aetiological agent of anthrax, germination is normally linked to host uptake and represents an important step in the onset of anthrax disease. Morphological studies analysing the organization of the spore and the changes during germination at the electron microscopy level were only previously performed with techniques relying on fixation with aldehydes and osmium, and subsequent dehydration, which can produce artefacts. Results and conclusions. In the present study, we describe the morphology of dormant spores using CEMOVIS (Cryo-Electron Microscopy of Vitreous Sections). Biosafety measures do not permit freezing of native spores of B. anthracis without chemical fixation. To study the influence of aldehyde fixation on the ultrastructure of the spore, we chose to analyse spores of the closely related non-pathogen Bacillus cereus T. For none of the investigated structures could we find a difference in morphology induced by aldehyde fixation compared with the native preparations for CEMOVIS. This result legitimizes work with aldehyde-fixed spores from B. anthracis. Using CEMOVIS, we describe two new structures present in the spore: a rectangular structure, which connects the BclA filaments with the basal layer of the exosporium, and a repetitive structure, which can be found in the terminal layer of the coat. We studied the morphological changes of the spore during germination. After outgrowth of the bacillus, coat and exosporium stay associated, and the layered organization of the coat, as well as the repetitive structure within it, remain unchanged.

Published
2010
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6. [Management of muscle and nerve biopsies: expert guidelines from two French professional societies, Société française de myologie et de l'Association française contre les myopathies] : Prise en charge des biopsies musculaires et nerveuses. Recommandations formalisées d'experts sous l'égide de la Société française de neuropathologie, de la Société française de myologie et de l'Association française contre les myopathies

Author

Uro-Soste, E., Fernandez, C., Authier, François-Jérôme, Bassez, G., Butori, C., Chapon, F., Delisle, M.-B., Dubourg, O., Feasson, L., Gherardi, R., Lacroix, C., Laquerriere, A., Letournel, F., Magy, Laurent, Maisonobe, T., Marcorelles, Pascale, Maurage, C.-A., Mezin, P., Mussini, J.-M., Penisson-Besnier, I., Romero, N.-B., Streichenberger, N., Vallat, Jean-Michel, Viennet, G., Vital, A., Voit, T., Boucharef, W., Figarella-Branger, D., Renseigné, Non, Service d'anatomie pathologique et histologie-cytologie [Rangueil], Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-CHU Toulouse [Toulouse]-Hôpital de Rangueil, CHU Toulouse [Toulouse], Centre de référence des maladies rares neuromusculaires Aquitaine-Grand Sud Ouest, Service d'anatomie pathologique et de neurophatologie, Assistance Publique - Hôpitaux de Marseille (APHM)-CHU Marseille, Centre de référence des maladies rares neuromusculaires, INSERM U955, équipe 10, Département de pathologie [Mondor], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Centre de référence des maladies rares neuromusculaires, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Institut Mondor de Recherche Biomédicale (IMRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12)-Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Henri Mondor, Institut Mondor de Recherche Biomédicale (IMRB), Institut National de la Santé et de la Recherche Médicale (INSERM)-IFR10-Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Laboratoire de pathologie clinique et expérimentale, Centre Hospitalier Universitaire de Nice (CHU Nice), Service de Neurologie [CHU Caen], Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), Centre de compétences pathologies neuromusculaires [CHU Caen], Thérapie des maladies du muscle strié, 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), 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), Service de physiologie clinique, unité de myologie, CHU Saint-Etienne, Centre de référence des neuropathies amyloïdes et autres neuropathies rares, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Bicêtre, Laboratoire de neuropathologie, Service d'Anatomie et Cytologie Pathologique [CHU Rouen], CHU Rouen, Normandie Université (NU)-Normandie Université (NU)-Université de Rouen Normandie (UNIROUEN), Normandie Université (NU), Département de Pathologie Cellulaire et Tissulaire, Centre Hospitalier Universitaire d'Angers (CHU Angers), PRES Université Nantes Angers Le Mans (UNAM)-PRES Université Nantes Angers Le Mans (UNAM), Laboratoire de Neurobiologie et Neuropathologie, Service de Neurologie [CHU Limoges], CHU Limoges, Biomolécules Thérapies anti-tumorales (EA4021), Université de Limoges (UNILIM)-Génomique, Environnement, Immunité, Santé, Thérapeutique (GEIST FR CNRS 3503), Centre de référence national neuropathies périphériques rares [CHU Limoges], Service d'Anatomomie pathologique, Centre Hospitalier Régional Universitaire de Brest (CHRU Brest), Centre de compétence des maladies rares neuromusculaires, Service d'anatomie pathologique, Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Département d'anatomie et cythologie pathologique, CHU Grenoble-Hôpital Michallon, CHU Grenoble, Service de médecine interne [Nantes], Université de Nantes (UN)-Hôtel-Dieu-Centre hospitalier universitaire de Nantes (CHU Nantes), Service de neurologie [Angers], Université d'Angers (UA)-Centre Hospitalier Universitaire d'Angers (CHU Angers), Service de pathologie et neuropatologie, Hospices Civils de Lyon (HCL), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon, Estrogènes, Expression génique et pathologies du Système Nerveux Central - UFC (E2SNC / ESTROGENES), 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), Service de pathologie [Bordeaux], Université Bordeaux Segalen - Bordeaux 2-CHU Bordeaux [Bordeaux]-Groupe hospitalier Pellegrin, CHU Bordeaux [Bordeaux], Service des actions médicales, paramédicales et psychologiques, Association Française contre les Myopathies, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-CHU Toulouse [Toulouse], IMRB - 'Biologie du système neuromusculaire' [Créteil] (U955 Inserm - UPEC), École nationale vétérinaire - Alfort (ENVA)-Institut Mondor de Recherche Biomédicale (IMRB), Centre de référence des maladies rares neuromusculaires [CHU Pitié-Salpétriêre], 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), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Centre de Reference des Maladies Neuromusculaires - Atlantique Occitanie Caraïbe (CRMN - AOC), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), and Centre Hospitalier Universitaire de Saint-Etienne [CHU Saint-Etienne] (CHU ST-E)

Subjects
MESH: Muscle, Skeletal, [SDV.GEN]Life Sciences [q-bio]/Genetics, MESH: Humans, [SDV.NEU.NB]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC]/Neurobiology, MESH: Paraffin Embedding, MESH: Immunohistochemistry, [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: Peripheral Nerves, MESH: Biopsy, [SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics, [SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN], MESH: Fixatives, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], [SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology, MESH: Glutaral, ComputingMilieux_MISCELLANEOUS
Abstract

International audience

Published
2010
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7. Usefulness of molecular biology performed with formaldehyde-fixed paraffin embedded tissue for the diagnosis of combined pulmonary invasive mucormycosis and aspergillosis in an immunocompromised patient

Author

Véronique Hofman, Michele Baumann, Nicolas Venissac, Dea Garcia-Hermoso, Paul Hofman, Martine Gari-Toussaint, Bernard Padovani, Abdelmajid Dhouibi, Catherine Butori, Gieri Cathomas, Laboratory of Clinical and Experimental Pathology, Louis Pasteur Hospital, Human Biobank, Department of Radiology, Laboratory of Mycology, Archet II Hospital, Centre National de Référence Mycologie et Antifongiques-Mycologie Moléculaire (CNRMA), Institut Pasteur [Paris], Laboratory for Molecular and Infectious Disease Pathology, Cantonal Institute for Pathology, Department of Thoracic Surgery, and Institut Pasteur [Paris] (IP)

Subjects
Male, MESH: Fatal Outcome, Pathology, Tissue Fixation, MESH: Paraffin Embedding, Case Report, MESH: Formaldehyde, Aspergillosis, Polymerase Chain Reaction, MESH: Kidney Transplantation, Aspergillus fumigatus, Fatal Outcome, 0302 clinical medicine, MESH: Rhizopus, MESH: Immunocompromised Host, MESH: Fixatives, Lung, MESH: Aged, Invasive Pulmonary Aspergillosis, 0303 health sciences, Paraffin Embedding, biology, General Medicine, MESH: Predictive Value of Tests, 3. Good health, MESH: Staining and Labeling, medicine.anatomical_structure, 030220 oncology & carcinogenesis, MESH: Immunosuppressive Agents, MESH: Aspergillus fumigatus, MESH: Tomography, X-Ray Computed, Bronchoalveolar Lavage Fluid, Immunosuppressive Agents, Rhizopus, medicine.drug, lcsh:RB1-214, Mucorales, MESH: Pneumonia, MESH: Invasive Pulmonary Aspergillosis, Rhizopus microsporus, medicine.medical_specialty, Histology, Pathology and Forensic Medicine, Fixatives, Immunocompromised Host, Necrosis, 03 medical and health sciences, Predictive Value of Tests, Formaldehyde, medicine, lcsh:Pathology, Humans, Mucormycosis, MESH: Lung, MESH: Mucormycosis, Aged, MESH: Necrosis, Voriconazole, MESH: Humans, Staining and Labeling, MESH: Bronchoalveolar Lavage Fluid, 030306 microbiology, MESH: Polymerase Chain Reaction, Pneumonia, biology.organism_classification, medicine.disease, Kidney Transplantation, MESH: Male, MESH: Tissue Fixation, Tomography, X-Ray Computed, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract

Immunocompromised patients who develop invasive filamentous mycotic infections can be efficiently treated if rapid identification of the causative fungus is obtained. We report a case of fatal necrotic pneumonia caused by combined pulmonary invasive mucormycosis and aspergillosis in a 66 year-old renal transplant recipient. Aspergillus was first identified during the course of the disease by cytological examination and culture (A. fumigatus) of bronchoalveolar fluid. Hyphae of Mucorales (Rhizopus microsporus) were subsequently identified by culture of a tissue specimen taken from the left inferior pulmonary lobe, which was surgically resected two days before the patient died. Histological analysis of the lung parenchyma showed the association of two different filamentous mycoses for which the morphological features were evocative of aspergillosis and mucormycosis. However, the definitive identification of the associative infection was made by polymerase chain reaction (PCR) performed on deparaffinized tissue sections using specific primers for aspergillosis and mucormycosis. This case demonstrates that discrepancies between histological, cytological and mycological analyses can occur in cases of combined mycotic infection. In this regard, it shows that PCR on selected paraffin blocks is a very powerful method for making or confirming the association of different filamentous mycoses and that this method should be made available to pathology laboratories.

Published
2010
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