Hospices Civils de Lyon 6 articles published in JoVE Biology Improved Enzyme Protection Assay to Study Staphylococcus aureus Internalization and Intracellular Efficacy of Antimicrobial Compounds Josselin Rigaill*1,2, Estelle Audoux*1, Killian Rodriguez1, Aurélien Peyron1, Philippe Berthelot1,3, Jérôme Josse4,5, Frédéric Laurent4,5, Robin Caire1, Paul O. Verhoeven1,2 1CIRI, Centre International de Recherche en Infectiologie, GIMAP team, University of Lyon, INSERM U1111, CNRS, UMR5308, ENS Lyon, UCBL1, University of St-Etienne, France, 2Department of Infectious Agents and Hygiene, University Hospital of St-Etienne, St-Etienne, France, 3Department of Infectious Diseases, University Hospital of St-Etienne, St-Etienne, France, 4CIRI, Centre International de Recherche en Infectiologie, Staphylococcal Pathogenesis team, University of Lyon, INSERM U1111, CNRS UMR5308, ENS Lyon, UCBL1, University of Lyon, Lyon, France, 5Department of Bacteriology, Institute for Infectious Agents, Hospices Civils de Lyon, Lyon, France This protocol aims to describe how to study the extent of Staphylococcus aureus internalization and its ability to survive inside the human host cell, as well as the intracellular efficacy of antimicrobial compounds. Cancer Research Semi-automatic PD-L1 Characterization and Enumeration of Circulating Tumor Cells from Non-small Cell Lung Cancer Patients by Immunofluorescence Jessica Garcia1,2,3, David Barthelemy1,2,3, Florence Geiguer1,2, Julie Ballandier1,2,3, Kathryn W. Li4, Jean-Phillippe Aurel4, Frédérique Le Breton5, Claire Rodriguez-Lafrasse1, Brigitte Manship3, Sébastien Couraud6,7, Léa Payen1,2,3 1Laboratoire de Biochimie et Biologie Moléculaire, Groupe Hospitalier Sud, Hospices Civils de Lyon, 2Circulating Cancer (CIRCAN) Program, Hospices Civils de Lyon Cancer Institute, 3University of Lyon, Claude Bernard University, Cancer Research Center of Lyon, 4Biolidics Limited, 5Institut de Pathologie Multisites des HCL-Site Sud, Hospices Civils de Lyon, 6Acute Respiratory Disease and Thoracic Oncology Department, Lyon Sud Hospital, Hospices Civils de Lyon Cancer Institute, 7EMR-3738 Therapeutic Targeting in Oncology, Lyon Sud Medical Faculty, Lyon 1 University The characterization of circulating tumor cells (CTCs) is a popular topic in translational research. This protocol describes a semi-automatic immunofluorescence (IF) assay for PD-L1 characterization and enumeration of CTCs in non-small cell lung cancer (NSCLC) patient samples. Medicine Measurements of Motor Function and Other Clinical Outcome Parameters in Ambulant Children with Duchenne Muscular Dystrophy Sara Nagy1,2, Simone Schmidt1,2, Patricia Hafner1,3, Andrea Klein1,4,5, Daniela Rubino-Nacht1, Vanya Gocheva1, Oliver Bieri6, Carole Vuillerot7, Ulrike Bonati1,2, Dirk Fischer1,2,3 1Division of Pediatric Neurology, University Children's Hospital Basel (UKBB), University of Basel, 2Department of Neurology, University Hospital Basel, University of Basel, 3Division of Neurology, University Clinic of Medicine, Cantonal Hospital Baselland, Bruderholz, 4Division of Pediatric Neurology, CHUV, 5Division of Neuropaediatrics, Inselspital, University Children's Hospital Bern, 6Department of Radiology, Division of Radiological Physics, University Hospital Basel, University of Basel, 7Hospices Civils de Lyon, Hôpital Femme-Mère-Enfant, L'Escale, Service de Médecine Physique et de Réadaptation Pédiatrique The aim of this study is to present the most reliable clinical outcome measures and their correlations with quantitative muscle MRI in ambulant patients with Duchenne muscular dystrophy. Neuroscience Modeling Charcot-Marie-Tooth Disease In Vitro by Transfecting Mouse Primary Motoneurons Arnaud Jacquier1,2, Valérie Risson1, Laurent Schaeffer1,3 1Institut NeuroMyoGène, CNRS UMR5310, INSERM U1217, Université Lyon, 2Unité Fonctionnelle de Neurogénétique Moléculaire, Hospices Civils de Lyon, 3Centre de Biotechnologie Cellulaire, Hospices Civils de Lyon The goal of this technique is to prepare a highly enriched culture of primary motoneurons (MNs) from murine spinal cord. To evaluate the consequences of mutations causing MN diseases, we describe here the isolation of these isolated MNs and their transfection by magnetofection. Medicine Confocal Time Lapse Imaging as an Efficient Method for the Cytocompatibility Evaluation of Dental Composites Ghania Nina Attik1, Kerstin Gritsch1,2, Pierre Colon1,3, Brigitte Grosgogeat1,2 1Laboratoire des Multimatériaux et Interfaces, UMR CNRS 5615, Université Lyon1, 2UFR d'Odontologie, Université Lyon1; Service de Consultations et de Traitements Dentaires, Hospices Civils de Lyon, 3UFR d'Odontologie, Université Paris Diderot; Service d'Odontologie, APHP, Hôpital Rothschild The most studied aspect of the biocompatibility of dental composites is their cytotoxicity 3D CLSM time lapse imaging combined with fluorescent signal quantification allows sensitive evaluation of the cell fate over time. Utilizing this method could be an efficient tool to assess the cytocompatibility of dental composites. Medicine Microarray-based Identification of Individual HERV Loci Expression: Application to Biomarker Discovery in Prostate Cancer Philippe Pérot1,2, Valérie Cheynet1,2, Myriam Decaussin-Petrucci1,3,4, Guy Oriol1,2, Nathalie Mugnier5, Claire Rodriguez-Lafrasse1,4,6, Alain Ruffion1,4,7, François Mallet1,2 1Joint Unit Hospices de Lyon-bioMérieux, 2Medical Diagnostic Discovery Department, BioMérieux, 3Department of Pathology and Cytology, Centre Hospitalier Lyon Sud, Hospices Civils de Lyon, 4Medical Faculty, Lyon 1 University, 5Data and Knowledge Laboratory, BioMérieux, 6Department of Biochemistry and Molecular Biology, Centre Hospitalier Lyon Sud, Hospices Civils de Lyon, 7Department of Urology, Centre Hospitalier Lyon Sud, Hospices Civils de Lyon Human endogenous retroviruses (HERV), which occupy 8% of the human genome, retain scarce coding capacities but a hundred thousand long terminal repeats (LTRs). A custom Affymetrix microarray was designed to identify individual HERV locus expression and was used on prostate cancer tissues as a proof of concept for future clinical studies.