Carlos III Health Institute 11 articles published in JoVE Medicine Description of a Swine Infant Model of Volume-Controlled Hemorrhagic Shock Alicia Rodríguez Martínez1,2,3, Sara de la Mata Navazo1,2,3, Gema Manrique Martín1,2,3, Sarah Nicole Fernández Lafever1,2,3, Laura Butragueño-Laiseca1,2,3, María Slöcker Barrio1,2,3, Rafael González Cortés1,2,3, Laura Herrera Castillo1,2,3, Santiago Mencía Bartolomé1,2,3, Jimena del Castillo Peral1,2,3, María José Solana García1,2,3, Débora Sanz Álvarez1,2,3, Raquel Cieza Asenjo1,2,3, Jorge López-González1,2,3, María José Santiago Lozano1,2,3, Daniel Moreno Leira1,2,3, Jesús López-Herce Cid1,2,3, Javier Urbano Villaescusa1,2,3 1Pediatric Intensive Care Department, Gregorio Marañón University General Hospital, 2Gregorio Marañón Health Research Institute, 3Primary Care Interventions to Prevent Maternal and Child Chronic Diseases of Perinatal and Development Origin Network (RICORS), Carlos III Health Institute This article aims to provide researchers with a detailed and accessible guide to set up an infant porcine model of hemorrhagic shock. Cancer Research In Vivo Inhibition of MicroRNA to Decrease Tumor Growth in Mice Julia Ramirez-Moya1,2, León Wert-Lamas1, Adrián Acuña-Ruiz1,2, Miguel A. Zaballos1,2, Pilar Santisteban1,2 1Instituto de Investigaciones Biomédicas, CSIC-UAM, 2Ciberonc, Instituto de Salud Carlos III This protocol describes xenograft and orthotopic mouse models of human thyroid tumorigenesis as a platform to test microRNA-based inhibitor treatments. This approach is ideal to study the function of non-coding RNAs and their potential as new therapeutic targets. Bioengineering Simultaneous Electrical and Mechanical Stimulation to Enhance Cells' Cardiomyogenic Potential Aida Llucià-Valldeperas1,2, Ramon Bragós3, Antoni Bayés-Genís1,4,5,6 1Insuficiencia Cardiaca y Regeneración Cardiaca (ICREC) Research Program, Health Science Research Institute Germans Trias i Pujol, 2Amsterdam Universitair Medisch Centrum (UMC), Vrije Universiteit Amsterdam, Pulmonology and Physiology, Amsterdam Cardiovascular Sciences, 3Electronic and Biomedical Instrumentation Group, Departament d'Enginyeria Electrònica, Universitat Politècnica de Catalunya, 4Cardiology Service, Germans Trias i Pujol University Hospital, 5Department of Medicine, Universitat Autònoma de Barcelona, 6Centro de Investigación Biomédica en Red (CIBER) Cardiovascular, Instituto de Salud Carlos III Here we present a protocol for training a cell population using electrical and mechanical stimuli emulating cardiac physiology. This electromechanical stimulation enhances the cardiomyogenic potential of the treated cells and is a promising strategy for further cell therapy, disease modeling, and drug screening. Engineering Effective Analysis of Human Exposure Conditions with Body-worn Dosimeters in the 2.4 GHz Band Silvia de Miguel-Bilbao1, Juan Blas2, Victoria Ramos1 1Telemedicine and e-Health Research Unit, Instituto de Salud Carlos III, 2Signal Theory and Communications, and Telematic Engineering Department, Universidad de Valladolid This study describes a protocol to measure exposure levels in the 2.4 GHz band, avoiding the uncertainties caused by the use of personal exposimeters as measuring devices. These alterations of the exposure levels should be taken into account, especially in compliance testing, where exposure limits are defined from non-perturbed data. Medicine An Experimental Model of Diet-Induced Metabolic Syndrome in Rabbit: Methodological Considerations, Development, and Assessment Óscar Julián Arias-Mutis1,2,3, Patricia Genovés1,2,3, Conrado J. Calvo1,2,4, Ana Díaz5, Germán Parra2,3, Luis Such-Miquel6, Luis Such2, Antonio Alberola2, Francisco Javier Chorro1,3, Manuel Zarzoso6 1CIBERCV, Instituto de Salud Carlos III, 2Department of Physiology, Universitat de València, 3INCLIVA, 4Department of Electronic Engineering, Universidad Politécnica de Valencia, 5UCIM, Universitat de València, 6Department of Physiotherapy, Universitat de València We describe methods to develop an experimental model of diet-induced metabolic syndrome (MetS) in rabbits using a high-fat, high-sucrose diet. Animals developed central obesity, mild hypertension, pre-diabetes, and dyslipidemia, thus reproducing the main components of human MetS. This chronic model will allow acquisition of knowledge underlying mechanisms of disease progression. Medicine Executing Complexity-Increasing Queries in Relational (MySQL) and NoSQL (MongoDB and EXist) Size-Growing ISO/EN 13606 Standardized EHR Databases Ricardo Sánchez-de-Madariaga1, Adolfo Muñoz1, Antonio L Castro1, Oscar Moreno1, Mario Pascual1 1Telemedicine and Information Society Department, Health Institute "Carlos III" This study compares relational and non-relational (NoSQL) standardized medical information systems. The computational complexity of the response times of querying such database management systems (DBMS) is computed using doubling-sized databases. These results help the discussion of the appropriateness of each database approach to different scenarios and problems. Immunology and Infection Screening Assays to Characterize Novel Endothelial Regulators Involved in the Inflammatory Response María Ángeles Higueras1, Lidia Jiménez-García1, Sandra Herranz1, Sonsoles Hortelano*1, Alfonso Luque*1 1Pharmacological Therapies Unit, Research Institute for Rare Diseases, Institute of Health Carlos III Vascular endothelium tightly controls leukocyte recruitment. Inadequate leukocyte extravasation contributes to human inflammatory diseases. Therefore, searching for novel regulatory elements of endothelial activation is necessary to design improved therapies for inflammatory disorders. Here, we describe a comprehensive methodology to characterize novel endothelial regulators that can modify leukocyte trafficking during inflammation. Immunology and Infection Functional Characterization of Regulatory Macrophages That Inhibit Graft-reactive Immunity Jordi Ochando1,2, Patricia Conde1,2 1Department of Medicine, Icahn School of Medicine at Mount Sinai, 2Immunología de Trasplantes, Centro Nacional de Microbiología, Instituto de Salud Carlos III Macrophages are plastic cells of the hematopoietic system that have a crucial role in protective immunity and homeostasis. In this report, we describe optimized in vitro techniques to phenotypically and functionally characterize graft-infiltrating regulatory macrophages that accumulate in the transplanted organ under tolerogenic conditions. Immunology and Infection Methodology for the Study of Horizontal Gene Transfer in Staphylococcus aureus Fabio Cafini*1,2, Nguyen Thi Le Thuy*3, Federico Román4, José Prieto5, Sarah Dubrac6,7, Tarek Msadek6,7, Kazuya Morikawa1 1Division of Biomedical Science, Faculty of Medicine, University of Tsukuba, 2Department of Basic Biomedical Science, Universidad Europea de Madrid, 3Human Biology Program, School of Integrative and Global Majors, University of Tsukuba, 4Laboratory of Nosocomial Infections, Department of Bacteriology, Centro Nacional de MicrobiologÍa, Instituto de Salud Carlos III, 5Division of Microbiology, Department of Medicine, School of Medicine, Universidad Complutense, 6Biology of Gram-Positive Pathogens, Department of Microbiology, Institut Pasteur, Paris, France, 7ERL3526, CNRS, Paris, France We describe here three different protocols for the in vitro investigation of conjugation, transduction, and natural transformation in Staphylococcus aureus. Biology A Comparative Study of Drug Delivery Methods Targeted to the Mouse Inner Ear: Bullostomy Versus Transtympanic Injection Silvia Murillo-Cuesta*1,2,3, Néstor Vallecillo*1, Rafael Cediel1,2,4, Adelaida M. Celaya1,2, Luis Lassaletta3,5, Isabel Varela-Nieto1,2,3, Julio Contreras1,2,4 1Instituto de Investigaciones Biomédicas (IIBm) Alberto Sols CSIC-UAM, 2Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III (ISCIII), 3Instituto de Investigación Sanitaria La Paz (IdiPAZ), 4Facultad de Veterinaria, Universidad Complutense de Madrid, 5Departmento de Otorrino laringología, Hospital Universitario La Paz Two microsurgery approaches for local drug delivery to the inner ear are described here and compared in terms of impact on hearing parameters, cochlear cytoarchitecture and expression of inflammatory markers. Immunology and Infection Phenotypic Characterization of Macrophages from Rat Kidney by Flow Cytometry Alfonso Rubio-Navarro1, Melania Guerrero-Hue1, Beatriz Martín-Fernandez2, Isabel Cortegano3, Elena Olivares-Alvaro2, Natalia de las Heras2, Mario Alía3, Belén de Andrés3, María Luisa Gaspar3, Jesús Egido1, Juan Antonio Moreno1 1Renal, Vascular and Diabetes Research Lab, IIS-Fundaciòn Jiménez Dìaz, Autonoma University, 2Department of Physiology, Faculty of Medicine, Complutense University, 3Department of Immunology, Centro Nacional de Microbiologìa, Instituto de Salud Carlos III (ISCIII) This manuscript describes a detailed protocol for phenotypic and quantitative analysis of resident macrophages from rat kidneys by flow cytometry. The resulting stained cells can be also used for other applications, including cell sorting, gene expression analysis or functional studies, thus increasing the information obtained in the experimental model.