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Instituto de Salud Carlos III

12 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Functional Characterization of Regulatory Macrophages That Inhibit Graft-reactive Immunity
Jordi Ochando 1,2, Patricia Conde 1,2
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.

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Immunology and Infection

Methodology for the Study of Horizontal Gene Transfer in Staphylococcus aureus
Fabio Cafini *1,2, Nguyen Thi Le Thuy *3, Federico Román 4, José Prieto 5, Sarah Dubrac 6,7, Tarek Msadek 6,7, Kazuya Morikawa 1
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.

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Engineering

Effective Analysis of Human Exposure Conditions with Body-worn Dosimeters in the 2.4 GHz Band
Silvia de Miguel-Bilbao 1, Juan Blas 2, Victoria Ramos 1
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.

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Medicine

An Experimental Model of Diet-Induced Metabolic Syndrome in Rabbit: Methodological Considerations, Development, and Assessment
Óscar Julián Arias-Mutis 1,2,3, Patricia Genovés 1,2,3, Conrado J. Calvo 1,2,4, Ana Díaz 5, Germán Parra 2,3, Luis Such-Miquel 6, Luis Such 2, Antonio Alberola 2, Francisco Javier Chorro 1,3, Manuel Zarzoso 6
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.

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JoVE Core

Simultaneous Electrical and Mechanical Stimulation to Enhance Cells' Cardiomyogenic Potential
Aida Llucià-Valldeperas 1,2, Ramon Bragós 3, Antoni Bayés-Genís 1,4,5,6
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.

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Cancer Research

In Vivo Inhibition of MicroRNA to Decrease Tumor Growth in Mice
Julia Ramirez-Moya 1,2, León Wert-Lamas 1, Adrián Acuña-Ruiz 1,2, Miguel A. Zaballos 1,2, Pilar Santisteban 1,2
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.

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Medicine

Mobile Device-assisted Dietary Ecological Momentary Assessments for the Evaluation of the Adherence to the Mediterranean Diet in a Continuous Manner
Anna Boronat 1, Joana Clivillé-Pérez 2, Natalia Soldevila-Domenech 1,2, Laura Forcano 1,3, Nieves Pizarro 1,2, Montserrat Fitó 3,4, Helmut Schröder 4,5, Karine Fauria 6, Rafael de la Torre 1,2,3
1Integrative Pharmacology and Systems Neurosciences Research Group, Neurosciences Research Program, Hospital del Mar Medical Research Institute (IMIM), 2Department of Experimental and Health Sciences, University Pompeu Fabra, 3CIBER de Fisiopatología de la Obesidad y la Nutrición (CIBEROBN), Instituto de Salud Carlos III, 4Cardiovascular Risk and Nutrition Research Group, Hospital del Mar Medical Research Institute (IMIM), 5CIBER de Epidemiología y Salud Pública (CIBERESP), Instituto de Salud Carlos III (ISCIII), 6Barcelonaβeta Brain Research Center (BBRC), Pasqual Maragall Foundation

Presented here is a study protocol aimed at monitoring continuous adherence to the Mediterranean diet (MedDiet) by means of ecological momentary assessments. The method evaluates the intake of key food groups of the MedDiet and calculates an index of adherence.

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Medicine

Myocardial Infarction by Percutaneous Embolization Coil Deployment in a Swine Model
Daina Martínez-Falguera *1,2, Edgar Fadeuilhe *3, Albert Teis 3, Julia Aranyo 3, Raquel Adeliño 1,3, Felipe Bisbal 3,4, Oriol Rodriguez-Leor 3,4, Carolina Gálvez-Montón 1,3,4
1ICREC Research Program, Germans Trias i Pujol Health Research Institute (IGTP), 2Faculty of Medicine, University of Barcelona (UB), 3Heart Institute (iCOR), Germans Trias i Pujol University Hospital, 4CIBER Cardiovascular, Instituto de Salud Carlos III

Myocardial infarction (MI) animal models that emulate the natural process of the disease in humans are crucial to understanding pathophysiological mechanisms and testing the safety and efficacy of new emergent therapies. Here, we describe an MI swine model created by deploying a percutaneous embolization coil.

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Biology

Isolation of Mitochondria from Mouse Skeletal Muscle for Respirometric Assays
Juan Diego Hernández-Camacho 1,2, Cristina Vicente-García 1, Ana Sánchez-Cuesta 1,2, Daniel J. M. Fernandez-Ayala 1,2, Jaime J. Carvajal 1, Plácido Navas 1,2
1Centro Andaluz de Biología del Desarrollo, CSIC-UPO-JA, Universidad Pablo de Olavide, 2CIBERER, Instituto de Salud Carlos III

Here, we describe a detailed method for mitochondria isolation from mouse skeletal muscle and the subsequent analysis of respiration by Oxygen Consumption Rate (OCR) using microplate-based respirometric assays. This pipeline can be applied to study the effects of multiple environmental or genetic interventions on mitochondrial metabolism.

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Neuroscience

Enrichment of Astrocyte-Derived Extracellular Vesicles from Human Plasma
Natalia Valle-Tamayo 1,2, Rocío Pérez-González 2,3, Gemma Chiva-Blanch 4,5, Olivia Belbin 1,2, Sara Serrano-Requena 1,2, Sònia Sirisi 1,2, Alba Cervantes González 1,2, Oriol Giró 4,5, Érika Sánchez-Aced 1,2, Oriol Dols-Icardo 1,2, Daniel Alcolea 1,2, María Carmona-Iragui 1,2, Amanda Jimenez 4,5, Alberto Lleó 1,2, Juan Fortea 1,2, M. Florencia Iulita 1,2
1Sant Pau Memory Unit, Department of Neurology, Hospital de la Santa Creu i Sant Pau, Biomedical Research Institute Sant Pau, Universitat Autònoma de Barcelona, 2Center of Biomedical Investigation Network for Neurodegenerative Diseases (CIBERNED), 3Movement Disorders Unit, Department of Neurology, Hospital de la Santa Creu i Sant Pau, 4Department of Endocrinology and Nutrition, Hospital Clínic de Barcelona, August Pi i Sunyer Biomedical Research Institute - IDIBAPS, 5Spanish Biomedical Research Network in Physiopathology of Obesity and Nutrition (CIBEROBN)

This protocol describes the enrichment of astrocyte-derived extracellular vesicles (ADEVs) from human plasma. It is based on the separation of EVs by polymer precipitation, followed by ACSA-1 based immunocapture of ADEVs. Analysis of ADEVs may offer clues to study changes in inflammatory pathways of living patients, non-invasively by liquid biopsy.

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Methods of Isolation and Purification of Extracellular Vesicles from Different Biological Matrixes: Special Issue at a Glance
Gemma Chiva-Blanch 1,2
1Department of Endocrinology and Nutrition August Pi i Sunyer Biomedical Research Institute - IDIBAPS, Hospital Clínic of Barcelona, 2Biomedical Network Research Centre on Obesity and Nutrition Physiopathology (CIBEROBN), Instituto de Salud Carlos III

Methods of Isolation and Purification of Extracellular Vesicles from Different Biological Matrixes: Special Issue at a Glance

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Biology

Quantifying Fitness Costs in Transgenic Aedes aegypti Mosquitoes
Adeline E. Williams 1,2, Irma Sanchez-Vargas 1, Lindsay E. Martin 2,3, Ines Martin-Martin 2,4, Susi Bennett 1, Ken E. Olson 1, Eric Calvo 2
1Center for Vector-borne Infectious Diseases, Department of Microbiology, Immunology, and Pathology, Colorado State University, 2Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 3Department of Biological Sciences, Vanderbilt University, 4National Center for Microbiology, Instituto de Salud Carlos III

The present protocol describes how to measure common life parameter data in Aedes aegypti mosquitoes, including fecundity, wing size, fertility, sex ratio, viability, development times, male contribution, and adult longevity. These measurements can be used to assess the fitness of transgenic mosquitoes.

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