Universidade de Sao Paulo 7 articles published in JoVE Medicine Study of Experimental Organ Donation Models for Lung Transplantation Natalia Aparecida Nepomuceno1, Liliane Moreira Ruiz1, Paolo Oliveira-Melo1, Náthaly Christinie Ikeoka Eroles1, Isabela Gomes Viana1, Paulo Manuel Pêgo-Fernandes1, Karina Andrighetti de Oliveira Braga1 1Laboratório de Pesquisa em Cirurgia Torácica, Departamento de Cardiopneumologia, Instituto do Coracao, Faculdade de Medicina HCFMUSP, Universidade de São Paulo The present study shows the establishment of three different lung donation models (post-brain death donation, post-circulatory death donation, and post-hemorrhagic shock donation). It compares the inflammatory processes and pathological disorders associated with these events. Medicine Human Saphenous Vein Endothelial Cell Isolation and Exposure to Controlled Levels of Shear Stress and Stretch Thaís Girão-Silva*1,2, Miriam Helena Fonseca-Alaniz*1, Luís Alberto Oliveira Dallan1, Iuri Cordeiro Valãdao1, Gustavo Henrique Oliveira da Rocha1, José Eduardo Krieger1, Ayumi Aurea Miyakawa1 1Instituto do Coraçao (InCor), Hospital das Clinicas HCFMUSP, Faculdade de Medicina, Universidade de Sao Paulo, 2Cardiology Research Group, Faculty VI Medicine and Health Sciences, Carl von Ossietzky University of Oldenburg We describe a protocol to isolate and culture human saphenous vein endothelial cells (hSVECs). We also provide detailed methods to produce shear stress and stretch to study mechanical stress in hSVECs. Biology Process Development for the Spray-Drying of Probiotic Bacteria and Evaluation of the Product Quality Letícia Kakuda1, Yeily Jaramillo1, Fabian C. Niño-Arias1, Manuella F. de Souza1, Edemilson C. Conceição2, Virgínia F. Alves2, Otávio G.G. de Almeida1, Elaine C. P. De Martinis1, Wanderley P. Oliveira1 1Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, 2Faculdade de Farmácia, Universidade Federal De Goiás This protocol details the steps involved in the production and physicochemical characterization of a spray-dried probiotic product. Neuroscience Hypothalamic Kisspeptin Neurons as a Target for Whole-Cell Patch-Clamp Recordings Josiane do Nascimento Silva1, Thais T. Zampieri1, Henrique R. Vieira1, Renata Frazao1 1Department of Anatomy, Instituto de Ciencias Biomedicas, Universidade de Sao Paulo Here, we present a protocol to perform a whole-cell patch-clamp on brain slices containing kisspeptin neurons, the primary modulator of gonadotrophin-releasing hormone (GnRH) cells. By adding knowledge about kisspeptin neuron activity, this electrophysiological tool has served as the basis for significant advancements in the neuroendocrinology field over the last 20 years. Cancer Research A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells Guilherme Henrique Gatti da Silva1, Patricia P. Coltri1 1Departamento de Biologia Celular e do Desenvolvimento, Instituto de Ciências Biomédicas, Universidade de São Paulo We developed an intronic microRNA biogenesis reporter assay to be used in cells in vitro with four plasmids: one with intronic miRNA, one with the target, one to overexpress a regulatory protein, and one for Renilla luciferase. The miRNA was processed and could control luciferase expression by binding to the target sequence. Biochemistry Assessment of Submitochondrial Protein Localization in Budding Yeast Saccharomyces cerevisiae Fernando Gomes1, Helena Turano1, Angélica Ramos1, Luis E. S. Netto1 1Departamento de Genética e Biologia Evolutiva, Instituto de Biociências, Universidade de São Paulo Despite recent advances, many yeast mitochondrial proteins still remain with their functions completely unknown. This protocol provides a simple and reliable method to determine the submitochondrial localization of proteins, which has been fundamental for the elucidation of their molecular functions. Neuroscience Positron Emission Tomography Imaging for In Vivo Measuring of Myelin Content in the Lysolecithin Rat Model of Multiple Sclerosis Daniele de Paula Faria1, Caroline Cristiano Real1, Larissa Estessi de Souza1, Alexandre Teles Garcez2, Fabio Luis Navarro Marques1, Carlos Alberto Buchpiguel1 1Laboratory of Nuclear Medicine (LIM-43), Departamento de Radiologia e Oncologia, Faculdade de Medicina, Universidade de Sao Paulo, 2Nuclear Medicine Division, Hospital Sirio-Libanes This protocol has the aim of monitoring in vivo myelin changes (demyelination and remyelination) by positron emission tomography (PET) imaging in an animal model of multiple sclerosis.
