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Universidade de São Paulo

15 ARTICLES PUBLISHED IN JoVE

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Environment

Mass Production of Genetically Modified Aedes aegypti for Field Releases in Brazil
Danilo O. Carvalho 1,2, Derric Nimmo 1, Neil Naish 1, Andrew R. McKemey 1, Pam Gray 1, André B. B. Wilke 3, Mauro T. Marrelli 3, Jair F. Virginio 4, Luke Alphey 1,5, Margareth L. Capurro 2,6
1Oxitec Ltd, 2Departamento de Parasitologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, 3Departamento de Epidemiologia, Universidade de São Paulo, 4Moscamed Brasil, 5Deptartment of Zoology, University of Oxford, 6Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular (INCT-EM)

To achieve population suppression of Aedes aegypti using the RIDL® (Release of Insects carrying a Dominant Lethal) system, large numbers of male mosquitoes need to be released. This requires the use of mass rearing techniques and technology to provide reliable systems to obtain the maximum number of high quality male mosquitoes.

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JoVE Core

Quantum State Engineering of Light with Continuous-wave Optical Parametric Oscillators
Olivier Morin 1, Jianli Liu 1, Kun Huang 1,2, Felippe Barbosa 3, Claude Fabre 1, Julien Laurat 1
1Laboratoire Kastler Brossel, Université Pierre et Marie Curie, Ecole Normale Supérieure, CNRS, 2State Key Laboratory of Precision Spectroscopy, East China Normal University, 3Instituto de Física, Universidade de São Paulo

We describe the reliable generation of non-Gaussian states of traveling optical fields, including single-photon states and coherent state superpositions, using a conditional preparation method operated on the non-classical light emitted by optical parametric oscillators. Type-I and type-II phase-matched oscillators are considered and common procedures, such as the required frequency filtering or the high-efficiency quantum state characterization by homodyning, are detailed.

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Biology

An Anoxia-starvation Model for Ischemia/Reperfusion in C. elegans
Bruno B. Queliconi 1, Alicia J. Kowaltowski 1, Keith Nehrke 2
1Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, 2Department of Medicine, Nephrology Division, University of Rochester Medical Center, School of Medicine and Dentistry

A protocol is described that uses anoxia/starvation in C. elegans to model ischemia/reperfusion. Functional outcomes include increased mortality, visible abnormalities in GFP-labeled neuronal processes, and impaired behavioral responses that require neuronal function.

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Neuroscience

RNA-Seq Analysis of Differential Gene Expression in Electroporated Chick Embryonic Spinal Cord
Felipe M. Vieceli 1, C.Y. Irene Yan 1
1Department of Cell and Developmental Biology, Universidade de São Paulo

This video shows the basic steps for performing whole transcriptome analysis on dissected chick embryonic spinal cord samples after transfection with in ovo electroporation.

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Immunology and Infection

Quantification of three DNA Lesions by Mass Spectrometry and Assessment of Their Levels in Tissues of Mice Exposed to Ambient Fine Particulate Matter
Tiago Franco de Oliveira *1,2, Antonio Anax Falcão de Oliveira *1, Miriam Lemos 3, Mariana Veras 3, Paulo Hilário Nascimento Saldiva 3,4, Marisa Helena Gennari de Medeiros 5, Paolo Di Mascio 5, Ana Paula de Melo Loureiro 1
1Departamento de Análises Clínicas e Toxicológicas, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, 2Departamento de Farmacociências, Universidade Federal de Ciências da Saúde de Porto Alegre, 3Laboratório de Poluição Atmosfêrica Experimental - LIM05, Hospital das Clínicas, Faculdade de Medicina, Universidade de São Paulo, 4Instituto de Estudos Avançados, Universidade de São Paulo, 5Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo

We describe here methods for sensitive and accurate quantification of the lesions 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo), 1,N6-etheno-2'-deoxyadenosine (1,N6-dAdo) and 1,N2-etheno-2'-deoxyguanosine (1,N2-dGuo) in DNA. The methods were applied to the assessment of the effects of ambient fine particulate matter (PM2.5) in tissues (lung, liver and kidney) of exposed A/J mice.

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Biochemistry

Assessment of Submitochondrial Protein Localization in Budding Yeast Saccharomyces cerevisiae
Fernando Gomes 1, Helena Turano 1, Angélica Ramos 1, Luis E. S. Netto 1
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.

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Study of Experimental Organ Donation Models for Lung Transplantation

Study of Experimental Organ Donation Models for Lung Transplantation
Natalia Aparecida Nepomuceno 1, Liliane Moreira Ruiz 1, Paolo Oliveira-Melo 1, Náthaly Christinie Ikeoka Eroles 1, Isabela Gomes Viana 1, Paulo Manuel Pêgo-Fernandes 1, Karina Andrighetti de Oliveira Braga 1
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.

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Cancer Research

A Reporter Assay to Analyze Intronic microRNA Maturation in Mammalian Cells
Guilherme Henrique Gatti da Silva 1, Patricia P. Coltri 1
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.

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Pre-RNA Splicing Analysis: From Basic Science To Applied Research
Patricia Pereira Coltri 1
1Departamento de Biologia Celular e do Desenvolvimento, Instituto de Ciências Biomédicas, Universidade de São Paulo

Pre-RNA Splicing Analysis: From Basic Science To Applied Research

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Multitargeted Analyses are Instrumental for Microbial Ecology Studies
Otávio Guilherme Gonçalves de Almeida 1, Elaine Cristina Pereira De Martinis 1
1Departamento de Análises Clínicas, Toxicológicas e Bromatológicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo

Multitargeted Analyses are Instrumental for Microbial Ecology Studies

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JoVE Journal

Ex Vivo Porcine Experimental Model for Studying and Teaching Lung Mechanics
Milena Gomes Parzianello Egúsquiza 1, Denise Aya Otsuki 1, José Otávio Costa Auler Junior 1
1Faculdade de Medicina da, Universidade de São Paulo

We present an ex vivo pig lung model for the demonstration of pulmonary mechanics and alveolar recruitment maneuvers for teaching purposes. The lungs can be used for more than one day (up to five days) with minimal changes in pulmonary mechanics variables.

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Biology

Process Development for the Spray-Drying of Probiotic Bacteria and Evaluation of the Product Quality
Letícia Kakuda 1, Yeily Jaramillo 1, Fabian C. Niño-Arias 1, Manuella F. de Souza 1, Edemilson C. Conceição 2, Virgínia F. Alves 2, Otávio G.G. de Almeida 1, Elaine C. P. De Martinis 1, Wanderley P. Oliveira 1
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.

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Biology

Profiling the Bacterial Community of Fermenting Traminette Grapes during Wine Production using Metagenomic Amplicon Sequencing
Adrienne Goppold 1, Louis Conradie 1, Otávio Guilherme Gonçalves de Almeida 2, Elaine Cristina Pereira De Martinis 2, Folarin Anthony Oguntoyinbo 1
1A.R. Smith Department of Chemistry and Fermentation Sciences, Appalachian State University, 2Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo

Here, we present a protocol to describe amplicon metagenomic for determining the bacterial community of Traminette grapes, fermenting grapes, and final wine.

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Immunology and Infection

Demystifying In Vivo Bioluminescence Imaging of a Chagas Disease Mouse Model for Drug Efficacy Studies
Adriana C. da Silva 1, Jadel M. Kratz 2, Priscylla G. M. Morgado 1,3, Lucio H. Freitas-Junior 1, Carolina B. Moraes 1,4
1Departamento de Microbiologia, Instituto de Ciências Biomédicas (ICB), Universidade de São Paulo, 2Drugs for Neglected Diseases initiative (DNDi) Latin America, 3Programa de Pós-Graduação em Microbiologia e Imunologia, Universidade Federal de São Paulo, 4Departamento de Análises Clínicas e Toxicológicas, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo

The protocol provided here describes the detailed steps to perform drug efficacy studies in a bioluminescent model of Trypanosoma cruzi infection, with a focus on data acquisition, analysis, and interpretation. Troubleshooting and quality control procedures to minimize technical issues are also provided.

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Embryo Transfer Surgery via Laparotomy in Gilts

Embryo Transfer Surgery via Laparotomy in Gilts
Michelle Silva Araujo 1, André Pegoraro Poor 1, Valéria Vieira Chida 1, Gustavo Torres 1, Ligiane Leme 1, Nicolle de S. B. Santos 1, Ângela May Suzuki 1, Tulio T. Yoshinaga 1, Giovanna C. Tolezano 1, André Costa 1, Evelyn L. Ribeiro 1, Taina da S. Bicalho 1, Deibs Barbosa 1, Karina A. de O. Braga 2, Luciano Abreu Brito 1, Luiz C. de C. Júnior 1, Mayana Zatz 1, Silvano M. A. Raia 1,3, Ernesto Goulart 1
1Centro de Estudos sobre o Genoma Humano e Células-Tronco, Instituto de Biociências, Universidade de São Paulo, 2Instituto do Coração (Incor), Hospital das Clínicas HCFMUSP, Faculdade de Medicina, Universidade de São Paulo, 3Faculdade de Medicina, Universidade de São Paulo

This protocol describes the application of the surgical technique used to transfer cloned pig embryos via laparotomy in gilts.

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