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5 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Parasite Induced Genetically Driven Autoimmune Chagas Heart Disease in the Chicken Model
Antonio R. L. Teixeira 1, Nadjar Nitz 1, Francisco M. Bernal 1, Mariana M. Hecht 1
1Chagas Disease Multidisciplinary Research Laboratory, University of Brasilia

The inoculation of Trypanosoma cruzi in fertile eggs prior to incubation renders the parasite kDNA minicircle integration in embryo cells genome. Crossbreeding reveals the vertical transfer of the mutations to progeny. The kDNA integrates into coding regions at several chromosomes and the chickens die with an inflammatory autoimmune heart disease.

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Chemistry

CN-GELFrEE - Clear Native Gel-eluted Liquid Fraction Entrapment Electrophoresis
Rafael D. Melani *1,2, Henrique S. Seckler *1, Owen S. Skinner 1, Luis H. F. Do Vale 1,3, Adam D. Catherman 1, Pierre C. Havugimana 1, Marcelo Valle de Sousa 3, Gilberto B. Domont 2, Neil L. Kelleher 1, Philip D. Compton 1
1Departments of Chemistry and Molecular Biosciences, Chemistry of Life Processes Institute, Proteomics Center of Excellence, Robert H. Lurie Comprehensive Cancer Center, Northwestern University, 2Institute of Chemistry, Proteomics Unit, Federal University of Rio de Janeiro, 3Department of Cell Biology, Brazilian Center for Protein Research, Laboratory of Biochemistry and Protein Chemistry, University of Brasilia

This protocol describes how to prepare and perform clear native gel-eluted liquid fraction entrapment electrophoresis (CN-GELFrEE), a native separation technique for non-covalent biomolecular assemblies and proteins from heterogeneous samples that is compatible with various downstream protein analysis techniques.

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Immunology and Infection

Sexual Transmission of American Trypanosomes from Males and Females to Naive Mates
Adriana B. Almeida 1, Perla F. Araújo 1, Francisco M. Bernal 1, Ana de Cassia Rosa 1, Sebastião A. Valente 2, Antonio R.L. Teixeira 1
1Chagas Disease Multidisciplinary Research Laboratory, Faculty of Medicine, University of Brasilia, 2Instituto Evandro Chagas

The Trypanosoma cruzi agent of Chagas disease produces long-lasting asymptomatic infections that abruptly develop into clinically recognized pathology. The following research protocol describes a short-run family-based epidemiological study to unravel the T. cruzi infection transmitted sexually from parent to progeny.

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Immunology and Infection

A Set of Screening Techniques for a Quick Overview of the Neutrophil Function
Isabelle Souza Luz 1, Raquel Takaya 1, Daiane Gonzaga Ribeiro 1, Natanael Sales Silva 1, Lucas Fontes 2, Mariana S. Castro 1, Wagner Fontes 1
1Laboratory of Protein Chemistry and Biochemistry, Department of Cell Biology, University of Brasilia, 2Institute of Physics, University of Brasilia

This protocol features a set of neutrophil functional assays to be used as a screening method to cover functions from different signaling pathways. The protocol includes an initial and simple evaluation of cell viability, purity, reactive oxygen species production, real-time migration, phagocytosis, and a preliminary suggestion of neutrophil extracellular traps.

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Biochemistry

RGBradford: Protein Quantitation with a Smartphone Camera
Daniel C. Moreira 1
1Research Center in Morphology and Applied Immunology, Faculty of Medicine, University of Brasilia

This paper provides a protocol for protein quantification using the Bradford assay and a smartphone as an analytical device. Protein levels in samples can be quantified using color data extracted from a picture of a microplate taken with a smartphone.

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