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Evaluation of Mitochondrial DNA polymorphisms in obese patients responders or not to clinical

Grant number: 14/17097-0
Support type:Regular Research Grants
Duration: December 01, 2014 - November 30, 2016
Field of knowledge:Health Sciences - Medicine
Principal Investigator:João Eduardo Nunes Salles
Grantee:João Eduardo Nunes Salles
Home Institution: Faculdade de Ciências Médicas da Santa Casa de São Paulo (FCMSCSP). Fundação Arnaldo Vieira de Carvalho. São Paulo , SP, Brazil
Assoc. researchers:Carlos Alberto Longui ; Nilza Maria Scalissi

Abstract

Obesity is increasing worldwide at a rate of epidemic. The etiology of obesity is complex, multifactorial, resulting from the interaction of genes, environment, lifestyle and emotional factors.The cellular growth and energy balance are related to mitochondrial metabolism efficiency. Therefore, genetic variants of mitochondrial DNA (mtDNA) may be involved in individual energy expenditure, having direct relation to weight. The mtDNA sequence analysis or PCR amplification techniques using tools can be used to detect these variants, allowing correlation between mtDNA alterations, energy expenditure and body mass index (BMI).In the obese population, we will evaluate the polymorphisms of mtDNA and we will compare with those with normal BMI.For this, we will select 50 obese who do regular monitoring in the outpatient Obesity, shopping with a control group of 50 individuals. Of each study subject a sample of 5ml of peripheral blood will be collected.For the identification of polymorphisms in mitochondrial DNA resequencing system based on the PCR for identifying variations in the hypervariable region of the human mitochondrial genome is used. The method consists of 5 steps using kit mitoSEQr " Resequencing System for the Human Mitochondrial Genome, Applied Biosystems ": 1) Obtaining DNA (genomic and mitochondrial) from linfomonocitárias peripheral blood cells. 2) Determine the concentration of DNA extracted by reading in a spectrophotometer at a wavelength of A260. 3) Reaction amplification of sequences of the PCR DNAM. 4) Purification of Sequencing Reactions. 5) Electrophoresis. (AU)