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Evaluation of studies related to molecular, genetic and biochemical markers in aging and chronic degenerative diseases
Brazil (IFMSA-Brazil) - Faculdade de Medicina de Sao Jose do Rio Preto, Sao Jose do Rio Preto
Departament of Molecular Biology – Laboratory of Biochemistry and Molecular Biology Av. Brg. Faria Lima, 5416 - Vila Sao Pedro, São José do Rio Preto - SP, 15090-000
Profª. Dra. Doroteia Souza
Profª. Dra. Doroteia Souza
English, Portuguese, Spanish and Italian
Type of Research Project
- Basic science
What is the background of the project?
The research center in Biochemistry and Molecular Biology (NPBIM) focuses on the study of degenerative diseases, involving genetic factors (polymorphisms and gene expression) and biochemistry (lipids, proteins and markers of oxidative stress). This study works in the development of techniques in the identification of biomarkers, including genetic polymorphisms and gene expression related to angiogenesis, inflammation, oxidative stress and lipid metabolism may contribute to the clarification of the pathophysiology, diagnosis and therapeutics of cardiovascular, hepatic, neurological and cancer.
What is the aim of the project?
To evaluate the association of genetic and biochemical profiles involving pathways related to angiogenesis, oxidative stress, inflammation and lipid metabolism in patients with cardiovascular diseases, hepatopathies and neuropsychiatric disorders and develop new techniques to identify these processes.
What techniques and methods are used?
Extraction of genomic DNA from peripheral blood leukocytes (5 ml) collected with EDTA will be performed. The extraction technique consisted of the salting-out method (Salazar, 1998), performed in three stages: 1) lysis of blood cells; 2) deproteinization; 3) Deoxyribonucleic acid (DNA) precipitation and resuspension. Extraction of total ribonucleic acid (RNA) by Trizol (5 ml of peripheral blood) will also be performed according to the manufacturer's manual (Invitrogen-Thermo Fisher Scientific). Blood samples will be collected in tube with anticoagulant followed by centrifugation and extraction of total RNA. Single-phase solution containing phenol and guanidine isothiocyanate (TRIzol™ LS Reagent, Invitrogen) will be used according to the manufacturer's protocol. The concentration and purity of the extracted RNA will be evaluated in the NanoDrop equipment (NanoDrop Technologies, Inc., Wilmington, DE). An amount of RNA above 50ng / gL and a purity of between 1.7 and 1.9 is considered appropriate. In addition, after the extraction of genomic DNA, real-time polymerase chain reaction (PCR) polymorphism analysis will be done, with final solution volume of 5 samples in each well of the 96-well plate. The final solution in each well will be composed of the mixture of 1,375 III, Depc Water (Diethyl Pyrocarbonate), 2,5µL Master Mix Universal (Thermo ScientificTM), 0.125µL of the TaqMan®SNP Genotyping Assays Probe for each polymorphism and 1µL of diluted DNA. This protocol was indicated for standardization by the supplier company (Thermo Fisher Scientific).
What is the role of the student?
- The student will mainly observe
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The student should mainly follow and eventually participate in the processing of biological material and techniques for analysis of genetic polymorphisms and gene expression. Main activities: monitoring of biological samples collection; DNA extraction by saline precipitation; analysis of polymorphisms by real-time polymerase chain reaction (PCR); quantification of gene expression; tissue RNA extraction by Trizol®; introduction to the Laboratory's research lines and respective projects in progress.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
Yes, the laboratory promotes periodic scientific meetings, in addition, the student can participate in lectures, seminars and events promoted by the institution.
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a poster
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Basic laboratory and basic biochemical knowledge. Subjects passed: Molecular Biology
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Pre-Medical students from the American-British system - Students in biomedical fields
- Genetic variants related to angiogenesis and apoptosis in patients with glioma. Arq Neuropsiquiatr. 2018 Jun;76(6):393 -398. doi: 10.1590/0004-282X20180051. Calastri MCJ; Rodrigues NLTO; Hatori G; Gregório ML; Brancati CIFO; Zanovelo EM; Ferraz Filho JRL; Neiva CM; Rodrigues Junior ACP; Godoy MF; Lancelloti CLP; Tognola WA; Souza DRS.
- VEGF Polymorphisms Related to Higher Serum Leveis of Protein Identify Patients with Hepatocellular Carcinoma.Can J Gastroenterol Hepatol. 2016;2016:9607054. Epub 2016 Aug 31. Baitello ME; Tenani GD; Ferreira RF; Nogueira V; Pinhel MA; da Silva RC; da Silva RF; Fucuta PD; de Godoy MF1; Souza DR.
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