Characterization of polymorphisms of genes ADH2, ADH3, ALDH2 and CYP2E1 and relationship to the alcoholism in a Colombian population

Objective: Identify and characterize polymorphisms of genes ADH2, ADH3, ALDH2 and CYP2E1 in a Colombian population residing in the city of Bogotá and determine its possible relationship to the alcoholism. Methods: ADH2, ADH3, ALDH2, and CYP2E1 genotypes a population of 148 individuals with non-problematic alcohol and 65 individuals with alcoholism were determined with TaqMan probes and PCR-RFLP. DNA was obtained from peripheral blood white cells. Results: Significant difference was found in family history of alcoholism and use of other psychoactive substances to compare alcoholics with controls. When allelic frequencies for each category (gender) were considered, frequency of A2 allele carriers in ADH2 was found higher in male patients than controls. In women, the relative frequency for c1 allele in CYP2E1 was lower in controls than alcoholics. The ALDH2 locus is monomorphic. No significant differences in allele distributions of the loci examined to compare two populations were observed, however when stratifying the same trend was found that these differences tended to be significant. Conclusions: This study allows us to conclude the positive association between family history of alcoholism and alcoholism suggesting that there is a favourable hereditary predisposition. Since substance dependence requires interaction of multiple genes, the combination of genotypes ADH2*2, CYP2E1*1 combined with genotype homozygous ALDH2*1 found in this study could be leading to the population to a potential risk to alcoholism.

Análise da expressão de APE1 após estresse oxidativo induzido em células proficientes e deficientes na via de reparo por excisão de nucleotídeos.

Riboflavin is a vitamin very important in aerobic organisms, as a precursor of many coenzymes involved in the electron transporter chain. However, after photosensitization of riboflavin with UV or visible light, it generates reactive oxygen species (ROS), which can oxidize the DNA. The repair of oxidative lesions on DNA occurs through the base excision repair pathway (BER), where APE1 endonuclease plays a central role. On the other hand, the nucleotide excision repair pathway (NER) repairs helix-distorting lesions. Recently, it was described the participation of NERproteins in the repair of oxidative damage and in stimulation of repair function fromAPE1. The aim of this research was to evaluate the cytotoxic effects of photosensitized riboflavin (RF*) in cells proficient and deficient in NER, correlating with APE1 expression. For this propose, the cells were treated with RF* and it was performed the cell viability assay, extraction of whole proteins, cells fractionation, immunoblotting, indirect immunofluorescence and analysis of polymorphisms of BER gens. The results evidenced that cells deficient in XPA and CSB proteins were more sensitive to RF*. However, XPC-deficient cells presented similar resistance to MRC5- SV cells, which is proficient in NER. These results indicate that XPA and CSB proteins have an important role on repair of oxidative lesions induced by RF*. Additionally, it was evidenced that single nucleotide polymorphisms (SNPs) in BER enzymes may influence in sensitivity of NER-deficient cell lines. Concerning the APE1 expression, the results showed that expression of this protein after treatment with RF* only changed in XPC-deficient cells. Though, it was observed that APE1 is recruited and is bound to chromatin in MRC5-SV and XPA cells after treatment with RF*. The results also showed the induction of DNA damage after treatment with RF*, through the analysis of-H2AX, since the treatment promoted an increase of endogenous levels of this phosphorylated protein, which acts signaling double strand-break on DNA. On the other hand, in XPC-deficient cells, regardless of resistance of RF*, the endogenous levels of APE1 are extremely reduced when compared with other cell lines and APE1 is not bound to chromatin after treatment with RF*. These results conclude that RF* was able to induce cell death in NERdeficient cells, where XPA and CSB cells were more sensitive when compared with MRC5-SV and XPC-deficient cells. This last result is potentially very interesting, since XPC-deficient cell line presents low levels of APE1. Additionally, the results evidenced that APE1 protein can be involved in the repair of oxidative damage induced by RF*, because APE1 is recruited and bound strongly to chromatin after treatment.

Molecular characterization of hairy fleabane using RAPD.

2016

Tratamento não-farmacológico do Diabetes Mellitus

O objetivo geral da presente monografia visa revisar as atualidades e condutas no tratamento não farmacológico do Diabetes Mellitus tipos 1 (DM1) e 2 (DM2), averiguando e estudando os benefícios ou malefícios do tratamento não farmacológico da presente comorbidade. Segundo estimativas da Organização Mundial de Saúde (OMS), o número de diabéticos no mundo em 2000 era de 177 milhões, com perspectiva de alcançar 350 milhões em 2025. Os estudos reportam os benefícios e os potenciais efeitos adversos da dieta alimentar associada a atividade física, entre outros. Esta revisão tem como prioridade revisar e reunir informações da literatura internacional, a respeito do tema proposto, divulgando as informações controversas e condutas estabelecidas de diferentes autores, evidenciando seus riscos e benefícios, contribuindo para o planejamento de outras pesquisas científicas. Foram analisados relevantes estudos publicados e as bases de dados: SciELO, PubMed, entre outros. A efetividade de uma ação em saúde está relacionada à eficácia da medida, à precisão diagnóstica, à aderência do médico, à aderência do paciente e ao nível de cobertura. Pode-se concluir que o DM afeta de maneira efetiva os ajustes fisiológicos relacionados ao metabolismo de carboidratos, trazendo consequências desastrosas para os demais sistemas fisiológicos, principalmente o vascular, resultando em doenças que podem em última instância, levar à morte. Estudos adicionais envolvendo exercício físico, diabetes mellitus ainda precisam ser realizados, com o intuito de analisar as respostas fisiológicas ao exercício físico e suas associações com polimorfismos genéticos específicos.