Improving the accuracy of recombinant protein production through integration of bioinformatics, statistical and mass spectrometry methodologies.

Heterologous protein production is a key technology for biotechnological, health sciences and many other research fields. Various approaches have been developed for its optimization, but the research emphasis has been on optimization of protein yield rather than protein quality. In this study, we have established a workflow for synthetic gene optimization for heterologous protein expression that combines bioinformatics, laboratory experiments, mass spectrometry and statistical analysis. Two gene primary structure analysis platforms, Anaconda and EuGene, and multivariate optimization methods were employed to re-design the Plasmodium falciparum lysyl-tRNA synthetase gene for optimal expression in Escherichia coli. Synthetic genes were expressed from common vectors, and amino acid mis-incorporations in the expressed proteins were detected and quantified using mass spectrometry. The association between the identified amino acid mis-incorporations and 23 gene variables was then analysed. The synthetic genes yielded significantly higher levels of protein relative to the wild-type gene, but 71 amino acid mis-incorporation sites were observed along the whole protein and across the synthetic genes that were statistically associated with specific codons and protein secondary structures. The optimization method that led to production of the most accurate protein was based on a multivariate approach that combined variables that are known to influence mRNA translation.

Estudo inicial sobre o desempenho de população com idade acima de 60 anos nos testes de stroop e rey / Initial study about the performance of elderly people under 60 years in stroop and rey tests

Estudos têm mostrado um aumento da população idosa em todo o mundo, sendo a expectativa média de vida do brasileiro, em 2025, de 80 anos. No entanto, observa-se uma carência de dados normativos de testes neuropsicológicos para a população de idosos brasileiros. Considerando a importância da caracterização de funções cognitivas como memória e atenção, este estudo teve como objetivo: 1) obter dados preliminares sobre o desempenho de idosos no teste de atenção de Stroop e no teste de memória verbal de Rey; 2)comparar o desempenho entre idosos com menos de 70 anos comaqueles mais velhos. Participaram do estudo 92 sujeitos, com idade entre 60 e 88 anos de ambos os sexos (47 mulheres e 45 homens). Resultados mostraram que idosos com mais de 70 anos apresentam desempenho inferior nas provas de atenção e memória. Idosos com maior escolaridade foram melhores em todas as etapas do Stroop eem metade das provas do Rey. Não foram observadas diferenças com relação ao sexo(AU)