Extraction of hemoglobin with calixarenes and biocatalysis in organic media of the complex with pseudoactivity of peroxidase

The present work involves the use of p-tert-butylcalix[4,6,8]arene carboxylic acid derivatives ((t)Butyl[4,6,8]CH2COOH) for selective extraction of hemoglobin. All three calixarenes extracted hemoglobin into the organic phase, exhibiting extraction parameters higher than 0.90. Evaluation of the solvent accessible positively charged amino acid side chains of hemoglobin (PDB entry 1XZ2) revealed that there are 8 arginine, 44 lysine and 30 histidine residues on the protein surface which may be involved in the interactions with the calixarene molecules. The hemoglobin-(t)Butyl[6]CH2COOH complex had pseudoperoxidase activity which catalysed the oxidation of syringaldazine in the presence of hydrogen peroxide in organic medium containing chloroform. The effect of pH, protein and substrate concentrations on biocatalysis was investigated using the hemoglobin-(t)Butyl[6]CH2COOH complex. This complex exhibited the highest specific activity of 9.92 x 10(-2) U mg protein(-1) at an initial pH of 7.5 in organic medium. Apparent kinetic parameters (V'(max), K'(m), k'(cat) and k'(cat)/K'(m)) for the pseudoperoxidase activity were determined in organic media for different pH values from a Michaelis-Menten plot. Furthermore, the stability of the protein-calixarene complex was investigated for different initial pH values and half-life (t(1/2)) values were obtained in the range of 1.96 and 2.64 days. Hemoglobin-calixarene complex present in organic medium was recovered in fresh aqueous solutions at alkaline pH, with a recovery of pseudoperoxidase activity of over 100%. These results strongly suggest that the use of calixarene derivatives is an alternative technique for protein extraction and solubilisation in organic media for biocatalysis.

Differential expression of CDC25 phosphatases splice variants in human breast cancer cells

Background: CDC25 phosphatases control cell cycle progression by activating cyclin dependent kinases. The three CDC25 isoforms encoding genes are submitted to alternative splicing events which generate at least two variants for CDC25A and five for both CDC25B and CDC25C. An over-expression of CDC25 was reported in several types of cancer, including breast cancer, and is often associated with a poor prognosis. Nevertheless, most of the previous studies did not address the expression of CDC25 splice variants. Here, we evaluated CDC25 spliced transcripts expression in anti-cancerous drug-sensitive and resistant breast cancer cell lines in order to identify potential breast cancer biomarkers. Methods: CDC25 splice variants mRNA levels were evaluated by semi-quantitative RT-PCR and by an original real-time RT-PCR assay. Results: CDC25 spliced transcripts are differentially expres-sed in the breast cancer cell lines studied. An up-regulation of CDC25A2 variant and an increase of the CDC25C5/C1 ratio are associated to the multidrug-resistance in VCREMS and DOXOR breast cancer cells, compared to their sensitive counterpart cell line MCF-7. Additionally, CDC25B2 tran-script is exclusively over-expressed in VCREMS resistant cells and could therefore be involved in the development of certain type of drug resistance. Conclusions: CDC25 splice variants could represent interesting potential breast cancer prognostic biomarkers.

Effect of deworming on hemoglobin concentration in children from 2 to 15 years from the Bengo Province, Angola

The most common causes of anemia are micronutrient deficiencies, but other factors may influence namely inflammation, parasitic infections and inherited disorders. One strategy to combat micronutrient deficiencies is supplementation, yet, in zones with high prevalence of Schistosomiasis or Soil Transmitted Helminthes (STH), supplementation could be not sufficient. The aim of this study was to evaluate the effects of deworming, on hemoglobin concentration, in children from 2 to 15 years, from Bengo.

Ressonância magnética funcional: mapeamento do córtex motor através da técnica BOLD

A ressonância magnética funcional (RMf) é hoje uma ferramenta fundamental na investigação funcional do cérebro humano, quer em indivíduos saudáveis quer em pacientes com patologias diversas. É uma técnica complexa que necessita de uma aplicação cuidada e rigorosa, e uma compreensão dos mecanismos biofísicos a ela subjacentes, de modo a serem obtidos resultados fiáveis e com melhor aceitação clínica. O efeito BOLD (Blood Oxygenation Level Dependent) é o método mais utilizado para medir e estudar a actividade cerebral e baseia-se nas alterações das propriedades magnéticas da molécula hemoglobina. Com este Projecto propomo-nos optimizar um protocolo de RMf realizada com o efeito BOLD, em voluntários saudáveis, de modo a que este possa ser aplicado em futuros estudos de pacientes com patologias. ABSTRACT - Nowadays functional magnetic resonance imaging (fMRI) is a fundamental tool for the research of human brain function of healthy subjects or patients with several pathologies. It is a complex technique that requires a careful and rigorous application, and an understanding of its biophysical mechanisms, so that reliable results can be obtained with better clinical acceptance. The BOLD effect (Blood Oxygenation Level Dependent) is the most widely used method to measure and study the brain activity and its based on changes in magnetic properties of the hemoglobin molecule. The aim of this project was to optimize a BOLD fMRI protocol on healthy subjects, so it can be applied in future studies of patients with pathologies.

Processus interactionnel d’appropriation de la syntaxe et de variantes énonciatives diversifiées, indispensable pour l’accès à l’écrit

A investigação tem posto em evidência a correlação entre desenvolvimento linguístico e aprendizagem da leitura e da escrita. Procuramos, em primeiro lugar, explicar, do ponto de vista da linguística, as relações entre oral e escrita focalizando a linguagem da criança desde as primeiras produções à produção autónoma de variantes sintacticamente diversificadas. De seguida, abordamos a questão da passagem do oral à escrita dando especial atenção ao impacto sobre o funcionamento cognitivo do trabalho de ditado ao adulto. Finalmente, propomos pistas de reflexão sobre modalidades de interacção necessárias à entrada no modo escrito e ao seu percurso de aprendizagem dando conta de resultados de investigação acção e formação com a abordagem «Coup de pouce Langage».

Localização ótima de apoios em estruturas do tipo viga utilizando o método dos algoritmos genéticos

Dissertação de Mestrado para obtenção do grau de Mestre em Engenharia Mecânica Ramo de Manutenção e Produção

Optimização do posicionamento de câmaras de vídeo-vigilância

Dissertação de Mestrado em Engenharia de Redes de Comunicação e Multimédia

Glucose-6-Phosphate dehydrogenase deficiency in children from 0 to 14 years hospitalized at the Pediatric Hospital David Bernardino, Luanda, Angola

The Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common enzymatic defect in the world. The most common clinical manifestations are acute hemolytic anemia associated with drugs, infections, neonatal jaundice and hemolytic non-spherocytic chronic anemia. The main aim of this study was to determine the frequency of major genetic variants of G6PD leading to enzyme deficiency in children from 0 to 14 years at a Pediatric Hospital in Luanda, Angola. A cross-sectional and descriptive analytical study covered a total of 194 children aged from 0 to 14 years, of both genders and hospitalized at the Pediatric Hospital David Bernardino, Luanda between November and December, 2011. The G202A, A376G and C563T mutations of the G6PD gene were determined by real-time PCR with Taqman probes. The disabled A-/A- genotype was detected in 10 girls (10.9%). Among the boys, 21 (20.6%) presented the genotype A-. Considering all the samples, the A- variant was observed in 22.4% of cases. The Mediterranean mutation was not detected in the Angolan sample. Furthermore, no association was found between genotype and anemia, nutritional state and mucosa color. A significant association, however, was observed with jaundice. Based on the results obtained, there is a clear need to identify those with the disabled genotype in the Angolan population in order to avoid cases of drug-induced anemia, particularly in the treatment of malaria, so prevalent in Angola.

Quantificação do valor ambiental em edifícios: estudo de um caso prático

Trabalho Final de Mestrado para obtenção do grau de Mestre em Engenharia Civil

Avaliação por fMRI do córtex visual, motor e auditivo através de estimulação sensoriomotora e sonora em desportistas invisuais e desportistas sem deficiência visual

Mestrado em Radiações Aplicadas às Tecnologias da Saúde - Ramo de especialização: Imagem por Ressonância Magnética

Diagnóstico de avarias em máquinas rotativas utilizando a análise de órbitas

Trabalho Final de Mestrado para obtenção do grau de Mestre em Engenharia Mecânica Perfil Manutenção e Produção