Evaluation of the Allergenicity Potential of TcPR-10 Protein from Theobroma cacao

Background: The pathogenesis related protein PR10 (TcPR-10), obtained from the Theobroma cacao-Moniliophthora perniciosa interaction library, presents antifungal activity against M. perniciosa and acts in vitro as a ribonuclease. However, despite its biotechnological potential, the TcPR-10 has the P-loop motif similar to those of some allergenic proteins such as Bet v 1 (Betula verrucosa) and Pru av 1 (Prunus avium). The insertion of mutations in this motif can produce proteins with reduced allergenic power. The objective of the present work was to evaluate the allergenic potential of the wild type and mutant recombinant TcPR-10 using bioinformatics tools and immunological assays. Methodology/Principal Findings: Mutant substitutions (T10P, I30V, H45S) were inserted in the TcPR-10 gene by sitedirected mutagenesis, cloned into pET28a and expressed in Escherichia coli BL21(DE3) cells. Changes in molecular surface caused by the mutant substitutions was evaluated by comparative protein modeling using the three-dimensional structure of the major cherry allergen, Pru av 1 as a template. The immunological assays were carried out in 8-12 week old female BALB/c mice. The mice were sensitized with the proteins (wild type and mutants) via subcutaneous and challenged intranasal for induction of allergic airway inflammation. Conclusions/Significance: We showed that the wild TcPR-10 protein has allergenic potential, whereas the insertion of mutations produced proteins with reduced capacity of IgE production and cellular infiltration in the lungs. On the other hand, in vitro assays show that the TcPR-10 mutants still present antifungal and ribonuclease activity against M. perniciosa RNA. In conclusion, the mutant proteins present less allergenic potential than the wild TcPR-10, without the loss of interesting biotechnological properties. (Résumé d'auteur

A New Approach for Heparin Standardization: Combination of Scanning UV Spectroscopy, Nuclear Magnetic Resonance and Principal Component Analysis

The year 2007 was marked by widespread adverse clinical responses to heparin use, leading to a global recall of potentially affected heparin batches in 2008. Several analytical methods have since been developed to detect impurities in heparin preparations; however, many are costly and dependent on instrumentation with only limited accessibility. A method based on a simple UV-scanning assay, combined with principal component analysis (PCA), was developed to detect impurities, such as glycosaminoglycans, other complex polysaccharides and aromatic compounds, in heparin preparations. Results were confirmed by NMR spectroscopy. This approach provides an additional, sensitive tool to determine heparin purity and safety, even when NMR spectroscopy failed, requiring only standard laboratory equipment and computing facilities

Conhecimento de mulheres de uma Unidade Básica de Saúde da cidade de Natal/RN sobre o exame de Papanicolau Conocimiento de mujeres de una Unidad Básica de Salud de la ciudad de Natal/RN sobre el examen de Papanicolau Knowledge of women about the Paptest in a Basica Unit of Health in Natal

Trata-se de uma pesquisa descritiva quantitativa, desenvolvida em uma Unidade Básica de Saúde em Natal/RN, objetivando identificar o conhecimento de mulheres quanto à importância, à freqüência do exame de Papanicolau, bem como seus cuidados antes de realizá-lo e causas que levam mulheres a não se submeterem a tal exame. Utilizou-se uma entrevista estruturada na coleta de dados antes da consulta ginecológica, com uma amostra intencional de 120 mulheres. Os resultados mostram que as pesquisadas conhecem a importância do exame, a maioria realiza-o anualmente e, no geral, apresentam conhecimento satisfatório sobre os cuidados antes do exame. A vergonha de fazer o exame de Papanicolau e o medo do seu resultado são as principais causas atribuídas para a sua não realização. Conclui-se que os projetos educativos sejam direcionados para a importância, a freqüência e os cuidados necessários antes do exame de Papanicolau, como também, para a interação profissional-cliente durante a consulta ginecológica, visando a reduzir a vergonha e o medo dessas mulheres.<br>Se trata de una investigación descriptiva cuantitativa, desarrollada en una Unidad Básica de Salud en Natal/RN, con el objetivo de identificar el conocimiento de las mujeres en cuanto a la importancia y frecuencia del examen de Papanicolau, así como sus cuidados antes de realizárselo y causas que las llevan a no someterse a tal examen. Se utilizó una entrevista estructurada en la recolección de datos antes de la consulta ginecológica, con una muestra intencional de 120 mujeres. Los resultados muestran que las investigadas conocen la importancia del examen, la mayoría lo realiza anualmente y, en general, presentan conocimiento satisfactorio sobre los cuidados antes del examen. La vergüenza de hacerse el examen de Papanicolau y el miedo de su resultado son las principales causas atribuídas para su no realización. Se concluye que los proyectos educativos estén orientados a la importancia, frecuencia y cuidados necesarios antes del examen de Papanicolau, así como también, a la interacción profesional-cliente durante la consulta ginecológica, visando reducir la vergüenza y el miedo de esas mujeres.<br>This is a descriptive quantitative research carried out at a Basic Health Unit in Natal, State of Rio Grande do Norte, aimed at measuring the knowledge that women have on the importance of the Pap test and the frequency in which they undertake it, as well as how they prepare themselves to undertake it and the causes that lead women to not submit to it. For data collection a structured interview prior to the gynecologic consultation was used, with an intentional sample of 120 women. The results show that the surveyed women are aware of the importance of the test, that most of them undertake it annually and that they generally have satisfactory knowledge as for the care they must take prior to undertaking it. Embarrassment and fear of the results are the main causes associated with the refusal to undertake the test. It can be concluded that the educative projects are to be focused on the importance, the frequency and the care that must be taken prior to the test, as well as on the professional-client interaction during the gynecologic consultation, with the aim of reducing women's embarrassment and fear