Identification of Critical Amino Acids in an Immunodominant IgE Epitope of Pen c 13, a Major Allergen from Penicillium citrinum

Background: Pen c 13, identified as a 33-kDa alkaline serine protease, is a major allergen secreted by Penicillium citrinum. Detailed knowledge about the epitopes responsible for IgE binding would help inform the diagnosis/prognosis of fungal allergy and facilitate the rational design of hypoallergenic candidate vaccines. The goal of the present study was to characterize the IgE epitopes of Pen c 13. Methodology/Principal Findings: Serum samples were collected from 10 patients with mold allergy and positive Pen c 13 skin test results. IgE-binding epitopes on rPen c 13 were mapped using an enzymatic digestion and chemical cleavage method, followed by dot-blotting and mass spectrometry. A B-cell epitope-predicting server and molecular modeling were used to predict the residues most likely involved in IgE binding. Theoretically predicted IgE-binding regions were further confirmed by site-directed mutagenesis assays. At least twelve different IgE-binding epitopes located throughout Pen c 13 were identified. Of these, peptides S16 (A 148 –E 166) and S22 (A 243 –K 274) were recognized by sera from 90 % and 100 % of the patients tested, and were further confirmed by inhibition assays. Peptide S22 was selected for further analysis of IgE-binding ability. The results of serum screening showed that the majority of IgE-binding ability resided in the C-terminus. One Pen c 13 mutant, G270A (T 261 –K 274), exhibited clearly enhanced IgE reactivity, whereas another, K274A, exhibited dramatically reduced IgE reactivity

Analisando as pesquisas em educação especial no Brasil Analysing research in special education in Brazil

Nosso objetivo foi examinar a articulação lógica entre o problema e a proposição teórico-metodológica das produções na área da Educação Especial, focando os seus pressupostos epistemológicos. Nos fundamentamos nos pressupostos das tendências empírico-analítica, fenomenológica-hermenêutica, crítico-dialética e do paradigma da complexidade. O procedimento adotado foi interpretar todas as dissertações/teses produzidas nos Programas de Pós-Graduação em Educação e Educação Especial do Brasil, que versam sobre Educação Especial, produzidas nos anos de 2001, 2002 e 2003, disponíveis no banco de teses da CAPES. Encontramos as tendências empírica, fenomenológica e dialética. Os equívocos encontrados foram a não inserção da pesquisa entre as produções na área; ausência de criticidade; não posicionamento numa determinada concepção de educação; construção teórica fundamentada em concepções diferentes; falta de coerência nos pressupostos teórico-metodológicos; não explicitação metodológica; não descrição dos procedimentos éticos; e má elaboração dos resumos. Concluímos pela necessidade da melhoria das dissertações/teses para que possamos avançar na produção de conhecimento na área da Educação Especial.<br>Our objective was to analyze the logical articulation between the problem and the theoretical-methodological proposal of studies in the field of Special Education, focusing on the epistemological issues. We based our study on the empiric-analytical tendencies, phenomenology-hermeneutic, critical-dialectical and the complexity paradigm. The procedure that was adopted was interpreting all dissertations/thesis produced in Post-Graduate programs in Education and Special Education in Brazil, which focus on Special Education, produced in 2001, 2002 and 2003, available online at CAPES' thesis database. We found empirical, phenomenological and dialectic tendencies. The errors encountered included the failure to include the research among the productions in the field; lack of critical approach; lack of making explicit what educational conception the study was based on; theoretical construction based on different conceptions; lack of coherence in the theoretical-methodological proposals; lack of methodological specification; absence of ethical procedural descriptions; and poorly written abstracts. We came to the conclusion that improvements in theses /dissertations are necessary so as to continually move forward in the production of knowledge in the field of Special Education