Browsing by Author "Pedroso, Enio Roberto Pietra"
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Item Combined use of Paracoccidioides brasiliensis recombinant rPb27 and rPb40 antigens in an enzyme-linked immunosorbent assay for immunodiagnosis of paracoccidioidomycosis.(2011) Fernandes, V. C.; Coitinho, Juliana Barbosa; Veloso, Juliana Márcia Ribeiro; Araújo, Stanley de Almeida; Pedroso, Enio Roberto Pietra; Goes, Alfredo Miranda deParacoccidioidomycosis (PCM) is one of themost important endemicmycoses in LatinAmerica; it's usually diagnosed by observation and/or isolation of the etiologic agent, Paracoccidioides brasiliensis, as well as by a variety of immunological methods, such as complement fixation and immunodiffusion. Although these approaches are useful, historically their sensitivity and specificity have often been compromised by the use of complex mixtures of undefined antigens. The use of combinations of purified,well-characterized antigens appears preferable andmay yield good results. In the present study combinations of the previously described 27-kDa recombinant antigen(rPb27) and a recombinant 40-kDa-molecular-mass antigen(rPb40) fromthis fungus, that was identified by Goes et al. (2005) through the AST strategy as a homolog of Neurospora crassa calcineurin B, were used in an indirect enzyme linked immunosorbent assay (ELISA) for diagnosis and follow-up of patients with PCM. The complete coding cDNA of rPb40 and rPb27 were cloned into a pET-21a and a pET-DEST 42 plasmid, respectively, expressed in E. coli with a his-tag and purified by affinity chromatography. Among 109 PCM serum samples analyzed, a homogeneous IgG response to these proteinswas observed. 62 serumsamples frompatientswith other diseases, 18 frompatientswith othermycosis and 23 fromhealthy individualswere also studied. Detection of anti-rPb27 and anti-rPb40 antibodies in sera of patientswith PCMby ELISA using a combination of the two purified proteins showed a sensitivity of 96% with a specificity of 100% in relation to control normal human sera andto sera frompatientswith other systemicmycosis and 93.5%to sera from patients with diverse infections. The use of this two proteins combination provided an excellent immunodiagnosis assaywith great values of sensitivity and specificity, even in relation to sera frompatients with othermycosis,making possible the standadization of a new methodology to diagnose this important mycosis, with a good confiability and reprodutibility.Item Cutaneous disseminated paracoccidioidomycosis.(2012) Araújo, Stanley de Almeida; Espindola, Bernardo Magalhães; Pedroso, Enio Roberto PietraItem Paracoccidioidomycosis case series with and without central nervous system involvement.(2012) Pedroso, Vinicius Sousa Pietra; Lyon, Ana Cláudia; Araújo, Stanley de Almeida; Veloso, Juliana Márcia Ribeiro; Pedroso, Enio Roberto Pietra; Teixeira, Antônio LúcioIntroduction: Paracoccidioidomycosis (PCM) is the most important systemic mycosis in South America. Central nervous system involvement is potentially fatal and can occur in 12.5% of cases. This paper aims to contribute to the literature describing eight cases of neuroparacoccidioidomycosis (NPMC) and compare their characteristics with patients without neurological involvement, to identify unique characteristics of NPCM. Methods: A cohort of 213 PCM cases was evaluated at the Infectious Diseases Clinic of the University Hospital, Federal University of Minas Gerais, Brazil, from October 1976 to August 2008. Epidemiological, clinical, laboratory, therapeutic and follow-up data were registered. Results: Eight patients presented NPCM. The observed NPCM prevalence was 3.8%. One patient presented the subacute form of PCM and the other seven presented the chronic form of the disease. The parenchymatous form of NPCM occurred in all patients. 60% of the patients who proceeded from the north/ northeast region of Minas Gerais State developed NPCM. The neurological involvement of a mother and her son was observed. NPCM patients exhibited demographical and clinical profiles similar to what is described in the literature. When NPCM cases were compared to PCM patients, there were differences in relation to origin and positive PCM family history. Conclusions: The results corroborate the clinical view that the neurological findings are extremely important in the evaluation of PCM patients. Despite the limitations of this study, the differences in relation to patient’s origins and family history point to the need of further studies to determine the susceptibility factors involved in the neurological compromise.