An efficient oxidative stress response is important to the fungal pathogen Paracoccidioides to survive within the human host. In this study, oxidative stress was mimicked by exposure of yeast cells to hydrogen peroxide (2 mM H2O2). To investigate the effect of H2O2 on the proteome of Paracoccidioides, we used a large scale 2-DE protein gel electrophoresis approach to analyze differentially expressed proteins/isoforms that were detected in early (2 h) and in late (6 h) oxidative stress treatments. All proteins/isoforms were grouped based on their functional categories that revealed a global activation of antioxidant enzymes, such as catalase, superoxide dismutase, cytochrome C peroxidase and thioredoxin. A view of the metabolic cell profile, as determined by proteomics, depicted a shift in the yeast cells metabolism as suggested by the activation of the pentose phosphate pathway, a great source of cellular reducing power in the form of NADPH. Additionally, in silico analyzes depicted 34 oxidoreductases proteins/isoforms putatively involved with defense against oxidative stress. Confirmatory assays of enzymatic activity, flow cytometry, transcript levels and NADPH measurements, produced data in agreement with proteomic analysis.
Clinical cases involving paracoccidioidomycosis in children, diagnosed in Mato Grosso State, in the central western region of Brazil, are rare despite the state being classified with a moderate to high incidence. We describe a clinical case of infant acute disseminated Paracoccidioidomycosis in Mato Grosso State, Brazil, highlighting the importance of early differential diagnosis from other severe pathologies, thus contributing to the survival of compromised patients. A 3 year-old male child, weighing 12.8 Kg, originating from Vila Rica, MT, Brazil. The patient presented intermittent 40 masculineC fever evolving over 40 days, dry cough and painless bilateral cervical adenomegaly, showing no signs of inflammation. This was associated with diarrhea, distension and important abdominal pain and weight loss. Diagnosis was achieved by visualization of Paracoccidioides brasiliensis yeasts in a direct mycological exam and posterior fungus isolation in culture medium. The patient evolved presenting good clinical response to antifungal treatment and progressive reduction of abdominal and cervical ganglions. To improve the prognosis of compromised patients it is essential that professionals realize a full clinical-laboratorial evaluation, including differential diagnoses for other severe pathologies, as early as possible. The degree and intensity of paracoccidioidomycosis compromise are determining factors for defining the most efficient treatment.
Este trabalho teve como objetivo determinar o perfil imunoquímico de três exoantígenos, obtidos de amostras clínicas de Paracoccidioides lutzii procedentes do estado de Mato Grosso. Para a obtenção dos exoantígenos, seguiu-se o protocolo padronizado por Camargo et al. e o perfil imunoquímico foi obtido com o emprego da técnica de SDS Page. As frações observadas demonstraram pesos moleculares de 20, 30 e 130 kDa; a glicoproteina de 130 kDa foi evidenciada tendo por base os três exoantígenos utilizados. A detecção desta fração de alto peso molecular sugere um perfil imunoquímico distinto, o que pode contribuir para elucidar as reações falso-negativas obtidas com o antígeno usualmente utilizado (obtido da cepa de referência B-339 de P. brasiliensis). Este achado assume grande importância no sentido de aumentar a especificidade das reações sorológicas utilizadas, para monitorar o prognóstico dos pacientes acometidos pela paracoccidioidomicose.
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