We identified and characterized two chitinases, named MpCHIT1 and MpCHIT2, from the fungus Moniliophthora perniciosa - the etiologic agent of witches' broom disease in cacao tree (Theobroma cacao L.) - during its development, mainly in the mycelia phases preceding the basidioma formation. The expression of MpCHIT1 and MpCHIT2, together with MpCHS and MpATG8 (chitin synthase and autophagy genes, respectively), was analyzed during the M. perniciosa growth and development on bran-based solid medium as well as in liquid medium containing H2O2 or rapamycin (oxidative and nutritional related-autophagy stress agents, respectively). In order to link the expression of chitin metabolism-related genes to nutritional composition influencing fungus development, we also quantified total and reduced sugars, as well as macro- and micronutrients in the bran-based solid medium. The expression analysis showed that the MpCHS expression increased through mycelial development and then decreased in the primordium and basidioma phases, while the expression of MpCHIT1 and MpCHIT2 was higher in basidioma and primordium phases, respectively. Moreover, the expression pattern of MpCHIT1 and MpCHIT2 is distinct, the second correlated with the MpATG8 expression pattern and possibly with autophagy process, while the first may be related to the basidioma formation. The quantification of total and reduced sugars, as well as macro- and micronutrients supported the idea that the cell wall restructuration due to MpCHS, MpCHIT1 and MpCHIT2 is related to stress and fungal nutrient reallocation, allowing the formation and development of the basidioma. Experiments involving M. perniciosa growth on liquid medium containing H2O2 or rapamycin showed that MpCHIT1 and MpCHIT2 were over-expressed in response to oxidative but also to nutritional related-autophagy stresses. Interestingly, the expression level of MpCHS, MpCHIT1 and MpCHIT2 in presence of rapamycin is similar to the one observed in the primordium and basidioma from bran-based solid medium. The analysis of the overall data allowed designing a general scheme of chitin metabolism and autophagy during M. perniciosa development, focusing on the mycelium phases as crucial and environmentally influenced steps preceding the primordium and basidioma formation. These data support the idea that the nutritional environment of M. perniciosa influences its development and life cycle.
A terapia fotodinâmica (TFD) é um tratamento antifúngico que se destaca por não induzir resistência. O estudo objetivou avaliar os efeitos da TFD em modelo experimental de artrite articular induzida por Paracoccidiodes brasiliensis. Para isto Ratos Wistar foram distribuídos em grupos e inoculados com P. brasiliensis ou PBS (controle) na articulação do joelho. Animais dos grupos TFD40 e TFD120 receberam tratamento com TFD 40 J/cm 2 e 120 J/cm 2 , respectivamente; o grupo MB, tratado com azul de metileno; e os grupos laser, L40 e L120, tratados com 40 J/cm 2 e 120 J/cm 2 , respectivamente. Foi observado que a formação de edema, titulação de anticorpos anti-Gp43 e o índice de artrite foram significativas nos grupos inoculados e menos evidentes nos grupos TFD40 e TFD120. Avaliação anatomopatológica do grupo TFD120 evidenciou melhora nos aspectos morfológicos da membrana sinovial, em comparação aos demais grupos inoculados. Conclui-se que TFD de 120 J/cm 2 reduz a artrite de forma mais efetiva, o que pode ter um potencial impacto no auxílio ao tratamento de P. brasiliensis.
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