Development of a standard operation procedure (SOP) for the polyphasic identification of clinically relevant species

The identification of dermatophytes and other clinically relevant filamentous fungi has been essential for a rapid diagnosis and the preservation of authentic microorganisms in culture collections. The aim of this work was the development of a Standard Operating Procedure (SOP) for the polyphasic identification of clinically important species that might be deposited in Micoteca da Universidade do Minho (MUM) and can be used in different clinical settings. To achieve this goal 27 strains of dermatophytes belongs to the taxa Epidermophyton floccosum, Microsporum spp. and Trichophyton spp. and 11 strains of Sporothrix genus belonging to different species were characterised using microscopy, biochemical tests and molecular biology. The microscopic analysis allowed distinguishing the genera of dermatophytes, since different genera have different macroaleuriospores and microaleuriospores. On macroscopic examination, it is verifiable that, in PDA medium, T. rubrum had a red-brown pigment (Sudman et al., 1965). On the other hand, T. violaceum suffered a growth inhibition in TA-1 medium (David Ellis et al., 2007). As for Sporothrix spp., it was not possible to distinguish the species through micro and macroscopic analyses. Through the Urease Production Test, it was possible to verify that T. tonsurans and T. mentagrophytes produced a high amount of the enzyme and T. violaceum and T. rubrum strains did not. In the case of Sporothrix spp., it was possible to verify that S. mexicana, S. pallida, S. schenckii and S. brasiliensis produced a high amount of urease and catalase. The strains S. inflata, S. fungorum, S. ghanensis and S. globosa neither produced urease or catalase. The present project allowed developing an effective and fast procedure in the identification of dermatophytes, however, phenotypic analysis is quite limiting in identifying strains (mainly from the Sporothrix spp. complex) and, therefore, the genotypic approach is essential for unambiguous identification.info:eu-repo/semantics/publishedVersio

A Constituição Federal de 1988 nas charges do Correio Braziliense

O artigo analisa as charges publicadas no jornal Correio Braziliense sobre a promulgação da Constituição Federal de 1988. Utilizando como metodologia a análise crítica da narrativa, tal como proposta por Motta (2013), o objetivo é investigar os significados e efeitos de sentido produzidos e compreender como essas narrativas interpretaram esse acontecimento histórico. Como resultado, o trabalho mostra que as charges fornecem elementos importantes para compreendermos os bastidores, as representações, posições e ideias durante o processo histórico de redemocratização e elaboração do novo texto constitucional brasileiro. Por meio da análise crítica da narrativa percebe-se que, embora a primeira charge do Correio Braziliense represente a Constituição como um símbolo maior de esperança pela busca de um Estado democrático de direito e uma nova chance de transformação, com o desenrolar dos fatos e os novos episódios, a Constituição de 1988 nas páginas do Correio Braziliense também começou a ganhar traços que permitissem o questionamento de sua obediência, bem como sua funcionalidade e futuro. Incertezas se o Estado seria realmente capaz de cumprir com as promessas constitucionais. O artigo também aponta que as charges vão muito além do humor ao evidenciarem as manobras políticas da época. Elas têm o papel de informar e vão se apoiar sempre em uma narrativa maior, que são as notícias diárias

Jornalismo, redes sociais e cidadania: Mediação e recepção no caso “ladrão e vacilão”

O artigo analisa em que aspectos a cultura brasileira influencia o debate sobre questões sociais e políticas no Brasil e sua influência na cidadania. Utilizando como objeto de estudo o caso do adolescente que teve a frase "eu sou ladrão e vacilão" tatuada em sua testa, a análise hermenêutica nos mostrou que a origem profunda da violência no Brasil e a instabilidade política refletiram nas mensagens produzidas pelo públic

In silico characterization of microRNAs-like sequences in the genome of Paracoccidioides brasiliensis

Eukaryotic cells have different mechanisms of post-transcriptional regulation. Among these mechanisms, microRNAs promote regulation of targets by cleavage or degradation of the mRNA. Fungi of the Paracoccidioides complex are the etiological agents of the main systemic mycosis of Latin America. These fungi present a plasticity to adapt and survive in different conditions, and the presence of microRNAs-like molecules could be part of the mechanisms that provide such plasticity. MicroRNAs produced by the host influence the progression of this mycosis in the lungs besides regulating targets involved in apoptosis in macrophage, activation of T and B cells and the production of cytokines. Therefore, this work analyzed the presence of regions in the genome of this fungus with a potential to encode microRNAs-like molecules. Here we show by analysis of sequence similarity the presence of 18 regions, putatively coding for microRNAs-like molecules in the Paracoccidioides brasiliensis genome. We also described the conservation of dicer and argonaut proteins and the cognate transcripts induced in the yeast parasitic phase. This work represents a starting point for the analysis of the presence of those molecules in the morphological stages of the fungus and their role in fungal development

Indicadores de resultado clinicamente úteis no contexto das estratégias terapêuticas promotoras da adaptação em doentes com cancro da mama

Neste artigo descreve-se o processo inicial de identificação de um painel de indicadores de resultado, clinicamente úteis, capazes de promover a monitorização dos processos de adaptação, em doentes com cancro da mama. A doença oncológica da mama constitui um agente de stress capaz de ameaçar a integridade psicológica e física da pessoa, representando um impacto violento no bem-estar e na qualidade de vida. Como metodologia utilizou-se uma abordagem qualitativa e exploratória, com recurso a revisão bibliográfica e a entrevistas exploratórias a doentes com cancro da mama, que pertencem a um grupo de auto-ajuda, e à avaliação por peritos. Numa primeira abordagem, de acordo com a revisão da literatura efectuada, podemos inferir que um estilo de coping mais activo sob a forma de espírito de luta, suporte social, autocontrolo, índices favoráveis de optimismo e dimensão espiritual presente, induz padrões de respostas mais adaptativas. A partir dos resultados das entrevistas exploratórias e da revisão da literatura, evoluímos na identificação de um conjunto bastante alargado de indicadores de resultado de enfermagem, utilizando a Classificação de Resultados de Enfermagem. O referido conjunto de indicadores foi então submetido a um painel de peritos, neste domínio problemático. De acordo com as opções dos peritos, fomos capazes de chegar a um painel de 110 indicadores de resultado, potencialmente úteis para a monitorização e acompanhamento das doentes que experimentam uma situação de doença oncológica da mama. Concluímos que é essencial para a disciplina de enfermagem a identificação de indicadores de resultado, úteis e mensuráveis, que permitam avaliar a assistência em saúde, como meio de sustentar a qualidade do exercício profissional dos enfermeiros.info:eu-repo/semantics/publishedVersio

Comparative Proteomic Analysis of Histoplasma capsulatum Yeast and Mycelium Reveals Differential Metabolic Shifts and Cell Wall Remodeling Processes in the Different Morphotypes

Histoplasma capsulatum is a thermally dimorphic fungus distributed worldwide, but with the highest incidence in the Americas within specific geographic areas, such as the Mississippi River Valley and regions in Latin America. This fungus is the etiologic agent of histoplasmosis, an important life-threatening systemic mycosis. Dimorphism is an important feature for fungal survival in different environments and is related to the virulence of H. capsulatum, and essential to the establishment of infection. Proteomic profiles have made important contributions to the knowledge of metabolism and pathogenicity in several biological models. However, H. capsulatum proteome studies have been underexplored. In the present study, we report the first proteomic comparison between the mycelium and the yeast cells of H. capsulatum. Liquid chromatography coupled to mass spectrometry was used to evaluate the proteomic profile of the two phases of H. capsulatum growth, mycelium, and yeast. In summary, 214 and 225 proteins were only detected/or preferentially abundant in mycelium or yeast cells, respectively. In mycelium, enzymes related to the glycolytic pathway and to the alcoholic fermentation occurred in greater abundance, suggesting a higher use of anaerobic pathways for energy production. In yeast cells, proteins related to the tricarboxylic acid cycle and response to temperature stress were in high abundance. Proteins related to oxidative stress response or involved with cell wall metabolism were identified with differential abundance in both conditions. Proteomic data validation was performed by enzymatic activity determination, Western blot assays, or immunofluorescence microscopy. These experiments corroborated, directly or indirectly, the abundance of isocitrate lyase, 2-methylcitrate synthase, catalase B, and mannosyl-oligosaccharide-1,2-alpha-mannosidase in the mycelium and heat shock protein (HSP) 30, HSP60, glucosamine-fructose-6-phosphate aminotransferase, glucosamine-6-phosphate deaminase, and N-acetylglucosamine-phosphate mutase in yeast cells. The proteomic profile-associated functional classification analyses of proteins provided new and interesting information regarding the differences in metabolism between the two distinct growth forms of H. capsulatum

Cell Wall Synthesis, Development of Hyphae and Metabolic Pathways Are Processes Potentially Regulated by MicroRNAs Produced Between the Morphological Stages of Paracoccidioides brasiliensis

MicroRNAs are molecules involved in post-transcriptional gene regulation. In pathogenic fungi, microRNAs have been described at different morphological stages by regulating targets involved in processes such as morphogenesis and energy production. Members of the Paracoccidioides complex are the main etiological agents of a systemic mycosis in Latin America. Fungi of the Paracoccidioides complex present a wide range of plasticity to colonize different niches. In response to environmental changes these fungi undergo a morphological switch, remodel their cellular metabolism and modulate structural cell wall components. However, the underlying mechanisms regulating the gene expression is not well understood. By using high performance sequencing and bioinformatics analyses, this work characterizes microRNAs produced by Paracoccidioides brasiliensis. Here, we demonstrated that the transcript encoding proteins involved in microRNA biogenesis were differentially expressed in each morphological stage. In addition, 49 microRNAs were identified in cDNA libraries with 44 differentially regulated among the libraries. Sixteen microRNAs were differentially regulated in comparison to the mycelium in the mycelium-to-yeast transition phase. The yeast parasitic phase revealed a complete remodeling of the expression of these small RNAs. Analyses of targets of the induced microRNAs, from the different libraries, revealed that these molecules may potentially regulate in the cell wall, by repressing genes involved in the synthesis and degradation of glucans and chitin. Furthermore, mRNAs involved in cellular metabolism and development were predicted to be regulated by microRNAs. Therefore, this work describes a putative post transcriptional regulation, mediated by microRNAs in P. brasiliensis and its influence on the adaptive processes of thermal dimorphic fungus