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

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

Soroprevalência da infecção pelo vírus da hepatite B em indivíduos com evidência clínica de hepatite em Goiânia, Goiás: detecção do DNA viral e determinação dos subtipos

The presence of serological markers for hepatitis B virus (HBsAg, anti-HBc IgM and Anti-HBc total) was investigated in the serum of 1,396 individuals who had clinical suspect of hepatitis. It was observed that 50.7% of the individuals were positive and, from the total of the studied individuals, 14.5% were positive for HBsAg. From these, 8.5% were also positive for anti-HBc IgM. The analysis in relation to gender showed a higher seroprevalence index among male individuals (p < 0.0001). It was observed the occurrence of subtypes adw2 (62.7%), ayw3 (23.5%), ayw2 (9.8%) and adw4 (3.9%). The viral DNA was detected in 61 (33.9%) HBsAg positive samples and in one sample positive only for anti-HBc total. These results indicate an important incidence of the HBV infection in this population, and reinforce previous studies regarding this virus in the central west region of Brazil.Investigou-se a presença dos marcadores sorológicos AgHBs, anti-HBc IgM e anti-HBc total no soro de 1.396 indivíduos com suspeita clínica de hepatite. Observou-se uma soroprevalência para a infecção pelo VHB de 50,7%, sendo que do total dos 1.396 indivíduos, 14,5% eram positivos para AgHBs e 8,5% eram também positivos para anti-HBc IgM, tendo ainda sido observado que a soroprevalência foi maior em indivíduos do sexo masculino (p < 0,0001). Por subtipagem foram identificados os seguintes subtipos: adw2 (62,7%), ayw3 (23,5%), ayw2 (9,8%) e adw4 (3,9%). O DNA viral foi detectado em 61 (33,9%) amostras positivas para o AgHBs e em uma amostra positiva somente para anti-HBc total. Estes resultados indicam importante índice de ocorrência da infecção pelo VHB nesta população e reforça dados de estudos anteriores a respeito da importante circulação do vírus na região Centro-Oeste do Brasil

The malate synthase of Paracoccidioides brasiliensis is a linked surface protein that behaves as an anchorless adhesin

BACKGROUND: The pathogenic fungus Paracoccidioides brasiliensis is the agent of paracoccidioidomycosis (PCM). This is a pulmonary mycosis acquired by inhalation of fungal airborne propagules that can disseminate to several organs and tissues leading to a severe form of the disease. Adhesion and invasion to host cells are essential steps involved in the internalization and dissemination of pathogens. Inside the host, P. brasiliensis may use the glyoxylate cycle for intracellular survival. RESULTS: Here, we provide evidence that the malate synthase of P. brasiliensis (PbMLS) is located on the fungal cell surface, and is secreted. PbMLS was overexpressed in Escherichia coli, and polyclonal antibody was obtained against this protein. By using Confocal Laser Scanning Microscopy, PbMLS was detected in the cytoplasm and in the cell wall of the mother, but mainly of budding cells of the P. brasiliensis yeast phase. PbMLSr and its respective polyclonal antibody produced against this protein inhibited the interaction of P. brasiliensis with in vitro cultured epithelial cells A549. CONCLUSION: These observations indicated that cell wall-associated PbMLS could be mediating the binding of fungal cells to the host, thus contributing to the adhesion of fungus to host tissues and to the dissemination of infection, behaving as an anchorless adhesin