Jogos digitais educacionais como uma proposta de trabalho em educação ambiental

Pereira, R.; Gouveia, L. e Dinis, A. (2021). Jogos digitais educacionais como uma proposta de trabalho em Educação Ambiental. Seminário Doutoramento em Ciência da Informação, especialidade Sistemas, Tecnologias e Gestão da Informação (SITEGI). 16 de Julho. Universidade Fernando Pessoa. Porto.info:eu-repo/semantics/publishedVersio

A secreted serine protease of Paracoccidioides brasiliensis and its interactions with fungal proteins

<p>Abstract</p> <p>Background</p> <p><it>Paracoccidioides brasiliensis </it>is a thermodimorphic fungus, the causative agent of paracoccidioidomycosis (PCM). Serine proteases are widely distributed and this class of peptidase has been related to pathogenesis and nitrogen starvation in pathogenic fungi.</p> <p>Results</p> <p>A cDNA (<it>Pb</it>sp) encoding a secreted serine protease (<it>Pb</it>SP), was isolated from a cDNA library constructed with RNAs of fungal yeast cells recovered from liver of infected mice. Recombinant <it>Pb</it>SP was produced in <it>Escherichia coli</it>, and used to develop polyclonal antibodies that were able to detect a 66 kDa protein in the <it>P. brasiliensis </it>proteome. <it>In vitro </it>deglycosylation assays with endoglycosidase H demonstrated that <it>Pb</it>SP is a <it>N</it>-glycosylated molecule. The <it>Pb</it>sp transcript and the protein were induced during nitrogen starvation. The <it>Pb</it>sp transcript was also induced in yeast cells infecting murine macrophages. Interactions of <it>Pb</it>SP with <it>P. brasiliensis </it>proteins were evaluated by two-hybrid assay in the yeast <it>Saccharomyces cerevisiae</it>. <it>Pb</it>SP interacts with a peptidyl prolyl cis-trans isomerase, calnexin, HSP70 and a cell wall protein PWP2.</p> <p>Conclusions</p> <p>A secreted subtilisin induced during nitrogen starvation was characterized indicating the possible role of this protein in the nitrogen acquisition. <it>Pb</it>SP interactions with other <it>P. brasiliensis </it>proteins were reported. Proteins interacting with <it>Pb</it>SP are related to folding process, protein trafficking and cytoskeleton reorganization.</p

The transcriptome analysis of early morphogenesis in Paracoccidioides brasiliensis mycelium reveals novel and induced genes potentially associated to the dimorphic process

BACKGROUND: Paracoccidioides brasiliensis is a human pathogen with a broad distribution in Latin America. The fungus is thermally dimorphic with two distinct forms corresponding to completely different lifestyles. Upon elevation of the temperature to that of the mammalian body, the fungus adopts a yeast-like form that is exclusively associated with its pathogenic lifestyle. We describe expressed sequence tags (ESTs) analysis to assess the expression profile of the mycelium to yeast transition. To identify P. brasiliensis differentially expressed sequences during conversion we performed a large-scale comparative analysis between P. brasiliensis ESTs identified in the transition transcriptome and databases. RESULTS: Our analysis was based on 1107 ESTs from a transition cDNA library of P. brasiliensis. A total of 639 consensus sequences were assembled. Genes of primary metabolism, energy, protein synthesis and fate, cellular transport, biogenesis of cellular components were represented in the transition cDNA library. A considerable number of genes (7.51%) had not been previously reported for P. brasiliensis in public databases. Gene expression analysis using in silico EST subtraction revealed that numerous genes were more expressed during the transition phase when compared to the mycelial ESTs [1]. Classes of differentially expressed sequences were selected for further analysis including: genes related to the synthesis/remodeling of the cell wall/membrane. Thirty four genes from this family were induced. Ten genes related to signal transduction were increased. Twelve genes encoding putative virulence factors manifested increased expression. The in silico approach was validated by northern blot and semi-quantitative RT-PCR. CONCLUSION: The developmental program of P. brasiliensis is characterized by significant differential positive modulation of the cell wall/membrane related transcripts, and signal transduction proteins, suggesting the related processes important contributors to dimorphism. Also, putative virulence factors are more expressed in the transition process suggesting adaptation to the host of the yeast incoming parasitic phase. Those genes provide ideal candidates for further studies directed at understanding fungal morphogenesis and its regulation

Identification and characterization of Tc1/mariner-like DNA transposons in genomes of the pathogenic fungi of the Paracoccidioides species complex

<p>Abstract</p> <p>Background</p> <p><it>Paracoccidioides brasiliensis </it>(Eukaryota, Fungi, Ascomycota) is a thermodimorphic fungus, the etiological agent of paracoccidioidomycosis, the most important systemic mycoses in Latin America. Three isolates corresponding to distinct phylogenetic lineages of the <it>Paracoccidioides </it>species complex had their genomes sequenced. In this study the identification and characterization of class II transposable elements in the genomes of these fungi was carried out.</p> <p>Results</p> <p>A genomic survey for DNA transposons in the sequence assemblies of <it>Paracoccidioides</it>, a genus recently proposed to encompass species <it>P. brasiliensis </it>(harboring phylogenetic lineages S1, PS2, PS3) and <it>P. lutzii </it>(<it>Pb01-like </it>isolates), has been completed. Eight new <it>Tc1/mariner </it>families, referred to as Trem (<b>Tr</b>ansposable <b>e</b>lement <b>m</b>ariner), labeled A through H were identified. Elements from each family have 65-80% sequence similarity with other <it>Tc1/mariner </it>elements. They are flanked by 2-bp TA target site duplications and different termini. Encoded DDD-transposases, some of which have complete ORFs, indicated that they could be functionally active. The distribution of Trem elements varied between the genomic sequences characterized as belonging to <it>P. brasiliensis </it>(S1 and PS2) and <it>P. lutzii</it>. TremC and H elements would have been present in a hypothetical ancestor common to <it>P. brasiliensis </it>and <it>P. lutzii</it>, while TremA, B and F elements were either acquired by <it>P. brasiliensis </it>or lost by <it>P. lutzii </it>after speciation. Although TremD and TremE share about 70% similarity, they are specific to <it>P. brasiliensis </it>and <it>P. lutzii</it>, respectively. This suggests that these elements could either have been present in a hypothetical common ancestor and have evolved divergently after the split between <it>P. brasiliensis </it>and <it>P. Lutzii</it>, or have been independently acquired by horizontal transfer.</p> <p>Conclusions</p> <p>New families of <it>Tc1/mariner </it>DNA transposons in the genomic assemblies of the <it>Paracoccidioides </it>species complex are described. Families were distinguished based on significant BLAST identities between transposases and/or TIRs. The expansion of Trem in a putative ancestor common to the species <it>P. brasiliensis </it>and <it>P. lutzii </it>would have given origin to TremC and TremH, while other elements could have been acquired or lost after speciation had occurred. The results may contribute to our understanding of the organization and architecture of genomes in the genus <it>Paracoccidioides</it>.</p