Intermolecular interactions of the malate synthase of Paracoccidioides spp

Background: The fungus Paracoccidioides spp is the agent of paracoccidioidomycosis (PCM), a pulmonary mycosis acquired by the inhalation of fungal propagules. Paracoccidioides malate synthase (PbMLS) is important in the infectious process of Paracoccidioides spp because the transcript is up-regulated during the transition from mycelium to yeast and in yeast cells during phagocytosis by murine macrophages. In addition, PbMLS acts as an adhesin in Paracoccidioides spp. The evidence for the multifunctionality of PbMLS indicates that it could interact with other proteins from the fungus and host. The objective of this study was to identify and analyze proteins that possibly bind to PbMLS (PbMLS-interacting proteins) because protein interactions are intrinsic to cell processes, and it might be possible to infer the function of a protein through the identification of its ligands. Results: The search for interactions was performed using an in vivo assay with a two-hybrid library constructed in S. cerevisiae; the transcripts were sequenced and identified. In addition, an in vitro assay using pull-down GST methodology with different protein extracts (yeast, mycelium, yeast-secreted proteins and macrophage) was performed, and the resulting interactions were identified by mass spectrometry (MS). Some of the protein interactions were confirmed by Far-Western blotting using specific antibodies, and the interaction of PbMLS with macrophages was validated by indirect immunofluorescence and confocal microscopy. In silico analysis using molecular modeling, dynamics and docking identified the amino acids that were involved in the interactions between PbMLS and PbMLS-interacting proteins. Finally, the interactions were visualized graphically using Osprey software. Conclusion: These observations indicate that PbMLS interacts with proteins that are in different functional categories, such as cellular transport, protein biosynthesis, modification and degradation of proteins and signal transduction. These data suggest that PbMLS could play different roles in the fungal cell. © 2013 de Oliveira et al.; licensee BioMed Central Ltd.

Dissection of the role of pili and type 2 and 3 secretion systems in adherence and biofilm formation of an atypical enteropathogenic escherichia coli strain

Atypical enteropathogenic Escherichia coli (aEPEC) strains are diarrheal pathogens that lack bundle-forming pilus production but possess the virulence-associated locus of enterocyte effacement. aEPEC strain 1551-2 produces localized adherence (LA) on HeLa cells; however, its isogenic intimin (eae) mutant produces a diffuse-adherence (DA) pattern. In this study, we aimed to identify the DA-associated adhesin of the 1551-2 eae mutant. Electron microscopy of 1551-2 identified rigid rod-like pili composed of an 18-kDa protein, which was identified as the major pilin subunit of type 1 pilus (T1P) by mass spectrometry analysis. Deletion of fimA in 1551-2 affected biofilm formation but had no effect on adherence properties. Analysis of secreted proteins in supernatants of this strain identified a 150-kDa protein corresponding to SslE, a type 2 secreted protein that was recently reported to be involved in biofilm formation of rabbit and human EPEC strains. However, neither adherence nor biofilm formation was affected in a 1551-2 sslE mutant. We then investigated the role of the EspA filament associated with the type 3 secretion system (T3SS) in DA by generating a double eae espA mutant. This strain was no longer adherent, strongly suggesting that the T3SS translocon is the DA adhesin. In agreement with these results, specific anti-EspA antibodies blocked adherence of the 1551-2 eae mutant. Our data support a role for intimin in LA, for the T3SS translocon in DA, and for T1P in biofilm formation, all of which may act in concert to facilitate host intestinal colonization by aEPEC strains. ©2013, American Society for Microbiology.

Isolamento e caracterização parcial de adesina de Paracoccidioides brasiliensis ligante de fibronectina

Pós-graduação em Biociências e Biotecnologia Aplicadas à Farmácia - FCFAR

Caracterização da enolase de Paracoccidioides brasiliensis e identificação proteômica de novas moléculas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Ocorrência de cepas de Escherichia coli que apresentam o gene de Shiga toxina em queijo mussarela produzido artesanalmente

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Desenvolvimento e caracterização de sistemas nanoestruturados bioadesivos contendo peptídeo análogo à adesina do streptococcus mutans

Pós-graduação em Ciências Farmacêuticas - FCFAR

A independência do solo que habitamos: autonomia, poder e cultura política na construção do império brasileiro. Sergipe (1750-1831)

Pós-graduação em História - FCHS

Clonagem e caracterização da proteína 14-3-3 de Paracoccidioides brasiliensis durante sua interação com células epiteliais

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Análise da expressão gênica global da bactéria Xylella fastidiosa em laranja doce por microarranjos de DNA

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Care to mastectomy with axillary lymphadenectomy, lymphedema prevention: An integrative review

Objective: to examine the care provided to women affected by breast cancer and who has performed axillary lymphadenectomy. Method: integrative review conducted in the databases LILACS and PubMed between 2000 and 2012, leaving the question << What care is provided to prevent lymphedema in women who underwent lymphadenectomy? >> Resulting in 18 articles, analyzed under the following variables: author, title, methodology, year, journal, goal, prevention and control of lymphedema therapies and adherence to rehabilitation. Results: three categories are identified for rehabilitation of patient: prevention and control, therapies and adherence. Conclusion: preoperative is poorly addressed by; emphasizes the importance of home visits as part of rehabilitation, which is effective with the accession of the patient who needs information for its follow-up, no need for a guideline for this type treatment.

Adesão ao valor justiça em crianças e adolescentes

This search is part of a larger study conducted at the Carlos Chagas Foundation (SP-Brazil), which aims to build a scale to assess adhesion to moral values by basic education students and their teachers. This text addresses only the value of justice. The responses of 111 10-13 year old children and 121 teenagers between 14 and 17 years of age at public and private schools in the city of São Paulo (SP-Brazil) were compared. Among the various topics addressed, some issues of distributive, retributive and commutative justice were selected. In the questionnaire, the alternatives offered were built in increasing stages of decentration from a social perspective inspired by Kohlberg. The results confirmed the two hypotheses that had been raised: there was a progression in the choice of the answers concerning the stages between children and teenagers and there were differences between the three forms of justice considered, in relation to the achievement of higher levels.

Biofilme estafilocócico: prevenção, detecção da produção e determinação do perfil de resistência a antimicrobianos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Educação ambiental e contexto escolar: questões para um programa de pesquisas

In this paper are systematized the discussions conducted by GDP "Research in EA and School Context", in VII EPEA, which have had, as its main axis, themes, authors and theoretical perspectives privileged in research presented at the event and declared themselves affiliated to this GDP. The dialogue with researches from previous meetings and other, aims to deepen the questions posed by the research group. The discussions allowed to identify themes and issues that present themselves as significant and challenging for research in EA in the school context, namely the conversion of schools into sustainable educational spaces; public-private partnerships, with great interest from companies and NGOs in school spaces, waving kits with educational materials produced for the school; lack of investment in public policies for teacher training in EA; tendency to an adherence to critical environmental education, without, however, have clear theoretical and methodological commitments under this option.

Efecto del blanqueamiento dental con peroxido de hidrógeno al 35% en la resistencia de unión de dientes restaurados con resina compuesta

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Importance of adhesins in virulence of Paracoccidioides spp.

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)