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.

Validation of a reference control for an SYBR-Green fluorescence assay-based real-time PCR for detection of bovine herpesvirus 5 in experimentally exposed bovine embryos

The objective of this study was to optimize an internal control to improve SYBR-Green-based qPCR to amplify/detect the BoHV-5 US9 gene in bovine embryos produced invitro and experimentally exposed to the virus. We designed an SYBR-Green-based binding assay that is quick to perform, reliable, easily optimized and compares well with the published assay. Herein we demonstrated its general applicability to detect BoHV-5 US9 gene in bovine embryos produced invitro experimentally exposed to BoHV-5. In order to validate the assay, three different reference genes were tested; and the histone 2a gene was shown to be the most adequate for normalizing the qPCR reaction, by considering melting and standard curves ( p<0.05). On the other hand, no differences were found in the development of bovine embryos invitro whether they were exposed to BoHV-5 reference and field strains comparing to unexposed embryos. The developed qPCR assay may have important field applications as it provides an accurate BoHV-5 US9 gene detection using a proven reference gene and is considerably less expensive than the TaqMan qPCR currently employed in sanitary programs. © 2013 Elsevier Ltd.

Proposta de metodologia para o gerenciamento de fonte radioativa e estimativa de dose efeiva em pacientes submetidos à terapia com iodeto de sódio I-131

Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB

Influência do ganho de massa corporal, induzido por dieta rica em sacarose, em parâmetros biométricos, bioquímicos e biofísicos de ratos Wistar

Pós-graduação em Ciências Fisiológicas - FOA

Análise das propriedades reológicas de materiais cimentícios associando o conceito de empacotamento de partículas

Pós-graduação em Engenharia Mecânica - FEIS

Aplicação da transformada integral generalizada no escoamento potencial em contrações

Pós-graduação em Engenharia Mecânica - FEIS

Estudo do crescimento ósseo de bovinos da raça nelore até 24 meses de idade: padronização e correlação da densidade mineral óssea do (DMD) do III metacarpo com sexo, idade e peso

Pós-graduação em Medicina Veterinária - FMVZ

Avaliação técnica para reativação de uma PCH na Fazenda Experimental Lageado, Botucatu - SP

Pós-graduação em Agronomia (Energia na Agricultura) - FCA

Programa computacional para análise do faturamento do consumo de energia elétrica através do k-hiperbolóide de carga e potência em empresas frigoríficas abatedoras de bovinos

Pós-graduação em Agronomia (Energia na Agricultura) - FCA

A tecnologia informática como auxílio no ensino de geometria para deficientes visuais

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Tratamento da endometriose peritoneal com injeção local de ácido acetilsalicítico: estudo experimental em coelhas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Densidade mineral óssea de vértebras de ratos wistar suspensos pela cauda por 15 e 36 dias

Pós-graduação em Ciência Animal - FMVA

Efeito do ultra-som de baixa potência na reparação óssea em ratos sob ausência de carga: análise densitométrica e biomecânica

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

Influência do laser terapêutico no reparo de defeito ósseo de ratos submetidos à ausência de carga

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

Interações orais em língua inglesa no laboratório de multimídia com acesso à Internet

Pós-graduação em Estudos Linguísticos - IBILCE