Carbohydrate metabolism of Xylella fastidiosa: Detection of glycolytic and pentose phosphate pathway enzymes and cloning and expression of the enolase gene

The objective of this work was to assess the functionality of the glycolytic pathways in the bacterium Xylella fastidiosa. To this effect, the enzymes phosphoglucose isomerase, aldolase, glyceraldehyde-3-phosphate dehydrogenase and pyruvate kinase of the glycolytic pathway, and glucose 6-phosphate dehydrogenase of the Entner-Doudoroff pathway were studied, followed by cloning and expression studies of the enolase gene and determination of its activity. These studies showed that X. fastidiosa does not use the glycolytic pathway to metabolize carbohydrates, which explains the increased duplication time of this phytopatogen. Recombinant enolase was expressed as inclusion bodies and solubilized with urea (most efficient extractor), Triton X-100, and TCA. Enolase extracted from X. fastidiosa and from chicken muscle and liver is irreversibly inactivated by urea. The purification of enolase was partial and resulted in a low yield. No enzymatic activity was detected for either recombinant and native enolases, aldolase, and glyceraldehyde-3-phosphate dehydrogenase, suggesting that X. fastidiosa uses the Entner-Doudoroff pathway to produce pyruvate. Evidence is presented supporting the idea that the regulation of genes and the presence of isoforms with regulation patterns might make it difficult to understand the metabolism of carbohydrates in X. fastidiosa.

Enolase from Paracoccidioides brasiliensis: isolation and identification as a fibronectin-binding protein

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

Surface-expressed enolase contributes to the adhesion of Paracoccidioides brasiliensis to host cells

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo de isoenzimas de metabolismo de carboidratos e clonagem e expressão da enolase de Xylella fastidiosa

Pós-graduação em Agronomia (Genética e Melhoramento de Plantas) - FCAV

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

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Comparative transcriptome analysis of Paracoccidioides brasiliensis during in vitro adhesion to type I collagen and fibronectin: identification of potential adhesins

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Expressão antigênica em carcinomas do plexo coróide humano: relato de dois casos

Neoplasias provenientes do epitélio de revestimento do plexo coróide são inco-muns, tendo sido descritos 6 padrões morfológicos. O padrão anaplásico, também denominado carcinoma do plexo coróide, é o de menor freqüência e pode dar metastases fora do SNC. A distinção histológica desses tumores, particularmente da variedade anaplásica, com outras neoplasias primárias e metastáticas no SNC pode ser difícil. O uso de técnicas imunocitoquimicas em parafina tem-se mostrado útil no esclarecimento das linhagens tumorais. Os papilomas do plexo coróide têm, no entanto, sido objeto de controvérsia, por sua complexa expressão antigênica. Usando a técnica de imunoperoxidase (sistema avidina-biotina-peroxidase) pesquisaram-se, em dois casos da variedade anaplásica, os seguintes marcadores: proteína glial fibrilar ácida (GFAP) com anticorpo monoclonal e policlonal; ceratinas de 40-50kDa, ceratinas de 60-70kDa (callus ceratina), enolase neuronal específica (NSE) e proteína S-100, com anticorpos monoclonais. Os dois tumores mostraram positividade para NSE, proteína S-100 e ceratina de 40-50kDa; uma das duas neoplasias mostrou diferenciação glial, revelando positividade para GFAP tanto com anticorpo monoclonal quanto policlonal.

Carcinoid tumour of male breast diagnosed by fine needle aspiration.

A primary carcinoid tumour of the breast in a 66-year-old man was diagnosed by fine-needle aspiration cytology. The nature of the lesion was proved by histochemical and immunocytochemical studies. The importance of a conclusive diagnosis is discussed and the value of immunocytochemical analysis as an aid to cytomorphologic diagnosis is demonstrated.

Adenocarcinoma de cólon com diferenciação neuro-endócrina e escamosa. Relato de caso.

A tubular adenocarcinoma of the colon with solid area composed by small cells that was found by immunohistochemistry study using antibody to neuron-specific enolase (NSE) to possess neuroendocrine differentiation is related. In another areas of the tumor were visualized keratinizing squamous cells. The presence of neuro-endocrine and squamous cells features provide further evidence that neoplastic colonic cells have the capacity for multi-directional differentiation. The implications of this combination in relation to theories of tumor origin and differentiation and the prognostic significance of neuro-endocrine cells in malignant neoplasms of the gastrointestinal tract are discussed.

Expressão gênica das adesinas e fatores de virulência da fase miceliar e leveduriforme do complexo paracoccidioides e de sua interação com células epiteliais e fagócitos

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Investigação de marcadores protéicos do carcinoma oral

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influência da amoxicilina na virulência do Helicobacter pylori

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização e identificação de moléculas de superfície presentes em Paracoccidioides spp

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)

Alterations of protein expression in conditions of copper-deprivation for Paracoccidioides lutzii in the presence of extracellular matrix components

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