A systematic approach to identify STRE-binding proteins of the gsn glycogen synthase gene promoter in Neurospora crassa

The gene encoding glycogen synthase in Neurospora crassa (gsn) is transcriptionally down-regulated when mycelium is exposed to a heat shock from 30 to 45 degrees C. The gsn promoter has one stress response element (STRE) motif that is specifically bound by heat shock activated nuclear proteins. In this work, we used biochemical approaches together with mass spectrometric analysis to identify the proteins that bind to the STRE motif and could participate in the gsn transcription regulation during heat shock. Crude nuclear extract of heat-shocked mycelium was prepared and fractionated by affinity chromatography. The fractions exhibiting DNA-binding activity were identified by electrophoretic mobility shift assay (EMSA) using as probe a DNA fragment containing the STRE motif DNA-protein binding activity was confirmed by Southwestern analysis. The molecular mass (MM) of proteins was estimated by fractionating the crude nuclear extract by SDS-PAGE followed by EMSA analysis of the proteins corresponding to different MM intervals. Binding activity was detected at the 30-50 MM kDa interval. Fractionation of the crude nuclear proteins by IEF followed by EMSA analysis led to the identification of two active fractions belonging to the pIs intervals 3.54-4.08 and 6.77-7.31. The proteins comprising the MM and pI intervals previously identified were excised from a 2-DE gel, and subjected to mass spectrometric analysis (MALDI-TOF/TOF) after tryptic digestion. The proteins were identified by search against the MIPS and MIT N. crassa databases and five promising candidates were identified. Their structural characteristics and putative roles in the gsn transcription regulation are discussed.

A Genome-wide Screen for Neurospora crassa Transcription Factors Regulating Glycogen Metabolism

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

Ambient pH Controls Glycogen Levels by Regulating Glycogen Synthase Gene Expression in Neurospora crassa. New Insights into the pH Signaling Pathway

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

Carbon nuclear magnetic resonance spectroscopic fingerprinting of commercial gasoline: Pattern-recognition analyses for screening quality control purposes

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

GC Fingerprints Coupled to Pattern-Recognition Multivariate SIMCA Chemometric Analysis for Brazilian Gasoline Quality Studies

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Screening Brazilian commercial gasoline quality by hydrogen nuclear magnetic resonance spectroscopic fingerprintings and pattern-recognition multivariate chemometric analysis

The identification of gasoline adulteration by organic solvents is not an easy task, because compounds that constitute the solvents are already in gasoline composition. In this work, the combination of Hydrogen Nuclear Magnetic Resonance ((1)H NMR) spectroscopic fingerprintings with pattern-recognition multivariate Soft Independent Modeling of Class Analogy (SIMCA) chemometric analysis provides an original and alternative approach to screening Brazilian commercial gasoline quality in a Monitoring Program for Quality Control of Automotive Fuels. SIMCA was performed on spectroscopic fingerprints to classify the quality of representative commercial gasoline samples selected by Hierarchical Cluster Analysis (HCA) and collected over a 6-month period from different gas stations in the São Paulo state, Brazil. Following optimized the (1)H NMR-SIMCA algorithm, it was possible to correctly classify 92.0% of commercial gasoline samples, which is considered acceptable. The chemometric method is recommended for routine applications in Quality-Control Monitoring Programs, since its measurements are fast and can be easily automated. Also, police laboratories could employ this method for rapid screening analysis to discourage adulteration practices. (C) 2010 Elsevier B.V. All rights reserved.

Screening Brazilian Commercial Gasoline Quality by ASTM D6733 GC and Pattern-Recognition Multivariate SIMCA Chemometric Analysis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

(1)H NMR Fingerprinting of Brazilian Commercial Gasoline: Pattern-Recognition Analyses for Origin Authentication Purposes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Reproductive pattern comparison of Uca thayeri Rathbun, 1900 and U. uruguayensis Nobili, 1901 (Crustacea, Decapoda, Ocypodidae)

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

Electrophoretic protein pattern and acid phosphatase activity in the midgut extracts of Apis mellifera L. (Hymenoptera: Apidae) during metamorphosis

Foram pesquisadas variações no padrão eletroforético das proteínas e da atividade da fosfatase ácida contidas em extratos do intestino médio de Apis mellifera L. durante o último estágio larval e pupação com a finalidade de estabelecer um paralelo entre os resultados e os eventos da metamorfose. Verificou-se maior variedade de bandas protéicas durante o estágio de pré-pupa e menor na pupa de olho marrom. A atividade da fosfatase ácida foi maior durante o último estágio larval e menor na pupa de olho branco. A maior variedade de bandas protéicas na pré-pupa coincide com a histólise do epitélio larval e reconstituição do epitélio pupal, enquanto a menor variabilidade na pupa de olho marrom coincide com o fim da diferenciação do intestino médio. A maior atividade fosfatásica no último estágio larval pode ocorrer em razão da sua função na histólise do epitélio.

Behaviour of dispersion indices in pattern detection of a population of angico, Anadenanthera peregrina (Leguminosae)

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

Color pattern variation in Trichomycterus iheringi (Eigenmann, 1917) (Siluriformes: Trichomycteridae) from rio Itatinga and rio Claro, São Paulo, Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Shell utilization pattern of the hermit crab Clibanarius vittatus (Crustacea, Anomura) in an estuary at São Vicente, State of São Paulo, Brazil

O objetivo do presente estudo foi avaliar a utilização de conchas de gastrópodes por Clibanarius vittatus (Bosc, 1802), na Praia dos Pescadores, em São Vicente (SP), Brasil. Foram realizadas amostras mensais no período de maio de 2001 a abril de 2003, na região intertidal durante a maré baixa. Todos os animais foram pesados e tiveram o comprimento de seu escudo cefalotorácico mensurados, suas conchas identificadas e medidas quanto ao comprimento (mm), abertura (mm) e peso (g). Capturamos 2.344 ermitões (644 machos, 1.594 fêmeas não-ovígeras, 45 fêmeas ovígeras e 61 em intersexo), utilizando 13 espécies de conchas de gastrópodes, das quais Stramonita haemastoma (Linnaeus, 1767), Cymatium parthenopeum (Von Salis, 1793) e Achatina fulica (Bowdich, 1822) representaram mais de 98% das conchas utilizadas. Obteve-se significativa diferença de tamanho entre os sexos, com machos e indivíduos em intersexo atingindo maior porte físico do que fêmeas. Este padrão influenciou fortemente na ocupação e utilização de conchas, principalmente em A. fulica, que foi a maior concha obtida por C. vittatus, utilizada somente por machos e indivíduos em intersexo. Cymatium parthenopeum foi a única espécie de concha em que se obtiveram elevados coeficientes de determinação para todas os parâmetros analisados. Stramonita haemastoma apresentou elevados e significativos coeficientes de determinação, indicando que esta espécie é o principal recurso de concha ocupada na Praia dos Pescadores.

Spatial pattern detection modeling of thrips (Thrips tabaci) on onion fields

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)