Identification, characterization and expression analysis of hepcidin gene in sheep

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

NaF treatment increases TNF-alpha and resistin concentrations and reduces insulin signal in rats

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

Photoelastic analysis to compare implant-retained and conventional obturator dentures

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

Genome-Wide Analysis of Differentially Expressed Genes During the Early Stages of Tomato Infection by a Potyvirus

Plant responses against pathogens cause up-and downward shifts in gene expression. To identify differentially expressed genes in a plant-virus interaction, susceptible tomato plants were inoculated with the potyvirus Pepper yellow mosaic virus (PepYMV) and a subtractive library was constructed from inoculated leaves at 72 h after inoculation. Several genes were identified as upregulated, including genes involved in plant defense responses (e. g., pathogenesis-related protein 5), regulation of the cell cycle (e. g., cytokinin-repressed proteins), signal transduction (e. g., CAX-interacting protein 4, SNF1 kinase), transcriptional regulators (e. g., WRKY and SCARECROW transcription factors), stress response proteins (e. g., Hsp90, DNA-J, 20S proteasome alpha subunit B, translationally controlled tumor protein), ubiquitins (e. g., polyubiquitin, ubiquitin activating enzyme 2), among others. Downregulated genes were also identified, which likewise display identity with genes involved in several metabolic pathways. Differential expression of selected genes was validated by macroarray analysis and quantitative real-time polymerase chain reaction. The possible roles played by some of these genes in the viral infection cycle are discussed.

Molecular cytogenetic analysis of heterochromatin in the chromosomes of tilapia, Oreochromis niloticus (Teleostei : Cichlidae)

The structure of the heterochromatic bands in mitotic chromosomes of the important tropical aquaculture species of tilapia, Oreochromis niloticus, was investigated by the combination of the C-banding technique, chromosomal digestion with two restriction endonucleases and fluorescence in situ hybridization (FISH) of two satellite DNAs (SATA and SATB). The tilapia chromosomes presented heterochromatic bands in the centromeres and in the short arms of almost all chromosomes that were differentially digested by the restriction endonucleases HaeIII and EcoRI. FISH of SATA showed that the satellite sequence is distributed in the centromeric region of all chromosomes of tilapia. FISH also revealed an intense hybridization signal for SATB in only one chromosome pair, but less intense signals were also present in several other pairs. The digestion of tilapia chromosomes by HaeIII and EcoRI was positively correlated with the position of SATA and SATB in chromosomes as revealed by FISH. The results obtained may be useful in future molecular and genetic studies of tilapias.

Cytogenetic analysis of three species of the genus Haemulon (Teleostei : Haemulinae) from Margarita Island, Venezuela

This paper describes the karyotype analysis of Haemulon aurolineatum, Haemulon bonariensis and Haemulon plumierii, by Giemsa staining, C-banding, Ag-staining and fluorescent in situ hybridization (FISH), to locate the 18S and 5S rRNA genes. Diploid modal count in the three species was 2n = 48 acrocentric elements. Except for pair 24, which exhibited an unmistakable secondary constriction in all three species, it was not possible to classify them as homologous to each other because differences in chromosome size were too slight between adjacent pairs within a size-graded series. Ag-NOR clusters were located in pair 24 in the three species with signal located on the secondary constriction of these chromosomes. C-banding demonstrated that the three species share the same distribution pattern of the constitutive heterochromatin with centromeric heterochromatic blocks in the 23 chromosome pairs and a pericentromeric block in pair 24 which is coincident with the NORs. FISH experiments showed that 18S rDNA sequences were located coincident with the Ag-NOR site in the three species; however, differences in both the number and chromosome distribution of 5S-rDNA cluster were detected among them. Our data suggest that chromosome evolution of Haemulon has been preserved from major changes in the karyotypic macrostructure, whereas microstructural changes have occurred.

A solid paraffin-based carbon paste electrode modified with 2-aminothiazole organofunctionalized silica for differential pulse adsorptive stripping analysis of nickel in ethanol fuel

A solid paraffin-based carbon paste electrode modified with 2-aminothiazole organofunctionalized silica (SiAt-SPCPE) was applied to Ni2+ determination in commercial ethanol fuel samples. The proposed method comprised four steps: (1) Ni2+ preconcentration at open circuit potential directly in the ethanol fuel sample, (2) transference of the electrode to an electrochemical cell containing DMG, (3) differential pulse voltammogram registering and (4) surface regeneration by polishing the electrode. The proposed method combines the high Ni2+ adsorption capacity presented by 2-aminothiazole organofunctionalized silica with the electrochemical properties of the Ni(DMG)2 complex, whose electrochemical reduction provides the analytical signal.All experimental parameters involved in the proposed method were optimized. Using a preconcentration time of 20 min, it was obtained a linear range from 7.5 x 10(-9) to 1.0 x 10(-6) mol L-1 with detection limit of 2.0 x 10(-9) mol L-1. Recovery values between 96.5 and 102.4% were obtained for commercial samples spiked with 1.0 mu mol L-1 Ni2+ and the developed electrode was totally stable in ethanolic solutions. The contents of Ni2+ found in the commercial samples using the proposed method were compared to those obtained by graphite furnace atomic absorption spectroscopy by using the F- and t-test. Neither the F- nor t-values exceeded the critical values at 95% confidence level, confirming that there are not statistical differences between the results obtained by both methods. These results indicate that the developed electrode can be successfully employed to reliable Ni2+ determination in commercial ethanol fuel samples without any sample pretreatment or dilution step. (c) 2006 Elsevier B.V. All rights reserved.

Spectral analysis of the electromyograph of the erector spinae muscle before and after a dynamic manual load-lifting test

The aim of the present study was to assess the spectral behavior of the erector spinae muscle during isometric contractions performed before and after a dynamic manual load-lifting test carried out by the trunk in order to determine the capacity of muscle to perform this task. Nine healthy female students participated in the experiment. Their average age, height, and body mass (± SD) were 20 ± 1 years, 1.6 ± 0.03 m, and 53 ± 4 kg, respectively. The development of muscle fatigue was assessed by spectral analysis (median frequency) and root mean square with time. The test consisted of repeated bending movements from the trunk, starting from a 45º angle of flexion, with the application of approximately 15, 25 and 50% of maximum individual load, to the stand up position. The protocol used proved to be more reliable with loads exceeding 50% of the maximum for the identification of muscle fatigue by electromyography as a function of time. Most of the volunteers showed an increase in root mean square versus time on both the right (N = 7) and the left (N = 6) side, indicating a tendency to become fatigued. With respect to the changes in median frequency of the electromyographic signal, the loads used in this study had no significant effect on either the right or the left side of the erector spinae muscle at this frequency, suggesting that a higher amount and percentage of loads would produce more substantial results in the study of isotonic contractions.

Back extensor muscle fatigue at submaximal workloads assessed using frequency banding of the electromyographic signal

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

Crystallization and preliminary crystallographic analysis of SMase I, a sphingomyelinase from Loxosceles laeta spider venom

SMase I, a 32 kDa sphingomyelinase found in Loxosceles laeta venom, is responsible for the major pathological effects of spider envenomation. This toxin has been cloned and functionally expressed as a fusion protein containing a 6 x His tag at its N-terminus to yield a 33 kDa protein [Fernandes-Pedrosa et al. (2002), Biochem. Biophys. Res. Commun. 298, 638 - 645]. The recombinant protein possesses all the biological properties ascribed to the whole L. laeta venom, including dermonecrotic and complement-dependent haemolytic activities. Dynamic light-scattering experiments conducted at 291 K demonstrate that the sample possesses a monomodal distribution, with a hydrodynamic radius of 3.57 nm. L. laeta SMase I was crystallized by the hanging-drop vapour-diffusion technique using the sparse-matrix method. Single crystals were obtained using a buffer solution consisting of 0.08 M HEPES and 0.9 M trisodium citrate, which was titrated to pH 7.5 using 0.25 M sodium hydroxide. Complete three-dimensional diffraction data were collected to 1.8 Angstrom at the Laboratorio Nacional de Luz Sincrotron (LNLS, Campinas, Brazil). The crystals belong to the hexagonal system ( space group P6(1) or P6(5)), with unit-cell parameters a = b = 140.6, c = 113.6 Angstrom. A search for heavy-atom derivatives has been initiated and elucidation of the crystal structure is currently in progress.

Crystallization and preliminary X-ray diffraction analysis of a class II phospholipase D from Loxosceles intermedia venom

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

Diagnosis of inflammatory lesions by high-wavenumber FT-Raman spectroscopy

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

Search for light resonances decaying into pairs of muons as a signal of new physics

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Caracterização do perfil comunicativo de crianças com distúrbios do espectro autístico com diferentes interlocutores

OBJETIVO: caracterizar e comparar o perfil comunicativo de crianças com distúrbios do espectro autístico com dois diferentes interlocutores. MÉTODO: participaram da pesquisa nove sujeitos, sendo quatro crianças do gênero masculino, com faixa etária variando entre cinco anos e oito meses e onze anos, com distúrbios do espectro autístico e cinco terapeutas de linguagem. Foram realizadas análises do perfil comunicativo dessas crianças com os dois diferentes interlocutores (terapeuta de linguagem da própria criança e terapeuta desconhecida). Para análise dos dados foi empregado o Protocolo de Pragmática e tratamento estatístico dos resultados (p < 0,05; Teste dos Postos Sinalizados de Wilcoxon). RESULTADOS: foram levantados o número de atos comunicativos, o meio pelo qual esses atos foram expressos (vocal, verbal e gestual) e as funções comunicativas. em média, o número de atos comunicativos expressos pelos sujeitos tanto com os terapeutas das próprias crianças (5,10 atos/min) como com a terapeuta desconhecida (4,93 atos/min) não foi estatisticamente significante. Observou-se na situação de interação com o próprio terapeuta que a média do percentual da ocorrência do meio comunicativo verbal (31,50%) e a média de funções mais interativas (33,25%) são maiores do que com terapeuta desconhecida. CONCLUSÃO: os diferentes interlocutores, estudados nesta pesquisa, não tiveram uma influência no perfil comunicativo destas crianças. Deste modo, sugerem-se novos estudos, com inclusão de outros interlocutores, a fim de estabelecer quais variáveis interacionais interferem estatisticamente no perfil comunicativo de crianças do espectro autístico, com o intuito de contribuir para a elaboração de melhores propostas terapêuticas para esta população.

Adsorptive potentiometric stripping analysis of nucleic acids at mercury electrodes

The application of adsorptive stripping potentiometry to the reductive detection of nucleic acids at mercury electrodes is reported. Compared to analogous voltammetric stripping modes, constant current potentiometric stripping analysis (PSA) effectively addresses the hydrogen discharge background problem, and hence greatly improves the characteristics of the superimposed cytosine/adenine (CA) reduction peak. Compared to earlier schemes for trace measurements of nucleic acids at mercury or carbon electrodes that rely on anodic signals arising from the guanine residue, convenient quantitation can now be carried out in connection with the cytosine and adenine residues. Variables influencing the adsorptive PSA response are explored and optimized. With five minute accumulation, the detection limits for tRNA, ssDNA and dsDNA are 30 mu g l(-1), 60 mu g l(-1) and 2 mg l(-1), respectively. Such different values reflect the strong dependence of the PSA CA signal upon the nucleic-acid structure. This allows the quantitation of ssDNA or tRNA in the presence of dsDNA, and offers new possibilities for electrochemical studies of DNA structure and interactions.