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.

Cytogenetic and molecular analysis of the Pantaneiro cattle breed

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Qualidade do café natural e despolpado após secagem em terreiro e com altas temperaturas

Objetivou-se no presente trabalho verificar a qualidade do café natural e despolpado durante a secagem em terreiro e secagem com ar aquecido a 40º e 60ºC. O trabalho foi realizado no Departamento de Engenharia e no Pólo de Tecnologia em Pós-Colheita do Café da Universidade Federal de Lavras (UFLA). A colheita do café, cultivar Topázio, foi seletiva. Parte do café foi despolpado e outra parte processado de forma natural. Uma parcela de cada tipo de café foi conduzida para a secagem em terreiro e outra parcela para secagem com temperaturas de 40ºC e 60ºC. Para avaliação da qualidade, foram feitos teste de condutividade elétrica e lixiviação de potássio; determinação de acidez titulável total; teste de acidez graxa; açúcares totais e redutores. Os resultados obtidos permitem concluir que: o tempo para secagem é afetado pelos diferentes tipos de secagem e processamentos; a condutividade elétrica, a lixiviação de potássio, a acidez titulável total e a acidez graxa aumentam com a elevação da temperatura de secagem, independente do tipo de processamento; os açúcares redutores e os açúcares totais diminuem com o aumento da tem peratura de secagem independente do tipo de processamento; a secagem com temperatura de 60ºC afetou negativamente a qualidade do café.

FLAVOR-CHANGING NEUTRAL CURRENTS IN SU(3)L-CIRCLE-TIMES-U(1)Y MODELS

We consider flavor changing neutral current effects coming from the Z' exchange in 3-3-1 models. We show that the mass of this extra neutral vector boson may be less than 2 TeV and discuss the problem of quark family discrimination.

Novel wasp toxin discriminates between neuronal and cardiac sodium channels

Pompilidotoxins (PMTXs), derived from the venom of solitary wasp has been known to facilitate synaptic transmission in the lobster neuromuscular junction, and a recent further study from rat trigeminal neurons revealed that the toxin slows Na+ channel inactivation without modifying activation process. Here we report that beta -PMTX modifies rat brain type II Na+ channel alpha -subunit (rBII) expressed in human embryonic kidney cells but fails to act on the rat heart alpha -subunit (rH1) at similar concentrations. We constructed a series of chimeric mutants of rBII and rH1 Na+ channels and compared modification of the steady-state Na+ currents by beta -PMTX. We found that a difference in a single amino acid between Glu-1616 in rBII and Gln-1615 in rH1 at the extracellular loop of D4S3-S4 is crucial for the action of beta -PMTX. PMTXs, which are small peptides with 13 amino acids, would be a potential tool for exploring a new functional moiety of Na+ channels.

Vibrational spectroscopy of the seselin crystal

Seselin, C14H12O3, is a coumarin which crystallizes in a monoclinic structure P2(1)/b(C-2h(5)) with four molecules per unit cell. In a Fourier-transform Raman spectroscopic study performed at room temperature, several normal modes were observed. Vibrational wavenumber and wave vector calculations using density functional theory were compared with experiment, which allowed the assignment of a number of normal modes of the crystal. Temperature-dependent Raman spectra were recorded between 10 and 300 K. No anomalies were observed in the phonon spectra, indicating that the monoclinic structure remains stable. Copyright (c) 2007 John Wiley & Sons, Ltd.

Evidence of thermostable amylolytic activity from Rhizopus microsporus var. rhizopodiformis using wheat bran and corncob as alternative carbon source

Rhizopus microsporus var. rhizopodiformis produced high levels of alpha-amylase and glucoamylase under solid state fermentation, with several agricultural residues, such as wheat bran, cassava flour, sugar cane bagasse, rice straw, corncob and crushed corncob as carbon sources. These materials were humidified with distilled water, tap water, or saline solutions-Segato Rizzatti (SR), Khanna or Vogel. The best substrate for amylase production was wheat bran with SR saline solution (1:2 v/v). Amylolytic activity was still improved (14.3%) with a mixture of wheat bran, corncob, starch and SR saline solution (1:1:0.3:4.6 w/w/w/v). The optimized culture conditions were initial pH 5, at 45 degrees C during 6 days and relative humidity around 76%. The crude extract exhibited temperature and pH optima around 65 degrees C and 4-5, respectively. Amylase activity was fully stable for 1 h at temperatures up to 75 degrees C, and at pH values between 2.5 and 7.5.

A comparative study of different diets to optimize cultivation of Hyalella azteca in the laboratory

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

5-HT1A, but not 5-HT2 and 5-HT7, receptors in the nucleus raphe magnus modulate hypoxia-induced hyperpnoea

Aim: In the present study, we assessed the role of 5-hydroxytryptamine (5-HT) receptors (5-HT1A, 5-HT2 and 5-HT7) in the nucleus raphe magnus (NRM) on the ventilatory and thermoregulatory responses to hypoxia.Methods: To this end, pulmonary ventilation (V-E) and body temperature (T-b) of male Wistar rats were measured in conscious rats, before and after a 0.1 mu L microinjection of WAY-100635 (5-HT1A receptor antagonist, 3 mu g 0.1 mu L-1, 56 mM), ketanserin (5-HT2 receptor antagonist, 2 mu g 0.1 mu L-1, 36 mM) and SB269970 (5-HT7 receptor antagonist, 4 mu g 0.1 mu L-1, 103 mM) into the NRM, followed by 60 min of severe hypoxia exposure (7% O-2).Results: Intra-NMR microinjection of vehicle (control rats) or 5-HT antagonists did not affect V-E or T-b during normoxic conditions. Exposure of rats to 7% O-2 evoked a typical hypoxia-induced anapyrexia after vehicle microinjections, which was not affected by microinjection of WAY-100635, SB269970 or ketanserin. The hypoxia-induced hyperpnoea was not affected by SB269970 and ketanserin intra-NMR. However, the treatment with WAY-100635 intra-NRM attenuated the hypoxia-induced hyperpnoea.Conclusion: These data suggest that 5-HT acting on 5-HT1A receptors in the NRM increases the hypoxic ventilatory response.

Patients' Expectations Before and Evaluation After Dental Implant Therapy

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Evaluation of the Possibility of Removing Staining by Repolishing Composite Resins Submitted to Artificial Aging

This in vitro research verified the possibility of eliminating staining caused by coffee and red wine in five composite resins, after being submitted to thermal cycling. Thirty-six specimens were prepared and immersed in water at 37 degrees C for 24 hours. After polishing, specimen color was measured in a spectrophotometer Cintra 10 UV (Visible Spectrometer, GBC, Braeside, VIC, Australia). All specimens were submitted to thermal cycling at temperatures of 5 and 55 degrees C with a dwell time of 1 minute, for 1,000 cycles in a 75% ethanol/water solution. After thermal cycling, the specimens were immersed in water at 37 degrees C until 7 days had elapsed from the time the specimens were prepared. All specimens were then taken to the spectrophotometer for color measurement. The specimens were divided into three groups (N = 12): distilled water (control), coffee, and red wine. For the staining process to occur on only one surface, all the sides, except one, of the surfaces were isolated with white wax. The specimens were immersed in one of the solutions at 37 degrees C for 14 days. The specimens were dried and taken to the spectrophotometer for color measurement. After this, the specimens were submitted to 20 mu m wear three times, and the color was measured after each one of the wear procedures. Calculation of the color difference was made using CIEDE2000 formula. According to the methodology used in this research, it was concluded that the staining caused by coffee and red wine was superficial and one wear of 20 mu m was sufficient to remove the discoloration.

Reproductive biology in the primitive relic Angiosperm Drimys brasiliensis (Winteraceae)

From field observations on Drimys brasiliensis, principally in the Botucatu region of São Paulo State, Brazil, new data on the reproductive biology, the rhythm of growth, and the development of lateral cymose inflorescences, flowers and fruits are presented. Pollination accelerates the rate of flower-development for about 4-6 days. Pollination experiments show that D. brasiliensis is not self-sterile; because of mechanical devices the sticky pollen grains do not normally come into contact with the stigmata unless an animal pollen vector is involved. The pollinators are diurnal Coleoptera, Diptera and Thysanoptera which eat from the pollen, lick from the stigmatic exudates and (in case of the flies) probably also from the staminal glands. Fruit- and seedeaters are birds which seem to be the main dispersal agents. Establishment of new individuals normally is through seedlings, but also by vegetative propagation through plagiotropous branches which may root and separate from the mother plant. The morphological, developmental and reproductive aspects in D. brasiliensis are discussed in a wider context, compared with data from other Magnoliidae, and related to aspects of early Angiosperm evolution. © 1980 Springer-Verlag.

Effect of whitening agents on dentin bonding

Background: Several studies have shown a reduction in enamel bond strengths when the bonding procedure is carried out immediately after vital bleaching with peroxides. This reduction in bond strengths has become a concern in cosmetic dentistry with the introduction of new in-office and waiting-room bleaching techniques. The aim of this in vitro study was to evaluate the effect of three bleaching regimens: 35% hydrogen peroxide (HP), 35% carbamide peroxide (CP), and 10% CP, on dentin bond strengths. Materials and Methods: One hundred and twenty fresh bovine incisors were used in this study. The labial surface of each tooth was ground flat to expose dentin and was subsequently polished with 600-grit wet silicon carbide paper. The remaining dentin thickness was monitored and kept at an average of 2 mm. The teeth were randomly assigned to four bleaching regimens (n = 30): (A) control, no bleaching treatment; (B) 35% HP for 30 minutes; (C) 35% CP for 30 minutes; and (D) 10% CP for 6 hours. For each group, half of the specimens (n = 15) were bonded with Single Bond/Z100 immediately after the bleaching treatment, whereas the other half was bonded after the specimens were stored for 1 week in artificial saliva at 37°C. The specimens were fractured in shear using an Instron machine. Results: For the groups bonded immediately after bleaching, one-way analysis of variance (ANOVA) followed by the Duncan's post hoc test revealed a statistically significant reduction in bond strengths in a range from 71% to 76%. For the groups bonded at 1 week, one-way ANOVA showed that group B (35% HP for 30 min) resulted in the highest bond strengths, whereas 10% CP resulted in the lowest bond strengths. Student's t-test showed that delayed bonding resulted in a significant increase in bond strengths for groups B (35% HP) and C (35% CP); whereas the group bleached with 10% CP (group D) remained in the same range obtained for immediate bonding. Storage in artificial saliva also affected the control group, reducing its bond strengths to 53% of the original. ©2000 BC Decker Inc.

Effects of bleaching with carbamide peroxide gels on microhardness of restoration materials

The objective of this in vitro study was to quantitatively assess the effects of bleaching with 10 and 15% carbamide peroxide (CP) on restoration materials by performing superficial microhardness analysis. Acrylic cylindrical containers (4 x 2 mm) were filled with the following restoration products: Charisma (Heraues Kulzer, Vila Santa Catarina, São Paulo, Brazil), Durafill VS (Heraeus Kulzer), Vitremer (3M, Sumaré, São Paulo, Brazil), Dyract (Dentsply, Petrópolis, Rio de Janeiro, Brazil), and Permite C (SDI, São Pauio, São Paulo, Brazil). Sixty samples were prepared of each restoration material. Twenty samples received bleaching treatment with 10% CP, 20 samples received bleaching treatment with 15% CP, and 20 samples were kept submerged in artificial saliva, which was replaced daily. The treatment consisted of immersion of the specimens in 1 cm3 of CP at 10 and 15% for 6 hours per day during 3 weeks, whereupon the test specimens were washed, dried, and kept immersed in artificial saliva for 18 hours. Then the test and control specimens were analyzed using a microhardness gauge. The Knoop Hardness Number (KHN) was taken for each test and control specimen at five different locations by applying a 25 g force for 20 seconds. The values obtained were transformed into KHNs and the mean was calculated. The data were submitted to statistical analysis by analysis of variance and Tukey test, p < .05. The means/standard deviations were as follows: Charisma: CP 10% 38.52/4.08, CP 15% 34.31/6.13, saliva 37.36/4.48; Durafill VS: CP 10% 18.65/1.65, CP 15% 19.38/2.23, saliva 18.27/1.43; Dyract AP: CP 10% 30.26/2.81, CP 15% 28.64/5.44, saliva 33.88/3.46; Vitremer: CP 10% 28.15/3.04, CP 15% 17.40/3.11, saliva 40.93/4.18; and Permite C: CP 10% 183.50/27.09, CP 15% 159.45/5.78, saliva 215.80/26.15. A decrease in microhardness was observed for the materials Dyract AP, Vitremer, and Permite C after treatment with CP at 10 and 15%, whereas no effect on either of the two composites (Charisma and Durafill) was verified. CLINICAL SIGNIFICANCE: The application of the carbamide peroxide gels at 10 and 15% did not alter the microhardness of the composite resins Charisma and Durafill. In situ and clinical studies are necessary to enable one to conclude that the reduction in microhardness of the materials effectively results in clinical harm to the restorations.