Hypoxanthine-guanine phosphoribosyltransferase from Mycobacterium tuberculosis H37Rv: Cloning, expression, and biochemical characterization

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Functional shikimate dehydrogenase from Mycobacterium tuberculosis H37Rv: Purification and characterization

Tuberculosis (TB) poses a major worldwide public health problem. The increasing prevalence of TB, the emergence of multi-drug-resistant strains of Mycobacterium tuberculosis, the causative agent of TB, and the devastating effect of co-infection with HIV have highlighted the urgent need for the development of new antimycobacterial agents. Analysis of the complete genome sequence of M. tuberculosis shows the presence of genes involved in the aromatic amino acid biosynthetic pathway. Experimental evidence that this pathway is essential for M. tuberculosis has been reported. The genes and pathways that are essential for the growth of the microorganisms make them attractive drug targets since inhibiting their function may kill the bacilli. We have previously cloned and expressed in the soluble form the fourth shikimate pathway enzyme of the M. tuberculosis, the aroE-encoded shikimate dehydrogenase (mtSD). Here, we present the purification of active recombinant aroE-encoded M. tuberculosis shikimate dehydrogenase (mtSD) to homogeneity, N-terminal sequencing, mass spectrometry, assessment of the oligomeric state by gel filtration chromatography, determination of apparent steady-state kinetic parameters for both the forward and reverse directions, apparent equilibrium constant, thermal stability, and energy of activation for the enzyme-catalyzed chemical reaction. These results pave the way for structural and kinetic studies, which should aid in the rational design of mtSD inhibitors to be tested as antimycobacterial agents. (c) 2005 Elsevier B.V. All rights reserved.

Effect of soccer training on the running speed and the blood lactate concentration at the lactate minimum test

Tegtbur et al. [23] devised a new method able to estimate the intensity at maximal lactate steady state termed lactate minimum test. According to Billat et al. [7], no studies have yet been published on the affect of training on highest blood lactate concentration that can be maintained over time without continual blood lactate accumulation. Therefore, the aim of the present study was to verify the effect of soccer training on the running speed and the blood lactate concentration (BLC) at the lactate minimum test (Lac(min)). Thirteen Brazilian male professional soccer players, all members of the same team playing at National level, volunteered for this study. Measurements were carried out before (pre) and after (post) eight weeks of soccer training. The Lac(min) test was adapted to the procedures reported by Tegtbur et al. [23]. The running speed at the Lac(min) test was taken when the gradient of the line was zero. Differences in running speed and blood lactate concentration at the Lac(min) test before (pre) and after (post) the training program were evaluated by Student's paired t-test. The training program increased the running speed at the Lac(min) test (14.94 +/- 0.21 vs. 15.44 +/- 0.42* km(.)h(-1)) and the blood lactate concentration (5.11 +/- 2.31 vs. 6.93 +/- 1.33* mmol(.)L(-1)). The enhance in the blood lactate concentration may be explained by an increase in the lactate/H+ transport capacity of human skeletal muscle verified by other authors.

Máxima fase estável de lactato é ergômetro-dependente em modelo experimental utilizando ratos

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

Efeitos do treinamento de corrida em diferentes intensidades sobre a capacidade aeróbia e produção de lactato pelo músculo de ratos Wistar

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

Effects of Exercise Training on Hippocampus Concentrations of Insulin and IGF-1 in Diabetic Rats

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

Método para determinação de distribuição de tamanho de microbolhas (DTMB) em sistemas flotação (FAD) para tratamento de águas utilizando a análise de imagem digital

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

SAXS and UV-Vis combined to Quick-XAFS monitoring of ZnO nanoparticles formation and growth

Time-resolved X-ray absorption-fine structure (Quick-XAFS) and UV-Vis absorption spectroscopies were combined for monitoring simultaneously the time evolution of Zn-based species and ZnO quantum dot (Qdot) formation and growth during the sol-gel synthesis from zinc oxy-acetate precursor solution. The time evolution of the nanostructural features of colloidal suspension was independently monitored in situ by small angle X-ray scattering (SAXS). In both cases, the monitoring was initialized just after the addition of NaOH solution (B = [OH]/[Zn] = 0.5) to the precursor solution at 40 degrees C. Combined time-resolved Quick-XAFS and UV-Vis data showed that the formation of ZnO colloids from the zinc oxy-acetate consumption achieves a quasi-steady-state chemical equilibrium in less than 200s. Afterwards, the comparison of the ZnO Qdots size and Guinier gyration radius evidences a limited aggregation process coupled to the Qdots growth. The analysis of the experimental results demonstrates that the nanocrystal coalescence and Ostwald ripening control the kinetics of the Qdot growth.

Representações sobre meio ambiente de alunos da quarta série do Ensino Fundamental

Investigam-se as representações sociais sobre meio ambiente de 42 alunos de quartas séries do Ensino Fundamental em duas escolas públicas das zonas rural e urbana de um município do interior paulista. Os dados foram coletados mediante análise de documentos e pesquisa de campo, utilizando-se técnicas características da pesquisa qualitativa. Os resultados revelaram que, possivelmente, as origens daquelas representações - categorizadas como naturalistas e antropocêntricas estejam associadas, principalmente, às influências da mídia, família e religião. Acredita-se que os resultados possam contribuir para que os educadores considerem os saberes acumulados por seus alunos pela vivência de diferentes experiências e desenvolvam ações educativas ambientalmente comprometidas com a formação de indivíduos responsáveis pela criação e manutenção de melhores condições de vida, num contexto em que a prática pedagógica seja criativa, democrática e fundamentada no diálogo entre gerações e culturas, procurando estimular a ética nas relações entre os homens e entre estes e o meio ambiente.

Conservação de uvas Crimson Seedless e Itália, submetidas a diferentes tipos de embalagens e dióxido de enxofre (SO2)

Estudaram-se o efeito de diferentes tipos de embalagens e a ação do dióxido de enxofre (SO2) na conservação pós-colheita de uvas finas de mesa var. Crimson Seedless e Itália. Os frutos, colhidos em propriedade agrícola situada no município de Boa Vista-RR (Lat. 2º 50' 06 N e Long. 60º 40' 28 W), apresentavam, no momento da colheita, sólidos solúveis (SS) médios de 16,50 e 14,80°Brix, para as variedades Crimson Seedless e Itália, respectivamente. Antes da confecção dos tratamentos, os cachos foram higienizados em solução de hipoclorito de sódio (NaOCl) a 100 mg.L-1, previamente acidificada, por dez minutos. Utilizaram-se, para a atmosfera modificada passiva, sacolas de polietileno de baixa densidade (PEBD), sem perfuração, com 0,010; 0,015 e 0,020mm de espessura,e acondicionadas em embalagens secundárias de papelão (4kg) e de madeira (7,5kg). Para a geração do SO2, foram utilizados papéis Kraft de liberação rápida, com 3 e 8g de metabissulfito de sódio (Na2S2O5). Após a confecção dos tratamentos, os frutos foram armazenados em câmara frigorífica a 4 ± 1°C e 95 ± 3% de umidade relativa (U.R.). As avaliações foram realizadas no momento da colheita e, 7; 21; 35; 42 e 56 dias de armazenamento refrigerado, quanto à porcentagem de perda de massa fresca, taxa de desgrana e de bagas deterioradas, qualidade do engaço e teor de SS dos frutos. Após oito semanas, foi realizado teste de preferência para as duas variedades. Verificou-se, em ambas as variedades, que as uvas submetidas à ação do gerador de SO2 , contendo 3g de metabissulfito de sódio e acondicionamento em embalagens de PEBD de 0,020mm de espessura, independentemente do tipo de embalagem secundária, apresentaram a menor perda de massa fresca, menor taxa de desgrana e de bagas deterioradas, e melhor qualidade do engaço. Os resultados da análise sensorial concordaram com os resultados das análises físico-químicas. Não foram detectadas diferenças nos teores de SS entre os tratamentos, em ambas as variedades. (Apoio: Roraima Agrofrutas).

Modelo teórico e experimental da reciclagem do Carbono-13 em tecidos de mamíferos e aves

A diferença entre fontes alimentares da ordem de 14 , originárias de plantas com ciclos fotossintéticos Carbono-3 (C3) e Carbono-4 (C4) e seus subprodutos, abre novas perspectivas para o estudo do metabolismo do carbono em aves e animais de pequeno porte. Os autores propõem um modelo teórico e experimental capaz de exprimir os resultados de enriquecimento relativo, delta per mil (delta ) da razão 13C/12C versus tempo em diferentes tecidos. Utilizou-se a equação y(t) = (y0 -- q/k) e-kt + q/k onde, y(t) é a concentração isotópica no tempo desejado, y0 a concentração isotópica inicial existente no tecido, k é uma constante de troca isotópica com unidade 1/tempo, t é unidade de tempo e q é a taxa de entrada de metabólitos que contém carbono, com valores de delta /tempo. Para fígado de galinhas que tiveram a ração de ciclo fotossintético C4 substituída por dieta C3 obteve-se a equação delta13C = -24,74 + 12,37 e-0.237(nT), com meia-vida (T) de 2,9 dias. O patamar de equilíbrio de substituição do carbono foi alcançado em --24,48 , de modo que praticamente 98,4% do conteúdo isotópico do C4 no fígado foi substituído por C3 após 5,6 meias-vidas. O modelo foi adequado para determinar o tempo de reciclagem total ou parcial da concentração de carbono nos tecidos em estudo.

Study of the hydrolysis of TEOS-TMOS mixtures under ultrasound stimulation

Pure and mixed tetramethoxysilane (TMOS) and tetraethoxysilane (TEOS) were hydrolyzed at 35 degrees C, using oxalic acid as a catalyst and ultrasound stimulation. The hydrolysis reaction was carried out in a specially designed device, in which a heat flow steady state, between the ultrasound source and an external thermostatic bath, was maintained, in the absence of reactions. The exothermic hydrolysis causes a time dependent thermal peak. An induction time is apparent in pure TEOS before the hydrolysis peaks starts, which has been explained by the initial immiscibility gap of the TEOS-water system. The induction time was found to be approximately of the same magnitude as in the HCl catalyzed hydrolysis, in spite of the uncertainty accompanying the peak definition. No induction period is apparent in pure TMOS, so that the hydrolysis starts with its maximum rate. Two independent thermal peaks in the mixed TMOS-TEOS samples were found, both associated to the respective hydrolyses of the pure component. The induction time for the TEOS hydrolysis is decreased as more alcohol (and silanol) is produced in the earlier TMOS hydrolysis. This effect is explained by improvement of homogenization by alcohol.

Non-exhaustive test for aerobic capacity determination in swimming rats

The aim of this study was to describe a double-bout exercise test for non-exhaustive aerobic capacity determination in swimming rats. Adult rats were Submitted to 4 swimming tests at different intensities (4%, 6%, 7%, and 8% of body mass), with intervals of 48 h between them. Two exercise bouts of equal intensity lasting 5 min were performed, separated by 2 min with blood collection for lactate analysis. For each intensity, delta lactate was determined by subtracting lactate concentration at the end of the first effort from the lactate at the end of the second effort. Individual linear interpolation of delta lactate concentration enabled determination of a null delta, equivalent to the critical load (CL). Maxima) lactate steady state (MLSS) was also determined. The estimated CL was of 4.8% body mass and the MLSS was observed at 100% of CL, with blood lactate of 5.20 mmol/L. At 90%, blood lactate stabilized, with a progressive increase to 110% CL. These results offer a potential determination of aerobic capacity in swimming rats.

GC analysis of organotin compounds using pulsed flame photometric detection and conventional flame photometric detection

Organotin compounds, largely used as biocides in antifouling paints, are among the most toxic materials introduced into the aquatic environment. Sensitive analytical methods are thus required to characterize their occurrence in environmental and biological matrices. The comparison between two different photometric detectors in terms of analytical performance was carried out for the analysis of organotin compounds. A flame photometric detector (FPD) and a pulsed flame photometric detector (PFPD) were optimized. Their respective sensitivity, linearity range and selectivity were evaluated. Limits of detection obtained for a tributyltin compound (TBT) were 5.0 and 0.9 pg (as Sn) for the FPD and PFPD, respectively, using a 390 nm filter. The PFPD showed higher selectivity, besides reduced gas consumption in the flame, and is very attractive for organotin compound speciation in complex environmental matrices.

Transcriptional activation of MMP-13 by periodontal pathogenic LPS requires p38 MAP kinase

Matrix metal loprotease-13 (MMP-13) is induced by pro-inflammatory cytokines and increased expression is associated with a number of pathological conditions such as tumor metastasis, osteoarthritis, rheumatoid arthritis and periodontal diseases. MMP-13 gene regulation and the signal transduction pathways activated in response to bacterial LPS are largely unknown. In these studies, the role of the mitogen-activated protein kinase (MAPK) pathways in the regulation of MMP-13 induced by lipopolysaccharide was investigated. Lipopolysaccharide from Escherichia coli and Actinobacillus actinomycetemcomitans significantly (P < 0.05) increased MMP-13 steady-state mRNA (average of 27% and 46% increase, respectively) in murine periodontal ligament fibroblasts. MMP-13 mRNA induction was significantly reduced by inhibition of p38 MAP kinase. Immunoblot analysis indicated that p38 signaling was required for LPS-induced MMP-13 expression. Lipopolysaccharide induced proximal promoter reporter (-660/+32 mMMP-13) gene activity required p38 signaling. Collectively, these results indicate that lipopolysaccharide-induced murine MMP-13 is regulated by p38 signaling through a transcriptional mechanism.