Characteristics of a Monoacylglycerol Lipase Isolated from Pseudomonas sp. LP7315 -Hydrolysis and Synthesis of Monoglycerides

A monoacylglycerol lipase (MGL) was purified from Pseudomonas sp. LP7315 by ammonium sulfate precipitation, anion-exchange chromatography, and preparative electrophoresis. The purified enzyme was homogeneous on an SDS-polyacrylamide gel with a molecular mass of 59 kDa. Its hydrolytic activity was confirmed to be specific for monoglycerides: the enzyme did not hydrolyze diand triglycerides. MGL was found to be stable even after l-h incubation at 65℃. The hydrolytic activity depended not only on temperature and pH but also on the type of monoglyceride used. MGL also catalyzed monoglyceride synthesis at 65℃ in a solvent-free two-phase system, in which fatty acid droplets were dispersed in the glycerol phase with a low water content. The synthetic reaction proceeded at a constant rate for approximately 24 h and reached an equilibrium after 48 h of reaction. The initial rate of the synthetic reaction depended on several factors: the type of fatty acid used as the substrate, the amounts of fatty acid and glycerol, and the concentration of MGL in the glycerol phase. To analyze the effects of these factors, a kinetic model was developed based on the assumption that the adsorption equilibrium of MGL molecules at the interface between the two phases is the rate-determining factor for the synthetic reaction. The model was found to yield a good approximation of the initial synthetic rate under various reaction conditions. The analysis suggests that the adsorption behavior of MGL onto the interface had a large effect on the initial rate of the monoglyceride synthesis

Application of a pharmacological transcriptome filter identifies a shortlist of mouse glucocorticoid receptor target genes associated with memory consolidation

Glucocorticoids regulate memory consolidation, facilitating long-term storage of relevant information to adequately respond to future stressors in similar conditions. This effect of glucocorticoids is well-established and is observed in multiple types of behaviour that depend on various brain regions. By and large, higher glucocorticoid levels strengthen event-related memory, while inhibition of glucocorticoid signalling impairs consolidation. The mechanism underlying this glucocorticoid effect remains unclear, but it likely involves the transcriptional effects of the glucocorticoid receptor (GR). We here used a powerful paradigm to investigate the transcriptional effects of GR in the dorsal hippocampus of mice after training in an auditory fear conditioning task, aiming to identify a shortlist of GR target genes associated to memory consolidation. Therefore, we utilized in an explorative study the properties of selective GR modulators (CORT108297 and CORT118335), alongside the endogenous agonist corticosterone and the classical GR antagonist RU486, to pinpoint GR-dependent transcriptional changes. First, we confirmed that glucocorticoids can modulate memory strength via GR activation. Subsequently, by assessing the specific effects of the available GR-ligands on memory strength, we established a pharmacological filter which we imposed on the hippocampal transcriptome data. This identified a manageable shortlist of eight genes by which glucocorticoids may modulate memory consolidation, warranting in-depth follow-up. Overall, we showcase the strength of the concept of pharmacological transcriptome filtering, which can be readily applied to other research topics with an established role of glucocorticoids

Anti-viral actions and viral dynamics in the early phase of three different regimens of interferon treatment for chronic hepatitis C: differences between the twice-daily administration of interferon-beta treatment and the combination therapy with interferon-alpha plus ribavirin.

To improve the efficacy of interferon (IFN) treatment for chronic hepatitis C, we have proposed the twice-daily administration of IFN-beta as a promising induction therapy. In this study, we demonstrated differences between the clearance of circulating HCV-RNA and the induction of anti-viral actions during the first 2 weeks of treatment. Nine patients with a high viral load and genotype 1b were randomly assigned to 3 groups: group A received 3MU of IFN-beta twice a day at intervals of 5 and 19 h; group B received 3MU of IFN-beta twice a day at intervals of 10 and 14 h; group C received 6MU of IFN-alpha once a day with ribavirin. The expression of OAS2, PKR, and MxA in peripheral blood mononuclear cells (PBMCs) were quantified by real-time polymerase chain reaction method. The viral clearance showed a bi-phasic pattern, and those in the second phase of groups A and B were significantly steeper than that of group C. The peak level of OAS2 during the first phase was correlated with the first phase decay. The MxA expression tended to be higher in group A and B than in group C. The expression of these 3 proteins tended to decrease at day 6 in group C, but increase in groups A and B. These might make differences in the viral decay during the second phase</p

COMPARAÇÃO DE CUSTO-BENEFÍCIO DE DOIS MÉTODOS DE EXTRAÇÃO DE AFLATOXINAS B1, G1, B2, G2 EM AMOSTRAS DE AÇÚCAR MASCAVO: Extração QuEChERS X Extração Líquido-Líquido

Alguns fungos, por serem elementos microbianos encontrados em todos os lugares, como água, ar e solo, produzem toxinas denominadas de micotoxinas, que podem ser encontradas em alimentos, algumas são capazes de gerar efeitos tóxicos quando consumidas por pessoas e animais. Existem diversas variedades de micotoxinas, as mais estudadas são as aflatoxinas B1, G1, B2, G2. Para a determinação das mesmas, são indispensáveis alguns processos no preparo da amostra que antecedem os métodos quantitativos e qualitativos, como a cromatografia em camada delgada (CCD). Antes da amostra ser submetida a CCD é fundamental a etapa de extração, porém, é utilizada uma grande quantidade de solventes e demanda elevada de tempo para o desenvolvimento de técnica como a do método líquido-líquido. No entanto, atualmente vem sendo aplicado um método, denominado QuEChERS que se caracteriza pela extração rápida, confiável, fácil, econômica, robusta e segura de analitos a partir de matrizes complexas, com menor uso de solventes. Nesse estudo duas distintas metodologias de extração foram comparadas em relação aos custos, tempo de desenvolvimento e quantidade de uso de solventes/reagentes para implantação de um novo método com etapa de extração mais favorável para a determinação de aflatoxinas em açúcar mascavo. Os resultados obtidos através desse estudo certificaram a eficiência e os benefícios da metodologia QuEChERS nas extrações, pois foi possível detectar melhor custo-benefício considerando a rapidez e o investimento necessário para a realização da técnica quando comparado com a extração Líquido-Líquido

Duct lavage cytology for the detection of breast cancer: report of a case

Although many modalities have been established to diagnose breast cancers, it is sometimes difficult to reveal nonpalpable cases. Duct lavage cytology was originally established to reveal groups at high risk for breast cancers by detecting metaplastic ductal cells. We report here a case where duct lavage was useful for revealing a small cancer that had been undetected by repeated bloody nipple discharge and cytological examinations. Duct lavage cytology may be of use in cases where nipple discharge of unknown origin persists

Application of a pharmacological transcriptome filter identifies a shortlist of mouse glucocorticoid receptor target genes associated with memory consolidation

Glucocorticoids regulate memory consolidation, facilitating long-term storage of relevant information to adequately respond to future stressors in similar conditions. This effect of glucocorticoids is well-established and is observed in multiple types of behaviour that depend on various brain regions. By and large, higher glucocorticoid levels strengthen event-related memory, while inhibition of glucocorticoid signalling impairs consolidation. The mechanism underlying this glucocorticoid effect remains unclear, but it likely involves the transcriptional effects of the glucocorticoid receptor (GR). We here used a powerful paradigm to investigate the transcriptional effects of GR in the dorsal hippocampus of mice after training in an auditory fear conditioning task, aiming to identify a shortlist of GR target genes associated to memory consolidation. Therefore, we utilized in an explorative study the properties of selective GR modulators (CORT108297 and CORT118335), alongside the endogenous agonist corticosterone and the classical GR antagonist RU486, to pinpoint GR-dependent transcriptional changes. First, we confirmed that glucocorticoids can modulate memory strength via GR activation. Subsequently, by assessing the specific effects of the available GR-ligands on memory strength, we established a pharmacological filter which we imposed on the hippocampal transcriptome data. This identified a manageable shortlist of eight genes by which glucocorticoids may modulate memory consolidation, warranting in-depth follow-up. Overall, we showcase the strength of the concept of pharmacological transcriptome filtering, which can be readily applied to other research topics with an established role of glucocorticoids.Metabolic health: pathophysiological trajectories and therap