Characterization of the interdependency between residues that bind the substrate in a β-glycosidase

The manner by which effects of simultaneous mutations combine to change enzymatic activity is not easily predictable because these effects are not always additive in a linear manner. Hence, the characterization of the effects of simultaneous mutations of amino acid residues that bind the substrate can make a significant contribution to the understanding of the substrate specificity of enzymes. In the β-glycosidase from Spodoptera frugiperda (Sfβgly), both residues Q39 and E451 interact with the substrate and this is essential for defining substrate specificity. Double mutants of Sfβgly (A451E39, S451E39 and S451N39) were prepared by site-directed mutagenesis, expressed in bacteria and purified using affinity chromatography. These enzymes were characterized using p-nitrophenyl β-galactoside and p-nitrophenyl β-fucoside as substrates. The k cat/Km ratio for single and double mutants of Sfβgly containing site-directed mutations at positions Q39 and E451 was used to demonstrate that the effect on the free energy of ES‡ (enzyme-transition state complex) of the double mutations (∆∆G‡xy) is not the sum of the effects resulting from the single mutations (∆∆G‡x and ∆∆G‡y). This difference in ∆∆G‡ indicates that the effects of the single mutations partially overlap. Hence, this common effect counts only once in ∆∆G‡xy. Crystallographic data on β-glycosidases reveal the presence of a bidentate hydrogen bond involving residues Q39 and E451 and the same hydroxyl group of the substrate. Therefore, both thermodynamic and crystallographic data suggest that residues Q39 and E451 exert a mutual influence on their respective interactions with the substrate.FAPESPCNP

Estado Da Arte Da Literatura Científica Sobre Hancornia Speciosa: Tendências E Lacunas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)The economic and scientific interest in Hancornia speciosa (mangabeira) has been growing in recent years, mainly due to the marketing of fruit and extraction of natural compounds with high pharmacological potential. In this study, a scientometric survey about mangabeira was carried out in order to promote and direct future studies on the species. As a result, low scientific productivity associated with this species was found, with only 131 articles published in the last 69 years. In addition, this study identified some trends in bibliographic production on mangabeira, among them: the increasing number of articles over the years; scientific dissemination in nationwide journals; the main focus of this study is associated with agronomic studies; the experimental approach is more frequent and usually associated with populations of restricted geographical distribution; and the scientific production is mainly from education institutions. Furthermore, this study also allowed the identification of some gaps in knowledge about mangabeira, among them the difficulty in describing and characterizing botanical lines; lack of analysis of the genetic diversity of widely distributed populations; lack of management and conservation projects for the species; lack of description of cultivation, collection and preservation techniques of fruits; and lack of identification of natural compounds responsible for its pharmacological activity. It is expected that the data generated in this study will serve to direct future studies on H. speciosa. © 2016, Sociedade Brasileira de Fruticultura. All rights reserved.384CAPES, Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorCNPq, Conselho Nacional de Desenvolvimento Científico e TecnológicoFAPEG, Fundação de Amparo à Pesquisa do Estado de GoiásFINEP, Financiadora de Estudos e ProjetosCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Modelling the kinetics of peroxidase inactivation, colour and texture changes of pumpkin (Cucurbita maxima L.) during blanching

The effects of blanching treatment on peroxidase inactivation, colour and texture of pumpkin (Cucurbita maxima L.) were studied in the temperature range of 75–95 C. Peroxidase inactivation followed a first-order Arrhenius model, where the activation energy and rate of the reaction at a reference temperature of 85 C were 86.20 ± 5.57 kJ mol 1 and 0.27 ± 0.01 min 1, respectively. During blanching, pumpkin became darker and softer with processing time. The degradation of colour (evaluated throughout CIE L*a*b* colour system, with chroma index and total colour difference) and texture parameters (firmness and energy) showed a fractional conversion model kinetics, being the temperature effect on kinetic parameters well described by the Arrhenius law. The results of this work are a good tool to further optimise pumpkin blanching conditions

Echolucency of carotid plaques correlates with plaque cellularity

AbstractObjective: to analyse the relationship between carotid plaque echolucency and cellularity. Methods: carotid plaques (14 symptomatic and 16 asymptomatic) were snap frozen after endarterectomy and defined on the basis of their grey-scale-median (GSM), obtained from pre-operative high-definition ultrasonography, as either echolucent (<32) or echogenic (≥32). DNA and total soluble protein were determined to assess cellularity. Results: after correcting for wet weight, symptomatic plaques had significantly more DNA (0.400 ± 0.07 vs 0.335 ± 0.07 mg/g; p = 0.03) and soluble protein (34.1 ± 6.6 vs 29.7 ± 3.4 mg/g; p = 0.03) than asymptomatic plaques. Predominantly echolucent (Grey-Weale classification) plaques had more DNA (0.404 ± 0.06 vs 0.332 ± 0.08 mg/g; p = 0.03) than echogenic plaques. Plaques with GSM < 32 also had more DNA (0.386 ± 0.08 vs 0.319 ± 0.06 mg/g; p = 0.04) and soluble protein (34.7 ± 7.3 vs 29.6 ± 4.2 mg/g; p = 0.03) than those with GSM ≥ 32. Inverse relations were found between GSM and plaque DNA (r = −0.47; p = 0.02) and soluble protein (r = −0.45; p = 0.02) as well as between age and DNA (r = 0.39; p = 0.04) and soluble protein (r = −0.50; p = 0.003). Conclusions: echolucency of carotid plaques as assessed by ultrasonography reflects plaque cellularity. This observation support the notion that ultrasonography can be used to identify high-risk plaques and evaluate effect of interventions on plaque structure.Eur J Vasc Endovasc Surg 26, 32-38 (2003

Activity of daptomycin- and vancomycin-loaded poly-epsilon-caprolactone microparticles against mature staphylococcal biofilms.

The aim of the present study was to develop novel daptomycin-loaded poly-epsilon-caprolactone (PCL) microparticles with enhanced antibiofilm activity against mature biofilms of clinically relevant bacteria, methicillin-resistant Staphylococcus aureus (MRSA) and polysaccharide intercellular adhesin-positive Staphylococcus epidermidis. Daptomycin was encapsulated into PCL microparticles by a double emulsion-solvent evaporation method. For comparison purposes, formulations containing vancomycin were also prepared. Particle morphology, size distribution, encapsulation efficiency, surface charge, thermal behavior, and in vitro release were assessed. All formulations exhibited a spherical morphology, micrometer size, and negative surface charge. From a very early time stage, the released concentrations of daptomycin and vancomycin were higher than the minimal inhibitory concentration and continued so up to 72 hours. Daptomycin presented a sustained release profile with increasing concentrations of the drug being released up to 72 hours, whereas the release of vancomycin stabilized at 24 hours. The antibacterial activity of the microparticles was assessed by isothermal microcalorimetry against planktonic and sessile MRSA and S. epidermidis. Regarding planktonic bacteria, daptomycin-loaded PCL microparticles presented the highest antibacterial activity against both strains. Isothermal microcalorimetry also revealed that lower concentrations of daptomycin-loaded microparticles were required to completely inhibit the recovery of mature MRSA and S. epidermidis biofilms. Further characterization of the effect of daptomycin-loaded PCL microparticles on mature biofilms was performed by fluorescence in situ hybridization. Fluorescence in situ hybridization showed an important reduction in MRSA biofilm, whereas S. epidermidis biofilms, although inhibited, were not eradicated. In addition, an important attachment of the microparticles to MRSA and S. epidermidis biofilms was observed. Finally, all formulations proved to be biocompatible with both ISO compliant L929 fibroblasts and human MG63 osteoblast-like cells

Semigroup Closures of Finite Rank Symmetric Inverse Semigroups

We introduce the notion of semigroup with a tight ideal series and investigate their closures in semitopological semigroups, particularly inverse semigroups with continuous inversion. As a corollary we show that the symmetric inverse semigroup of finite transformations $\mathscr{I}_\lambda^n$ of the rank $\leqslant n$ is algebraically closed in the class of (semi)topological inverse semigroups with continuous inversion. We also derive related results about the nonexistence of (partial) compactifications of classes of semigroups that we consider.Comment: With the participation of the new coauthor - Jimmie Lawson - the manuscript has been substantially revised and expanded. Accordingly, we have also changed the manuscript titl

Avaliação e otimização da termossonicação como tratamento alternativo de pós-colheita de tomate fresco (Lycopersicum esculentum, cv. Zinac)

info:eu-repo/semantics/publishedVersio

Chemical Quality of Rainwater and Surface Runoff Water in the Proximity of the Abstractions That Supply the São Pedro do Sul Medical Spa (Portugal)

São Pedro do Sul medical spa provides health services using hot natural groundwater superiorly classified as natural mineral water. The main source of supply of that medical spa, has been over time, the Traditional Spring, which in recent decades has been systematically controlled, and shows spectacular constancy in its chemical quality, if there are no outside contaminations. Within the context presented, the detailed physical-chemical quality of rainwater and surface runoff water was studied, in the proximity of the Traditional Spring, as there is a potential for them to infiltrate at depth and evolve into the natural mineral water aquifer system. Thus, in the present chapter, after presenting the physical-chemical quality of the natural mineral water from the Traditional Spring, as well as some elements of the literature on the physical-chemical quality of rainwater, the methodology of work is followed, and then the results obtained from the physical-chemical composition of rainwater and surface run-off water are presented and discussed, comparing them with the quality of the water from the Traditional Spring. Finally, the main conclusions are presented, and some recommendations are made on research into potential sources of pollution, which justify the poor quality of the rainwater studied