Expanding the pH range of glutamate decarboxylase from L. pltarum LC84 by site-directed mutagenesis

Introduction: Glutamate decarboxylase is a class Ⅱ amino acid decarboxylase dependent onpyridoxal-5′-phosphate (PLP), which catalyzes the decarboxylation of substrateL-glutamate (L-Glu) to synthesize γ-aminobutyric acid (GABA). The low activity ofglutamic acid decarboxylase (GAD) and its ability to catalyze only under acidicconditions limit its application in biosynthesis of GABA.Methods: Taking glutamic acid decarboxylase from Lactobacillus plantarum, which produces GABA, as the research object, the mutation site was determined by amino acid sequence analysis of GAD, the mutation was introduced by primers, and the mutant was constructed by whole plasmid PCR and expressed in Escherichia coli. Then, the enzymatic properties of the mutant were analyzed. Finally, the three-dimensional structure of the mutant was simulated to support the experimental results.Results and Discussion: In this case, mutants E313S and Q347H of glutamate decarboxylase from L. pltarum LC84 (LpGAD) were constructed by targeted mutagenesis. Compared with the wild-type, their enzyme activity increased by 62.4% and 12.0% at the optimum pH 4.8, respectively. In the range of pH 4.0–7.0, their enzyme activity was higher than that of the wild-type, and enzyme activity of mutant E313S was 5 times that of the wild-type at pH 6.2. Visualization software PyMOL analyzed the 3D structure of the mutant predicted by homologous modeling, and the results showed that mutant E313S may broadened the reaction pH of LpGAD through the influence of surface charge, while mutant Q347H may broadened the reaction pH of LpGAD through the stacking effect of aromatic rings. In a word, mutants E313S and Q347H were improved the enzyme activity and were broadened the reaction pH of the enzyme, which made it possible for it to be applied in food industry and laid the foundation for the industrial production of GABA

The role of perceived severity of disaster, rumination, and trait resilience in the relationship between rainstorm-related experiences and PTSD amongst Chinese adolescents following rainstorm disasters

This study examined the mediating role of perceived severity of trauma and rumination in the relationship between traumatic experiences and PTSD, and assessed the moderated role of resilience in this mediating process. Nine hundred and fifty-one adolescents were selected to complete a self-report questionnaire involving rainstorm-related experiences, perceived severity of trauma, rumination, resilience, and PTSD. The results found that rainstorm-related experiences had a positive effect on PTSD by perceived severity of disaster, or by rumination via perceived severity of disaster. Resilience buffered the relationship between rainstorm-related experiences and PTSD, but did not buffer the relation of rainstorm-related experiences to perceived severity of disaster and rumination. These findings indicated that rainstorm-related experiences may have an indirect effect on PTSD via cognitive activities, and these indirect paths were not buffered by resilience. A buffering effect only occurred in the direct paths from rainstorm-related experiences to PTSD

Association of Toll-Like Receptor 4 Gene Polymorphism and Expression with Urinary Tract Infection Types in Adults

Background: Innate immunity of which Toll-like receptor (TLR) 4 and CXCR1 are key elements plays a central role in the development of urinary tract infection (UTI). Although the relation between the genetics of TLR4 and CXCR1 and UTI is investigated partly, the polymorphisms and expression of TLR4 and CXCR1 in different types of UTI in adults are not extremely clear. Methodology/Principal Findings: This study investigates the presence of TLR4 A (896) G and CXCR1 G (2608) C polymorphisms in 129 UTI patients using RFLP-PCR. Gene and allelic prevalence were compared with 248 healthy controls. Flow cytometry was used to detect TLR4 and CXCR1 expression in the monocytes of UTI patients and healthy controls. TLR4 (896) AG genotype and TLR4 (896) G allele had higher prevalence in UTI (especially in acute cystitis and urethritis) patients, whereas CXCR1 (2608) GC genotype and CXCR1 (2608) C allele had lower prevalence in UTI patients than controls. TLR4 expression was significantly lower in chronic UTI patients than in acute pyelonephritis or healthy controls. CXCR1 expression was similar in both controls and patients. TLR4 expression in chronic UTI patients after astragalus treatment was higher than pre-treatment. Conclusions: The results indicate the relationship between the carrier status of TLR4 (896) G alleles and the development of UTI, especially acute cystitis and urethritis, in adults. TLR4 expression levels are correlated with chronic UTI

Correlation and combining ability analysis of physiological traits and some agronomic traits in maize

Combining ability information on the physiological traits in maize (Zea mays L) and the relationship between physi¬ological traits and biomass, grain yield (GY) and yield components (YC) can help maize breeders design experi¬ments for improving inbred lines and/or developing hybrids with improved GY or YC (GYYC). A six-parent diallel experiment (Griffing method 3) was conducted for combining ability and correlation analyses. The objectives of this study were to 1) study the correlation between physiological traits and biomass at seedling stage; 2) study which physiological traits at seedling stage have significant correlation with biomasses at both seedling and later growth stages and GYYCs; 3) evaluate combining ability of the physiological traits that are significantly correlated with either GY or one of the YCs. Results showed plant heights at 20 day, 40 day, and leaf area were highly corre¬lated with both dry weights of shoots and roots. All chlorophyll-related organelles were significantly correlated with only dry weights of shoots. However, dry matter at seedling stage seemed not to be related to dry matter in later growth stages. Five physiological traits (stomatal conductance, transpiration rate, net photosynthetic rate, two quantum yield related traits) at seedling stage were identified to greatly impact dry matter at later growth stages. Results also showed that 13 out of 35 physiological traits studied were significantly correlated with GYYCs. Differ¬ent germplasms for improving GYYCs could be used based on both correlation between the 13 traits and GYYCs and combining ability effects of each line for the 13 selected traits

The impact of cognitive training on cerebral white matter in community-dwelling elderly : one-year prospective longitudinal diffusion tensor imaging study

It has been shown that cognitive training (CogTr) is effective and recuperative for older adults, and can be used to fight against cognitive decline. In this study, we investigated whether behavioural gains from CogTr would extend to white matter (WM) microstructure, and whether training-induced changes in WM integrity would be associated with improvements in cognitive function, using diffusion tensor imaging (DTI). 48 healthy community elderly were either assigned to multi-domain or single-domain CogTr groups to receive 24 sessions over 12 weeks, or to a control group. DTI was performed at both baseline and 12-month follow-up. Positive effects of multi-domain CogTr on long-term changes in DTI indices were found in posterior parietal WM. Participants in the multi-domain group showed a trend of long-term decrease in axial diffusivity (AD) without significant change in fractional anisotropy (FA), mean diffusivity (MD) or radial diffusivity (RD), while those in the control group displayed a significant FA decrease, and an increase in MD and RD. In addition, significant relationships between an improvement in processing speed and changes in RD, MD and AD were found in the multi-domain group. These findings support the hypothesis that plasticity of WM can be modified by CogTr, even in late adulthood

Sol-gel Synthesis of TiO2 With p-Type Response to Hydrogen Gas at Elevated Temperature

Titanium dioxide is considered as one of the potential candidates for high-temperature gas sensing applications due to its excellent sensitivity and stability. However, its practical use as a gas sensor under elevated conditions is limited on account of its selectivity and insufficient understanding of response conversion from n- to p-type. To this context, the present work is intended to prepare and understand the p-type response of anatase TiO2 toward H2 gas (20–1,000 ppm) at elevated temperature (500°C). Sol-gel route is adopted to facilely synthesize powders containing pure and chromium (1–10 at.%) doped TiO2 nanoparticles, which are then brushed onto substrates with already patterned inter-digitated platinum electrodes. In this work, even, the undoped TiO2 samples showed p-type gas sensing response, which then decreased with Cr doping. However, in comparison to previously reported work, the sensing characteristics of all sensors is improved. For instance, 5 at.% Cr-TiO2 showed high response (147), fast response and recovery (142/123s) time, and good selectivity to hydrogen against monoxide and methane. Despite better response values, the TiO2 based samples show instability and drift in baseline resistance; such issues were not observed for Cr-doped TiO2 samples (≥3 at.%). The powders were further analyzed by XRD, SEM, TEM, and XPS to understand the basic characteristics, p-type response and stability. Further, a plausible sensing mechanism is discussed on basis of results obtained from aforementioned techniques

Calculating the shear strength of a rock mass joint surface considering cyclic shear deterioration

The dynamic response of rock mass is largely restricted by its joint surface. Previous studies have shown that the degradation of joint surface can not be ignored when calculating the shear strength of structural plane under cyclic load. Although several studies have attempted to calculate the cyclic shear strength of a rock mass joint surface, an established and reliable method for calculating the cyclic shear strength of rock mass discontinuities is still lacking, thus necessitating further research. In this study, the deterioration effect of the shear strength of the joint surface under cyclic shearing was first analysed using cyclic shearing tests. The influence of vibration degradation in rock mass on the structural surface, undulant angle equation of the joint surface, and calculation method for the basic friction angle under a cyclic shearing load are proposed. Furthermore, the calculation method for the shear strength of the structural surface under the action of cyclic shearing is established. The proposed method is further validated through case analysis. The influence of the cutting and filling (produced during the shearing process) on the shear strength of the joint surface cannot be disregarded. The improved model proposed in this study is in good agreement with the experimental results; however, when the improved proposed method is used to estimate the cyclic shear strength of the joint surface where the normal stress is too large, calculation results may contain certain errors