The two-step strategy for enhancing the specific activity and thermostability of alginate lyase AlyG2 with mechanism for improved thermostability.

To overcome the trade-off challenge encountered in the engineering of alginate lyase AlyG2 from Seonamhaeicola algicola Gy8T and to expand its potential industrial applications, we devised a two-step strategy encompassing activity enhancement followed by thermal stability engineering. To enhance the specific activity of efficient AlyG2, we strategically substituted residues with bulky steric hindrance proximal to the active pocket with glycine or alanine. This led to the generation of three promising positive mutants, with particular emphasis on the T91S mutant, exhibiting a 1.91-fold specific activity compared to the wild type. To mitigate the poor thermal stability of T91S, mutants with negative ΔΔG values in the thermal flexibility region were screened out. Notably, the S72Ya mutant not only displayed 17.96 % further increase in specific activity but also exhibited improved stability compared to T91S, manifesting as a remarkable 30.97 % increase in relative activity following a 1-hour incubation at 42 °C. Furthermore, enhanced kinetic stability was observed. To gain deeper insights into the mechanism underlying the enhanced thermostability of the S72Ya mutant, we conducted molecular dynamics simulations, principal component analysis (PCA), dynamic cross-correlation map (DCCM), and free energy landscape (FEL) analysis. The results unveiled a reduction in the flexibility of the surface loop, a stronger correlation dynamic and a narrower motion subspace in S72Ya system, along with the formation of more stable hydrogen bonds. Collectively, our findings suggest amino acids substitutions resulting in smaller side chains proximate to the active site can positively impact enzyme activity, while reducing the flexibility of surface loops emerges as a pivotal factor in conferring thermal stability. These insights offer valuable guidance and a framework for the engineering of other enzyme types.

Silver Nanoparticles Encapped by Dihydromyricetin: Optimization of Green Synthesis, Characterization, Toxicity, and Anti-MRSA Infection Activities for Zebrafish (Danio rerio).

To achieve the environmentally friendly and rapid green synthesis of efficient and stable AgNPs for drug-resistant bacterial infection, this study optimized the green synthesis process of silver nanoparticles (AgNPs) using Dihydromyricetin (DMY). Then, we assessed the impact of AgNPs on zebrafish embryo development, as well as their therapeutic efficacy on zebrafish infected with Methicillin-resistant Staphylococcus aureus (MRSA). Transmission electron microscopy (TEM) and dynamic light-scattering (DLS) analyses revealed that AgNPs possessed an average size of 23.6 nm, a polymer dispersity index (PDI) of 0.197 ± 0.0196, and a zeta potential of -18.1 ± 1.18 mV. Compared to other published green synthesis products, the optimized DMY-AgNPs exhibited smaller sizes, narrower size distributions, and enhanced stability. Furthermore, the minimum concentration of DMY-AgNPs required to affect zebrafish hatching and survival was determined to be 25.0 µg/mL, indicating the low toxicity of DMY-AgNPs. Following a 5-day feeding regimen with DMY-AgNP-containing food, significant improvements were observed in the recovery of the gills, intestines, and livers in MRSA-infected zebrafish. These results suggested that optimized DMY-AgNPs hold promise for application in aquacultures and offer potential for further clinical use against drug-resistant bacteria.

Do Real-Time Strategy Video Gamers Have Better Attentional Control?

OBJECTIVE: Do real-time strategy (RTS) video gamers have better attentional control? To examine this issue, we tested experienced versus inexperienced RTS video gamers on multi-object tracking tasks (MOT) and dual-MOT tasks with visual or auditory secondary tasks (dMOT). We employed a street-crossing task with a visual working memory task as a secondary task in a virtual reality (VR) environment to examine any generalized attentional advantage. BACKGROUND: Similar to action video games, RTS video games require players to switch attention between multiple visual objects and views. However, whether the attentional control advantage is limited by sensory modalities or generalizes to real-life tasks remains unclear. METHOD: In study 1, 25 RTS video game players (SVGP) and 25 non-video game players (NVGP) completed the MOT task and two dMOT tasks. In study 2, a different sample with 25 SVGP and 25 NVGP completed a simulated street-crossing task with the visual dual task in a VR environment. RESULTS: After controlling the effects of the speed-accuracy trade-off, SVGP showed better performance than NVGP in the MOT task and the visual dMOT task, but SVGP did not perform better in either the auditory dMOT task or the street-crossing task. CONCLUSION: RTS video gamers had better attentional control in visual computer tasks, but not in the auditory tasks and the VR tasks. Attentional control benefits associated with RTS video game experience may be limited by sensory modalities, and may not translate to performance benefits in real-life tasks.

A review of psychological interventions for internet addiction.

Internet addiction (IA) may constitute a widespread and serious mental problem. Previous reviews have not fully considered potential factors that may contribute to therapeutic outcomes or predict behavioral changes. Such information is relevant to understand the active ingredients of interventions and to develop more efficacious treatments that target features of IA. This systematic review was designed to relate theories of IA to treatments, describe studies of psychotherapies for IA, and propose a model of addiction and interventions based on extant studies. A computer database search of PubMed, PsychINFO, ScienceDirect, China National Knowledge Infrastructure, and Google Scholar was conducted to identify all available research evidence on psychological treatments for IA (N = 31 studies). Among these psychological interventions, the targeted reduction of addiction-related impulsivity and craving, improvement of cognitive maladjustment, and alleviation of family problems have been investigated in IA interventions. The targeted domains and intervention methods are not mutually exclusive, and further research is needed to demonstrate the effective components and mechanisms of action for treatments of IA. Such research will help generate more efficacious evidence-based interventions.

Terminal uridyltransferase 7 regulates TLR4-triggered inflammation by controlling Regnase-1 mRNA uridylation and degradation.

Different levels of regulatory mechanisms, including posttranscriptional regulation, are needed to elaborately regulate inflammatory responses to prevent harmful effects. Terminal uridyltransferase 7 (TUT7) controls RNA stability by adding uridines to its 3' ends, but its function in innate immune response remains obscure. Here we reveal that TLR4 activation induces TUT7, which in turn selectively regulates the production of a subset of cytokines, including Interleukin 6 (IL-6). TUT7 regulates IL-6 expression by controlling ribonuclease Regnase-1 mRNA (encoded by Zc3h12a gene) stability. Mechanistically, TLR4 activation causes TUT7 to bind directly to the stem-loop structure on Zc3h12a 3'-UTR, thereby promotes Zc3h12a uridylation and degradation. Zc3h12a from LPS-treated TUT7-sufficient macrophages possesses increased oligo-uridylated ends with shorter poly(A) tails, whereas oligo-uridylated Zc3h12a is significantly reduced in Tut7-/- cells after TLR4 activation. Together, our findings reveal the functional role of TUT7 in sculpting TLR4-driven responses by modulating mRNA stability of a selected set of inflammatory mediators.

Blood lead levels of children in urban and suburban areas in China (1997-2015): Temporal and spatial variations and influencing factors.

Blood lead (Pb) poisoning is a worldwide heath problem, especially in developing countries. As the largest developing country in the world, China faces severe health challenges, in particular the threat of blood Pb poisoning. In this study, the temporal trend of Chinese children's blood lead levels (BLLs) and blood lead poisoning incidence (BLPI) (percentage of BLL>100µg/L) and its influencing factors were investigated. We collected articles on children's BLLs published from 1997 to 2017 with sampling time from 1997 to 2015 by searching the databases of VIP Medical Information System (VMIS), China National Knowledge Infrastructure (CNKI) and Wanfang Data. After a rigorous investigation, 259 articles with eligible inclusion criteria were reviewed. Meanwhile, the data of Pb concentrations in the soil of 23 cities and the annual mean PM10 (particulate matter<10µm) concentrations of 24 provincial cities were collected. The temporal trend of children's BLLs and BLPIs could be divided into three stages: upward trend from 1997 to 2000, downward trend from 2001 to 2013, and upward trend from 2014 to 2015. The decline of BLLs from 2001 was primarily due to the phasing out of leaded gasoline since 2000 in China, while the descending air quality could explain the upward trend of BLLs in the period from 2014 to 2015. The correlation and regression analysis indicated that soil and air were two major pathways of Pb exposure for children in China. Although a noticeable decrease has been shown, the Chinese children's BLLs were still significantly higher than the levels of developed countries. We highly recommended that the critical value of blood Pb poisoning should be lowered to 50µg/L in China. Guidelines on the prevention and management of blood Pb poisoning are needed in China.

The roles of PAI-1 gene polymorphisms in atherosclerotic diseases: A systematic review and meta-analysis involving 149,908 subjects.

BACKGROUND: The roles of plasminogen activator inhibitor-1 (PAI-1) gene polymorphisms in atherosclerotic diseases were intensively analyzed, but the results of these studies were inconsistent. Therefore, we performed this study to better assess the relationship between PAI-1 genetic variations and atherosclerosis. METHODS: Eligible studies were searched in PubMed, Medline, Embase and Web of Science. Odds ratios (ORs) with 95% confidence intervals (CIs) were used to assess relationship between PAI-1 polymorphisms and atherosclerotic diseases. RESULTS: Ninety-nine studies involving 62,739 cases and 87,169 controls were finally included. Significant associations with the risk of atherosclerosis were detected for the rs2227631 polymorphism in the dominant model (95% CI 0.84-1.00), for the rs1799889 polymorphism in the dominant (95% CI 1.01-1.18), recessive (95% CI 0.90-0.98) and allele (95% CI 1.01-1.12) models. Further subgroup analyses based on type of disease and ethnicity of participants suggested that the rs2227631 polymorphism was significantly associated with the risk of coronary artery disease in the dominant (95% CI 0.71-0.94) and allele (95% CI 0.80-0.94) models, whereas the rs1799889 polymorphism was significantly associated with the risk of myocardial infarction (dominant model: 95% CI 1.09-1.57; recessive model: 95% CI 0.71-0.96; allele model: 95% CI 1.05-1.28) and cerebral infarction (dominant model: 95% CI 1.68-3.51; additive model: 95% CI 0.39-0.77; allele model: 95% CI 1.23-2.00). Moreover, the rs1799889 polymorphism was also significantly correlated with the risk of atherosclerosis in both Asians (dominant model: 95% CI 1.10-1.83; allele model: 95% CI 1.03-1.41) and Caucasians (recessive model: 95% CI 0.87-0.97; allele model: 95% CI 1.01-1.12). CONCLUSION: In conclusion, our findings indicate that PAI-1 rs2227631 and rs1799889 polymorphisms may serve as genetic biomarkers of atherosclerotic diseases.

Genetic analysis of colon tumors induced by a dietary carcinogen PhIP in CYP1A humanized mice: Identification of mutation of ß-catenin/Ctnnb1 as the driver gene for the carcinogenesis.

Replacing mouse Cyp1a with human CYP1A enables the humanized CYP1A mice to mimic human metabolism of the dietary carcinogen, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), by N(2) -hydroxylation to a proximate carcinogen. Our previous study demonstrated that PhIP, combined with the dextrin sulfate sodium (DSS)-induced colitis, induces colon carcinogenesis in hCYP1A mice. Here, we employed whole exome sequencing and found multiple gene mutations in PhIP/DSS-induced colon tumors. Mutations in the exon 3 of Ctnnb1/ß-catenin, however, were the predominant events. We further sequenced the key fragments of Apc, Ctnnb1, and Kras, because mutations of these genes in the humans are commonly found as the drivers of colorectal cancer. Mutations on either codon 32 or 34 in the exon 3 of Ctnnb1 were found in 39 out of 42 tumors, but no mutation was found in either Apc or Kras. The sequence context of codons 32 and 34 suggests that PhIP targets +3G in a TGGA motif of Ctnnb1. Since mutations that activate Wnt signal is a major driving force for human colorectal cancers, we conclude that the mutated ß-catenin is the driver in PhIP/DSS-induced colon carcinogenesis. This result suggests that the colon tumors in hCYP1A mice mimic human colorectal carcinogenesis not only in the dietary etiology involving PhIP, but also in the aberrant activation of the Wnt signaling pathway as the driving force.