Comparative proteomics reveals essential mechanisms for osmotolerance in Gluconacetobacter diazotrophicus

Plant growth-promoting bacteria are a promising alternative to improve agricultural sustainability. Gluconacetobacter diazotrophicus is an osmotolerant bacterium able to colonize several plant species, including sugarcane, coffee, and rice. Despite its biotechnological potential, the mechanisms controlling such osmotolerance remain unclear. The present study investigated the key mechanisms of resistance to osmotic stress in G. diazotrophicus. The molecular pathways regulated by the stress were investigated by comparative proteomics, and proteins essential for resistance were identified by knock-out mutagenesis. Proteomics analysis led to identify regulatory pathways for osmotic adjustment, de novo saturated fatty acids biosynthesis, and uptake of nutrients. The mutagenesis analysis showed that the lack of AccC protein, an essential component of de novo fatty acid biosynthesis, severely affected G. diazotrophicus resistance to osmotic stress. Additionally, knock-out mutants for nutrients uptake (Δtbdr and ΔoprB) and compatible solutes synthesis (ΔmtlK and ΔotsA) became more sensitive to osmotic stress. Together, our results identified specific genes and mechanisms regulated by osmotic stress in an osmotolerant bacterium, shedding light on the essential role of cell envelope and extracytoplasmic proteins for osmotolerance.     

Increased Nectin-4 levels in chronic ketamine abusers and the relationship with lower urinary tract symptoms

Persistent ketamine use causes susceptibility to addiction and bladder toxicity. We examined the association of lower urinary tract symptoms and levels of Nectin-4, a member of the cell adhesion molecules that is essential for maintaining the urothelium barrier in chronic ketamine abusers. We measured the plasma levels of Nectin-4 in 88 patients with ketamine dependence and 69 controls. Patients with ketamine dependence were assessed for ketamine use variables, psychological symptoms, and lower urinary tract symptoms. We found Nectin-4 levels were increased in ketamine-dependent patients compared to the controls (p < 0.0001). Patients with urinary tract symptoms exhibited lower Nectin-4 levels than those without (p = 0.021). Our results suggest an up-regulation of Nectin-4 following chronic and heavy ketamine use. Patients with ketamine dependence with a compromised upregulation of Nectin-4 are likely to have more severe urinary tract symptoms. The mechanisms underlying the involvement of Nectin-4 in ketamine addiction and bladder toxicity warrant future investigation.     

The association between long-term platelet count and disability-free survival among middle-aged and older Chinese: Evidence from the China Health and Retirement Longitudinal Study

Background and aimsWe aimed to assess the associations of baseline and long-term platelet count (PLT) with disability-free survival (DFS) among middle-aged and older Chinese.Methods and resultsA total of 7296 participants were recruited in the analysis. Updated mean PLT was defined as the mean of the two PLT measurements (4 years between wave 1–3). The long-term status of PLT was defined as persistent low, attenuated, increased and persistent high PLT according to the optimal cut points from the receiver operating characteristic curves of the two PLT measurements, respectively. The primary outcome was DFS, evaluated by the first occurrence of either disability or mortality. During 6-year visit, 1579 participants experienced disability or all-cause mortality. The rates of primary outcome were significantly higher among participants with elevated baseline PLT and updated mean PLT. Multivariable adjusted odds ratios (ORs) and 95% confidence intervals (CIs) of primary outcome were 1.253 (1.049–1.496) for highest baseline PLT tertile and 1.532 (1.124–2.088) for highest updated mean PLT tertile, comparing to the lowest tertiles. Multivariable-adjusted spline regression models showed a linear association of baseline PLT (p_linearity < 0.001) and updated mean PLT (p_linearity = 0.005) with primary outcome. Moreover, participants with persistent high PLT and increased PLT had increased risk of primary outcome (ORs [95% CIs]: 1.825 [1.282–2.597] and 1.767 [1.046–2.985], respectively), compared with the reference of those with persistent low PLT.ConclusionThis study proved elevated baseline PLT, especially long-term persist high or increased PLT was associated with less likelihood of DFS among middle-aged and older Chinese.     

Evolution of gentamicin and arsenite resistance acquisition in Ralstonia pickettii water isolates

Ralstonia pickettii are ubiquitous in water environments. Members of this species are frequently, but not always, resistant to both gentamicin and arsenite. Gentamicin and arsenite co-resistance and the putative molecular mechanisms were investigated. A group of 37 R. pickettii strains isolated from drinking water and hospital wastewater were characterized for gentamicin and arsenite resistance phenotypes, the number and size of plasmids, and screened for genetic elements associated with arsenite tolerance, Integrative and Conjugative Elements (ICEs), among other. The genomes of three representative strains were compared.Most gentamicin resistant (GR) isolates (32/33) were resistant to arsenite, and harbored ICE- and ars operon-related genes. These genetic elements were not detected in any of the five arsenite susceptible strains, regardless of the GR (n = 1) or gentamicin susceptibility (GS) (n = 4) phenotype. The comparison of the genomes of two GR (one resistant and one susceptible to arsenite) and one GS strains suggested that these phenotypes correspond to three phylogroups, distinguished by presence of some genes only in GR isolates, in addition to point mutations in functional genes. The presence of ICEs and ars operon-related genes suggest that arsenite resistance might have been acquired by GR lineages.     

Plasmodium falciparum and Plasmodium vivax specific lactate dehydrogenase: Genetic polymorphism study from Indian isolates

Control and eradication of malaria is hindered by the acquisition of drug resistance by Plasmodium species. This has necessitated a persistent search for novel drugs and more efficient targets. Plasmodium species specific lactate dehydrogenase is one of the potential therapeutic and diagnostic targets, because of its indispensable role in endoerythrocytic stage of the parasite. A target molecule that is highly conserved in the parasite population can be more effectively used in diagnostics and therapeutics, hence, in the present study polymorphism in PfLDH (Plasmodium falciparum specific LDH) and PvLDH (Plasmodium vivax specific LDH) genes was analyzed using PCR-single strand confirmation polymorphism (PCR-SSCP) and sequencing. Forty-six P. falciparum and thirty-five P. vivax samples were screened from different states of India. Our findings have revealed presence of a single PfLDH genotype and six PvLDH genotypes among the studied samples. Interestingly, along with synonymous substitutions, nonsynonymous substitutions were reported to be present for the first time in the PvLDH genotypes. Further, through amino acid sequence alignment and homology modeling studies we observed that the catalytic residues were conserved in all PvLDH genotypes and the nonsynonymous substitutions have not altered the enzyme structure significantly. Evolutionary genetics studies have confirmed that PfLDH and PvLDH loci are under strong purifying selection. Phylogenetic analysis of the pLDH gene sequences revealed that P. falciparum compared to P. vivax, has recent origin. The study therefore supports PfLDH and PvLDH as suitable therapeutic and diagnostic targets as well as phylogenetic markers to understand the genealogy of malaria species.     

基于结构方程模型的男男性行为人群HIV检测行为影响因素研究

目的 分析MSM人群HIV检测行为的影响因素,为制定促进该人群检测HIV的干预措施提供依据。方法 2018年4-6月,通过滚雪球的方法在江苏省无锡市招募MSM研究对象。采取面对面问卷调查的方式,收集研究对象的人口学信息、性行为特征、接受干预服务情况、心理状态和HIV检测情况等。利用结构方程模型对数据进行拟合并对拟合后的模型进行修正,分析各变量间的关系和影响程度。结果 共招募研究对象MSM 410人,认为自己有必要检测HIV占72.9%（299/410）,最近1年HIV检测占69.0%（283/410）。修正后的结构方程模型拟合较好,拟合指数主要结果分别为近似误差均方根（RMSEA）=0.065、规范拟合指数（NFI）=0.946、相对拟合指数（RFI）=0.922、Tucker-Lewis指数（TLI）=0.968、比较拟合指数（CFI）=0.952。模型拟合结果显示,对MSM人群HIV检测行为标准化总效应最高的变量为干预服务,路径系数为0.57;而在干预服务中,HIV检测提醒的因子载荷最大,为0.88。心理状态评分对MSM人群HIV检测行为的标准化总效应呈负向关系,路径系数为-0.33。结论 MSM人群HIV检测意识和检测率有待进一步提高,在采取有效措施促进MSM进行检测的同时,应关注MSM人群心理健康问题。     

Tightly controlled response to oxidative stress; an important factor in the tolerance of Bacteroides fragilis

The exposure of Bacteroides fragilis to highly oxygenated tissues induces an oxidative stress due to a shift from the reduced condition of the gastrointestinal tract to an aerobic environment of host tissues. The potent and effective responses to reactive oxygen species (ROS) make the B. fragilis tolerant to atmospheric oxygen for several days. The response to oxidative stress in B. fragilis is a complicated event that is induced and regulated by different agents. In this review, we will focus on the B. fragilis response to oxidative stress and present an overview of the regulators of responses to oxidative stress in this bacterium.     

Vaccination with recombinant paramyosin in Montanide ISA206 protects against Schistosoma japonicum infection in water buffalo

BackgroundSchistosomiasis japonica is a zoonosis and presents significant public health problems in China and the Philippines. Vaccines targeting domestic animals constitute attractive control measures.MethodsWe conducted three vaccine trials to evaluate the protective efficacy of recombinant full-length paramyosin (rSj97) in water buffalo. Animals were immunized with 3 doses of rSj97 adjuvanted with ISA206 at 250 μg/dose or 500 μg/dose at 4 wk intervals before challenge with 1000 Schistosoma japonicum cercariae. The primary outcome was worm burden assessed by portal perfusion 8–10 weeks post challenge. Safety measures included weight, temperature, body condition score, hemogram and routine assays for hepatic and renal function.ResultsThe three-dose regimen was well tolerated in all three trials. In the first trial, vaccinated buffalo had 51.5% lower worm burden post challenge compared to controls. In the second trial, buffalo immunized with 500 μg/dose of rSj97 had 57.8% lower worm burden compared to controls (p = 0.026). A similar but not significant reduction (60.9%) was observed with animals administered with 250ug rSj97/dose. In the third trial, buffalo immunized with a 500 μg/dose of rSj97 had 57.8% lower worm burden compared to controls (p = 0.014).ConclusionsThese findings indicated that rSj97 is a safe and promising vaccine candidate for schistosomiasis japonica in water buffalo.     

无锡地区H3N2流感病毒流行情况实验室分析

目的 了解无锡地区H3N2流感病毒血凝素基因变异情况和人群中HI抗体水平,分析H3N2流感病毒流行情况.方法 选取2007～2010年哨点医院的H3N2流感病毒进行基因序列比对;选择2008年9月～2009年5月健康体检人群及流感监测哨点医院就诊病例血清140份,2010年9月～2011年1月血清98份进行人群中HI抗体分析.结果 对无锡地区分离的42株H3N2流感病毒的HA基因进行分析,核苷酸序列同源性为98.1％～99.6％,病毒之间的核苷酸序列同源性差异较小.2008年9月～2009年5月人群中H3N2 H1抗体阳性率为86.40％、保护率为84.30％、GMT为1∶58.56,2010年9月～2011年1月人群中为100.00％、100.00％、1∶78.88,两者之间HI抗体阳性率、保护率和GMT差异有统计学意义(P均=0.000).结论 根据对无锡地区2007 ～2011年H3N2亚型流感病毒监测、基因进化分析和人群中抗体保护检测情况分析,推测近期H3N2流感病毒不会有较大流行.     

Combined effects of high sensitivity C-reactive protein and triglyceride–glucose index on risk of cardiovascular disease among middle-aged and older Chinese: Evidence from the China Health and Retirement Longitudinal Study

Background and aimsHigh sensitivity C-reactive protein (hsCRP) and triglyceride glucose (TyG) index were proved to be independent risk factors of cardiovascular disease (CVD). However, individual hsCRP or TyG index might not provide sufficient predictive value on CVD risk. The current study aimed to evaluate the cumulative effect of hsCRP and TyG index on CVD risk prospectively.Methods and resultsA total of 9626 participants were enrolled in the analysis. The TyG index was calculated as ln(triglyceride [mg/dL] × fasting glucose [mg/dL]/2). The primary outcome was new-onset CVD events (cardiac events or stroke), and the secondary outcomes were new-onset cardiac events and stroke, separately. Participants were divided into 4 groups through the median of hsCRP and TyG index. Hazard ratios (HRs) and 95% confidence intervals (CIs) were estimated using multivariable Cox proportion hazard models. From 2013 to 2018, 1730 participants experienced CVD (570 stroke and 1306 cardiac events). Linear associations were found between hsCRP, TyG index, hsCRP/TyG ratio and CVD (all p < 0.05). Compared to participants with low hsCRP/low TyG index, multivariable adjusted HRs (95% CIs) for those with high hsCRP/high TyG index were 1.17 (1.03–1.37) for CVD. No interaction of hsCRP and TyG index was found on CVD (p_-interaction ≥0.05). Furthermore, adding hsCRP and TyG index simultaneously to conventional risk model improved risk reclassification for CVD, stroke and cardiac events (all p < 0.05).ConclusionThe present study suggested combination of hsCRP and TyG index might better improved the ability for risk stratification of CVD among middle-aged and older Chinese.