Crystal structures of a halophilic archaeal malate synthase from Haloferax volcanii and comparisons with isoforms A and G

<p>Abstract</p> <p>Background</p> <p>Malate synthase, one of the two enzymes unique to the glyoxylate cycle, is found in all three domains of life, and is crucial to the utilization of two-carbon compounds for net biosynthetic pathways such as gluconeogenesis. In addition to the main isoforms A and G, so named because of their differential expression in <it>E. coli </it>grown on either acetate or glycolate respectively, a third distinct isoform has been identified. These three isoforms differ considerably in size and sequence conservation. The A isoform (MSA) comprises ~530 residues, the G isoform (MSG) is ~730 residues, and this third isoform (MSH-halophilic) is ~430 residues in length. Both isoforms A and G have been structurally characterized in detail, but no structures have been reported for the H isoform which has been found thus far only in members of the halophilic Archaea.</p> <p>Results</p> <p>We have solved the structure of a malate synthase H (MSH) isoform member from <it>Haloferax volcanii </it>in complex with glyoxylate at 2.51 Å resolution, and also as a ternary complex with acetyl-coenzyme A and pyruvate at 1.95 Å. Like the A and G isoforms, MSH is based on a β8/α8 (TIM) barrel. Unlike previously solved malate synthase structures which are all monomeric, this enzyme is found in the native state as a trimer/hexamer equilibrium. Compared to isoforms A and G, MSH displays deletion of an N-terminal domain and a smaller deletion at the C-terminus. The MSH active site is closely superimposable with those of MSA and MSG, with the ternary complex indicating a nucleophilic attack on pyruvate by the enolate intermediate of acetyl-coenzyme A.</p> <p>Conclusions</p> <p>The reported structures of MSH from <it>Haloferax volcanii </it>allow a detailed analysis and comparison with previously solved structures of isoforms A and G. These structural comparisons provide insight into evolutionary relationships among these isoforms, and also indicate that despite the size and sequence variation, and the truncated C-terminal domain of the H isoform, the catalytic mechanism is conserved. Sequence analysis in light of the structure indicates that additional members of isoform H likely exist in the databases but have been misannotated.</p

Climate Change and Utah Ski Resorts: Impacts, Perceptions, and Adaptation Strategies

Climate change is a threat to ski resorts, the ski industry, and mountain communities that rely on ski tourism. Ski resorts may be able to mitigate some of the social and economic impacts caused by climate change with proactive adaptation strategies. Using historical weather data, future climate projections, and interviews with ski resort managers in Utah (United States), this research investigates the effects of climate change on ski resorts across the state. We examine temperature change at all resorts within the state from 1980–2018 and climate projections from 2021–2100 under different climate change scenarios (RCPs 2.6, 4.5, and 8.5). We also report on semistructured interviews with resort managers to provide insights into how resort leadership perceives the impacts of climate change, is implementing adaptation strategies, and is addressing barriers to adaptation. Many resorts in Utah are warming faster than global averages, and minimum temperatures are rising faster than maximum temperatures. By the end of the century, winter (December–March) minimum daily temperatures in Utah could warm an additional 6.0°C under the RCP 8.5 scenario near northern Utah resorts and 6.6°C near southern Utah resorts. Resort managers are concerned about shorter season lengths, shifting ski seasons, less snow cover, and poorer snow quality. Many resorts are already adapting, with the most common adaptations being snowmaking and diversifying outdoor recreation offerings (particularly during the summer and shoulder seasons). Barriers to adaptation reported by managers include financial costs, adequate water availability for snowmaking, and uncertainty about climate change projections. Climate change is already impacting Utah ski resorts, but adaptation practices can reduce the negative impacts to some degree at most resorts

15th Annual Environmental Law Institute

Materials from the 15th Annual Environmental Law Institute held by UK/CLE in March 1999

Proteomic Analysis of Resistance of Gram-Negative Bacteria to Chlorhexidine and Impacts on Susceptibility to Colistin, Antimicrobial Peptides, and Ceragenins

Use of chlorhexidine in clinical settings has led to concerns that repeated exposure of bacteria to sub-lethal doses of chlorhexidine might result in chlorhexidine resistance and cross resistance with other cationic antimicrobials including colistin, endogenous antimicrobial peptides (AMPs) and their mimics, ceragenins. We have previously shown that colistin-resistant Gram-negative bacteria remain susceptible to AMPs and ceragenins. Here, we investigated the potential for cross resistance between chlorhexidine, colistin, AMPs and ceragenins by serial exposure of standard strains of Gram-negative bacteria to chlorhexidine to generate resistant populations of organisms. Furthermore, we performed a proteomics study on the chlorhexidine-resistant strains and compared them to the wild-type strains to find the pathways by which bacteria develop resistance to chlorhexidine. Serial exposure of Gram-negative bacteria to chlorhexidine resulted in four- to eight-fold increases in minimum inhibitory concentrations (MICs). Chlorhexidine-resistant organisms showed decreased susceptibility to colistin (8- to 32-fold increases in MICs) despite not being exposed to colistin. In contrast, chlorhexidine-resistant organisms had the same MICs as the original strains when tested with representative AMPs (LL-37 and magainin I) and ceragenins (CSA-44 and CSA-131). These results imply that there may be a connection between the emergence of highly colistin-resistant Gram-negative pathogens and the prevalence of chlorhexidine usage. Yet, use of chlorhexidine may not impact innate immune defenses (e.g., AMPs) and their mimics (e.g., ceragenins). Here, we also show that chlorhexidine resistance is associated with upregulation of proteins involved in the assembly of LPS for outer membrane biogenesis and virulence factors in Pseudomonas aeruginosa. Additionally, resistance to chlorhexidine resulted in elevated expression levels of proteins associated with chaperones, efflux pumps, flagella and cell metabolism. This study provides a comprehensive overview of the evolutionary proteomic changes in P. aeruginosa following exposure to chlorhexidine and colistin. These results have important clinical implications considering the continuous application of chlorhexidine in hospitals that could influence the emergence of colistin-resistant strains

Reliability and validity of the Adolescent Stress Questionnaire in a sample of European adolescents--the HELENA study

BACKGROUND: Since stress is hypothesized to play a role in the etiology of obesity during adolescence, research on associations between adolescent stress and obesity-related parameters and behaviours is essential. Due to lack of a well-established recent stress checklist for use in European adolescents, the study investigated the reliability and validity of the Adolescent Stress Questionnaire (ASQ) for assessing perceived stress in European adolescents. METHODS: The ASQ was translated into the languages of the participating cities (Ghent, Stockholm, Vienna, Zaragoza, Pecs and Athens) and was implemented within the HELENA cross-sectional study. A total of 1140 European adolescents provided a valid ASQ, comprising 10 component scales, used for internal reliability (Cronbach α) and construct validity (confirmatory factor analysis or CFA). Contributions of socio-demographic (gender, age, pubertal stage, socio-economic status) characteristics to the ASQ score variances were investigated. Two-hundred adolescents also provided valid saliva samples for cortisol analysis to compare with the ASQ scores (criterion validity). Test-retest reliability was investigated using two ASQ assessments from 37 adolescents. RESULTS: Cronbach α-values of the ASQ scales (0.57 to 0.88) demonstrated a moderate internal reliability of the ASQ, and intraclass correlation coefficients (0.45 to 0.84) established an insufficient test-retest reliability of the ASQ. The adolescents' gender (girls had higher stress scores than boys) and pubertal stage (those in a post-pubertal development had higher stress scores than others) significantly contributed to the variance in ASQ scores, while their age and socio-economic status did not. CFA results showed that the original scale construct fitted moderately with the data in our European adolescent population. Only in boys, four out of 10 ASQ scale scores were a significant positive predictor for baseline wake-up salivary cortisol, suggesting a rather poor criterion validity of the ASQ, especially in girls. CONCLUSIONS: In our European adolescent sample, the ASQ had an acceptable internal reliability and construct validity and the adolescents' gender and pubertal stage systematically contributed to the ASQ variance, but its test-retest reliability and criterion validity were rather poor. Overall, the utility of the ASQ for assessing perceived stress in adolescents across Europe is uncertain and some aspects require further examination.The HELENA Study takes place with the financial support of the European Community Sixth RTD Framework Programme (Contract FOOD-CT-2005-007034)

"Sleep disparity" in the population: poor sleep quality is strongly associated with poverty and ethnicity

<p>Abstract</p> <p>Background</p> <p>Little is known about the social determinants of sleep attainment. This study examines the relationship of race/ethnicity, socio-economic status (SES) and other factors upon sleep quality.</p> <p>Methods</p> <p>A cross-sectional survey of 9,714 randomly selected subjects was used to explore sleep quality obtained by self-report, in relation to socioeconomic factors including poverty, employment status, and education level. The primary outcome was poor sleep quality. Data were collected by the Philadelphia Health Management Corporation.</p> <p>Results</p> <p>Significant differences were observed in the outcome for race/ethnicity (African-American and Latino versus White: unadjusted OR = 1.59, 95% CI 1.24-2.05 and OR = 1.65, 95% CI 1.37-1.98, respectively) and income (below poverty threshold, unadjusted OR = 2.84, 95%CI 2.41-3.35). In multivariable modeling, health indicators significantly influenced sleep quality most prominently in poor individuals. After adjusting for socioeconomic factors (education, employment) and health indicators, the association of income and poor sleep quality diminished, but still persisted in poor Whites while it was no longer significant in poor African-Americans (adjusted OR = 1.95, 95% CI 1.47-2.58 versus OR = 1.16, 95% CI 0.87-1.54, respectively). Post-college education (adjusted OR = 0.47, 95% CI 0.31-0.71) protected against poor sleep.</p> <p>Conclusions</p> <p>A "sleep disparity" exists in the study population: poor sleep quality is strongly associated with poverty and race. Factors such as employment, education and health status, amongst others, significantly mediated this effect only in poor subjects, suggesting a differential vulnerability to these factors in poor relative to non-poor individuals in the context of sleep quality. Consideration of this could help optimize targeted interventions in certain groups and subsequently reduce the adverse societal effects of poor sleep.</p

Gene Expression Signature Analysis Identifies Vorinostat as a Candidate Therapy for Gastric Cancer

Gastric cancer continues to be one of the deadliest cancers in the world and therefore identification of new drugs targeting this type of cancer is thus of significant importance. The purpose of this study was to identify and validate a therapeutic agent which might improve the outcomes for gastric cancer patients in the future. manifested a reversed pattern.We showed that analysis of gene expression signature may represent an emerging approach to discover therapeutic agents for gastric cancer, such as vorinostat. The observation of altered gene expression after vorinostat treatment may provide the clue to identify the molecular mechanism of vorinostat and those patients likely to benefit from vorinostat treatment