Correlates of Betel Nut Chewing among Burmese Refugees in Nebraska.

BACKGROUND: Betel nut chewing is an important risk factor for oral cancer, yet there has been little research identifying correlates of betel nut chewing among Burmese refugees in the U.S. METHODS: Based on survey data from 188 Burmese refugees from Nebraska between 2015 and 2016, logistic regression was estimated to identify correlates of betel nut chewing. RESULTS: The prevalence rate of betel nut chewing among participating Burmese refugees in Nebraska was 29%. Relative to Burmese refugees who had an education of less than high school, refugees with higher education were less likely to report betel nut chewing (AOR=0.1, 95% CI (0.02, 0.61)). Refugees who worked full time had higher odds of chewing betel nuts compared to those otherwise (AOR=6.17, 95% CI (1.80, 21.10)). Delaying medication purchase due to cost during the past 12 months was associated with higher odds of betel nut chewing (AOR=5.20, 95% CI (1.02, 26.39)). CONCLUSIONS: Betel nut chewing was common among Burmese refugees in the U.S., yet the odds of betel nut chewing varied across different socioeconomic groups. Health education programs that aim to reduce betel nut chewing might become more cost-effective by disproportionately targeting and serving high-risk groups among Burmese refugees.

SIMONI (Smart Integrated Monitoring) as a novel bioanalytical strategy for water quality assessment: Part II-field feasibility survey.

Because it is impossible to chemically analyze all relevant micropollutants, the implementation of bioanalytical tools is essential to estimate ecological risks of chemical mixtures in regular water-monitoring programs. The first tier of the Smart Integrated Monitoring (SIMONI) strategy, which was described in part I, is based on the combination of passive sampling and bioanalytical measurements. Bioassay responses are compared with effect-based trigger values (EBT), and an overall SIMONI score on all bioassay data was designed to indicate environmental risks. The present study is focused on analyzing the feasibility of the hazard identification tier by evaluating results of 45 field campaigns at sites with different pollution profiles near the city of Amsterdam. A Daphnia assay was performed in situ, while silicon rubber or polar organic chemical integrative sampler (POCIS) extracts were tested with 4 nonspecific (daphnids, algae, bacteria, and cell culture) and 10 specific (9 Chemical Activated Luciferase Gene Expression [CALUX] assays and antibiotics scan) bioassays. Sensitivity analyses demonstrated the relevance of 2 classification variables in the SIMONI score formula on all bioanalytical data. The model indicated increased risks for the ecosystem at surface waters in greenhouse areas and undiluted wastewater-treatment plant (WWTP) effluents. The choice of testing specific bioassays on either polar or nonpolar passive sampling extracts is cost-effective and still provided meaningful insights on micropollutant risks. Statistical analyses revealed that the model provides a relevant overall impact assessment based on bioassay responses. Data analyses on the chemically determined mixture toxic pressure and bioanalytical methods provided similar insights in relative risk ranking of water bodies. The SIMONI combination of passive sampling and bioanalytical testing appears to be a feasible strategy to identify chemical hazards. Environ Toxicol Chem 2017;36:2400-2416. © 2017 SETAC.

SIMONI (smart integrated monitoring) as a novel bioanalytical strategy for water quality assessment: Part i-model design and effect-based trigger values.

It is virtually impossible to reliably assess water quality with target chemical analyses only. Therefore, a complementary effect-based risk assessment by bioanalyses on mixtures of bioavailable micropollutants is proposed: the Smart Integrated Monitoring (SIMONI) strategy. The goal of this strategy is to obtain more reliable information on the water quality to select optimum measures for improvement. The SIMONI strategy is 2-tiered. Tier 1 is a bioanalytical hazard identification of sites. A tier 2 ecological risk assessment is carried out only at a limited number of sites where increased hazards are detected in tier 1. Tier 2 will be customized, based on tier 1 evaluation and additional knowledge of the aquatic system. The present study focuses on the tier 1 bioanalytical hazard identification to distinguish "hot spots" of chemical pollution. First, a selection was made of relevant and cost-effective bioanalytical endpoints to cover a wide spectrum of micropollutant modes of action. Specific endpoints may indicate which classes of chemicals might cause adverse effects. Second, effect-based trigger values (EBT) were derived for these bioassays to indicate potential ecological risks. Comparison of EBT with bioassay responses should discriminate sites exhibiting different chemical hazards. Third, a model was designed to estimate the overall risks for aquatic ecosystems. The associated follow-up for risk management is a "toxicity traffic light" system: green, low hazard (no action required); orange, potential risk (further research needed); and red, high risk (mitigation measures). Thanks to cost-effectiveness, flexibility, and relevance, the SIMONI strategy has the potential to become the first bioanalytical tool to be applied in regular water quality monitoring programs. Environ Toxicol Chem 2017;36:2385-2399. © 2017 SETAC.

Bioassay battery interlaboratory investigation of emerging contaminants in spiked water extracts - Towards the implementation of bioanalytical monitoring tools in water quality assessment and monitoring.

Bioassays are particularly useful tools to link the chemical and ecological assessments in water quality monitoring. Different methods cover a broad range of toxicity mechanisms in diverse organisms, and account for risks posed by non-target compounds and mixtures. Many tests are already applied in chemical and waste assessments, and stakeholders from the science-police interface have recommended their integration in regulatory water quality monitoring. Still, there is a need to address bioassay suitability to evaluate water samples containing emerging pollutants, which are a current priority in water quality monitoring. The presented interlaboratory study (ILS) verified whether a battery of miniaturized bioassays, conducted in 11 different laboratories following their own protocols, would produce comparable results when applied to evaluate blinded samples consisting of a pristine water extract spiked with four emerging pollutants as single chemicals or mixtures, i.e. triclosan, acridine, 17α-ethinylestradiol (EE2) and 3-nitrobenzanthrone (3-NBA). Assays evaluated effects on aquatic organisms from three different trophic levels (algae, daphnids, zebrafish embryos) and mechanism-specific effects using in vitro estrogenicity (ER-Luc, YES) and mutagenicity (Ames fluctuation) assays. The test battery presented complementary sensitivity and specificity to evaluate the different blinded water extract spikes. Aquatic organisms differed in terms of sensitivity to triclosan (algae > daphnids > fish) and acridine (fish > daphnids > algae) spikes, confirming the complementary role of the three taxa for water quality assessment. Estrogenicity and mutagenicity assays identified with high precision the respective mechanism-specific effects of spikes even when non-specific toxicity occurred in mixture. For estrogenicity, although differences were observed between assays and models, EE2 spike relative induction EC50 values were comparable to the literature, and E2/EE2 equivalency factors reliably reflected the sample content. In the Ames, strong revertant induction occurred following 3-NBA spike incubation with the TA98 strain, which was of lower magnitude after metabolic transformation and when compared to TA100. Differences in experimental protocols, model organisms, and data analysis can be sources of variation, indicating that respective harmonized standard procedures should be followed when implementing bioassays in water monitoring. Together with other ongoing activities for the validation of a basic bioassay battery, the present study is an important step towards the implementation of bioanalytical monitoring tools in water quality assessment and monitoring.

Development of flavonoid-based inverse agonists of the key signaling receptor US28 of human cytomegalovirus.

A series of 31 chalcone- and flavonoid-based derivatives were synthesized in good overall yields and screened for their inverse agonist activity on the US28 receptor of human cytomegalovirus (HCMV). With one exception (e.g., 2-(5-bromo-2-methoxyphenyl)-3-hydroxy-4H-chromen-4-one), halogen-substituted flavonoids were typically more potent inverse agonists than their related hydro derivatives. While toxicity could be used to partially explain the inverse agonist activity of some members of the series, 5-(benzyloxy)-2-(5-bromo-2-methoxyphenyl)-4H-chromen-4-one (11b) acted on the US28 receptor as a nontoxic, inverse agonist. The full inverse agonism (efficacy, -89%) and potency (EC50 = 3.5 µM) observed with flavonoid 11b is especially important as it provides both a new tool to study US28 signaling and a potential platform for the future development of HCMV-targeting drugs.

Effects of salt, polyethylene glycol, and locked nucleic acids on the thermodynamic stabilities of consecutive terminal adenosine mismatches in RNA duplexes.

Consecutive terminal mismatches add thermodynamic stability to RNA duplexes and occur frequently in microRNA-mRNA interactions. Accurate thermodynamic stabilities of consecutive terminal mismatches contribute to the development of specific, high-affinity siRNA therapeutics. Consecutive terminal adenosine mismatches (TAMS) are studied at different salt concentrations, with polyethylene glycol cosolutes, and with locked nucleic acid (LNA) substitutions. These measurements provide benchmarks for the application of thermodynamic predictions to different physiological or therapeutic conditions. The salt dependence for RNA duplex stability is similar for TAMS, internal loops, and Watson-Crick duplexes. A unified model for predicting the free energy of an RNA duplex with or without loops and mismatches at lower sodium concentrations is presented. The destabilizing effects of PEG 200 are larger for TAMS than internal loops or Watson-Crick duplexes, which may result from different base stacking conformations, dynamics, and water hydration. In contrast, LNA substitutions stabilize internal loops much more than TAMS. Surprisingly, the average per adenosine increase in stability for LNA substitutions in internal loops is -1.82 kcal/mol and only -0.20 kcal/mol for TAMS. The stabilities of TAMS and internal loops with LNA substitutions have similar favorable free energies. Thus, the unfavorable free energy of adenosine internal loops is largely an entropic effect. The favorable stabilities of TAMS result mainly from base stacking. The ability of RNA duplexes to form extended terminal mismatches in the absence of proteins such as argonaute and identifying the enthalpic contributions to terminal mismatch stabilities provide insight into the physical basis of microRNA-mRNA molecular recognition and specificity.

Validation of the REA bioassay to detect estrogenic activity in the water cycle.

Endocrine disrupting compounds (EDCs) with estrogenic potency contaminate water and might eventually cause adverse effects to the aquatic environment. Many estrogenic compounds are not completely removed by wastewater treatment systems and, together with the run-off from agricultural areas, they enter surface waters. Chemical analytical methods to determine these compounds are usually expensive and laborious. Therefore, screening bioassays which are able to detect compounds based on their effects offer a solution for prior selection of samples that need to be chemically analyzed. In this study, the REA (RIKILT yeast Estrogen bioAssay), which has been developed to detect estrogenic compounds in calf urine and animal feed at RIKILT, is validated at the Water Board Laboratory of Waterproef for water samples. According to EC Decision 2002/657, detection capability CCß, specificity and stability have to be determined for the internal validation of a qualitative screening test. In addition, surface water and effluent samples were analyzed to further demonstrate the applicability of the validated test procedure. Results demonstrate that the REA assay is reproducible and specific for estrogenic compounds in water and meets the criteria as prescribed in EC Decision 2002/657. The assay was sensitive enough to detect estrogenic activity of pollutants in water with a limit of quantification (LOQ) below 1 ng EEQ/L. This means that samples can be compared with preliminary threshold levels for drinking water and surface waters (7 and 1 ng EEQ/L, respectively). The stability of estrogenic activity in water samples is at least 4 weeks, when stored at 4 °C.

Consecutive terminal GU pairs stabilize RNA helices.

Consecutive GU pairs at the ends of RNA helices provide significant thermodynamic stability between -1.0 and -3.8 kcal/mol at 37 °C, which is equivalent to approximately 2 orders of magnitude in the value of a binding constant. The thermodynamic stabilities of GU pairs depend on the sequence, stacking orientation, and position in the helix. In contrast to GU pairs in the middle of a helix that may be destabilizing, all consecutive terminal GU pairs contribute favorable thermodynamic stability. This work presents measured thermodynamic stabilities for 30 duplexes containing two, three, or four consecutive GU pairs at the ends of RNA helices and a model to predict the thermodynamic stabilities of terminal GU pairs. Imino proton NMR spectra show that the terminal GU nucleotides form hydrogen-bonded pairs. Different orientations of terminal GU pairs can have different conformations with equivalent thermodynamic stabilities. These new data and prediction model will help improve RNA secondary structure prediction, identification of miRNA target sequences with GU pairs, and efforts to understand the fundamental physical forces directing RNA structure and energetics.