Differential DNA accessibility to polymerase enables 30-minute phenotypic β-lactam antibiotic susceptibility testing of carbapenem-resistant Enterobacteriaceae

The rise in carbapenem-resistant Enterobacteriaceae (CRE) infections has created a global health emergency, underlining the critical need to develop faster diagnostics to treat swiftly and correctly. Although rapid pathogen-identification (ID) tests are being developed, gold-standard antibiotic susceptibility testing (AST) remains unacceptably slow (1–2 d), and innovative approaches for rapid phenotypic ASTs for CREs are urgently needed. Motivated by this need, in this manuscript we tested the hypothesis that upon treatment with β-lactam antibiotics, susceptible Enterobacteriaceae isolates would become sufficiently permeabilized, making some of their DNA accessible to added polymerase and primers. Further, we hypothesized that this accessible DNA would be detectable directly by isothermal amplification methods that do not fully lyse bacterial cells. We build on these results to develop the polymerase-accessibility AST (pol-aAST), a new phenotypic approach for β-lactams, the major antibiotic class for gram-negative infections. We test isolates of the 3 causative pathogens of CRE infections using ceftriaxone (CRO), ertapenem (ETP), and meropenem (MEM) and demonstrate agreement with gold-standard AST. Importantly, pol-aAST correctly categorized resistant isolates that are undetectable by current genotypic methods (negative for β-lactamase genes or lacking predictive genotypes). We also test contrived and clinical urine samples. We show that the pol-aAST can be performed in 30 min sample-to-answer using contrived urine samples and has the potential to be performed directly on clinical urine specimens

HEALTHY BITES: A CHALLENGE-BASED WORKSITE WELLNESS NUTRITION PROGRAM

Purpose: The worksite health promotion literature has shown that successful nutrition communication programs call for making small changes to diet, one at a time. The use of observational learning (role models) and a “challenge” scenario for behavior change is supported by Social Cognitive Theory. The significance of these small changes in worksites is underscored by a recent Harvard Business Review article suggesting workplace wellness programs can return up $2.71 on every$1.00 invested. The purpose of the proposed program, Healthy Bites, is to increase knowledge of employees regarding the impact of specific nutrition behaviors related to reducing cancer risk and improving weight status. The relationships between types of activities attended and/or tools used with degree of behavior change will also be evaluated. Methods: Eligible participants are MD Anderson Cancer Center employees. Participants will be asked to complete monthly nutrition challenges during 2013 to possibly reduce cancer risk and improve weight status. Challenge topics include: eating breakfast, not skipping meals, consuming whole grains, following the MyPlate diagram, eating less red meat and processed meat, eating more plant-based protein, drinking more water, eating out less, increasing fruit and vegetables, limiting sodium, and drinking less alcohol. Each month participants will receive printed tools, tips and recipes via email. Participants will also be able to attend cooking demonstrations and informational lectures, as well as read stories about fellow employees who have successfully completed the challenges. Results: (Evaluation): Surveys will be distributed to participants at program initiation (January 2013), the mid-point (July 2013) and at conclusion (December 2013). The surveys will evaluate/assess: nutrition knowledge, nutrition habits, enhancements to improve the challenges, likeability of program, number of challenges completed, whether families participating together were more successful, and usefulness of Healthy Bites tools and content. Conclusions: Healthy Bites will evaluate how a nutrition behavior change program can be successfully implemented in a large hospital-based workplace setting to improve nutrition knowledge and habits among employees

Using GIS-Based, Regional Extent Habitat Suitability Modeling to Identify Conservation Priority Areas: A Case Study of the Louisiana Black Bear in East Texas

State and federal recovery plans mandate that priority areas for future population expansion be identified within the historical range of the Louisiana black bear (Ursus americanus luteolus). Despite the presence of potentially suitable habitat in east Texas and expanding populations in adjacent states, quantitative estimates of regional habitat suitability do not exist. We developed a regional extent habitat suitability index (HSI) model in a geographic information system (GIS) to evaluate year-round habitat requirements for black bears in the 43,530-km2 south black bear recovery zone in southeastern Texas. We measured hard and soft mast production, understory vegetation density, and tree den availability at 516 survey points in 38 habitat classes (82% of the total area in the south recovery zone). We developed geospatial models for summer food availability; fall food availability, diversity, and productivity; protection cover, tree den availability, distance to roads, and human development zones and calculated HSI scores per pixel in a continuous dataset. Habitat suitability scores ranged from 0.00 to 0.76 throughout southeastern Texas. Highly (20,700 ha (mean HSI=0.5) capable of sustaining viable black bear populations. These units ranged from 62,844 ha to 124,808 ha in size and suitable habitat pixels within units ranged from 0.58 to 0.60 in mean HSI scores. Recovery unit scores were comparable to those previously reported for occupied bear range in the southeastern United States and acreages of suitable habitat exceeded those estimated to support existing Louisiana black bear populations

Prodigious polyphyly in Pleuroceridae (Gastropoda: Cerithioidea)

Phylogenomic studies with hundreds or thousands of loci are rare for most invertebrate groups, including freshwater gastropods. This can prevent understanding of phylogeny, which hinders many areas of research. Pleuroceridae is a family of freshwater snails that is highly imperiled and plays an essential role in the ecology of many freshwater systems of the eastern United States. However, the evolutionary history of the family is not understood, and the systematics of the family has not been revised in a modern framework. Pleurocerids display a variety of egg-deposition behaviors and shell shapes, making the family an ideal system for studying evolution of invertebrate life history and morphology. However, past mitochondrial-based phylogenetic analyses have failed to produce meaningful phylogenetic hypotheses, preventing conclusions about pleurocerid systematics and evolution. Here, we generated a novel anchored hybrid enrichment probe set with phylogenetic utility for Pleuroceridae. We sampled pleurocerids from across their range to test the probe set and generated a backbone phylogeny. Our analyses uncovered striking levels of polyphyly among currently accepted genera. Numerous species were also polyphyletic, indicative of unrecognized diversity. Phylogenetic patterns also revealed considerable convergence of shell morphologies. In contrast, anatomical and life history features appeared to be much less homoplastic. Despite generic paraphyly, high support for most major clades and phylogenetic cohesiveness of non-shell characters indicate utility of the AHE probe set for studying pleurocerid evolution

Differential DNA accessibility to polymerase enables 30-minute phenotypic β-lactam antibiotic susceptibility testing of carbapenem-resistant Enterobacteriaceae

The rise in carbapenem-resistant Enterobacteriaceae (CRE) infections has created a global health emergency, underlining the critical need to develop faster diagnostics to treat swiftly and correctly. Although rapid pathogen-identification (ID) tests are being developed, gold-standard antibiotic susceptibility testing (AST) remains unacceptably slow (1–2 d), and innovative approaches for rapid phenotypic ASTs for CREs are urgently needed. Motivated by this need, in this manuscript we tested the hypothesis that upon treatment with β-lactam antibiotics, susceptible Enterobacteriaceae isolates would become sufficiently permeabilized, making some of their DNA accessible to added polymerase and primers. Further, we hypothesized that this accessible DNA would be detectable directly by isothermal amplification methods that do not fully lyse bacterial cells. We build on these results to develop the polymerase-accessibility AST (pol-aAST), a new phenotypic approach for β-lactams, the major antibiotic class for gram-negative infections. We test isolates of the 3 causative pathogens of CRE infections using ceftriaxone (CRO), ertapenem (ETP), and meropenem (MEM) and demonstrate agreement with gold-standard AST. Importantly, pol-aAST correctly categorized resistant isolates that are undetectable by current genotypic methods (negative for β-lactamase genes or lacking predictive genotypes). We also test contrived and clinical urine samples. We show that the pol-aAST can be performed in 30 min sample-to-answer using contrived urine samples and has the potential to be performed directly on clinical urine specimens

Community and service provider views to inform the 2013 WHO consolidated antiretroviral guidelines:key findings and lessons learnt

Objective:The objective was to evaluate community and healthcare worker (HCW) values and preferences on key topics to inform the development of the 2013 WHO consolidated guidelines for antiretroviral therapy in low and middle income countries. Design:Cross-sectional e-survey and e-forum discussion; focus group discussions (FGDs) Methods:Data were collected on community perspectives regarding a range of potential clinical and operational recommendations in the 2013 guidelines between November 2012 and January 2013 through an e-survey (n = 1088) and e-forum (n = 955). Additional FGDs were held with people living with HIV (PLHIV) in Malawi and Uganda (n = 88) on antiretroviral therapy (ART) use among pregnant women. Two surveys were also undertaken on similar topics covered in the e-survey for health care workers caring for adults (n = 98) and children (n = 348). Results:There were 1088 e-survey respondents from 117 countries: of whom 37.7% (298/791) were females, 49.9% (431/864) PLHIV, and 20.9% (174/831) from low-income countries. The proportion of e-survey respondents who supported raising the CD4 T-cell threshold for ART initiation in adults from 350 to 500 cells/μl was 51.0% (355/696), and regardless of CD4 T-cell count for all pregnant females 89.8% (607/676), HIV serodiscordant partners 71.9% (486/676), and all children on diagnosis of infection 47.4% (212/447). E-survey respondents strongly supported discontinuing use of stavudine (72.7%, 416/572), task-shifting/sharing from doctors to nurses (75.2%, 275/365) and from nurses to community health workers (71.1%, 261/367) as strategies to expand access to HIV testing, care, and treatment. Focus group discussion respondents identified service capacity, and social and legal concerns as key considerations influencing the decisions of women living with HIV to continue ART after the risk of vertical transmission has passed. Key lessons learnt in these consultations included the need for piloting and validation of questions; sufficient time to adequately disseminate the survey; and consideration of using FGDs and mobile phone technology to improve participation of people with limited internet access. Conclusion:Community participation in guideline development processes is important to ensure that their perspectives are considered in the resulting recommendations. Communities should be actively involved in the adaptation, implementation, and accountability processes related to the guidelines

Religion and HIV in Tanzania: Influence of Religious Beliefs on HIV stigma, Disclosure, and Treatment Attitudes.

Religion shapes everyday beliefs and activities, but few studies have examined its associations with attitudes about HIV. This exploratory study in Tanzania probed associations between religious beliefs and HIV stigma, disclosure, and attitudes toward antiretroviral (ARV) treatment. A self-administered survey was distributed to a convenience sample of parishioners (n = 438) attending Catholic, Lutheran, and Pentecostal churches in both urban and rural areas. The survey included questions about religious beliefs, opinions about HIV, and knowledge and attitudes about ARVs. Multivariate logistic regression analysis was performed to assess how religion was associated with perceptions about HIV, HIV treatment, and people living with HIV/AIDS. Results indicate that shame-related HIV stigma is strongly associated with religious beliefs such as the belief that HIV is a punishment from God (p < 0.01) or that people living with HIV/AIDS (PLWHA) have not followed the Word of God (p < 0.001). Most participants (84.2%) said that they would disclose their HIV status to their pastor or congregation if they became infected. Although the majority of respondents (80.8%) believed that prayer could cure HIV, almost all (93.7%) said that they would begin ARV treatment if they became HIV-infected. The multivariate analysis found that respondents' hypothetical willingness to begin ARV treatme was not significantly associated with the belief that prayer could cure HIV or with other religious factors. Refusal of ARV treatment was instead correlated with lack of secondary schooling and lack of knowledge about ARVs. The decision to start ARVs hinged primarily on education-level and knowledge about ARVs rather than on religious factors. Research results highlight the influence of religious beliefs on HIV-related stigma and willingness to disclose, and should help to inform HIV-education outreach for religious groups

The influence of variations in eating disorder-related symptoms on processing of emotional faces in a non-clinical female sample:an eye-tracking study

This study aimed to: i) determine if the attention bias towards angry faces reported in eating disorders generalises to a non-clinical sample varying in eating disorder-related symptoms; ii) examine if the bias occurs during initial orientation or later strategic processing; and iii) confirm previous findings of impaired facial emotion recognition in non-clinical disordered eating. Fifty-two females viewed a series of face-pairs (happy or angry paired with neutral) whilst their attentional deployment was continuously monitored using an eye-tracker. They subsequently identified the emotion portrayed in a separate series of faces. The highest (n=18) and lowest scorers (n=17) on the Eating Disorders Inventory (EDI) were compared on the attention and facial emotion recognition tasks. Those with relatively high scores exhibited impaired facial emotion recognition, confirming previous findings in similar non-clinical samples. They also displayed biased attention away from emotional faces during later strategic processing, which is consistent with previously observed impairments in clinical samples. These differences were related to drive-for-thinness. Although we found no evidence of a bias towards angry faces, it is plausible that the observed impairments in emotion recognition and avoidance of emotional faces could disrupt social functioning and act as a risk factor for the development of eating disorders