Nesting Behavior of Palila, as Assessed from Video Recordings

We quantified nesting behavior of Palila (Loxioides bailleui), an endangered Hawaiian honeycreeper, by recording at nests during three breeding seasons using a black-and-white video camera connected to a videocassette recorder. A total of seven nests was observed. We measured the following factors for daylight hours: percentage of time the female was on the nest (attendance), length of attendance bouts by the female, length of nest recesses, and adult provisioning rates. Comparisons were made between three stages of the 40-day nesting cycle: incubation (day 1–day 16), early nestling stage (day 17–day 30 [i.e., nestlings ≤ 14 days old]), and late nestling stage (day 31–day 40 [i.e., nestlings \u3e 14 days old]). Of seven nests observed, four fledged at least one nestling and three failed. One of these failed nests was filmed being depredated by a feral cat (Felis catus). Female nest attendance was near 82% during the incubation stage and decreased to 21% as nestlings aged. We did not detect a difference in attendance bout length between stages of the nesting cycle. Mean length of nest recesses increased from 4.5 min during the incubation stage to over 45 min during the late nestling stage. Mean number of nest recesses per hour ranged from 1.6 to 2.0. Food was delivered to nestlings by adults an average of 1.8 times per hour for the early nestling stage and 1.5 times per hour during the late nestling stage and did not change over time. Characterization of parental behavior by video had similarities to but also key differences from findings taken from blind observations. Results from this study will facilitate greater understanding of Palila reproductive strategies

Quantifying and resolving multiple vector transformants in S. cerevisiae plasmid libraries

<p>Abstract</p> <p>Background</p> <p>In addition to providing the molecular machinery for transcription and translation, recombinant microbial expression hosts maintain the critical genotype-phenotype link that is essential for high throughput screening and recovery of proteins encoded by plasmid libraries. It is known that <it>Escherichia coli </it>cells can be simultaneously transformed with multiple unique plasmids and thusly complicate recombinant library screening experiments. As a result of their potential to yield misleading results, bacterial multiple vector transformants have been thoroughly characterized in previous model studies. In contrast to bacterial systems, there is little quantitative information available regarding multiple vector transformants in yeast. <it>Saccharomyces cerevisiae </it>is the most widely used eukaryotic platform for cell surface display, combinatorial protein engineering, and other recombinant library screens. In order to characterize the extent and nature of multiple vector transformants in this important host, plasmid-born gene libraries constructed by yeast homologous recombination were analyzed by DNA sequencing.</p> <p>Results</p> <p>It was found that up to 90% of clones in yeast homologous recombination libraries may be multiple vector transformants, that on average these clones bear four or more unique mutant genes, and that these multiple vector cells persist as a significant proportion of library populations for greater than 24 hours during liquid outgrowth. Both vector concentration and vector to insert ratio influenced the library proportion of multiple vector transformants, but their population frequency was independent of transformation efficiency. Interestingly, the average number of plasmids born by multiple vector transformants did not vary with their library population proportion.</p> <p>Conclusion</p> <p>These results highlight the potential for multiple vector transformants to dominate yeast libraries constructed by homologous recombination. The previously unrecognized prevalence and persistence of multiply transformed yeast cells have important implications for yeast library screens. The quantitative information described herein should increase awareness of this issue, and the rapid sequencing approach developed for these studies should be widely useful for identifying multiple vector transformants and avoiding complications associated with cells that have acquired more than one unique plasmid.</p

Emergency supply of prescription-only medicines to patients by community pharmacists: a mixed methods evaluation incorporating patient, pharmacist and GP perspectives

Objective To evaluate and inform emergency supply of prescription-only medicines by community pharmacists (CPs), including how the service could form an integral component of established healthcare provision to maximise adherence. Design Mixed methods. 4 phases: prospective audit of emergency supply requests for prescribed medicines (October–November 2012 and April 2013); interviews with CPs (February–April 2013); follow-up interviews with patients (April–May 2013); interactive feedback sessions with general practice teams (October–November 2013). Setting 22 community pharmacies and 6 general practices in Northwest England. Participants 27 CPs with experience of dealing with requests for emergency supplies; 25 patients who received an emergency supply of a prescribed medicine; 58 staff at 6 general practices. Results Clinical audit in 22 pharmacies over two 4-week periods reported that 526 medicines were requested by 450 patients. Requests peaked over a bank holiday and around weekends. A significant number of supplies were made during practice opening hours. Most requests were for older patients and for medicines used in long-term conditions. Difficulty in renewing repeat medication (forgetting to order, or prescription delays) was the major reason for requests. The majority of medicines were ‘loaned’ in advance of a National Health Service (NHS) prescription. Interviews with CPs and patients indicated that continuous supply had a positive impact on medicines adherence, removing the need to access urgent care. General practice staff were surprised and concerned by the extent of emergency supply episodes. Conclusions CPs regularly provide emergency supplies to patients who run out of their repeat medication, including during practice opening hours. This may aid adherence. There is currently no feedback loop, however, to general practice. Patient care and interprofessional communication may be better served by the introduction of a formally structured and funded NHS emergency supply service from community pharmacies, with ongoing optimisation of repeat prescribing

Healthy Food Accessibility in Grocery Stores in Central Massachusetts

BACKGROUND. Accessibility to healthy food is one of the most influential community-level factors affecting obesity and chronic disease. The Community Nutrition Environment Evaluation Data System (C-NEEDS) is a set of instruments for objectively assessing availability and quality of 61 major healthy and unhealthy food items in foods stores in the Northeast region. METHODS. The C-NEEDS was developed considering seasonal variations, cultural relevance and utility to cardiovascular health research. Both inter- and intra-rater reliability tests showed a high degree of agreement. Using the instruments, we conducted four rounds of longitudinal surveys of 107 grocery stores in Worcester County, Massachusetts between 2007 and 2010. A healthy food availability index (HFAI, 0-37 points) was calculated for each store, a higher score indicating a greater availability and better quality of healthy foods. Using linear regression models, we examined variations in HFAI in relation to community household income and housing density. RESULTS. Store-level HFAI did not vary significantly by tertile of community median income, but did vary by housing density. High-density communities (upper tertile) had the greatest percentage of stores in the top HFAI tertile (34-37 points). Middle-density communities had the greatest percentage of stores in the low HFAI tertile (0-17 points). A majority of the stores located in low-density communities had middle range of HFAI (18-33 points). The mean HFAI increased with each successive round of grocery store surveys (β=2.02/round [95% confidence interval 0.74-3.31]). CONCLUSION. Access to healthy foods improved slightly over time, however, notable disparities still existed in Central Massachusetts during the study period. Better access was associated with community housing density but not median household income. Further studies on the causes of the disparities may inform public health organizations about necessary community actions to reduce these disparities

Sensing mechanisms of iron–sulfur cluster regulatory proteins elucidated using native mass spectrometry

The ability to sense and respond to various key environmental cues is important for the survival and adaptability of many bacteria, including pathogens. The particular sensitivity of iron–sulfur (Fe–S) clusters is exploited in nature, such that multiple sensor-regulator proteins, which coordinate the detection of analytes with a (in many cases) global transcriptional response, are Fe–S cluster proteins. The fragility and sensitivity of these Fe–S clusters make studying such proteins difficult, and gaining insight of what they sense, and how they sense it and transduce the signal to affect transcription, is a major challenge. While mass spectrometry is very widely used in biological research, it is normally employed under denaturing conditions where non-covalently attached cofactors are lost. However, mass spectrometry under conditions where the protein retains its native structure and, thus, cofactors, is now itself a flourishing field, and the application of such ‘native’ mass spectrometry to study metalloproteins is now relatively widespread. Here we describe recent advances in using native MS to study Fe–S cluster proteins. Through its ability to accurately measure mass changes that reflect chemistry occurring at the cluster, this approach has yielded a remarkable richness of information that is not accessible by other, more traditional techniques

Misclassification of Healthy Eating Based on Food Frequency Questionnaires and 24-hour Dietary Recalls in Older Men and Women

OBJECTIVE: To examine the agreement in nutrient intake and alternative healthy eating indices (AHEI) between a self-administered Food Frequency Questionnaire (FFQ) and 24-hour recall (24HR) measurements of diet by gender among older adults. METHODS: This is a cross-sectional observational study of 105 men and 99 women aged 65 and older living in urban and rural neighborhoods in Worcester County. Participants were queried on diet using both FFQ and 24HR. The healthy eating classification was compared between the two instruments by gender. RESULTS: For men, the mean±SD of AHEI total score was 48.2±12.3 based on FFQ versus 34.7±10.2 based on 24HR. For women, the mean±SD was 47.9±10.1 based on FFQ versus 36.1±10.0 based on 24HR. Using 32 as the cutoff (40% of maximum AHEI score), 9% of men and 7% of women were classified as eating unhealthy based on the FFQ, versus 47% of men and 38% of women based on 24HR. Compared to women, men had larger 24HR to FFQ discrepancies in the nuts and vegetable protein subscore and white/red meat ratio, and smaller discrepancy in alcohol beverages subscore. CONCLUSION: Agreements between FFQ and 24HR-based measures of diet quality were roughly comparable between men and women, though slightly better for women than men. Compared to 24HR, the FFQ tended to underestimate the proportions of older men and women classified as eating unhealthy and misclassified more men than women. Such limitations should be considered when the FFQ is used to study healthy eating in older age

Identification of Metabolites of Trenbolone Acetate in Androgenic Runoff from a Beef Feedlot

Little is known concerning the potential ecological effects of hormonally active substances associated with discharges from animal feeding operations. Trenbolone acetate is a synthetic anabolic steroid that is widely used in the United States to promote growth of beef cattle. Metabolites of trenbolone acetate include the stereoisomers 17α- and 17β-trenbolone, both of which are stable in animal wastes and are relatively potent androgens in fish and mammals. Our purpose in this study was to evaluate the occurrence of 17α- and 17β-trenbolone in a beef cattle feedlot discharge and in river water upstream and downstream from the discharge. In conjunction with that effort, we measured in vitro androgenic activity of the discharge using CV-1 cells that had been transiently cotransfected with human androgen receptor and reporter gene constructs. Samples were collected on nine different occasions during 2002 and 2003. Whole-water samples from the discharge caused a significant androgenic response in the CV-1 cells and contained detectable concentrations of 17α- and 17β-trenbolone. Further work is needed to ascertain the degree to which synthetic androgens such as trenbolone contribute to androgenic activity of feedlot discharges

Association between VTE and antibiotic prophylaxis guideline compliance and patient-reported outcomes after total hip and knee arthroplasty : an observational study

Background: Surgical site infection (SSI) and venous thromboembolism (VTE) are associated with high burden and cost and are considered largely preventable following total knee or hip arthroplasty (TKA, THA). The risk of developing VTE and SSI is reduced when prophylaxis is compliant with evidence-based clinical guidelines. However, the association between VTE and antibiotic prophylaxis clinical guideline compliance and patient-reported outcome measures (PROMs) after THA/TKA is unknown. This study aims to explore whether care that is non-compliant with VTE and antibiotic guideline recommendations is associated with PROMs (Oxford Hip/Knee Score and EQ-5D Index scores) at 90- and 365-days after surgery. Methods: This prospective observational study included high-volume arthroplasty public and private sites and consenting eligible participants undergoing elective primary THA/TKA. We conducted multiple linear regression and linear mixed-effects modelling to explore the associations between non-compliance with VTE and antibiotic guidelines, and PROMs. Results: The sample included 1838 participants. Compliance with VTE and antibiotic guidelines was 35% and 13.2% respectively. In adjusted modelling, non-compliance with VTE guidelines was not associated with 90-day Oxford score (β = − 0.54, standard error [SE] = 0.34, p = 0.112) but was significantly associated with lower (worse) 365-day Oxford score (β = − 0.76, SE = 0.29, p = 0.009), lower EQ-5D Index scores at 90- (β = − 0.02 SE = 0.008, p = 0.011) and 365-days (β = − 0.03, SE = 0.008, p = 0.002). The changes in Oxford and EQ-5D Index scores were not clinically important. Noncompliance with antibiotic guidelines was not associated with either PROM at 90- (Oxford: β = − 0.45, standard error [SE] = 0.47, p = 0.341; EQ-5D: β = − 0.001, SE = 0.011, p = 0.891) or 365-days (Oxford score: β = − 0.06, SE = 0.41, p = 0.880 EQ-5D: β = − 0.010, SE = 0.012, p = 0.383). Results were consistent when complications were included in the model and in linear mixed-effects modelling with the insurance sector as a random effect. Conclusions: Non-compliance with VTE prophylaxis guidelines, but not antibiotic guidelines, is associated with statistically significant but not clinically meaningful differences in Oxford scores and EQ-5D Index scores at 365 days

Racial Differences in Neighborhood Perceptions and their Influences on Physical Activity among Urban Older Women

Background: Proper levels of physical activity (PA) are important to healthy aging. Little is known about racial differences in influences of neighborhood perceptions (NP) on PA and use of neighborhood resources among community-dwelling older women. Materials and methods: In 2014 and 2015, 49 white and 44 black women of age 65 and older living in Washington, DC were queried about their PA, NP, use of neighborhood resources and sociodemographic characteristics. They wore an accelerometer and a Global Positioning System device concurrently for 7 consecutive days. Data were analyzed by race. Results: Compared to Whites, Blacks had lower NP scores (71% positive vs. 77%, p = 0.01), lower mean daily step counts (mean (SD): 3256 (1918) vs. 5457 (2989), p \u3c 0.001), and lower frequencies of all exercise activities combined (19.7 (8.7) vs. 25.2 (11.8) per week, p = 0.01). For both Whites and Blacks, better NPs were associated with more frequent PA both at (p = 0.05) and away from home (p = 0.01). However, better NPs were associated with higher frequencies of exercise activities, moderate-to-high intensity activities, and utilitarian walking for Whites but not Blacks (p \u3c 0.05 for race-perception interaction terms). Conclusions: In an urban setting, older Black women were more likely than older White women to have poor NPs, less PA, and weaker or no association of positive NPs with higher levels of certain PAs. Such substantial racial differences warrant further investigation and consideration in health promotion programs