Reprogramming homing endonuclease specificity through computational design and directed evolution.

Homing endonucleases (HEs) can be used to induce targeted genome modification to reduce the fitness of pathogen vectors such as the malaria-transmitting Anopheles gambiae and to correct deleterious mutations in genetic diseases. We describe the creation of an extensive set of HE variants with novel DNA cleavage specificities using an integrated experimental and computational approach. Using computational modeling and an improved selection strategy, which optimizes specificity in addition to activity, we engineered an endonuclease to cleave in a gene associated with Anopheles sterility and another to cleave near a mutation that causes pyruvate kinase deficiency. In the course of this work we observed unanticipated context-dependence between bases which will need to be mechanistically understood for reprogramming of specificity to succeed more generally.

Comparison of body mass index with waist circumference and skinfold-based percent body fat in firefighters: adiposity classification and associations with cardiovascular disease risk factors.

PURPOSE: This study aims to examine whether body mass index (BMI) overestimates the prevalence of overweight or obese firefighters when compared to waist circumference (WC) and skinfold-based percent body fat (PBF) and to investigate differential relationships of the three adiposity measures with other biological cardiovascular disease (CVD) risk factors. METHODS: The adiposity of 355 (347 males and eight females) California firefighters was assessed using three different measures. Other CVD risk factors (high blood pressure, high lipid profiles, high glucose, and low VO2 max) of the firefighters were also clinically assessed. RESULTS: The prevalence of total overweight and obesity was significantly (p < 0.01) higher by BMI (80.4 %) than by WC (48.7 %) and by PBF (55.6 %) in male firefighters. In particular, the prevalence of overweight firefighters was much higher (p < 0.01) by BMI (57.3 %) than by WC (24.5 %) and PBF (38.3 %). 60-64 % of male firefighters who were assessed as normal weight by WC and PBF were misclassified as overweight by BMI. When overweight by BMI was defined as 27.5-29.9 kg/m(2) (vs. the standard definition of 25.0-29.9 kg/m(2)), the agreement of the adiposity classification increased between BMI and other two adiposity measures. Obese firefighters had the highest CVD risk profiles across all three adiposity measures. Only when overweight by BMI was defined narrowly, overweight firefighters had substantially higher CVD risk profiles. Obesity and overweight were less prevalent in female and Asian male firefighters. CONCLUSIONS: BMI overestimated the prevalence of total overweight and obesity among male firefighters, compared to WC and skinfold-based PBF. Overweight by BMI needs to be more narrowly defined, or the prevalence of BMI-based overweight (27.5-29.9 kg/m(2)) should be reported additionally for prevention of CVD among male firefighters.

Workplace psychosocial and organizational factors for neck pain in workers in the United States.

BACKGROUND: Neck pain is a prevalent musculoskeletal condition among workers in the United States. This study explores a set of workplace psychosocial and organization-related factors for neck pain. METHODS: Data used for this study come from the 2010 National Health Interview Survey which provides a representative sample of the US population. To account for the complex sampling design, the Taylor linearized variance estimation method was used. Logistic regression models were constructed to measure the associations. RESULTS: This study demonstrated significant associations between neck pain and a set of workplace risk factors, including work-family imbalance, exposure to a hostile work environment and job insecurity, non-standard work arrangements, multiple jobs, and long work hours. CONCLUSION: Workers with neck pain may benefit from intervention programs that address issues related to these workplace risk factors. Future studies exploring both psychosocial risk factors and physical risk factors with a longitudinal design will be important. Am. J. Ind. Med. 59:549-560, 2016. © 2016 Wiley Periodicals, Inc.

Low Back Pain Prevalence and Related Workplace Psychosocial Risk Factors: A Study Using Data From the 2010 National Health Interview Survey.

OBJECTIVES: The objectives of this study were to estimate prevalence of low back pain, to investigate associations between low back pain and a set of emerging workplace risk factors, and to identify worker groups with an increased vulnerability for low back pain in the United States. METHODS: The data used for this cross-sectional study came from the 2010 National Health Interview Survey, which was designed to collect data on health conditions and related risk factors from the US civilian population. The variance estimation method was used to compute weighted data for prevalence of low back pain. Multivariable logistic regression analyses stratified by sex and age were performed to determine the odds ratios (ORs) and the 95% confidence interval (CI) for low back pain. The examined work-related psychosocial risk factors included work-family imbalance, exposure to a hostile work environment, and job insecurity. Work hours, occupation, and other work organizational factors (nonstandard work arrangements and alternative shifts) were also examined. RESULTS: The prevalence of self-reported low back pain in the previous 3 months among workers in the United States was 25.7% in 2010. Female or older workers were at increased risk of experiencing low back pain. We found significant associations between low back pain and a set of psychosocial factors, including work-family imbalance (OR 1.27, CI 1.15-1.41), exposure to hostile work (OR 1.39, CI 1.25-1.55), and job insecurity (OR 1.44, CI 1.24-1.67), while controlling for demographic characteristics and other health-related factors. Older workers who had nonstandard work arrangements were more likely to report low back pain. Women who worked 41 to 45 hours per week and younger workers who worked >60 hours per week had an increased risk for low back pain. Workers from several occupation groups, including male health care practitioners, female and younger health care support workers, and female farming, fishing, and forestry workers, had an increased risk of low back pain. CONCLUSIONS: This study linked low back pain to work-family imbalance, exposure to a hostile work environment, job insecurity, long work hours, and certain occupation groups. These factors should be considered by employers, policymakers, and health care practitioners who are concerned about the impact of low back pain in workers.

THE AFFORDABLE CARE ACT: A HIDDEN JOBS KILLER?

Many opponents of the Affordable Care Act (ACA) predicted the law would hurt employment. A common complaint was that the requirement that firms employing more than 50 employees either provide insurance for employees working more than 30 hours a week or pay a penalty would lead many firms to cut back workers' hours to just below the 30-hour cutoff. This article uses the Current Population Survey to compare the share of workers employed between 25-29 hours in the first four months of 2013 with the same months of 2012. While the Obama administration subsequently delayed the application of this portion of the ACA, at the time larger employers would have expected to be subject to the insurance requirement. The analysis found the share of employment in this hour band was slightly lower in 2013 than in 2012.

Recruitment of women in the National Children's Study Initial Vanguard Study.

The initial Vanguard Study of the National Children's Study was conducted during 2009-2010 in 7 locations in the United States. A goal was to evaluate the feasibility and yield of a household-based sampling design to recruit pregnant women. A multistage area probability sampling design was used to identify study locations (generally, counties) that were subsequently divided into smaller geographical units, termed segments. Between 7 and 18 segments were selected in each location, and dwelling units within segments were listed. A household-based recruitment process was implemented, which included enumeration of households to identify age-eligible women, pregnancy screening to identify pregnant women eligible for immediate enrollment and nonpregnant women for telephone follow-up, and administration of informed consent to eligible women. After a recruitment period of 17-20 months, 67,181 (89%) households were enumerated, which identified 34,172 (88%) age-eligible women to whom the pregnancy screener was administered. Among those who completed the screener, 2,285 women became eligible for enrollment, of whom 1,399 (61%) enrolled. Although response rates were fairly high at initial contact and among pregnant women, the overall yield was lower than anticipated. In particular, telephone follow-up of nonpregnant women was not a practicable strategy for prospective recruitment of newly pregnant women.

Regulation of Anopheles gambiae male accessory gland genes influences postmating response in female.

In Drosophila, the accessory gland proteins (Acps) secreted from the male accessory glands (MAGs) and transferred along with sperm into the female reproductive tract have been implicated in triggering postmating behavioral changes, including refractoriness to subsequent mating and propensity to egg laying. Recently, Acps have been found also in Anopheles, suggesting similar functions. Understanding the mechanisms underlying transcriptional regulation of Acps and their functional role in modulating Anopheles postmating behavior may lead to the identification of novel vector control strategies to reduce mosquito populations. We identified heat-shock factor (HSF) binding sites within the Acp promoters of male Anopheles gambiae and discovered three distinct Hsf isoforms; one being significantly up-regulated in the MAGs after mating. Through genome-wide transcription analysis of Hsf-silenced males, we observed significant down-regulation in 50% of the Acp genes if compared to control males treated with a construct directed against an unrelated bacterial sequence. Treated males retained normal life span and reproductive behavior compared to control males. However, mated wild-type females showed a ∼46% reduction of egg deposition rate and a ∼23% reduction of hatching rate (∼58% combined reduction of progeny). Our results highlight an unsuspected role of HSF in regulating Acp transcription in A. gambiae and provide evidence that Acp down-regulation in males leads a significant reduction of progeny, thus opening new avenues toward the development of novel vector control strategies.

Short-term test-retest reliability of the Job Content Questionnaire and Effort-Reward Imbalance Questionnaire items and scales among professional firefighters.

Relatively little is known about the short-term test-retest reliability of the Job Content Questionnaire (JCQ) and Effort-Reward Imbalance Questionnaire (ERIQ). Seventeen JCQ and six ERIQ items were qualitatively reviewed by 19 firefighters in focus groups. The items were then administered twice to 81 firefighters with a time interval of 1-8 weeks. The short-term reliability of the JCQ and ERIQ items and scales with the four-point Likert item responses was at least fair or moderate with several complementary statistical methods. It improved substantially when the four-point responses were simplified into the two ('agree' or 'disagree') responses. The JCQ psychological demands and the ERIQ effort scales were among the least reliable scales and their items were most frequently indicated by the firefighters to be clarified. The responses of professional firefighters to the JCQ and ERIQ items and scales were stable during an 8-week period, particularly when dichotomous item responses were used.

Increases in New but Not Returning Families to a Midwestern Food Pantry Network During the COVID-19 Pandemic.

BACKGROUND: Food pantries are a resource for those experiencing food insecurity. OBJECTIVE: The purpose of this study was to evaluate changes in food pantry utilization and volunteerism for a food pantry network during the COVID-19 pandemic. DESIGN: This 36-month longitudinal, observational study is a secondary analysis of data collected from an Ohio food pantry network. PARTICIPANTS/SETTING: Participants were clients of a food pantry network. The data represent visits from 12 months prepandemic (March 2019 through February 2020), during the pandemic (March 2020 through February 2021), and after vaccines were readily available (March 2021 through February 2022). MAIN OUTCOME MEASURES: Each data point represents a visit to the pantry network. The main outcome measures were total, returning, and new visits (ie, households that had not previously used this pantry network). The secondary outcome was volunteer hours by month. STATISTICAL ANALYSIS: In order to account for month-to-month variability in pantry utilization, the data were analyzed using a time-series linear regression analysis with the month as the unit of analysis. RESULTS: A total of 174,397 visits were recorded during the course of 36 months. Sixty-nine percent of reporting household members were female, 48% reported at least 1 senior in the home, and 41% reported at least 1 child at home. There was no significant change in total or returning visits during the pandemic or after vaccines were available compared with prepandemic levels. However, there was a significant increase in new households to the food pantry network during the pandemic compared with prepandemic (P = .05). Volunteer hours decreased significantly during the pandemic compared with prepandemic months and remained low even after vaccines were available (P = .004 and P = .003, respectively). CONCLUSIONS: Although there was an increase in households new to the food pantry, overall utilization did not increase.

New models for large prospective studies: is there a risk of throwing out the baby with the bathwater?

Manolio et al. (Am J Epidemiol. 2012;175:859-866) proposed that large cohort studies adopt novel models using "temporary assessment centers" to enroll up to a million participants to answer research questions about rare diseases and "harmonize" clinical endpoints collected from administrative records. Extreme selection bias, we are told, will not harm internal validity, and "process expertise to maximize efficiency of high-throughput operations is as important as scientific rigor" (p. 861). In this article, we describe serious deficiencies in this model as applied to the United States. Key points include: 1) the need for more, not less, specification of disease endpoints; 2) the limited utility of data collected from existing administrative and clinical databases; and 3) the value of university-based centers in providing scientific expertise and achieving high recruitment and retention rates through community and healthcare provider engagement. Careful definition of sampling frames and high response rates are crucial to avoid bias and ensure inclusion of important subpopulations, especially the medically underserved. Prospective hypotheses are essential to refine study design, determine sample size, develop pertinent data collection protocols, and achieve alliances with participants and communities. It is premature to reject the strengths of large national cohort studies in favor of a new model for which evidence of efficiency is insufficient.

Maternal perspectives on the return of genetic results: context matters.

The objectives of this study were to study maternal preferences for the return of their child's genetic results and to describe the experiences, perceptions, attitudes, and values that are brought to bear when individuals from different racial and cultural backgrounds consider participating in genetic research. We recruited women with diverse sociodemographic profiles to participate in seven focus groups. Twenty-eight percent of participants self-identified as Hispanic; 49% as White, non-Hispanic; and 21% as Asian or Asian American. Focus groups were conducted in English or Spanish and were audio-recorded and transcribed verbatim. Transcripts were analyzed using qualitative thematic methods. Results indicated that preferences and decisions regarding the return of results may depend on both research and individual contextual factors. Participants understood the return of results as a complex issue, where individual and cultural differences in preferences are certain to arise. Another key finding was that participants desired an interpersonal, dynamic, flexible process that accommodated individual preferences and contextual differences for returning results. Our findings indicate a need to have well-developed systems for allowing participants to make and change over time their choices regarding the return of their child's genetic results.

Effects of gestational and lactational exposure to heptachlor epoxide on age at puberty and reproductive function in men and women.

BACKGROUND: For 15 months in 1981-1982, the commercial milk supply on the Hawaiian island of Oahu was contaminated with heptachlor epoxide, a metabolite of the insecticide heptachlor, resulting in gestational and/or lactational exposure to offspring of women who drank cow milk during that period. OBJECTIVE: To determine whether gestational and lactational exposure to heptachlor epoxide alters reproductive function and age at puberty in men or women. METHODS: 457 participants were recruited from a prior high school enrollment sampling frame of 20,000 adults born during 1981-1982 who lived on Oahu since at least first grade. Number of glasses of cow milk consumed weekly by the mother during the participant's gestation was used as a surrogate measure of heptachlor epoxide exposure. Reproductive function measures included semen analyses; reproductive hormones or their metabolites in daily urine specimens for one menstrual cycle; serum reproductive hormone levels in both sexes; and reported ages of onset for pubertal milestones. RESULTS: We observed no strong associations of heptachlor epoxide exposure during gestation and lactation with reproductive endpoints. In females, heptachlor epoxide exposure was associated with longer luteal phase length and slower drop in the ratio of estradiol to progesterone metabolites after ovulation. In males, heptachlor epoxide exposure was weakly associated with higher serum follicle stimulating hormone and luteinizing hormone concentrations, but no dose-response relationship was apparent. CONCLUSIONS: The results provide limited evidence that gestational and lactational exposure to heptachlor epoxide, due to milk contamination on Oahu in 1981-1982, resulted in clinically significant disturbances of reproductive function in men or women.

Travel patterns during pregnancy: comparison between Global Positioning System (GPS) tracking and questionnaire data.

BACKGROUND: Maternal exposures to traffic-related air pollution have been associated with adverse pregnancy outcomes. Exposures to traffic-related air pollutants are strongly influenced by time spent near traffic. However, little is known about women's travel activities during pregnancy and whether questionnaire-based data can provide reliable information on travel patterns during pregnancy. OBJECTIVES: Examine women's in-vehicle travel behavior during pregnancy and examine the difference in travel data collected by questionnaire and global positioning system (GPS) and their potential for exposure error. METHODS: We measured work-related travel patterns in 56 pregnant women using a questionnaire and one-week GPS tracking three times during pregnancy (<20 weeks, 20-30 weeks, and >30 weeks of gestation). We compared self-reported activities with GPS-derived trip distance and duration, and examined potentially influential factors that may contribute to differences. We also described in-vehicle travel behavior by pregnancy periods and influences of demographic and personal factors on daily travel times. Finally, we estimated personal exposure to particle-bound polycyclic aromatic hydrocarbon (PB-PAH) and examined the magnitude of exposure misclassification using self-reported vs. GPS travel data. RESULTS: Subjects overestimated both trip duration and trip distance compared to the GPS data. We observed moderately high correlations between self-reported and GPS-recorded travel distance (home to work trips: r = 0.88; work to home trips: r = 0.80). Better agreement was observed between the GPS and the self-reported travel time for home to work trips (r = 0.77) than work to home trips (r = 0.64). The subjects on average spent 69 and 93 minutes traveling in vehicles daily based on the GPS and self-reported data, respectively. Longer daily travel time was observed among participants in early pregnancy, and during certain pregnancy periods in women with higher education attainment, higher income, and no children. When comparing self-reported vs. GPS data, we found that estimated personal exposure to PB-PAH did not differ remarkably at the population level, but the difference was large at an individual level. CONCLUSION: Self-reported home-to-work data overestimated both trip duration and trip distance compared to GPS data. Significant differences in PAH exposure estimates were observed at individual level using self-reported vs. GPS data, which has important implications in air pollution epidemiological studies.

Exploring occupational and health behavioral causes of firefighter obesity: a qualitative study.

BACKGROUND: Firefighters, as an occupational group, have one of the highest prevalence rates of obesity. A qualitative study investigated occupational and health behavioral determinants of obesity among firefighters. METHODS: Four focus groups were conducted with firefighters of every rank as Phase I of the FORWARD study which was designed to assess health behavioral and occupational characteristics related to obesity in firefighters. RESULTS: Analysis revealed five main themes of central importance to firefighters: (1) fire station eating culture; (2) night calls and sleep interruption; (3) supervisor leadership and physical fitness; (4) sedentary work; and (5) age and generational influences. CONCLUSION: The results showed a strong interrelationship between occupational and health behavioral causes of obesity in firefighters. The relevance of these qualitative findings are discussed along with the implications for future obesity interventions with firefighters.

Simple Neurite Tracer: open source software for reconstruction, visualization and analysis of neuronal processes.

MOTIVATION: Advances in techniques to sparsely label neurons unlock the potential to reconstruct connectivity from 3D image stacks acquired by light microscopy. We present an application for semi-automated tracing of neurons to quickly annotate noisy datasets and construct complex neuronal topologies, which we call the Simple Neurite Tracer. AVAILABILITY: Simple Neurite Tracer is open source software, licensed under the GNU General Public Licence (GPL) and based on the public domain image processing software ImageJ. The software and further documentation are available via http://fiji.sc/Simple_Neurite_Tracer as part of the package Fiji, and can be used on Windows, Mac OS and Linux. Documentation and introductory screencasts are available at the same URL. CONTACT: longair@ini.phys.ethz.ch; longair@ini.phys.ethz.ch.

Blood cadmium and moderate-to-severe glomerular dysfunction in Korean adults: analysis of KNHANES 2005-2008 data.

PURPOSE: The objective of this study was to evaluate the association between blood cadmium (Cd) and moderate-to-severe glomerular dysfunction in a Korean population using a representative sample. METHODS: A cross-sectional study was used to evaluate the association between blood Cd and glomerular dysfunction. Based on the Korean National Health and Nutrition Examination Survey (2005-2008), individual blood Cd was measured in 2,992 adults, aged between 20 and 65. RESULTS: After adjusting for survey years, age, sociodemographic factors, and health behaviors, the odds ratio for moderate-to-severe glomerular dysfunction (<60 mL/min per 1.73 m(2)) was 1.97 (95% CI: 1.28-3.07) when comparing the highest with the lowest blood Cd quartile in Korean women. However, in Korean men, there was no association between blood Cd and moderate-to-severe glomerular dysfunction. CONCLUSIONS: These findings support the consideration of Cd as a risk factor for glomerular dysfunction in the female population. Furthermore, environmental heavy metal monitoring and an institutional strategy should be implemented to reduce Cd exposure in the general population.

Diverse perceptions of the informed consent process: implications for the recruitment and participation of diverse communities in the National Children's Study.

We examined the experiences, perceptions, and values that are brought to bear when individuals from different ethnic and cultural backgrounds consider participating in health research. Fifty-three women from Latino, Asian American, Middle Eastern, or Non-Latino, White backgrounds participated in seven English or Spanish focus groups facilitated by trained investigators using a standard protocol. Investigators described the National Children's Study (NCS) and then asked questions to elicit potential concerns, expectations, and informational needs. Group sessions were audio-recorded, transcribed verbatim, and analyzed using qualitative thematic methods. A major theme that emerged during focus groups was participant self-identification as a member of a cultural group or community when raising issues that would influence their decision to participate in research. A related theme was the belief by some that communities may differ in the ease of participation in the NCS. Identified themes related to the informed consent process included perceived risks, anticipated burden, perceived benefits, informational needs, and decision-making strategies. Although themes were shared across groups, there were cultural differences within themes. Findings indicated that individuals from diverse backgrounds may have different perspectives on and expectations for the research process. To effectively recruit representative samples, it will be important to address a range of issues relevant for informed consent and to consider the impact of participation on both individuals and communities.

Occupation-differential construct validity of the Job Content Questionnaire (JCQ) psychological job demands scale with physical job demands items: a mixed methods research.

The construct validity of the Job Content Questionnaire (JCQ) psychological demands scale in relationship to physical demands has been inconsistent. This study aims to test quantitatively and qualitatively whether the scale validity differs by occupation. Hierarchical clustering analyses of 10 JCQ psychological and physical demands items were conducted in 61 occupations from two datasets: one of non-faculty workers at a university in the United States (6 occupations with 208 total workers) and the other of a Belgian working population (55 occupations with 13,039 total workers). The psychological and physical demands items overlapped in 13 of 61 occupation-stratified clustering analyses. Most of the overlaps occurred in physically-demanding occupations and involved the two psychological demands items, 'work fast' and 'work hard'. Generally, the scale reliability was low in such occupations. Additionally, interviews with eight university workers revealed that workers interpreted the two psychological demands items differently by the nature of their tasks. The scale validity was occupation-differential. PRACTITIONER SUMMARY: The JCQ psychological job demands scale as a job demand measure has been used worldwide in many studies. This study indicates that the wordings of the 'work fast' and 'work hard' items of the scale need to be reworded enough to differentiate mental and physical job demands as intended, 'psychological.'

A comprehensive gene expression atlas of sex- and tissue-specificity in the malaria vector, Anopheles gambiae.

BACKGROUND: The mosquito, Anopheles gambiae, is the primary vector of human malaria, a disease responsible for millions of deaths each year. To improve strategies for controlling transmission of the causative parasite, Plasmodium falciparum, we require a thorough understanding of the developmental mechanisms, physiological processes and evolutionary pressures affecting life-history traits in the mosquito. Identifying genes expressed in particular tissues or involved in specific biological processes is an essential part of this process. RESULTS: In this study, we present transcription profiles for ~82% of annotated Anopheles genes in dissected adult male and female tissues. The sensitivity afforded by examining dissected tissues found gene activity in an additional 20% of the genome that is undetected when using whole-animal samples. The somatic and reproductive tissues we examined each displayed patterns of sexually dimorphic and tissue-specific expression. By comparing expression profiles with Drosophila melanogaster we also assessed which genes are well conserved within the Diptera versus those that are more recently evolved. CONCLUSIONS: Our expression atlas and associated publicly available database, the MozAtlas (http://www.tissue-atlas.org), provides information on the relative strength and specificity of gene expression in several somatic and reproductive tissues, isolated from a single strain grown under uniform conditions. The data will serve as a reference for other mosquito researchers by providing a simple method for identifying where genes are expressed in the adult, however, in addition our resource will also provide insights into the evolutionary diversity associated with gene expression levels among species.