Impact of α-crystallin protein loss on zebrafish lens development.

The α-crystallin small heat shock proteins contribute to the transparency and refractive properties of the vertebrate eye lens and prevent the protein aggregation that would otherwise produce lens cataracts, the leading cause of human blindness. There are conflicting data in the literature as to what role the α-crystallins may play in early lens development. In this study, we used CRISPR gene editing to produce zebrafish lines with mutations in each of the three α-crystallin genes (cryaa, cryaba and cryabb) to prevent protein production. The absence of each α-crystallin protein was analyzed by mass spectrometry, and lens phenotypes were assessed with differential interference contrast microscopy and histology. Loss of αA-crystallin produced a variety of lens defects with varying severity in larvae at 3 and 4 dpf but little substantial change in normal fiber cell denucleation. Loss of αBa-crystallin produced no substantial lens defects. Our cryabb mutant produced a truncated αBb-crystallin protein and showed no substantial change in lens development. Mutation of each α-crystallin gene did not alter the mRNA levels of the remaining two, suggesting a lack of genetic compensation. These data suggest that αA-crystallin plays some role in lens development, but the range of phenotype severity in null mutants indicates its loss simply increases the chance for defects and that the protein is not essential. Our finding that cryaba and cryabb mutants lack noticeable lens defects is congruent with insubstantial transcript levels for these genes in lens epithelial and fiber cells through five days of development. Future experiments can explore the molecular mechanisms leading to lens defects in cryaa null mutants and the impact of αA-crystallin loss during zebrafish lens aging.

Evaluation and Management of Toxicodendron Dermatitis in the Emergency Department: A Review of Current Practices.

Toxicodendron dermatitis is an underappreciated disease seen in the emergency department. Although self-limiting, symptoms can be distressing and can last for weeks if untreated, particularly with re-exposure. Continuing research has improved our understanding of specific inflammatory markers that are associated with exposure to urushiol-the compound responsible for Toxicodendron dermatitis-although consensus for treatment remains varied and poorly supported. Owing to the lack of recent primary literature on the topic, many providers rely on historical precedent, expert opinion, and personal experience when treating this disease. This article provides a narrative review of the literature currently available on the effects of urushiol on key molecular and cellular functions and the prevention and treatment of Toxicodendron dermatitis.

Hypoxic respiratory distress potentially secondary to phosphorus trifluoride gas exposure.

We herein report a rare, probable exposure of a patient to phosphorus trifluoride gas. The objective of this case report is to highlight the potential exposure to phosphorus trifluoride gas and discuss the best management of it. A 48-year-old worker at a specialty gases laboratory was transported to the community Emergency Department (ED) in respiratory distress, presenting with peripheral cyanosis, an unobtainable oxygen saturation, chocolate-colored blood, and a Glasgow coma scale of 15. A non-rebreather was placed, poison control was contacted, and the patient was administered empiric methylene blue intravenously due to worsening cyanosis and respiratory distress. Upon arrival at the academic facility, the patient was no longer cyanotic and reported improvement of his symptoms. The patient's employer informed staff that a canister of phosphorus trifluoride gas in his workstation was found to be empty but should have been full. It was also discovered that a coworker left work early the same day with similar but milder symptoms. Hyperbaric oxygen therapy was considered; however, the patient was improving on oxygen via non-rebreather, and he had no other indications. Because the patient continued to require supplemental oxygen to maintain their oxygen saturation above 92%, he was admitted to the ICU and treated with prednisone daily for chemical pneumonitis. After 4 days, he successfully transitioned to room air without hypoxia. While exposures such as this do not occur frequently, it is important to maintain a broad differential and treatment plan as we continue to investigate the etiology and best treatment option.

Evaluation of intervention reporting in published emergency medicine clinical trials.

OBJECTIVE: The extent of intervention reporting in emergency medicine journals remains unclear. The primary objective is to assess overall completion of the Template for Intervention Description and Replication (TIDieR) checklist described in emergency medicine randomized clinical trials (RCTs). The secondary outcomes were to (1) compare reporting before and after TIDieR publication; (2) evaluate factors associated with intervention reporting. METHODS: Our cross-sectional study used Google Scholar's metrics to identify seven emergency medicine journals; of which, we randomly sampled 300 articles. Using two PubMed searches, we extracted 150 RCTs before and after publications of TIDieR. Two investigators independently extracted data. The primary analysis to measure overall completion included descriptive statistics for each checklist item. Our secondary analysis used an interrupted time series analysis and generalized estimating equations to determine the effect of TIDieR publication on intervention reporting. RESULTS: Our initial search yielded 635 articles; from which, we randomly sampled 300 articles. We excluded 67 articles, leaving 233 for analysis. The mean number of TIDieR items reported was 5.4 (standard deviation = 1.18). Of the 233 trials, 42.9% provided information about materials, 67% provided intervention procedures, and 99.1% provided intervention delivery. The least reported items were intervention modifications (2.6%), intervention adherence assessment methods (3.4%), and intervention adherence assessment outcomes (2.2%). CONCLUSIONS: The completeness of intervention reporting is suboptimal in emergency medicine journals, necessitating improvement. The current state of adherence could be improved through the combined efforts of journal editors, major editorial organizations, and authors.

Applied Sports Nutrition Support, Dietary Intake and Body Composition Changes of a Female Athlete Completing 26 Marathons in 26 Days: A Case Study.

The aim of this case study is to describe the nutrition practices of a female recreational runner (VO2max 48.9 ml·kg-1·min-1) who completed 26 marathons (42.195 km) in 26 consecutive days. Information relating to the nutritional intake of female runners during multi-day endurance events is extremely limited, yet the number of people participating year-on-year continues to increase. This case study reports the nutrition intervention, dietary intake, body composition changes and performance in the lead-up and during the 26 days. Prior to undertaking the 26 marathon challenge, three consultations were held between the athlete and a sports nutrition advisor; planning and tailoring the general diet and race-specific strategies to the endurance challenge. During the marathons, the mean energy and fluid intake was 1039.7 ± 207.9 kcal (607.1 - 1453.2) and 2.39 ± 0.35 L (1.98 - 3.19). Mean hourly carbohydrate intake was 38.9 g·hr-1. 11 days following the completion of the 26 marathons, body mass had reduced by 4.6 kg and lean body mass increasing by 0.53 kg when compared with 20 days prior. This case study highlights the importance of providing general and event-specific nutrition education when training for such an event. This is particularly prudent for multi-day endurance running events.

Associations between child marriage and reproductive and maternal health outcomes among young married women in Liberia and Sierra Leone: A cross-sectional study.

BACKGROUND: Child marriage has been associated with a range of negative maternal and reproductive health outcomes. This study explored these associations in Liberia and Sierra Leone and examined how child marriage intersected with other measures of social disadvantage. METHODS: Data were derived from 631 and 1,325 married or cohabitating women aged 20-24 interviewed in the 2019-2020 Liberia and 2019 Sierra Leone Demographic and Health Surveys, respectively. Analyses were stratified by country. Regression models examined associations between age at first marriage (<15, 15-17, and 18+ years) and reproductive and maternal health outcomes, as well as interactions between child marriage and measures of social disadvantage. Multivariable regression results were presented as adjusted odds ratios and 95% confidence intervals. RESULTS: Over half of currently married/cohabitating women aged 20-24 in Liberia (52%) and Sierra Leone (54%) married before age 18, and over one in 10 married before age 15. In both countries, after adjusting for other factors, being married before the age of 18 was significantly associated with early fertility, high fertility, and low fertility control. Associations were particularly strong among women who first married before age 15. In Liberia, women who married at age 15-17 had significantly lower odds of skilled attendance at delivery and institutional delivery if they lived in the North Central region. Sierra Leonean women who married before age 15 had lower odds of institutional delivery and lower odds of four or more ANC visits if they lived in the North Western region. CONCLUSION: This study found clear associations between child marriage and negative reproductive health outcomes in Liberia and Sierra Leone, with stronger associations among women married in early adolescence. Child marriage and region of residence intersected to shape young women's access to skilled attendance at birth and institutional delivery. These findings call for further investigation and targeted intervention.

Archaeal viruses, not archaeal phages: an archaeological dig.

Viruses infect members of domains Bacteria, Eukarya, and Archaea. While those infecting domain Eukarya are nearly universally described as "Viruses", those of domain Bacteria, to a substantial extent, instead are called "Bacteriophages," or "Phages." Should the viruses of domain Archaea therefore be dubbed "Archaeal phages," "Archaeal viruses," or some other construct? Here we provide documentation of published, general descriptors of the viruses of domain Archaea. Though at first the term "Phage" or equivalent was used almost exclusively in the archaeal virus literature, there has been a nearly 30-year trend away from this usage, with some persistence of "Phage" to describe "Head-and-tail" archaeal viruses, "Halophage" to describe viruses of halophilic Archaea, use of "Prophage" rather than "Provirus," and so forth. We speculate on the root of the early 1980's transition from "Phage" to "Virus" to describe these infectious agents, consider the timing of introduction of "Archaeal virus" (which can be viewed as analogous to "Bacterial virus"), identify numerous proposed alternatives to "Archaeal virus," and also provide discussion of the general merits of the term, "Phage." Altogether we identify in excess of one dozen variations on how the viruses of domain Archaea are described, and document the timing of both their introduction and use.

Planning engagement with web resources to improve diet quality and break up sedentary time for home-working employees: A mixed methods study.

As home working becomes more common, employers may struggle to provide health promotion interventions that can successfully bridge the gap between employees' intentions to engage in healthier behaviors and actual action. Based on past evidence that action planning can successfully encourage the adoption of healthier behaviors, this mixed-methods study of a web-based self-help intervention incorporated a randomized planning trial that included quantitative measures of engagement and follow-up qualitative interviews with a subsample of participants. Participants either (a) selected a movement plan for incorporating a series of 2-min exercise videos into their work week to break up sedentary time and a balanced meal plan with recipe cards for a week's lunches and dinners or (b) received access to these resources without a plan. Selecting a movement plan was more effective at increasing engagement with the web resources compared to the no-plan condition. In the follow-up interviews, participants indicated that the plan helped to remind participants to engage with the resources and made it simpler for them to follow the guidance for exercises and meals. Ease of use and being able to fit exercises and meals around work tasks were key factors that facilitated uptake of the resources, while lack of time and worries about how colleagues would perceive them taking breaks to use the resources were barriers to uptake. Participants' self-efficacy was associated with general resource use but not plan adherence. Overall, including plans with online self-help resources could enhance their uptake. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

The Enneagram and its application in medical education.

Personality testing has become increasingly popular in healthcare with multiple modalities and implementations. Although personality testing has been utilized to inform various facets of graduate medical education, little is known about how the Enneagram can be utilized throughout postgraduate training. This narrative review explores the use of personality testing in graduate medical education, how personality testing has been used in the workplace, what research is available showing its use in medical residencies, and the need for additional studies on the Enneagram's use in these areas. We conclude the Enneagram may serve as a valuable tool that can be used in postgraduate medical education to improve learning, interpersonal relationships, and teaming.

The feasibility, safety, and efficacy of lower limb garment-integrated blood flow restriction training in healthy adults.

OBJECTIVES: Explore the feasibility of lower-limb garment-integrated BFR-training. DESIGN: Observational study. SETTING: Human performance laboratory. PARTICIPANTS: Healthy males with no experience of BFR-training. MAIN OUTCOME MEASURES: Feasibility was determined by a priori thresholds for recruitment, adherence, and data collection. Safety was determined by measuring BFR torniquet pressure and the incidence of side effects. Efficacy was determined by measuring body anthropometry and knee isokinetic dynamometry. Feasibility and safety outcomes were reported descriptively or as a proportion with 95% confidence intervals (95% CI), with mean change, 95% CIs, and effect sizes for efficacy outcomes. RESULTS: Twelve participants (mean age 24.8 years [6.5]) were successfully recruited; 11 completed the study. 134/136 sessions were completed (adherence = 98.5%) and 100% of data were collected. There was one event of excessive pain during exercise (0.7%, 95% CI 0.0%, 4.0%), two events of excessive pain post-exercise (1.5%, 95% CI 0.4%, 5.5%), and one event of persistent paraesthesia post-exercise (0.7%, 95% CI 0.0%, 4.0%). Mean maximal BFR torniquet pressure was <200 mmHg. We observed an increase in knee extension peak torque (mean change 12.4 Nm), but no notable changes in body anthropometry. CONCLUSIONS: Lower-limb garment-integrated BFR-training is feasible, has no signal of important harm, and could be used independently.

Preoperative enoxaparin is safe to use in major gynecologic surgery for prophylaxis of venous thromboembolism: a retrospective cohort study.

OBJECTIVE: To evaluate the safety of preoperative enoxaparin in patients undergoing major gynecologic oncology surgery. METHODS: We identified a retrospective cohort group of patients undergoing major gynecologic oncology surgery from June 2002 to June 2004. Exclusion criteria included laparoscopic surgery, inferior vena cava filter, history of venous thromboembolism, and current anticoagulation for prior venous thromboembolism. All patients received prophylaxis with sequential pneumatic compression devices and early ambulation. We identified patients who received (preoperative and postoperative) enoxaparin (20-40 mg) and compared them to patients who received no additional prophylaxis other than pneumatic compression alone. Patient outcomes including estimated blood loss, blood transfusions, operative time, and length of hospital stay were collected. Statistical analysis was performed using the χ Wilcoxon rank sum tests. This study was approved by the institutional review board. RESULTS: We identified 122 patients who met our study criteria; there were 63 patients who received preoperative enoxaparin and 59 patients who received no additional prophylaxis. Both groups were similar in age, body mass index, race, comorbidities, cancer diagnosis, and surgical procedure. There was no significant difference between the enoxaparin group and the sequential pneumatic compression devices-only group regarding transfusion rates (29% and 27%; P = 0.86), operating time (150 and 140 minutes; P = 0.16), blood loss greater than 500 cc (35% and 37%; P = 0.79), and length of stay (5 vs 6 days). CONCLUSION: The use of preoperative enoxaparin is not associated with increased blood loss, transfusion requirements, operative time, or hospital stay among patients having major gynecologic surgery.

Resident sign-out and patient hand-offs: opportunities for improvement.

BACKGROUND: Inpatient care is characterized by multiple transitions of patient care responsibilities. In most residency programs trainees manage transitions via verbal, written, or combined methods of communication termed "sign-out." Often sign-out occurs without standardization or supervision. PURPOSE: The purpose was to assess daily sign-out with a goal of identifying aspects of this process most in need of improvement. METHODS: This was a prospective, observational cohort study of interns' sign-out conducted by industrial engineering students. Daily sign-out was analyzed for inclusion of multiple criteria and scored on organization (on a scale of 0-4) based on how effectively written information was conveyed. RESULTS: We observed 124 unique verbal and written sign-outs. We found that 99% of sign-outs included a general hospital course. Sign-outs were well organized with a mean of 3.1, though substantial variation was noted (SD = 0.8). Directions for anticipated patient events were included in only 42% of sign-outs. Do Not Resuscitate (DNR) or advanced directive discussions were reported in only 11% of sign-outs. Only 50% of successive daily sign-outs were updated. CONCLUSIONS: We found variability in the content and organization of interns' sign-out, possibly reflecting a lack of instruction and supervision. Standardization of sign-out content, and education on good sign-out skills are increasingly important as patient hand-offs become more frequent.

Examining the Public's Most Frequently Asked Questions Regarding COVID-19 Vaccines Using Search Engine Analytics in the United States: Observational Study.

BACKGROUND: The emergency authorization of COVID-19 vaccines has offered the first means of long-term protection against COVID-19-related illness since the pandemic began. It is important for health care professionals to understand commonly held COVID-19 vaccine concerns and to be equipped with quality information that can be used to assist in medical decision-making. OBJECTIVE: Using Google's RankBrain machine learning algorithm, we sought to characterize the content of the most frequently asked questions (FAQs) about COVID-19 vaccines evidenced by internet searches. Secondarily, we sought to examine the information transparency and quality of sources used by Google to answer FAQs on COVID-19 vaccines. METHODS: We searched COVID-19 vaccine terms on Google and used the "People also ask" box to obtain FAQs generated by Google's machine learning algorithms. FAQs are assigned an "answer" source by Google. We extracted FAQs and answer sources related to COVID-19 vaccines. We used the Rothwell Classification of Questions to categorize questions on the basis of content. We classified answer sources as either academic, commercial, government, media outlet, or medical practice. We used the Journal of the American Medical Association's (JAMA's) benchmark criteria to assess information transparency and Brief DISCERN to assess information quality for answer sources. FAQ and answer source type frequencies were calculated. Chi-square tests were used to determine associations between information transparency by source type. One-way analysis of variance was used to assess differences in mean Brief DISCERN scores by source type. RESULTS: Our search yielded 28 unique FAQs about COVID-19 vaccines. Most COVID-19 vaccine-related FAQs were seeking factual information (22/28, 78.6%), specifically about safety and efficacy (9/22, 40.9%). The most common source type was media outlets (12/28, 42.9%), followed by government sources (11/28, 39.3%). Nineteen sources met 3 or more JAMA benchmark criteria with government sources as the majority (10/19, 52.6%). JAMA benchmark criteria performance did not significantly differ among source types (χ 2 4=7.40; P=.12). One-way analysis of variance revealed a significant difference in mean Brief DISCERN scores by source type (F 4,23=10.27; P<.001). CONCLUSIONS: The most frequently asked COVID-19 vaccine-related questions pertained to vaccine safety and efficacy. We found that government sources provided the most transparent and highest-quality web-based COVID-19 vaccine-related information. Recognizing common questions and concerns about COVID-19 vaccines may assist in improving vaccination efforts.

Why does the zebrafish cloche mutant develop lens cataract?

The zebrafish has become a valuable model for examining ocular lens development, physiology and disease. The zebrafish cloche mutant, first described for its loss of hematopoiesis, also shows reduced eye and lens size, interruption in lens cell differentiation and a cataract likely caused by abnormal protein aggregation. To facilitate the use of the cloche mutant for studies on cataract development and prevention we characterized variation in the lens phenotype, quantified changes in gene expression by qRT-PCR and RNA-Seq and compared the ability of two promoters to drive expression of introduced proteins into the cloche lens. We found that the severity of cloche embryo lens cataract varied, while the decrease in lens diameter and retention of nuclei in differentiating lens fiber cells was constant. We found very low expression of both αB-crystallin genes (cryaba and cryabb) at 4 days post fertilization (dpf) by both qRT-PCR and RNA-Seq in cloche, cloche sibling and wildtype embryos and no significant difference in αA-crystallin (cryaa) expression. RNA-Seq analysis of 4 dpf embryos identified transcripts from 25,281 genes, with 1,329 showing statistically significantly different expression between cloche and wildtype samples. Downregulation of eight lens ß- and γM-crystallin genes and 22 retinal related genes may reflect a general reduction in eye development and growth. Six stress response genes were upregulated. We did not find misregulation of any known components of lens development gene regulatory networks. These results suggest that the cloche lens cataract is not caused by loss of αA-crystallin or changes to lens gene regulatory networks. Instead, we propose that the cataract results from general physiological stress related to loss of hematopoiesis. Our finding that the zebrafish αA-crystallin promoter drove strong GFP expression in the cloche lens demonstrates its use as a tool for examining the effects of introduced proteins on lens crystallin aggregation and cataract prevention.

Emergency department discharge prescription errors in an academic medical center.

This study described discharge prescription medication errors written for emergency department patients. This study used content analysis in a cross-sectional design to systematically categorize prescription errors found in a report of 1000 discharge prescriptions submitted in the electronic medical record in February 2015. Two pharmacy team members reviewed the discharge prescription list for errors. Open-ended data were coded by an additional rater for agreement on coding categories. Coding was based upon majority rule. Descriptive statistics were used to address the study objective. Categories evaluated were patient age, provider type, drug class, and type and time of error. The discharge prescription error rate out of 1000 prescriptions was 13.4%, with "incomplete or inadequate prescription" being the most commonly detected error (58.2%). The adult and pediatric error rates were 11.7% and 22.7%, respectively. The antibiotics reviewed had the highest number of errors. The highest within-class error rates were with antianginal medications, antiparasitic medications, antacids, appetite stimulants, and probiotics. Emergency medicine residents wrote the highest percentage of prescriptions (46.7%) and had an error rate of 9.2%. Residents of other specialties wrote 340 prescriptions and had an error rate of 20.9%. Errors occurred most often between 10:00 am and 6:00 pm.

Pre and post hoc analysis of electronic health record implementation on emergency department metrics.

Longitudinal time-based emergency department (ED) performance measures were quantified 12 months before and 12 months after (March 2012-February 2014) implementation of a Meditech 6.0® electronic health record (EHR) at a single urban academic ED. Data assessed were length of stay from door to door, door to admission, door to bed, bed to provider, provider to disposition, and disposition to admission, as well as number of patients leaving against medical advice and number of patients leaving without being seen. Analysis of variance was used to compare levels before and after EHR implementation for each variable, with adjustments made for the number of admissions, transfers, and month. No difference was seen in monthly volume, admissions, or transfers. Implementation of an EHR resulted in a sustained increase in ED time metrics for mean length of stay and times from door to door, door to admission, door to bed, and provider to disposition. Decreased ED time metrics were seen in bed-to-provider and disposition-to-admit times. The number of patients who left against medical advice increased after implementation, but the number of patients who left without being seen was not significantly different. Thus, EHR implementation was associated with an increase in time with most performance metrics. Although general times trended back to near preimplementation baselines, most ED time metrics remained elevated beyond the study length of 12 months. Understanding the impact of EHR system implementation on the overall performance of an ED can help departments prepare for potential adverse effects of such systems on overall efficiency.

Methodological quality of systematic reviews referenced in clinical practice guidelines for the treatment of opioid use disorder.

INTRODUCTION: With efforts to combat opioid use disorder, there is an increased interest in clinical practice guidelines (CPGs) for opioid use disorder treatments. No literature exists examining the quality of systematic reviews used in opioid use disorder CPGs. This study aims to describe the methodological quality and reporting clarity of systematic reviews (SRs) used to create CPGs for opioid use disorder. METHODS: From June to July 2016 guideline clearinghouses and medical literature databases were searched for relevant CPGs used in the treatment of opioid use disorder. Included CPGs must have been recognized by a national organization. SRs from the reference section of each CPG was scored by using AMSTAR (a measurement tool to assess the methodological quality of systematic reviews) tool and PRISMA (preferred reporting items for systematic reviews and meta-analyses) checklist. RESULTS: Seventeen CPGs from 2006-2016 were included in the review. From these, 57 unique SRs were extracted. SRS comprised 0.28% to 17.92% of all references found in the CPGs. All SRs obtained moderate or high methodological quality score on the AMSTAR tool. All reviews met at least 70% of PRISMA criteria. In PRISMA, underperforming areas included accurate title labeling, protocol registration, and risk of bias. Underperforming areas in AMSTAR included conflicts of interest, funding, and publication bias. A positive correlation was found between AMSTAR and PRISMA scores (r = .79). CONCLUSION: Although the SRs in the CPGs were of good quality, there are still areas for improvement. Systematic reviewers should consult PRISMA and AMSTAR when conducting and reporting reviews. It is important for CPG developers to consider methodological quality as a factor when developing CPG recommendations, recognizing that the quality of systematic reviews underpinning guidelines does not necessarily correspond to the quality of the guideline itself.

The zebrafish as a model system for analyzing mammalian and native α-crystallin promoter function.

Previous studies have used the zebrafish to investigate the biology of lens crystallin proteins and their roles in development and disease. However, little is known about zebrafish α-crystallin promoter function, how it compares to that of mammals, or whether mammalian α-crystallin promoter activity can be assessed using zebrafish embryos. We injected a variety of α-crystallin promoter fragments from each species combined with the coding sequence for green fluorescent protein (GFP) into zebrafish zygotes to determine the resulting spatiotemporal expression patterns in the developing embryo. We also measured mRNA levels and protein abundance for all three zebrafish α-crystallins. Our data showed that mouse and zebrafish αA-crystallin promoters generated similar GFP expression in the lens, but with earlier onset when using mouse promoters. Expression was also found in notochord and skeletal muscle in a smaller percentage of embryos. Mouse αB-crystallin promoter fragments drove GFP expression primarily in zebrafish skeletal muscle, with less common expression in notochord, lens, heart and in extraocular regions of the eye. A short fragment containing only a lens-specific enhancer region increased lens and notochord GFP expression while decreasing muscle expression, suggesting that the influence of mouse promoter control regions carries over into zebrafish embryos. The two paralogous zebrafish αB-crystallin promoters produced subtly different expression profiles, with the aBa promoter driving expression equally in notochord and skeletal muscle while the αBb promoter resulted primarily in skeletal muscle expression. Messenger RNA for zebrafish αA increased between 1 and 2 days post fertilization (dpf), αBa increased between 4 and 5 dpf, but αBb remained at baseline levels through 5 dpf. Parallel reaction monitoring (PRM) mass spectrometry was used to detect αA, aBa, and αBb peptides in digests of zebrafish embryos. In whole embryos, αA-crystallin was first detected by 2 dpf, peaked in abundance by 4-5 dpf, and was localized to the eye. αBa was detected in whole embryo at nearly constant levels from 1-6 dpf, was also localized primarily to the eye, and its abundance in extraocular tissues decreased from 4-7 dpf. In contrast, due to its low abundance, no αBb protein could be detected in whole embryo, or dissected eye and extraocular tissues. Our results show that mammalian α-crystallin promoters can be efficiently screened in zebrafish embryos and that their controlling regions are well conserved. An ontogenetic shift in zebrafish aBa-crystallin promoter activity provides an interesting system for examining the evolution and control of tissue specificity. Future studies that combine these promoter based approaches with the expanding ability to engineer the zebrafish genome via techniques such as CRISPR/Cas9 will allow the manipulation of protein expression to test hypotheses about lens crystallin function and its relation to lens biology and disease.

Recruiting and retaining participants in a combined behavioral and pharmacological clinical trial.

OBJECTIVE: This article focuses on the practical and methodological considerations that went into recruiting and retaining participants in a complex clinical trial. It describes multiple recruitment strategies that were employed to meet stringent eligibility criteria and manage efficiently the flow of participants to fulfill recruitment goals. Both study-wide and site-specific methods that were employed to maximize retention rates are discussed. Also discussed is how the coordinating center worked together with study investigators to improve recruitment and retention. METHOD: A multidimensional perspective on recruitment and retention is discussed. Details are provided on how barriers to recruitment and retention were addressed. Intervention strategies designed by the coordinating center to meet performance goals are described. Data are presented to support the utility of recruitment and retention methods. The COMBINE Study served as a case illustration for various issues addressed in the article. RESULTS: As a result of the combined efforts of the coordinating center and study sites, there was steady improvement in recruitment and retention rates over the course of the trial. The recruitment goal of 1,375 participants was surpassed by September 2003. Also, improvement in data completeness rates improved by 18-27 percentage points over a 3-year period, culminating in follow-up rates meeting or exceeding performance goals by 2004. CONCLUSIONS: Various strategies and procedures designed to address recruitment and retention problems in large, complex combination pharmacotherapy and behavioral trials were effective in meeting recruitment and retention goals. The lessons learned may be of value to investigators not only involved in multisite, combination trials but in monotherapy and/or single-site studies as well.

Developing a baseline assessment battery: balancing patient time burden with essential clinical and research monitoring.

OBJECTIVE: Baseline assessment in a multisite alcohol-dependence treatment study has several purposes: addressing inclusion/exclusion criteria and characterizing participants to illuminate subsequent efficacy and safety patterns of the interventions. Combination pharmacotherapy and behavioral therapy trials, however, require more complex assessments than single-modality studies. Medication trials require measures of initial safety for study drug as well as for subsequent side-effect and adverse-effect monitoring (e.g., physical examination, laboratory markers, somatic symptoms, medical conditions and concomitant medications). Behavioral therapy trials (and in some cases medication trials) warrant baseline measurement of mediators of therapy effect (e.g., prior treatment history, motivation for change, self-efficacy, other psychiatric conditions, treatment expectations and network supports). Measures may be needed to interpret interactions that may be discovered between these modalities. METHOD: The National Institute on Alcohol Abuse and Alcoholism COMBINE Study evaluated the potentially overwhelming number of candidate instruments through an iterative process, using the following sequence to finalize a rational baseline assessment battery: key constructs, representative measures, determination of duration of assessment, risk of assessment reactivity, goal priorities, necessity for measure "pruning" and order of measure presentation. After selecting the draft battery, feasibility was evaluated in a pilot study prior to the main trial. RESULTS: The battery was feasible to administer and avoided unintended selection bias. Dropout was substantial, however, and differences across sites in baseline assessment completion rates reflected a tendency of a central intake model to function as an initial filter of dropouts, compared with a direct recruitment model. CONCLUSIONS: This process holds several potentially useful lessons for investigators.