Distinct p53 transcriptional programs dictate acute DNA-damage responses and tumor suppression.

The molecular basis for p53-mediated tumor suppression remains unclear. Here, to elucidate mechanisms of p53 tumor suppression, we use knockin mice expressing an allelic series of p53 transcriptional activation mutants. Microarray analysis reveals that one mutant, p53(25,26), is severely compromised for transactivation of most p53 target genes, and, moreover, p53(25,26) cannot induce G(1)-arrest or apoptosis in response to acute DNA damage. Surprisingly, p53(25,26) retains robust activity in senescence and tumor suppression, indicating that efficient transactivation of the majority of known p53 targets is dispensable for these pathways. In contrast, the transactivation-dead p53(25,26,53,54) mutant cannot induce senescence or inhibit tumorigenesis, like p53 nullizygosity. Thus, p53 transactivation is essential for tumor suppression but, intriguingly, in association with a small set of novel p53 target genes. Together, our studies distinguish the p53 transcriptional programs involved in acute DNA-damage responses and tumor suppression-a critical goal for designing therapeutics that block p53-dependent side effects of chemotherapy without compromising p53 tumor suppression.

Global genomic profiling reveals an extensive p53-regulated autophagy program contributing to key p53 responses.

The mechanisms by which the p53 tumor suppressor acts remain incompletely understood. To gain new insights into p53 biology, we used high-throughput sequencing to analyze global p53 transcriptional networks in primary mouse embryo fibroblasts in response to DNA damage. Chromatin immunoprecipitation sequencing reveals 4785 p53-bound sites in the genome located near 3193 genes involved in diverse biological processes. RNA sequencing analysis shows that only a subset of p53-bound genes is transcriptionally regulated, yielding a list of 432 p53-bound and regulated genes. Interestingly, we identify a host of autophagy genes as direct p53 target genes. While the autophagy program is regulated predominantly by p53, the p53 family members p63 and p73 contribute to activation of this autophagy gene network. Induction of autophagy genes in response to p53 activation is associated with enhanced autophagy in diverse settings and depends on p53 transcriptional activity. While p53-induced autophagy does not affect cell cycle arrest in response to DNA damage, it is important for both robust p53-dependent apoptosis triggered by DNA damage and transformation suppression by p53. Together, our data highlight an intimate connection between p53 and autophagy through a vast transcriptional network and indicate that autophagy contributes to p53-dependent apoptosis and cancer suppression.

Inappropriate p53 activation during development induces features of CHARGE syndrome.

CHARGE syndrome is a multiple anomaly disorder in which patients present with a variety of phenotypes, including ocular coloboma, heart defects, choanal atresia, retarded growth and development, genitourinary hypoplasia and ear abnormalities. Despite 70-90% of CHARGE syndrome cases resulting from mutations in the gene CHD7, which encodes an ATP-dependent chromatin remodeller, the pathways underlying the diverse phenotypes remain poorly understood. Surprisingly, our studies of a knock-in mutant mouse strain that expresses a stabilized and transcriptionally dead variant of the tumour-suppressor protein p53 (p53(25,26,53,54)), along with a wild-type allele of p53 (also known as Trp53), revealed late-gestational embryonic lethality associated with a host of phenotypes that are characteristic of CHARGE syndrome, including coloboma, inner and outer ear malformations, heart outflow tract defects and craniofacial defects. We found that the p53(25,26,53,54) mutant protein stabilized and hyperactivated wild-type p53, which then inappropriately induced its target genes and triggered cell-cycle arrest or apoptosis during development. Importantly, these phenotypes were only observed with a wild-type p53 allele, as p53(25,26,53,54)(/-) embryos were fully viable. Furthermore, we found that CHD7 can bind to the p53 promoter, thereby negatively regulating p53 expression, and that CHD7 loss in mouse neural crest cells or samples from patients with CHARGE syndrome results in p53 activation. Strikingly, we found that p53 heterozygosity partially rescued the phenotypes in Chd7-null mouse embryos, demonstrating that p53 contributes to the phenotypes that result from CHD7 loss. Thus, inappropriate p53 activation during development can promote CHARGE phenotypes, supporting the idea that p53 has a critical role in developmental syndromes and providing important insight into the mechanisms underlying CHARGE syndrome.

Fast and accurate HLA typing from short-read next-generation sequence data with xHLA.

The HLA gene complex on human chromosome 6 is one of the most polymorphic regions in the human genome and contributes in large part to the diversity of the immune system. Accurate typing of HLA genes with short-read sequencing data has historically been difficult due to the sequence similarity between the polymorphic alleles. Here, we introduce an algorithm, xHLA, that iteratively refines the mapping results at the amino acid level to achieve 99-100% four-digit typing accuracy for both class I and II HLA genes, taking only [Formula: see text]3 min to process a 30× whole-genome BAM file on a desktop computer.

Intermittent Versus Continuous and Intermittent Medications for Pain and Sedation After Pediatric Cardiothoracic Surgery; A Randomized Controlled Trial.

OBJECTIVES: Compare continuous infusions of morphine and midazolam in addition to intermittent doses with an intermittent only strategy for pain and sedation after pediatric cardiac surgery. DESIGN: Randomized controlled trial. SETTING: Advocate Children's Hospital, Oak Lawn, IL. PATIENTS: Sixty patients 3 months to 4 years old with early extubation after pediatric cardiac surgery. INTERVENTIONS: Patients received a continuous infusion of morphine and midazolam or placebo for 24 hours. Both groups received intermittent morphine and midazolam doses as needed. MEASUREMENTS AND MAIN RESULTS: Gender, age, bypass time, and surgical complexity were not different between groups. Scheduled ketorolac and acetaminophen were used in both groups and were not associated with adverse events. The mean, median, and maximum Faces, Legs, Activity, Cry, And Consolability score were not different between groups. There was no significant difference in number of intermittent doses received between groups. The total morphine dose was higher in the continuous/intermittent group (0.90 vs 0.23 mg/kg; p < 0.01). The total midazolam dose was also higher in the continuous/intermittent group (0.90 vs 0.18 mg/kg; p < 0.01). The hospital length of stay was longer in the continuous/intermittent group (8.4 vs 4.9 d; p = 0.04). CONCLUSIONS: Pain was not better controlled with the addition of continuous infusions of morphine and midazolam when compared with intermittent dosing only. Use of continuous infusions resulted in a significantly higher total dosage of these medications and a longer length of stay.

The pedagogical value of testing: how far does it extend?

Information is generally more memorable after it is studied and tested than when it is only studied. One must be cautious to use this phenomenon strategically, however, due to uncertainty about whether testing improves memorability for only tested material, facilitates learning of related non-tested content, or inhibits memory of non-tested material. 52 second-year Pharmacy students were asked to study therapeutic aspects of gastroesophageal reflux disease and peptic ulcer disease. One group was given 30 min to study. Another was given 20 min to study and 10 min to complete a 10-item test. Two weeks later a 40-item test was delivered to both groups that contained (a) the 10 learning phase questions, (b) 10 new questions drawn from the studied material, (c) 10 new questions about therapeutics in different disease states, and (d) 10 new questions drawn from more general pharmaceutical knowledge (e.g., basic physiology and drug characteristics). Moderate to large retrieval-enhanced learning effects were observed for both questions about material that was tested (22.9% difference in scores, p < 0.05, d = 0.60) and questions about material that was studied without being tested (18.9% difference, p < 0.05, d = 0.75). Such effects were not observed for questions that were not part of the study material: therapeutic questions that addressed different disease states (1.8% difference, p > 0.7, d = 0.08) or generic pharmaceutical questions (7.4% difference, p > 0.2, d = 0.32). Being tested made it more likely that students would report reviewing the material after the initial learning session, but such reports were not associated with better test performance. The benefit of mentally retrieving information from studied material appears to facilitate the retrieval of information that was studied without being tested. Such generalization of the benefit of testing can increase the flexibility of test-based pedagogic interventions.

How transplant centers deal with the dextran shortage: recommendations for comparing alternatives.

BACKGROUND: In the United States, dextran 40 in 0.9% NaCl is the preferred reagent for the thawing and preparation of cord blood units for hematopoietic stem cell transplantation. The recurring nationwide shortage of this reagent could have implications that extend to the avoidance of cord blood for transplantation. STUDY DESIGN AND METHODS: To address the shortage, the National Marrow Donor Program and its Cord Blood Advisory Group sought to identify available alternative reagents or manufacturers. A sample of transplant centers (TCs) were surveyed to determine their process to compare these alternatives. The TCs were then asked to share their comparability protocols for review. RESULTS: The 12 TCs that responded to the survey studied various types of alternative reagents and manufacturers of the standard dextran 40 in 0.9% NaCl. Four TCs submitted their protocols from which a model comparability protocol was created for centers who need assistance. CONCLUSION: Whether comparing dextran 40 in 0.9% NaCl to that of a different manufacturer or a different reagent, the results of the comparability studies submitted by the TCs indicated equivalency. During a shortage, the model comparability study protocol can be used as a reference to establish an alternative to dextran 40 in 0.9% NaCl.

Full p53 transcriptional activation potential is dispensable for tumor suppression in diverse lineages.

Over half of all human cancers, of a wide variety of types, sustain mutations in the p53 tumor suppressor gene. Although p53 limits tumorigenesis through the induction of apoptosis or cell cycle arrest, its molecular mechanism of action in tumor suppression has been elusive. The best-characterized p53 activity in vitro is as a transcriptional activator, but the identification of numerous additional p53 biochemical activities in vitro has made it unclear which mechanism accounts for tumor suppression. Here, we assess the importance of transcriptional activation for p53 tumor suppression function in vivo in several tissues, using a knock-in mouse strain expressing a p53 mutant compromised for transcriptional activation, p53(25,26). p53(25,26) is severely impaired for the transactivation of numerous classical p53 target genes, including p21, Noxa, and Puma, but it retains the ability to activate a small subset of p53 target genes, including Bax. Surprisingly, p53(25,26) can nonetheless suppress tumor growth in cancers derived from the epithelial, mesenchymal, central nervous system, and lymphoid lineages. Therefore, full transactivation of most p53 target genes is dispensable for p53 tumor suppressor function in a range of tissue types. In contrast, a transcriptional activation mutant that is completely defective for transactivation, p53(25,26,53,54), fails to suppress tumor development. These findings demonstrate that transcriptional activation is indeed broadly critical for p53 tumor suppressor function, although this requirement reflects the limited transcriptional activity observed with p53(25,26) rather than robust transactivation of a full complement of p53 target genes.

Benchmarking Enrichment Efforts in the US & Canada Across Species and Enrichment Categories.

Enrichment is important for animal welfare and data quality. Provision of enrichment opportunities varies between species and enrichment category. However, data benchmarking these differences does not exist. Our objective was to characterize enrichment provision and associated factors across species in the US and Canada. Personnel who work with research animals (n = 1098) in the US and Canada voluntarily responded to online promotions and completed a survey about enrichment used for the species they worked with most, their control of and wish for more enrichment, stress or pain in the animals they worked the most with, and demographics. All participants (except those working with rats) received the same questionnaire regardless of species to allow objectivity, as the effects of many enrichment items on some species have not yet been determined. The questionnaire asked about enrichments that were beneficial to at least one species. The provision of enrichment was allocated into 2 outcome variables: diversity and frequency per enrichment category. Results showed a significant interaction between enrichment category and species. Generally, physical, nutritional, and sensory enrichments were provided less often than social enrichment. In addition, nonhuman primates received more diverse and more frequent enrichment than did other species (twice as much as rats and mice). Enrichment was provided less frequently by personnel who wished they could do more than the status quo. Both enrichment frequency and diversity were higher in respondents from Canada, those who had more control over provision, and those who had been in the field longer. While our results cannot be used to determine the quality of enrichment provided to various species, they do provide information on current enrichment practices in the US and Canada and identify differences in implementation by species and enrichment category. The data also indicate provision of enrichment is influenced by factors such as country and individual control over enrichment. This information can also be used to identify areas for greater enrichment efforts for some species (for example, rats and mice) and categories, with the ultimate goal of improving animal welfare.

Effect of HLA-matching recipients to donor noninherited maternal antigens on outcomes after mismatched umbilical cord blood transplantation for hematologic malignancy.

Transplantation-related mortality (TRM) is high after HLA-mismatched umbilical cord blood (UCB) transplantation (UCBT). In utero, exposure to noninherited maternal antigen (NIMA) is recognized by the fetus, which induces T regulator cells to that haplotype. It is plausible that UCBTs in which recipients are matched to donor NIMAs may alleviate some of the excess mortality associated with this treatment. To explore this concept, we used marginal matched-pair Cox regression analysis to compare outcomes in 48 NIMA-matched UCBTs (ie, the NIMA of the donor UCB unit matched to the patient) and in 116 non-NIMA-matched UCBTs. All patients had a hematologic malignancy and received a single UCB unit. Cases and controls were matched on age, disease, disease status, transplantation-conditioning regimen, HLA match, and infused cell dose. TRM was lower after NIMA-matched UCBTs compared with NIMA-mismatched UCBTs (relative risk, 0.48; P = .05; 18% versus 32% at 5 years posttransplantation). Consequently, overall survival was higher after NIMA-matched UCBT. The 5-year probability of overall survival was 55% after NIMA-matched UCBTs versus 38% after NIMA-mismatched UCBTs (P = .04). When faced with the choice of multiple HLA-mismatched UCB units containing adequate cell doses, selecting an NIMA-matched UCB unit may improve survival after mismatched UCBT.

Exploring Perceptions of Equine Welfare Scenarios Using a Positive Approach.

Horse welfare is a sensitive topic that often results in a variety of strong feelings when discussed in the horse-owning public. This study used a scenario-based questionnaire in a positive psychology approach to assess the public's feelings and discussions about horse welfare. Results indicated themes in important welfare qualities such as turnout, shelter, and ability to express natural behaviors, as well as a positive discussion about welfare. This study provides future implications for further research techniques in this area as well as communicative strategies surrounding equine welfare practices.

Guidelines for the development and validation of new potency assays for the evaluation of umbilical cord blood.

The following commentary was developed by the National Marrow Donor Program Cord Blood Advisory Group and is intended to provide an overview of umbilical cord blood (UCB) processing, summarize the current state of potency assays used to characterize UCB, and define limitations of the assays and future needs of the cord blood banking and transplant community. The UCB banking industry is eager to participate in the development of standardized assays to uniformly characterize cellular therapy products that are manufactured in a variety of ways. This paper describes the desired qualities of these assays and how the industry proposes to co-operate with developers to bring relevant assays to market. To that end, the National Marrow Donor Program (NMDP) Cord Blood Bank Network is available to serve as a resource for UCB testing material, research and development consulting, and product/assay testing in an accredited UCB manufacturing environment.

Integrated STEM and poultry science curriculum to increase agricultural literacy.

The shortage of graduates pursuing careers in the poultry industry is linked to a low awareness and lack of interest. Increasing agricultural literacy could promote engagement in future poultry science opportunities. We developed an integrated STEM curriculum within a poultry science context to assess the program's impact on students' agricultural literacy development. The Elementary Education Gain Grow (E.G.G.) program consists of 5 online modules, an interactive notebook, a simulation game, and a team project. In fall 2019, 480 Indiana 4th and 5th grade students enrolled in the pilot program. A 14-point poultry content-based questionnaire was administered online to students prior to program engagement, between online and team project activities, and at program completion. Student content scores (n = 111; 23.13% response rate) increased from 7.99 (SD = 1.85) preprogram to 9.76 (SD = 2.44) post online modules (P < 0.05; Cohen's d = 0.82) and remained constant throughout the remainder of the program. Student notebook responses (n = 172; 35.83% response rate) provided qualitative data of their self-reported agricultural literacy gains and revealed patterns of increased agricultural literacy relating to the program's learning objectives. These results support the program's ability to increase student agricultural literacy. Teacher feedback (n = 9; 69.2% response rate) suggests that teachers agreed with the program's effectiveness, with qualitative responses highlighting individual experiences. Our pilot program findings support the use of an integrated STEM and poultry science elementary curriculum to increase student agricultural literacy as well as demonstrate the effectiveness of the program as an educational resource.

Changing Human Behavior to Improve Animal Welfare: A Longitudinal Investigation of Training Laboratory Animal Personnel about Heterospecific Play or "Rat Tickling".

Despite evidence for rat tickling's animal welfare benefits, the technique is rarely implemented in part because of a lack of training. This study's purpose was to determine the efficacy of online-only or online + hands-on training programs on key outcomes for rat tickling in comparison to a waitlist control condition. After completing a baseline survey, laboratory animal personnel currently working with rats in the United States were semi-randomized to receive online-only training (n = 30), online + hands-on training (n = 34), or waitlist control (n = 32). Participants received further surveys directly after training and 2 months later. Data were analyzed using general linear mixed models. At the 2-month follow-up compared to baseline, both training groups reported increased implementation, self-efficacy, knowledge, and familiarity of rat tickling while only the online + hands-on training participants reported increased control beliefs (while the waitlist group stayed the same). At the 2-month follow-up compared to the waitlist, hands-on training participants reported increased self-efficacy and familiarity with rat tickling. Overall, findings show that both online-only and online + hands-on training can improve key outcomes for rat tickling. Although online + hands-on training is slightly more effective, the interactive online-only training has the potential to improve widescale implementation of a welfare-enhancing technique.

Laboratory Animal Welfare Meets Human Welfare: A Cross-Sectional Study of Professional Quality of Life, Including Compassion Fatigue in Laboratory Animal Personnel.

Laboratory animal personnel may experience significant stress from working with animals in scientific research. Workplace stress can be assessed by evaluating professional quality of life, which is comprised of compassion fatigue (i.e., burnout and secondary traumatic stress) and compassion satisfaction. This research aimed to explore the associations between risk factors and professional quality of life in laboratory animal personnel. In a cross-sectional, convenience sample design, laboratory animal personnel were recruited from widespread online promotion. A total of 801 personnel in the United States or Canada completed an online survey regarding professional quality of life, social support, euthanasia, enrichment, stress/pain levels, and human-animal interactions. Participants worked in a wide range of settings (e.g., industry, academia), research types (e.g., basic, applied, regulatory), species (e.g., non-human primates, mice), and roles (e.g., animal caretaker, veterinarian). Data were analyzed using general linear models. Personnel who reported higher compassion fatigue also reported lower social support, higher animal stress/pain, higher desire to implement more enrichment, and less control over performing euthanasia (p's < 0.05). Higher burnout was associated with less diverse/frequent enrichment, using physical euthanasia methods, and longer working hours. Higher secondary traumatic stress was associated with more relationship-promoting human-animal interactions (e.g., naming animals) and working as a trainers (p's < 0.05). Higher compassion satisfaction was associated with higher social support, less animal stress/pain, and more human-animal interactions (p's < 0.05). Surprisingly, neither personnel's primary animal type (e.g., non-human primates, mice) nor frequency of euthanasia (e.g., daily, monthly) were associated with professional quality of life (p's > 0.05). Our findings show that the professional quality of life of laboratory animal personnel is associated with several factors. Personnel reporting poorer professional quality of life also reported less social support, higher animal stress/pain, less enrichment diversity/frequency and wished they could provide more enrichment, using physical euthanasia, and less control over performing euthanasia. Poorer professional quality of life was also seen in personnel working as trainers, at universities, and longer hours. This study contributes important empirical data that may provide guidance for developing interventions (e.g., improved social support, decreased animal stress, increased animal enrichment diversity/frequency, greater control over euthanasia) to improve laboratory animal personnel's professional quality of life.

Laboratory animal welfare and human attitudes: A cross-sectional survey on heterospecific play or "rat tickling".

INTRODUCTION: Laboratory rat welfare is critically influenced by laboratory animal personnel through their implementation, or lack of implementation, of various enrichment techniques. One such promising technique is heterospecific play, or "rat tickling", which mimics aspects of rat rough-and-tumble play and can contribute to improving welfare, but may be infrequently implemented. The theory of planned behavior can be used to study implementation by measuring intentions and beliefs about rat tickling, including behavioral attitudes (whether it is good or bad), subjective norms (whether there is social/professional pressure to provide it), and control beliefs (whether they feel in control of providing it). Therefore, the objective of this study was to identify current rat tickling prevalence and predictors among laboratory animal personnel in the United States and Canada. Our hypothesis was that rat tickling prevalence would be low and associated with beliefs about the practice, enrichment, and laboratory animals in general. METHODS: Laboratory animal personnel were recruited from widespread online promotion. A total of 794 personnel (mean = 40±11 years, 80% white, 80% female) completed at least 50% of the mixed methods online survey and met inclusion criteria of currently working with laboratory rats in the USA or Canada. The survey included questions about demographics, enrichment practices and beliefs, attitudes towards rats, general positive behaviors (e.g. talking to laboratory animals), and both practices and beliefs about rat tickling. Qualitative data were coded using thematic analysis. Quantitative data were analyzed using general linear models. RESULTS: Laboratory personnel reported low levels of rat tickling implementation, with 89% of participants reporting using it never or rarely. Laboratory personnel reported 2 key benefits (handling: 61%, welfare: 55%) and 3 key barriers (time: 59%, personnel: 22%, and research: 22%) to rat tickling using qualitative analysis. Current and planned rat tickling were positively associated with more positive beliefs (social/professional pressure p<0.0001, control of providing tickling p<0.0001) and familiarity with tickling (p<0.0001). Current rat tickling was also positively associated with more positive general behaviors towards laboratory animals, such as naming animals (p<0.0001). Future rat tickling was positively associated with more positive attitudes about it (p<0.0001) and a desire to implement more enrichment (p<0.01). CONCLUSION: Our findings show that even though rat tickling implementation is currently low, it is positively associated with personnel beliefs, familiarity, general attitudes, and a desire for more enrichment. That is, laboratory animal personnel were more likely to provide rat tickling if they were more familiar with it, thought providing it was both good and under their control, and felt subject to social/professional pressure, as well as if they wanted to provide more enrichment and generally had more positive behaviors towards laboratory animals. There is potential to increase rat tickling by increasing personnel familiarity with the procedure through training, decreasing the time required, and changing personnel beliefs-thereby improving rat welfare.

Allele-level HLA matching for umbilical cord blood transplantation for non-malignant diseases in children: a retrospective analysis.

BACKGROUND: The standard for selecting unrelated umbilical cord blood units for transplantation for non-malignant diseases relies on antigen-level (lower resolution) HLA typing for HLA-A and HLA-B, and allele-level for HLA-DRB1. We aimed to study the effects of allele-level matching at a higher resolution-HLA-A, HLA-B, HLA-C, and HLA-DRB1, which is the standard used for adult unrelated volunteer donor transplantation for non-malignant diseases-for umbilical cord blood transplantation. METHODS: We retrospectively studied 1199 paediatric donor-recipient pairs with allele-level HLA matching who received a single unit umbilical cord blood transplantation for non-malignant diseases reported to the Center for International Blood and Marrow Transplant Research or Eurocord and European Group for Blood and Marrow Transplant. Transplantations occurred between Jan 1, 2000, and Dec 31, 2012. The primary outcome was overall survival. The effect of HLA matching on survival was studied using a Cox regression model. FINDINGS: Compared with HLA-matched transplantations, mortality was higher with transplantations mismatched at two (hazard ratio [HR] 1·55, 95% CI 1·08-2·21, p=0·018), three (2·04, 1·44-2·89, p=0·0001), and four or more alleles (3·15, 2·16-4·58, p<0·0001). There were no significant differences in mortality between transplantations that were matched and mismatched at one allele (HR 1·18, 95% CI 0·80-1·72, p=0·39). Other factors associated with higher mortality included recipient cytomegalovirus seropositivity (HR 1·40, 95% CI 1·13-1·74, p=0·0020), reduced intensity compared with myeloablative conditioning regimens (HR 1·36, 1·10-1·68, p=0·0041), transplantation of units with total nucleated cell dose of more than 21 × 107 cells per kg compared with 21 × 107 cells per kg or less (HR 1·47, 1·11-1·95, p=0·0076), and transplantations done in 2000-05 compared with those done in 2006-12 (HR 1·64, 1·31-2·04, p<0·0001). The 5-year overall survival adjusted for recipient cytomegalovirus serostatus, conditioning regimen intensity, total nucleated cell dose, and transplantation period was 79% (95% CI 74-85) after HLA matched, 76% (71-81) after one allele mismatched, 70% (65-75) after two alleles mismatched, 62% (57-68) after three alleles mismatched, and 49% (41-57) after four or more alleles mismatched transplantations. Graft failure was the predominant cause of mortality. INTERPRETATION: These data support a change from current practice in that selection of unrelated umbilical cord blood units for transplantation for non-malignant diseases should consider allele-level HLA matching at HLA-A, HLA-B, HLA-C, and HLA-DRB1. FUNDING: National Cancer Institute; National Heart, Lung, and Blood Institute; National Institute for Allergy and Infectious Diseases; US Department of Health and Human Services-Health Resources and Services Administration; and US Department of Navy.

Chemical Screening in Zebrafish.

Phenotypic small molecule screens in zebrafish have gained popularity as an unbiased approach to probe biological processes. In this chapter we outline basic methods for performing chemical screens with larval zebrafish including breeding large numbers of embryos, plating larval fish into multi-well dishes, and adding small molecules to these wells. We also highlight important considerations when designing and interpreting the results of a phenotypic screen and possible follow-up approaches, including popular methods used to identify the mechanism of action of a chemical compound.

Show Horse Welfare: The Viewpoints of Judges, Stewards, and Show Managers.

The purpose of this study was to gain a better understanding of the current state of stock-type show horse welfare based on the perceptions of show officials and to identify potential means of preventing and intervening in compromises to show horse welfare. Thirteen horse show officials, including judges, stewards, and show managers, were interviewed. Findings revealed the officials had an incomplete understanding of nonhuman animal welfare and a high level of concern regarding the public's perception of show horse welfare. The officials attributed most of the frequently observed compromises to show horse welfare to (a) novices', amateurs', and young trainers' lack of experience or expertise, and (b) trainers' and owners' unrealistic expectations and prioritization of winning over horse welfare. The officials emphasized a need for distribution of responsibility among associations, officials, and individuals within the industry. Although the officials noted recent observable positive changes in the industry, they emphasized the need for continued improvements in equine welfare and greater educational opportunities for stakeholders.