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ABSTRACT: The objective of this study was to evaluate the effect of a novel consensus bacterial 6-phytase variant (PhyG) on egg productivity, eggshell quality, and body composition of laying hens fed inorganic phosphate-free diets with reduced energy and nutrients from 23 to 72 wk of age. Five treatments were randomly assigned, performing 28 replicates per treatment with 4 hens each, totaling 560 Hy-Line W80 birds. A positive control (PC) feed was formulated to contain adequate levels of energy and nutrients. A negative control (NC) feed was formulated without added inorganic phosphate (0.12% nonphytic phosphorus [nPP]) and reduced in Ca, Na, dig AA, and metabolizable energy in comparison with PC feed. Phytase was supplemented in the NC feed at 0, 300, 600, and 900 FTU/kg of feed. The responses evaluated were performance, egg quality, economic analysis, body composition, and tibia composition. Data were analyzed by a 2-factor (diet and age) repeated measure analysis. Overall, the feed intake, hen-day egg production, egg mass, and egg revenue were reduced by the complete removal of dicalcium phosphate (DCP) (P < 0.05). Supplement phytase in the NC diet elicits a positive response on each one of those variables. Laying hens consuming the NC feed with 900 FTU/kg of phytase produced more eggs per hen-housed compared with the phytase dosages of 300 and 600 FTU/kg. Body composition was not affected by dietary nPP, Ca, Na, dig AA, and energy reductions (P > 0.05). At 72-wk-old, tibia ash was reduced in hens consuming the NC diet vs. PC (P < 0.05) and no difference was observed between hens supplemented with phytase and the PC feed. Margin over feeding cost increased in a dose-dependent manner with phytase supplementation. Supplementation with 900 FTU/kg of phytase is recommended to improve the number of eggs produced per hen-housed and the number of marketable eggs produced through 23 to 72 wk of age, under this dietary setting.

BackgroundClinical utility of routinely measured serial biomarkers in predicting escalation of inpatient care intensity and mortality among hospitalized patients with COVID-19 remains unknown.MethodsThis retrospective cohort study included patients with COVID-19 who admitted to the Massachusetts General Hospital between March and June 2020 and January to March 2021. White blood cell (WBC) count, platelet count, C-reactive protein (CRP), and D-dimer values were measured on days 1, 3, and 7 of admission. Clinical outcomes include 30- and 60-day morality, ICU transfer, and overall survival (OS) over a follow-up period of 90 days. The association between serial biomarkers and outcomes were assessed using multivariable logistic regression and Cox proportional hazards models.Measurements and main resultsOf the 456 patients hospitalized with COVID-19, 199 (43.6%) were ICU, 179 (39.3%) were medical floor, and 78 (17.1%) were initially admitted to the medical floor and then transferred to the ICU. In adjusted analyses, each unit increase in the slope of CRP was associated with a 42% higher odds of ICU transfer after controlling for the initial admission level (OR = 1.42, 95% CI: 1.25-1.65, P < 0.001). Including serial change in CRP levels from initial level on admission achieved the greatest predictive accuracy for ICU transfer (AUC = 0.72, 95% CI: 0.64-0.79).ConclusionsSerial change in CRP levels from admission is associated with escalations of inpatient care intensity and mortality among hospitalized patients with COVID-19.

Integrated approaches to testing and assessments (IATAs) have been proposed as a method to organise new approach methodologies in order to replace traditional animal testing for chemical safety assessments. To capture the mechanistic aspects of toxicity assessments, IATAs can be framed around the adverse outcome pathway (AOP) concept. To utilise AOPs fully in this context, a sufficient number of pathways need to be present to develop fit for purpose IATAs. In silico approaches can support IATA through the provision of predictive models and also through data integration to derive conclusions using a weight-of-evidence approach. To examine the maturity of a developmental and reproductive toxicity (DART) AOP network derived from the literature, an assessment of its coverage was performed against a novel toxicity dataset. A dataset of diverse compounds, with data from studies performed according to OECD test guidelines TG-421 and TG-422, was curated to test the performance of an in silico model based on the AOP network – allowing for the identification of knowledge gaps within the network. One such gap in the knowledge was filled through the development of an AOP stemming from the molecular initiating event ‘glutathione reaction with an electrophile’ leading to male fertility toxicity. The creation of the AOP provided the mechanistic rationale for the curation of pre-existing structural alerts to relevant key events. Integrating this new knowledge and associated alerts into the DART AOP network will improve its coverage of DART-relevant chemical space. In addition, broadening the coverage of AOPs for a particular regulatory endpoint may facilitate the development of, and confidence in, robust IATAs.

Atlantic herring in International Council for Exploration of the Sea (ICES) Divisions 6.a, 7.b–c comprises at least three populations, distinguished by temporal and spatial differences in spawning, which have until recently been managed as two stocks defined by geographical delineators. Outside of spawning the populations form mixed aggregations, which are the subject of acoustic surveys. The inability to distinguish the populations has prevented the development of separate survey indices and separate stock assessments. A panel of 45 single-nucleotide polymorphisms, derived from whole-genome sequencing, were used to genotype 3480 baseline spawning samples (2014–2021). A temporally stable baseline comprising 2316 herring from populations known to inhabit Division 6.a was used to develop a genetic assignment method, with a self-assignment accuracy greater than 90%. The long-term temporal stability of the assignment model was validated by assigning archive (2003–2004) baseline samples (270 individuals) with a high level of accuracy. Assignment of non-baseline samples (1514 individuals) from Divisions 6.a, 7.b–c indicated previously unrecognized levels of mixing of populations outside of the spawning season. The genetic markers and assignment models presented constitute a ‘toolbox’ that can be used for the assignment of herring caught in mixed survey and commercial catches in Division 6.a into their population of origin with a high level of accuracy.

Mechanically alloyed Fe-based alloys with oxide dispersion strengthening have largely dropped out of the marketplace due to high cost related to problems with complex and unreliable processing. Nevertheless, the desirable properties of oxide dispersion strengthened (ODS) steels have motivated research on alternate processing routes aimed at improving processing simplicity and reliability. Powders produced by gas atomization reaction synthesis (GARS) consist of stable Fe-Y intermetallic phases and a Cr surface oxide layer that acts as a chemical reservoir during solid-state processing and heat treatment to form a high density of nano-scale oxides. This research explores the use of Fe GARS powders, with 15 wt% Cr with micro-alloyed additions of 0.15 wt% Y and 0.10% Ti, in laser powder bed fusion (LPBF) additive manufacturing (AM), and evaluates the effectiveness of oxide dispersoid formation in the liquid melt pool. Additional oxygen was introduced by varying the LPBF chamber atmospheres using Ar, Ar + 1 wt% O, Ar + 5 wt% O, and air. Characterization of LPBF consolidated solids demonstrated the formation of a high density of nano-scale Y-Ti oxides in the build microstructures from the GARS precursor powders.

Among other tests, Barts Health NHS Trust clinical transplantation laboratory conducts two important gene-detection tests: human leucocyte antigen (HLA)-B*27 (‘B27’, associated with the diagnosis of ankylosing spondylitis) and HLA-B*57:01 (‘B57’, associated with prediction of abacavir hypersensitivity disorder). The turnaround time (TaT) from sample receipt to return of results is important to clinicians and their patients but was not monitored. Furthermore, we anticipated an imminent increase in demand from a forthcoming pathology service merger, together with long-term increases with the rise of personalised genetic medicine.In this quality improvement project, we identified current TaT performance and sources of delay. Over three plan-do-study-act (PDSA) cycles, we tested three change ideas, two involving using IT to remove manual administrative steps and alert us to samples needing progressing; both were retained. The other change involved separating out the targeted tests; we judged this not worthwhile with current demand levels, although something to be re-examined when volumes increase. During the project, we reduced mean TaT from 3.8 to 3.3 days and increased the proportion within our 5-day target from 78% to 100%. These have been sustained (at 3.4 days and 97%) for the 3 months following our PDSA cycles and illustrate that reducing variation can be as impactful as reducing the mean.We conducted this project during the COVID-19 disruption, which reduced demand substantially. We took advantage of this to allow staff to spend time on these improvement activities. Another interesting feature of the work is that during the project, we compared changes in performance on our targeted B27/B57 tests with that on another comparable test as a control, to consider the impact of the general increased attention (the Hawthorne effect). We found that performance on this control also increased comparably, but then fell away after our project finished, while it did not for B27/B57.

Abstract Objective Obesity is associated with a high incidence of obstructive sleep apnoea (OSA). Bariatric surgery is postulated to lead to OSA resolution, but there is inconclusive evidence on its efficacy. We used objective measurements to determine the rate of resolution or improvement of OSA in patients who had bariatric procedures in our unit. Results Data was analysed on all patients with OSA who underwent bariatric procedures [laparoscopic Roux-en-Y gastric bypass (LRYGB) and sleeve gastrectomy (LSG)] between June 2012 and September 2016 in our unit. 47 patients (26.7%) were diagnosed with OSA. Mean age was 48.5 years. 63.8% were female. 43 required nocturnal continuous positive airway pressure (CPAP) support. Procedures were LRYGB (n = 26) and LSG (n = 21). Mean excess weight loss was 56.1%. Mean start apnoea-hypopnoea index (AHI) on CPAP was 6.4 events/hr and end AHI was 1.4 events/h. 14 patients (32.6%) had complete OSA resolution and 12 (27.9%) showed improvement in pressure support requirements. We demonstrated that 55.3% of patients had resolution or improvement in OSA following bariatric surgery. However, there was a high rate of non-attendance of follow-up appointments. Future efforts will involve analysis of the reasons for this to ensure more robust monitoring.

Purpose: With the increasing burden of chronic pain and opioid use, provider shortages in Eastern Kentucky and West Virginia have experienced many challenges related to chronic pain management. This study tested a practice facilitator model in both academic and community clinics that selected and implemented best practice processes to better assist patients with chronic pain and increase the use of interdisciplinary health care services. Methods: Using a quasi-experimental design, a practice facilitator was assigned to each state’s clinics and trained clinic teams in quality improvement methods to implement chronic pain tool(s) and workflow processes. Charts for 695 patients with chronic pain using opioids, from 8 randomly selected clinics in eastern Appalachia, were reviewed to assess for changes in clinic processes. Results: Statistically significant improvements were found in 10 out of 16 chronic pain best practice process measures. These included improved workflow implementation (P < 0.001), increased urine drug screen test orders (P = 0.001) and increased utilization of controlled medication agreements (P = 0.004). In total, 7 of 8 clinics significantly improved in at least one, if not all, selected and implemented process measures. Conclusions: Our findings indicate that practice facilitation, standardization of workflows and formation of structured clinical teams can improve processes of care in chronic pain management and facilitate the use of interdisciplinary services. Future studies are needed to assess long-term patient-centered outcomes that may result from improved processes of chronic pain care.

ABSTRACT Objectives The effective exploitation of what are often called big data is increasingly important. They provide the evidence in evidence-based health care and underpin scientific progress in many domains including social/economic policy. Typically, an optimal analysis involves working directly with microdata; i.e. the detailed data relating to each individual in the dataset. But there are many ethico-legal and other governance restrictions on physically sharing microdata. Furthermore, researchers or institutions may have an extensive intellectual property investment in complex microdata and although keen for other researchers to analyse their data they may not wish to give them a physical copy. These restrictions can discourage the use of optimum approaches to analysing pivotal data and slow scientific progress. Data science groups across the world are exploring privacy-protected approaches to analysing microdata without having to physically share the data. Approach A two day international workshop was arranged focussing on privacy protected approaches to data analysis – particularly federated analysis where raw data remain at their original site of collection. The workshop considered the range of approaches that exist, and those that are currently being developed. It explored the strengths, weaknesses, opportunities and challenges associated with these methods and identified situations where specific approaches have a particularly important role. The workshop included a number of practical sessions where potential users could watch demonstrations of the various approaches in action and run analyses themselves. Results The Data Analysis with Privacy Protection for Epidemiological Research (DAPPER) workshop was held 22-23rd August 2016, Bristol. We report back to the broader community on the outcomes of this workshop that focussed on exploring current approaches, tools and technical solutions that facilitate sensitive data to be shared and analysed. Conclusions The workshop has helped map out key opportunities and challenges and assisted potential users, developers and other stakeholders (e.g. funders/journals) to recognise the strengths and weaknesses of different privacy protected analytic approaches. The workshop will encourage further methodological work in this field and better informed application of existing methods.

We investigated the visual scanning strategy employed by a group of individuals with a severe traumatic brain injury (TBI) during a facial affect recognition task. Four males with a severe TBI were matched for age and gender with 4 healthy controls. Eye movements were recorded while pictures of static emotional faces were viewed (i.e., sad, happy, angry, disgusted, anxious, surprised). Groups were compared with respect to accuracy in labelling the emotional facial expression, reaction time, number and duration of fixations to internal (i.e., eyes + nose + mouth), and external (i.e., all remaining) regions of the stimulus. TBI participants demonstrated significantly reduced accuracy and increased latency in facial affect recognition. Further, they demonstrated no significant difference in the number or duration of fixations to internal versus external facial regions. Control participants, however, fixated more frequently and for longer periods of time upon internal facial features. Impaired visual scanning can contribute to inaccurate interpretation of facial expression and this can disrupt interpersonal communication. The scanning strategy demonstrated by our TBI group appears more ‘widespread’ than that employed by their normal counterparts. Further work is required to elucidate the nature of the scanning strategy used and its potential variance in TBI.

Introduction Mental well-being is a global public health priority with increasing mental health conditions having substantial burden on individuals, health systems and society. ‘Stepped care’, where services are provided at an intensity to meet the changing needs of the consumer, is the chosen approach to mental health service delivery in primary healthcare in Australia for its efficiencies and patient outcomes; yet limited evidence exists on how the programme is being rolled out and its impact in practice. This protocol outlines a data linkage project to characterise and quantify healthcare service utilisation and impacts among a cohort of consumers of a national mental health stepped care programme in one region of Australia.Methods and analysis Data linkage will be used to establish a retrospective cohort of consumers of mental health stepped care services between 1 July 2020 and 31 December 2021 in one primary healthcare region in Australia (n=approx. 12 710). These data will be linked with records from other healthcare service data sets (eg, hospitalisations, emergency department presentations, community-based state government-delivered mental healthcare, hospital costs). Four areas for analysis will include: (1) characterising the nature of mental health stepped care service use; (2) describing the cohort’s sociodemographic and health characteristics; (3) quantifying broader service utilisation and associated economic costs; and (4) assessing the impact of mental health stepped care service utilisation on health and service outcomes.Ethics and dissemination Approval from the Darling Downs Health Human Research Ethics Committee (HREA/2020/QTDD/65518) has been granted. All data will be non-identifiable, and research findings will be disseminated through peer-reviewed journals, conference presentations and industry meetings.

Abstract Providing electronic health data to medical practitioners to reflect on their performance can lead to improved clinical performance and quality of care. Understanding the sensemaking process that is enacted when practitioners are presented with such data is vital to ensure an improvement in performance. Thus, the primary objective of this research was to explore physician and surgeon sensemaking when presented with electronic health data associated with their clinical performance. A systematic literature review was conducted to analyse qualitative research that explored physicians and surgeons experiences with electronic health data associated with their clinical performance published between January 2010 and March 2022. Included articles were assessed for quality, thematically synthesised, and discussed from the perspective of sensemaking. The initial search strategy for this review returned 8,829 articles that were screened at title and abstract level. Subsequent screening found 11 articles that met the eligibility criteria and were retained for analyses. Two articles met all of the standards within the chosen quality assessment (Standards for Reporting Qualitative Research, SRQR). Thematic synthesis generated five overarching themes: data communication, performance reflection, infrastructure, data quality, and risks. The confidence of such findings is reported using CERQual (Confidence in the Evidence from Reviews of Qualitative research). The way the data is communicated can impact sensemaking which has implications on what is learned and has impact on future performance. Many factors including data accuracy, validity, infrastructure, culture can also impact sensemaking and have ramifications on future practice. Providing data in order to support performance reflection is not without risks, both behavioural and affective. The latter of which can impact the practitioner’s ability to effectively make sense of the data. An important consideration when data is presented with the intent to improve performance. Registration This systematic review was registered with Prospero, registration number: CRD42020197392.

BackgroundHospitals routinely collect large amounts of administrative data such as length of stay, 28-day readmissions, and hospital-acquired complications; yet, these data are underused for continuing professional development (CPD). First, these clinical indicators are rarely reviewed outside of existing quality and safety reporting. Second, many medical specialists view their CPD requirements as time-consuming, having minimal impact on practice change and improving patient outcomes. There is an opportunity to build new user interfaces based on these data, designed to support individual and group reflection. Data-informed reflective practice has the potential to generate new insights about performance, bridging the gap between CPD and clinical practice. ObjectiveThis study aims to understand why routinely collected administrative data have not yet become widely used to support reflective practice and lifelong learning. MethodsWe conducted semistructured interviews (N=19) with thought leaders from a range of backgrounds, including clinicians, surgeons, chief medical officers, information and communications technology professionals, informaticians, researchers, and leaders from related industries. Interviews were thematically analyzed by 2 independent coders. ResultsRespondents identified visibility of outcomes, peer comparison, group reflective discussions, and practice change as potential benefits. The key barriers included legacy technology, distrust with data quality, privacy, data misinterpretation, and team culture. Respondents suggested recruiting local champions for co-design, presenting data for understanding rather than information, coaching by specialty group leaders, and timely reflection linked to CPD as enablers to successful implementation. ConclusionsOverall, there was consensus among thought leaders, bringing together insights from diverse backgrounds and medical jurisdictions. We found that clinicians are interested in repurposing administrative data for professional development despite concerns with underlying data quality, privacy, legacy technology, and visual presentation. They prefer group reflection led by supportive specialty group leaders, rather than individual reflection. Our findings provide novel insights into the specific benefits, barriers, and benefits of potential reflective practice interfaces based on these data sets. They can inform the design of new models of in-hospital reflection linked to the annual CPD planning-recording-reflection cycle.

'CLIMATE CHANGE is the most severe problem we are facing today--more serious even than the threat of terrorism,' according to the government's chief scientist, David King. This might sound alarmist, [...]

This paper assesses the effectiveness of essay tutorials offered to first-year economics students at the UCT (University of Cape Town) in their first language (L1). All students in the study are first-language speakers of an African language. Firstly, using propensity score matching, we econometrically assess the impact of these tutorials on students’ essay marks. Although our sample size is small [n=220], our findings provide preliminary evidence of a positive impact of the intervention on a student’s final essay mark. The results show that the average gain for students who attended an essay tutorial in their L1 was 4.85%, with this result being statistically significant at the 10% significance level. Secondly, students’ perceptions of the tutorials’ effectiveness, as documented by online evaluations and focus groups, are examined. These findings suggest that allowing for unmediated L1 use in tertiary education classrooms can foster inclusivity and promote participation in otherwise largely monolingual spaces.

# Background Diabetes mellitus (DM) is a global public health concern, with significant increase in the transitioning low- and middle-income countries (LMICs), such as India, with limited resources and poor health care systems. Recognising the need for innovative, feasible, targeted interventions for diabetes care, a non-randomized, controlled, phase II clinical trial was conducted in a rural revenue block in Tamil Nadu, India. This pilot trial tested the non-inferiority of the trained community health workers (CHWs) in delivering education, screening for diabetes complications using a protocol-based referral for physician consultation for management, assisted by an android application (App) for patient tracking. Understanding the perceptions of key stakeholders including patients and health workers is crucial for upscaling and sustaining these novel interventions for diabetes care. This study explored the perspectives and views of patients with DM and diabetes CHWs who participated in the pilot trial. # Methods Using qualitative research methodology, individual in-depth interviews were conducted in the local vernacular (Tamil) language among the two diabetes CHWs and purposively chosen 24 patients until the data saturation was reached. Participant information sheets, consent forms and a detailed topic guide were developed. Interviews were transcribed verbatim, translated into English and thematic analysis was conducted. # Results Five main themes emerged during interviews with diabetes CHWs: deeper understanding of the disease; belonging and strengthening; ease of use of digital technology and a timesaver for patient tracking; cultural sensitivity; stigma and change in attitudes over time. Interviews with patients highlighted four major themes: convenience; cost reduction; improved knowledge and perceptions on health. Benefits were perceived by both patients and diabetes CHWs, in the domains of convenience, access, cost and knowledge. Perceptions of changes in health were mixed with room for improvement in the digital App. # Conclusions Training CHWs in resource limited settings in supporting community diabetes care, assisted by digital technology for tracking is acceptable to the patients and will help shift the care pathway for Diabetes closer to the patients in a rural setting. This qualitative study identified a number of areas for improvement and potential barriers that will need to be addressed prior to scaling up in similar resource constrained settings.

Summary: Reproducibility in molecular and cellular studies is fundamental to scientific discovery. To establish the reproducibility of a well-defined long-term neuronal differentiation protocol, we repeated the cellular and molecular comparison of the same two iPSC lines across five distinct laboratories. Despite uncovering acceptable variability within individual laboratories, we detect poor cross-site reproducibility of the differential gene expression signature between these two lines. Factor analysis identifies the laboratory as the largest source of variation along with several variation-inflating confounders such as passaging effects and progenitor storage. Single-cell transcriptomics shows substantial cellular heterogeneity underlying inter-laboratory variability and being responsible for biases in differential gene expression inference. Factor analysis-based normalization of the combined dataset can remove the nuisance technical effects, enabling the execution of robust hypothesis-generating studies. Our study shows that multi-center collaborations can expose systematic biases and identify critical factors to be standardized when publishing novel protocols, contributing to increased cross-site reproducibility. : In this article, Lakics and colleagues show that, while individual laboratories are able to identify consistent molecular and seemingly statistically robust differences between iPSC neuronal models, cross-site reproducibility is poor. Their findings support multi-center collaborations to expose systematic biases and identify critical factors to be standardized to improve reproducibility in iPSC-based molecular experiments. Keywords: induced pluripotent stem cell, reproducibility, cross-site experimental variation, cortical neurons, gene expression profile, proteomic profiles, single-cell sequencing, molecular profiling, stembancc, public-private partnership

Knowledge Translation (KT) is increasingly a requirement for scholars and non-academics working in applied settings. However, few programs provide explicit training in KT. In this article we systematically explore our experiences as a multi-disciplinary group of course facilitators and students in a newly redeveloped graduate course in Evidence Based Practice and Knowledge Translation. The course was designed to emphasize handson learning, collaboration and community engagement. We reflect on the challenges we faced and the skills, knowledge and opportunities that students gained as they developed and implemented community-based KT strategies relating to refugee resettlement, young carers, and consumer attitudes, behaviour and values around food purchasing decisions. We conclude by providing recommendations for instructors and institutions for implementing learning experiences in KT that are designed for real-world impact. L’application des connaissances (AC) est devenue une exigence de plus en plus fréquente pour les chercheurs et les personnes qui travaillent dans les milieux non universitaires. Toutefois, peu de programmes offrent une formation explicite en AC. Dans cet article, nous explorons systématiquement nos expériences en tant que groupe pluridisciplinaire formé de responsables de cours et d’étudiants dans un cours de cycle supérieur nouvellement remanié portant sur la pratique fondée sur les données probantes et l’application des connaissances. Le cours a été conçu pour mettre en valeur l’apprentissage pratique, la collaboration et l’engagement communautaire. Nous réfléchissons aux défis auxquels nous avons été confrontés ainsi qu’aux compétences, aux connaissances et aux opportunités que les étudiants ont acquis en développant et mettant en pratique des stratégies d’AC en milieu communautaire sur les thèmes de la réinstallation des réfugiés, des jeunes aidants et des attitudes, comportements et valeurs des consommateurs en matière d’achat de produits alimentaires. En conclusion, nous présentons des recommandations à l’intention des enseignants et des établissements pour la mise en pratique d’expériences en AC qui soient conçues pour avoir un effet dans le monde réel.

We previously identified Wiz in a mouse screen for epigenetic modifiers. Due to its known association with G9a/GLP, Wiz is generally considered a transcriptional repressor. Here, we provide evidence that it may also function as a transcriptional activator. Wiz levels are high in the brain, but its function and direct targets are unknown. ChIP-seq was performed in adult cerebellum and Wiz peaks were found at promoters and transcription factor CTCF binding sites. RNA-seq in Wiz mutant mice identified genes differentially regulated in adult cerebellum and embryonic brain. In embryonic brain most decreased in expression and included clustered protocadherin genes. These also decreased in adult cerebellum and showed strong Wiz ChIP-seq enrichment. Because a precise pattern of protocadherin gene expression is required for neuronal development, behavioural tests were carried out on mutant mice, revealing an anxiety-like phenotype. This is the first evidence of a role for Wiz in neural function.

Emma Whitehead is an alumna of the Center for Partnership Studies Young Women's Leadership Program. In this article she reflects on her experience as a delegate to the 2016 United States Naval Academy Foreign Affairs Conference. The conference promoted the status of women and girls globally and Whitehead participated in a round table session on gender equity, sharing insights from the Center for Partnership Studies.

Transposable elements (TEs) have been active in the mammalian genome for millions of years and the silencing of these elements in the germline is important for the survival of the host. Mice carrying reporter transgenes can be used to model transcriptional silencing. A mutagenesis screen for modifiers of epigenetic gene silencing produced a line with a mutation in Trim33; the mutants displayed increased expression of the reporter transgene. ChIP-seq of Trim33 in testis revealed 9,109 peaks, mostly at promoters. This is the first report of ChIP-seq for Trim33 in any tissue. Comparison with ENCODE datasets showed that regions of high read density for Trim33 had high read density for histone marks associated with transcriptional activity and mapping to TE consensus sequences revealed Trim33 enrichment at RLTR10B, the LTR of one of the youngest retrotransposons in the mouse genome, MMERVK10C. We identified consensus sequences from the 266 regions at which Trim33 ChIP-seq peaks overlapped RLTR10B elements and found a match to the A-Myb DNA-binding site. We found that TRIM33 has E3 ubiquitin ligase activity for A-MYB and regulates its abundance. RNA-seq revealed that mice haploinsufficient for Trim33 had altered expression of a small group of genes in the testis and the gene with the most significant increase was found to be transcribed from an upstream RLTR10B. These studies provide the first evidence that A-Myb has a role in the actions of Trim33 and suggest a role for both A-Myb and Trim33 in the arms race between the transposon and the host. This the first report of any factor specifically regulating RLTR10B and adds to the current literature on the silencing of MMERVK10C retrotransposons. This is also the first report that A-Myb has a role in the transcription of any retrotransposon.

Previous studies on the epigenetic regulator DNA methyltransferase 3-Like (DNMT3L), have demonstrated it is an essential regulator of paternal imprinting and early male meiosis. Dnmt3L is also a paternal effect gene, i.e., wild type offspring of heterozygous mutant sires display abnormal phenotypes suggesting the inheritance of aberrant epigenetic marks on the paternal chromosomes. In order to reveal the mechanisms underlying these paternal effects, we have assessed X chromosome meiotic compaction, XY chromosome aneuploidy rates and global transcription in meiotic and haploid germ cells from male mice heterozygous for Dnmt3L. XY bodies from Dnmt3L heterozygous males were significantly longer than those from wild types, and were associated with a three-fold increase in XY bearing sperm. Loss of a Dnmt3L allele resulted in deregulated expression of a large number of both X-linked and autosomal genes within meiotic cells, but more prominently in haploid germ cells. Data demonstrate that similar to embryonic stem cells, DNMT3L is involved in an auto-regulatory loop in germ cells wherein the loss of a Dnmt3L allele resulted in increased transcription from the remaining wild type allele. In contrast, however, within round spermatids, this auto-regulatory loop incorporated the alternative non-coding alternative transcripts. Consistent with the mRNA data, we have localized DNMT3L within spermatids and sperm and shown that the loss of a Dnmt3L allele results in a decreased DNMT3L content within sperm. These data demonstrate previously unrecognised roles for DNMT3L in late meiosis and in the transcriptional regulation of meiotic and post-meiotic germ cells. These data provide a potential mechanism for some cases of human Klinefelter's and Turner's syndromes.

Recent studies have shown that exposure to some nutritional supplements and chemicals in utero can affect the epigenome of the developing mouse embryo, resulting in adult disease. Our hypothesis is that epigenetics is also involved in the gestational programming of adult phenotype by alcohol. We have developed a model of gestational ethanol exposure in the mouse based on maternal ad libitum ingestion of 10% (v/v) ethanol between gestational days 0.5-8.5 and observed changes in the expression of an epigenetically-sensitive allele, Agouti viable yellow (A(vy)), in the offspring. We found that exposure to ethanol increases the probability of transcriptional silencing at this locus, resulting in more mice with an agouti-colored coat. As expected, transcriptional silencing correlated with hypermethylation at A(vy). This demonstrates, for the first time, that ethanol can affect adult phenotype by altering the epigenotype of the early embryo. Interestingly, we also detected postnatal growth restriction and craniofacial dysmorphology reminiscent of fetal alcohol syndrome, in congenic a/a siblings of the A(vy) mice. These findings suggest that moderate ethanol exposure in utero is capable of inducing changes in the expression of genes other than A(vy), a conclusion supported by our genome-wide analysis of gene expression in these mice. In addition, offspring of female mice given free access to 10% (v/v) ethanol for four days per week for ten weeks prior to conception also showed increased transcriptional silencing of the A(vy) allele. Our work raises the possibility of a role for epigenetics in the etiology of fetal alcohol spectrum disorders, and it provides a mouse model that will be a useful resource in the continued efforts to understand the consequences of gestational alcohol exposure at the molecular level.

There is increasing evidence in both plants and animals that epigenetic marks are not always cleared between generations. Incomplete erasure at genes associated with a measurable phenotype results in unusual patterns of inheritance from one generation to the next, termed transgenerational epigenetic inheritance. The Agouti viable yellow (A(vy)) allele is the best-studied example of this phenomenon in mice. The A(vy) allele is the result of a retrotransposon insertion upstream of the Agouti gene. Expression at this locus is controlled by the long terminal repeat (LTR) of the retrotransposon, and expression results in a yellow coat and correlates with hypomethylation of the LTR. Isogenic mice display variable expressivity, resulting in mice with a range of coat colours, from yellow through to agouti. Agouti mice have a methylated LTR. The locus displays epigenetic inheritance following maternal but not paternal transmission; yellow mothers produce more yellow offspring than agouti mothers. We have analysed the DNA methylation in mature gametes, zygotes, and blastocysts and found that the paternally and maternally inherited alleles are treated differently. The paternally inherited allele is demethylated rapidly, and the maternal allele is demethylated more slowly, in a manner similar to that of nonimprinted single-copy genes. Interestingly, following maternal transmission of the allele, there is no DNA methylation in the blastocyst, suggesting that DNA methylation is not the inherited mark. We have independent support for this conclusion from studies that do not involve direct analysis of DNA methylation. Haplo-insufficiency for Mel18, a polycomb group protein, introduces epigenetic inheritance at a paternally derived A(vy) allele, and the pedigrees reveal that this occurs after zygotic genome activation and, therefore, despite the rapid demethylation of the locus.