High-throughput methylome analysis reveals differential methylation for early and late onset preeclampsia for mothers and their children.

Preeclampsia is a hypertensive disorder of pregnancy that affects ∼2%-5% of all pregnancies, contributes to 4 of the top 10 causes of pregnancy-related deaths, and remains a long-term risk factor for cardiometabolic diseases. Yet, little is still known about the molecular mechanisms that lead to this disease. There is evidence that some cases have a genetic cause. However, it is well appreciated that harmful factors in the environment, such as poor nutrition, stress, and toxins, may lead to epigenetics changes that can contribute to this disease. DNA methylation is one of the epigenetic modifications known to be fairly stable and impact gene expression. Using DNA from buccal swabs, we analyzed global DNA methylation among three groups of individuals: mothers who experienced 1) early-stage preeclampsia (<32 wk), 2) late-stage preeclampsia (>37 wk), or 3) no complications during their pregnancies, as well as the children from these three groups. We found significant differentially methylated regions (DMRs) between mothers who experienced preeclampsia compared with those with no complications adjacent or within genes that are important for placentation, embryonic development, cell adhesion, and inflammation (e.g., the cadherin pathway). A significant portion of DMR genes showed expression in tissues relevant to preeclampsia (i.e., the brain, heart, kidney, uterus, ovaries, and placenta). As this study was performed on DNA extracted from cheek swabs, this opens the way to future studies in different tissues, aimed at identifying possible biomarkers of risk and early detection, developing targeted interventions, and reducing the progression of this life-threatening disease.NEW & NOTEWORTHY Preeclampsia is a life-threatening hypertensive disorder, affecting 2%-5% of pregnancies, that remains poorly understood. This study analyzed DNA methylation from buccal swabs from mothers who experienced early and late-stage preeclampsia and those with uncomplicated pregnancies, along with their children. Differentially methylated regions were found near and within genes crucial for placental function, embryonic development, and inflammation. Many of these genes are expressed in preeclampsia-related tissues, offering hope for future biomarker development for this condition.

Human-animal interactions and machine-animal interactions in animals under human care: A summary of stakeholder and researcher perceptions and future directions.

Animals under human care are exposed to a potentially large range of both familiar and unfamiliar humans. Human-animal interactions vary across settings, and individuals, with the nature of the interaction being affected by a suite of different intrinsic and extrinsic factors. These interactions can be described as positive, negative or neutral. Across some industries, there has been a move towards the development of technologies to support or replace human interactions with animals. Whilst this has many benefits, there can also be challenges associated with increased technology use. A day-long Animal Welfare Research Network workshop was hosted at Harper Adams University, UK, with the aim of bringing together stakeholders and researchers (n = 38) from the companion, farm and zoo animal fields, to discuss benefits, challenges and limitations of human-animal interactions and machine-animal interactions for animals under human care and create a list of future research priorities. The workshop consisted of four talks from experts within these areas, followed by break-out room discussions. This work is the outcome of that workshop. The key recommendations are that approaches to advancing the scientific discipline of machine-animal interactions in animals under human care should focus on: (1) interdisciplinary collaboration; (2) development of validated methods; (3) incorporation of an animal-centred perspective; (4) a focus on promotion of positive animal welfare states (not just avoidance of negative states); and (5) an exploration of ways that machines can support a reduction in the exposure of animals to negative human-animal interactions to reduce negative, and increase positive, experiences for animals.

Qualitative Behavioural Assessment of bonobo emotional expressivity across observer groups and zoo housing environments.

Human evaluation of animal emotional expressivity can inform animal welfare. Qualitative Behavioural Assessment (QBA) has been applied to domesticated and some non-domesticated animals, but its use in primates is limited despite their emotional expressivity. We aimed to develop and apply a QBA for bonobos (Pan paniscus) through two consecutive studies. We applied Free Choice Profiling (FCP) and the Fixed List methodology, respectively, in Study 1 and 2, and invited students and bonobo experts to rate video clips of zoo-living bonobos of different sexes and age classes, and before and after moving to a new enclosure. In Study 1, students described dimension 1 as ranging from 'quiet/calm' to 'angry/active' and dimension 2 from 'sad/anxious' to 'happy/loving'. Experts described dimension 1 ranging from 'quiet/relaxed' to 'nervous/alert' and dimension 2 from 'nervous/bored' to 'playful/happy'. Using a fixed list of descriptors, informed by findings from Study 1, students in Study 2 described dimension 1 as ranging from 'quiet/calm' to 'agitated/frustrated', and dimension 2 from 'sad/stressed' to 'happy/positively engaged'. Experts described dimension 1 as ranging from 'quiet/calm' to 'active/excited', and dimension 2 from 'sad/bored' to 'happy/positively engaged'. Students scored adults as more 'calm/quiet' and experts scored subadults as more 'happy/positively engaged'. Additionally, experts in Study 2 rated bonobos as more 'active/excited' in their new enclosure. Reliability was moderate to good for the dimensions. Additionally, animal-directed empathy of observers influenced QBA scores. This is the first time, FCP has been successfully used as a method to study primate emotional expressivity. Our findings show the promise of employing QBA in primate studies and in industry, with validation of additional metrics to enable its use for welfare-monitoring purposes.

Molecular identification of hymenopteran parasitoids and their endosymbionts from agromyzids.

Three polyphagous pest Liriomyza spp. (Diptera: Agromyzidae) have recently invaded Australia and are damaging horticultural crops. Parasitic wasps are recognized as effective natural enemies of leafmining species globally and are expected to become important biocontrol agents in Australia. However, the hymenopteran parasitoid complex of agromyzids in Australia is poorly known and its use hindered due to taxonomic challenges when based on morphological characters. Here, we identified 14 parasitoid species of leafminers based on molecular and morphological data. We linked DNA barcodes (5' end cytochrome c oxidase subunit I (COI) sequences) to five adventive eulophid wasp species (Chrysocharis pubicornis (Zetterstedt), Diglyphus isaea (Walker), Hemiptarsenus varicornis (Girault), Neochrysocharis formosa (Westwood), and Neochrysocharis okazakii Kamijo) and two braconid species (Dacnusa areolaris (Nees) and Opius cinerariae Fischer). We also provide the first DNA barcodes (5' end COI sequences) with linked morphological characters for seven wasp species, with three identified to species level (Closterocerus mirabilis Edwards & La Salle, Trigonogastrella parasitica (Girault), and Zagrammosoma latilineatum Ubaidillah) and four identified to genus (Aprostocetus sp., Asecodes sp., Opius sp. 1, and Opius sp. 2). Phylogenetic analyses suggest C. pubicornis, D. isaea, H. varicornis, and O. cinerariae are likely cryptic species complexes. Neochrysocharis formosa and Aprostocetus sp. specimens were infected with Rickettsia. Five other species (Cl. mirabilis, D. isaea, H. varicornis, Opius sp. 1, and Opius sp. 2) were infected with Wolbachia, while two endosymbionts (Rickettsia and Wolbachia) co-infected N. okazakii. These findings provide background information about the parasitoid fauna expected to help control the leafminers.

Co-option of the lineage-specific LAVA retrotransposon in the gibbon genome.

Co-option of transposable elements (TEs) to become part of existing or new enhancers is an important mechanism for evolution of gene regulation. However, contributions of lineage-specific TE insertions to recent regulatory adaptations remain poorly understood. Gibbons present a suitable model to study these contributions as they have evolved a lineage-specific TE called LAVA (LINE-AluSz-VNTR-AluLIKE), which is still active in the gibbon genome. The LAVA retrotransposon is thought to have played a role in the emergence of the highly rearranged structure of the gibbon genome by disrupting transcription of cell cycle genes. In this study, we investigated whether LAVA may have also contributed to the evolution of gene regulation by adopting enhancer function. We characterized fixed and polymorphic LAVA insertions across multiple gibbons and found 96 LAVA elements overlapping enhancer chromatin states. Moreover, LAVA was enriched in multiple transcription factor binding motifs, was bound by an important transcription factor (PU.1), and was associated with higher levels of gene expression in cis We found gibbon-specific signatures of purifying/positive selection at 27 LAVA insertions. Two of these insertions were fixed in the gibbon lineage and overlapped with enhancer chromatin states, representing putative co-opted LAVA enhancers. These putative enhancers were located within genes encoding SETD2 and RAD9A, two proteins that facilitate accurate repair of DNA double-strand breaks and prevent chromosomal rearrangement mutations. Co-option of LAVA in these genes may have influenced regulation of processes that preserve genome integrity. Our findings highlight the importance of considering lineage-specific TEs in studying evolution of gene regulatory elements.

Furred and feathered friends: How attached are zookeepers to the animals in their care?

Keeper-animal relationships (KARs) appear to be important in zoos, since they can enhance the well-being of both the animals and the keepers, can make animal husbandry easier, but conversely might risk inappropriate habituation of animals and possible risks to the safety of keepers. It is, therefore, important to know more about the variables involved in relationship formation. Here we use a modified version of the Lexington Attachment to Pets Scale (LAPS) to measure the strength of KARs between keepers and animals in their care, both in the zoo and in the home. LAPS questionnaires were completed by 187 keepers in 19 different collections across three countries. LAPS scores for attachment to zoo animals (ZA) were significantly lower than for pet animals (PA). There was no significant difference in ZA scores between different taxa, but there were significant taxon differences between PA scores. There were significant differences in both ZA and PA scores between different collections. Female respondents scored more highly than males for both ZA and PA. Multiple regression revealed that location, gender, and time spent with animals were significant predictors for ZA, while only gender and taxon were significant predictors for PA. It was concluded that PA scores were comparable with those for the general public, and reflected strong attachment of keepers to their pets, while ZA scores, although also reflecting attachment, were influenced by differences in institutional culture.

Understanding impacts of zoo visitors: Quantifying behavioural changes of two popular zoo species during COVID-19 closures.

Visitors are normally a prominent and constant feature in a zoo animals' environment with more than 700 million people visiting zoos and aquariums worldwide, annually. Animal-visitor interactions can be enriching and stimulating and are now considered within the Five Domains of animal welfare assessment. Zoo closures as a result of COVID-19 provided a unique opportunity to monitor the impact of abrupt and prolonged removal of visitors on two popular zoo species. Data were collected at four facilities (n = 3 slender-tailed meerkats, n = 1 African penguin) during COVID-19 zoo closures and up to one month following reopening to the public. Meerkats showed increased positive social interactions, increased alert behaviours, and reduced environmental interactions in the first month post-opening, as compared to closure periods. They also used more of their enclosures during periods of closure and spent longer than would be expected in zones furthest from visitor viewing areas when facilities reopened. African penguins showed no behavioural change between open and closure periods. Enclosure usage during both observation periods was relatively even and no differences were observed in enclosure use between open and closure periods. These results will enable an advanced understanding of the impact that people have on the behaviour of zoo animals, which has ramifications for animals used in close encounters and other 'visitor experiences' in the future. Understanding relationships between animals and people is applicable in all managed animal settings. The results from this study are of practical use in managing visitor access to animals moving forwards, including enclosure location and design, to ensure a positive visitor experience that does not negatively impact animal behaviour.

Visitor attachment to dolphins during an interaction programme, are there implications to dolphin behavior?

Millions of people visit zoos and aquariums globally each year, with a smaller number choosing to participate in animal interaction programmes which allows visitors closer contact with individual animals. These are reportedly having mixed effects in increasing conservation-related behaviors. Human-animal interactions (HAIs) during these programmes are generally positive experiences for the human participants, however are there behavioral implications for the animals involved? The Bottlenose Dolphin (Tursiops truncatus) is the most widely used cetacean for dolphin interactions, known as "swim with dolphin" (SWD) programmes. This study investigated visitor attachment to the dolphins they interacted with, whilst assessing behavioral implications of the dolphins. A total of 41 visitors to a Spanish dolphinarium, who participated in a SWD were surveyed using a modified version of the Lexington Attachment to Pets Scale. Alongside this, 15-min continuous focal samples monitored three female dolphins (D1, D2, and D3) aged 22-40, split into pre (n = 96), during (n = 96) and post (n = 96) SWD. 80% of visitors reported a sense of attachment to the dolphin they interacted with. An exploratory factor analysis extracted three factors from the survey, these were "relationships," "emotional attachment," and "non-attachment." A Friedmans two-way analysis of variance produced significant results for some behavior categories for each individual, including locomotory (D1: F2 = 9.556, p < .01), rest (D2: F2 = 14, p < .01, D3: F2 = 10.889, p < .01) and individual play (D1: F2 = 11.677, p < .01 D2: F2 = 6.353, p < .05) however, pairwise comparison showed no differences pre-post SWD. In this context it can be implied that participating in the SWD was neither enriching nor aversive for the individual animals, although due to the small sample size further research is required. As visitors reported a sense of attachment post HAI, this can have applications in improving conservation education during SWD. This study has provided scope for further research into methods that facilities can use to utilize the emotional attachment developed to individual animals to facilitate learning about conservation issues.

Zoo-housed mammals do not avoid giving birth on weekends.

There is evidence that zoo visitor presence can influence the behaviour and, in some cases, adrenal response of zoo animals, and can sometimes compromise animal welfare. In some laboratory studies, significantly more primate births have been reported on weekends, when fewer people are working there, compared with weekdays when staffing levels are at their highest. Here, we investigate whether there is evidence of a "weekend effect" on births in zoo animals as a result of visitor numbers. Unlike laboratories, zoos are typically busier with visitors on weekends than on weekdays, although staffing levels remain fairly consistent across days of the week. If zoo animal parturition is sensitive to human presence, then fewer births would be expected on weekends compared with weekdays. We tested this using birth data and visitor numbers on the entrance gate from zoo records across 16 species representing artiodactyls, perissodactyls, carnivores and primates at four British zoos, to see whether there is an association between mean daily birth rates and average visitor numbers. We predict that, if there is a visitor effect, daily births should be lower on weekends than weekdays and should correlate with mean daily visitor numbers. Results showed that births for all 16 species were randomly distributed through the week, and there was no significant decline in births on weekends. We conclude that the "weekend effect", if such a thing exists, does not appear to be a feature of zoo births, suggesting that elevated weekend visitor numbers are not sufficiently stressful to trigger delayed parturition.

Is wounding aggression in zoo-housed chimpanzees and ring-tailed lemurs related to zoo visitor numbers?

Chimpanzees in laboratory colonies experience more wounds on weekdays than on weekends, which has been attributed to the increased number of people present during the week; thus, the presence of more people was interpreted as stressful. If this were also true for primates in zoos, where high human presence is a regular feature, this would clearly be of concern. Here we examine wounding rates in two primate species (chimpanzees Pan troglodytes and ring-tailed lemurs Lemur catta) at three different zoos, to determine whether they correlate with mean number of visitors to the zoo. Wounding data were obtained from a zoo electronic record keeping system (ZIMS™). The pattern of wounds did not correlate with mean gate numbers for those days for either species in any group. We conclude that there is no evidence that high visitor numbers result in increased woundings in these two species when housed in zoos. Zoo Biol. 35:205-209, 2016. © 2016 Wiley Periodicals, Inc.

The evolving story of sulfoxaflor resistance in the green peach aphid, Myzus persicae (Sulzer).

BACKGROUND: The green peach aphid, Myzus persicae (Sulzer), is one of the most economically important crop pests worldwide. Insecticide resistance in this pest was first detected over 60 years ago, with resistance in M. persicae now spanning over 80 active ingredients. Sulfoxaflor is a relatively new insecticide that is primarily used to control sap-feeding insects. In 2018 resistance to sulfoxaflor was discovered in field populations of M. persicae in Australia. This study aimed to determine the current distribution and phenotypic levels of sulfoxaflor resistance in Australian clones of M. persicae and to investigate how these patterns relate to clonal type. RESULTS: For the first time, we show there is low-level resistance (8-26-fold) distributed across Australia, with resistance being detected in aphids collected from approximately 20% of all M. persicae collected and screened. Furthermore, this study shows sulfoxaflor resistance is found in two M. persicae haplotypes, providing evidence that there have been multiple independent evolutionary events which have given rise to sulfoxaflor resistance in this species. CONCLUSION: These findings have important implications for the chemical control of M. persicae in Australia, especially when considering the broader genetic background of these aphids which are known to harbour a number of other insecticide resistance mechanisms. We recommend continuous monitoring of sulfoxaflor resistance in field populations of M. persicae (in Australia and elsewhere) and further research into the underlying genetic mechanisms conferring resistance to sulfoxaflor in both clonal haplotypes. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The effects of insecticide seed treatments on the parasitism and predation of Myzus persicae (Homoptera: Aphididae) in canola.

The green peach aphid, Myzus persicae (Sulzer) (Homoptera: Aphididae), is a major pest of brassica plants, with the ability to transmit > 100 viruses. Although the adoption of Integrated Pest Management is increasing, chemical treatment remains the predominant method used to control M. persicae globally. Insecticide seed treatments, typically with neonicotinoid active ingredients, have become commonplace in canola crops, and are viewed as a "softer" alternative to foliar sprays but may nevertheless impact natural enemies of M. persicae. In this study, the effects of canola seed treatments, containing imidacloprid, thiamethoxam, and a mixture of thiamethoxam + lambda-cyhalothrin, were investigated on the parasitoid wasp, Aphidius colemani Viereck (Hymenoptera: Braconidae) and the green lacewing, Mallada signatus (Schneider) (Neuroptera: Chrysopidae), both important natural enemies of M. persicae. Laboratory trials were undertaken using whole plants, with lethal and sublethal effects assessed by measuring several traits. Compared with untreated plants, more aphid mummies were produced and more A. colemani were reared on plants treated with thiamethoxam + lambda-cyhalothrin and more aphid mummies were produced on imidacloprid plants. Imidacloprid reduced the time A. colemani spent searching for M. persicae and thiamethoxam reduced its cleaning time. However, after A. colemani were removed from treated plants, there were no such effects observed, suggesting these impacts were relatively short-lived. We found no significant effects of seed treatments on M. signatus. These results point to the complexity of ecotoxicology studies involving multiple trophic levels and indicate that seed treatments may have variable impacts on key fitness traits of natural enemies.

Current practice of placental cord insertion documentation in Australia - A sonographer survey.

Introduction: During pregnancy, the umbilical cord attaches to the placenta in a central, eccentric, marginal or velamentous location. Maternal and fetal complications are associated with marginal and velamentous cord insertions, the most clinically significant being perinatal mortality due to undiagnosed vasa praevia. Current literature describes a wide variation regarding regulation of placental cord insertion (PCI) documentation during antenatal ultrasound examinations. This prospective cross-sectional study aimed to assess the current practice of antenatal PCI documentation in Australia. Methods: Members of the Australian Sonographer Accreditation Registry were invited to participate in an online survey which was distributed between February and March 2022. Results: Four hundred ninety sonographers met the inclusion criteria for the study of which 330 (67.3%) have more than 10 years' experience as a sonographer and 375 (76.5%) are employed primarily in a public or private setting offering general ultrasound. Most respondents (89.6%) indicated documentation of the PCI site is departmental protocol at the second trimester anatomy scan (17-22 weeks gestation), but PCI documentation is protocol in less than 50% of other obstetric ultrasound examinations listed in the survey. The PCI site is included in the formal ultrasound report at a rate significantly less than inclusion in the departmental protocol and the sonographer's worksheet. Conclusions: Considering the potential maternal and fetal complications associated with abnormal PCI and the ease at which the PCI site is identified in the first and second trimesters, we believe that standard inclusion of the PCI site in departmental protocol and in the formal ultrasound report from 11 weeks gestation, regardless of whether it is normal or abnormal, would prove invaluable.

The Impact of Visitors on Non-Primate Species in Zoos: A Quantitative Review.

Visitors are a prominent feature in the lives of zoo animals, and their presence can cause a range of impacts on zoo animals (typically classed as positive, negative or neutral impacts), commonly referred to as the 'visitor effect'. This paper quantitatively collates the literature on the visitor effect in non-primate species, investigates the types of measures used to assess impacts of visitors on animals and considers whether impacts vary across non-primate species in zoos. In total, there were 105 papers which had investigated the impact of zoo visitors on 252 non-primate species/species groups. There has been a steady increase in visitor effect research in zoos since 2012 and this body of work incorporates species from avian (28% study species), reptilian (9%), amphibian (2%), fish (4%) and invertebrate taxa (1%). However, there is still a bias towards mammalian species (56%). The response to visitors varied across taxa. Amphibians responded negatively to visitors more frequently than would be expected by chance (p < 0.05), birds responded neutrally more frequently than would be expected by chance (p < 0.05) and fish responded neutrally and 'unknown' more frequently than would be expected by chance (p < 0.05). This review highlighted a number of animal-based metrics which have been used to assess the impacts of visitors on animals, with measures used varying across taxa. Moving forwards, it is recommended that moving forwards researchers incorporate a suite of measures, incorporating those which are meaningful in terms of being representative of individual animal experiences and animal welfare, collected in a manner which should capture those metrics accurately.

P450-mediated resistance in Myzus persicae (Sulzer) (Hemiptera: Aphididae) reduces the efficacy of neonicotinoid seed treatments in Brassica napus.

BACKGROUND: The prophylactic use of seeds treated with neonicotinoid insecticides remains an important means of controlling aphid pests in canola (Brassica napus) crops in many countries. Yet, one of the most economically important aphid species worldwide, the peach potato aphid (Myzus persicae), has evolved mechanisms which confer resistance to neonicotinoids, including amplification of the cytochrome P450 gene, CYP6CY3. While CYP6CY3 amplification has been associated with low-level resistance to several neonicotinoids in laboratory acute toxicity bioassays, its impact on insecticide efficacy in the field remains unresolved. In this study, we investigated the impact of CYP6CY3 amplification on the ability of M. persicae to survive neonicotinoid exposure under laboratory and semi-field conditions. RESULTS: Three M. persicae clones, possessing different copy numbers of CYP6CY3, were shown to respond differently when exposed to the neonicotinoids, imidacloprid and thiamethoxam, in laboratory bioassays. Two clones, EastNaernup209 and Osborne171, displayed low levels of resistance (3-20-fold), which is consistent with previous studies. However, in a large-scale semi-field trial, both clones showed a surprising ability to survive and reproduce on B. napus seedlings grown from commercial rates of neonicotinoid-treated seed. In contrast, an insecticide-susceptible clone, of wild-type CYP6CY3 copy number, was unable to survive on seedlings treated in the same manner. CONCLUSION: Our findings suggest that amplification of CYP6CY3 in M. persicae clones substantially impairs the efficacy of neonicotinoid seed treatments when applied to B. napus. These findings highlight the potentially important real-world implications of resistances typically considered to be 'low level' as defined through laboratory bioassays. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

TAD Evolutionary and functional characterization reveals diversity in mammalian TAD boundary properties and function.

Topological associating domains (TADs) are self-interacting genomic units crucial for shaping gene regulation patterns. Despite their importance, the extent of their evolutionary conservation and its functional implications remain largely unknown. In this study, we generate Hi-C and ChIP-seq data and compare TAD organization across four primate and four rodent species, and characterize the genetic and epigenetic properties of TAD boundaries in correspondence to their evolutionary conservation. We find that only 14% of all human TAD boundaries are shared among all eight species (ultraconserved), while 15% are human-specific. Ultraconserved TAD boundaries have stronger insulation strength, CTCF binding, and enrichment of older retrotransposons, compared to species-specific boundaries. CRISPR-Cas9 knockouts of two ultraconserved boundaries in mouse models leads to tissue-specific gene expression changes and morphological phenotypes. Deletion of a human-specific boundary near the autism-related AUTS2 gene results in upregulation of this gene in neurons. Overall, our study provides pertinent TAD boundary evolutionary conservation annotations, and showcase the functional importance of TAD evolution.

TAD evolutionary and functional characterization reveals diversity in mammalian TAD boundary properties and function.

Topological associating domains (TADs) are self-interacting genomic units crucial for shaping gene regulation patterns. Despite their importance, the extent of their evolutionary conservation and its functional implications remain largely unknown. In this study, we generate Hi-C and ChIP-seq data and compare TAD organization across four primate and four rodent species and characterize the genetic and epigenetic properties of TAD boundaries in correspondence to their evolutionary conservation. We find 14% of all human TAD boundaries to be shared among all eight species (ultraconserved), while 15% are human-specific. Ultraconserved TAD boundaries have stronger insulation strength, CTCF binding, and enrichment of older retrotransposons compared to species-specific boundaries. CRISPR-Cas9 knockouts of an ultraconserved boundary in a mouse model lead to tissue-specific gene expression changes and morphological phenotypes. Deletion of a human-specific boundary near the autism-related AUTS2 gene results in the upregulation of this gene in neurons. Overall, our study provides pertinent TAD boundary evolutionary conservation annotations and showcases the functional importance of TAD evolution.

Virtual Ultrasound Training in a Multinational Teaching Program.

INTRODUCTION: Before the COVID-19 pandemic, a 1-week in-person Clinical Ultrasound Course was taught in African nations as part of a U.S. Department of State-funded program that supports and trains African peacekeepers serving with the United Nations and African Union. In order to maintain active engagement with host nations despite the travel restrictions due to the COVID-19 pandemic, portions of the course were taught virtually in 2021 to providers in Ghana, Senegal, and Rwanda. An abbreviated course was delivered covering the Focused Assessment with Sonography in Trauma (FAST) trauma exam and vascular access. The goal of this study is to assess the effectiveness of the Clinical Ultrasound Course while taught in a virtual classroom. MATERIALS AND METHODS: Thirty-six participants enrolled in the program. Participants completed a pre-course survey before the training. Training consisted of a pre-recorded lecture followed by hands-on ultrasound instruction. After the training, participants completed a post-course survey. Eight participants were excluded from the study because they did not complete both surveys. Survey questions assessed the participants' comfort with ultrasound mechanics and the FAST exam. Participant responses were measured utilizing a visual analog scale. RESULTS: Based on survey results, all participants gained a better understanding of the indications and limitations of the FAST exam from the virtual Clinical Ultrasound Course. All participants also felt more comfortable using ultrasound in clinical practice than they did before the course. Ghanaian and Senegalese participants showed greater improvement in all areas studied than Rwandan participants. This is likely due to the fact that the Rwandan hospital system had prior ultrasound training and a higher baseline understanding than their counterparts. The Rwandan hospital system had previously completed the in-person sessions of the Clinical Ultrasound Course and therefore had some institutional knowledge, while the Ghanaians and Senegalese took the course for the first time during this study. CONCLUSION: Virtual delivery of the Clinical Ultrasound Course was successful. Participants felt more comfortable in all aspects of ultrasound taught during the course and indicated that they were more likely to use ultrasound in clinical practice. This demonstrates that virtual ultrasound teaching is a viable option for international educational programs in the future.

Bonded by nature: Humans form equally strong and reciprocated bonds with similar raised dogs and wolves.

Introduction: To explore human-canid relationships, we tested similarly socialized and raised dogs (Canis familiaris) and wolves (Canis lupus) and their trainers in a wildlife park. The aims of our study were twofold: first, we aimed to test which factors influenced the relationships that the trainers formed with the dogs or wolves and second, we investigated if the animals reacted to the trainers in accordance with the trainers' perceptions of their relationship. Methods: To achieve these goals, we assessed the relationships using a human-animal bonds survey, which the trainers used to rate the bonds between themselves and their peers with the canids, and by observing dyadic trainer-canid social interactions. Results: Our preliminary results given the small sample size and the set-up of the research center, demonstrate that our survey was a valid way to measure these bonds since trainers seem to perceive and agree on the strength of their bonds with the animals and that of their fellow trainers. Moreover, the strength of the bond as perceived by the trainers was mainly predicted by whether or not the trainer was a hand-raiser of the specific animal, but not by whether or not the animal was a wolf or a dog. In the interaction test, we found that male animals and animals the trainers felt more bonded to, spent more time in proximity of and in contact with the trainers; there was no difference based on species. Discussion: These results support the hypothesis that wolves, similarly to dogs, can form close relationships with familiar humans when highly socialized (Canine Cooperation Hypothesis). Moreover, as in other studies, dogs showed more submissive behaviors than wolves and did so more with experienced than less experienced trainers. Our study suggests that humans and canines form differentiated bonds with each other that, if close, are independent of whether the animal is a wolf or dog.

DNA Methylation Analysis of Turner Syndrome BAV.

Turner Syndrome (TS) is a rare cytogenetic disorder caused by the complete loss or structural variation of the second sex chromosome. The most common cause of early mortality in TS results from a high incidence of left-sided congenital heart defects, including bicuspid aortic valve (BAV), which occurs in about 30% of individuals with TS. BAV is also the most common congenital heart defect in the general population with a prevalence of 0.5-2%, with males being three-times more likely to have a BAV than females. TS is associated with genome-wide hypomethylation when compared to karyotypically normal males and females. Alterations in DNA methylation in primary aortic tissue are associated with BAV in euploid individuals. Here we show significant differences in DNA methylation patterns associated with BAV in TS found in peripheral blood by comparing TS BAV (n = 12), TS TAV (n = 13), and non-syndromic BAV (n = 6). When comparing TS with BAV to TS with no heart defects we identified a differentially methylated region encompassing the BAV-associated gene MYRF, and enrichment for binding sites of two known transcription factor contributors to BAV. When comparing TS with BAV to euploid women with BAV, we found significant overlapping enrichment for ChIP-seq transcription factor targets including genes in the NOTCH1 pathway, known for involvement in the etiology of non-syndromic BAV, and other genes that are essential regulators of heart valve development. Overall, these findings suggest that altered DNA methylation affecting key aortic valve development genes contributes to the greatly increased risk for BAV in TS.