Structural analysis of the female reptile reproductive system by micro-computed tomography and optical coherence tomography†.

Volumetric data provide unprecedented structural insight to the reproductive tract and add vital anatomical context to the relationships between organs. The morphology of the female reproductive tract in non-avian reptiles varies between species, corresponding to a broad range of reproductive modes and providing valuable insight to comparative investigations of reproductive anatomy. However, reproductive studies in reptilian models, such as the brown anole studied here, have historically relied on histological methods to understand the anatomy. While these methods are highly effective for characterizing the cell types present in each organ, histological methods lose the 3D relationships between images and leave the architecture of the organ system poorly understood. We present the first comprehensive volumetric analyses of the female brown anole reproductive tract using two non-invasive, non-destructive imaging modalities: micro-computed tomography (microCT) and optical coherence tomography (OCT). Both are specialized imaging technologies that facilitate high-throughput imaging and preserve three-dimensional information. This study represents the first time that microCT has been used to study all reproductive organs in this species and the very first time that OCT has been applied to this species. We show how the non-destructive volumetric imaging provided by each modality reveals anatomical context including orientation and relationships between reproductive organs of the anole lizard. In addition to broad patterns of morphology, both imaging modalities provide the high resolution necessary to capture details and key anatomical features of each organ. We demonstrate that classic histological features can be appreciated within whole-organ architecture in volumetric imaging using microCT and OCT, providing the complementary information necessary to understand the relationships between tissues and organs in the reproductive system. This side-by-side imaging analysis using microCT and OCT allows us to evaluate the specific advantages and limitations of these two methods for the female reptile reproductive system.

Cystic ovary disease (COD) alters structure and function of the bovine oviduct.

Cystic ovary disease (COD) is a common cause of subfertility in dairy cattle. Therefore, the aim of this study was to provide novel concepts for cyst classification and to investigate the effects of COD on tubal microarchitecture, oviductal metabolic function, and the formation of the sperm reservoir. Bovine Fallopian tubes affected by follicular cysts, follicular cysts with luteinization and luteal cysts were investigated by a variety of microscopic and histological techniques and compared to control cows in metestrus and diestrus. We defined three types of cysts involved in COD, each of which had a characteristic wall thickness, inner wall appearance and cellular pattern within the cyst aspirate. Regarding the Fallopian tube, each cyst type was associated with a characteristic morphology, specifically the microarchitecture of the folds in ampulla, epithelial cell ratios, and ciliated/secretory cell size and form. Furthermore, each cyst type showed different patterns of tubal glycoprotein and acidic mucopolysaccharide synthesis, which was highly variable as compared to the controls. Our studies are the first to characterize the effects of COD on the Fallopian tube, which promotes the establishment of novel, cyst-specific therapeutic concepts in cattle and helps gain a holistic view of the causes of subfertility in cows with COD.

An audit of antibiotic prescribing in primary care 2019-2020 in Dunmanway Primary Care Centre.

INTRODUCTION: Antibiotics are often the most common medication prescribed by general practitioners (GPs) and are often expected by patients despite campaigns such as Under the Weather. Antibiotic resistance is increasing in the community. The Health Service Executive (HSE) has issued 'Guidelines for Antimicrobial Prescribing in Primary Care in Ireland' aiming to optimise safe prescribing. This audit aims to analyse change in quality of prescribing after educational intervention. METHODS: GP prescribing patterns were analysed over a week in October 2019 and re-audited in February 2020. Anonymous questionnaires detailed demographics, condition and antibiotic details. Educational intervention included texts, information and review of current guidelines. Data were analysed on a password protected spreadsheet. The HSE Guidelines for Antimicrobial Prescribing in Primary Care were taken as reference standard. A standard of 90% compliance for choice of antibiotic and 70% compliance for dose and course was agreed. RESULTS: FindingsAuditRe-AuditNumber prescriptions4024Number delayed scripts4/40=10%1/24=4.2%Adult37/40=92.5%19/24=79.2%Child3/40=7.5%5/24=20.8%IndicationURTI22.50%25%LRTI10%4%Other RTI37.50%42%UTI20%29%Skin12.50%0%Gynaecological2.50%0%2+ Infections 5%0%Co-amoxiclav17.50%12.50%AdherenceChoice37/40=92.5%22/24=91.7%Dose28/39=71.8%17/24=70.8%Course28/40=70%12/24=50%Discussion: Excellent antibiotic choice and dose concordance with guidelines was noted, with both phases meeting the set standards. Suboptimal course compliance with guidelines occurred in the re-audit. Potential causes include concerns regarding resistance and patient factors not included. This audit included unequal number of prescriptions in each phase but are still of significance and addresses a clinically relevant topic.

Cystic ovary disease impairs transport speed, smooth muscle contraction, and epithelial ion transport in the bovine oviduct.

Cystic ovary disease (COD) is a common cause of bovine infertility but the impact of this disease on the oviduct is unknown. The aim of this study was to analyze the effects of COD on particle transport speed (PTS), ciliary beat frequency, myosalpinx contraction, and epithelial ion transport. Oviducts were obtained from cows affected by COD and compared with those of healthy, mid-diestrus cows. PTS and CBF were examined using live-cell imaging. Smooth muscle contraction and epithelial ion transport were investigated using organ baths and Ussing chambers. Our results showed that muscarinic receptors are involved in cholinergic signaling in the oviduct and that forskolin-induced cyclic AMP production is involved in active ion transport in the oviductal epithelium. Oviducts from cows with luteal cysts revealed significantly decreased PTS (p = 0.02). Further to that, in the oviducts of COD cows, the cholinergic regulation of smooth muscle contractions and active epithelial ion transport were significantly diminished (p < 0.0001). These results imply that in COD cows, oviductal transport is compromised by decreased fluid flow speed and reduced cholinergic regulation of smooth muscle contraction and ion transport. This knowledge contributes to a more comprehensive understanding of COD supporting the development of novel therapeutic concepts for infertility treatment.

Emerging from emergency pandemic pedagogy: A survey of anatomical educators in the United Kingdom and Ireland.

It is critical that academic opinion of pandemic pedagogy is comprehensively quantified in order to inform future practices. Thus, this study examines how anatomists in the United Kingdom (UK) and Republic of Ireland (ROI) perceive the teaching adaptations made in response to COVID-19, and how these adaptations have impacted their experiences teaching, their online work environment and community. Data was collected via a questionnaire from 24 anatomists across 15 universities in the UK (11) and ROI (4). With regards to teaching, 95.6% of academics have upskilled in new technologies to meet the demands of distance teaching. Academics (95.8%) preferred face-to-face delivery of practical sessions. Most universities (80.0%) reported that practical sessions will continue in a new form that ensures social distancing. However, 50.0% of academics are uncertain if these adaptations will improve student learning. Many anatomists believe that the new adaptations may hinder student-student (66.7%) and student-tutor (45.8%) interactions. Regarding assessment, 52.6% of academics preferred traditional methods to online. Remote online assessment was difficult to protect against collusion, but provided time saving opportunities for academics. Finally, in terms of working environment, 83.3% of academics stated that their workload increased; 54.2% preferred working on site rather than remotely and 79.2% think that staff interactions are better when working on site. These results demonstrate a widespread concern amongst anatomists regarding the pandemic-induced adaptations to teaching, assessment and working environment. However, important opportunities were also identified that could ultimately serve to benefit students and educators alike.

Hypoxia promotes production of neural crest cells in the embryonic head.

Hypoxia is encountered in either pathological or physiological conditions, the latter of which is seen in amniote embryos prior to the commencement of a functional blood circulation. During the hypoxic stage, a large number of neural crest cells arise from the head neural tube by epithelial-to-mesenchymal transition (EMT). As EMT-like cancer dissemination can be promoted by hypoxia, we investigated whether hypoxia contributes to embryonic EMT. Using chick embryos, we show that the hypoxic cellular response, mediated by hypoxia-inducible factor (HIF)-1α, is required to produce a sufficient number of neural crest cells. Among the genes that are involved in neural crest cell development, some genes are more sensitive to hypoxia than others, demonstrating that the effect of hypoxia is gene specific. Once blood circulation becomes fully functional, the embryonic head no longer produces neural crest cells in vivo, despite the capability to do so in a hypoxia-mimicking condition in vitro, suggesting that the oxygen supply helps to stop emigration of neural crest cells in the head. These results highlight the importance of hypoxia in normal embryonic development.

In vivo volumetric depth-resolved imaging of cilia metachronal waves using dynamic optical coherence tomography.

Motile cilia are dynamic hair-like structures covering epithelial surfaces in multiple organs. The periodic coordinated beating of cilia creates waves propagating along the surface, known as the metachronal waves, which transport fluids and mucus along the epithelium. Motile ciliopathies result from disrupted coordinated cilia beating and are associated with serious clinical complications, including reproductive disorders. Despite the recognized clinical significance, research of cilia dynamics is extremely limited. Here, we present quantitative imaging of cilia metachronal waves volumetrically through tissue layers using dynamic optical coherence tomography (OCT). Our method relies on spatiotemporal mapping of the phase of intensity fluctuations in OCT images caused by the ciliary beating. We validated our new method ex vivo and implemented it in vivo to visualize cilia metachronal wave propagation within the mouse fallopian tube. This method can be extended to the assessment of physiological cilia function and ciliary dyskinesias in various organ systems, contributing to better management of pathologies associated with motile ciliopathies.

Mouse embryo phenotyping with optical coherence tomography.

With the explosion of gene editing tools in recent years, there has been a much greater demand for mouse embryo phenotyping, and traditional methods such as histology and histochemistry experienced a methodological renaissance as they became the principal tools for phenotyping. However, it is important to explore alternative phenotyping options to maximize time and resources and implement volumetric structural analysis for enhanced investigation of phenotypes. Cardiovascular phenotyping, in particular, is important to perform in vivo due to the dramatic structural and functional changes that occur in heart development over relatively short periods of time. Optical coherence tomography (OCT) is one of the most exciting advanced imaging techniques emerging within the field of developmental biology, and this review provides a summary of how it is currently being implemented in mouse embryo investigations and phenotyping. This review aims to provide an understanding of the approaches used in optical coherence tomography and how they can be applied in embryology and developmental biology, with the overall aim of bridging the gap between biology and technology.

Strength, Weakness, Opportunity, Threat (SWOT) Analysis of the Adaptations to Anatomical Education in the United Kingdom and Republic of Ireland in Response to the Covid-19 Pandemic.

The Covid-19 pandemic has driven the fastest changes to higher education across the globe, necessitated by social distancing measures preventing face-to-face teaching. This has led to an almost immediate switch to distance learning by higher education institutions. Anatomy faces some unique challenges. Intrinsically, anatomy is a three-dimensional subject that requires a sound understanding of the relationships between structures, often achieved by the study of human cadaveric material, models, and virtual resources. This study sought to identify the approaches taken in the United Kingdom and Republic of Ireland to deliver anatomical education through online means. Data were collected from 14 different universities in the United Kingdom and Republic of Ireland and compared adopting a thematic analysis approach. Once themes were generated, they were collectively brought together using a strength, weakness, opportunity, threat (SWOT) analysis. Key themes included the opportunity to develop new online resources and the chance to engage in new academic collaborations. Academics frequently mentioned the challenge that time constrains could place on the quality and effectiveness of these resources; especially as in many cases the aim of these resources was to compensate for a lack of exposure to cadaveric exposure. Comparisons of the actions taken by multiple higher education institutions reveal the ways that academics have tried to balance this demand. Discussions will facilitate decisions being made by higher education institutions regarding adapting the curriculum and assessment methods in anatomy.

Automatic Clinical Procedure Detection for Emergency Services.

Understanding a patient's state is critical to providing optimal care. However, information loss occurs during patient hand-offs (e.g., emergency services (EMS) transferring patient care to a receiving hospital), which hinders care quality. Augmenting the information flow from an EMS vehicle to a receiving hospital may reduce information loss and improve patient outcomes. Such augmentation requires a noninvasive system that can automatically recognize clinical procedures being performed and send near real-time information to a receiving hospital. An automatic clinical procedure detection system that uses wearable sensors, video, and machine-learning to recognize clinical procedures within a controlled environment is presented. The system demonstrated how contextual information and a majority vote method can substantially improve procedure recognition accuracy. Future work concerning computer vision techniques and deep learning are discussed.

Feasibility Assessment of a Pre-Hospital Automated Sensing Clinical Documentation System.

Clinical documentation in the pre-hospital setting is challenged by limited resources and fast-paced, high-acuity. Military and civilian medics are responsible for performing procedures and treatments to stabilize the patient, while transporting the injured to a trauma facility. Upon arrival, medics typically give a verbal report from memory or informal source of documentation such as a glove or piece of tape. The development of an automated documentation system would increase the accuracy and amount of information that is relayed to the receiving physicians. This paper discusses the 12-week deployment of an Automated Sensing Clinical Documentation (ASCD) system among the Nashville Fire Department EMS paramedics. The paper examines the data collection methods, operational challenges, and perceptions surrounding real-life deployment of the system. Our preliminary results suggest that the ASCD system is feasible for use in the pre-hospital setting, and it revealed several barriers and their solutions.

High-throughput high-volume nuclear imaging for preclinical in vivo compound screening§.

BACKGROUND: Preclinical single-photon emission computed tomography (SPECT)/CT imaging studies are hampered by low throughput, hence are found typically within small volume feasibility studies. Here, imaging and image analysis procedures are presented that allow profiling of a large volume of radiolabelled compounds within a reasonably short total study time. Particular emphasis was put on quality control (QC) and on fast and unbiased image analysis. METHODS: 2-3 His-tagged proteins were simultaneously radiolabelled by 99mTc-tricarbonyl methodology and injected intravenously (20 nmol/kg; 100 MBq; n = 3) into patient-derived xenograft (PDX) mouse models. Whole-body SPECT/CT images of 3 mice simultaneously were acquired 1, 4, and 24 h post-injection, extended to 48 h and/or by 0-2 h dynamic SPECT for pre-selected compounds. Organ uptake was quantified by automated multi-atlas and manual segmentations. Data were plotted automatically, quality controlled and stored on a collaborative image management platform. Ex vivo uptake data were collected semi-automatically and analysis performed as for imaging data. RESULTS: >500 single animal SPECT images were acquired for 25 proteins over 5 weeks, eventually generating >3500 ROI and >1000 items of tissue data. SPECT/CT images clearly visualized uptake in tumour and other tissues even at 48 h post-injection. Intersubject uptake variability was typically 13% (coefficient of variation, COV). Imaging results correlated well with ex vivo data. CONCLUSIONS: The large data set of tumour, background and systemic uptake/clearance data from 75 mice for 25 compounds allows identification of compounds of interest. The number of animals required was reduced considerably by longitudinal imaging compared to dissection experiments. All experimental work and analyses were accomplished within 3 months expected to be compatible with drug development programmes. QC along all workflow steps, blinding of the imaging contract research organization to compound properties and automation provide confidence in the data set. Additional ex vivo data were useful as a control but could be omitted from future studies in the same centre. For even larger compound libraries, radiolabelling could be expedited and the number of imaging time points adapted to increase weekly throughput. Multi-atlas segmentation could be expanded via SPECT/MRI; however, this would require an MRI-compatible mouse hotel. Finally, analysis of nuclear images of radiopharmaceuticals in clinical trials may benefit from the automated analysis procedures developed.