Progressive ankylosis homolog (ANKH) is a transmembrane protein essential for regulating bone mineralization through the export of nucleoside triphosphates, primarily adenosine triphosphate (ATP), and citrate into the extracellular matrix. Exported ATP is hydrolyzed by ectonucleotide pyrophosphatase/phosphodiesterase 1 into AMP and inorganic pyrophosphate (PPi). Progressive ankylosis homolog is widely expressed across tissues, where it limits ectopic mineralization of tissues and other roles. Its functional role is particularly prominent in mineralizing cells such as osteoblasts and chondrocytes that maintain the balance between mineral formation and inhibition required for healthy skeletal function. Mutations in ANKH cause 2 main mineralization disorders: craniometaphyseal dysplasia, characterized by progressive craniofacial bone thickening, and calcium pyrophosphate deposition disease (CPPD), marked by crystal deposits causing arthritis-like joint symptoms. Functionally, ANKH operates within a coordinated regulatory axis with ectonucleotide pyrophosphatase/phosphodiesterase 1 and tissue-nonspecific alkaline phosphatase (TNAP) that governs extracellular PPi homeostasis. Although TNAP-mediated PPi hydrolysis contributes negligibly to extracellular inorganic phosphate (Pi) levels, this process remains essential for clearing PPi to prevent excessive inhibition of mineral deposition, thereby preserving the Pi/PPi ratio required for physiological mineralization. Emerging evidence also implicates ANKH in citrate export, influencing bone matrix composition, and mechanical integrity. Further, ANKH expression and function are regulated by multiple signaling pathways including Wnt, tumor necrosis factor-alpha, and FGF, as well as by post-transcriptional modifications, although these regulatory mechanisms remain poorly characterized. Current mouse models, primarily knock-in lines carrying craniometaphyseal dysplasia-associated mutations, only partially recapitulate human phenotypes while comparable models for CPPD-associated mutations remain unavailable. Addressing these gaps by elucidating mutation-specific mechanisms, signaling networks, and developing improved animal models will be critical to advance targeted therapies for ANKH-related mineralization disorders and improve patient outcomes.

The vertebrate skull is composed of bones derived from neural crest cells and mesoderm. The evolutionary capacity of the skull has been linked, in part, to the emergence of neural crest cells; however, this increased capacity for evolutionary change requires that variation within neural crest- and mesoderm-derived bones remains partly autonomous. One way to assess whether tissue origin leads to discrete patterns of variation is through measures of morphological integration and modularity. In this study, we use a neural crest-specific gap junction alpha-1 (Gja1) knockout mouse model (Cx43cKO) to determine the effect of tissue origin on skull integration and modularity. Micro-computed tomography images obtained from embryonic, newborn, and 2-month Cx43cKO and wildtype (Cx43WT) mice were used to measure and compare skull shape, size, integration, and modularity between genotypes. To determine if the phenotypic differences observed between genotypes reflect Cx43 function, mRNA expression data for markers of bone differentiation were measured from the neural crest-derived frontal bones and mesoderm-derived parietal and occipital bones. We found that patterns of integration and modularity change over development and these changes correspond with differences in Cx43 expression throughout the lifespan. Most interestingly, the patterns of developmental integration and modularity we observed at birth were influenced most greatly by tissue interactions, rather than Cx43 expression in the bones. Ultimately, our findings highlight the power of experimental models for investigating integration and modularity and the importance of tissue interactions in skull development.

IntroductionChronic prenatal alcohol exposure (PAE) and severe juvenile stress independently contribute to hyperactive and depressive behavioral phenotypes, with their combination exacerbating these effects. However, while chronic PAE and traumatic juvenile stress are well-studied, little is known about the impact of early, acute PAE and mild juvenile stress on hyperactivity and depression. This knowledge gap is clinically relevant, as these milder early-life insults are common in Western societies. Here, we provide the first investigation into the effects of early, acute PAE and juvenile sub-chronic, unpredictable, mild stress (SUMS)—both independently and in combination—on hyperactivity and depressive-like behaviors in mice throughout the lifespan.MethodsWe assessed hyperactivity through movement-related measures (i.e., distance traveled, thigmotaxis, and rearing), whereas depressive-like behaviors were evaluated using the u-shaped two-choice field and forced swim tests. Behavioural testing was performed on equivalent numbers of male and female offspring and repeated at juvenile, adolescent, and adult timepoints to enable assessment of sex and age effects.ResultsNeither early, acute PAE, juvenile SUMS, nor their combination induced depressive-like behaviors at any age; findings in contrast to the more severe chronic PAE and stress insults used in previous studies. However, these milder early-life insults did result in various hyperactivity phenotypes in both the male and female offspring. For example, juvenile SUMS had the strongest impact on hyperactive behaviors across both sexes, but only the adolescent females exhibited increased emotionality-associated activity. Moreover, early, acute PAE—both alone and in combination with juvenile SUMS significantly increased movement during adolescence and adulthood exclusively in male offspring.DiscussionThus, our collective findings not only indicate that early, acute PAE and juvenile SUMS influence hyperactivity in a sex- and age-dependent manner, but also highlight that their influence on hyperactive and depressive phenotypes do not simply mirror those of the more severe early-life insults. Given the potential prevalence of early, acute alcohol exposure and juvenile stress in Western society, further research is warranted to fully understand their long-term behavioral consequences.

Craniometaphyseal Dysplasia (CMD) is a rare skeletal disorder that can result from mutations in the ANKH gene. This gene encodes progressive ankylosis (ANK), which is responsible for transporting inorganic pyrophosphate (PPi) and ATP from the intracellular to the extracellular environment, where PPi inhibits bone mineralization. When ANK is dysfunctional, as in patients with CMD, the passage of PPi to the extracellular environment is reduced, leading to excess mineralization, particularly in bones of the skull. Zebrafish may serve as a promising model to study the mechanistic basis of CMD. Here we provide a detailed analysis of the zebrafish ankh paralogs, ankha and ankhb, in terms of their phylogenic relationship with ANKH in other vertebrates as well as their spatiotemporal expression patterns during zebrafish development. We found a closer evolutionary relationship exists between the zebrafish ankhb protein and its human and other “higher” vertebrate counterparts. Furthermore, we noted distinct temporal expression patterns with ankha more prominently expressed in early development stages, and ankhb expression at larval growth stages. Whole mount in situ hybridization was used to compare spatial expression patterns of each paralog during bone development. Both paralogs showed strong expression in the craniofacial region as well as the notochord and somites, with only subtle patterning differences. Given the substantial overlap in spatiotemporal expression of ankha and ankhb , the exact roles of these genes remain speculative. However, this study lays the groundwork for functional analyses of each ankh paralog and the potential of using zebrafish to find possible targeted therapies for CMD. ### Competing Interest Statement The authors have declared no competing interest.

Craniometaphyseal Dysplasia (CMD) is a rare skeletal disorder that can result from mutations in the ANKH gene. This gene encodes progressive anksylosis (ANK), which is responsible for transporting inorganic pyrophosphate (PPi) and ATP from the intracellular to the extracellular environment, where PPi inhibits bone mineralization. When ANK is dysfunctional, as in patients with CMD, the passage of PPi to the extracellular environment is reduced, leading to excess mineralization, particularly in bones of the skull. Zebrafish may serve as a promising model to study the mechanistic basis of CMD. Here, we provide a detailed analysis of the zebrafish Ankh paralogs, Ankha and Ankhb, in terms of their phylogenic relationship with ANK in other vertebrates as well as their spatiotemporal expression patterns during zebrafish development. We found that a closer evolutionary relationship exists between the zebrafish Ankhb protein and its human and other vertebrate counterparts, and stronger promoter activity was predicted for ankhb compared to ankha. Furthermore, we noted distinct temporal expression patterns, with ankha more prominently expressed in early development stages, and both paralogs also being expressed at larval growth stages. Whole-mount in situ hybridization was used to compare the spatial expression patterns of each paralog during bone development, and both showed strong expression in the craniofacial region as well as the notochord and somites. Given the substantial overlap in spatiotemporal expression but only subtle patterning differences, the exact roles of these genes remain speculative. In silico analyses predicted that Ankha and Ankhb have the same function in transporting PPi across the membrane. Nevertheless, this study lays the groundwork for functional analyses of each ankh paralog and highlights the potential of using zebrafish to find possible targeted therapies for CMD.

In our role as medical educators and researchers, we support in the strongest manner possible the personal autonomy and self-determination of our students, patients, and colleagues. A fundamental value of the medical profession is for the patient to have personal autonomy in their healthcare decisions, including how they would like to be identified. It is, and will continue to be, an important priority to be taught and encouraged throughout healthcare education,1 and it begins in the anatomy laboratory with the proper and respectful treatment of body donors.2 Learning this value continues with respecting and accepting this same autonomy in fellow students, staff, faculty, and patients. There are some who question whether foregrounding the value of personal autonomy and the right to self-identification in health professions education is an appropriate use of resources and teaching time in an already crowded curriculum. Let us be clear: understanding this value and addressing it in the correct language3 are of paramount importance to the well-being of all involved in healthcare and should be a recurring theme throughout their education and career.4 Personal autonomy is one of the four pillars of bioethics and is a strong antidote to the paternalism and mistakes of recent eras of healthcare delivery. Any individual should be able to make their personal identity known without fear or ridicule, and this is especially important in healthcare fields. Time and resources should be devoted to educating our future healthcare professionals in the proper way to address patients, students, and colleagues. This should occur during all phases of healthcare education and should be practiced and exemplified by those who teach in healthcare. Appropriate identification of individuals, and their right to choose how they wish to be identified, is a fundamental tenet of healthcare professionalism and demands recognition as an integral principle of human rights. Complete healthcare starts with a thorough understanding of the patient, and that includes respect of personal autonomy. Acquiring skills and knowledge around the appropriate application of such principles is necessary as one of many essential components of medical education and training.

BACKGROUND:We previously determined a nonlinear relationship between connexin 43 (Cx43) function and craniofacial phenotypic variation in the mutant mouse model G60S/+, and that this variation was driven by nasal bone deviation. While nonlinearities in the genotype-phenotype map appear to be common, few studies have looked at the developmental processes that underlie this nonlinearity. Here, we investigated the potential tissue-level developmental determinants of the variation in nasal bone phenotype in G60S/+ mice through postnatal development.RESULTS:The deviated nasal bone phenotype arises by postnatal day 21 and becomes more severe by 3 months in G60S/+ mice. Measures of nasal bone remodeling including the number of osteoclasts, mineralizing surface, mineral apposition rate, and bone formation rate are significantly greater in G60S/+ mice compared to wild-type mice at 2 months, but these differences do not correspond with nasal bone deviation. The degree of nasal bone deviation does significantly and negatively correlate with the ratio between nasal bone and cartilaginous nasal septum length.CONCLUSIONS:Our findings indicate that the mean phenotypic changes observed between G60S/+ and wild-type mice are due to reduced bone growth, but the increased phenotypic variation found within mutant mice is due to discordant growth between nasal cartilage and bone.

PURPOSE:Although several different contouring instructional programs are available to radiation oncologists and trainees, very little is known about which methods and resources benefit learners most, and whether some learners may need alternate forms of instruction. This study aimed to determine the factors that were predictors of learners' success in anatomy, radiology, and contouring education. METHODS AND MATERIALS:Participants in the online and face-to-face Anatomy and Radiology Contouring (ARC) Bootcamp completed pre- and postintervention evaluations that assessed anatomy/radiology knowledge, contouring skills, self-confidence, and spatial ability. Baseline factors were assessed as predictors of outcomes across multiple educational domains. RESULTS:One hundred and eighty (face-to-face: n = 40; online: n = 140) participants enrolled in the ARC Bootcamp, and 57 (face-to-face: n = 30; online: n = 27) participants completed both evaluations. Of the participants enrolled, 37% were female, and most were radiation oncology residents (62%). In the anatomy/radiology knowledge testing, all quartiles (based on baseline performance) improved numerically; however, the largest improvements occurred in learners with the lowest baseline scores (P < .001). At the end of the Bootcamp, learners with lower-performing scores did not reach the level of learners with the highest baseline scores (Bonferroni-corrected P < .001). Regarding the contouring assessment, improvements were only evident for the participants with lower-performing baseline scores (P < .05). Spatial anatomy skills, as measured by the spatial anatomy task, were correlated to contouring ability. Overall, the greatest improvements were seen for learners in postgraduate year 1 to 3, those with no previous rotation experience in a given discipline, and those who attended from other programs (ie, medical physics residents and medical students). CONCLUSIONS:The ARC Bootcamp improved all levels of performers' anatomy and radiology knowledge but only lower-performers' contouring ability. The course alone does not help lower-performing learners reach the abilities of higher-performers. The ARC Bootcamp tends to be most beneficial for participants with less radiation oncology experience. Curriculum modifications can be made to help support ARC Bootcamp participants with lower performing scores.

Background As new treatments and technologies have been introduced in radiation oncology, the clinical roles of radiation therapists (RTs) have expanded. However, there are few formal learning opportunities for RTs. An online, anatomy, radiology and contouring bootcamp (ARC Bootcamp) originally designed for medical residents was identified as a prospective educational tool for RTs. The purpose of this study was to evaluate an RT edition of the ARC Bootcamp on knowledge, contouring, and confidence, as well as to identify areas for future modification. Methods Fifty licensed RTs were enrolled in an eight-week, multidisciplinary, online RT ARC Bootcamp. Contouring practice was available throughout the course using an online contouring platform. Outcomes were evaluated using a pre-course and post-course multiple-choice quiz (MCQ), contouring evaluation and qualitative self-efficacy and satisfaction survey. Results Of the fifty enrolled RTs, 30 completed the course, and 26 completed at least one of the post-tests. Nineteen contouring dice similarity coefficient (DSC) scores were available for paired pre- and post-course analysis. RTs demonstrated a statistically significant increase in mean DSC scoring pooled across all contouring structures (mean ± SD improvement: 0.09 ± 0.18 on a scale from 0 to 1, p=0.020). For individual contouring structures, 3/15 reached significance in contouring improvement. MCQ scores were available for 26 participants and increased after RT ARC Bootcamp participation with a mean ± SD pre-test score of 18.6 ± 4.2 (46.5%); on a 40-point scale vs. post-test score of 24.5 ± 4.3 (61.4%) (p < 0.001). RT confidence in contouring, anatomy knowledge and radiographic identification improved after course completion (p < 0.001). Feedback from RTs recommended more contouring instruction, less in-depth anatomy review and more time to complete the course. Conclusions The RT ARC Bootcamp was an effective tool for improving anatomy and radiographic knowledge among RTs. The course demonstrated improvements in contouring and overall confidence. However, only approximately half of the enrolled RTs completed the course, limiting statistical power. Future modifications will aim to increase relevance to RTs and improve completion rates.

American Journal of Biological AnthropologyEarly View MEDIA REVIEW Evolutionary cell processes in primates. Volume I: Bones, brains, and muscle; Volume II: Genes, skin, energetics, breathing, and feeding. Pitirri, Edited by M. Kathleen, and Richtsmeier, Joan T., 2022. Boca Raton, FL: CRC Press, 530 pp. ISBN 0-367-43767-1. (hardback) $160.00 Katherine E. Willmore, Corresponding Author Katherine E. Willmore kwillmo2@uwo.ca orcid.org/0000-0002-8563-3354 Department of Anatomy & Cell Biology, University of Western Ontario, London, Ontario, Canada Correspondence Katherine E. Willmore, Department of Anatomy & Cell Biology, University of Western Ontario, London, ONT, Canada. Email: kwillmo2@uwo.ca Contribution: Conceptualization (equal), Writing - original draft (equal), Writing - review & editing (equal)Search for more papers by this author Katherine E. Willmore, Corresponding Author Katherine E. Willmore kwillmo2@uwo.ca orcid.org/0000-0002-8563-3354 Department of Anatomy & Cell Biology, University of Western Ontario, London, Ontario, Canada Correspondence Katherine E. Willmore, Department of Anatomy & Cell Biology, University of Western Ontario, London, ONT, Canada. Email: kwillmo2@uwo.ca Contribution: Conceptualization (equal), Writing - original draft (equal), Writing - review & editing (equal)Search for more papers by this author First published: 01 July 2022 https://doi.org/10.1002/ajpa.24582Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Early ViewOnline Version of Record before inclusion in an issue RelatedInformation

Most features of the craniofacial complex that are necessary for feeding derive from cranial neural crest cells (cNCCs). These pluripotent stem cells originate from the dorsal neural folds and undergo a series of coordinated processes such as induction, epithelial‐to‐mesenchymal transition (EMT), migration, and differentiation. While most of these processes have been well‐studied for their roles in development, much remains to be known about the final step cNCCs must undergo, differentiation. Connexin‐43 (Cx43) is a gap junctional protein that is widely expressed, evolutionarily conserved, and has been well‐studied for its impacts on neural crest and bone. Previous research has shown that loss of Cx43 function impacts neural crest EMT and migration and can delay early osteoblast and chondrocyte differentiation; however, it is unknown how Cx43 deficiency in cNCCs impacts cNCC differentiation into osteochondrogenic lineages and the resultant impacts on craniofacial morphology. Therefore, the purpose of this project is to determine the effects of a neural crest‐specific loss of Cx43 on osteogenic and chondrogenic differentiation, and how these effects may alter skull phenotype.

BACKGROUND:Prenatal alcohol exposure (PAE) can result in developmental defects that include growth restriction, craniofacial anomalies, and cognitive behavioral deficits, though the presence and severity of these adverse outcomes can vary dramatically among exposed individuals. Preclinical animal models have demonstrated that the dose and timing of PAE account for much, but not all, of this phenotypic variation, suggesting that additional factors mitigate the effects of PAE. Here, we used a mouse model to investigate whether maternal age modulates the effects of PAE on the severity and variation in offspring growth and craniofacial outcomes.METHODS:Nulliparous C57BL/6N dams received either an intraperitoneal injection of ethanol (EtOH) or vehicle solution on gestational day 7.5. Dams were divided into four groups: (1) EtOH-treated young dams (6 to 10 weeks); (2) control young dams; (3) EtOH-treated old dams (6 to 7 months); and (4) old control dams. Neonate offspring growth restriction was measured through body mass and organ-to-body mass ratios, while skeletal craniofacial features were imaged using micro-CT and analyzed for size, shape, and variation.RESULTS:PAE and advanced maternal age each increased the risk of low birthweight and growth restriction in offspring, but these factors in combination changed the nature of the growth restriction. Similarly, both PAE and advanced maternal age individually caused changes to craniofacial morphology such as smaller skull size, dysmorphic skull shape, and greater skull shape variation and asymmetry. Interestingly, while the combination of PAE and advanced maternal age did not affect mean skull shape or size, it significantly increased the variation and asymmetry of those measures.CONCLUSION:Our results indicate that maternal age modulates the effects of PAE, but that the effects of this combination on offspring outcomes are more complex than simply scaling the effects of either factor.

Background We compared skull shape and variation among genetically modified mice that exhibit different levels of connexin43 (Cx43) channel function, to determine whether Cx43 contributes to craniofacial phenotypic robustness. Specifically, we used two heterozygous mutant mouse models (G60S/+ and I130T/+) that, when compared to their wildtype counterparts, have an ~80% and ~50% reduction in Cx43 function, respectively. Results Both mutant strains showed significant differences in skull shape compared to wildtype littermates and while these differences were more severe in the G60S/+ mouse, shape differences were localized to similar regions of the skull in both mutants. However, increased skull shape variation was observed in G60S/+ mutants only. Additionally, covariation of skull structures was disrupted in the G60S/+ mutants only, indicating that while a 50% reduction in Cx43 function is sufficient to cause a shift in mean skull shape, the threshold for Cx43 function for disrupting craniofacial phenotypic robustness is lower. Conclusions Collectively, our results indicate Cx43 can contribute to phenotypic robustness of the skull through a nonlinear relationship between Cx43 gap junctional function and phenotypic outcomes.

Connexin43 (Cx43), a gap-junctional protein, is known to play a role in bone development and remodelling. Our understanding of how Cx43 helps regulate bone remodelling has largely been gained through studies of long bones from a variety of Cx43 knockout mouse models, wherein disrupted osteoblast, osteoclast and osteocyte function cause altered bone morphology. However, clinically, altered Cx43 function manifests as the rare disorder oculodentodigital dysplasia (ODDD), caused by mutations in GJA1, the gene that encodes Cx43. Patients with ODDD display a suite of skeletal defects that are largely localized to craniofacial structures. While the important of Cx43 in long bone remodelling has been established, it is unknown if it plays a similar role in the skull. Thus here, using a mouse model of ODDD heterozygous for a mutation in Gja1 (G60S/+) and with known abnormal craniofacial morphology, we investigate the role of Cx43 in remodelling within the skull. Given that skeletally mature G60S/+ mice present with variable curvature of the nasal bones, we measure and compare phenotype and remodelling in nasal bones between G60S/+ and wildtype (WT) mice using micro-CT imaging, dynamic calcein histology, immunohistochemistry and tartrate-resistant acid phosphatase (TRAP) staining. Results from micro-CT imaging of G60S/+ and WT littermates over a post-natal developmental timeline from post-natal day 7, 14, 21 and three-months of age have shown that this craniofacial asymmetry is absent in younger mice, appears by post-natal day 21 and gets progressively worse with age. By three-months of age, one third of G60S/+ mice display the curved nasal bone phenotype whereas this phenotype is absent in WT mice, suggesting that the phenotype is caused by altered bone remodelling rather than disruption of initial development. Immunohistochemical staining for sclerostin and TRAP staining is greatly pronounced in G60S/+ nasal bones and corresponds to regions that display a concave curvature. These results suggest that the abnormal nasal bone curvature phenotype displayed in G60S/+ mice could be due to altered osteoclast activity and bone resorption and indicate that Cx43 does indeed help regulate bone remodelling in the skull. Our findings provide novel and exciting insights into the role of Cx43 in bone homeostasis and suggests that this protein serves a similar function in remodelling throughout the skeleton. Additionally, our results indicate that many of the skeletal manifestations of ODDD may be due to disrupted bone remodelling, which could underlie the immense phenotypic variation among patients with ODDD.

PurposeThe Anatomy and Radiology Contouring (ARC) Bootcamp was a face-to-face (F2F) intervention providing integrated education for radiation oncology (RO) residents and medical physicists. To increase access, we launched an online offering in 2019. We evaluated the effect of the online course on participants' knowledge acquisition, contouring skills, and self-confidence by comparing it with the F2F course.Methods and MaterialsUsing modules, the online course offers content similar to that of the F2F comparator. Participants from the 2019 F2F and the 2019-2020 online course completed pre- and postevaluations assessing anatomy and radiology knowledge, contouring skills, self-confidence, and course satisfaction.ResultsThere were 180 individuals enrolled (F2F: n = 40; online: n = 140); 57 participants (F2F: n = 30; online: n = 27) completed both evaluations. The online course had a wider geographic participation (19 countries) than F2F (4 countries). F2F had primarily RO resident participation (80%), compared with online (41%). Both cohorts demonstrated similar improvements in self-confidence pertaining to anatomy and radiology knowledge, contouring skills, and interpreting radiology images (all P < .001). Both the online (mean ± SD improvement: 6.6 ± 6.7 on a 40-point scale; P < .001) and F2F (3.7 ± 5.7; P = .002) groups showed anatomy and radiology knowledge improvement. Only the F2F group demonstrated improvement with the contouring assessment (F2F: 0.10 ± 0.17 on a 1-point Dice scale; P = .004; online: 0.07 ± 0.16; P = .076). Both cohorts perceived the course as a positive experience (F2F: 4.8 ± 0.4 on a 5-point scale; online: 4.5 ± 0.6), stated it would improve their professional practice (F2F: 4.6 ± 0.5; online: 4.2 ± 0.8), and said they would recommend it to others (F2F: 4.8 ± 0.4; online: 4.4 ± 0.6).ConclusionsThe online ARC Bootcamp demonstrated improved self-confidence, knowledge scores, and high satisfaction levels among participants. The offering had lower completion rates but was more accessible to geographic regions, provided a flexible learning experience, and allowed for ongoing education during the COVID-19 pandemic.

BACKGROUND: The Anatomy and Radiology Contouring (ARC) Bootcamp was a face-to-face (F2F) course designed to ensure radiation oncology residents were equipped with the knowledge and skillset to use radiation therapy techniques properly. The ARC Bootcamp was proven to be a useful educational intervention for improving learners' knowledge of anatomy and radiology and contouring ability. An online version of the course was created to increase accessibility to the ARC Bootcamp and provide a flexible, self-paced learning environment. This study aimed to describe the instructional design model used to create the online offering and report participants' motivation to enroll in the course and the online ARC Bootcamp's strengths and improvement areas. METHODS: The creation of the online course followed the analysis, design, development, implementation, and evaluation (ADDIE) framework. The course was structured in a linear progression of locked modules consisting of radiology and contouring lectures, anatomy labs, and integrated evaluations. RESULTS: The online course launched on the platform Teachable in November 2019, and by January 2021, 140 participants had enrolled in the course, with 27 participants completing all course components. The course had broad geographic participation with learners from 19 different countries. Of the participants enrolled, 34% were female, and most were radiation oncology residents (56%), followed by other programs (24%), such as medical physics residents or medical students. The primary motivator for participants to enroll was to improve their subject knowledge/skill (44%). The most common strength identified by participants was the course's quality (41%), and the most common improvement area was to incorporate more course content (41%). CONCLUSIONS: The creation of the online ARC Bootcamp using the ADDIE framework was feasible. The course is accessible to diverse geographic regions and programs and provides a flexible learning environment; however, the course completion rate was low. Participants' feedback regarding their experiences will inform future offerings of the online course.

Purpose: Simulation-based medical education is an effective tool for medical teaching, but simulation-based medical education deployment in radiation oncology (RO) is limited. Flexible nasopharyngoscopy (FNP), an essential skill for RO residents, requires practice that typically occurs on volunteer patients, introducing the potential for stress and discomfort. We sought to develop a high-fidelity simulator and intervention that provides RO residents the opportunity to develop FNP skills in a low-pressure environment. Methods and Materials: Computed tomography images were used to create an anatomically accurate 3-dimensional-printed model of the head and neck region. An intervention incorporating didactic instruction, multimedia content, and FNP practice on the model was designed and administered to RO residents attending the Anatomy and Radiology Contouring Bootcamp. Participants completed pre- and postintervention evaluations of the training session and model fidelity, and self-assessments of FNP skill and confidence performing FNP. Participants were video recorded performing FNP pre- and postintervention. Videos were scored by a blinded observer on a predefined rubric. Changes in scores were evaluated using the Wilcoxon signed-rank test. Results: Twenty-four participants from 17 institutions and 4 countries completed the intervention, 50% were women, and most were senior residents. Postintervention, FNP confidence and FNP performance improved significantly (mean +/- standard deviation on a 10-point scale: 1.8 +/- 1.8, P < .001; 2.2 +/- 2.0, P < .001, respectively). Participants felt the model was helpful (mean +/- standard deviation on a 5-point scale: 4.2 +/- 0.6), anatomically correct (4.1 +/- 0.9), and aided in spatial comprehension (4.3 +/- 0.8). Overall satisfaction for the intervention was high (4.3 +/- 0.8). Participants strongly agreed the intervention should be integrated into RO training programs (4.3 +/- 0.8). Conclusions: A 3-dimensional-printed model and associated intervention were effective at improving FNP performance and the teaching method was rated highly by participants. RO residents may benefit from broader dissemination of this technique to improve trainee performance. (C) 2020 Elsevier Inc. All rights reserved.

The craniofacial skeleton is an intricate structure that forms through complex cell‐cell interactions across neural crest and mesoderm derived cell populations. Proper cell‐cell communication during the migration and differentiation of cranial neural crest cells is critical for normal craniofacial development and this intercellular communication is thought to be facilitated through gap junctions – membrane channels composed of connexin proteins. In particular, connexin‐43 (Cx43) has been shown to be expressed in pre‐migratory and migratory cranial neural crest cells and plays a role in neural crest epithelial‐to‐mesenchyme transition. Additionally, within the skull, neural crest‐derived regions show the greatest morphological changes in mice wherein Cx43 has been globally ablated providing further evidence that Cx43 is important for proper neural crest function and ultimately proper development of the skull. To date, there have been no studies that have directly looked at the role of Cx43 in cranial neural crest cells in a mouse model. Thus, we have developed a neural crest‐specific Cx43 knockout mouse to address this gap in knowledge. In this study we (1) verify that Cx43 ablation is limited to neural crest cell and (2) characterize and compare craniofacial size and shape among wildtype, heterozygous and homozygous knockout mice.Heterozygous Wnt1‐Cre2 recombinase mice were bred with homozygous floxed Cx43 mice to create offspring which were bred to express the genotype, Wnt1‐Cre2/+:Cx43fl/+. These mice were bred to create: control mice (Wnt‐1Cre2/+:Cx43+/+), heterozygous conditional knock‐outs (Wnt1‐Cre2/+:Cx43fl/fl) and homozygous conditional knock‐outs (Wnt1‐Cre2+/+:Cx43fl/fl). To verify the knockout, paraffin embedded sagittal sections of embryonic and post‐natal mouse hearts and skulls were tagged with Wnt1 and Cx43 antibodies. RT‐PCR, and Western blotting were performed on neural crest and mesoderm derived newborn skull bones, to verify the expression levels of Cx43 mRNA and protein respectively. Newborn skulls from each genotype were imaged using high resolution micro‐CT and these images were used to quantify and compare skull morphology among genotypes using statistical analyses of shape.We have confirmed that Cx43 ablation is limited to neural crest cells in our mice. Analyses of skull shape comparisons among genotypes is ongoing with our initial findings indicating that mice homozygous for the knockout demonstrate overt changes to skull morphology, heterozygous mouse skulls are qualitatively similar to control mouse skulls. We predict that statistical analyses will uncover that only the bones derived from neural crest cells will display morphological differences among genotypes.Our study addresses a fundamental gap of the role connexin‐43 plays in cranial neural crest function and subsequent development of much of the skull.Support or Funding InformationFunding: Natural Sciences and Engineering Research Council of Canada (NSERC) Discovery Grant (R5211A02)

Prenatal alcohol exposure (PAE) can result in fetal alcohol spectrum disorder (FASD), defined as a continuum of developmental defects including growth restriction, cognitive‐behavioral deficits, and craniofacial anomalies. Despite the known consequences of PAE, there is great variation in craniofacial and behavioral outcomes of affected children even when considering timing and quantity of exposure; phenotypic variation which could arise from maternal differences. In the present study, we asked: Does maternal age influence the nature and extent of growth restriction and craniofacial defects of PAE in offspring? To address this question, we used a C57BL/6 mouse model, as it enables precise control of dosing, gestational timing and maternal age. Dams were divided into four groups: 1. ethanol‐treated young dams (6–10 weeks); 2. control young dams; 3. ethanol‐treated old dams (6–7 months); and 4. control old dams. All dams were injected with either ethanol (treatment groups) or vehicle solution (control groups) on gestational day 7.5. Offspring growth restriction was analyzed by measures of body mass, liver mass, and body‐to‐liver mass ratio at birth. Micro‐CT scans of newborn soft tissue and skeletal craniofacial features underwent 3D analyses of shape and size to compare craniofacial phenotypes among treatment groups. Initial results of young ethanol‐treated and control groups demonstrated that ethanol‐treated offspring were born prematurely and showed significant growth restriction. Both soft tissue and skeletal craniofacial features were altered in the ethanol‐treated group, consistent with the FASD phenotype. Surprisingly, craniofacial shape of ethanol‐treated offspring was less variable than controls. Data collection for the old dam groups is ongoing, and we predict that offspring of old dams will have more severe growth and phenotypic consequences from PAE, exhibiting greater phenotypic variation compared to offspring of young dams. As many women are delaying childbirth, there is an increasing need to carryout preclinical studies with appropriate experimental controls to better understand the relative risk of PAE and maternal age, and ultimately contribute to public health policy for pregnant women.Support or Funding InformationFunded by Children’s Health Research Institute, Internal Research Grant Fund to KEW “The Effects of Moderate Prenatal Alcohol Exposure on Craniofacial Morphology and Development” R5211A04