Characterisation and correction of polarisation effects in fluorescently labelled fibres.

Many biological structures take the form of fibres and filaments, and quantitative analysis of fibre organisation is important for understanding their functions in both normal physiological conditions and disease. In order to visualise these structures, fibres can be fluorescently labelled and imaged, with specialised image analysis methods available for quantifying the degree and strength of fibre alignment. Here we show that fluorescently labelled fibres can display polarised emission, with the strength of this effect varying depending on structure and fluorophore identity. This can bias automated analysis of fibre alignment and mask the true underlying structural organisation. We present a method for quantifying and correcting these polarisation effects without requiring polarisation-resolved microscopy and demonstrate its efficacy when applied to images of fluorescently labelled collagen gels, allowing for more reliable characterisation of fibre microarchitecture.

'Misdiagnosed and Misunderstood': Insights into Rarer Forms of Dementia through a Stepwise Approach to Co-Constructed Research Poetry.

This study investigated co-constructed research poetry as a way to understand the lived experiences of people affected by rarer dementia and as a means to use poetry to convey those experiences to healthcare professionals. Using mixed methods, 71 people living with rarer dementia and care-partners (stakeholders) contributed to co-constructing 27 poems with professional poets; stakeholders' verbatim words were analysed with descriptive qualitative analysis. Stakeholders were also surveyed and interviewed about their participation. Healthcare professionals (n = 93) were surveyed to elicit their responses to learning through poetry and its acceptability as a learning tool. Poems conveyed a shared narrative of different aspects of lived experience, often owing to atypical symptoms, misunderstandings by professionals, lack of support pathways, and a continuous struggle to adapt. Stakeholder surveys indicated it was a valuable experience to both co-create and respond to the poems, whilst group interviews revealed people's experiences of the research poetry were characterised by reflection on lived experience, curiosity and exploration. Healthcare professionals' responses reinforced poetry's capacity to stimulate cognitive and affective learning specific to rare dementia support and prompt both empathy and critical thinking in practice. As the largest poetry-based study that we are aware of, this novel accessible approach of creating group poems yielded substantial information about the experiences and needs of those affected by rarer dementia and how poetry can contribute to healthcare education and training.

Urban rats are the 'fall-guy': Resident motivations for municipal rat complaints.

Rats are an important issue in cities globally. Despite their ubiquity, perceptions and concerns about rats vary with circumstance and the context in which a person interacts with them. Municipal rat management programs are a service to communities and therefore must be responsive to the varied concerns of their residents. Understanding why communities are concerned about rats can help inform rat management programs to meet the specific needs of their residents. The objective of this study was to identify why the residents of Vancouver, Canada care about rats and what they want done to address them. To do this, we qualitatively analyzed 6,158 resident complaints about rats made to the city's municipal government between January 2014 and May 2020. Using a qualitative descriptive coding process, we found that rats were a priority in a minority of cases. In general, people were more concerned about broader community issues, such as neighborhood disorder, of which rats were one part. Complaints tended to be made when problems were highly visible, nearby, and when the complainant wanted the city to take action to alleviate this issue, particularly when they were in and around their living spaces. The rates of complaints were highest in the most economically and socially deprived neighborhoods and lowest in the most privileged neighbourhoods. We synthesize this information with a view towards understanding how to develop objectives and actions for municipal management strategies that are grounded in community concerns.

Imaging actin organisation and dynamics in 3D.

The actin cytoskeleton plays a critical role in cell architecture and the control of fundamental processes including cell division, migration and survival. The dynamics and organisation of F-actin have been widely studied in a breadth of cell types on classical two-dimensional (2D) surfaces. Recent advances in optical microscopy have enabled interrogation of these cytoskeletal networks in cells within three-dimensional (3D) scaffolds, tissues and in vivo. Emerging studies indicate that the dimensionality experienced by cells has a profound impact on the structure and function of the cytoskeleton, with cells in 3D environments exhibiting cytoskeletal arrangements that differ to cells in 2D environments. However, the addition of a third (and fourth, with time) dimension leads to challenges in sample preparation, imaging and analysis, necessitating additional considerations to achieve the required signal-to-noise ratio and spatial and temporal resolution. Here, we summarise the current tools for imaging actin in a 3D context and highlight examples of the importance of this in understanding cytoskeletal biology and the challenges and opportunities in this domain.

Sox9 Accelerates Vascular Aging by Regulating Extracellular Matrix Composition and Stiffness.

BACKGROUND: Vascular calcification and increased extracellular matrix (ECM) stiffness are hallmarks of vascular aging. Sox9 (SRY-box transcription factor 9) has been implicated in vascular smooth muscle cell (VSMC) osteo/chondrogenic conversion; however, its relationship with aging and calcification has not been studied. METHODS: Immunohistochemistry was performed on human aortic samples from young and aged patients. Young and senescent primary human VSMCs were induced to produce ECM, and Sox9 expression was manipulated using adenoviral overexpression and depletion. ECM properties were characterized using atomic force microscopy and proteomics, and VSMC phenotype on hydrogels and the ECM were examined using confocal microscopy. RESULTS: In vivo, Sox9 was not spatially associated with vascular calcification but correlated with the senescence marker p16 (cyclin-dependent kinase inhibitor 2A). In vitro Sox9 showed mechanosensitive responses with increased expression and nuclear translocation in senescent cells and on stiff matrices. Sox9 was found to regulate ECM stiffness and organization by orchestrating changes in collagen (Col) expression and reducing VSMC contractility, leading to the formation of an ECM that mirrored that of senescent cells. These ECM changes promoted phenotypic modulation of VSMCs, whereby senescent cells plated on ECM synthesized from cells depleted of Sox9 returned to a proliferative state, while proliferating cells on a matrix produced by Sox9 expressing cells showed reduced proliferation and increased DNA damage, reiterating features of senescent cells. LH3 (procollagen-lysine, 2-oxoglutarate 5-dioxygenase 3) was identified as an Sox9 target and key regulator of ECM stiffness. LH3 is packaged into extracellular vesicles and Sox9 promotes extracellular vesicle secretion, leading to increased LH3 deposition within the ECM. CONCLUSIONS: These findings highlight the crucial role of ECM structure and composition in regulating VSMC phenotype. We identify a positive feedback cycle, whereby cellular senescence and increased ECM stiffening promote Sox9 expression, which, in turn, drives further ECM modifications to further accelerate stiffening and senescence.

Extracellular matrix stiffness activates mechanosensitive signals but limits breast cancer cell spheroid proliferation and invasion.

Breast cancer is characterized by physical changes that occur in the tumor microenvironment throughout growth and metastasis of tumors. Extracellular matrix stiffness increases as tumors develop and spread, with stiffer environments thought to correlate with poorer disease prognosis. Changes in extracellular stiffness and other physical characteristics are sensed by integrins which integrate these extracellular cues to intracellular signaling, resulting in modulation of proliferation and invasion. However, the co-ordination of mechano-sensitive signaling with functional changes to groups of tumor cells within 3-dimensional environments remains poorly understood. Here we provide evidence that increasing the stiffness of collagen scaffolds results in increased activation of ERK1/2 and YAP in human breast cancer cell spheroids. We also show that ERK1/2 acts upstream of YAP activation in this context. We further demonstrate that YAP, matrix metalloproteinases and actomyosin contractility are required for collagen remodeling, proliferation and invasion in lower stiffness scaffolds. However, the increased activation of these proteins in higher stiffness 3-dimensional collagen gels is correlated with reduced proliferation and reduced invasion of cancer cell spheroids. Our data collectively provide evidence that higher stiffness 3-dimensional environments induce mechano-signaling but contrary to evidence from 2-dimensional studies, this is not sufficient to promote pro-tumorigenic effects in breast cancer cell spheroids.

Obstetrics and gynecology clerkship directors' experiences advising residency applicants.

BACKGROUND: The evolving landscape of application processes for obstetrics and gynecology residency applicants poses many challenges for applicants and advisors. The lack of data coordination among national groups creates crucial gaps in information for stakeholder groups. OBJECTIVE: This study aimed to identify the current state of the advising milieu for obstetrics and gynecology residency applicants and their career advisors, the annual Association of Professors of Gynecology and Obstetrics survey focused on US clerkship directors' experiences advising students through these processes. STUDY DESIGN: A 23-item anonymous survey was developed that asked respondents about demographics and outcomes for the students that they advised through the 2021 application process and their experiences with dual applicants and students not matching. The survey was sent electronically to all obstetrics and gynecology clerkship directors with active Association of Professors of Gynecology and Obstetrics memberships in April 2021. RESULTS: Of 224 total clerkship directors, 143 (63.8%) responded to the survey, Of the 143 respondents, almost all (136 [95.1%]) served as career advisors, and 50 (35.0%) were aware of students dual applying. Furthermore, obstetrics and gynecology was rarely the backup to a more competitive specialty. For the 2021 application cycle, 79 of 143 respondents (55.2%) reported having students not successfully match into obstetrics and gynecology, with "academic concerns" followed by "poor communication skills" as the primary reasons cited for students not matching. CONCLUSION: This snapshot of clerkship directors' experiences advising students in the residency application process reveals notably high rates of dual applicants and students not matching into obstetrics and gynecology. This work fills key gaps in our knowledge of current processes and highlights the importance of career advising at multiple points during the application process.

Clerkship director confidence in medical student career advising in obstetrics and gynecology.

BACKGROUND: Given the increasing complexities of the residency application processes, there is an ever-increasing need for faculty to serve in the role of fourth-year medical student career advisors. OBJECTIVE: This study aimed to investigate obstetrics and gynecology clerkship directors' confidence and fulfillment with serving in the role of faculty career advisors. STUDY DESIGN: A 25-item electronic survey was developed and distributed to the 225 US obstetrics and gynecology clerkship directors in university-based and community-based medical schools with active memberships in the Association of Professors of Gynecology and Obstetrics. Items queried respondents on demographics, confidence in fourth-year advising, satisfaction with this aspect of their career, and resources used for advising. RESULTS: Of 225 clerkship directors, 143 (63.6%) responded to the survey. Nearly all clerkship directors (136/143 [95%]) reported advising fourth-year students. A median of 5.0 hours (interquartile range, 3.0-10.0) was spent per student in this advisory role, with 29 of 141 clerkship directors (20.5%) reporting some form of compensation for advising. Confidence in the ability to advise fourth-year medical students correlated significantly with number of years as a faculty, number of years as a clerkship director, and a higher full-time equivalent allotted as clerkship director. Fulfillment as a faculty career advisor was correlated with number of years as a clerkship director and a higher number of students advised. CONCLUSION: Obstetrics and gynecology clerkship directors regularly serve in the crucial role of faculty career advisor. Confidence in advising fourth-year students, advising fulfillment, and satisfaction with advising resources were all significantly correlated. We recommend that clerkship directors review resources available for advising and that they be provided academic time to serve as career advisors.

Extracellular matrix assembly stress initiates Drosophila central nervous system morphogenesis.

Forces controlling tissue morphogenesis are attributed to cellular-driven activities, and any role for extracellular matrix (ECM) is assumed to be passive. However, all polymer networks, including ECM, can develop autonomous stresses during their assembly. Here, we examine the morphogenetic function of an ECM before reaching homeostatic equilibrium by analyzing de novo ECM assembly during Drosophila ventral nerve cord (VNC) condensation. Asymmetric VNC shortening and a rapid decrease in surface area correlate with the exponential assembly of collagen IV (Col4) surrounding the tissue. Concomitantly, a transient developmentally induced Col4 gradient leads to coherent long-range flow of ECM, which equilibrates the Col4 network. Finite element analysis and perturbation of Col4 network formation through the generation of dominant Col4 mutations that affect assembly reveal that VNC morphodynamics is partially driven by a sudden increase in ECM-driven surface tension. These data suggest that ECM assembly stress and associated network instabilities can actively participate in tissue morphogenesis.

A method for reproducible high-resolution imaging of 3D cancer cell spheroids.

Multicellular tumour cell spheroids embedded within three-dimensional (3D) hydrogels or extracellular matrices (ECM) are widely used as models to study cancer growth and invasion. Standard methods to embed spheroids in 3D matrices result in random placement in space which limits the use of inverted fluorescence microscopy techniques, and thus the resolution that can be achieved to image molecular detail within the intact spheroid. Here, we leverage UV photolithography to microfabricate PDMS (polydimethylsiloxane) stamps that allow for generation of high-content, reproducible well-like structures in multiple different imaging chambers. Addition of multicellular tumour spheroids into stamped collagen structures allows for precise positioning of spheroids in 3D space for reproducible high-/super-resolution imaging. Embedded spheroids can be imaged live or fixed and are amenable to immunostaining, allowing for greater flexibility of experimental approaches. We describe the use of these spheroid imaging chambers to analyse cell invasion, cell-ECM interaction, ECM alignment, force-dependent intracellular protein dynamics and extension of fine actin-based protrusions with a variety of commonly used inverted microscope platforms. This method enables reproducible, high-/super-resolution live imaging of multiple tumour spheroids, that can be potentially extended to visualise organoids and other more complex 3D in vitro systems.

The time is now: addressing implicit bias in obstetrics and gynecology education.

Obstetrician-gynecologists can improve the learning environment and patient care by addressing implicit bias. Accumulating evidence demonstrates that racial and gender-based discrimination is woven into medical education, formal curricula, patient-provider-trainee interactions in the clinical workspace, and all aspects of learner assessment. Implicit bias negatively affects learners in every space. Strategies to address implicit bias at the individual, interpersonal, institutional, and structural level to improve the well-being of learners and patients are needed. The authors review an approach to addressing implicit bias in obstetrics and gynecology education, which includes: (1) curricular design using an educational framework of antiracism and social justice theories, (2) bias awareness and management pedagogy throughout the curriculum, (3) elimination of stereotypical patient descriptions from syllabi and examination questions, and (4) critical review of epidemiology and evidence-based medicine for underlying assumptions based on discriminatory practices or structural racism that unintentionally reinforce stereotypes and bias. The movement toward competency-based medical education and holistic evaluations may result in decreased bias in learner assessment. Educators may wish to monitor grades and narratives for bias as a form of continuous educational equity improvement. Given that practicing physicians may have little training in this area, faculty development efforts in bias awareness and mitigation strategies may have significant impact on learner well-being.

Nuclear fascin regulates cancer cell survival.

Fascin is an important regulator of F-actin bundling leading to enhanced filopodia assembly. Fascin is also overexpressed in most solid tumours where it supports invasion through control of F-actin structures at the periphery and nuclear envelope. Recently, fascin has been identified in the nucleus of a broad range of cell types but the contributions of nuclear fascin to cancer cell behaviour remain unknown. Here, we demonstrate that fascin bundles F-actin within the nucleus to support chromatin organisation and efficient DDR. Fascin associates directly with phosphorylated Histone H3 leading to regulated levels of nuclear fascin to support these phenotypes. Forcing nuclear fascin accumulation through the expression of nuclear-targeted fascin-specific nanobodies or inhibition of Histone H3 kinases results in enhanced and sustained nuclear F-actin bundling leading to reduced invasion, viability, and nuclear fascin-specific/driven apoptosis. These findings represent an additional important route through which fascin can support tumourigenesis and provide insight into potential pathways for targeted fascin-dependent cancer cell killing.

'We just don't have this in us ': Understanding factors behind low levels of physical activity in South Asian immigrants in Metro-Vancouver, Canada.

BACKGROUND: South Asian immigrants in western countries are at a high risk for metabolic syndrome and associated chronic disease. While a physically active lifestyle is crucial in decreasing this risk, physical activity (PA) levels among this group remain low. The objectives of this study were to explore social and cultural factors that influence PA behavior, investigate how immigration process intersects with PA behaviors to influence PA levels and to engage community in a discussion about what can be done to increase PA in the South Asian community. METHODS: For this qualitative study, we conducted four Focus Group Discussions (FGDs) among a subset of participants who were part of a larger study. FGD data was coded and analysed using directed content analysis to identify key categories. RESULTS: Participants expressed a range of opinions, attitudes and beliefs about PA. Most believed they were sufficiently active. Women talked about restrictive social and cultural norms that discouraged uptake of exercise. Post-immigration levels of PA were low due to change in type of work and added responsibilities. CONCLUSION: Health promoters need to consider social, cultural, and structural contexts when exploring possible behavior change interventions for South Asian immigrants.

Enhancing interprofessional collaboration and interprofessional education in women's health.

This article is from the 'To The Point' series from the Association of Professors of Gynecology and Obstetrics Undergraduate Medical Education Committee. The purpose of this review is to provide an understanding of the differing yet complementary nature of interprofessional collaboration and interprofessional education as well as their importance to the specialty of Obstetrics and Gynecology. We provide a historical perspective of how interprofessional collaboration and interprofessional education have become key aspects of clinical and educational programs, enhancing both patient care and learner development. Opportunities to incorporate interprofessional education within women's health educational programs across organizations are suggested. This is a resource for medical educators, learners, and practicing clinicians from any field of medicine or any health-care profession.

Co-creating an Art Exhibition on Living Well With Dementia.

This article considers ethical questions raised during development of an online art exhibit for and with people with dementia and their care partners. This article also describes a participant engagement process emphasizing patient autonomy as a means of dismantling stigma, promoting personhood, and stimulating community engagement. This co-creative project suggests the potential for patients' art to spark attitude change and promote reciprocal, regenerative care practices.

"Tis Better to Give Than to Receive?" Health-related Benefits of Delivering Peer Support in Type 2 Diabetes: An Explanatory Sequential Mixed-methods Study.

OBJECTIVES: Existing peer support literature in diabetes has focussed predominantly on the health impact it has on the beneficiaries rather than the benefactors. In this mixed-methods study, we examined the effect of delivering peer support (vs receiving) on glycated hemoglobin (A1C) and diabetes distress (DD) at 3 and 12 months as part of a larger diabetes self-management support randomized controlled trial. Maintenance or improvement of outcomes was expected. We also assessed peer leaders' experiences with the program. METHODS: We utilized a sequential explanatory mixed-methods research design that included 58 adults with diabetes (i.e. peer leaders) who completed a 30-hour training program. Peer leaders (n=52) were matched with participants (adults with type 2 diabetes) and invited to undergo assessments at baseline, 3 months and 12 months. Primary clinical and psychosocial outcomes included A1C and DD, respectively. Secondary outcomes were cardiovascular risk factors and depressive symptoms. After the intervention, 17 peer leaders participated in semistructured interviews about their experience. RESULTS: Peer leaders had a mean age of 57.5±11 years and a long history of diabetes (13.9±11 years); over half were male (53.8%) and married/partnered (55.8%). At baseline, peer leaders were at target for A1C (7.0±0.9% [53±10 mmol/mol]) and reported a low level of DD (1.67±0.52). Of the 43 (82.7%) peer leaders who completed the 12-month study, A1C and DD remained stable over 12 months. Secondary outcomes also remained within the normal range from the start to the end of the intervention. CONCLUSION: Delivering peer support may help maintain glycemic control and DD over the long term.

Profiling "Success": Demographic and Personality Predictors of Effective Peer Leaders in a Diabetes Self-management Intervention.

BACKGROUND: The sociodemographic and personality profiles of effective peer leaders in the context of diabetes self-management interventions are poorly understood. In this study, we explored the demographic and personality characteristics of peer leaders participating in a 12-month, telephone-based type 2 diabetes self-management intervention. METHODS: We used a sequential explanatory mixed-methods research design and recruited 52 peer leaders. Thirty-seven peer leaders had at least 1 participant complete both the baseline and the 12-month assessments. Eligible peer leader candidates were English-speaking adults (≥21 years of age) with diabetes and a self-reported glycated hemoglobin (A1C) level of ≤8% who had access to a phone and transportation and were willing to attend a 30-hour training program. Peer leaders completed a self-report survey assessing sociodemographic characteristics and a Mini-International Personality Item Pool scale measuring the "Big 5" personality traits. After the intervention, 17 peer leaders participated in semistructured interviews on their program experience. We categorized peer leaders as effective if their participants sustained or improved their A1C and diabetes distress (DD) scores from baseline to 12 months, and as ineffective if their participants worsened on any of these parameters. RESULTS: Our cohort scored highest on agreeableness and lowest on neuroticism. Twenty peer leaders were considered effective, most of whom were male, married, employed and educated. They also had significantly lower mean DD levels (p=0.02) and a higher extroversion score (p=0.03) at baseline. CONCLUSIONS: Extroversion emerged as the best personality predictor of peer leader effectiveness. These results, in combination with interview responses, were used to produce a peer leader selection model.

Out of the mouths of Peer Leaders: Perspectives on how to improve a telephone-based peer support intervention in type 2 diabetes.

OBJECTIVE: To explore the experiences of peer leaders with respect to delivering core components of a 12-month, telephone-based peer support intervention in type 2 diabetes within a tertiary-care setting. METHODS: Seventeen peer leaders were recruited and interviewed. Interviews lasted approximately 20 to 45 min, were audio-taped, and transcribed verbatim. The transcripts were analysed by two team members using the qualitative descriptive approach. FINDINGS: Peer leaders reported mutually beneficial and reciprocal relationships with participants. They encountered challenges in maintaining regular contact with participants and in motivating them to make lifestyle changes. To improve the programme, peer leaders suggested having more frequent - but shorter - training sessions and reducing the diabetes education component of the training programme. To enhance the intervention fidelity and retention rate, they recommended matching peer leaders to participants on more meaningful variables (e.g. diabetes-related commonalities, personality, life experiences, etc.) beyond just gender, geographic proximity and availability. They also requested more frequent face-to-face contacts with participants (Modality of Contact), and additional ongoing support from the research team. CONCLUSION: Peer leaders were satisfied with the intervention design. However, future studies may consider more comprehensive peer leader-matching algorithms and increased opportunities for in-person communication modalities. CLINICALTRIALS: gov Identifier: NCT02804620.

Integration of health systems science and women's healthcare.

Health systems science addresses the complex interactions in healthcare delivery. At its core, health systems science describes the intricate details required to provide high-quality care to individual patients by assisting them in navigating the multifaceted and often complicated US healthcare delivery system. With advances in technology, informatics, and communication, the modern physician is required to have a strong working knowledge of health systems science to provide effective, low-cost, high-quality care to patients. Medical educators are poised to introduce health systems science concepts alongside the basic science and clinical science courses already being taught in medical school. Because of the common overlap of women's healthcare subject matter with health systems science topics, such as interprofessional collaboration, ethics, advocacy, and quality improvement, women's health medical educators are at the forefront of incorporating health systems science into the current medical school educational model. Here, the authors have described the concept of health systems science and discussed both why and how it should be integrated into the undergraduate medical education curriculum. Medical educators must develop physicians of the future who can not only provide excellent patient care but also actively participate in the advancement and improvement of the healthcare delivery system.

Research-based Theatre about veterans transitioning home: A mixed-methods evaluation of audience impacts.

RATIONALE: Contact!Unload, a research-based theatre production, portrays veterans experiencing mental health challenges and overcoming them through therapeutic enactment. It was performed eight times by veteran performers in 2017 for audiences in two Canadian cities comprised of civilians and military-connected personnel and their families (n = 525). METHODS: Drawing upon qualitative and quantitative data sources, this paper evaluates the immediate and longer-term impacts of Contact!Unload as a knowledge translation intervention for audience members. RESULTS: Our findings suggest that the performance: 1) improved knowledge of mental health concerns and symptoms that some veterans experience when transitioning to civilian life, 2) increased knowledge of the need for mental health supports and care for veterans, 3) sustained impacts on awareness and knowledge six months after the play and 4) sparked dialogue and actions after the show for some audience members. Moreover, theatre was seen as a powerful medium to engage audience members both cognitively and affectively in the topic. CONCLUSIONS: Research-based theatre has significant potential as a knowledge translation intervention for mental health topics. The work also points to the untapped potential of using RbT to engage audience members in a mental health literacy intervention. Future work is needed to study how to effectively combine research-based theatre with intervention design frameworks and other mental health literacy interventions.