Protein engineering enables a soakable crystal form of human CDK7 primed for high-throughput crystallography and structure-based drug design.

Cyclin dependent kinase 7 (CDK7) is an important therapeutic kinase best known for its dual role in cell cycle regulation and gene transcription. Here, we describe the application of protein engineering to generate constructs leading to high resolution crystal structures of human CDK7 in both active and inactive conformations. The active state of the kinase was crystallized by incorporation of an additional surface residue mutation (W132R) onto the double phosphomimetic mutant background (S164D and T170E) that yielded the inactive kinase structure. A novel back-soaking approach was developed to determine crystal structures of several clinical and pre-clinical inhibitors of this kinase, demonstrating the potential utility of the crystal system for structure-based drug design (SBDD). The crystal structures help to rationalize the mode of inhibition and the ligand selectivity profiles versus key anti-targets. The protein engineering approach described here illustrates a generally applicable strategy for structural enablement of challenging molecular targets.

Reflections on being an insider researcher: a study exploring the experiences of men accessing rural mental health services.

BACKGROUND: An embedded single case-study design was used to explore the experiences of men in rural New Zealand accessing mental health services. It is essential for researchers to acknowledge positionality in case study research and the lead author used reflexive practice to acknowledge his values and beliefs. AIM: To explore and demonstrate the reflexive process of the lead author's position as an inside researcher. DISCUSSION: Three groups were involved in the research: men with mental health challenges, their partners, and mental health clinicians. The article presents the initial research through memos and diarying in the context of current literature. CONCLUSION: Reflexivity is essential for ensuring the research process is complete and biases are identified. Positionality exists on a continuum and it is critical for researchers to be honest with themselves, the topic and the group being investigated, to show respect for the participants and the people they represent, as well as to be committed to revealing the truth. IMPLICATIONS FOR PRACTICE: Insider research has the potential to bridge the gap between academia and practice. It facilitates the transfer of research knowledge directly to practitioners, leading to more evidence-informed decision-making and practice.

What discourses shape and reshape men's experiences of accessing mental health support?

The research employs a single embodied case study design with the aim to examine the discourse of men accessing and receiving mental health support alongside those who are the providers of the support. Three groups of adults were interviewed: men who had experienced mental health problems and had attempted to access mental health support; lay people who supported them, such as partners and non-mental health professionals and professionals, such as mental health nurses, social workers, clinical psychologists and general practitioners. Critical discourse analysis (CDA) is used to identify discourses around three emergent themes: well-being, power and dominance and social capital. Participants seeking mental health support often referred to mental health services as not listening or that what was offered was not useful. A lack of belonging and community disconnectedness was apparent throughout all participant interviews. The study is reported according to the COREQ guidelines.

Tunable Release of Ions from Graphene Oxide Laminates for Sustained Antibacterial Activity in a Biomimetic Environment.

Silver has long been recognized for its potent antimicrobial properties, but achieving a slow and longer-term delivery of silver ions presents significant challenges. Previous efforts to control silver ion dosages have struggled to sustain release for extended periods in biomimetic environments, especially in the presence of complex proteins. This challenge is underscored by the absence of technology for sustaining antimicrobial activity, especially in the context of orthopedic implants where long-term efficacy, extending beyond 7 days, is essential. In this study, the tunable, slow, and longer-term release of silver ions from the two-dimensional (2D) nanocapillaries of graphene oxide (GO) laminates incorporated with silver ions (Ag-GO) for antimicrobial applications are successfully demonstrated. To closely mimic a physiologically relevant serum-based environment, a novel in vitro study model using 100% fetal bovine serum (FBS) is introduced as the test medium for microbiology, biocompatibility, and bioactivity studies. To emulate fluid circulation in a physiological environment, the in vitro studies are challenged with serum exchange protocols on different days. The findings show that the Ag-GO coating can sustainably release silver ions at a minimum dosage of 10 µg cm-2 day-1, providing an effective and sustained antimicrobial barrier for over ten days.

Fragment-Based Discovery of Allosteric Inhibitors of SH2 Domain-Containing Protein Tyrosine Phosphatase-2 (SHP2).

The ubiquitously expressed protein tyrosine phosphatase SHP2 is required for signaling downstream of receptor tyrosine kinases (RTKs) and plays a role in regulating many cellular processes. Genetic knockdown and pharmacological inhibition of SHP2 suppresses RAS/MAPK signaling and inhibit the proliferation of RTK-driven cancer cell lines. Here, we describe the first reported fragment-to-lead campaign against SHP2, where X-ray crystallography and biophysical techniques were used to identify fragments binding to multiple sites on SHP2. Structure-guided optimization, including several computational methods, led to the discovery of two structurally distinct series of SHP2 inhibitors binding to the previously reported allosteric tunnel binding site (Tunnel Site). One of these series was advanced to a low-nanomolar lead that inhibited tumor growth when dosed orally to mice bearing HCC827 xenografts. Furthermore, a third series of SHP2 inhibitors was discovered binding to a previously unreported site, lying at the interface of the C-terminal SH2 and catalytic domains.

Prevalence of Moral Injury, Burnout, Anxiety, and Depression in Healthcare Workers 2 Years in to the COVID-19 Pandemic.

ABSTRACT: COVID-19 has led to marked increases in healthcare worker distress. Studies of these phenomena are often limited to a particular element of distress or a specific subset of healthcare workers. We administered the Moral Injury Symptom Scale for Healthcare Professionals, Copenhagen Burnout Inventory, Patient Health Questionnaire-9, and Generalized Anxiety Disorder-7 via online survey to 17,000 employees of a large academic medical center between December 2021 and February 2022. A total of 1945 participants completed the survey. Across all roles, the prevalence of moral injury, burnout, depression, and anxiety were 40.9%, 35.3%-60.6%, 25.4%, and 24.8%, respectively. Furthermore, 8.1% had been bothered by thoughts that they would be better off dead or of hurting themselves for "several days" or more frequently. Healthcare workers across all roles and practice settings are experiencing unsustainable levels of distress, with 1 in 12 regularly experiencing thoughts of self-harm.

Self-care Perception in Physicians and Patients With Type 2 Diabetes.

Objectives: We sought to assess self-care perception consistency between patients with type 2 diabetes mellitus and their physicians and characterize the psychosocial parameters influencing their respective perceptions. Methods: We conducted a study of 627 patients with type 2 diabetes and their physicians throughout 12 family medicine residency clinics in Texas to analyze their respective perceptions of patient self-care. We utilized linear regression testing. Results: There was a lack of consistency in self-care perception between patients and physicians 31.2% of the time. Patient self-care perception was best predicted by the patients' diabetes distress scores (P<.001) and last HbA1c (P<.001). Conclusion: Measuring a patient's diabetes distress may help physicians better understand a patient's self-care perception. By understanding a patient's self-care perception, the physician may be able to explore these notions of positive or negative self-care perception and even gently correct them when necessary. This may allow physicians to better align themselves with the patient, provide behavioral health interventions, and better target and communicate effective plans. Practical Implications: We suggest periodic assessment of diabetes distress and self-care perception to inform the need for education to improve self-efficacy.

First large-scale study of antimicrobial susceptibility data, and genetic resistance determinants, in Fusobacterium necrophorum highlighting the importance of continuing focused susceptibility trend surveillance.

OBJECTIVES: The objective of the study was to explore antimicrobial resistance gene determinant, and phenotypic antibiotic susceptibility, data for Fusobacterium necrophorum from a collection of UK strains. Antimicrobial resistance genes detected in publicly available assembled whole genome sequences were investigated for comparison. METHODS: Three hundred and eighty five F. necrophorum strains (1982-2019) were revived from cryovials (Prolab). Subsequent to sequencing (Illumina) and quality checking, 374 whole genomes were available for analysis. Genomes were interrogated, using BioNumerics (bioMérieux; v 8.1), for the presence of known antimicrobial resistance genes (ARGs). Agar dilution susceptibility results for 313 F. necrophorum isolates (2016-2021) were also examined. RESULTS: The phenotypic data for the 313 contemporary strains demonstrated potential resistance to penicillin in three isolates, using EUCAST v 11.0 breakpoints, and 73 (23%) strains using v 13.0 analysis. All strains were susceptible to multiple agents using v 11.0 guidance other than clindamycin (n = 2). Employing v 13.0 breakpoints, metronidazole (n = 3) and meropenem (n = 13) resistance were also detected. The tet(O), tet(M), tet(40), aph(3')-III, ant(6)-la and blaOXA-85 ARGs were present in publicly available genomes. tet(M), tet(32), erm(A) and erm(B) were found within the UK strains, with correspondingly raised clindamycin and tetracycline minimum inhibitory concentrations. CONCLUSIONS: Susceptibility to antibiotics recommended for the treatment of F. necrophorum infections should not be assumed. With evidence of potential ARG transmission from oral bacteria, and the detection of a transposon-mediated beta-lactamase resistance determinant in F. necrophorum, surveillance of both phenotypic and genotypic antimicrobial susceptibility trends must continue, and increase.

Validating the TeenHITSS to Assess Child Abuse in Adolescent Populations.

PURPOSE: The purpose of this study is to establish the psychometric properties and diagnostic accuracy of the Teen Hurt-Insult-Threaten-Scream-Sex (TeenHITSS), a 5-item measure used to screen adolescents for family violence in clinical settings. METHODS: Study participants included 251 adolescent participants (n=197 not at-risk subsample; n=56 at-risk subsample), recruited from ambulatory care clinics, a medical center at-risk referral clinic, and area shelters. Participants completed a cross-sectional survey, including the TeenHITSS and Parent-Child Conflict Tactics Scales (CTSPC) questionnaires. We calculated internal reliability, validity, and sensitivity and specificity for the full sample and both subsamples for each screening instrument. RESULTS: Concurrent validity between the TeenHITSS and CTSPC was strong (r=.71, P<.000). We determined an optimal cutpoint based on sensitivity and specificity for correctly identifying abuse victims as a score of one or greater on the TeenHITSS. The TeenHITSS also proved slightly superior to the CTSPC in differentiating between victims and nonvictims of adolescent abuse (AUC=.79 vs .73, respectively). CONCLUSIONS: The TeenHITSS screening tool performed as well as the CTSPC in correctly classifying at-risk and not-at-risk teenagers and offers much greater utility to providers by supplying an actionable cut score. The findings of this study suggest that TeenHITSS is a valid and reliable tool to screen for physical and sexual abuse in children ages 13 years and older in clinical settings and can help health care providers detect adolescent abuse and initiate intervention and prevention of future abuse.

Training Students as Navigators for Patients Experiencing Homelessness.

Background and Objectives: Despite the significant effects of homelessness on health, medical and health professions students rarely receive formal education in caring for individuals experiencing homelessness. We describe the implementation and evaluation of a novel student-run Patient Navigator Program (PNP) and its prerequisite elective that trains students in patient navigation principles specific to homelessness in the local community. METHODS: We analyzed pre- and postsurvey matched responses from students immediately before and after course completion. The survey utilizes the externally-validated instruments Health Professional Attitudes Toward the Homeless Inventory (HPATHI) and the Student-Run Free Clinic Project (SRFCP) survey. We examined differences using paired t tests and Wilcoxon signed-rank tests. Students also completed separate end-of-curriculum evaluation forms assessing satisfaction with the course. RESULTS: After completing the elective, students (n=45) demonstrated improvement in self-assessed attitude towards individuals experiencing homelessness (P=.03), specifically an increase in reported social advocacy (P<.001); and an increase in self-perceived knowledge about (P<.001), efficacy in working with (P=.01), and skills in caring for (P<.001) underserved groups. The elective also received high student satisfaction ratings. CONCLUSIONS: Formal education in patient navigation and caring for individuals experiencing homelessness improves self-assessed preparedness of future health care providers in serving homeless and underserved populations.

Relational and individual stress pathways linking discrimination and ageing cardiometabolic health.

Perceived discrimination is a significant risk factor for worse ageing health outcomes. Yet, the specific individual and relational stress pathways linking discrimination to disease are less understood, especially in the context of cardiometabolic health. We tested family stress and psychophysiological distress (negative affect and high-risk lipid/fat metabolism) as mediators linking perceived discrimination to cardiometabolic morbidity and health appraisal over 20 years for midlife adults. Using data from participants who completed the Biomarker Project (2004-2009) of the Midlife in the U.S. project, and examining data over the study's three waves (1995-1996, 2004-2006, and 2013-2014), we used structural equation modelling to test pathways for participants who reported zero cardiometabolic conditions at baseline (n = 799). Greater Time 1 discrimination was associated with greater Time 2 family strain, which was in turn associated with worse negative affect; worse Time 2 negative affect was associated with worse Time 3 health appraisal; metabolic lipids risk did not serve as an indirect pathway to Time 3 cardiometabolic morbidity (χ2  = 147.74, p < 0.001; RMSEA = 0.056; CFI = 0.902; SRMR = 0.047). The inclusion of family in interventions to mitigate the impact of discrimination may be indicated for promoting cardiometabolic wellness.

Predictors of mood or anxiety problems among transgender individuals seeking hormone therapy.

BACKGROUND: Mental health needs of transgender individuals can be complex with individual, social, and medical factors impacting symptoms. This study examines predictors of mood or anxiety problems among transgender individuals seeking hormone therapy (HT). METHODS: A retrospective chart review was conducted at 2 clinics providing gender-affirming HT. Cross-sectional data from initial patient encounters (N = 311) were used in this study. Bivariate correlations and multiple logistic regression analyses were carried out. RESULTS: Transgender women (TW) were 2.2 times more likely to have mood or anxiety problems while transgender men (TM) were 2.6 times more likely as the number of medical comorbidities increased. For both TW and TM, White race significantly increased the likelihood of mood or anxiety problems. Neither previous nor current HT were associated with mood or anxiety problems for TW and TM. However, receiving multiple gender-affirming procedures decreased the likelihood of mood or anxiety problems for TM. CONCLUSIONS: Gender-affirming care and addressing comorbidities can be important aspects of mental health needs for transgender individuals.

The majority of transgender men and women reported 1 or more chronic health conditions. These health conditions were associated with transgender individuals being more likely to have a mood or anxiety problem. Currently receiving or previously receiving hormonal therapy was not associated with mood or anxiety problems for transgender men or women, but having received 1 or multiple gender-affirming procedures was associated with a decrease in likelihood of having a mood or anxiety problem for transgender men. White race also was associated with increased likelihood of having a mood or anxiety problem for transgender men and women. These results highlight the need for primary care physicians to take a comprehensive approach when dealing with the mental health needs of transgender patients by ensuring that general health care needs are met while receiving gender-affirming care.

A General Framework for Exploring Ethical and Legal Issues in Sports Medicine.

BACKGROUND: Medicolegal dilemmas faced by Sports Medicine providers differ from those of other primary care physicians in type and frequency. Exotic, high-profile challenges are widely covered in the literature and offer guidance on how to navigate challenging situations. However, there is a gap in the literature on how to approach more mundane, but nevertheless common, medicolegal dilemmas. PURPOSE: The purpose of this article is to provide Sports Medicine providers simple tools for determining whether a course of action is legal or ethical and offer Sports Medicine educators a model for teaching these skills. METHODS: We searched U.S. federal and state law using the LexisNexis® database for laws regulating Sports Medicine practice, and PubMed for articles related to physician approaches to Sports Medicine legal and ethical challenges. Searches started with the term sports medicine, and included winnowing terms of ethics, ethical practice, ethical guidelines, law, and legal guidelines. We used current ethical practice guidelines followed by the American Medical Association and described in detail by Beauchamp and Childress. RESULTS: We offer a framework for how to approach legal challenges in Sports Medicine and apply it to four common scenarios that Sports Medicine physicians often face. We further suggest that the profession formally address this literature gap with a standardized curriculum in Sports Medicine law. CONCLUSION: A curriculum using this framework and clinical vignettes provides learners and practitioners with familiarity and confidence when legal and ethical challenges arise in Sports Medicine.

Community-Based HIV and Viral Hepatitis Fellowship Evaluation: Results from a Qualitative Study.

PURPOSE: The UMass Chan Medical School/New England AIDS Education and Training Center Community-Based HIV and Viral Hepatitis Fellowship was launched in 2014 to train physicians and nurse practitioners to become experts in outpatient management of HIV, hepatitis B and C, and latent tuberculosis. The purpose of this study was to identify areas of strength and improvement and understand fellows' perceptions of the program and its impact on their current positions and career trajectories. METHODS: Qualitative study utilizing a semi-structured interview guide with (11) fellowship graduates (8 MDs; 3 NPs). 45 to 60 min interviews were conducted in April and May 2021, recorded and transcribed. Transcripts were analyzed for representative themes and general patterns in the data. RESULTS: Results indicate high satisfaction with the fellowship, which left a positive and indelible impact on their careers and patient care. Fellows highlighted the program's commitment to health equity, its role in transforming them into leaders and advocates for HIV in primary care, and their ability to balance their work and training demands with their personal lives and needs. The fellowship motivated them to become more involved in public health initiatives, serve marginalized communities and reduce their health disparities. They expressed confidence in their ability to independently manage outpatient HIV, viral hepatitis B and C, and latent tuberculosis, and found areas of overlap with their work in primary care. CONCLUSION: As the care of people with HIV becomes more commonplace in primary care clinics, it is imperative that primary care providers receive the necessary training and education to meet this need. Our study of 11 former fellows shows that the Community-Based HIV and Viral Hepatitis Fellowship offers such training, spreads it to other institutions, and can be a model for other programs nationwide.

Implementing Fitness and Nutrition Education in Urban, Underserved, Community-Based Montessori Schools: Challenges and Lessons Learned.

BACKGROUND: Few studies have discussed school-based health programs in Montessori education. Lumin has a network of Montessori elementary schools serving mainly lower income families in Dallas, Texas. Since 2015, our medical school has partnered with Lumin to design and implement fitness and nutrition curricula adherent to Montessori principles. OBJECTIVES: To describe a novel Montessori school-based health program and determine avenues for improvement based on lessons learned. METHODS: Led by medical students with guidance from faculty mentors, the program was developed collaboratively with Lumin leaders based on a critical need in their community and shaped with results from a cross-sectional health needs assessment among Lumin families. Data were collected to measure the impact of the program and a program evaluation was conducted after 5 years of operation to explore curriculum refinement.Results and Lessons Learned: The greatest challenges were recruitment of student volunteers, scheduling and coordination, and garnering community interest for secondary activities (e.g., health fairs). CONCLUSIONS: Despite challenges, this partnership has resulted in a successful program that relies on faculty and student volunteers, incorporates community-based participatory research and service learning concepts, and follows Montessori principles.

Assessing the Factors That Influence Medical Students' Attitudes Toward Working With Medically Underserved Populations.

Introduction: The needs of medically-underserved populations (MUPs) are consistently outpacing the number of physicians caring for them. Medical students' motivations toward working with MUPs consistently decline as they progress through medical school. Given the shortage of doctors caring for MUPs, the objective of our study was to further investigate factors that influence medical students' motivation to work with MUPs while they progress through their education. By identifying these elements, we hope to recommend identified factors within medical education that support the development of more physicians who care for MUPs. Methods: This cross-sectional study is an assessment of medical students at the University of Texas at Southwestern Medical School (UTSW). The study utilized the Medical Student Attitudes Toward the Underserved (MSATU), a validated survey that assesses medical student motivations toward the provision of medical care to MUPs. Surveys were administered at three time points selected to represent key transition points in medical education. Results: There was no significant difference between MSATU scores among the three time points. MSATU scores were higher among students who identified as female, had higher empathy, had higher value placed on teamwork, and had higher community-centeredness. MSATU scores were also higher among students planning to specialize in primary care compared to students planning to specialize in a non-primary care field (P=.239). Conclusion: This study identifies factors associated with high MSATU scores within UTSW medical education, including female identification, higher empathy score, higher emphasis on teamwork, higher community-centeredness, and plans to practice primary care. Additionally, the results support maintenance of MSATU scores across all three time points. Future research should examine individual-level data to determine whether individual students are maintaining their MSATU scores or if individual fluctuations are neutralized by group changes.

The Arabidopsis T-DNA mutant SALK_008491 carries a 14-kb deletion on chromosome 3 that provides rare insights into the plant response to dynamic light stress.

In nature, plants experience rapid changes in light intensity and quality throughout the day. To maximize growth, they have established molecular mechanisms to optimize photosynthetic output while protecting components of the light-dependent reaction and CO2 fixation pathways. Plant phenotyping of mutant collections has become a powerful tool to unveil the genetic loci involved in environmental acclimation. Here, we describe the phenotyping of the transfer-DNA (T-DNA) insertion mutant line SALK_008491, previously known as nhd1-1. Growth in a fluctuating light regime caused a loss in growth rate accompanied by a spike in photosystem (PS) II damage and increased non-photochemical quenching (NPQ). Interestingly, an independent nhd1 null allele did not recapitulate the NPQ phenotype. Through bulk sequencing of a backcrossed segregating F2 pool, we identified an ~14-kb large deletion on chromosome 3 (Chr3) in SALK_008491 affecting five genes upstream of NHD1. Besides NHD1, which encodes for a putative plastid Na+/H+ antiporter, the stromal NAD-dependent D-3-phosphoglycerate dehydrogenase 3 (PGDH3) locus was eradicated. Although some changes in the SALK_008491 mutant's photosynthesis can be assigned to the loss of PGDH3, our follow-up studies employing respective single mutants and complementation with overlapping transformation-competent artificial chromosome (TAC) vectors reveal that the exacerbated fluctuating light sensitivity in SALK_008491 mutants result from the simultaneous loss of PGDH3 and NHD1. Altogether, the data obtained from this large deletion-carrying mutant provide new and unintuitive insights into the molecular mechanisms that function to protect the photosynthetic machinery. Moreover, our study renews calls for caution when setting up reverse genetic studies using T-DNA lines. Although second-site insertions, indels, and SNPs have been reported before, large deletion surrounding the insertion site causes yet another problem. Nevertheless, as shown through this research, such unpredictable genetic events following T-DNA mutagenesis can provide unintuitive insights that allow for understanding complex phenomena such as the plant acclimation to dynamic high light stress.

Lack of awareness of nursing as a career choice for men: A qualitative descriptive study.

AIMS: This research aimed to identify the factors that impact why men do not view nursing as a career choice. DESIGN: Qualitative description was utilized to capture the rich narratives and insight of participants. METHODS: Through convenience sampling, nine New Zealand qualified male nurses within their first three years of practice were recruited. Semi-structured interviews were conducted between November 2019 and January 2020 via zoom from sites across New Zealand. All interviews were recorded and transcribed, with the data analysed using Braun and Clarke's thematic analysis. RESULTS: The findings reflected the experiences of the research participants as they made sense of a predominantly female-dominated work environment both during their undergraduate training and prior to recruitment. The research identified two key themes: The first found that men in nursing experienced isolation due to the societal gendering of nursing influencing the participant's knowledge and understanding of what nursing was, as a career. The second theme found that for participants, nursing was not prominent in their awareness when leaving school and making career choices. CONCLUSION: This research indicates that nursing as a career choice for men is still underpinned by a lack of understanding of the actual role of the nurse and what nurses do, and is more supported by a societal perception that nursing is still a feminized profession. Findings can be used to make recommendations for change in the profession to strengthen diversity in the workforce and redefine the recruitment of men into nursing. IMPACT: This research reviewed the career choices of men in nursing and why they chose nursing as a profession. Understanding the barriers of why men do not consider nursing as a career choice assists with finding strategies in both the clinical and academic environments that can enable greater gender diversity within the nursing profession. NO PATIENT OR PUBLIC CONTRIBUTION: This applies to this research as the focus was on male registered nurses only.