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Abstract Outer membrane proteins (OMPs) in Gram-negative bacteria are essential for a number of cellular functions including nutrient transport and drug efflux. Escherichia coli BamA is an essential component of the OMP β-barrel assembly machinery and a potential novel antibacterial target that has been proposed to undergo large (~15 Å) conformational changes. Here, we explored methods to isolate anti-BamA monoclonal antibodies (mAbs) that might alter the function of this OMP and ultimately lead to bacterial growth inhibition. We first optimized traditional immunization approaches but failed to identify mAbs that altered cell growth after screening >3000 hybridomas. We then developed a “targeted boost-and-sort” strategy that combines bacterial cell immunizations, purified BamA protein boosts, and single hybridoma cell sorting using amphipol-reconstituted BamA antigen. This unique workflow improves the discovery efficiency of FACS + mAbs by >600-fold and enabled the identification of rare anti-BamA mAbs with bacterial growth inhibitory activity in the presence of a truncated lipopolysaccharide layer. These mAbs represent novel tools for dissecting the BamA-mediated mechanism of β-barrel folding and our workflow establishes a new template for the efficient discovery of novel mAbs against other highly dynamic membrane proteins.

Abstract Background BRCA1:c.5017_5019del (p.His1673del) is a founder variant relatively frequent in Northern Italy. Despite previous suggestion of pathogenicity, variant classification in public databases is still conflicting, needing additional evidence. Methods Maximum likelihood penetrance of breast/ovarian and other cancer types was estimated using full pedigree data from 53 informative Italian families. The effect of the variant on BRCA1‐ABRAXAS1 interaction was assessed using a GFP‐fragment reassembly‐based PPI assay. Results were combined with additional data from multiple sources to classify the variant according to ACMG/AMP classification rules specified for BRCA1/2. Results Variant‐carriers displayed increased risk for ovarian cancer (HR = 33.0, 95% CI = 7.0–155.0; cumulative risk at age 70 = 27.6%, 95% CI = 12.6–40.0%) but not for breast cancer (HR = 0.7, 95% CI = 0.2–2.2). An increased risk of uterine cancer (HR = 8.0, 95% CI = 1.03–61.6) emerged, warranting further evaluation. Likelihood‐ratio in favor of pathogenicity was 98898642.82 under assumption of standard BRCA1 breast and ovarian penetrance, and 104240832.84 after excluding breast cancer diagnoses (based on penetrance results). Functional analysis demonstrated that the variant abrogates the BRCA1‐ABRAXAS1 binding, supporting the PS3 code assignment within the ACMG/AMP rule‐based model. Collectively, these findings allowed to classify the variant as pathogenic. Conclusion Pathogenicity of BRCA1:c.5017_5019del(p.His1673del) has been confirmed; however, breast cancer risk in Italian families is not increased, unlike in families from other countries and in carriers of most BRCA1 pathogenic variants. The knowledge of atypical risk profiles for this and other variants will pave the way for personalized management based on specific genotype.

The messenger RNA (mRNA) platform emerged at the forefront of vaccine development during the COVID-19 pandemic, with two mRNA COVID-19 vaccines being among the first authorized globally. These vaccines were developed rapidly. Informed by decades of laboratory research, and proved to be safe and efficacious tools for mitigating the global impact of the COVID-19 pandemic. The mRNA platform holds promise for a broader medical application beyond COVID-19. Herein, we provide an overview of this platform and describe lessons learned from the COVID-19 pandemic to help formulate strategies toward enhancing uptake of future mRNA-based interventions. We identify several strategies as vital for acceptance of an expanding array of mRNA-based vaccines and therapeutics, including education, accurate and transparent information sharing, targeted engagement campaigns, continued investment in vaccine safety surveillance, inclusion of diverse participant pools in clinical trials, and addressing deep-rooted inequalities in access to healthcare. We present findings from the Global Listening Project (GLP) initiative, which draws on quantitative and qualitative approaches to capture perceptions and experiences during the COVID-19 pandemic to help design concrete action plans for improving societal preparedness for future emergencies. The GLP survey (>70,000 respondents in 70 countries) revealed tremendous disparities across countries and sociodemographic groups regarding willingness to accept novel mRNA vaccines and medicines. The comfort in innovations in mRNA medicines was generally low (35%) and was marginally lower among women (33%). The GLP survey and lessons learnt from the COVID-19 pandemic provide actionable insights into designing effective strategies to enhance uptake of future mRNA-based medicines.

Abstract There is a growing recognition that responding to climate change necessitates urban adaptation. We sketch a transdisciplinary research effort, arguing that actionable research on urban adaptation needs to recognize the nature of cities as social networks embedded in physical space. Given the pace, scale and socioeconomic outcomes of urbanization in the Global South, the specificities and history of its cities must be central to the study of how well-known agglomeration effects can facilitate adaptation. The proposed effort calls for the co-creation of knowledge involving scientists and stakeholders, especially those historically excluded from the design and implementation of urban development policies.

Objectives: Racial discrimination is associated with numerous negative health outcomes, including increased risk for depression and anxiety symptoms and substance use. Positive affect toward of one's ethnic or racial group (i.e., ethnic–racial identity affirmation) has been shown to buffer the negative effects of racial discrimination on health outcomes. The extent to which one believes his or her group is valued by others (i.e., positive collective ethnic–racial identity) has also been proposed to be protective. However, to date a limited body of research has examined the moderating effect of collective ethnic–racial identity on health, and among available studies, findings are mixed. Method: African American youth (N = 612; 58.2% female, M grade = 8) completed measures on experiences of discrimination, mood symptoms, substance use, ethnic–racial identity affirmation, and collective ethnic–racial identity (assessed using the Collective Self-Esteem Scale). Results: Controlling for demographic variables and affirmation, a significant main effect was found for collective ethnic–racial identity, such that believing that others viewed your group positively was associated with better health outcomes among African American youth. However, collective ethnic–racial identity was not found to buffer the effects of discrimination on health outcomes. Conclusions: These findings highlight the importance of examining collective ethnic–racial identity and the promotive effect it can have on health outcomes for African Americans. More research is needed to better understand if there are health outcomes in which collective ethnic–racial identity may also mitigate risk as a consequence of racial discrimination. [ABSTRACT FROM AUTHOR]

Background and Objectives: Substance use behaviors have been identified as a risk factor that places juveniles at greater risk for engaging in delinquent behaviors and continual contact with the juvenile justice system. Currently, there is lack of research that explores comorbid factors associated with substance use, such as post-traumatic stress disorder (PTSD) symptoms, that could help identify youth who are at greatest risk. The aim of the present study was to examine if PTSD symptomology moderated the relationship between substance use disorder (SUD) symptoms and externalizing behaviors and commission of a violent crime; hypothesizing that risk would be heightened among youth with elevated SUD and PTSD symptomology compared to those with elevated SUD symptoms but lower PTSD symptoms.Method: The study included 194 predominantly male (78.4%), non-White (74.2%) juvenile justice youth between the ages of 9-18 (M = 15.36). Youth provided responses to assess PTSD symptoms, SUD symptoms, and externalizing behaviors. Commission of a violent crime was based on parole officer report.Results: Findings indicated that SUD symptomology was associated with greater externalizing behaviors at high levels of PTSD symptomology. At low levels of PTSD symptomology, SUD symptoms were inversely associated with externalizing behaviors. An interactive relationship was not observed for commission of violent crimes.Conclusions: Findings suggest that the association between SUD symptoms and externalizing behaviors among juvenile offenders may be best explained by the presence of PTSD symptomology.Scientific Significance: Addressing PTSD rather than SUD symptoms may be a better target for reducing risk for externalizing behaviors among this population of youth (Am J Addict 2019;28:29-35). [ABSTRACT FROM AUTHOR]

Abstract Background Despite significant teaching responsibilities and national accreditation standards, most residents do not receive adequate instruction in teaching methods. Published reports of residents-as-teachers programs vary from brief one-time exposures to curricula delivered over several months. A majority of interventions described are one or two-day workshops with no clear follow-up or reinforcement of skills. A three-year longitudinal teaching skills curriculum was implemented with these goals: 1) deliver an experiential skill-based teaching curriculum allowing all residents to acquire, practice and implement specific skills; 2) provide spaced skills instruction promoting deliberate practice/reflection; and 3) help residents gain confidence in their teaching skills. Methods One hundred percent of internal medicine residents (82/82) participated in the curriculum. Every 10 weeks residents attended a topic-specific experiential skills-based workshop. Each workshop followed the same pedagogy starting with debriefing/reflection on residents’ deliberate practice of the previously taught skill and introduction of a new skill followed by skill practice with feedback. Every year, participants completed: 1) assessment of overall confidence in each skill and 2) retrospective pre-post self-assessment. A post-curriculum survey was completed at the end of 3 years. Results Residents reported improved confidence and self-assessed competence in their teaching skills after the first year of the curriculum which was sustained through the three-year curriculum. The curriculum was well received and valued by residents. Conclusions A formal longitudinal, experiential skills-based teaching skills curriculum is feasible and can be delivered to all residents. For meaningful skill acquisition to occur, recurrent continuous skill-based practice with feedback and reflection is important.

Physician's assistants (PA) are an integral part of hospital teams. To the best of our knowledge, this is the first study to examine the effects of a multidisciplinary hospital-wide communication skills training (CST) workshop on PAs. From November 2017 to November 2019, all participants in the CST workshop were sent a web-based retrospective pre-post survey to measure self-reported attitudes and behaviors related to communicating with patients, CST, and specific skills taught. PA responses were analyzed alone and were compared to non-PAs. Non-PAs were physicians and 1 nurse practitioner. In total, 258 PA and 333 non-PA participants were surveyed for an overall response rate of 25%. Among PAs, in 9 out of 10 domains measured, there was a significant change in self-reported attitudes and behaviors toward communicating with patients, CST, and skills taught ( P

Abstract Pathogenic variants (PVs) in multiple genes are known to increase the risk of early-onset renal cancer (eoRC). However, many eoRC patients lack PVs in RC-specific genes; thus, their genetic risk remains undefined. Here, we determine if PVs in DNA damage response and repair (DDRR) genes are enriched in eoRC patients undergoing cancer risk assessment. Retrospective review of de-identified results from 844 eoRC patients, undergoing testing with a multi-gene panel, for a variety of indications, by Ambry Genetics. PVs in cancer-risk genes were identified in 12.8% of patients—with 3.7% in RC-specific, and 8.55% in DDRR genes. DDRR gene PVs were most commonly identified in CHEK2, BRCA1, BRCA2, and ATM. Among the 2.1% of patients with a BRCA1 or BRCA2 PV,

Abstract Background Providing appropriate levels of autonomy to resident physicians is an important facet of graduate medical education, allowing learners to progress toward the ultimate goal of independent practice. While studies have identified the importance of autonomy to the development of resident physicians, less is known about resident perspectives on their “lived experiences” with autonomy and ways in which clinical educators either promote or undermine it. The current study aims to provide an empirically based practical framework based on resident perspectives through which supervising physicians can attempt to more adequately foster resident physician autonomy. Methods Residents completed open ended surveys followed by facilitated group discussions of their perspectives on autonomy. Qualitative thematic analysis identified key themes in resident definitions of autonomy and how clinical educators either promote or undermine resident autonomy during supervision. Fifty-nine resident physicians representing six different specialties from two institutions participated. Results Learners felt that autonomy was critical to their development as independent physicians. Leading the approach to care, a sense of ownership for patients, and receiving appropriate levels of supervision were identified as key components of autonomy. Attending physicians who promoted this active involvement with patient care were felt to have a strong positive influence on resident autonomy. Autonomy was undermined when decisions were micromanaged and resident input in decision-making process was minimized. Conclusions Fostering autonomy is a critical aspect of medical education. Allowing residents to take the lead in the delivery of patient care while supporting them as important members of the health care team can help to promote resident autonomy in the clinical setting.

Multiple small-molecule inhibitors of the ß-secretase enzyme (BACE1) are under preclinical or clinical investigation for Alzheimer's disease (AD). Prior work has illustrated robust lowering of central amyloid β (A#946;) after acute administration of BACE1 inhibitors. However, very few studies have assessed the overall impact of chronically administered BACE1 inhibitors on brain amyloid burden, neuropathology, and behavioral function in aged preclinical models. We investigated the effects of a potent nonbrain-penetrant BACE1 inhibitor, delivered directly to the brain using intracerebroventricular infusion in an aged transgenic mouse model. Intracerebroventricular infusion of the BACE1 inhibitor (0.3-23.5 μ/d) for 8 weeks, initiated in 17-month-old Tg2576 mice, produced dose-dependent increases in brain inhibitor concentrations (0.2-13 μM). BACE1 inhibition significantly reversed the behavioral deficit in contextual fear conditioning, and reduced brain Aß levels, plaque burden, and associated pathology (e.g., dystrophic neurites), with maximal effects attained with ~ 1 μg/d dose. Strikingly, the BACE1 inhibitor also reversed amyloid pathology below baseline levels (amyloid burden at the start of treatment), without adversely affecting cerebral amyloid angiopathy, microhemorrhages, myelination, or neuromuscular function. Inhibitor-mediated decline in brain amyloid pathology was associated with an increase in microglial ramification. This is the first demonstration of chronically administered BACE1 inhibitor to activate microglia, reverse brain amyloid pathology, and elicit functional improvement in an aged transgenic mouse model. Thus, engagement of novel glial-mediated clearance mechanisms may drive diseasemodifying therapeutic benefit with BACE1 inhibition in AD. [ABSTRACT FROM AUTHOR]

Introduction All physicians must learn comprehensive patient care delivery within the electronic health record (EHR). No studies have considered EHR communication training with an emphasis on clinical efficiency. This curriculum provides a method of teaching clinic efficiency while practicing effective patient communication in any EHR clinical situation. The target audience is resident physicians, fellow physicians, faculty physicians, and physician extenders practicing in a primary care setting where the EHR is present. Methods This curriculum of four separate workshops provides a structured EHR approach while addressing communication strategies for preclinical preparation, rapport building, encounter initiation, agenda setting, and visit closure. The curriculum contains interactive presentations, tools, and an evaluation survey. Presenting efficiency issues with the EHR using the ATTEND mnemonic and agenda setting allows documentation while practicing communication techniques that maximize efficiency. Results Postworkshop surveys revealed that participants felt the workshops were helpful (84%). One measurement of efficiency revealed improvement through decreased number of days to note completion after workshop participation. At the Program Directors Workshop, curriculum value was demonstrated by high attendance, with 94% feeling the workshops provided easily utilizable strategies. Discussion The curriculum utilized only the EPIC EHR but would be generalizable. Future directions could include measurement of effective communication and visit efficiency through direct observation and expanded EHR timing data.

Introduction Faculty and residents strive for appropriate autonomy and entrustment. Initial direct supervision of clinical care gradually shifts to increasing levels of resident independence over time. Faculty members are inconsistent in resident supervision leading to missed opportunities for resident independence. Methods Family medicine faculty workshop participants completed teaching style self-evaluations prior to discussion of clinical examples with excessive or insufficient autonomy. Participants reviewed real resident feedback examples to increase insight into teaching styles. Participants were presented with cases to discuss varying degrees of resident autonomy and entrustment. Learners committed to one specific behavior to calibrate the degree of autonomy they provide. Results Of the faculty, 113 members participated in the workshop with the majority (98%) finding the workshop relevant in helping them to identify strategies for reflecting on their degree of autonomy allowed and to look for appropriate situations for enhancing their resident entrustment. Discussion This interactive workshop provided clear ways for addressing the issue of independence versus control in supervision of patient care. It provided a feedback mechanism for educators who provide too much or too little autonomy for the best resident learning. Additionally, this conversation encouraged participants to engage in self-reflection on the autonomy given to their resident.

BACKGROUND:Maternal depression is common among low-income women enrolled in home visiting programs, yet there is considerable variability in the extent to which it is identified and addressed. This study examines outcomes related to postpartum depression screening, receipt of evidence-based services, and reductions in depressive symptoms among clients of home visiting programs in the Health Resources and Services Administration's Maternal, Infant, and Early Childhood Home Visiting Program Home Visiting Collaborative Improvement and Innovation Network (HV CoIIN), the first U.S. national application of the Institute for Healthcare Improvement's Breakthrough Series (BTS) Model in home visiting programs. METHODS AND FINDINGS:Fourteen home visiting programs in eight states, serving a geographically and racially diverse caseload of pregnant women and new mothers, took part in the HV CoIIN. Women in participating home visiting programs received the intervention strategies implemented by their program during participation in the collaborative. HV CoIIN strategies included specific policies and protocols for depression screening and home visitor response to screening results; home visitor training and supervision; delivery of prevention and treatment interventions; and tracking systems for screening, referral, and follow-up. HV CoIIN's proposed primary outcome was that 85% of women who accessed evidence-based services would experience a 25% reduction in depressive symptoms three months after accessing services. Secondary outcomes included an increased percentage of women who were screened for depression within three months of enrollment or birth, who verbally accepted a referral to evidence-based services, and who received one or more evidence-based service contacts. HV CoIIN resulted in improved symptoms among women who accessed services, from 51.1% to 59.9%. HV CoIIN also improved the percent of women screened for depression, from 83.6% to 96.3%, and those with positive depression screens who accessed evidence-based services, from 41.6% to 65.5%. Home visiting programs in this study were early adopters of quality improvement activities, which may limit the generalizability of these results to other home visiting programs. CONCLUSIONS:Home visiting programs can play an important role in closing gaps in maternal depression identification, referrals, service access, and symptom alleviation. Continuous quality improvement and BTS collaborative methods can be used to improve home visiting services in ways that advance national public health priorities and improve population health outcomes.

Abstract Background Experts suggest observation and feedback is a useful tool for teaching and evaluating medical student communication skills during the clinical years. Failing to do this effectively risks contributing to deterioration of students’ communication skills during the very educational period in which they are most important. While educators have been queried about their thoughts on this issue, little is known about what this process is like for learners and if they feel they get educational value from being observed. This study explored student perspectives regarding their experiences with clinical observation and feedback on communication skills. Methods A total of 125 senior medical students at a U.S. medical school were interviewed about their experiences with observation and feedback. Thematic analysis of interview data identified common themes among student responses. Results The majority of students reported rarely being observed interviewing, and they reported receiving feedback even less frequently. Students valued having communication skills observed and became more comfortable with observation the more it occurred. Student-identified challenges included supervisor time constraints and grading based on observation. Most feedback focused on information gathering and was commonly delayed until well after the observed encounter. Conclusions Eliciting students’ perspectives on the effect of observation and feedback on the development of their communication skills is a unique way to look at this topic, and brings to light many student-identified obstacles and opportunities to maximize the educational value of observation and feedback for teaching communication, including increasing the number of observations, disassociating observation from numerically scored evaluation, training faculty to give meaningful feedback, and timing the observation/feedback earlier in clerkships.

Hospital systems interested in improving patient experience and physician engagement may look to physician communication skills training (CST) as a means of improving both. This study examines a 7.5-hour, multi-specialty, hospital-wide physician CST workshop in a large academic hospital system and its effects on participants’ self-efficacy, attitudes, and behaviors related to communicating with patients. Data was gathered from October 2014 through June 2016 through a web-based questionnaire sent to participants 6-weeks post-workshop which focused on skills taught in the course, attitudes toward communication training, and provider behaviors when communicating with patients. Along with demographic questions, a ten question retrospective pre-post format was used with a 5-point scale for the domains measured. Retrospective pre-post methodology may provide a more accurate assessment of a learners’ self-assessment of skills acquisition. A paired t-test was used to examine changes in participants’ self-efficacy, attitudes, and behaviors toward communicating with patients prior to and 6-weeks post-workshop. Linear regression was used to determine if there were any covariates that explained these changes. A total of 161 responses from 490 participants (21 medical specialties) were collected for a response rate of 32.8%. In 9 out of 10 domains measured, a significant change in self-efficacy, attitudes, and behaviors related to communicating with patients occurred (p