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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.

Human cytomegalovirus (HCMV) is the most common cause of congenital virus infection. Congenital HCMV infection occurs in 0.2-1% of all births, and causes birth defects and developmental abnormalities, including sensorineural hearing loss and developmental delay. Several key studies have established the guinea pig as a tractable model for the study of congenital HCMV infection and have shown that polyclonal antibodies can be protective. In this study, we demonstrate that an anti-guinea pig CMV (GPCMV) glycoprotein H/glycoprotein L neutralizing monoclonal antibody protects against fetal infection and loss in the guinea pig. Furthermore, we have delineated the kinetics of GPCMV congenital infection, from maternal infection (salivary glands, seroconversion, placenta) to fetal infection (fetus and amniotic fluid). Our studies support the hypothesis that a neutralizing monoclonal antibody targeting an envelope GPCMV glycoprotein can protect the fetus from infection and may shed light on the therapeutic intervention of HCMV congenital infection in humans.

In this study, we used bioluminescence imaging (BLI) to track long-term transgene activity following the transfection of brain cells using a nonviral gene therapy technique. Formulations of deoxyribonucleic acid (DNA) combined with 30-mer lysine polymers (substituted with 10 kDa polyethylene glycol) form nanoparticles that transfect brain cells in vivo and produce transgene activity. Here we show that a single intracerebral injection of these DNA nanoparticles (DNPs) into the rat cortex, striatum, or substantia nigra results in long-term and persistent luciferase transgene activity over an 8- to 11-week period as evaluated by in vivo BLI analysis, and single injections of DNPs into the mouse striatum showed stable luciferase transgene activity for 1 year. Compacted DNPs produced in vivo signals 7- to 34-fold higher than DNA alone. In contrast, ex vivo BLI analysis, which is subject to less signal quenching from surrounding tissues, demonstrated a DNP to DNA alone ratio of 76- to 280-fold. Moreover, the ex vivo BLI analysis confirmed that signals originated from the targeted brain structures. In summary, BLI permits serial analysis of luciferase transgene activity at multiple brain locations following gene transfer with DNPs. Ex vivo analysis may permit more accurate determination of relative activities of gene transfer vectors.

We employ both manual and automated content analysis to update an existing dictionary (Boritz, Hayes, and Lim 2013) used to classify types of information technology weakness (ITW). Through analysis of auditors' reports on internal control filed under Section 404 of the Sarbanes-Oxley Act from 2011 through 2020, we provide a robust tool for textual analytics. Additionally, we apply OpenAI's large-language model neural networks for classification as a comparison point. We expand identification of ITW categories such as Design, Security, and Outsourcing and identify Governance as a critical new category in auditor reporting. We further expand the dictionary, capturing categories of IT risk found within current IT control frameworks (e.g., COBIT and AICPA TSCs), which we compare with content of auditor reporting. Evidence suggests stakeholders are subject to significant information asymmetry in assessing and reporting IT risk. We provide direction for future IT governance and improve application of advanced textual analysis. [ABSTRACT FROM AUTHOR]

The purpose of this article is to describe the steps to the development of a successful Physical Education Teacher Education (PETE) workshop. PETE professors share a common goal for improving undergraduate pedagogy instruction. Pedagogy issues and concerns include curriculum decisions, field experience assignments, technology in the classroom, peer teaching ideas, assessment, electronic portfolios and textbook decisions. As a way of sharing ideas and brainstorming, university and college PETE professors in Tennessee meet annually at rotating campus sites. A core group of organizers plan the workshop based on interests and concerns express by attendees. The ultimate goal of the workshops is to better prepare future physical educators who are currently enrolled in university and college undergraduate licensure programs. The Tennessee Association for Health, Physical Education, Recreation and Dance (TAHPERD) sponsors the annual event.

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.

Although immunization against amyloid-[beta] (A[beta]) holds promise as a disease-modifying therapy for Alzheimer disease (AD), it is associated with an undesirable accumulation of amyloid in the cerebrovasculature [i.e., cerebral amyloid angiopathy (CAA)] and a heightened risk of micro-hemorrhages. The central and peripheral mechanisms postulated to modulate amyloid with anti-A[beta] immunotherapy remain largely elusive. Here, we compared the effects of prolonged intracerebroventricular (icv) versus systemic delivery of anti-A[beta] antibodies on the behavioral and pathological changes in an aged Tg2576 mouse model of AD. Prolonged icv infusions of anti-A[beta] antibodies dose-dependently reduced the parenchymal plaque burden, astrogliosis, and dystrophic neurites at doses 10- to 50-fold lower than used with systemic delivery of the same antibody. Both icv and systemic anti-A[beta] antibodies reversed the behavioral impairment in contextual fear conditioning. More importantly, unlike systemically delivered anti-A[beta] antibodies that aggravated vascular pathology, icv-infused antibodies globally reduced CAA and associated micro-hemorrhages. We present data suggesting that the divergent effects of icv-delivered anti-A[beta] antibodies result from gradually engaging the local (i.e., central) mechanisms for amyloid clearance, distinct from the mechanisms engaged by high doses of anti-A[beta] antibodies that circulate in the vasculature following systemic delivery. With robust efficacy in reversing AD-related pathology and an unexpected benefit in reducing CAA and associated micro-hemorrhages, icv-targeted passive immunotherapy offers a promising therapeutic approach for the long-term management of AD. Alzheimer disease | passive immunotherapy | vascular amyloid | microglia | peripheral sink

When doctors speak, Wall Street listens. And Wall Street is actively soliciting physician voices: thousands of doctors are being paid to participate in surveys and act as consultants to hedge [...]

Analyzes a set of undergraduate syllabi for a survey of the history of psychology. Identifies several trends: most instructors rely heavily on textbooks for course organization, three texts accounted for more than 60% of book choices, and newer areas of historical interest did not appear to any substantial degree. (DSK)

You work hard for the money, so it's only logical that you should want to protect it. Although every physician's worst nightmare is being wiped out as a result of [...]

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.

In this audio essay, the author/speaker contemplates the perceived relationship between the voice and the speaker—the person in the voice—and asks the following questions: What does it mean when we judge that the person does not sound like they should? In terms of pedagogy and beyond, what does it mean when we judge a person as not speaking the way we think they should? How does our judgment affect and effect the speaker? [ABSTRACT FROM AUTHOR]

Nine definitions of minimum competency testing (MCT) found in the literature are examined, with major issues for policymakers to consider as they make decisions whether MCT will serve the goals and purposes of their testing programs. Perceived benefits and costs and criticisms based on implementation procedures are discussed. (Author/CM)

The on-going debate of dodgeball as an acceptable physical education activity seems endless. Reeducating physical education teachers toward current best practices has proven a difficult task. Many teachers continue to include dodgeball in their yearly activities and defend it as a curriculum choice by asserting that important basic skills such as dodging, throwing, and catching are part of the game. Other support statements include, "the students enjoy it," "we only use soft balls," or "we modify the rules so children are not eliminated." In this article, the author contends that, although those who defend dodgeball as an appropriate activity can find ways to justify its inclusion in the curriculum according to the current National Standards, these same people might respond differently if asked to write an additional Standard that would eliminate controversy by legitimizing dodgeball and other games like it. The overall purpose of the National Standards is to provide a framework for preparing quality physical education curriculums that result in "physically educated person(s)."

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]