Systemic delivery of mutant huntingtin lowering antisense oligonucleotides to the brain using apolipoprotein A-I nanodisks for Huntington disease.

Efficient delivery of therapeutics to the central nervous system (CNS) remains a major challenge for the treatment of neurological diseases. Huntington disease (HD) is a dominantly inherited neurodegenerative disorder caused by a CAG trinucleotide expansion mutation in the HTT gene which codes for a toxic mutant huntingtin (mHTT) protein. Pharmacological reduction of mHTT in the CNS using antisense oligonucleotides (ASO) ameliorates HD-like phenotypes in rodent models of HD, with such therapies being investigated in clinical trials for HD. In this study, we report the optimization of apolipoprotein A-I nanodisks (apoA-I NDs) as vehicles for delivery of a HTT-targeted ASO (HTT ASO) to the brain and peripheral organs for HD. We demonstrate that apoA-I wild type (WT) and the apoA-I K133C mutant incubated with a synthetic lipid, 1,2-dimyristoyl-sn-glycero-3-phosphocholine, can self-assemble into monodisperse discoidal particles with diameters <20 nm that transmigrate across an in vitro blood-brain barrier model of HD. We demonstrate that apoA-I NDs are well tolerated in vivo, and that apoA-I K133C NDs show enhanced distribution to the CNS and peripheral organs compared to apoA-I WT NDs following systemic administration. ApoA-I K133C conjugated with HTT ASO forms NDs (HTT ASO NDs) that induce significant mHTT lowering in the liver, skeletal muscle and heart as well as in the brain when delivered intravenously in the BACHD mouse model of HD. Furthermore, HTT ASO NDs increase the magnitude of mHTT lowering in the striatum and cortex compared to HTT ASO alone following intracerebroventricular administration. These findings demonstrate the potential utility of apoA-I NDs as biocompatible vehicles for enhancing delivery of mutant HTT lowering ASOs to the CNS and peripheral organs for HD.

Preclinical Development and Characterization of Novel Adeno-Associated Viral Vectors for the Treatment of Lipoprotein Lipase Deficiency.

Lipoprotein lipase deficiency (LPLD) results from mutations within the lipoprotein lipase (LPL) gene that lead to a complete lack of catalytically active LPL protein. Glybera was one of the first adeno-associated virus (AAV) gene replacement therapy to receive European Medicines Agency regulatory approval for the treatment of LPLD. However, Glybera is no longer marketed potentially due to a combination of economical, manufacturing, and vector-related issues. The aim of this study was to develop a more efficacious AAV gene therapy vector for LPLD. Following preclinical biodistribution, efficacy and non-Good Laboratory Practice toxicity studies with novel AAV1 and AAV8-based vectors in mice, we identified AAV8 pVR59. AAV8 pVR59 delivered a codon-optimized, human gain-of-function hLPLS447X transgene driven by a CAG promoter in an AAV8 capsid. AAV8 pVR59 was significantly more efficacious, at 10- to 100-fold lower doses, compared with an AAV1 vector based on Glybera, when delivered intramuscularly or intravenously, respectively, in mice with LPLD. Efficient gene transfer was observed within the injected skeletal muscle and liver following delivery of AAV8 pVR59, with long-term correction of LPLD phenotypes, including normalization of plasma triglycerides and lipid tolerance, for up to 6 months post-treatment. While intramuscular delivery of AAV8 pVR59 was well tolerated, intravenous administration augmented liver pathology. These results highlight the feasibility of developing a superior AAV vector for the treatment of LPLD and provide critical insight for initiating studies in larger animal models. The identification of an AAV gene therapy vector that is more efficacious at lower doses, when paired with recent advances in production and manufacturing technologies, will ultimately translate to increased safety and accessibility for patients.

Prevalence and Correlates of Cardiovascular, Pulmonary, Cancer, and Mental Health Comorbidities Among Adults With HIV Who Smoke.

ABSTRACT: Using data from the D.C. Cohort Longitudinal HIV Study, we examined (a) diagnosed mental health and (b) cardiovascular, pulmonary, or cancer (CPC) comorbidity among adults with HIV who smoked. Among 8,581 adults, 4,273 (50%) smoked; 49% of smokers had mental health, and 13% of smokers had a CPC comorbidity. Among smokers, non-Hispanic Black participants had a lower risk for mental health (prevalence ratio [PR]: 0.69; 95% confidence interval [CI] [0.62-0.76]) but a higher risk for CPC (PR: 1.17; 95% CI [0.84-1.62]) comorbidity. Male participants had a lower risk for mental health (PR: 0.88; 95% CI [0.81-0.94]) and CPC (PR: 0.68; 95% CI [0.57-0.81]) comorbidity. All metrics of socioeconomic status were associated with a mental health comorbidity, but only housing status was associated with a CPC comorbidity. We did not find any association with substance use. Gender, socioeconomic factors, and race/ethnicity should inform clinical care and the development of smoking cessation strategies for this population.

Modelling Aotearoa New Zealand's COVID-19 protection framework and the transition away from the elimination strategy.

For the first 18 months of the COVID-19 pandemic, New Zealand used an elimination strategy to suppress community transmission of SARS-CoV-2 to zero or very low levels. In late 2021, high vaccine coverage enabled the country to transition away from the elimination strategy to a mitigation strategy. However, given negligible levels of immunity from prior infection, this required careful planning and an effective public health response to avoid uncontrolled outbreaks and unmanageable health impacts. Here, we develop an age-structured model for the Delta variant of SARS-CoV-2 including the effects of vaccination, case isolation, contact tracing, border controls and population-wide control measures. We use this model to investigate how epidemic trajectories may respond to different control strategies, and to explore trade-offs between restrictions in the community and restrictions at the border. We find that a low case tolerance strategy, with a quick change to stricter public health measures in response to increasing cases, reduced the health burden by a factor of three relative to a high tolerance strategy, but almost tripled the time spent in national lockdowns. Increasing the number of border arrivals was found to have a negligible effect on health burden once high vaccination rates were achieved and community transmission was widespread.

Calculating incidence of Influenza-like and COVID-like symptoms from Flutracking participatory survey data.

This article describes a new method for estimating weekly incidence (new onset) of symptoms consistent with Influenza and COVID-19, using data from the Flutracking survey. The method mitigates some of the known self-selection and symptom-reporting biases present in existing approaches to this type of participatory longitudinal survey data. The key novel steps in the analysis are: 1) Identifying new onset of symptoms for three different Symptom Groupings: COVID-like illness (CLI1+, CLI2+), and Influenza-like illness (ILI), for responses reported in the Flutracking survey. 2) Adjusting for symptom reporting bias by restricting the analysis to a sub-set of responses from those participants who have consistently responded for a number of weeks prior to the analysis week. 3) Weighting responses by age to adjust for self-selection bias in order to account for the under- and over-representation of different age groups amongst the survey participants. This uses the survey package [22] in R [30]. 4) Constructing 95% point-wise confidence bands for incidence estimates using weighted logistic regression from the survey package [21] in R [28]. In addition to describing these steps, the article demonstrates an application of this method to Flutracking data for the 12 months from 27th April 2020 until 25th April 2021.

Ubiquitination and deubiquitination of 4E-T regulate neural progenitor cell maintenance and neurogenesis by controlling P-body formation.

During embryogenesis, neural stem/progenitor cells (NPCs) proliferate and differentiate to form brain tissues. Here, we show that in the developing murine cerebral cortex, the balance between the NPC maintenance and differentiation is coordinated by ubiquitin signals that control the formation of processing bodies (P-bodies), cytoplasmic membraneless organelles critical for cell state regulation. We find that the deubiquitinase Otud4 and the E3 ligase Trim56 counter-regulate the ubiquitination status of a core P-body protein 4E-T to orchestrate the assembly of P-bodies in NPCs. Aberrant induction of 4E-T ubiquitination promotes P-body assembly in NPCs and causes a delay in their cell cycle progression and differentiation. In contrast, loss of 4E-T ubiquitination abrogates P-bodies and results in premature neurogenesis. Thus, our results reveal a critical role of ubiquitin-dependent regulation of P-body formation in NPC maintenance and neurogenesis during brain development.

Pre-clerkship EPA assessments: a thematic analysis of rater cognition.

BACKGROUND: Entrustable Professional Activities (EPAs) assessments measure learners' competence with an entrustment or supervisory scale. Designed for workplace-based assessment EPA assessments have also been proposed for undergraduate medical education (UME), where assessments frequently occur outside the workplace and may be less intuitive, raising validity concerns. This study explored how assessors make entrustment determinations in UME, with additional specific comparison based on familiarity with prior performance in the context of longitudinal student-assessor relationships. METHODS: A qualitative approach using think-alouds was employed. Assessors assessed two students (familiar and unfamiliar) completing a history and physical examination using a supervisory scale and then thought-aloud after each assessment. We conducted a thematic analysis of assessors' response processes and compared them based on their familiarity with a student. RESULTS: Four themes and fifteen subthemes were identified. The most prevalent theme related to "student performance." The other three themes included "frame of reference," "assessor uncertainty," and "the patient." "Previous student performance" and "affective reactions" were subthemes more likely to inform scoring when faculty were familiar with a student, while unfamiliar faculty were more likely to reference "self" and "lack confidence in their ability to assess." CONCLUSIONS: Student performance appears to be assessors' main consideration for all students, providing some validity evidence for the response process in EPA assessments. Several problematic themes could be addressed with faculty development while others appear to be inherent to entrustment and may be more challenging to mitigate. Differences based on assessor familiarity with student merits further research on how trust develops over time.

Seeing Complexity: Cultural Historical Activity Theory (CHAT) As a Lens for Shared Decision Making.

Shared decision making, a collaborative approach between patient and provider that considers the patient's values and preferences in addition to the scientific evidence, is a complex clinical activity that has not realized its full potential. Gaps in education and training have been cited as barriers to shared decision making, and evidence is inconsistent on effective educational interventions. Because individual agents with their own social and behavioral contexts co-construct a shared decision, the educational approach may need to consider the role of patient agency and sociocultural influences. To address the inherent complexity in shared decision making, the authors identified cultural historical activity theory (CHAT) as a framework for analysis. Although certainly not the only relevant theory, CHAT offers an appropriate lens through which the multivoiced nature of shared decision making can be more clearly appreciated. In this article, the authors demonstrate the application of CHAT as a lens for researchers and educators to examine the complexity of shared decision making. The fictitious case presented in this article describes the use of CHAT with a patient who experiences 2 clinical encounters; during the second, shared decision making takes place. Elements of the case are threaded through the article, demonstrating a sample analysis of the interacting activity systems of the patient and physician and highlighting inherent tensions and contradictions. The authors propose CHAT as a tool for future research around the role of agency in shared decision making and other complex topics and as a framework for design of novel instructional strategies. Although not applicable to all topics and settings, CHAT has significant potential within health professions education.

Academies in Health Professions Education: A Scoping Review.

PURPOSE: Thirty years ago, academies were conceived as a sociocultural approach to revitalize the teaching mission of medical schools and to promote educators' career advancement. The academy movement has grown rapidly and now reaches a broad range of health professions education organizations. The authors conducted a scoping review to map the literature and describe the evidence that guides the formation of new academies and justifies the continuation of existing ones. METHOD: The authors searched MEDLINE (via Ovid), Embase (via Elsevier and Ovid), CINAHL (via EBSCOhost), and Web of Science (via Clarivate Analytics) from inception through March 6, 2020, for publications regarding academy-like organizations. They mapped the relevant literature using logic modeling as an organizing framework and included the mission, resources, activities, output, outcomes, and impact of the included academies. RESULTS: Of the 513 publications identified, 43 met the inclusion criteria, the oldest of which was published in 2000. Most publications were either case reports or perspective/opinion pieces (26, 57.8%), while studies presenting empirical findings were less common (11, 24.4%). Publications showed that academies were diversifying and increasingly were part of a broad range of organizations, including departments, hospitals, health science campuses, and national organizations. The mission, resources, and activities were similar across academies. Evaluation studies were largely limited to process measures, and rigorous studies examining outcomes (i.e., changes in academy participants) and impact on the organization at large were rare. CONCLUSIONS: The increase in the number of academy-related publications parallels the accelerating speed of the academy movement. To sustain this movement, rigorous studies must provide evidence that academies contribute to the revitalization of organizations' teaching mission and bring about an academic culture where educators thrive and where education is a legitimate career path.

A pilot randomized controlled trial of a tailored smoking cessation program for people living with HIV in the Washington, D.C. metropolitan area.

OBJECTIVE: Morbidity and mortality from smoking-related diseases among people living with HIV (PLWH) in the U.S. surpasses that due to HIV itself. Conventional smoking cessation treatments have not demonstrated strong efficacy among PLWH. We conducted a pilot randomized controlled trial (RCT) to evaluate a tailored smoking cessation intervention based on the minority stress model. We compared standard of care counseling (SOC) to a tailored intervention (TI) including one face-to-face counseling session incorporating cognitive behavioral therapy to build resilience, and 30 days of 2-way text messaging. RESULTS: The primary outcome was smoking cessation. Secondary outcomes included cigarettes per day (CPD), exhaled carbon monoxide (CO), and cessation self-efficacy. A total of 25 participants were enrolled (TI:11, SOC:14), and 2 were lost to follow-up. There were no significant differences in quit rates between study groups. However, there was a significantly greater decrease in CPD in the TI versus SOC (13.5 vs. 0.0, p-value:0.036). Additionally, self-efficacy increased in both groups (TI p-value:0.012, SOC p-value:0.049) and CO decreased in both groups (TI p-value: < 0.001, SOC p-value:0.049). This intervention shows promise to support smoking cessation among PLWH. A larger study is needed to fully evaluate the efficacy of this approach. CLINICAL TRIAL: Trial Registration: Retrospectively registered (10/20/2020) NCT04594109.

Intensive Longitudinal Study of the Relationship Between Cigalike E-cigarette Use and Cigarette Smoking Among Adult Cigarette Smokers Without Immediate Plans to Quit Smoking.

INTRODUCTION: This study examined the association between the introduction of an e-cigarette and subsequent change in cigarette smoking among smokers who were not immediately interested in quitting. AIMS AND METHODS: The Moment Study was a 21-day intensive longitudinal study with an online follow-up survey at 30 days. After observing baseline cigarette smoking for 1 week, participants received 10 cigalike e-cigarettes on study days 6 and 13. Participants reported cigarettes per day, e-cigarette puffs per day, and e-cigarette satisfaction using text-message-based surveys. RESULTS: The sample of 96 daily smokers was majority female (53.1%), African American (67.7%), and non-Hispanic (95.8%). When e-cigarettes were provided (day 6), average cigarettes per day dropped by 1.82 cigarettes (p < .0001). The within-person e-cigarette puff effect on daily cigarette smoking was significantly negative (ß = -0.023; p = .005); a participant who consumed 100 more e-cigarette puffs in a day than usual for that person was expected to smoke 2.3 fewer cigarettes that day, but this was only true for non-menthol smokers (p = .006). Smokers older than 45 and those who started smoking at a younger age rated e-cigarettes as less satisfying (ps < .05). Participants with greater than the median reported satisfaction were 6.5 times more likely to use an e-cigarette at follow-up. CONCLUSIONS: Giving e-cigarettes to smokers who did not intend to quit reduced their cigarette smoking on days when they used e-cigarette more frequently, but this relationship did not hold for menthol smokers. Satisfaction with e-cigarette use was predictive of continued use 30 days later. IMPLICATIONS: A greater amount of cigalike e-cigarette use resulted in less smoking among adult daily smokers without immediate plans to quit, but a lack of nicotine delivery and satisfaction for these devices may have limited their utility as a replacement for cigarette smoking, especially among menthol smokers. The global concept of "satisfaction" may be an important driver of e-cigarette use among adult smokers.

Attaining full professor: Women's and men's experiences in medical education.

INTRODUCTION: The underrepresentation of women among senior faculty members in medical education is a longstanding problem. The purpose of this international qualitative investigation was to explore women and men's experiences of attaining full professorship and to investigate why women remain underrepresented among the senior faculty ranks. METHODS: Conducted within a social constructionist orientation, our qualitative study employed narrative analysis. Two female and two male participants working in medical education were recruited from five nations: Australia, Canada, the Netherlands, United Kingdom and United States. All participants held an MD or PhD. During telephone interviews, participants narrated the story of their careers. The five faculty members on the research team were also interviewed. Their narratives were included in analysis, rendering their experiences equal to those of the participants. RESULTS: A total of 24 full professors working in medical education were interviewed (n = 15 females and n = 9 males). While some aspects were present across all narratives (ie personal events, career milestones and facilitating and/or impeding factors), participants' experience of those aspects differed by gender. Men did not narrate fatherhood as a role navigated professionally, but women narrated motherhood as intimately connected to their professional roles. Both men and women narrated career success in terms of hard work and overcoming obstacles; however, male participants described promotion as inevitable, whereas women narrated promotion as a tenuous navigation of social structures towards uncertain outcomes. Female and male participants encountered facilitators and inhibitors throughout their careers but described acting on those experiences differently within the cultural contexts they faced. DISCUSSION: Our data suggest that female and male participants had different experiences of the work involved in achieving full professor status. Understanding these gendered experiences and their impact on career progression is an important advancement for better understanding what leads to the underrepresentation of women among senior faculty members in medical education.

Vector Copy Distribution at a Single-Cell Level Enhances Analytical Characterization of Gene-Modified Cell Therapies.

The ability to deliver transgenes into the human genome using viral vectors is a major enabler of the gene-modified cell therapy field. However, the control of viral transduction is difficult and can lead to product heterogeneity, impacting efficacy and safety, as well as increasing the risk of batch failure during manufacturing. To address this, we generated a novel analytical method to measure vector copy distribution at the single-cell level in a gene-modified, lentiviral-based immunotherapy model. As the limited amount of genomic DNA in a single cell hinders reliable quantification, we implemented a preamplification strategy on selected lentiviral and human gene targets in isolated live single cells, followed by quantification of amplified material by droplet digital PCR. Using a bespoke probability framework based on Bayesian statistics, we show that we can estimate vector copy number (VCN) integers with maximum likelihood scores. Notably, single-cell data are consistent with population analysis and also provide an overall measurement of transduction efficiency by discriminating transduced (VCN ≥ 1) from nontransduced (VCN = 0) cells. The ability to characterize cell-to-cell variability provides a powerful high-resolution approach for product characterization, which could ultimately allow improved control over product quality and safety.

Methylglyoxal couples metabolic and translational control of Notch signalling in mammalian neural stem cells.

Gene regulation and metabolism are two fundamental processes that coordinate the self-renewal and differentiation of neural precursor cells (NPCs) in the developing mammalian brain. However, little is known about how metabolic signals instruct gene expression to control NPC homeostasis. Here, we show that methylglyoxal, a glycolytic intermediate metabolite, modulates Notch signalling to regulate NPC fate decision. We find that increased methylglyoxal suppresses the translation of Notch1 receptor mRNA in mouse and human NPCs, which is mediated by binding of the glycolytic enzyme GAPDH to an AU-rich region within Notch1 3'UTR. Interestingly, methylglyoxal inhibits the enzymatic activity of GAPDH and engages it as an RNA-binding protein to suppress Notch1 translation. Reducing GAPDH levels or restoring Notch signalling rescues methylglyoxal-induced NPC depletion and premature differentiation in the developing mouse cortex. Taken together, our data indicates that methylglyoxal couples the metabolic and translational control of Notch signalling to control NPC homeostasis.

Metadata Correction: Mobile Phone Ownership Is Not a Serious Barrier to Participation in Studies: Descriptive Study.

[This corrects the article DOI: 10.2196/mhealth.8123.].

Mobile Phone Ownership Is Not a Serious Barrier to Participation in Studies: Descriptive Study.

BACKGROUND: Rather than providing participants with study-specific data collection devices, their personal mobile phones are increasingly being used as a means for collecting geolocation and ecological momentary assessment (EMA) data in public health research. OBJECTIVE: The purpose of this study was to (1) describe the sociodemographic characteristics of respondents to an online survey screener assessing eligibility to participate in a mixed methods study collecting geolocation and EMA data via the participants' personal mobile phones, and (2) examine how eligibility criteria requiring mobile phone ownership and an unlimited text messaging plan affected participant inclusion. METHODS: Adult (≥18 years) daily smokers were recruited via public advertisements, free weekly newspapers, printed flyers, and word of mouth. An online survey screener was used as the initial method of determining eligibility for study participation. The survey screened for twenty-eight inclusion criteria grouped into three categories, which included (1) cell phone use, (2) tobacco use, and (3) additional criteria. RESULTS: A total of 1003 individuals completed the online screener. Respondents were predominantly African American (605/1003, 60.3%) (60.4%), male (514/1003, 51.3%), and had a median age of 35 years (IQR 26-50). Nearly 50% (496/1003, 49.5%) were unemployed. Most smoked menthol cigarettes (699/1003, 69.7%), and had a median smoking history of 11 years (IQR 5-21). The majority owned a mobile phone (739/1003, 73.7%), could install apps (86.8%), used their mobile phone daily (89.5%), and had an unlimited text messaging plan (871/1003, 86.8%). Of those who completed the online screener, 302 were eligible to participate in the study; 163 were eligible after rescreening, and 117 were enrolled in the study. Compared to employed individuals, a significantly greater proportion of those who were unemployed were ineligible for the study based on mobile phone inclusion criteria (P<.001); yet, 46.4% (333/717) of the individuals who were unemployed met all mobile phone inclusion criteria. CONCLUSIONS: Inclusion criteria requiring participants to use their personal mobile phones for data collection was not a major barrier to study participation for most respondents who completed the online screener, including those who were unemployed. TRIAL REGISTRATION: ClinicalTrials.gov NCT02261363; https://clinicaltrials.gov/ct2/show/NCT02261363 (Archived by WebCite at http://www.webcitation.org/6wOmDluSt).

Exopolysaccharide production in Caulobacter crescentus: A resource allocation trade-off between protection and proliferation.

Complex and interacting selective pressures can produce bacterial communities with a range of phenotypes. One measure of bacterial success is the ability of cells or populations to proliferate while avoiding lytic phage infection. Resistance against bacteriophage infection can occur in the form of a metabolically expensive exopolysaccharide capsule. Here, we show that in Caulobacter crescentus, presence of an exopolysaccharide capsule provides measurable protection against infection from a lytic paracrystalline S-layer bacteriophage (CR30), but at a metabolic cost that reduces success in a phage-free environment. Carbon flux through GDP-mannose 4,6 dehydratase in different catabolic and anabolic pathways appears to mediate this trade-off. Together, our data support a model in which diversity in bacterial communities may be maintained through variable selection on phenotypes utilizing the same metabolic pathway.

An exploratory analysis of adult daily smokers' experiences using e-cigarettes in smoke-free places.

INTRODUCTION: Evidence indicates that one reason cigarette smokers value e-cigarettes is the ability to use them in places where smoking is not permitted. We sought to: 1) explore adult daily smokers' experiences using e-cigarettes in the context of smoke-free places; and 2) describe smokers' perceptions of bystanders' reactions. METHODS: Twenty adult daily smokers in Washington, DC initiated e-cigarettes for three weeks and completed in semi-structured interviews at the end of each week. All interviews (n=60) were digitally-recorded, transcribed verbatim, imported into NVivo 10.0, and analyzed using thematic analysis methodology. RESULTS: The sample had a mean age of 37.9 years and 18 participants reported having smoked their first cigarette by age 18. Common themes included descriptions of: 1) uncertainty about whether smoke-free policies included e-cigarettes; 2) using e-cigarettes in smoke-free places (e.g. restaurants, workplace, public transit-bus and rail); 3) approaches to e-cigarette use in smoke-free places as part of a complex decision-making process, ranging from testing and establishing the social and spatial boundaries of e-cigarette use, to confining e-cigarette use to inside their home; and 4) favorable, unfavorable, and impartial reactions from bystanders facilitated or impeded e-cigarette use, indicating social approval/social disapproval. CONCLUSIONS: Results suggest a continuum of factors, including smoke-free policies and reactions from bystanders may facilitate or impede e-cigarette use among smokers in environments where a smoke-free imperative is well-established. As e-cigarette use evolves, study findings indicate the importance of the social environment and how it could affect those switching from cigarettes to e-cigarettes.