The BCKDH kinase inhibitor BT2 promotes BCAA disposal and mitochondrial proton leak in both insulin-sensitive and insulin-resistant C2C12 myotubes.

Elevated circulating branched-chain amino acids (BCAAs) have been correlated with the severity of insulin resistance, leading to recent investigations that stimulate BCAA metabolism for the potential benefit of metabolic diseases. BT2 (3,6-dichlorobenzo[b]thiophene-2-carboxylic acid), an inhibitor of branched-chain ketoacid dehydrogenase kinase, promotes BCAA metabolism by enhancing BCKDH complex activity. The purpose of this report was to investigate the effects of BT2 on mitochondrial and glycolytic metabolism, insulin sensitivity, and de novo lipogenesis both with and without insulin resistance. C2C12 myotubes were treated with or without low or moderate levels of BT2 with or without insulin resistance. Western blot and quantitative real-time polymerase chain reaction were used to assess protein and gene expression, respectively. Mitochondrial, nuclei, and lipid content were measured using fluorescent staining and microscopy. Cell metabolism was assessed via oxygen consumption and extracellular acidification rate. Liquid chromatography-mass spectrometry was used to quantify BCAA media content. BT2 treatment consistently promoted mitochondrial uncoupling following 24-h treatment, which occurred largely independent of changes in expressional profiles associated with mitochondrial biogenesis, mitochondrial dynamics, BCAA catabolism, insulin sensitivity, or lipogenesis. Acute metabolic studies revealed a significant and dose-dependent effect of BT2 on mitochondrial proton leak, suggesting BT2 functions as a small-molecule uncoupler. Additionally, BT2 treatment consistently and dose-dependently reduced extracellular BCAA levels without altering expression of BCAA catabolic enzymes or pBCKDHa activation. BT2 appears to act as a small-molecule mitochondrial uncoupler that promotes BCAA utilization, though the interplay between these two observations requires further investigation.

Raine syndrome: Prenatally identified severe craniofacial phenotype with multisuture synostosis and brain abnormalities associated with variants in FAM20C.

Raine syndrome (MIM 259775) is a rare autosomal recessive disorder, first described by Raine et al. in 1989, with an estimated prevalence of <1/1,000,000. This is due to pathogenic variants in FAM20C characterized by osteosclerosis, typical craniofacial features, and brain calcifications. Here, we report a novel variant in FAM20C, describe a uniquely severe craniofacial and CNS phenotype of Raine syndrome, and correlate it with prenatal findings. Fetal phenotyping was based on ultrasound and MRI. Solo exome sequencing was performed from DNA extracted from postmortem skin biopsy. Targeted parental variant testing was subsequently performed. A homozygous missense variant NM_020223.4 (c.1445 G > A (p.Gly482Glu)) was identified in FAM20C associated with Raine syndrome. The infant had the characteristic dysmorphic features seen in Raine syndrome. He had particularly significant CNS manifestations consisting of multisuture craniosynostosis with protrusion of the brain parenchyma through fontanelles and cranial lacunae. Histological sections of the brain showed marked periventricular gliosis with regions of infarction, hemorrhage, and cavitation with global periventricular leukomalacia. Numerous dystrophic calcifications were diffusely present. Here, we demonstrate the identification of a novel variant in FAM20C in an infant with the characteristic features seen in Raine syndrome. The patient expands the characteristic phenotype of Raine syndrome to include a uniquely severe CNS phenotype, first identified on prenatal imaging.

Isolation of a CTX-M-55 (ESBL)-Producing Escherichia coli Strain of the Global ST6448 Clone from a Captive Orangutan in the USA.

The purpose of this study was to determine if orangutans (Pongo spp.) living in captivity at a zoo in Wisconsin were colonized with antimicrobial-resistant bacteria and, if found, to identify underlying genetic mechanisms contributing to their resistant phenotypes. We hypothesize that since antimicrobial-resistant bacteria are so prevalent within humans, the animals could also be carriers of such strains given the daily contact between the animals and the zoo staff that care for them. To test this theory, fecal samples from two orangutans were examined for resistant bacteria by inoculation on HardyCHROM™ ESBL and HardyCHROM™ CRE agars. Isolates were identified using MALDI-TOF mass spectrometry and antimicrobial susceptibility testing was performed using a Microscan autoSCAN-4 System. An isolate was selected for additional characterization, including whole genome sequencing (WGS). Using the Type (Strain) Genome Server (TYGS) the bacterium was identified as Escherichia coli. The sequence type identified was (ST/phylogenetic group/ß-lactamase): ST6448/B1/CTX-M-55.

Gynaecological care of women with chronic pelvic pain: Patient perspectives and care preferences.

OBJECTIVE: To explore the experiences and care preferences of women with chronic pelvic pain, with or without a history of sexual trauma, seeking gynaecological care. DESIGN: Qualitative study. SETTING: Ambulatory endometriosis centre. POPULATION OR SAMPLE: Women aged 18-55 years with chronic pelvic pain. METHODS: Baseline demographics and sexual trauma history were obtained, and participants were assigned to focus groups according to a positive (four groups, 13 participants) or negative (two groups, nine participants) screen for a history of sexual trauma. The focus groups were led by a clinical psychologist and a gynaecological surgeon and consisted of semi-structured interviews. The interviews were audio-recorded and transcribed, and the transcripts were coded in NVivo 12. MAIN OUTCOME MEASURES: Content analysis was used to derive themes according to the participants' own words. RESULTS: Participants with chronic pelvic pain, with or without a history of sexual trauma, experienced delay in diagnosis and repetitive dismissals by clinicians. Participants' experiences of dismissals included: clinicians not listening, insufficient allocation of time to appointments and perceived redundant medical testing (i.e. sexually transmitted infection testing, urine cultures, ultrasounds). Participants identified clinician interactions as pivotal in coping with both pelvic pain and sexual abuse. Participants also provided feedback regarding trauma-informed practices and care delivery specific to patients with chronic pelvic pain. CONCLUSION: Patients with chronic pelvic pain, with or without a history of sexual trauma, report negative experiences when interacting with the healthcare system. They have clear needs and preferences regarding gynaecological care and provide feasible suggestions for improving care delivery.

Critical Care in Placenta Accreta Spectrum Disorders-A Call to Action.

The rising in placenta accreta spectrum (PAS) incidence, highlights the need for critical care allotment for these patients. Due to risk for hemorrhage and possible hemorrhagic shock requiring blood product transfusion, hemodynamic instability and risk of end-organ damage, having an intensive care unit (ICU) with surgical expertise (surgical ICU or equivalent based on institutional resources) is highly recommended. Intensive care units physicians and nurses should be familiarized with intraoperative anesthetic and surgical techniques as well as obstetrics physiologic changes to provide postpartum management of PAS. Validated tools such of bedside point of care ultrasound and viscoelastic tests such as thromboelastogram/rotational thromboelastometry (TEG/ROTEM) are clinically useful in the assessment of hemodynamic status (shock diagnosis, assessment of both fluid responsiveness and tolerance) and transfusion guidance (in patients requiring massive transfusion as opposed to tranditional hemostatic resuscitation) respectively. The future of PAS management lies in the collaborative and multidisciplinary environment. We recommend that women with high suspicion or a confirmed PAS should have a preoperative plan in place and be managed in a tertiary center who is experienced in managing surgically complex cases. KEY POINTS: · The rising in placenta accreta spectrum incidence highlights the need for critical care expertise.. · Emerging tools such as point-of-care ultrasound and thromboelastography/rotational thromboelastometry represent new avenues for real time optimization of hemodynamic and hematological care of patients with PAS.. · Patients with PAS should be referred to a tertiary center having an intensive care unit (ICU) with surgical expertise (or equivalent based on institutional resources)..

Factors associated with county-level mental health during the COVID-19 pandemic.

The objective of this study is to determine county-level factors associated with anxiety, depression, and isolation during the coronavirus disease 2019 (COVID-19) pandemic. This study used daily data from 23,592,355 respondents of a nationwide Facebook-based survey from April 2020 to July 2021, aggregated to the week-county level to yield 212,581 observations. Mental distress prevalences were modeled using weighted linear mixed-effects models with a county random effect. These models revealed that weekly percentages of mental distress were higher in counties with higher unemployment rates, populations, and education levels; higher percentages of females, young adults, individuals with a medical condition, and individuals very worried about their finances and COVID-19; and lower percentages of individuals who were working outside the home, living with children, without health insurance, and Black. Anxiety peaked in April 2020, depression in October 2020, and isolation in December 2020. Therefore, United States counties experienced the mental health effects of the pandemic differently dependent upon their characteristics, and mental distress prevalence varied across time.

Out and in Harm's Way: Sexual Minority Students' Psychological and Physical Health after Institutional Betrayal and Sexual Assault.

In a large random sample of undergraduate university students, we investigated whether sexual minority individuals (i.e., lesbian, gay, and bisexual individuals) experienced different rates of sexual violence victimization (including sexual assault and rape) and subsequent institutional betrayal compared to their heterosexual counterparts, and whether such differences may account for disparities in the psychological and physical health of sexual minorities compared to heterosexuals. In addition to differences in sexual assault victimization rates by gender and sexual orientation, we found differences in rates of institutional betrayal. When non-heterosexual women experienced sexual assault, they experienced a significantly higher rate of institutional betrayal compared to heterosexual women. Overall, greater institutional betrayal was associated with greater negative psychological and physical health outcomes in sexual minorities compared to heterosexual students. These findings reinforce the need for institutional reforms related to the climate and reporting of sexual violence on college campuses, and also identify sexual minority students as a population of particular risk for additional harm by their institutions when they need them most.

Perceptions of Institutional Betrayal Predict Suicidal Self-Directed Violence Among Veterans Exposed to Military Sexual Trauma.

OBJECTIVES: We examined perceptions of institutional betrayal among Veterans exposed to military sexual trauma (MST) and whether perceptions of institutional betrayal are associated with symptoms of posttraumatic stress disorder (PTSD), depression, and suicidal ideation and attempt after MST. METHOD: A total of 49 Veterans with MST completed self-report measures and interviews in a Veterans Health Administration setting. RESULTS: Many participants reported perceptions that a military institution created an environment in which MST seemed common, likely to occur, and did not proactively prevent such experiences. Many participants expressed difficulty reporting MST and indicated that the institutional response to reporting was inadequate. Over two-thirds perceived that the institution had created an environment in which they no longer felt valued or in which continued membership was difficult. Perceptions of institutional betrayal were associated with PTSD symptoms, depressive symptoms, and increased odds of attempting suicide after MST. In contrast, perceptions of institutional betrayal were not associated with post-MST suicidal ideation. Among the subsample of Veterans exposed to military sexual assault, the association between institutional betrayal and PTSD symptoms approached significance. CONCLUSIONS: Perceptions regarding institutional betrayal appear to be highly relevant to MST and its sequelae. These findings underscore the importance of Veterans' perceptions of the military institution's efforts to prevent and respond to MST to individual recovery from sexual trauma. Additional research regarding the association between institutional betrayal and health-related outcomes is needed.

Collusion, torture, and inequality: Understanding the actions of the American Psychological Association as institutional betrayal.

The Hoffman Report (Hoffman et al., 2015) documented devastating information about the American Psychological Association (APA) and the profession of psychology in the United States, prompting a public apology and a formal commitment by APA to correct its mistakes (APA, 2015). In the current article, we utilize betrayal trauma theory (Freyd, 1997), including betrayal blindness (e.g., Freyd, 1996; Tang, 2015) and institutional betrayal (Smith & Freyd, 2014b), to understand and learn from APA's behaviors. We further situate this discussion in the context of inequality, both within APA and in American society generally. We detail how the impact of APA's institutional betrayals extended beyond the organization, its members, and the psychology profession, highlighting the potential for disproportionate harm to minorities, including those who were tortured; Muslims, Middle Easterners, Afghans, and non-Americans who were not tortured; and other minority individuals (Gómez, 2015d). Acknowledging, understanding, and addressing its institutional betrayals offers APA the opportunity to take meaningful corrective and preventive measures. We propose several institutional reparations, including making concrete changes with transparency and conducting self-assessments to inform further needed changes (Freyd & Birrell, 2013). By engaging in institutional courage, APA has the potential to grow into an ethical governing body that fulfills its mission to "advance the creation, communication and application of psychological knowledge to benefit society and improve people's lives" (APA, 2016).

Systemic trauma.

Substantial theoretical, empirical, and clinical work examines trauma as it relates to individual victims and perpetrators. As trauma professionals, it is necessary to acknowledge facets of institutions, cultures, and communities that contribute to trauma and subsequent outcomes. Systemic trauma-contextual features of environments and institutions that give rise to trauma, maintain it, and impact posttraumatic responses-provides a framework for considering the full range of traumatic phenomena. The current issue of the Journal of Trauma & Dissociation is composed of articles that incorporate systemic approaches to trauma. This perspective extends conceptualizations of trauma to consider the influence of environments such as schools and universities, churches and other religious institutions, the military, workplace settings, hospitals, jails, and prisons; agencies and systems such as police, foster care, immigration, federal assistance, disaster management, and the media; conflicts involving war, torture, terrorism, and refugees; dynamics of racism, sexism, discrimination, bullying, and homophobia; and issues pertaining to conceptualizations, measurement, methodology, teaching, and intervention. Although it may be challenging to expand psychological and psychiatric paradigms of trauma, a systemic trauma perspective is necessary on both scientific and ethical grounds. Furthermore, a systemic trauma perspective reflects current approaches in the fields of global health, nursing, social work, and human rights. Empirical investigations and intervention science informed by this paradigm have the potential to advance scientific inquiry, lower the incidence of a broader range of traumatic experiences, and help to alleviate personal and societal suffering.

Exemplar or facilitator: An exploration of the lived experience of nurse mentors supporting the adaptation of internationally educated nurses.

INTRODUCTION: Deficits in nursing workforces have led to major increases in overseas recruitment in many countries. Internationally educated nurses recruited within Ireland must complete an adaptation programme before they can practice nursing, a process contingent on the support from nurse mentors. However, it is becoming progressively difficult to identify nurses willing to act as mentors, threatening viability of overseas nurse recruitment. This research set out to address this problem by exploring the lived experiences of nurse mentors during the adaptation programme. METHODS: An interpretative phenomenological analysis research design was utilised to explore the experiences of nurse mentors. Semi-structured interviews were conducted with a maximum variance sample of 11 nurse mentors. The data were subjected to an Interpretive Phenomenological Analysis (IPA) sequential analytical approach yielding integrative themes. RESULTS: The lived experiences of participants coalesced into three conceptual themes, mentor identity, sustainability, and learner socialisation. Participants who self-identified as 'mentor exemplars' expressed more negative perceptions of the role when compared with nurses who positioned themselves as 'mentor facilitators'. Nurse mentors were challenged by the complexities of intercultural dissonance and insufficient time and training for their mentoring role because of the pressures of clinical service. CONCLUSION: Although mentorship is essential for the integration of internationally educated nurses, the sustainability of the mentorship role is contingent on how it is supported through training and protected within busy clinical environments. Critically, nurse mentors need to learn how to conceptualise their role as one of facilitating the development of others rather than presenting themselves as exemplary role models.

Physiological 4-phenylbutyrate promotes mitochondrial biogenesis and metabolism in C2C12 myotubes.

Type 2 diabetes is characterized by elevated circulating blood metabolites such as glucose, insulin, and branched chain amino acids (BCAA), which often coincide with reduced mitochondrial function. 4-Phenylbutyrate (PBA), an ammonia scavenger, has been shown to activate BCAA metabolism, resolve endoplasmic reticulum (ER) stress, and rescue BCAA-mediated insulin resistance. To determine the effect of PBA on the altered metabolic phenotype featured in type 2 diabetes, the present study investigated the effect of PBA on various metabolic parameters including mitochondrial metabolism and mitochondrial biogenesis. C2C12 myotubes were treated with PBA at 0.5 mM (representing physiologically attainable blood concentrations) or 10 mM (representing physiologically unattainable/proof-of-concept levels) for up to 24 h. Mitochondrial and glycolytic metabolism were assessed via oxygen consumption and extracellular acidification rate, respectively. Mitochondrial content, lipid content, and ER stress were measured by fluorescent staining. Metabolic gene expression was measured by qRT-PCR. Both doses of PBA increased expression of indicators of mitochondrial biogenesis, though only PBA at 0.5 mM increased mitochondrial function and content while 10 mM PBA reduced mitochondrial function and content. PBA at 0.5 mM also rescued reduced mitochondrial function during insulin resistance, though PBA also caused a reduced insulin stimulated pAkt expression during insulin resistance. PBA treatment also increased extracellular BCAA accumulation during insulin resistance despite unchanged pBCKDH expression. Taken together, PBA may increase mitochondrial biogenesis, content, and function in a dose-dependent fashion which may have implications for prevention or treatment of metabolic disease such as insulin resistance.

A Framework for Concussion Management Strategies in Law Enforcement Officers.

Athletic trainers are increasingly utilized in non-traditional settings, such as in law enforcement, where they can contribute to healthcare management, including concussion management of law enforcement officers (LEOs). Despite the prevalence of concussions among LEOs, there is a notable gap in concussion management guidelines for this population. LEOs may lack the education and resources necessary for concussion recognition and proper management. Drawing on advancements in concussion management in athletes and military personnel, here we present a comprehensive framework for concussion management in LEOs encompassing concussion education, a graduated return to duty (RTD) protocol, and considerations for implementation and documentation specific to law enforcement. We also present several barriers and facilitators to implementation. Due to job requirements, it is critical for law enforcement organizations and their medical providers to adopt a concussion management strategy. Without proper concussion management, LEOs may risk subsequent injury and/or suffer from prolonged recovery and adverse long-term outcomes.

Punishment Leads to Greater Sensorimotor Learning But Less Movement Variability Compared to Reward.

When a musician practices a new song, hitting a correct note sounds pleasant while striking an incorrect note sounds unpleasant. Such reward and punishment feedback has been shown to differentially influence the ability to learn a new motor skill. Recent work has suggested that punishment leads to greater movement variability, which causes greater exploration and faster learning. To further test this idea, we collected 102 participants over two experiments. Unlike previous work, in Experiment 1 we found that punishment did not lead to faster learning compared to reward (n = 68), but did lead to a greater extent of learning. Surprisingly, we also found evidence to suggest that punishment led to less movement variability, which was related to the extent of learning. We then designed a second experiment that did not involve adaptation, allowing us to further isolate the influence of punishment feedback on movement variability. In Experiment 2, we again found that punishment led to significantly less movement variability compared to reward (n = 34). Collectively our results suggest that punishment feedback leads to less movement variability. Future work should investigate whether punishment feedback leads to a greater knowledge of movement variability and or increases the sensitivity of updating motor actions.

Dangerous safe havens: institutional betrayal exacerbates sexual trauma.

Research has documented the profound negative impact of betrayal within experiences of interpersonal trauma such as sexual assault (Freyd, 1994, 1996; Freyd, DePrince, & Gleaves, 2007). In the current study of college women (N = 345, 79% Caucasian; mean age = 19.69 years, SD = 2.55), we examined whether institutional failure to prevent sexual assault or respond supportively when it occurs may similarly exacerbate posttraumatic symptomatology-what we call "institutional betrayal." Almost half (47%) of the women reported at least one coercive sexual experience and another 21% reported no coercion, but at least one unwanted sexual experience (total reporting unwanted sexual experiences, N = 233). Institutional betrayal (e.g., creating an environment where these experiences seemed more likely, making it difficult to report these experiences) was reported across different unwanted sexual experiences (47% and 45% of women reporting coercion and no coercion, respectively). Those women who reported institutional betrayal surrounding their unwanted sexual experience reported increased levels of anxiety (R(2) = .10), trauma-specific sexual symptoms (R(2) = .17), dissociation (R(2) = .11), and problematic sexual functioning (R(2) = .12). These results suggest that institutions have the power to cause additional harm to assault survivors.

The Selective LAT1 Inhibitor JPH203 Enhances Mitochondrial Metabolism and Content in Insulin-Sensitive and Insulin-Resistant C2C12 Myotubes.

Population data have shown an association between higher circulating branched-chain amino acids (BCAA) and the severity of insulin resistance in people with diabetes. While several studies have assessed BCAA metabolism as a potential target for regulation, less attention has been paid to the role of L-type amino acid transporter 1 (LAT1), the primary transporter of BCAA in skeletal muscle. The aim of this study was to assess the impact of JPH203 (JPH), a LAT1 inhibitor, on myotube metabolism in both insulin-sensitive and insulin-resistant myotubes. C2C12 myotubes were treated with or without 1 µM or 2 µM JPH for 24 h with or without insulin resistance. Western blot and qRT-PCR were used to assess protein content and gene expression, respectively. Mitochondrial and glycolytic metabolism were measured via Seahorse Assay, and fluorescent staining was used to measure mitochondrial content. BCAA media content was quantified using liquid chromatography-mass spectrometry. JPH at 1 µM (but not 2 µM) increased mitochondrial metabolism and content without inducing changes in mRNA expression of transcripts associated with mitochondrial biogenesis or mitochondrial dynamics. Along with increased mitochondrial function, 1µM treatment also reduced extracellular leucine and valine. JPH at 2 µM reduced pAkt signaling and increased extracellular accumulation of isoleucine without inducing changes in BCAA metabolic genes. Collectively, JPH may increase mitochondrial function independent of the mitochondrial biogenic transcription pathway; however, high doses may reduce insulin signaling.

Characterizing Head Acceleration Events in Law Enforcement Cadets During Subject Control Technique Training.

Law enforcement cadets (LECs) complete weeks of subject control technique training. Similar sport-related combat training has been shown to expose participants to head acceleration events (HAEs) that have potential to result in short- and long-term impairments. The purpose of this study was to describe the number and magnitude of HAEs in LECs throughout their training. 37 LECs (7 females; age = 30.6 ± 8.8 years; BMI = 30.0 ± 6.0) were recruited from a law enforcement organization. Participants wore instrumented mouthguards, which recorded all HAEs exceeding a resultant 5 g threshold for training sessions with the potential for HAEs. Participants completed three defensive tactics (DT) training sessions, a DT skill assessment (DTA), and three boxing sessions. Outcome measures included the number of HAEs, peak linear acceleration (PLA), and peak rotational velocity (PRV). There were 2758 true-positive HAEs recorded across the duration of the study. Boxing sessions accounted for 63.7% of all true-positive HAEs, while DT accounted for 31.4% and DTA accounted for 4.9%. Boxing sessions resulted in a higher number of HAEs per session (F2,28 = 48.588, p < 0.001, ηp2 = 0.776), and higher median PLA (F2,28 = 8.609, p = 0.001, ηp2 = 0.381) and median PRV (F2,28 = 11.297, p < 0.001, ηp2 = 0.447) than DT and DTA. The LECs experience a high number of HAEs, particularly during boxing sessions. Although this training is necessary for job duties, HAE monitoring may lead to modifications in training structure to improve participant safety and enhance recovery.

Automated Analysis of Preceptor Comments: A Pilot Study Using Sentiment Analysis to Identify Potential Student Issues in Experiential Education.

OBJECTIVE: The purpose of this paper is to describe a sentiment analysis program that aids in identifying pharmacy students at risk for progression issues by automatically scoring preceptor comments as positive or negative. METHODS: An R-based program to analyze advanced pharmacy practice experiences and introductory pharmacy practice experiences midpoint evaluation of preceptor comments was piloted in phase 1 by comparing the sentiment analysis algorithm results to human coding. The algorithm was refined in phase 2. In phase 3, the validation phase, the final sentiment analysis algorithm analyzed all midpoint student evaluations (n = 1560). Sentiment scores were generated for each preceptor comment, and correlations were performed between sentiment scores and the quantitative scoring provided on the assessment. RESULTS: In phase 1, agreement between faculty coders and sentiment analysis was 96%, and in phase 2, agreement between the final codes and sentiment analysis was 92.4% once keywords were added to the sentiment dictionary. In phase 3, a total of 3919 comments from 1560 evaluations were analyzed, and overall, the sentiment analysis results aligned with the quantitative data. CONCLUSION: This sentiment analysis algorithm was accurate in capturing positive and negative comments corresponding to pharmacy student performance. Given the accuracy of this preliminary validation for flagging preceptor comments, there are numerous implications when considering the use of sentiment analysis in pharmacy education. Using a sentiment analysis program minimizes the number of qualitative preceptor comments needing review by experiential faculty, as this program can aid in identifying students at risk of progression issues.

Consistency between Household and County Measures of Onsite Schooling during the COVID-19 Pandemic.

The academic, socioemotional, and health impacts of school policies throughout the COVID-19 pandemic have been a source of many questions that require accurate information about the extent of onsite schooling occurring. This article investigates school operational status datasets during the pandemic, comparing (1) self-report data collected nationally on the household level through a Facebook-based survey, (2) county-level school policy data, and (3) a school-level closure status dataset based on phone GPS tracking. The percentage of any onsite instruction within states and counties are compared across datasets from December 2020 to May 2021. Sources were relatively consistent at the state level and for large counties, but key differences were revealed between units of measurement, showing differences between policy and household decisions surrounding children's schooling experiences. The consistency levels across sources support the usage of each of the school policy sources to answer questions about the educational experiences, factors, and impacts related to K-12 education across the nation during the pandemic, but it remains vital to think critically as to which unit of measurement is most relevant to targeted research questions.

Hyperactive mTORC1 in lung mesenchyme induces endothelial cell dysfunction and pulmonary vascular remodeling.

Lymphangioleiomyomatosis (LAM) is a progressive cystic lung disease caused by tuberous sclerosis complex 1/2 (TSC1/2) gene mutations in pulmonary mesenchymal cells, resulting in activation of the mechanistic target of rapamycin complex 1 (mTORC1). A subset of patients with LAM develop pulmonary vascular remodeling and pulmonary hypertension. Little, however, is known regarding how LAM cells communicate with endothelial cells (ECs) to trigger vascular remodeling. In end-stage LAM lung explants, we identified EC dysfunction characterized by increased EC proliferation and migration, defective angiogenesis, and dysmorphic endothelial tube network formation. To model LAM disease, we used an mTORC1 gain-of-function mouse model with a Tsc2 KO (Tsc2KO) specific to lung mesenchyme (Tbx4LME-Cre Tsc2fl/fl), similar to the mesenchyme-specific genetic alterations seen in human disease. As early as 8 weeks of age, ECs from mice exhibited marked transcriptomic changes despite an absence of morphological changes to the distal lung microvasculature. In contrast, 1-year-old Tbx4LME-Cre Tsc2fl/fl mice spontaneously developed pulmonary vascular remodeling with increased medial thickness. Single-cell RNA-Seq of 1-year-old mouse lung cells identified paracrine ligands originating from Tsc2KO mesenchyme, which can signal through receptors in arterial ECs. These ECs had transcriptionally altered genes including those in pathways associated with blood vessel remodeling. The proposed pathophysiologic mesenchymal ligand-EC receptor crosstalk highlights the importance of an altered mesenchymal cell/EC axis in LAM and other hyperactive mTORC1-driven diseases. Since ECs in patients with LAM and in Tbx4LME-Cre Tsc2fl/fl mice did not harbor TSC2 mutations, our study demonstrates that constitutively active mTORC1 lung mesenchymal cells orchestrated dysfunctional EC responses that contributed to pulmonary vascular remodeling.