DNA methylation patterns of FKBP5 regulatory regions in brain and blood of humanized mice and humans.

Humanized mouse models can be used to explore human gene regulatory elements (REs), which frequently lie in non-coding and less conserved genomic regions. Epigenetic modifications of gene REs, also in the context of gene x environment interactions, have not yet been explored in humanized mouse models. We applied high-accuracy measurement of DNA methylation (DNAm) via targeted bisulfite sequencing (HAM-TBS) to investigate DNAm in three tissues/brain regions (blood, prefrontal cortex and hippocampus) of mice carrying the human FK506-binding protein 5 (FKBP5) gene, an important candidate gene associated with stress-related psychiatric disorders. We explored DNAm in three functional intronic glucocorticoid-responsive elements (at introns 2, 5, and 7) of FKBP5 at baseline, in cases of differing genotype (rs1360780 single nucleotide polymorphism), and following application of the synthetic glucocorticoid dexamethasone. We compared DNAm patterns in the humanized mouse (N = 58) to those in human peripheral blood (N = 447 and N = 89) and human postmortem brain prefrontal cortex (N = 86). Overall, DNAm patterns in the humanized mouse model seem to recapitulate DNAm patterns observed in human tissue. At baseline, this was to a higher extent in brain tissue. The animal model also recapitulated effects of dexamethasone on DNAm, especially in peripheral blood and to a lesser extent effects of genotype on DNAm. The humanized mouse model could thus assist in reverse translation of human findings in psychiatry that involve genetic and epigenetic regulation in non-coding elements.

A bipolar disorder-associated missense variant alters adenylyl cyclase 2 activity and promotes mania-like behavior.

The single nucleotide polymorphism rs13166360, causing a substitution of valine (Val) 147 to leucine (Leu) in the adenylyl cyclase 2 (ADCY2), has previously been associated with bipolar disorder (BD). Here we show that the disease-associated ADCY2 missense mutation diminishes the enzyme´s capacity to generate the second messenger 3',5'-cylic adenosine monophosphate (cAMP) by altering its subcellular localization. We established mice specifically carrying the Val to Leu substitution using CRISPR/Cas9-based gene editing. Mice homozygous for the Leu variant display symptoms of a mania-like state accompanied by cognitive impairments. Mutant animals show additional characteristic signs of rodent mania models, i.e., they are hypersensitive to amphetamine, the observed mania-like behaviors are responsive to lithium treatment and the Val to Leu substitution results in a shifted excitatory/inhibitory synaptic balance towards more excitation. Exposure to chronic social defeat stress switches homozygous Leu variant carriers from a mania- to a depressive-like state, a transition which is reminiscent of the alternations characterizing the symptomatology in BD patients. Single-cell RNA-seq (scRNA-seq) revealed widespread Adcy2 mRNA expression in numerous hippocampal cell types. Differentially expressed genes particularly identified from glutamatergic CA1 neurons point towards ADCY2 variant-dependent alterations in multiple biological processes including cAMP-related signaling pathways. These results validate ADCY2 as a BD risk gene, provide insights into underlying disease mechanisms, and potentially open novel avenues for therapeutic intervention strategies.

Improving Identification of Patients Experiencing Homelessness in the Electronic Health Record: A Curated Registry Approach.

BACKGROUND: Identification of persons experiencing homelessness (PEH) within healthcare systems is critical to facilitate patient and population-level interventions to address health inequities. OBJECTIVE: We created an enhanced electronic health record (EHR) registry to improve identification of PEH within a safety net healthcare system. DESIGN: We compared patients identified as experiencing homelessness in 2021, stratified by method of identification (i.e., through registration data sources versus through new EHR registry criteria). MAIN MEASURES: Sociodemographic and clinical characteristics, healthcare utilization, engagement with homeless service providers, and mortality. KEY RESULTS: In total, 10,896 patients met the registry definition of a PEH; 30% more than identified through standard registration processes; 78% were identified through only one data source. Compared with those identified only through registration data, PEH identified through new registry criteria were more likely to be female (42% vs. 25%, p < 0.001), Hispanic/Latinx or Black/African American (30% versus 25% and 25% vs. 18%, p < 0.0001), and Medicaid or Medicare beneficiaries (74% vs. 67% and 16% vs.10%, respectively, p < 0.0001). New data sources also identified a higher proportion of patients: at extremes of age (16% < 18 years and 9% ≥ 65 years vs. 2% and 5%, respectively, p < 0.0001), with increased clinical risk (31% with CRG 6-9 vs. 18%, p < 0.0001), and with a mental health diagnosis (56% vs. 42%, p < 0.0001), and a lower proportion of patients with a substance use diagnosis (39% vs. 54%, p < 0.0001) or criminal justice involvement (8% vs. 15%, p < 0.0001). Newly identified patients were more likely to be engaged in primary care (OR 2.03, 95% CI 1.83-2.26) but less likely to be engaged with homeless service providers (OR 0.70, 95% CI 0.63-0.77). CONCLUSIONS: Commonly utilized methods of identifying PEH within healthcare systems may underestimate the population and introduce reporting biases. Recognizing alternate identification methods may more comprehensively and inclusively identify PEH for intervention.

Analysis of Adsorbed Polyphosphate Changes on Milled Titanium Dioxide, Using Low-Field Relaxation NMR and Photoelectron Spectroscopy.

In this study, changes in the adsorbed amount and surface structure of sodium hexametaphosphate (SHMP) were investigated for aluminum-doped TiO2 pigment undergoing milling. Relaxation NMR was utilized as a potential at-line technique to monitor the effect of milling on surface area and surface chemistry, while XPS was used primarily to consider the dispersant structure. Results showed that considerable amounts of weakly adsorbed SHMP could be removed with washing, and the level of dispersant removal increased with time, highlighting destructive effects of sustained high-energy milling. Nonetheless, there were no significant chemical changes to the dispersant, although increases to the bridging oxygen (BO) peak full width at half-maximum (FWHM) suggested some chemical degradation was occurring with excess milling. Relaxation NMR revealed a number of important features. Results with unmilled material indicated that dispersant adsorption could be tracked with pseudo-isotherms using the relative enhancement rate (Rsp), where the Rsp decreased with dispersant coverage, owing to partial blocking of the quadrupolar surface aluminum. Milled samples were also tracked, with very accurate calibrations of surface area possible from either T1 or T2 relaxation data for systems without dispersant. Behavior was considerably more complicated with SHMP, as there appeared to be an interplay between the dispersant surface coverage and relaxation enhancement from the surface aluminum. Nevertheless, findings highlight that relaxation NMR could be used as a real-time technique to monitor the extent of milling processes, so long as appropriate industrial calibrations can be achieved.

A Mortality Surveillance Collaboration Between a Health System and Public Health Department.

We describe a collaboration between a health system and public health department to create a mortality surveillance system. The collaboration enabled the health system to identify more than six times the number of deaths identified through local system medical records alone. This powerful epidemiological process, combining the nuanced data captured through clinical care in health systems with subsequent data on mortality, drives quality improvement, scientific research, and epidemiology that can be of particular benefit to underserved communities. (Am J Public Health. 2023;113(9):943-946. https://doi.org/10.2105/AJPH.2023.307335).

Validity evidence for a team-leading assessment tool in pediatric emergency resuscitations using video review.

Background: Effective leadership of health care action teams has demonstrated positive influence on team performance and patient care, but there is no consensus on how to assess these skills. We developed a novel team leadership assessment tool for leaders of interprofessional pediatric resuscitation teams and collected validity evidence for this tool using video review. Methods: This was a prospective cohort study from November 2021 to October 2022. A novel team leadership assessment tool was developed using literature review and local expertise and then piloted and refined using medical simulation. Pediatric emergency medicine (PEM) fellows from a single tertiary care pediatric medical center were enrolled, and videos of one medical resuscitation and one trauma resuscitation were collected per fellow each month. Three reviewers underwent reviewer training and then scored the videos using the assessment tool. Raters provided feedback on feasibility and ease of use using a 5-point Likert scale. Inter-rater reliability for the assessment tool using Gwet's agreement coefficient and the association between performance and clinical level of training using generalized linear mixed model were calculated. Results: Twelve PEM fellows enrolled and 146 videos were reviewed. The inter-rater reliability for each domain ranged from 0.45 (p < 0.0001) to 0.59 (p < 0.0001), with the inter-rater reliability of the total score being 0.49 (p < 0.0001). The reviewers' mean ratings of the elements of the tool were as follows: clarity of the domains (4.6/5), the independence of each domain from each other (3.9/5), the ease of use of the 5-point Likert scale (4.5/5), the usefulness of the provided examples for each domain (4.6/5), and the ability to assess each domain without having to rewatch (4.5/5). The tool differentiated between levels of clinical training for two of the six domains (p < 0.02). Conclusions: We developed a novel team leadership assessment tool for pediatric resuscitation team leaders that demonstrated moderate inter-rater reliability. The tool was easy to use and feasible for educators to assess the performance of PEM trainees in complex high-stakes clinical situations.

Neurological outcome following out of hospital cardiac arrest: Evaluation of performance of existing risk prediction models in a UK cohort.

Introduction: Out of hospital cardiac arrest (OHCA) is a common problem. Rates of survival are low and a proportion of survivors are left with an unfavourable neurological outcome. Four models have been developed to predict risk of unfavourable outcome at the time of critical care admission - the Cardiac Arrest Hospital Prognosis (CAHP), MIRACLE2, Out of Hospital Cardiac Arrest (OHCA), and Targeted Temperature Management (TTM) models. This evaluation evaluates the performance of these four models in a United Kingdom population and provides comparison to performance of the Acute Physiology and Chronic Health Evaluation II (APACHE-II) score. Methods: A retrospective evaluation of the performance of the models was conducted over a 43-month period in 414 adult, non-pregnant patients presenting consecutively following non-traumatic OHCA to the five units in our regional critical care network. Scores were generated for each model for where patients had complete data (CAHP = 347, MIRACLE2 = 375, OHCA = 356, TTM = 385). Cerebral Performance Category (CPC) outcome was calculated for each patient at last documented follow up and an unfavourable outcome defined as CPC ⩾ 3. Performance for discrimination of unfavourable outcome was tested by generating receiver operating characteristic (ROC) curves for each model and comparing the area under the curve (AUC). Results: Best performance for discrimination of unfavourable outcome was demonstrated by the high risk group of the CAHP score with an AUC of 0.87 [95% CI 0.83-0.91], specificity of 97.1% [95% CI 93.8-100] and positive predictive value (PPV) of 96.3% [95% CI 92.2-100]. The high risk group of the MIRACLE2 model, which is significantly easier to calculate, had an AUC of 0.81 [95% CI 0.76-0.86], specificity of 92.3% [95% CI 87.2-97.4] and PPV of 95.2% [95% CI 91.9-98.4]. Conclusion: The CAHP, MIRACLE2, OHCA and TTM scores all perform comparably in a UK population to the original development and validation cohorts. All four scores outperform APACHE-II in a population of patients resuscitated from OHCA. CAHP and TTM perform best but are more complex to calculate than MIRACLE2, which displays inferior performance.

Impact of standardization on racial and socioeconomic disparities in non-accidental trauma evaluations in infants in a pediatric emergency department.

BACKGROUND: Studies have illustrated racial and socioeconomic disparities in evaluation of non-accidental trauma (NAT). We aimed to investigate how implementation of a standardized NAT guideline in a pediatric emergency department (PED) impacted racial and socioeconomic disparities in NAT evaluation. RESULTS: 1199 patients (541 pre- and 658 post-guideline) were included for analysis. Pre-guideline, patients with governmental insurance were more likely than those with commercial insurance to have a social work (SW) consult completed (57.4% vs. 34.7%, p < 0.001) and a Child Protective Services (CPS) report filed (33.4% vs. 13.8%, p < 0.001). Post-guideline, these disparities were still present. There were no differences in race, ethnicity, insurance type, or social deprivation index (SDI) in rates of complete NAT evaluations pre- or post-guideline implementation. Overall adherence to all guideline elements increased from 19.0% before guideline implementation to 53.2% after (p < 0.001). CONCLUSION: Implementation of a standardized NAT guideline led to significant increase in complete NAT evaluations. Guideline implementation was not associated with elimination of pre-existing disparities in SW consults or CPS reporting between insurance groups.

Impact of a Remote Virtual Reality Curriculum Pilot on Clinician Conflict Communication Skills.

OBJECTIVES: Conflict management skills are essential for interprofessional team functioning, however existing trainings are time and resource intensive. We hypothesized that a curriculum incorporating virtual reality (VR) simulations would enhance providers' interprofessional conflict communication skills and increase self-efficacy. METHODS: We conducted a randomized controlled pilot study of the Conflict Instruction through Virtual Immersive Cases (CIVIC) curriculum among inpatient clinicians at a pediatric satellite campus. Participants viewed a 30-minute didactic presentation on conflict management and subsequently completed CIVIC (intervention group) or an alternative VR curriculum on vaccine counseling (control group), both of which allowed for verbal interactions with screen-based avatars. Three months following VR training, all clinicians participated in a unique VR simulation focused on conflict management that was recorded and scored using a rubric of observable conflict management behaviors and a Global Entrustment Scale (GES). Differences between groups were evaluated using generalized linear models. Self-efficacy was also assessed immediately pre, post, and 3 months postcurriculum. Differences within and between groups were assessed with paired independent and 2-sample t-tests, respectively. RESULTS: Forty of 51 participants (78%) completed this study. The intervention group (n = 17) demonstrated better performance on the GES (P = .003) and specific evidence-based conflict management behaviors, including summarizing team member's concerns (P = .02) and checking for acceptance of the plan (P = .02), as well as statistical improvements in 5 self-efficacy measures compared with controls. CONCLUSIONS: Participants exposed to CIVIC demonstrated enhanced conflict communication skills and reported increased self-efficacy compared with controls. VR may be an effective method of conflict communication training.

Qualitative Analysis of a Virtual Near-Peer Pediatric Boot Camp Elective.

Objective: To explore fourth-year medical students' experience with a virtual, near-peer facilitated pediatric boot camp through the lens of self-determination theory (SDT). Methods: We developed a virtual pediatric boot camp elective for fourth-year medical students pursuing pediatric residency using Kern's six steps of curriculum development. The two-week virtual elective consisted of facilitated video conferences and small group discussions led by two senior pediatric residents. Semi-structured focus groups were conducted after elective completion. Using SDT as our conceptual framework, we explored participants' experience with the near-peer facilitation of the boot camp. Focus group recordings were transcribed and thematically analyzed using deductive coding for SDT, with inductive coding for themes outside the theory's scope. Saturation was reached after three focus groups. The codebook was iteratively revised through peer debriefing between coders and reviewed by other authors. Credibility was established through member checking. Results: Ninety-two percent of eligible medical students (n = 23/25) participated in the boot camp with attendance ranging from 18-21 students per session. Twelve students (52%) participated in three focus groups. Qualitative analysis identified five major themes. Four themes consistent with SDT emerged: competence, autonomy, relatedness to near-peers, and relatedness to specialty/institution. The learning environment, including the virtual setting, emerged as an additional, non-SDT-related theme. Conclusions: Medical students' experience with our virtual boot camp closely aligned with SDT. Near-peer relatedness emerged as a unique theme which could be further investigated in other aspects of medical student education. Future research could evaluate higher-level learning outcomes from near-peer educational opportunities. Supplementary Information: The online version contains supplementary material available at 10.1007/s40670-021-01466-w.

Vegetarian Diet, Growth, and Nutrition in Early Childhood: A Longitudinal Cohort Study.

OBJECTIVES: The primary objective of this study was to examine the relationships between vegetarian diet and growth, micronutrient stores, and serum lipids among healthy children. Secondary objectives included exploring whether cow's milk consumption or age modified these relationships. METHODS: A longitudinal cohort study of children aged 6 months to 8 years who participated in the TARGet Kids! cohort study. Linear mixed-effect modeling was used to evaluate the relationships between vegetarian diet and BMI z-score (zBMI), height-for-age z-score, serum ferritin, 25-hydroxyvitamin D, and serum lipids. Generalized estimating equation modeling was used to explore weight status categories. Possible effect modification by age and cow's milk consumption was examined. RESULTS: A total of 8907 children, including 248 vegetarian at baseline, participated. Mean age at baseline was 2.2 years (SD 1.5). There was no evidence of an association between vegetarian diet and zBMI, height-for-age z-score, serum ferritin, 25-hydroxyvitamin D, or serum lipids. Children with vegetarian diet had higher odds of underweight (zBMI <-2) (odds ratio 1.87, 95% confidence interval 1.19 to 2.96; P = .007) but no association with overweight or obesity was found. Cow's milk consumption was associated with higher nonhigh-density lipoprotein cholesterol (P = .03), total cholesterol (P = .04), and low-density lipoprotein cholesterol (P = .02) among children with vegetarian diet. However, children with and without vegetarian diet who consumed the recommended 2 cups of cow's milk per day had similar serum lipids. CONCLUSIONS: Evidence of clinically meaningful differences in growth or biochemical measures of nutrition for children with vegetarian diet was not found. However, vegetarian diet was associated with higher odds of underweight.

Characterisation of polyphosphate coated aluminium-doped titania nanoparticles during milling.

This paper investigates the characterisation of alumina-doped titania nanoparticles, milled under high-shear over time, in the presence of sodium hexametaphosphate (SHMP) dispersant. Transmission electron microscopy (TEM) indicated that prolonged milling times led to the formation of 10 nm particle fines which were electrostatically attracted to larger particles, where no change in the crystal structure was observed. Primary particle sizes measured by dynamic light scattering (DLS) and TEM were in agreement and showed no change in primary particle size (∼250 nm) with respect to milling time, however, there was a clear reduction in the magnitude of the slow mode decay associated to aggregates. The TiO2 was found to have an isoelectric point (iep) in the range of pH 3-4.5, where an increase in milling time led to a lower pHiep, indicative of an increase in SHMP coverage, which was further supported by an intensification in phosphorus content measured by X-ray fluorescence (XRF). Phosphorus content and zeta potential analysis before and after centrifugal washing showed that SHMP was partially removed or hydrolysed for the longer milled pigment samples, whereas no change was observed for shorter milled samples. Relaxation NMR was also performed, where enhanced relaxation rates at longer milling times were associated partially to increases in surface area and exposure of Al sites, as well as physicochemical changes to SHMP density and structure. It is thought that extended milling times may lead to hydrolysis or other structural changes of the dispersant from the high energy milling conditions, allowing easier removal after washing for longer milled pigments.

Breast cancer induced nociceptor aberrant growth and collateral sensory axonal branching.

The tumour and neuron interaction has a significant impact upon disease progression and the patients quality of life. In breast cancer patients, it is known that there is an interaction between the tumour microenvironment and the sensory neurons to influence the progression of cancer as well as pain, though these mechanisms still need to be clearly defined. Here it is demonstrated that in a rodent orthotopic model of breast cancer (MDA MB 231) there was an increase in nerve fibre innervation into the tumour microenvironment (protein gene product 9.5), which were calcitonin gene related peptide positive C fibre nociceptors. In contrast, there was a reduction in myelinated nerve fibres (NF200). A sensory neuronal cell line was cultured in response to conditioned media from MDA MB231 and MCF7 as well as vascular endothelial growth factor-A (VEGF-A). All these experimental conditions induced sensory neuronal growth, with increased formation of collateral axonal branches. Furthermore, it was demonstrated that MDA MB231 and VEGF-A induced sensory neuronal sensitisation in response to capsaicin a TRPV1 agonist. MDA MB231 induced neuronal growth was suppressed by VEGFR2 inhibition (ZM323881 and neutralising antibody DC101), in addition both MDA MB231 and VEGF-A induced neurite growth was attenuated by the inhibition of ARP2/3 complex through co-treatment with CK666. This demonstrates that breast cancer can interact with the sensory nervous system to drive neuritogenesis through a VEGF-A/VEGFR2/ARP2/3 mediated pathway.

Self-assembly and applications of ultraconcentrated nanoparticle solutions.

We demonstrate a highly efficient method for concentrating, purifying and separating gold nanoparticles. The method relies on localized density gradients that can be formed at an aqueous | organic phase interface. We show that this method is able to concentrate aqueous gold nanoparticles to the point where confinement leads to variable interparticle separations. Furthermore, the physical properties of the resulting solution are drastically altered when compared to water. For example, densities higher than 4.5 g/cm(3) could be generated without nanoparticle aggregation. As far as we are aware, this is one of the highest reported densities of an aqueous solution at room temperature. Finally, the compositions of the solutions generated are highly dependent on parameters such as particle size and background analyte making this technique highly advantageous for the separation of multimodal NP populations and chemical purification, with 99.5% and >99.9% efficiency, respectively.