Objectives/Goals: The Research Education and Community Health (REACH) coalition proposes to develop the infrastructure for continuous and comprehensive collection of community health data to drive programs, education, and funding priorities across municipal agencies, institutions, and nonprofit organizations in Galveston County. Methods/Study Population: The workgroup through REACH will organize and adopt a comprehensive community health needs assessment that 1) accumulates existing, readily available data for shared use (e.g., Center for Health Care Data at the UT School of Public Health, the Texas Department of State Health Services Center for Health Statistics, and Epic Cosmos, a data aggregation tool, used by UTMB and other health systems to improve patient care); 2) utilizes data collected throughout the community (i.e. non-profits, municipal agencies, and law enforcement); and 3) applies qualitative data from focus groups and/or key informant interviews, so we can hear directly from community members about what their needs are. By doing so, we hope all can benefit from having access to current and relevant data to drive our programs, education, and funding. Results/Anticipated Results: This Community Health Needs Assessment is being coordinated by a diverse workgroup including community organizations, researchers, and policy makers who will benefit from access to current and relevant data. The Galveston Youth Risk Student Survey, completed every three years and most recently in 2024, revealed lingering health and mental health effects of the COVID-19 pandemic on County youth. This highlighted the need for community access to current, accurate, and ongoing data to drive programming, interventions, and education. The REACH Coalition, made up of 23 UTMB Centers and Institutes and 39 community organizations, is spearheading this effort as a part of its mission to facilitate collaborative research, service, and educational efforts. Discussion/Significance of Impact: Collected data will be used to establish and support ongoing, coordinated interventions in response to identified needs. Shared ownership of data and project implementation optimizes resources and reduces gaps and/or redundancy in community programming.

Objectives/Goals: Explore the association of gestational age of LPIs and neurodevelopment delay in children exposed to the neonatal intensive care unit (NICU). Compare between groups if exposure to NICU affects neurodevelopment in children who were born between 34 and 37 weeks of gestation. Methods/Study Population: This is a cross-sectional study design to study the association of gestational age and neurodevelopmental delays in a birth cohort in Puerto Rico of children exposed to the NICU. Their neurodevelopment will be measured with the Bayley III. Statistical analysis will be performed using IBM SPSS Statistics 27.0. Descriptive statistics will be used, and normality distributions among all continuous variables, frequency distribution for categorical variables. We will recruit 30 infants between 18–24 months of age that were born between 34–37 weeks of gestation. Infant will be divided with regard to their exposure to NICU experience. Results/Anticipated Results: We anticipate identifying neurodevelopment delays among children born prematurely between 34 to 37 weeks of gestation. We anticipate that our controlled group will have better outcomes when compared to the controlled expose group. We also expect that gestational age impacts adversely neurodevelopment in children who were born between 34 and 37 weeks of gestation. Discussion/Significance of Impact: Approximately 84% preterm birth are considered LPIs. Prematurity is described as a chronic condition; adverse long-term neurodevelopment consequences. Our study promotes early detection and interventions that can reduce the consequences of the neurodevelopment delays in LPIs.

Objectives/Goals: The NIH Common Fund launched Community Partnerships to Advance Science for Society (ComPASS) to study ways to reduce health disparities by addressing underlying structural factors within communities. ComPASS was designed for community organizations to lead research that addresses community needs. Methods/Study Population: ComPASS awarded five health equity research hubs (Hubs) to provide specialized technical support to ComPASS research projects led by community organizations. Expertise provided by the Hubs to the community-led projects will focus on specific facets of community health, including nutrition access, health care access, and built environment. The Hubs support community-led piloting and testing of structural interventions within community settings by providing subject matter expertise in areas including structural and multilevel intervention study design and methods, implementation science, and community engagement. Results/Anticipated Results: The Hubs will provide expertise and support to the community-led research projects around one or more social determinants of health domains: health care access and quality, education access and quality, economic stability, social and community context, and neighborhood and built environment. The Hubs will help identify strategies for measuring health outcomes and assessing the effects of structural and contextual factors on intervention outcomes. We anticipate the ComPASS program will lead to a better understanding of how structural interventions that leverage multi-sectoral partnerships can advance health equity. Discussion/Significance of Impact: Through community-led research, ComPASS projects are implementing structural interventions to address social determinants and advance health equity. The technical scientific support rooted in health equity provided by the Hubs is essential to the success of these research projects.

Objectives/Goals: Inclusive physical activity (PA) interventions could improve trans and gender diverse (TGD) adolescents’ PA levels, perceived social support from peers and adults, and mental health, but no stakeholder-informed interventions exist. We describe parents’ impressions of TGD adolescent children’s PA experiences and advice for a PA intervention. Methods/Study Population: We conducted individual Zoom interviews with parents of 13- to 20-year-old TGD adolescents (n = 15). All parents were recruited from a gender healthcare clinic. All children self-identified as TGD, an umbrella term including people who have a gender identity different from social expectations for their assigned sex. We asked questions regarding children’s current and historical participation in PA, parents’ perceptions of barriers to PA for their TGD children, and parents’ desired intervention components. After each interview, parent participants were compensated $100 for their time. We analyzed interview transcripts, focusing on insights to incorporate in an intervention. This study was approved by the University’s Institutional Review Board. Results/Anticipated Results: Parents shared rich histories of their children’s PA participation, compounding barriers to PA, numerous benefits of PA, and a range of preferences for program activities and inclusive practices (e.g., safety protocols, training for adult leaders). They also emphasized the need for TGD youth to build social connectedness through PA. Discussion/Significance of Impact: We gathered concrete advice from parents on creating a PA intervention, which we will use to build a social PA program that meets TGD adolescents’ needs. Addressing health disparities and improving PA, social support, and mental health among TGD adolescents will require such stakeholder input to improve upon existing PA and sports opportunities.

Objectives/Goals: Gait performance (GP) and global cognitive functions (GCF) are both critical for maintaining independence and quality of life in senior adults. Recent studies have explored the potential link between GP and GCF, encompassing executive functions. Methods/Study Population: PRISMA guidelines will govern this systematic review. This systematic review synthesizes published research from 2000 to 2024, including peer-reviewed articles, pilot studies, and randomized controlled trials, to examine the relationship between GP (how a person walks and stands) and GCP in older adults. The exclusion criteria will be based on studies focused on physical activities unrelated to balance, meta-analysis, and systematic reviews and those published in languages other than English or Spanish. Results/Anticipated Results: Our preliminary data indicate that gait, or walking speed, is significantly correlated with GCP in older adults, with slower walking associated with poorer global cognition. Specifically, gait speed during dual-task walking shows a strong correlation with working memory (p < .001) and processing speed (p < .05) in individuals aged over 60 years. Gender differences were observed, with women over 80 walking slower than men over 85 years, who walked faster, and women exhibited poorer global cognition than men. Discussion/Significance of Impact: Overall, a gait slowing between 0.68 and 1.1 m per second could predict a marker for the risk of developing dementia, indicating that monitoring gait speed in older adults may provide early warning signs, allowing for timely interventions. Enhancing GP and GCF can improve the quality of life and independence in older adults. Acknowledgments: Research supported by NIH: Award Number HCTRECD R25MD007607 from the NIMHD.

Objectives/Goals: Originally presented at TS24, we would like to discuss follow-up outcomes and preliminary findings“Building Empowerment Through Fitness” – objectives were designed to address the “whole women” in underserved segments of the population in impoverished and/or underprivileged communities with fewer opportunities and promote economic mobility. Methods/Study Population: A qualitative study design to understand unique cultural context and challenges faced by women residing in the Birmingham Housing Authority, regarding physical activity engagement, including sedentarism. The needs assessment approach examined barriers, support systems, and social networks. Semi-structured, pre, mid, and post program focus groups explored challenges and opportunities for health, such as diet and exercise, and are being analyzed by hand with a thematic analysis. The project framework used movement, home repair tools with a life-building/life repair curriculum to influence self-efficacy and program engagement. The study enrollment included 51 women over two cohorts in a 12-week project. Results/Anticipated Results: Preliminary are results showed the women did face heightened health risks due to sedentary lifestyles, educational gaps, and socioenvironmental barriers. We incorporated tool skills, healthy lifestyles, and social/emotional focus group to explore social/emotional perceptions, concerns, and needs. Although our results were on target, there were some unexpected outcomes related to family dynamics, literacy, housing stability, and peer support. Discussion/Significance of Impact: Women are more likely to be sedentary and have poor health. BeFit served as a solution to addressing problems related to economic mobility barriers, poor health, sedentariness, and low self-esteem/self-efficacy. Additionally, exercise/movement programs can be a powerful tool to address disparities and help prioritize health.

Objectives/Goals: Trainees in clinical and translational science (CTS) must learn to effectively communicate their research ideas and findings to a range of audiences. As part of our science communication curriculum, we developed ORAL and WRITTEN science communication rubrics for our trainees to use across their courses and research activities. Methods/Study Population: The Tufts CTS Graduate Program is the training core of the Tufts CTSI and its associated pre- and post-doctoral T32 awards. Approximately 10 trainees with a range of backgrounds (e.g., physicians, medical students, master’s-level researchers, and basic science PhDs) matriculate each year. Faculty members and staff with expertise in science communication and pedagogy formed a committee to develop the rubrics. Because oral and written communication require different skills, we developed separate rubrics for each. We reviewed our current science communication curriculum, reviewed existing communication rubrics, and identified common mistakes students make. Following pilot testing by students and faculty pilot for one semester, we modified the rubrics based on informal feedback. Results/Anticipated Results: Both rubrics include a section to identify the target audience and specific items organized by theme. Oral rubric themes include presentation content, slides, verbal communication, nonverbal communication, and following instructions. Written rubric themes include overall, manuscript/proposal sections, and following instructions. The rubrics serve as feedback tools for faculty and students to evaluate work others produce and as self-evaluation tools. Feedback elements include a 4-point rating for each rubric item, open text feedback for each theme, and an open text holistic assessment. We now use the rubrics in our study design course, which features student presentations of planned research, and in our writing course. We anticipate collecting formal student feedback to further evaluate the rubrics. Discussion/Significance of Impact: Our rubrics can supplement existing science communication training and can be integrated into all CTS coursework and research activities. For future clinical and translational scientists to have the greatest impact, they must learn to effectively communicate findings to multiple audiences, ranging from experts in their field to the general public.

Objectives/Goals: This study aims to evaluate the performance of a third-party artificial intelligence (AI) product in predicting diagnosis-related groups (DRGs) in a community healthcare system. We highlight a use case illustrating how clinical practice leverages AI-predicted information in unexpected yet advantageous ways and assess the AI predictions accuracy and practical application. Methods/Study Population: DRGs are crucial for hospital reimbursement under the prospective payment model. The Mayo Clinic Health System (MCHS), a network of clinics and hospitals serving a substantial rural population in Minnesota and Wisconsin, has recently adopted an AI algorithm developed by Xsolis (an AI-focused healthcare solution provider). This algorithm, a 1D convolutional neural network, predicts DRGs based on clinical documentation. To assess the accuracy of AI-generated DRG predictions for inpatient discharges, we analyzed data from 930 patients hospitalized at MCHS Mankato between March 2 and May 13, 2024. The Xsolis platform provided the top three DRG predictions for the first 48 hours of each inpatient stay. The accuracy of these predictions was then compared against the final billed DRG codes from the hospital’s records. Results/Anticipated Results: In our validation set, Xsolis achieved a top-3 DRG prediction accuracy of 71% at 24 hours and 81% at 48 hours, which is lower than the originally reported accuracy of 81.1% and 83.3%, respectively. Interestingly, discussions with clinical practice leaders revealed that the most valuable information derived from the AI predictions was the expected geometric mean length of stay (GMLOS), which Xsolis was perceived to predict accurately. In the Medicare system, each DRG is associated with an expected GMLOS, a critical factor for efficient hospital flow planning. A subsequent analysis comparing predicted GMLOS with the actual length of stay showed variances of -0.10 days on day 1 and 0.14 days on day 2, indicating a high degree of accuracy and aligning with clinical practice perceptions. Discussion/Significance of Impact: Our research underscores that clinical practice can leverage AI predictions in unexpected yet beneficial ways. While initially focused on DRG prediction, the associated GMLOS emerged as more significant. This suggests that AI algorithm validation should be tailored to specific clinical needs rather than relying solely on generalized benchmarks.

Objectives/Goals: Nearly 42% of adults in the USA have obesity; women are disproportionately affected. Women with obesity that become pregnant are especially prone to developing health condition; thus, aid is needed to achieve appropriate gestational weight gain (GWG). GWG interventions would benefit from examining the effects of social support and stress has on GWG. Methods/Study Population: The proposed study will examine whether expecting mothers’ social support predicts their perceived stress. Also, examine whether receiving aid, in the form of weekly meal delivery, changes perceived stress. Lastly, this study will explore if social support mediates the relationship between meal delivery and perceived stress. The study will use data gathered from a pilot GWG intervention focused on providing pregnant women weekly meal delivery. Participants (N = 14) will complete 2 visits at 16–20- and 35–36-weeks gestation, while receiving weekly meal deliveries. Data from both visits will be used for the purpose of this study. Women’s social support will be measured using the Multidimensional Scale of Perceived Social Support. While stress will be measured via the Perceived Stress Scale. Results/Anticipated Results: Previous behavioral weight loss studies have found that individual who endorse experiencing social support report less perceived stress. Although the relationship has seldomly been examined in pregnant women, similar results are anticipated. Often aid, such as meal delivery, can impact perceived stress and is likely to cause a positive change. To our knowledge, this is the first study to explore whether social support mediates the changes in perceived stress after receiving weekly meal delivery. Therefore, there is no known literature to suggest anticipatory results. Discussion/Significance of Impact: Few studies have examined the association between social support and perceived stress in pregnant women. This study will examine the effects social support has on a pregnant women’s stress. The proposed study will aim to identify if meal delivery changes perceived stress. If so, the study will explore if perceived social support mediates this change.

Objectives/Goals: This study aimed to explore Puerto Rican adolescents’ mental health experiences, focusing on risk and protective factors, and cultural perspectives on mental health. Methods/Study Population: Three focus groups were conducted: two with 20 adolescents aged 12 to 18 years of both genders and one with five adults who work with adolescents. Data were analyzed using interpretative phenomenological analysis. Results/Anticipated Results: The majority of participants in both groups of adolescents identified five key themes: social pressures, barriers to discussing mental health, the impact of social media, coping strategies, and institutional interventions. The adult focus group highlighted adolescent mental health problems, support systems, family and social factors, the church’s role, and recommended interventions. Discussion/Significance of Impact: The findings emphasize that social and familial pressures, mental health stigma, and social media significantly impact adolescent mental health. Coping strategies, such as sports, art, and nature, were also identified. These themes underscore the need for safe, supportive spaces, and targeted approaches to address youth mental health.

Objectives/Goals: Urinary tract infections (UTIs) cause significant morbidity, and many patients require multiple courses of antibiotics increasing the risk of antibiotic resistance. We determined the prevalence of urinary antibiotic heteroresistance (HR), which has been associated with treatment failures in vivo, to three first-line antibiotics for UTIs. Methods/Study Population: Clinical urine Escherichia coli isolates from patients in metropolitan Atlanta, Georgia in August 2023 were collected as part of public health surveillance performed by the CDC-funded, Georgia Emerging Infections Program (EIP). Only the first E. coli isolate collected for each patient was included in this study. Antibiotic susceptibility was determined through medical record review. HR to nitrofurantoin, trimethoprim-sulfamethoxazole, and fosfomycin was determined by population analysis profiling (PAP), where broth dilutions of E. coli were plated on increasing concentrations of the antibiotic. HR was defined as survival of >1 in 106 cfu but fewer than 50% survival at 1X antibiotic breakpoint (bp), resistant as > 50% survival at 1X bp and susceptible as survival of Results/Anticipated Results: Among 355 patients, 21 (5.9%) were resistant or intermediate to nitrofurantoin and 92 (26%) were resistant to trimethoprim-sulfamethoxazole. Antibiotic susceptibility data were missing from 5(1.4%) and 11(3%) of isolates for nitrofurantoin and trimethoprim-sulfamethoxazole, respectively. Susceptibility testing was not routinely performed nor reported for fosfomycin, thus excluded. PAP revealed that of the total 355 isolates, 3(0.84%) were heteroresistant to nitrofurantoin, 17(4.8%) were heteroresistant to trimethoprim-sulfamethoxazole, and 27(7.6%) were heteroresistant to fosfomycin. Of the isolates found to be susceptible by standard testing, 1(0.3%) and 9(3.6%) were heteroresistant to nitrofurantoin and trimethoprim-sulfamethoxazole by PAP, respectively. Discussion/Significance of Impact: Despite low rates of HR to nitrofurantoin and trimethoprim-sulfamethoxazole (0.84%, 4.8%), HR to fosfomycin was more frequent (7.6%). Given that susceptibility is not generally performed for fosfomycin, this could have implications for including fosfomycin as a first-line treatment for E. coli UTIs.

Objectives/Goals: Our team has developed a 24-well donor-derived skeletal muscle microphysiological system (MPS) to study signaling pathways associated with a variety of muscle diseases. 3D muscle will be utilized to evaluate pharmacologic interventions for these muscle conditions to improve both muscle mass and function. Methods/Study Population: In this study, muscle MPS were formed from healthy young female and male subjects. 3D muscle underwent a 21-day differentiation with an electrical stimulation (e-stim) regimen twice daily beginning on Day 14. Functional assessments in permeabilized fibers of both sexes included isometric and isotonic calcium-induced contractions, allowing for the characterization of force-pCa (-log[Ca2+]), force-velocity and force-power relationships. Samples from Day 17 and Day 21 will be assessed for pro-growth protein signaling via western blotting and a subset of samples will be analyzed by histology and microscopy for fiber type and size. Finally, culture media pre- and post-terminal e-stim on Day 21 will be collected for extracellular vesicle (EV) isolation and EVs will be assessed by standard proteomics analysis. Results/Anticipated Results: Permeabilized fibers from both sexes reproduced the well-established sigmoidal force-pCa and the curvilinear force-velocity and force-power relationships reported in native striated muscle. Maximum specific force and force-pCa relationship were not different between sexes. Isotonic contractile measurements revealed that these male and female fibers also exhibit similar force-velocity and force-power relationships. We anticipate that 3D muscles from day 17 compared to day 21 will exhibit higher levels of pro-growth protein signaling due to e-stim application and no differences in fiber type or size between sexes. Additionally, we expect that EV quantity will depend upon 3D muscle maturity and presence of e-stim. Discussion/Significance of Impact: This study demonstrates the similarities of functional characteristics and exercise (or e-stim) adaptation between native human skeletal muscle and 3D bioengineered skeletal muscle. Ultimately, this data further validates the muscle MPS system to study muscle diseases and to enhance the translation of therapeutics to clinical settings.

Objectives/Goals: We investigate how the gut microbiome protects against arsenic toxicity, showing antibiotic perturbation increases toxicity and causes interindividual susceptibility to a sepsis-like disease state in mice. Here, we aim to understand how baseline microbiomes from various mouse vendors impact these outcomes and characterize the observed disease. Methods/Study Population: We developed a novel mouse model where mice are exposed to an antibiotic (cefoperazone) for 2 days, followed by co-exposure to the antibiotic and 100 ppm arsenate. So far, we have evaluated C57BL/6N mice from MSU’s in-house colony, Taconic Biosciences (TAC), and Jackson Labs (JAX), along with C57BL/6J mice from JAX. To determine if the baseline microbiome drives inter-vivarium differences, we established in-house breeding colonies of TAC- and JAX-origin mice at MSU. This allowed us to assess whether, when housed under identical conditions, these mice still show differences in mortality based on their original microbiomes. To characterize the arsenic-induced sepsis-like disease, we performed blood biochemistry assays to quantify the white blood cell populations, and sepsis biomarkers used in clinical settings. Results/Anticipated Results: We observed differences in survival rates between genetically identical mice from MSU (45%), TAC (30%), and JAX (2.5%) in our model. From this, we characterized the baseline composition of the gut microbiomes of these mice and found they were significantly different from each other. We are still awaiting results from our in-house TAC and JAX experiments but expect them to have similar gut microbiome compositions to those directly purchased from TAC and JAX and respond similarly. In our blood biochemistry analysis, we found sick mice presented with low WBC counts and notable biomarkers indicative of liver, heart, and kidney distress. We also anticipate that 16S sequencing results of cecal contents will further support findings by providing evidence of a bacterial infection in the ceca of sick mice. Discussion/Significance of Impact: Collectively, our work demonstrates that antibiotic perturbation of the gut microbiome induces an inter-individual and inter-vivarium susceptibility to an arsenic-induced sepsis-like disease state. This work highlights the importance of considering antibiotic use in the risk assessment of arsenic to better protect the health of those exposed.

Objectives/Goals: We aim to explore the associations of race/ethnicity and socioeconomic status (SES): 1) with grip strength, walking speed, and comorbidity index cross-sectionally and 2) with the change in comorbidity index and mortality risk over four years of follow-up in cancer survivors. Both aims will examine the potential mediating role of cytomegalovirus (CMV) infection. Methods/Study Population: This study includes 1,602 cancer survivors (mean age = 72 years, 10% Black, 54% female) from the Health and Retirement Study (HRS), a nationally representative U.S. sample followed for health outcomes until 2020. HRS measured CMV immunoglobulin G (IgG) antibody levels (from blood samples), grip strength, and walking speed in 2016. We will apply linear regression to examine the associations of race/ethnicity and SES with grip strength, walking speed, and comorbidity index cross-sectionally and with the change in comorbidity index over four years of follow-up. We will apply Cox proportional hazard regression to examine the associations of race/ethnicity and SES with mortality over four years of follow-up. In all models, we will investigate the potential mediating role of CMV infection in these associations. Results/Anticipated Results: We expect that CMV infection mediates the associations of race/ethnicity and SES with age-related health outcomes, including muscle weakness (measured by grip strength), decreased functional performance (measured by walking speed), comorbidity index, and mortality in elderly cancer survivors. Discussion/Significance of Impact: If our hypothesis is confirmed, the findings may inform physicians to closely monitor CMV infection among cancer survivors from socially disadvantaged groups and apply treatment if needed. Several oral medications for CMV exist, and CMV vaccines are currently undergoing testing in clinical trials. This will make the treatment for CMV more accessible.

Objectives/Goals: Diffusion of innovations (DoI) posits that new health-related ideas spread through communities over time and across stages of adoption. We will apply DoI to understand a paradigm shift toward seeing participants as partners in clinical research, specifically through delivery of plain language summaries of results. Methods/Study Population: The return of results in lay language (plain language) to clinical trial participants represents a paradigm shift in the EU and now the USA. We will conduct a systematic review of the implementation of “lay summaries” or “plain language summaries” in different jurisdictions to understand current regulatory influence. We will then review PLS samples and published studies to determine the rate of adoption by industry and non-pharmaceutical company sponsors. Using the DoI framework, groups will be placed in an adopter category. Finally, we will employ an implementation science approach to understand the diffusion process and the translation to participants, laying the groundwork for a culture of change in medical product development. Results/Anticipated Results: Our search on PubMed using key terms “Diffusion of Innovation” and “Plain Language Summary” did not produce any relevant results in the context of clinical trials in the USA, which illustrates a gap in the literature and application of this theory in this context. In the future analyses, we will examine factors influencing the adoption stage and outcomes, such as regulatory action, what best practices have been defined/implemented (if any), culture shifts in the context of clinical research, health communications, and inclusion of patient voices in clinical research. Our analysis will include a network analysis to evaluate characteristics that influence adoption of PLS in clinical research. We hope to identify who is at the forefront of innovation and why. Discussion/Significance of Impact: Novel application of DoI theory will help lay the groundwork for a culture of change in patient-focused drug development, specifically for the dissemination of results to patients. In future studies, we plan to develop a tailored framework for the inclusion of PLS as part of a paradigm shift in the patient-focused drug development process.

Objectives/Goals: The goal of this study is to resolve the complexity of the adipose precursor cells and identify potential therapeutic targets/mechanism to treat obesity, diabetes, cancer cachexia, and related metabolic conditions in both white and brown adipose tissues. Methods/Study Population: White and brown adipose precursor cells were isolated from neonatal P0 mice and expanded in culture for single-cell RNA sequencing analysis. Unsupervised machine learning was used to unbiasedly cluster and categorize sequenced cells. Differentially expressed genes were used to identify populations via pathway analysis. Populations were then validated with published datasets via integration, reference mapping, and module scoring to ensure our dataset is reflective of known literature. Then, white and brown datasets were combined and unbiasedly clustered. Finally, signaling inferences using CellChat was used to identify significant signals being sent to and received from each cluster based on ligand–receptor pairs. Results/Anticipated Results: ScRNAseq revealed 7 subclusters in both white and brown adipose tissues. Differential expression and trajectory inferences revealed that white and brown precursors develop into two distinct fates: committed adipogenic precursors (CAPs), where these cells will be mature lipid-laden adipocytes; or fibro-adipogenic precursors-like (FAPLs), where these cells preferably stay in a fibroblast-like, antiadipogenic phenotype. Integrating white and brown cells with subsequent reclustering reveals that white and brown FAPLs are highly similar to one another by being clustered together. Cell signaling inferences and pathway analysis reveal that white and brown FAPLs may participate in the regulation of adipogenesis and angiogenesis of the adipose tissue. Discussion/Significance of Impact: Our results demonstrated that brown and white precursor cells share a common regulatory subpopulation with similar gene expression profiles, highlighting a more interconnected regulatory landscape in adipose tissue than previously understood. These findings reveal novel mechanisms of systemic metabolism and provide new therapeutic targets.

Objectives/Goals: We investigated the risk of trauma in the form of fractures and traumatic brain injuries (TBIs) among Medicare beneficiaries with incident Parkinson’s disease (PD) age ≥67 compared to population-based controls. Secondarily, we examined the risk of death following a fracture in PD cases compared to controls. Methods/Study Population: We identified incident PD cases (N = 94,317) within a population-based sample of 2017 Medicare beneficiaries. Controls (N = 471,585) were matched 5:1 on month and year. We obtained claims data from 2017 to 2019 to follow cases and controls to identify new fractures treated in a hospital. Our primary outcome was any fracture. We also considered fracture type and TBI. We compared frailty level between cases and controls. We used logistic regression models to estimate odds ratios (ORs) and 95% confidence intervals (CIs) for the association between trauma and PD after adjusting for the following covariates: selected medical comorbidities, age, sex, race/ethnicity, smoking, and use of care. We used Cox regression to estimate hazard ratios (HRs) and 95% CI for trauma in cases compared to controls using the same covariates. Results/Anticipated Results: Compared to controls, PD patients who developed a fracture were more likely to have a history of falls (OR = 2.20, 95% CI 2.08–2.34) and difficulties in walking (OR = 2.66, 95% CI 2.50–2.82). Compared to controls with a fracture, PD patients with a fracture were more likely to be moderately frail (OR = 1.43, 95% CI 1.25–1.64). PD cases had a higher risk of all fracture types, including hip (OR = 1.93, 95% CI 1.85, 2.01), spine (OR = 1.90, 95% CI 1.79, 2.02), upper extremity (OR = 1.69, 95% CI 1.58–1.80), and other traumas such as a TBI (OR = 2.14, 95% CI 1.88–2.43). PD patients had greater mortality following a fracture (HR = 1.18, 95% CI 1.13–1.24) than controls. Discussion/Significance of Impact: The burden of trauma in the first two years immediately after PD diagnosis is high and warrants the initiation of early fall and fracture prevention strategies, in addition to aggressive treatment of PD symptoms by all providers caring for patients with PD.

Objectives/Goals: The overall goal of this project is to determine bacterial transcriptional signatures from clinical sputum and assess their potential to monitor treatment response and predict the outcome of drug therapy in patients with tuberculosis (TB). Methods/Study Population: We are developing a novel transcript capture sequencing (TC-Seq) approach to sequence the mRNA of Mycobacterium tuberculosis (Mtb) and analyze transcriptomes from clinical samples containing minimal amounts of bacterial RNA. This protocol generates single-stranded biotinylated probes from Mtb DNA. Probes are hybridized to and allow enrichment of Mtb-specific mRNA within next-generation RNA sequencing libraries. We will apply TC-Seq to sputum samples collected throughout an 18-month Phase II clinical trial investigating response to TB treatment to compare the transcriptome of Mtb between patients whose treatment results in cure or relapse. Results/Anticipated Results: We have refined a technique to generate biotinylated probes starting from DNA of lab grown Mtb. This protocol achieves robust and unbiased sampling of the Mtb transcriptome from mixed samples containing both human and Mtb RNA. Preliminary sequencing of clinical sputum collected pretreatment has generated 1–4 million Mtb-specific reads, a sequencing depth that allows examination of the entire bacterial transcriptome. We will measure differential gene expression before and during treatment as well as between cure and relapse cases. These results will allow us to characterize bacterial response to treatment and identify bacterial markers that correlate with relapse. Discussion/Significance of Impact: Understanding Mtb activity during treatment will offer new ways to assess the efficacy of different treatment regimens. Crucially, identifying clear bacterial markers that demarcate a cure or relapse outcome will have a significant impact on determining patient eligibility for shorter drug therapy.

Objectives/Goals: Intra-amniotic inflammation (IAI) is one of the leading causes of maternal/fetal morbidity globally, yet it is undiagnosed in 90–95% of cases. The purpose of this study was to assess precision and accuracy of a novel microsampling device for measuring cytokines in amniotic fluid (AF) to enable noninvasive evaluation of localized inflammation. Methods/Study Population: AF was obtained from discarded amniocentesis samples from 3 deidentified patients without known inflammation. Samples were spiked to 5 concentrations of interleukin-6 (IL-6) and tumor necrosis factor-a (TNF-a), then sampled using the Neoteryx Mitra volumetric microsampling device (20 µL). Dried/reconstituted samples were analyzed using the Luminex R&D Mag XL multiplex (IL-6, TNF-α) and compared to free-flowing AF. Inter-and intra-assay performances were evaluated across 5-runs in 3 standard cytokine concentrations. Recovery/linearity was assessed by a recovery curve and parameter estimates. Precision was assessed between-run and within-run using coefficients of variance (%CV). Accuracy was evaluated as agreement between microsampled and free-flowing AF using Bland–Altman plots. Results/Anticipated Results: TNF-α results were linear for all 3 patients across 5 concentrations. However, accuracy and recovery consistently failed (mean±SD percent recovery 176±21%). TNF-α results had acceptable precision with %CV within-run of 6.8–12.1% and across-runs of 14.0–15.9%. Microsampled TNF-α agreed with free-flowing sample: 16 of 18 (89%) were within 1SD of the mean difference (-24±36 pg/ml). IL-6 results were linear for 1/3 patients and had unacceptable accuracy and recovery for 13/15 samples (mean±SD percent recovery 764±469%). IL-6 results had acceptable precision (%CV within-run 6.2%, 7.5% and 11.2%; across-run .9–15.2%). Microsampled IL-6 agreed with free-flowing sample; 17 of 18 (94%) were within 1SD of the mean difference (-148±180pg/ml). Free-flowing vs. microsampling methods agreed best at low concentration. Discussion/Significance of Impact: This study provides preliminary support for noninvasive measurement of cytokines leveraging small amounts of leaking AF providing a promising alternative to amniocentesis and potential for assessing inflammation intrapartum. Clinical application requires development of reference ranges and association between cytokine levels and outcomes.

Objectives/Goals: Some children with asthma benefit from care from asthma specialists: allergist/immunologists or pulmonologists. However, a limited supply of specialists poses access problems for some children. Our objective was to describe the state of specialist care for pediatric asthma in Maine. Methods/Study Population: Using Maine all-payer claims data for April 2018–April 2021, we identified children with asthma meeting criteria for subspecialist referral based on NIH Expert Panel Report criteria. We determined which of these children had encounters with an allergist-immunologist or pediatric pulmonologist during the study period and performed logistic regressions to determine what demographic traits were associated with receipt of care. Results/Anticipated Results: Among children meeting criteria for specialists referral, 33% had an encounter with an asthma specialist. Specialist encounters were associated with younger age (p<0.001) and male sex (p = 0.003). Encounters were negatively associated with Medicaid insurance (versus commercial, p<0.001) and living in more rural areas (RUCA codes 4–10, p<0.001), lower childhood opportunity index (high or less, <0.001), and longer travel time to pediatric asthma specialists in Maine (<0.001). Discussion/Significance of Impact: Specialist care for children with complex or severe asthma is an evidence-based practice that is not equally accessible to all children in Maine. This may restrict their access to specific asthma therapies such as allergic desensitization and monoclonal antibodies.