ABSTRACT IMPACT: The implementation of nasal nitric oxide (nNO) as a diagnostic tool to understand the phenotypic/genotypic profiles of Primary Ciliary Dyskinesia (PCD) in Puerto Rico (PR) will be translated in early disease diagnosis, avoidance of comorbidities, and increase survival in our population. OBJECTIVES/GOALS: This study aims to evaluate the role of nNO levels in PCD diagnosis in the Puerto Rican population. Also, we aim to describe the clinical, genetic, and physiological characteristics of PCD in Puerto Ricans to develop a better understanding of the disease. METHODS/STUDY POPULATION: We plan to conduct a cross-sectional study on participants recruited from patients of the Pediatric Rare Lung and Asthma Institute in PR. We will compare nNO levels among genetically confirmed PCD patients, suspected PCD patients with variant of unknown significance (VUS) mutations, suspected PCD patients without genetic mutations, and age-matched healthy subjects. We plan to analyze clinical data and genetic variants to understand the natural history of the disease. The nNO measurements will be completed following previous published protocols. We will also assess the accuracy of the nNO measurements by repeating the measurements two weeks after the initial measurement. RESULTS/ANTICIPATED RESULTS: We hypothesize that many of the VUS present in our population may represent potential new founder mutations not previously reported in the literature. Our expectation is to identify new atypical PCD phenotypes contemplating the heterogenous genetic Puerto Rican pool. We anticipate that nNO levels will help to screen, identify, and confirm diagnosis of patients with clinical PCD in PR. Our findings will be translated in avoidance of further comorbidities and mortality due to earlier disease PCD diagnosis and will expand our genetic understanding about PCD in PR and other diverse populations with heterogenous genetic admixture. DISCUSSION/SIGNIFICANCE OF FINDINGS: We present a significant and novel research proposal that plan to impact the quality of life of patients living with PCD in PR. The implementation of state-of-the-art diagnostic tools like nNO measurement will positively impact and expand our current capabilities to diagnose rare lung diseases like PCD on the island.

OBJECTIVES/GOALS: The purpose of this study is to understand how hemodynamics during dialysis in End-Stage Renal Disease (ESRD) patients on hemodialysis (HD) affect white matter health and how those effects cause cognitive impairments. METHODS/STUDY POPULATION: We collected demographic data, comorbidities, intradialytic measurements of blood pressure and cerebral oximetry, cognitive measures in several domains using NIH Toolbox Cognition Battery, diffusion-weighted and anatomical MRIs for 20 participants on HD. Specific tracts were identified using tractography and were used to calculate the average DTI measurements in each tract. Regression analysis was used to examine the relationship between mean DTI measurements of white matter integrity and cognitive performance scores. In addition, we compared diffusion MRI and T1 anatomical images of 16 healthy age-matched controls from a previous study. RESULTS/ANTICIPATED RESULTS: In our cohort 18 participants had imaging data that could be used in the analysis. We found widespread decreases in DTI white matter integrity compared to healthy age-matched controls, mean whole-brain fractional anisotropy was .3218 in the HD cohort and .3472 in controls p = .0018. Decreased integrity was found in most of the tracts identified but more decreased in tracts implicated in cognition. Partial regression analysis identified significant relationships between the white matter integrity of the left superior longitudinal fasciculus and overall fluid cognitive performance, R = .5525, p = .0174 before multiple comparisons correction when controlling for differences due to age. DISCUSSION/SIGNIFICANCE OF IMPACT: We found a widespread decrease in white matter integrity and significant correlations between cognitive performance and specific tract integrity in our HD cohort using regions identified by tractography imaging analysis. This analysis shows that HD patients have decreased white matter health and identifies several tracts that are important for cognitive performance in HD patients.

Objectives/Goals: RF-DASH trains first responders with agricultural health and safety knowledge to prevent farm accidents and improve emergency preparedness in rural communities. We used a collaborative instructional design approach to develop an online version of this in-person, in-demand, highly effective training to increase its reach and enhance its impact. Methods/Study Population: The RF-DASH project team engaged e-learning specialists to develop learning objectives and outcomes, establish course structure, curate elements of the RF-DASH in-person curriculum into e-learning materials, and design interactive course content. We then conducted user testing including an initial content review by expert RF-DASH trainers, two rounds of user testing with agricultural safety professionals and first responders. Feedback from user testing was integrated into the final version of the online training and helped inform the evaluation plan that was developed prior to the online training launch in October 2025. Results/Anticipated Results: The RF-DASH Online Course uses interactive learning techniques, including 360° virtual tours of actual farms, animated videos, and knowledge checks. The virtual tours were deemed critical by reviewers for translating the in-person training to an online format while maintaining the real-life applicability that makes the intervention effective and appealing. The co-design approach required flexibility and humility among all actors, which resulted in a stronger product and deep learning for all. The opportunity to “learn what you don’t know you don’t know” was crucial and is not always supported by funders, timelines, or team capacity. For translational science to achieve the breadth and depth of its potential for impact, these challenges should be met with support rather than reluctance whenever possible. Discussion/Significance of Impact: The collaborative co-design process used to create the online version of the training was critical to the quality and relevance of the final product, and the learning that resulted on behalf of the RF-DASH team speaks to the utility of conducting a needs assessment at the outset of adapting any intervention for new audiences and aims.

OBJECTIVES/GOALS: The objective of this study is to examine the independent and combined effectiveness of a multiple micronutrient supplementation and responsive caregiving intervention on early child development in southwestern Guatemala. METHODS/STUDY POPULATION: We conducted a double blind, 2 x 2, cluster randomized controlled trial combining micronutrient supplementation and responsive caregiving treatments. We enrolled 309 infants (6-18 months) and 387 preschoolers (36-52 months) at nutritional risk (height-for-age z-score < -1) (51% male; 17% indigenous ethnicity; 76% of caregivers completed ≤ primary school). The supplementation arm received a maize-soy product fortified with 21 micronutrients for 6 months; the control was fortified with B2. The responsive caregiving intervention was adapted from the UNICEF Care for Child Development program and delivered over 6 home visits. We examined changes in early child development from baseline (2015) to endline (2017) using the Bayley Scales of Infant and Toddler Development III and Bracken Scales of School Readiness. RESULTS/ANTICIPATED RESULTS: Among infants, age standardized Bayley cognitive development scores declined over time in each treatment arm. Infant Bayley motor development scores significantly increased in the combined multiple micronutrient supplementation + responsive caregiving arm (â^† = 3.67 [95% CI: 0.17, 7.17], p = 0.04). Among preschoolers, school readiness decreased over time in each treatment arm. The rates of change in infant development and preschooler school readiness did not differ between treatment arms. Further analysis will explore effect modification of the intervention by pre-specified child and household factors. DISCUSSION/SIGNIFICANCE: Combined multiple micronutrient supplementation and responsive caregiving supported motor development while cognitive and school readiness were not impacted by the intervention. These findings will inform the effectiveness and beneficiaries of multisectoral interventions to promote early child development in adverse environments.

OBJECTIVES/GOALS: The global incidence of calcific aortic valve disease (CAVD) increased 3.5-fold since 1990. No preventative or therapeutic pharmaceutical therapies exist for CAVD. We will establish the therapeutic potential of osteoclast-derived exosomes though characterization of contents and mechanisms of action to protect against mineralization. METHODS/STUDY POPULATION: Exosomes were purified from conditioned media collected from murine myeloid precursor cells, RAW264.7 (control), and osteoclasts induced to differentiate from RAW264.7 cells (OD). Protein content of exosomes was determined using proteomic analyses. Nucleic acid contents will be identified by sequencing mRNA, miRNA, and DNA. The calcification prevention and reabsorption abilities of control and OD exosomes will be tested using human valvular interstitial cells (VIC) and smooth muscle cell calcification assays and acellular osteologic disc assays, respectively. Comparison between cellular and acellular systems will help identify mechanisms of action, and demonstrate potential therapeutic viability of OD exosomes in preventative vs resorptive treatments. RESULTS/ANTICIPATED RESULTS: OD exosomes, but not control exosomes, prevented calcification in VIC in vitro. OD exosomes contained osteoclast-specific proteins including TRAP, MMP6, cathepsin K, and bone reabsorption factors including V type proton pumps, ATPases, and integrins. These genes are also involved in resorptive activities, and were highly upregulated in OD compared to control exosomes. We anticipate miRNA signatures associated with mineral resorption will also be present. Increased knowledge of exosome cargo will illuminate their mechanism of action and allow future work to engineer increased efficacy. We also anticipate a therapeutic response when OD exosomes are applied after calcification has begun, showing exosomes promote calcium reabsorption. DISCUSSION/SIGNIFICANCE: Establishing therapeutic potential and examining mechanisms of action will pave the way for OD exosomes as a CAVD treatment. Analysis of exosome contents will determine active molecules to be enhanced in future studies. This work will lay a foundation for moving into aortic valve organoid models, which are accepted by the FDA for preclinical trials.

OBJECTIVES/GOALS: There is a need for high-quality and efficient translation of health technologies in the Ontario healthcare system. The goal of this project is to understand the decision-making processes of government expert groups developing recommendations for the system-level implementation of pharmacogenomic testing. METHODS/STUDY POPULATION: This prospective observational case study includes the Ontario Health Pharmacogenomics (PGx) Working Group focused on developing recommendations for a PGx testing implementation strategy in the province. Ontario Health is the government agency that oversees provincial healthcare planning and service delivery. Using qualitative ethnographic methods, we will observe and document the working group’s activities over a 10-month period. Data collection involves meeting recordings, correspondences, researcher field notes, decision-making processes, and group characteristics. Using descriptive statistics and inductive qualitative analyses, the data will be examined to build theory and frameworks for knowledge translation. RESULTS/ANTICIPATED RESULTS: The results will be presented through a case report, process maps, decision milestones, visualizations, and procedural recommendations for future expert groups. This study will contribute to the body of foundational knowledge about translational sciences and support the National Center for Advancing Translational Sciences’guiding principles. To enhance translational processes and train the future translational workforce, this research can be used for educational initiatives. In addition, the observed processes will inform a theory about how expert recommendations are developed in public healthcare systems. DISCUSSION/SIGNIFICANCE: This research addresses a current gap in understanding around translational processes, government decision-making, and the development of recommendations for the adoption, implementation, and dissemination of the novel health technologies transforming public healthcare in Canada.

OBJECTIVES/GOALS: The purpose of this secondary data analysis was to identify latent subgroups of seriously ill adults based on multiple chronic conditions and mortality risk using the CCI. This study was conducted by performing a secondary analysis of data from a randomized controlled trial of seriously ill patients receiving palliative care. METHODS/STUDY POPULATION: A cross-sectional analysis of baseline CCI data was conducted. 381 seriously ill adults receiving palliative care were in the original study. Latent subgroups were identified based on the CCI by conducting a latent class analysis in MPlus. The LCA was modeled on each of the 19 disease items as binary latent predictor variables, an additional binary variable representing presence of any disease not accounted for by the CCI, and a final categorical variable representing the total CCI score divided based on clinically significant cutoffs including zero, low (> = 1-<2), moderate (> = 2-<5), and high CCI (> = 5). RESULTS/ANTICIPATED RESULTS: Three distinct latent subgroups were identified based on the CCI. Latent subgroup 1 included those with a low-moderate CCI consisting of MCC and non-Metastatic Cancers (n = 178), with 45% of this group having chronic obstructive pulmonary disease. The second two subgroups included individuals with a high CCI or a score greater than or equal to 5. Latent subgroup 2 (n = 64) was comprised of individuals with MCC and non-metastatic cancer. Latent subgroup 3 (n = 139) included individuals with metastatic cancer. DISCUSSION/SIGNIFICANCE OF IMPACT: In a sample of seriously ill adults with MCC, latent subgroups were identified consisting of individuals with low, moderate, or high CCI. The low to moderate CCI group consists of individuals with chronic conditions including COPD, congestive heart failure, myocardial infarction, cardiovascular disease. There were two subgroups with high CCI scores and the differentiating factor between the two subgroups was the presence of metastatic cancer in latent subgroup 3. The identification of latent subgroups sets the groundwork for further analyses to compare differences in symptom burden, quality of life, and functional status between groups. The findings have the potential to inform future studies seeking to better characterize seriously adults with MCC based on their disease burden and mortality risk.

OBJECTIVES/GOALS: The Fulkerson Home Food Inventory (HFI) is widely used to assess the home food environment, a key target of behavioral weight loss trials. However, no standardized report is available. We created publicly available procedures to automate and standardize HFI reporting, yielding a personalized report to enhance this measures clinical utility. METHODS/STUDY POPULATION: Parents in the TEENS adolescent behavioral weight loss trial complete the HFI at 0-, 2-, 4-, 8-, and 12m and receive personalized reports at each timepoint. In REDCap, participants identify foods available in their home. HFI syntax is applied to calculate the obesogenic home food availability score. Categories of foods found are identified, with specific guidance provided to enhance their home food environment. Prior to automation, procedures were time intensive and error prone. To address this, HFI data are exported into Excel by a PowerShell (v7.2) command-line script using Python (v3.10) with the REDCap API. Results are calculated with F# (v6.0) using Microsoft Excel Interop API and inserted into a report template with F# using the Microsoft Publisher Interop API. This process is repeated at each timepoint. RESULTS/ANTICIPATED RESULTS: The new automated procedures significantly reduce time to generate reports and enhance accuracy. Procedures yield a 2-page individualized report that includes the obesogenic home food environment score and identifies categories of healthy items found (e.g., fruits, vegetables, whole grains) as well as areas of improvement (e.g., high-fat dairy products, processed meats). Specific items found in each category are identified. The report identifies food found in the home (e.g., chicken nuggets) with suggested healthier substitutions (e.g., lean chicken breast). This syntax and commands will be made publicly available for use in the scientific and clinical community. DISCUSSION/SIGNIFICANCE: These publicly available procedures optimize, automate, and standardize reporting for the HFI. Procedures improve efficiency within large-scale clinical trials and yield a personalized report to enhance the clinical utility of this measure and empower participants to make informed decisions about their health behaviors.

Objectives/Goals: Community gardening can foster healthy behaviors among low-income communities. This project aimed to develop a community garden. The primary objectives of this project are (1) assessing the need for and perspective on a community garden at the childcare center, (2) installing the garden, and (3) engaging children in gardening education. Methods/Study Population: This project took place at a childcare center in Harrisburg, PA. Most (74.6%) residents identified as Black or Hispanic/Latino. Every child at the center was eligible for free or reduced lunch. A listening session was held with directly impacted community members to discuss the need for a community garden. Four caregivers, 1 early childhood educator and a master gardener (n = 6) attended the listening session, in which they shared their personal strengths and challenges in growing food. Attendees provided suggestions on what foods they wanted to grow. Children enrolled in the center’s summer program (n = 50) were then invited to participate in weekly gardening activities for 9 weeks. Activities were targeted to preschoolers (3- to 5-year-olds). Older children enrolled in the summer program were welcome to participate. Results/Anticipated Results: Feedback from the listening session was positive. Attendees provided ideas on what to grow and shared interest in expanding the garden to the broader community. Project staff installed four garden beds and planted a variety of herbs (basil, mint, and lavender), fruits (strawberry and melon), and vegetables (tomato, squash, pepper, and onion). Roughly 20–50 children were engaged in the garden each week. Eight weeks into the project, one member from the broader community noticed the garden’s growth and expressed gratitude to the staff, stating “I saw you when you first started planting. This is great what you are doing for the kids.” Children and the center’s staff responded positively to the activities. The staff expressed verbal gratitude for the project and were enthusiastic about maintaining the garden. Discussion/Significance of Impact: Developing a community garden was feasible in this sample and shows potential to (1) increase children’s food literacy and vegetable acceptance and (2) bridge the gap from farm to early childcare education. The project’s success paves way for future gardening initiatives that address food access issues within other diverse low-income populations.

OBJECTIVES/GOALS: The overall objective of this study is to determine if medial arterial calcification (MAC) is an independent predictor of diabetes and to evaluate the relationship between MAC, periodontitis and Type II diabetes mellitus. METHODS/STUDY POPULATION: A retrospective case-control model analyzing radiographs for periodontitis and MAC to identify potential biomarkers for underlying systemic conditions, such as diabetes. Charts of patients attending UPENN School of Dental Medicine clinics between 2015 and 2022 were reviewed. Demographics, medical and dental history, diabetic status (identified by POC blood glucose level, fasting blood glucose and/or A1C), and medication history were documented amongst other variables. Patients aged 18 years or older with diabetes and having full mouth intraoral radiographs (FMX), panoramic radiographs and CBCTs were included. Patients with radiographs of poor quality were excluded. Multivariate analysis was used to determine possible associations between diabetes and periodontitis among persons with or without MAC. RESULTS/ANTICIPATED RESULTS: In our pilot study involving 28 participants, 53% of the population with moderate or severe periodontitis had MAC. By the Fisher’s Exact Test, there was an association, meaning those with more periodontal disease are more likely to have MAC (p=0.014). Sixty-three percent of patients with diabetes had MAC, while 19% of patients without evidence of diabetes also had MAC, (p=0.067). There was not enough evidence of association between diabetes and presence of MAC at this time, due to a small sample size, however there was a high prevalence of MAC among the diabetics. We hypothesized that periodontitis, a condition that shares many risk factors with diabetes would also be associated with incident MAC. Findings from this study will be key for the implementation of preventive screening protocols and referral systems. DISCUSSION/SIGNIFICANCE: Diabetes is on the rise and about half of diabetics are undiagnosed. CBCT imaging frequently used in dentistry can detect incidental findings such as MAC. This study has the potential of detecting statistically significant links between MAC, periodontitis and diabetes, hence serving as a sensitive radiographic biomarker for diabetes.

OBJECTIVES/GOALS: eHealth programs centered on cognitive behavior therapy (CBT) can be supported by physical therapists to feasibly deliver psychologically-informed physical therapy (PIPT). This study assessed patient and physical therapist (PT) perspectives of adding a CBT-based eHealth program to physical therapy. METHODS/STUDY POPULATION: In our uncontrolled pilot study, PTs were trained in motivational interviewing (MI) to support patient engagement with a 7-module eHealth CBT-based pain self-management program that accompanied a course of PT. Interviews were conducted with a convenience sample of 13 patients with chronic back and/or neck pain and 9 PTs to evaluate experiences with the eHealth program, perceived benefits, barriers and facilitators to integration, and future recommendations for implementation from both perspectives. Interview data were recorded, transcribed, and analyzed using qualitative content analysis for core themes. RESULTS/ANTICIPATED RESULTS: Patients benefited from the eHealth program, especially relaxation (69% of respondents) and meditation/mindfulness (62%). Time and technology were barriers for eHealth engagement (23% and 15%). Most (92%) patients reported their therapist reviewed eHealth progress, although one-third of patients wanted more discussion and feedback on eHealth content. PTs felt the eHealth program was valuable (67%), user-friendly (56%), and took minimal effort to support (78%). Some (44%) therapists felt more confident after MI training to support patient engagement. PTs recommended offering patients tablets (i.e., iPads) for clinic or home use and additional therapist training in MI. DISCUSSION/SIGNIFICANCE: PTs and patients valued a clinician-supported eHealth program, suggesting an acceptable option for PIPT. Future implementation work aims to identify optimal therapist training in MI and additional strategies to overcome time or technology barriers.

OBJECTIVES/GOALS: Residents of environmental justice (EJ) communities experience significantly higher rates of negative health outcomes associated with poor air quality. Low-cost air sensors may supplement regulatory monitoring to better measure air pollution at local scales, but widespread application of this technology remains limited due to many challenges. METHODS/STUDY POPULATION: To address these obstacles, we designed a training program to equip community and academic research partners with the skills and knowledge to successfully apply low-cost sensors in community-engaged environmental health research. The R esearch Innovations using Sensor Technology in Environmental Justice Communities (RISE Communities) program was established through an NIEHS R25 award in 2022 and has three specific aims: 1) Foster community-academic partnerships through research education, training, and team development activities, 2) Provide technical training in the application of low-cost sensors for indoor, outdoor, and personal air monitoring in EJ communities, and 3) Establish a community of practice to address air quality in communities nationwide. RESULTS/ANTICIPATED RESULTS: We hosted our first cohort in August 2023, training five community-academic research teams in team collaboration, community-engaged research, and technical skills for collecting and analyzing data from PurpleAir sensors. Each team received 12 sensors to take to their home EJ communities to begin their projects. Community of practice development continues through interactive webinars and development of a web-based repository of training videos and discussion board posts. Evaluation data show high participant learning and satisfaction, with mean confidence scores improving on 6/8 metrics. Evaluation data also suggest several areas for improvement such as more time spent in teams for planning and additional opportunities for interaction within the cohort and with program instructors for problem-solving. DISCUSSION/SIGNIFICANCE: Effective training for team-based community-engaged research requires careful planning for team development and study implementation. Longitudinal training and support for the technical aspects of utilizing air sensors is also critical to team success. The RISE Communities program is actively recruiting for future training cohorts.

Objectives/Goals: As the aging population increases, maintaining cognitive and physical health becomes crucial. Executive functions (EF), including reaction time, sustained attention, and spatial memory, are essential for daily life and independence in older adults but tend to decline with age, especially in Alzheimer’s disease (AD) patients. Methods/Study Population: Physical balance (PB) impairments further exacerbate this decline, affecting the quality of life and independence. Balance training (BT) emerges as a potential non-pharmacological intervention to enhance EF. This study aims to 1) assess the association between PB impairments and EF deterioration in older adults with AD; 2) evaluate the impact of BT on EF enhancement in this population; and 3) explore changes in brain-derived neurotrophic factor (BDNF) biomarkers before and after physical activity related to BT. This pilot study employs a cross-sectional design with older Hispanic adults (>65 years) in Puerto Rico, comparing a control group without BT intervention to an experimental group with BT intervention. Both groups will be assessed for EF and BDNF biomarkers pre- and post-physical activity. Results/Anticipated Results: Expected outcomes include identifying significant correlations between PB impairment and diminished EF, demonstrating measurable EF improvements following BT, and evidencing sustained BDNF release post-BT despite PB impairments. By understanding the biological mechanisms linking BT and cognitive improvements, particularly the role of BDNF, this research could inform future strategies to mitigate cognitive decline in AD patients through targeted physical interventions. Discussion/Significance of Impact: The findings of this study could provide valuable insights into the development of new preventive interventions, enhancing the quality of life and independence for older adults.

Objectives/Goals: We aim to enhance risk prediction in kidney transplantation outcomes by improving models of peptide antigen presentation of mismatched HLA molecules. HLA-derived peptides presented by recipient HLA Class II to T-cells can activate an immune response ultimately leading to graft failure. Methods/Study Population: T-cell epitope models for HLA mismatching struggle to predict which peptides are presented because antigen processing by proteases is not well modeled. We model antigen processing of HLA Class II proteins using 3D HLA structures (crystallography data) to create an HLA-specific Antigen Processing Likelihood (APL) model. APL uses conformational stability measurements to predict cleavage sites from proteolysis. We integrated APL into a T-cell epitope prediction tool for HLA-derived peptides called PIRCHE-II that is currently used to risk-stratify risk of kidney allograft failure based on donor and recipient HLA genotypes. To measure risk stratification of APL-informed peptide predictions, we will use a historical kidney transplant cohort from 2000 to 2023. Results/Anticipated Results: We expect the application of APL could reduce false- positive peptide binders that influence risk prediction scores. We anticipate improved peptide prediction accuracy compared to existing tools such as NetMHCIIPan which assumes all possible peptides are equally likely to emerge from antigen processing. NetMHCIIPan is currently used by PIRCHE-II HLA mismatch risk algorithm. Out of 12 donor–recipient transplant pairs, NetMHCIIPan has found an average of 41 peptides per pairing and APL found an average of 62 peptides per pairing. These peptides are unique to each prediction, so a combined prediction could reduce the peptide list. We expect that merging antigen processing (APL) and peptide binding (NetMHCIIPan) models into a unified model would enhance risk stratification for graft failure compared to PIRCHE-II. Discussion/Significance of Impact: More holistic modeling of immune pathways can lead to more realistic numbers of peptides found from mismatched HLA proteins. Understanding how HLA matching contributes to kidney transplant outcomes can better stratify risks for recipients, enabling personalized treatment to induce immune tolerance and ultimately improving outcomes.

ABSTRACT IMPACT: This study deepens knowledge with respect to the associations between depression, cardiometabolic conditions, and accelerated aging with a clinically accessible marker in a population with disproportionate risk for comorbidity. OBJECTIVES/GOALS: The aim of this secondary analysis is to examine associations between DNA methylation age acceleration (DNAm AA) and depressive symptoms in African American women (AAW) considering the presence of cardiometabolic conditions (CMCs) including hypertension, diabetes, obesity. METHODS/STUDY POPULATION: Genomic and longitudinal clinical data (collected 2015-2020) from the Intergenerational Impact of Genetic and Psychosocial Factors on Blood Pressure Study (InterGEN) cohort (n=227) were utilized for this analysis. DNA methylation age (estimated by the Horvath method) incorporates DNA methylation status at 353 CpG sites. DNAm AA is the residual of DNA methylation age regressed on chronological age in a linear model. Spearman’s correlations and linear regression examine the relationship between DNAm AA and depressive symptoms (Beck Depression Inventory) and cardiometabolic status. The potential association and impact of SES, trauma, substance use, and stress were also considered. RESULTS/ANTICIPATED RESULTS: Contrary to our hypothesis, DNAm AA did not associate with the severity of depressive symptoms. Correlation between DNAm AA and affective symptom subscore (BDI) approached significance (p = 0.06). We observed significant correlations between DNAm AA and specific depressive symptoms including participants’ reported disappointment, disgust, or hatred toward themselves (p < 0.05), difficulty with making decisions (p < 0.05), and worry about their physical health (p < 0.05). DNAm AA was also significantly correlated with BMI (p > 0.001). Significant relationships were not evident in the subsequent regression analysis examining potential relationships between DNAm AA and depression. To our knowledge, this is the first study to examine associations between DNAm AA and depressive symptoms in AAW. DISCUSSION/SIGNIFICANCE OF FINDINGS: Depression limits life quality and quantity and is highly comorbid in CMCs. AAW have a high risk of comorbidity. This study deepens knowledge with respect to the associations between depression, CMCs, and aging with a clinically accessible marker in a population with disproportionate risk.

OBJECTIVES/SPECIFIC AIMS: Our objective was to examine serum E2 levels in dcSSc males in relation to disease characteristics (i.e autoanitbody profile and internal organ involvement) and its impact on survival. METHODS/STUDY POPULATION: We measured serum E2 levels in 83 dcSSc men >50 years old from the University of Pittsburgh Scleroderma Center and healthy controls of similar age. Using statistical modeling, we examined the associations between circulating E2 levels, internal organ involvement, autoantibody profiles, and survival. RESULTS/ANTICIPATED RESULTS: Male dcSSc patients had significantly higher serum E2 levels compared to healthy male controls and compared to dcSSc post-menopausal women of similar age. Male dcSSc patients with high serum E2 levels had significantly more heart involvement and worse survival. Using Cox regression modeling for risk of death, increasing serum E2 levels in anti-Scl-70 antibody positive dcSSc males were associated an increased risk of death. DISCUSSION/SIGNIFICANCE OF IMPACT: DcSSc male patients have higher levels of E2 compared to healthy controls and dcSSc postmenopausal women. Elevated serum E2 levels in dcSSc males >50 are associated with heart involvement and, if anti-Scl-70 antibody positive, worse survival. Our current study expands on our previous work, not only that that E2 exerts pro-fibrotic effects on skin, but also internal organ involvement, overall survival. These data suggest an important role of estrogen imbalance in SSc.

OBJECTIVES/GOALS: Childhood psychiatric symptoms are highly comorbid. Their co-occurrence and association with negative life outcomes is partially explained by deficits in executive control, or processes enabling self-regulation. Here, we test a novel executive neural target in three fMRI tasks and its relevance to shared psychopathology. METHODS/STUDY POPULATION: We studied 60 children [15 F/45 M; mean age (SD)=11.6 years (1.62)] with diverse diagnoses including attention deficit disorder (n=26) and autism spectrum disorder (n=22). We extracted a latent general factor of psychopathology using principal component analyses applied to parent-report Child Behavior Checklist syndrome scores. Subjects completed 3 executive control fMRI probes, tapping adaptive control, working memory, and inhibition. Correlational psychophysiological interaction (cPPI) analysis measured correlations between executive control-related modulations of activity in 414 network-affiliated parcels. We selected parcels exhibiting control-related cross-network correlations as well as control-related activity across all tasks and tested them for association with psychopathology. RESULTS/ANTICIPATED RESULTS: cPPI connectivity matrices were thresholded and graphs were identified using the Network-Based Statistic toolbox (p90th percentile PC) as well as control-related activation (>10% activated voxels; p DISCUSSION/SIGNIFICANCE: Our results examine cross-network interactions between brain regions during 3 fMRI tasks and their role in explaining individual variation in psychopathology. As executive control links to both comorbidity and life outcomes, identifying the clinically-relevant neural correlates of controlled behavior may lead to transdiagnostic treatments.