Objectives/Goals: To benchmark the extent to which variability in automated WMH quantification influences stroke and dementia risk stratification by characterizing differences in volume estimates and spatial concordance across three segmentation tools (BIANCA, SAMSEG, and nnU-Net). Methods/Study Population: This retrospective, cross-sectional study will analyze de-identified clinical brain MRI scans from patients aged ≥50 years, including those with and without prior stroke, obtained from institutional imaging repositories. Eligible scans include co-registered T1-weighted and FLAIR sequences. Scans with poor image quality and scans from patients with multiple sclerosis, other demyelinating disorders, or brain tumors will be excluded. WMHs will be segmented using three tools: BIANCA (supervised k-NN classifier), SAMSEG (unsupervised Bayesian model), and nnU-Net (self-configuring deep learning). Agreement in WMH burden will be assessed with intraclass correlation coefficients (ICC[3,1]) and spatial concordance with Dice similarity coefficients (DSC). Bland-Altman analyses will evaluate bias. Results/Anticipated Results: We will analyze 600 brain MRI scans. All three tools are expected to detect similar WMH spatial distributions, though total WMH volume estimates may differ. nnU-Net may yield higher WMH burden estimates than BIANCA or SAMSEG. WMH burden will be neuroradiologist-adjudicated to establish the gold standard, against which concordance for each tool and inter-tool agreement will be assessed using ICC[3,1] and DSC. We anticipate moderate-to-high agreement, but tool-specific variability could influence reproducibility and interpretation of WMH burden for future WMH-related research studies. This study will quantify the extent of variability among tools in clinical brain MRI scans and evaluate whether these differences could meaningfully alter future stroke risk classification thresholds. Discussion/Significance of Impact: This study will benchmark automated segmentation tools for WMH-related research by standardizing WMH quantification to improve reproducibility and reliability for stroke and dementia risk stratification. Establishing tool-specific benchmarks may also guide integration of automated WMH measures into clinical decision-support systems.

Objectives/Goals: In mice, it has been shown that loss of Cib2 (calcium and integrin-binding protein 2) results in progressive retinal disease that recapitulates many characteristics of age-related macular degeneration (AMD). This study aims to characterize transcriptional changes in the retinal pigment epithelium (RPE) that underlie this disease process. Methods/Study Population: RPE tissue samples, pooled from 2–3 mice for each biological group, were collected from Cib2-KO and wildtype (WT) mice at two (young) and eight (aged) months of age. Bulk mRNA sequencing was performed using the Illumina HiSeq 4000. Reads were aligned to the UCSC mouse reference genome and quantified using HTSeq. Significant differentially expressed genes (DEGs) between mouse genotype and age groups were assessed using DESeq. CLICK unsupervised clustering followed by gene ontology analysis was performed to identify cellular processes and molecular pathways affected by loss of Cib2 as well as age. Results/Anticipated Results: CLICK analysis revealed several functional pathways that are differentially expressed between sample groups. For example, in both young and aged mice, pathways upregulated in Cib2-KO samples included calcium signaling, RhoA signaling, and integrin signaling. Uniquely downregulated DEGs in young Cib2-KO animals were related to complement and coagulation cascades, LXR/RXR activation (related to lipid synthesis and transport), and phagosomes. Aged Cib2-KO mice displayed the most significant downregulation of genes in the phototransduction pathway, indicating temporal changes in functional pathways that correlate with disease progression. Next steps in analysis include investigating patterns in RPE- and AMD-signature gene sets that may identify molecular pathways more specific to human disease. Discussion/Significance of Impact: Many current studies investigate the role of complement activation, vesicle trafficking, and ion transport as top contributors to AMD development. We identified DEGs paralleling many of these molecular pathways in Cib2-KO mice, highlighting their potential as a model to study age-related RPE pathologies and evaluate therapeutic interventions.

OBJECTIVES/GOALS: Studying IPF associated health care utilization (HRU) in real world settings, provides the opportunity to produce generalizable results that can directly inform models of care delivery. The objective of this study was to examine real-world differences in the natural history of annual HRU and treatment trends associated with IPF in a large, community-based population of patients with IPF, compared to matched non-IPF controls. METHODS/STUDY POPULATION: Cases of IPF were identified using case validated algorithms in the Kaiser Permanente Northern California adult population from 2000 to 2014. Each case was matched to at least one and no more than five non-IPF controls by age, sex, race/ethnicity and length of enrollment. The date of the first occurrence of the IPF-specific diagnostic code was considered the index date for cases and matched controls. Comorbidity burden and HRU was assessed in the five years pre- and post-index date, including hospitalizations, outpatient visits, use of diagnostic and monitoring studies and medications. Poisson generalized estimating equations models with robust standard errors were used to estimate adjusted case-control differences in HRU, accounting for clustering within matched sets. RESULTS/ANTICIPATED RESULTS: 691 patients were identified with incident IPF during the study period and matched to 3,452 control subjects. Demographics were well balanced between cases and controls due to matching. Patients with IPF had a higher burden of all selected co-morbidities and higher HRU compared to controls. In the pre-index period, IPF members had significantly higher rates of all diagnostic procedures (p < 0.001) and health care visits, with the exception of skilled nursing facility care (p < 0.001). The greatest relative difference was observed with use of Chest CT (RR = 245.94, 95% CI 117.04, 516.82). In the post-index period compared to controls, patients with IPF had higher rates of serial testing (p < 0.001) and inpatient and outpatient care including, all-cause hospitalization (1.55), emergency room visits (1.19), outpatient visits (1.18), and skilled nursing facility stay (1.35). DISCUSSION/SIGNIFICANCE OF IMPACT: Patients with idiopathic pulmonary fibrosis experience increased co-morbidity and healthcare resource utilization compared to controls. This increased burden extends beyond the index-period as previously documented and is true for a large real-world cohort. CONFLICT OF INTEREST DESCRIPTION: NA

ABSTRACT IMPACT: By demonstrating the feasibility of a multimodal, interdisciplinary intervention for prediabetes, the current project aims to provide a template for the prevention of diabetes and associate comorbid conditions. OBJECTIVES/GOALS: To determine if a multi-modal, interdisciplinary intervention delivered to a group of prediabetic patients will result in reduced rates of diabetes progression. This project is a retrospective evaluation that will exam the feasibility and possibly efficacy of this intervention. METHODS/STUDY POPULATION: We will enroll 50 participants for the clinic, aged 21-60 inclusive. Patients will have a Body Mass Index >27kg/m2 with a diagnosis of prediabetes. Patients must be non-pregnant, using approved contraception, and agree to not become pregnant for 1 year after enrollment. After enrollment, the initial treatment period is for 1 year and includes a 12 week low calorie diet plan, a 6-month intensive behavioral and lifestyle modification plan followed by a 6 month behavior reinforcement extension. Weight management medications may be used if appropriate for the patient from a clinical perspective during the 6-month intensive behavioral/lifestyle modification. RESULTS/ANTICIPATED RESULTS: It is anticipated that there will be decreased weight with a mean weight loss goal of approximately >10%. Furthermore, it is expect that there will be improvement of other markers of metabolic disease. These include improvement of lipid values (LDL-C, HDL-C, Triglycerides, Total Cholesterol) as well as blood pressure with expected blood pressures of below 130/80 in greater than 50% of participants. Finally, It is expected that 50% or greater participants will have improvement of glycemic control. It is anticipated that greater than 50% of participants will have improvement of glycemic control and achieve normoglycemia. These values will be determined based upon fasting glucose or A1c. DISCUSSION/SIGNIFICANCE OF FINDINGS: The significance of this intervention is enormous. By demonstrating feasibility in this trial, we can work toward both assessing efficacy and possibly dissemination of this model program. If these interventions provide durable changes at scale, this could help slow the epidemic of obesity and obesity related comorbid conditions.

OBJECTIVES/SPECIFIC AIMS: The study aims to further investigate how cyclophilin D (CypD), the key mPTP opening regulator, affects BMSCs fate and to determine potential regulatory mechanisms involved in CypD regulation during osteogenesis. METHODS/STUDY POPULATION: We evaluated CypD mRNA expression in mouse BMSCs and in osteogenic-like (OL) cells during the course of OB differentiation. CypD protein level was also probed. Moreover, BMSCs had their mPTP activity recorded during osteoinduction. We further analyzed the effect of CypD genetic deletion on osteogenesis in vitro and in vivo. For our in vivo model, we performed the ectopic bone formation assay to asses differences in ossicle formation when CypD KO BMSCs were transplanted compared to wild type littermate BMSCs. In our in vitro model, we transfected OL cells with either CypD gain of function or CypD loss of function vector and measured their osteogenic differentiation potential. Additionally, we treated BMSCs with CypD inhibitor and compare to non-treated BMSCs for mineralization level. To determine potential regulatory mechanisms involved in CypD regulation, we analyzed the CypD gene (Ppif) promoter for potential transcription factor (TF) binding sites and found multiple Smad-binding elements within this promoter. Smads (Smad1, 5, 8) are TFs downstream from Bone Morphogenic Protein (BMP) signaling pathway that transmit cell differentiation signaling, and exert either activating or inhibitory effects on a variety of genes. We also transfect OL cells with Smad1 vector and analyzed for CypD mRNA levels. RESULTS/ANTICIPATED RESULTS: - Our data showed that CypD mRNA levels decreased in both primary cells and OL cells at day 7 and day 14 in osteogenic media. - Osteogenic induction also decreased mPTP activity. - In vivo ectopic bone formation assay showed increased ossicle fo DISCUSSION/SIGNIFICANCE OF IMPACT: Our data suggest that downregulation of CypD increases OB differentiation due to improved OxPhos activity led by mPTP closure. Our results corroborate reports of CypD downregulation and mPTP closure during neuronal differentiation in developing rat brains as well as in cardiomyocyte differentiation in developing mouse hearts. Our studies also suggest a yet unknown mechanism linking differentiation signaling with mitochondrial function – BMP/Smad mediated downregulation of CypD transcription. As initially mentioned, in a previous study, our lab showed that CypD KO mice present higher mitochondrial function and osteogenicity in aged BMSCs and less osteoporosis burden. Taken together, these results suggest that CypD can be a potential target to prevent bone loss in aging.

OBJECTIVES/SPECIFIC AIMS: We aimed to develop an assay to measure new protein synthesis after Antisense Oligonucleotide treatment, which we hypothesized to be the earliest biochemical identification of RNA-targeting therapy efficacy. METHODS/STUDY POPULATION: We treated 2 transgenic animal models expressing proteins implicated in neurodegenerative disease: human tau protein (hTau) and human superoxide dismutase 1 (hSOD1), with ASO against these mRNA transcripts. Animals received isotope-labeled 13C6-Leucine via drinking water to label newly synthesized proteins. We assayed target protein synthesis and concentration after ASO treatment to determine the earliest identification of ASO target engagement. RESULTS/ANTICIPATED RESULTS: hTau ASO treatment in transgenic mice lowered hTau protein concentration 23 days post-treatment in cortex (95% CI: 0.05%–64.0% reduction). In the same tissue, we observed lowering of hTau protein synthesis as early as 13 days (95% CI: 29.4%–123%). In hSOD1 transgenic rats, we observed lowering of 13C6-leucine-labeled hSOD1 in the cerebrospinal fluid 30 days after ASO treatment compared with inactive ASO control (95% CI: 12.0%–48.4%). DISCUSSION/SIGNIFICANCE OF IMPACT: In progressive neurodegenerative diseases, it is crucial to develop measurements that identify treatment efficacy early to improve patient outcomes. These data support the use of stable isotope labeling of amino acids to measure new protein synthesis as an early pharmacodynamics measurement for therapies that target RNA and inhibit the translation of proteins.

OBJECTIVES/GOALS: The Building Up Study tests the effectiveness of an intervention aimed to diversify the workforce using a two-arm cluster randomized trial. We examined how underrepresented (UR) participants’ perceptions of institutional inclusion affected burnout, intent to continue training, and perceived stress. METHODS/STUDY POPULATION: Building Up was conducted at 25 institutions with 225 UR post-doctoral fellow or early-career faculty participants. To assess perceived institutional inclusion, participants completed a 28-item survey in the first year of follow-up. We used descriptive statistics to describe age, race/ethnicity, and gender. We used exploratory factor analysis to extract factors or domains (survey questions that grouped together). We calculated mean domain scores and used correlations to assess associations between each domain and each dependent variable (burnout, intent to continue training, and perceived stress). Demographics, correlation coefficients and associated p-values are presented. RESULTS/ANTICIPATED RESULTS: 130 of 144 eligible participants completed all questions. The mean age was 39 years (SD = 6), 83% were female, 35% identified as non-Hispanic Black, and 36% identified as Hispanic. Greater inclusivity was associated with lower burnout across 5/6 identified domains: policies (-0.3, p DISCUSSION/SIGNIFICANCE: These findings suggest that institutional inclusion is associated with differences in capacity to function among UR postdocs and early-career faculty. Inclusivity of leaders was only associated with intent to continue training. Inclusion coupled with employee support and development are important for positive outcomes.

Objectives/Goals: To validate 20 culturally tailored clinical vignettes for a translational study assessing ChatGPT’s diagnostic support in adolescent and young adult mental health. The validation ensures methodological, ethical, and cultural rigor to examine ChatGPT’s potential in collaborative clinical decision-making. Methods/Study Population: Twelve licensed clinical psychologists evaluated 50 culturally adapted vignettes using a nine-item instrument assessing diagnostic agreement (Fleiss κ), content validity (I-CVI, S-CVI/Ave), comorbidity sensitivity, difficulty, ethical sensitivity, and perceived risk. Qualitative feedback refined clarity and realism. Vignettes meeting κ ≥ 0.60, S-CVI ≥ 0.80, and ethics ≥ 3 were retained, ensuring one easy, two moderate, and one difficult case per condition. The resulting 20 validated cases will serve as standardized stimuli to assess ChatGPT’s diagnostic accuracy and its ability to collaborate effectively with clinical psychologists. Results/Anticipated Results: High inter-rater reliability (κ ≥ 0.60) and strong content validity (S-CVI ≥ 0.85) are expected. Balanced difficulty and accurate risk detection will confirm calibration. Qualitative feedback will enhance realism and cultural appropriateness. Validated vignettes will enable rigorous assessment of ChatGPT’s diagnostic accuracy and clinical decision-support potential within a translational research framework. Vignette cases will be presented to three groups: 1) ChatGPT app, 2) Clinical mental health care providers, and 3) Clinical mental health care providers collaborating with ChatGPT. Each group will provide a diagnosis and confidence level to compare accuracy, sensitivity, and collaborative value across human-AI and human-only conditions. Discussion/Significance of Impact: This validation provides a rigorous foundation for evaluating ChatGPT’s translational role in mental health diagnostics. Psychometrically and ethically sound vignettes strengthen evidence-based integration of AI tools into precision mental health care and promote responsible human-AI collaboration.

OBJECTIVES/SPECIFIC AIMS: Our primary objective is to determine the mechanism of action of myostatin on osteoblasts by measuring markers of osteoblast differentiation. With these experiments we will evaluate the effects of myostatin on an osteoblastic cell line (MC3T3 cells) and primary murine osteoblasts during baseline and hyperglycemic conditions and assess whether these effects are altered in the presence of a hyperglycemic environment. METHODS/STUDY POPULATION: Primary osteoblasts from calvaria of WT mice will be isolated and cultured per previously published protocol. MC3T3 cells (murine pre-osteoblast cell line) and primary osteoblasts will be plated in 6-well plates until they reach confluency. They will subsequently be stimulated with or without myostatin at various concentrations under control and hyperglycemic conditions. Additional experiments will assess myostatin stimulation during cell differentiation/maturation in the presence of osteogenic induction medium. Subsequently, cells will be lysed and processed for gene analysis with qPCR. Genes of interest (e.g., myostatin, RUNX2, osteocalcin etc.) will be assessed. Additionally, cells will be collected and processed for protein quantification with western blot to assess myostatin-related pathways, such as Smad2/3 and MAPK signaling. RESULTS/ANTICIPATED RESULTS: We have demonstrated that the receptor for myostatin (Activin receptor 2b, AcvR2b) is present in MC3T3 cells and we have evidence of Smad2 phosphorylation in MC3T3 cells as a result of myostatin stimulation, confirming that myostatin can exert intracellular signaling events in bone cells (Fig 1). We anticipate to observe negative effects of myostatin on differentiation of primary osteoblasts and MC3T3 cells. Specifically, we anticipate suppression of Runt-related transcription factor 2 (RUNX-2), a transcription factor known as the “ master regulator” of osteogenic gene expression and programming, as a result of signaling downstream of Smad 2/3. Additionally we anticipate downregulation of osterix and osteocalcin, two essential genes for osteoblast differentiation and activity. We anticipate that hyperglycemia will potentiate the negative effects of myostatin on osteoblastogenesis. DISCUSSION/SIGNIFICANCE OF IMPACT: We have demonstrated that myostatin can directly act on osteoblastic cells. As myostatin is a negative regulator or bone mass, its direct effects on bone cells can be detrimental to the bone health of patients with elevated myostatin levels and/or activity. There is evidence suggesting that myostatin is elevated in Type 1 diabetes, and its effects might be potentiated in a hyperglycemic environments. Future experiments will be evaluating the role of myostatin on a diabetic animal model and in humans. Our experiments provide an additional mechanism by which muscle-bone interactions could be contributing to the development of diabetic bone disease.

OBJECTIVES/GOALS: To describe an Honest Broker (HB) tool and workflow integrated with the Institutional Review Board (IRB) to automate requests, approvals and delivery of both de-identified and identified data extractions from a clinical research data warehouse (CRDW). METHODS/STUDY POPULATION: The HB tool has predefined domain tables and is closely integrated IRB for quick and easy review and approval. Investigators can access patient data using query tools, barcodes from biospecimens or build a query in TriNetX and provide the patient list as an input for the HB tool. For de-identified data extraction, the required data domain tables and date ranges can be selected and submitted in the HB tool. For identified data extractions, investigators with an approved IRB protocol can enter the protocol number and the approved date range in the HB tool. This request is automatically forwarded to the IRB for review. RESULTS/ANTICIPATED RESULTS: For de-identified data extraction, an email alert is automatically sent to the investigator once the data extract is completed. For identified data extraction, if IRB approves the request, an HB is immediately notified to release the data. Data release triggers two emails to the investigator: (1) a link to an encrypted zipped file with the requested data, and (2) a password to unlock the encrypted file. If the request is denied, the IRB sends an email to the investigator with the reason for denial and options for remediation. The entire HB workflow is accomplished in a secure environment with an audit trail from the initial data request to data download by the investigator. Since the launch of the HB tool, the time from data request to delivery is approximately an hour for deidentified data and 24 hrs for identified data. DISCUSSION/SIGNIFICANCE: The HB tool has increased successful data delivery in support of publications, grant submissions, and clinical trial recruitment. Optimization of data extraction from the CRDW through automation and integration with the IRB can minimize interaction with data analysts and IRB staff, thus accelerating the conduct of clinical research.

OBJECTIVES/GOALS: To determine if current U.S. HPV vaccination rates have decreased dysplasia prevalence enough to decrease the positive predictive value (PPV) of abnormal cervical cytology. METHODS/STUDY POPULATION: This retrospective cohort study comprised a chart review of all patients 21-35 years of age who had at least 1 Pap test result within MHealth/Fairview 2016-2018. HPV vaccination data, cervical cancer screening data and dysplasia results were abstracted. Vaccinated was defined as receiving at least 1 dose of HPV vaccine, with subgroup analyses performed for those completing vaccination per ACIP guidelines and by age of initiation dichotomized as 21+ years versus <21 years. RESULTS/ANTICIPATED RESULTS: 49,764 patients meeting study criteria were identified. Among the entire study population, 10% had abnormal cytology results during the study period. Among the 4,928 patients with abnormal cytology, PPV for CIN2+ was lower among vaccinated individuals (13% vs. 18%; p < 0.0001). Among vaccinated individuals, PPV was lower among those completing vaccination (12% vs. 16% for incomplete vaccination; p = 0.04) and among those initiating vaccination at <21 years of age (9% vs 26% for 21+y; p < 0.0001). DISCUSSION/SIGNIFICANCE OF IMPACT: Among a population with low HPV vaccine coverage, the decrease in dysplasia prevalence among vaccinated individuals is resulting in a subsequent decrease in PPV of cervical cytology, particularly in those initiating vaccination prior to 21 years of age and among those completing the series. Confirmation of these results will call for changes in screening strategies for vaccinated individuals. CONFLICT OF INTEREST DESCRIPTION: Acelity: Industry grant for an investigator-initiated industry-sponsored clinical trial. Tesaro: Site PI for industry-sponsored clinical trial. NOTE: Funding from the industries above are unrelated to the research presented in the abstract.

OBJECTIVES/GOALS: Health inequities represent complex structural and systematic processes that lead to disparate outcomes for populations or subgroups within populations. This project aims to improve the available structural and systematic approaches to the study of such inequities at the population level. METHODS/STUDY POPULATION: Using examples from diabetes research, two critical factors that may impact the validity or utility of health equity models will be examined; and proposed methodological approaches to offsetting potentially resulting biases will be offered. The factors include: (1) inequitably missing and misclassified data in large datasets and (2) the presumed positioning of socially constructed variables such as race, ethnicity, and gender within modeled structural and systematic mechanisms. This examination intersects theories and praxis in epidemiological modeling and health equity promotion with the goal of advancing rigorous, equity-focused epidemiological methods. RESULTS/ANTICIPATED RESULTS: Inequitably missing and misclassified data are generally expected to obscure inequities. Treatment of missing or misclassified data as informative measures of inequity is expected to partially offset this bias. The implicitly modeled components of socially constructed variables are expected to be non-uniform across structural and systematic mechanisms of inequity. Models that apply these variables as informatively heterogeneous constructs, using multi-phase analyses to test modeling assumptions and to assess intersectionality, may provide better context about the mechanisms by which inequity has been distributed and, perhaps, by which equity may be achieved. DISCUSSION/SIGNIFICANCE: Equitable epidemiological methods are essential to the advancement of evidence-based health equity on the population level. Potential structural or systematic inequities in large-scale datasets and traditional data analyses should be thoughtfully reviewed through a health equity lens.

OBJECTIVES/SPECIFIC AIMS: This study is part of a parent grant evaluating antidiabetic medications and risk for heart failure in an observational cohort of Veterans with type 2 diabetes (T2DM). Confounding by indication remains a concern in many observational studies of medications because difficult to measure confounders such as frailty may influence prescribing of different medications based on patient characteristics. Frailty is a multidimensional syndrome of loss of reserves (energy, physical ability, cognition, health) that gives rise to vulnerability to adverse outcomes. The objective of this study is to determine if frailty is a potential confounder in Veterans with T2DM, that is, independently associated with exposure to a specific antidiabetic medications and hospitalization for decompensated heart failure. METHODS/STUDY POPULATION: We conducted a cross-sectional study of patients with diabetes who were hospitalized within the Veterans Health Administration (VHA) Tennessee Valley Healthcare System from 2002 to 2012. Inclusion criteria were: age 18 years or older, receive regular VHA care (prescription fill or visit at least once every 180 d), a diagnosis of T2DM. A probability sample of HF and non-HF hospitalizations was collected. HF hospitalizations were selected on the basis of meeting either a primary diagnosis code (ICD-9) and/or disease related group (DRG) code for HF. For each hospitalization using a standardized chart abstraction tool, data was abstracted on: antidiabetic medication(s), patient frailty status, and reason for hospitalization (HF or non-HF). Antidiabetic medication regimens were categorized as follows: no medication treatment, metformin alone, sulfonylurea alone, insulin alone, insulin and one oral agent, and all other regimens. Patient frailty status was measured using a modified version of the Canadian Health and Aging frailty index (FI), which generates a score (range 0–1) by dividing the number of deficits present by the number of deficits measured. Established categories for FI scores are: non frail ≤0.10, vulnerable 0.10–0.21, frail 0.22–0.45, and very frail >0.45. Patient frailty status at the time of hospitalization was used as a surrogate for patient frailty at the time of prescription of antidiabetic medication; this is a limitation of this approach. Hospitalizations were classified as HF hospitalizations if 2 major or 1 major and 2 minor Framingham criteria were present. FI was compared across antidiabetic medication regimen categories and hospitalization type using analysis of variance (ANOVA) and Student t-test, respectively. RESULTS/ANTICIPATED RESULTS: Of the 500 hospitalizations reviewed, 430 patients had confirmed diabetes diagnosis, adequate data to calculate FI scores, and were included in this analysis. Patients were on average 66.9 (10.9) years old; 99% male and 75% were White. Overall, 268 patients (62.3%) were categorized as frail or very frail. The mean FI score was 0.23 (SD 0.07). FI scores were highest in patients receiving insulin alone (mean 0.26) compared with patients receiving metformin alone (mean 0.22), sulfonylurea alone (mean 0.23), or no medication (mean 0.22). The lowest mean frailty score was seen in patients taking all other drug combinations, 0.19. The differences across these patient groups were statistically significant with p<0.01. Further, 75% of patients on insulin alone were frail or very frail compared with 68% on sulfonylurea alone, 58% on metformin alone, and 58% on no medication. Framingham criteria for acute HF were present for 318 of 430 patients (74.0%). FI scores were higher in patients hospitalized for HF compared with non-HF hospitalizations (mean 0.24 vs. 0.21, p<0.01). A higher proportion of patients hospitalized for HF were classified as frail or very frail compared with those hospitalized for non-HF diagnosis (66.4% vs. 50.9%, p<0.01). DISCUSSION/SIGNIFICANCE OF IMPACT: This study demonstrates that certain antidiabetic medications are associated with patient frailty. In addition, those patients admitted for HF have higher FI scores than those admitted for non-HF diagnoses. Further investigation is planned to assess the degree to which frailty is captured by traditional covariates used in observational studies.

Objectives/Goals: 1) Determine the association between wildfire smoke-driven PM2.5 and risk of persistent respiratory symptoms and repeated asthma exacerbations after the acute wildfire period among adults with asthma. 2) Examine how measures to reduce personal exposure to wildfire smoke, including avoiding outdoor activities, modify this association. Methods/Study Population: This is a retrospective study of adults with asthma in WHAT-NOW, a cohort study of people living in Northern California during the 2018 Camp Fire. Daily smoke-driven PM2.5 was estimated for each participant based on their home address or evacuation location. We examined the association between mean PM2.5 exposure and the presence of respiratory symptoms at both the time of the survey (6–16 months post-wildfire) and at least one other post-wildfire time-period, as well as whether they had a medically attended respiratory illness (saw a doctor, visited the ER, or were hospitalized for a respiratory symptom). We examined the interaction of PM2.5 with spending time outdoors during the wildfires. Poisson regression models with robust standard errors were adjusted for age, sex, race, smoking, allergies, and education. Results/Anticipated Results: Among 337 adults with asthma in the WHAT-NOW cohort, one standard deviation higher smoke-driven PM2.5 was associated with higher risk of any persistent respiratory symptom (risk ratio (RR) 1.38, 95% CI 1.07 – 1.78) and having at least one medically attended respiratory illness (RR 1.33, 95% CI 1.07 – 1.65), but not significantly associated with repeated asthma exacerbations (RR 1.30, 95% CI 0.92 – 1.81). However, there was a significant interaction between PM2.5 and outdoor activities during the wildfire on the outcomes of any persistent respiratory symptoms (p = 0.041) and repeated asthma exacerbations (p = 0.028). The association between PM2.5 and repeated asthma exacerbations was greater among people who spent time outdoors (RR 3.36, 95% CI 1.47 – 10.23) than those who did not (RR 1.00, p = 0.99). Discussion/Significance of Impact: This study provides evidence that exposure to wildfire smoke increases respiratory morbidity among adults with asthma beyond the acute wildfire period. Additionally, it suggests that avoiding outdoor activities on smoky days can significantly decrease the risk of future repeated asthma exacerbations associated with smoke exposure.

OBJECTIVES/SPECIFIC AIMS: The objective of the present study is to determine if decreased adiponectin and increased leptin levels are associated with the development of MetS and identifiable endothelial dysfunction in a cohort of Hispanic pre-pubertal children. To do so we propose the following aims: (1) To measure expression of adiponectin and leptin levels in a Hispanic pre-pubertal cohort and determine their correlation with features of the MetS. (2) To perform proteomic analysis in a Hispanic pre-pubertal cohort. (3) Evaluate early onset of endothelial dysfunction and its correlation with expression of adiponectin and leptin levels in a Hispanic pre-pubertal cohort. METHODS/STUDY POPULATION: A cross-sectional pilot study will obtain a random representative sampling of children aged 6–12 years from all geographical areas of Puerto Rico. Children will be assessed regarding pre-pubertal status through Tanner staging and later divided into pre-MetS Versus MetS groups as well as controls. MetS will include children meeting 3 or more of the current International Diabetes Federation (IDF) criteria. Pre-MetS will include children with at least 1 criterion for MetS. Anthropometric data, blood pressure readings, ultrasound-based noninvasive testing for endothelial dysfunction, and laboratory assays will be performed to the study population and data analyzed for correlation. Total adiponectin and leptin levels will be measured using a commercially available quantitative sandwich enzyme-linked immunoassay test. The study will be submitted to the University of Puerto Rico Medical Sciences Campus’ Institutional Review Board (IRB) for approval. Written consent and assent will be obtained from parents and children respectively to ensure patient anonymity. RESULTS/ANTICIPATED RESULTS: We hypothesize that low levels of adiponectin and high levels of leptin will correlate with features of the MetS as defined by the IDF consensus statement, as well as with clinical features of MetS in undiagnosed Hispanic pre-pubertal youth. We also hypothesize that non-invasive testing of endothelial function will correlate both with clinical features of the MetS and with low levels of adinopectin and high levels of leptin. DISCUSSION/SIGNIFICANCE OF IMPACT: The correlation of findings suggestive of endothelial dysfunction and biomarker expression (mainly adiponectin and leptin levels) in a pre-pubertal cohort has yet to be established and could also provide information regarding early atherogenesis in otherwise unidentified youth at risk. Therefore, by using a proteomic approach, this study aims to measure associations between clinical features of the MetS and expression of proteins associated with an adverse cardiometabolic profile in a Hispanic pre-pubertal population. We will concurrently measure the degree of endothelial dysfunction and evaluate whether a correlation exists between previously mentioned protein expression and early onset of dysfunction.