Recent reports on the burden of cardiovascular disease (CVD) in the USA indicate that despite significant declines in CVD mortality in the late 20th century, this decline is now decelerating and may be worsened by inequalities in health care. Social factors contribute to most of the cardiovascular health disparities documented to date. Hispanics/Latinos and African-Americans share a higher prevalence of cardiovascular risk factors and experience higher rates of poverty and social stressors than non-Hispanic Whites. We propose that the use of social and behavioral data beyond basic and sometimes loose identifiers of race/ethnicity, educational attainment, and occupation would inform clinical practice and greatly facilitate the provision of adequate guidance and support to patients regarding continuity of care, adherence to medications and treatment plans, and engagement of participants in future research. This perspective briefly highlights factors deemed to be critical for the advancement of Hispanic/Latino health and delineates pathways toward future applications.

OBJECTIVES/GOALS: The New Jersey Alliance for Clinical & Translational Science conducted an observational study of current clinical research training to establish a comprehensive infrastructure. The goal was to develop a cross training catalogue of available resources for trainees to fill gaps in their Clinical Research training. METHODS/STUDY POPULATION: The focus of this observational study is the evaluation of (1) available clinical research training programs, (2) training areas aligned with the Joint Task Force competency domains, (3) specific target audiences who utilize trainings offered nationally, and (4) availability of resource toolboxes. This assessment was extracted by clicking on each of the hubs Education & Training and Resources tabs. The determination of the training areas and competency skills that aligned with the JTFs eight domains was confirmed by reviewing the course description on each website. RESULTS/ANTICIPATED RESULTS: CTSA hub website analysis of each hub revealed that each CTSA hub region has their own regional focus on training areas targeted for specific audiences. The validation of the NJ ACTS Education and Offering Inventory study allowed authors to categorize available training programs which significantly showed that Basic Training Programs targeted 22% of Undergraduate Students, Community Engagement Training Programs targeted 15% of Early Career Faculty, Experiential Training targeted 20% of Early Career Faculty and Undergraduate Students, Scholarly & Certificate Training targeted 20.5% of Graduate Students, and Graduate/Post Graduate Programs targeted 22.5% of Graduate Students and KL2 scholars. DISCUSSION/SIGNIFICANCE: This information helps build the infrastructure of a National Cross-Training Resource Platform in aid to encourage trainees to take advantage of available trainings across the CTSA Consortium.

OBJECTIVES/GOALS: Human placentation requires complex coordination between maternal and fetal cell types but remains incompletely understood. We hypothesize that uterine natural killer (uNK) cells, an immune cell type that increases in abundance during the implantation window, is essential for appropriate implantation and placentation. METHODS/STUDY POPULATION: We plan to examine stromal cell (SC) decidualization, spiral artery remodeling, and EVT invasion, processes vital for early pregnancy establishment, in the presence or absence of secretory phase uNK cells. Fetal extravillous trophoblasts (EVTs) will be isolated from first trimester pregnancy tissue; maternal SCs, endothelial cells (ECs) and uNK cells will be obtained from secretory phase uterine tissue. SCs will be placed in monoculture and coculture with uNK cells and prolactin will be measured to evaluate decidualization. To study EVT invasion, we will utilize our novel “implantation-on-a-chip” device to determine how addition of uNK cells affects EVT migration through a collagen-matrigel matrix. In this system, we will also examine spiral artery remodeling with or without uNK cells via TUNEL staining. RESULTS/ANTICIPATED RESULTS: We anticipate that uNK cell addition to SCs will lead to a significant increase in SC prolactin levels, suggesting a role of uNK cells in endometrial decidualization. In vitro, we expect the addition of uNK cells will increase EC apoptosis and promote EVT invasion. DISCUSSION/SIGNIFICANCE OF IMPACT: Although decidual NK cells are known to participate in placentation, the role of pre-pregnancy uNK cells is unknown. uNK cell involvement in processes important for the earliest stages of pregnancy would provide a potential marker for abnormal placentation and offer avenues for intervention to decrease placentation associated perinatal morbidity.

OBJECTIVES/SPECIFIC AIMS: Thoracic endovascular aortic repair (TEVAR) is more effective in remodeling the dissected aorta in acute versus chronic type B aortic dissection (TBAD). It has been hypothesized that this is due to differences in dissection flap biomechanical and structural properties but has not been confirmed in explanted human aortic tissue. We aimed to characterize and compare differences in tissue biomechanics and microstructure between acute and chronic dissection flaps that may underlie these findings. METHODS/STUDY POPULATION: Dissection flaps were obtained at time of operative repair for patients presenting for open aortic replacement to treat acute type A (ACUTE, n=7) or chronic type B (CHRONIC, n=7) aortic dissection. Given that the current treatment modality for acute complicated TBAD is TEVAR, it was not feasible to acquire acute TBAD flaps for analysis. Tissues were cryopreserved and subjected to biaxial tensile testing in the circumferential and longitudinal directions. Stiffness was quantified by the tangent modulus (TM) in the low and high linear regions of the compiled equibiaxial response curves for each cohort. Extensibility was defined as the intersection of the fitted line from the high linear region with the x-axis, and the degree of anisotropy (DA) was defined as the mean absolute percentage error of the strains in both directions. Flap architecture and collagen fiber organization were also compared between groups using two-photon microscopy. RESULTS/ANTICIPATED RESULTS: Average age of dissection flaps were 3.4±3.4 days in ACUTE and 1,868.7±1,354.0 days in CHRONIC (p=0.011). There were no differences in age, co-morbidities, maximum aortic diameter, and aortic wall thickness. ACUTE exhibited an anisotropic stress-strain response with increased extensibility longitudinally than circumferentially (0.18 vs. 0.09, p=0.022, DA=0.67) while CHRONIC demonstrated an isotropic response with similar extensibility in either direction (0.11 vs. 0.12, p=0.606, DA=0.26). CHRONIC and ACUTE had comparable stiffness in the circumferential direction (TMlow 439.92 vs. 541.08, p=0.729, and TMhigh 1585.19 kPa vs. 1869.35 kPa, p=0.817). In the longitudinal direction, CHRONIC was significantly stiffer than ACUTE (TMhigh 8347.61 kPa vs. 1201.34 kPa, p=0.049) (FIGURE). Microscopy corroborated these findings with greater collagen fiber organization circumferentially than longitudinally in ACUTE and increasing fibrosis, collagen predominance, and straightening of collagen fibers in CHRONIC. DISCUSSION/SIGNIFICANCE OF IMPACT: Compared to ACUTE, CHRONIC exhibited loss of anisotropy with increased tissue stiffness in the longitudinal direction. Increased dissection flap fibrosis and decreased compliance may explain the worse outcomes for aortic remodeling after TEVAR in chronic TBAD. This study offers biomechanical support for early TEVAR in the acute phase of uncomplicated TBAD.

OBJECTIVES/GOALS: Donor hearts are transported in cold storage (CS) and undergo ischemia-reperfusion injury (IRI) when transplanted. IRI injures microvascular endothelial cells (EC), heightens the immune response, and has been associated with increased autophagy. We aim to understand the changes in autophagy during CS and IRI and its impact on EC immunogenicity. METHODS/STUDY POPULATION: To study autophagy changes during IRI, immunoblotting for autophagy markers was performed in mouse cardiac ECs (MCECs) lysates. MCECs were in a cold preservation solution in a hypoxic chamber for 6 hours(h) and warm conditions with culture medium for 24 h. MCECs, under standard conditions, served as controls. Secreted interferon-gamma (IFN-γ) levels were quantified via ELISA to study autophagy and EC immunogenicity. MCEC-sensitized CD8+ T-cells were isolated from C57BL/6 spleens and co-cultured with MCECs pre-treated for 16 h with rapamycin or starvation, autophagy inducers, or chloroquine, an autophagy inhibitor under normal or IRI conditions. MCECs without any treatment served as controls. RESULTS/ANTICIPATED RESULTS: To determine autophagy levels in IRI, immunoblotting of MCEC lysates revealed a significant increase (P<0.01) in the established autophagy marker, LC3, at early time points post-reperfusion compared to NT conditions, indicating more autophagosome formation during CS and IRI. To assess the role of autophagy in EC immunogenicity, the co-culture experiment revealed that autophagy induction in MCECs under NT and HCS conditions with rapamycin had a 74.9-fold and 51.5-fold reduction of IFN-γ (pg/mL), resepectively, compared to the non-treated controls. In contrast, autophagy inhibition in MCECs with chloroquine resulted in 1.82-fold increase of IFN-γ compared to untreated controls. This suggests a protective role of autophagy in ECs during IRI. DISCUSSION/SIGNIFICANCE: We observed that autophagy may be protective during IRI by mitigating EC immunogenicity. Thus, pharmacologically modulating microvascular EC autophagy in donor hearts prior to transplantation may mitigate insults incurred during CS and IRI.

Objectives/Goals: Children with intestinal failure rely on central venous catheters (CVCs) for at-home delivery of hydration and nutrition to sustain life. CVC associated infections are the leading driver of morbidity in this population. It is currently unknown what challenges caregivers of this patients face for at-home infection prevention tasks. Methods/Study Population: This is a qualitative analysis of the clinical implementation of at-home CVC care. Participants will be caregivers of children with intestinal failure in the Children’s Hospital of Los Angeles’ Intestinal Rehabilitation Program (a program with a large Spanish speaking population). The study team will conduct, record, and transcribe four synchronous remote focus groups using Zoom, two in English and two in Spanish, with 4–6 participants per focus group (n = 16–24). Questions will focus on caregiver experiences of CVC care. Transcripts will be analyzed using rapid qualitative analysis, an implementation science-oriented approach. The study team will review transcripts and summarize key points into matrices to examine relevant themes and quotes efficiently and systematically, using midpoint data analysis. Results/Anticipated Results: Illustrative quotes of all themes and domains discussed by caregivers will provide a person-centered overview of the specific types of facilitators and barriers to infection prevention task performance. Focus groups will be conducted in January 2025 with analysis in February. Facilitators and barriers will be mapped onto the Capability, Opportunity and Motivation behavioral model (COM-B), allowing for additional topics participants introduce by participants. Facilitators and barriers will affect all domains of the COM-B model and may also describe factors that cross multiple domains or fall outside the COM-B domains. Facilitators and barriers may differ between English and Spanish speaking caregivers. Discussion/Significance of Impact: Identifying facilitators of and barriers to infection prevention tasks will allow clinicians to maximize infection prevention efforts and improve quality of life for caregivers of children with intestinal failure. Additionally, this bilingual study will have improved external validity and address potential language-related barriers to care.

ABSTRACT IMPACT: This work identifies host immune perturbations in sepsis mortality that suggests targets for a precision medicine paradigm in the host response to infection. OBJECTIVES/GOALS: To compare the early whole blood transcriptome during sepsis between 30-day survivors and non-survivors in the Intensive Care Unit (ICU), and to evaluate for pathway enrichment that might explain sepsis lethality. METHODS/STUDY POPULATION: We enrolled 162 sepsis patients in the first 24 hours of ICU admission, particularly targeting individuals requiring vasopressors. Peripheral whole blood was collected in PAXgene vacutainers. Isolated RNA was analyzed with Affymetrix Human Genome ST 2.0 microarray. Differential gene expression was performed with Bioconductor/R/limma using log2 fold-change +/-0.6 as a threshold for differential expression, and a Benjamini-Hochberg adjusted p-value <0.05 to declare significance. Functional gene enrichment was performed using the Gene Ontology (GO) database with PANTHER overrepresentation test (Fisher’s Exact) on all transcripts with adjusted p-value <0.05. Pathways analysis was performed with the Reactome Project using the raw fold change and significance data to identify dysregulated pathways. RESULTS/ANTICIPATED RESULTS: There were 58 non-survivors (36% mortality). We identified 39 genes as differentially expressed between sepsis non-survivors and survivors; 31 were upregulated in non-survivors and 8 had reduced expression. Several of the most overexpressed transcripts are neutrophil-specific, including LCN, MPO, OLF4M4, DEFA3, and DEFA4. A functional gene overrepresentation test further supports this finding, as the most enriched gene ontologies were neutrophil-mediated killing, neutrophil cytotoxicity, neutrophil extravasation, and respiratory burst, all demonstrating higher than 10-fold enrichment and FDR < 0.02. Pathway analysis of the peripheral blood transcriptome was notable for immune response derangement, specifically downregulation of both innate and adaptive immune pathways (FDR < 0.00001). DISCUSSION/SIGNIFICANCE OF FINDINGS: We identified increased expression of neutrophil-related genes in sepsis non-survivors, replicating candidates previously identified in pediatric sepsis mortality and ARDS. These immune perturbations in sepsis mortality may represent key targets for eventually employing precision medicine strategies in sepsis.

OBJECTIVES/GOALS: Anti-cancer therapies, such as chemotherapy, can induce senescence. Senescent cells may produce factors that can promote tumor progression. In this study, we will investigate the effect of senolytics and anti-cancer treatment on fibroblasts, which are a part of the tumor microenvironment, and patient-derived colorectal cancer organoids. METHODS/STUDY POPULATION: We will induce senescence in fibroblast lines via irradation. Induction of senescence will be confirmed by monitoring SASP production, changes in morphology and proliferation rates, and senescence-associated b-galactosidase activity. To investigate the efficacy of senolytics on senescence-induced fibroblasts and CRC tumor organoids, we will creat a dose response curve and calculate IC50 values for proliferating fibroblast, senescent fibroblasts and CRC organoids. To identify the synergistic effects of anti-cancer and senolytic compounds, including Navitoclax and Dasatinib, on fibroblasts and CRC organoids, we will create dose matrixes using senolytics at concentrations that were shown to have senolytic activity and drugs from an anti-cancer library. RESULTS/ANTICIPATED RESULTS: If senescence is induced in the fibroblast lines, we expect to see no changes in confluency over 4 days, the morphology will change from a thin, spindly shape to a flattened shape, and senescence-associated b-galactosidase activity will be observed. After the fibroblast lines are treated with potential senolytic compounds, we would expect to see decreased viability in the senescence-induced fibroblast lines when compared to proliferating fibroblast lines. We predict that the viability of CRC organoid lines will slightly decrease at high concentrations of the senolytic due to overall toxicity. We expect that the senolytic and anti-cancer compounds will have a synergistic effect. Senolytic activity could reduce the senescent cell population that was developed in response to anti-cancer therapy. DISCUSSION/SIGNIFICANCE: There is an increased interest in identifying compounds that selectively promote apoptosis in senescent cells. This study uses a cell-based approach to validate senolytic activity of compounds with senolytic potential in senescence-induced fibroblast lines and investigates the synergistic effects of senolytics and anti-cancer compounds on CRC.

Objectives/Goals: 1) Discuss process of pilot integration of Community Health Worker (CHW) services as a component of patient-centered healthcare service delivery in 3 clinic models. 2) Summarize profiles of patients who self-select to utilize CHW services. 3) Discuss social determinants of health impacts of underserved and historically marginalized populations. Methods/Study Population: The priority population consists of individuals living in Mobile AL at or below poverty level. USA Health Center for Healthy Communities (CHC) piloted the integration of CHW services at USA Health Stanton Road Clinic (SRC), at USA Student Run Free Clinic (SRFC), and as part of a Medi Hub Outreach Clinic with the historically underserved MOWA Choctaw native American population. SRC is a high-utilization clinic for uninsured or underinsured patients across the breadth of the Gulf Coast. The other 2 sites serve similar clientele. Social determinants of health (SDOH) screenings at intake facilitate CHW referral for a clients’ unique needs for support at healthcare or social care agencies. Referral summaries can then be used to guide planning, community collaborating partner intervention, and clinical quality certification, Results/Anticipated Results: Results include identification of referrals process by which CHWs are able to provide culturally competent support to persons accessing healthcare services at the 3 clinic models identified. Identification of top SDOH needs that preclude access to care among the patients served during a 24-month pilot period, e.g., (i) housing insecurity, (ii) food insecurity, (iii) transportation, (iv) health Ins, and (v) pharmacy access and payment assistance. Discussion of beneficial impacts for health care service delivery with other members of the multidisciplinary clinical teams as recorded referrals can be used to guide planning, clinic certification efforts Discussion/Significance of Impact: Patient utilization of CHW services though self-selective offers opportunities for equity in access to care services from direct SDOH impacts, where CHWs act as responsive resource coordinators within the multi-disciplinary service delivery team.

OBJECTIVES/GOALS: Our lab previously identified a population of vascular smooth muscle (SMC)-derived progenitor cells (AdvSca1-SM) which expand robustly in response to disease and can differentiate into multiple cell types. We now aim to define the role of these AdvSca1-SM cells in atherosclerotic plaque progression. METHODS/STUDY POPULATION: Goal one uses SMC lineage tracing mice and a model of atherosclerosis to track reprogramming of SMCs to AdvSca1-SM cells in the setting of disease. Arteries are analyzed using flow cytometry and immunofluorescence to quantify changes in number of mature SMCs and AdvSca1-SM cells. Goal two uses AdvSca1-SM lineage tracing mice with high cholesterol-induced atherosclerosis and plaque neovascularization. Arteries are analyzed to quantify expansion of AdvSca1-SM cells, subsequent re-differentiation into mature SMC, endothelial cells, or macrophages, and contribution to plaque neovascularization. Mechanistic findings from both goals are being investigated in diseased human coronary arteries. RESULTS/ANTICIPATED RESULTS: Flow cytometry from SMC lineage tracing mice revealed a 7- to 13-fold expansion of AdvSca1-SM cells in carotid arteries (p<0.001) and aortas (p = 0.03) after 6 weeks of western diet; no differences in macrophage numbers were observed. Additional SMC and AdvSca1-SM cell lineage tracing mice are on atherogenic diets to assess early and advanced atherosclerosis. We predict that AdvSca1-SM cells will contribute to macrophage accumulation as well as plaque neovascularization in the setting of severe atherosclerosis. Translational relevance of mechanisms driving SMC reprogramming and AdvSca1-SM cell contribution to plaque progression are being applied to studies of diseased human coronary arteries. DISCUSSION/SIGNIFICANCE OF IMPACT: Our data suggest a role for AdvSca1-SM cells in atherosclerosis. Ongoing work will clarify the mechanisms driving plaque-associated AdvSca1-SM expansion and define the ultimate fates of these cells. In vivo modulation of this process could provide the basis for future anti-atherosclerotic therapies. CONFLICT OF INTEREST DESCRIPTION: AD - CCTSI TOTTS TL1TR002533; SL - 18POST34030397 from the American Heart Association; AJ – no conflicts; KS - 1F31HL147393 from the National Heart, Lung, and Blood Institute, NIH; MM – no conflicts; RT – no conflicts; KSM – no conflicts; RAN - R01CA236222 from the National Cancer Institute, NIH, and 2018-03 from the Lungevity Foundation; and MCMW-E - R01 HL121877 from the National Heart, Lung, and Blood Institute, NIH, and 25A8679 from the Chernowitz Foundation.

OBJECTIVES/GOALS: Use an easily accessible medium to educate life science researchers and academic innovators interested in the commercialization of academic research at the University of Michigan (UM). METHODS/STUDY POPULATION: Life science research investigators and academic innovators interested in research commercialization and technology development from across the state of Michigan were invited to attend the Idea to Impact: The Translation & Commercialization of Academic Research webinar series, presented by Fast Forward Medical Innovation at the University of Michigan. The webinar series outlined the significance and critical milestones of developing novel therapeutics, medical devices, diagnostics, and digital health innovations, as well as essential collaborations with industry partners to translate a research-based idea into a product of impact. RESULTS/ANTICIPATED RESULTS: 113 investigators and innovators from 28 different institutions, organizations, and companies, registered for the webinar series. Results (N=24) of an evaluation immediately following each webinar revealed that 100% of respondents strongly agreed or agreed that the series was effective in helping them to identify and describe commercialization resources, including funding, education, and mentorship, available at the University of Michigan and within the state. Participants stated that they “loved the practical information” “shared” and that the series was a “great overview that inspired a lot more questions.” The Fast Forward Medical Innovation team was then able to consult with participants to connect them with additional resources. DISCUSSION/SIGNIFICANCE: The data suggests that easily accessible and digestible commercialization education can make navigating the academic entrepreneurial ecosystem easier for investigators and innovators. The recorded webinar series, Idea to Impact: The Translation & Commercialization of Academic Research, serves this purpose.

OBJECTIVES/GOALS: Adult spinal deformity is commonly treated by spine surgeons. Patient treatment planning and surgical candidacy are dependent on static measurements and inconsistent heuristics which lead to high complication rates and poor outcomes. This study tests the role of supplemental longitudinal and dynamic patient data in improving surgical planning. METHODS/STUDY POPULATION: Ten adult spinal deformity surgeons at Johns Hopkins Hospital were interviewed for 30 minutes by the study team. The script was reviewed by the institutional review board to alleviate any risk of bias. Two patient sets were curated utilizing previously treated, anonymized patient data sets from a non-surveyed practitioner. Each patient set was coupled with relevant radiographic imaging (MRIs, CTs, and plain radiographs) and pertinent clinical information that is collected in a standard clinic visit. Surgeons were presented with a patient and asked to note their specific surgical plan. Subsequently, surgeons were presented with four sets of supplemental dynamic spine data and asked to note their surgical plan for each set. Shaprio-Wilks and Mann-Whitney U tests were used to assess data normality and nonnormality. RESULTS/ANTICIPATED RESULTS: Preliminary data has shown inconsistency in both surgical selection and surgical type amongst physicians when presented with initial clinical findings and radiographic reports for base patient cases. There was minimal consensus among surgeons on the number of levels fused and interbody spacer usage. Early results show that dynamic spine data may be beneficial in creating consistency between surgeons, despite inter-surgeon variability in surgical planning without this data. Posture, pain location, pain severity, and quantified activity throughout the day have been referenced as the most useful dynamic spine data to consider. Amongst all providers, the availability of dynamic spine data resulted in a change in surgical planning. DISCUSSION/SIGNIFICANCE: Recent publications have shown that spine surgery patient candidacy and surgical planning are dependent on heuristics. This has led to inconsistencies amongst surgeon preferences and increases in improper patient selection for procedures. Incorporating longitudinal dynamic data may lead to increased consistency and improved patient outcomes.

OBJECTIVES/GOALS: Multiple sclerosis (MS) is a chronic inflammatory disease of the central nervous system. MS affects more than two million people worldwide, resulting in physical disability, cognitive impairment, and decreased quality of life. We are investigating the role of senescent cells, a hallmark of aging, in inflammation and disease progression in MS. METHODS/STUDY POPULATION: Female mice on a C57BL/6 background were subjected to the Hooke Laboratory EK-2110 experimental autoimmune encephalomyelitis protocol (EAE; n=10) to induce hind limb paralysis, and matching control mice received no injections (naīve; n=10). Immunofluorescent staining was used to visualize senescence cells and their localization in spinal cord tissue sections from naīve and EAE mice based on antibody detection of cell surface or intracellular proteins. Tissue sections from each group were analyzed in duplicates for each antibody (n=3-4/group). Flow cytometry was performed to immunophenotype the senescence cells using conjugated fluorescent antibodies to cell surface or intracellular proteins (n=5/group). RESULTS/ANTICIPATED RESULTS: Immunostaining demonstrated an increase in the cell senescence markers p16 (15-fold; unpaired t-test, p=0.01; n=3) and p21 (15-fold; unpaired t-test, p=0.003; n=3) in the spinal cord of EAE mice compared to naīve mice. Next, we showed that p16+ and p21+ cells increase with disease progression in the meninges adjacent to the ventral demyelinating lesions (one-way ANOVA, p=0.0009; n=3/group). We further found that 30±9.7% of M2 macrophages, a subset of myeloid cells, express p21 in the spinal cord of EAE mice by using flow cytometry analysis compared to only 5±1.6% in naīve mice (unpaired t-test, p=0.002; n=5/group). DISCUSSION/SIGNIFICANCE: Our findings demonstrate that senescent cells accumulate in the meningeal compartment following EAE induction, suggesting that decreasing senescent cell burden is a promising avenue in delaying disease progression and preventing neuroinflammation in MS.

OBJECTIVES/SPECIFIC AIMS: Diabetes mellitus (DM) is a risk factor for the development of microvascular complications. We sought to determine the association between diabetes mellitus status and microvascular circulatory disease, as measured in the sublingual capillary bed. METHODS/STUDY POPULATION: We prospectively recruited adults with cardiovascular (CV) risk factors or established CV disease, with and without DM, who were referred for invasive coronary angiography at an urban tertiary care medical center. All participants underwent non-invasive sublingual sidestream darkfield microscopy. The primary outcome was the perfused boundary region (PBR), a measure reflecting the extent to which red blood cells (RBC) penetrate the sublingual glycocalyx in vessels between 5 and 25 µm in width. RESULTS/ANTICIPATED RESULTS: 57 participants were enrolled. The mean age was 66.1 ± 11.1 years and a majority of participants (66%) were men. DM was present in 18 (31.6%) participants. Sublingual PBR measurements were not different between participants with and without DM overall (1.93µm vs. 1.96µm, p=0.63) or in vessels with high flow (1.48µm vs. 1.59µm, p=0.08). No differences in capillary RBC filling (72.9% vs 73.0%, p=0.95) or perfused microvascular density (3112 vs. 3236 µm/mm2, p=0.32) by DM status were observed. DISCUSSION/SIGNIFICANCE OF IMPACT: In a population of adults with CV risk factors or disease, DM was not associated with impaired sublingual microvascular glycocalyx. Additional investigation into diabetes-induced microvascular impairment is warranted.

OBJECTIVES/GOALS: Participants will learn about the Connections IN Health partnership. Participants will learn about the mixed-methods, multi-level evaluation model used to evaluate health coalitions. Participants will learn about how the Connections IN Health partnership promoted coalition growth among Indiana chronic disease coalitions. METHODS/STUDY POPULATION: Chronic disease coalitions under the Connections IN Health (CINH) partnership were evaluated using a novel mixed-methods, multi-level evaluation framework that triangulates coalition functioning and effectiveness (F/E) surveys, social network analysis (SNA), and health data. F/E and SNA surveys were distributed to active coalition members of the chronic disease coalitions before, 1 year, and 2 years after CINH was implemented. F/E data was analyzed to detect significant changes in coalition members’ perceptions of coalition functioning and effectiveness using unpaired, two sample t-tests in R. SNA data was analyzed to detect changes in partnership networks over time, such as membership growth and strength of connections using R software. County-level health data will be evaluated at year 5. RESULTS/ANTICIPATED RESULTS: The results of our longitudinal analysis of perceptions of functioning and effectiveness demonstrated significant increases in perceived leadership, satisfaction with the coalition, outcome efficacy, and members’ commitment to the coalition after Connections IN Health (CINH) was implemented. Social network analysis (SNA) demonstrated growth of coalition membership and strength of connections over time, including health information sharing, formal relationships, trust, and communication. SNA results also demonstrated increases in centralization and clustering among local vs. statewide organizations in coalition networks over time. Results were shared back with coalition members to make data-driven decisions about future coalition processes and functioning. DISCUSSION/SIGNIFICANCE: Health coalitions are a nationally recognized vehicle to promote and translate evidence-based practices to communities; however, evidence to support their effectiveness is limited at best. Our results provide evidence of CINH’s efficacy and identifies factors required to build effective and sustainable health coalitions.