OBJECTIVES/GOALS: The overall goal of this study was to determine the effect of early life stress (ELS) on the intestinal CD4+ T cell immune compartment, at homeostasis and after induction of experimental Inflammatory Bowel Disease (IBD). METHODS/STUDY POPULATION: We used a mouse model of ELS, maternal separation with early weaning (MSEW). We used IL-10 reporter mice to enable analysis of IL-10-producing cells. Mice were examined on postnatal day 28 to determine the impact of ELS on gut regulatory T cells. Plasma levels of corticosterone (rodent stress response hormone) was determined by ELISA. Colitis was induced in MSEW and normal rear (NR) mice via intraperitoneal injection of α-IL-10R every 5 days until day 15. Mice were euthanized on days 20 and 30. Colonic tissue sections were stained for histological analysis. Remaining tissue was further processed for flow cytometric analysis of CD4+ T cells and innate lymphoid cells. RESULTS/ANTICIPATED RESULTS: Plasma corticosterone was elevated in MSEW mice compared to their NR counterparts at 4 weeks of age. We observed that the MSEW stress protocol does not affect the baseline colonic CD4+ T cell or innate lymphoid cell populations. There was a reduction in the intestinal CD4+ T cells and regulatory T cells on day 20 in α-IL-10R MSEW mice compared to NR counterparts. This difference disappeared by day 30. Histological scoring showed no difference in disease severity between α-IL-10R treated MSEW and NR mice on day 20. However, on day 30, when α-IL-10R NR mice are recovering from colitis, MSEW mice showed persistent histological inflammation, mainly attributable to sustained epithelial damage. DISCUSSION/SIGNIFICANCE OF IMPACT: Our results suggest that ELS prolongs intestinal inflammation and impairs epithelial repair. Future studies will focus on elucidating the mechanisms responsible for ELS-dependent impairment of mucosal repair in experimental colitis.

OBJECTIVES/GOALS: The 2020-2021 Galveston County Youth Risk Survey continues past efforts to characterize behavioral risks for local youth, identify disproportionate risks among groups, and provide the data needed for action and intervention to improve the health and safety of our youth in Galveston, Texas. METHODS/STUDY POPULATION: While the survey is based on the CDC Youth Risk Behavior Surveillance System (YRBSS), there was community need for an expanded survey and thus a workgroup was formed to create the Galveston Youth Risk Survey, including members from UTMBs Institute for Translational Sciences, a number of other UTMB departments and Centers, the Research, Education, and Community Health (REACH) Coalition, several school districts, Teen Health Clinic, and a variety of other community health organizations with vested interest. The survey was administered in November 2020 via ScanTron and REDCap to two local high schools with populations greater than 2,000. CDC guidelines for administration of the YRBSS were followed. RESULTS/ANTICIPATED RESULTS: A total of 2,428 students completed the survey from GISD and DISD. The survey provided insight into the following categories, with mental health emerging as a pressing issue, specifically stress and depression, particularly among females. All results are reported in several contexts, including comparisons by gender, grade, and ethnicity, a comparison to state and national statistics when available, as well as risk trends from previous surveys. Demographics Safety, including driving, violence, and bullying Mental Health: stress, depression, suicidal ideation, and ACEs Human Trafficking Substance Use, including smoking, alcohol, marijuana, and other drug use Sexual Behaviors, including contraceptive use Body Weight and Body Image Health Conditions Home Life: Support and Security DISCUSSION/SIGNIFICANCE: The report has been presented to the school districts, immediate stakeholders, REACH membership, and the general public. Several presentations have been given to groups to report the findings. Workgroups will now be formed to address the needs of our students, as well as a possible follow-up survey to look at data specific to COVID-19 and mental health.

OBJECTIVES/GOALS: The objective was to examine the American Heart Association’s (AHA) Essential Eight metrics of cardiovascular (CV) health among Black and White adults with peripheral arterial disease (PAD) collected via validated surveys and medical records. Each metric was examined in association with available social determinants of health (SDoH) factors. METHODS/STUDY POPULATION: This observational study completed data collection through surveys and medical record review. Validated surveys were used to collect Essential Eight metrics of diet, physical activity, sleep, and smoking status. Medical records were used to collect data on body mass index, blood lipids, blood glucose, and blood pressure. Participants with a diagnosis of lower extremity PAD, ability to complete surveys, and provided informed written consent were eligible. Equal numbers of Black and White participants were enrolled. Essential Eight metrics were used to calculate CV health scores for each participant. Scores were examined for association with SDoH factors and by race using Student’s T-test or ANOVA for continuous variables or Chi-Square tests for categorical variables. RESULTS/ANTICIPATED RESULTS: A total of 50 participants will be enrolled, with the expected majority being men and half self-reporting as Black individuals. Worse SDoH is expected to be associated with lower CV health metrics, including lower levels of physical activity and higher levels of saturated fatty food consumption. Higher levels of blood lipids, blood glucose, and blood pressure are expected to be associated with worse SDoH factors. We expect this association to be attenuated by rates of CV medications, such as statin therapy, antiglycemic medications, and antihypertensive medications. No effect modification by rurality is expected, although our projected sample size is small and may impact the ability to examine this interaction. DISCUSSION/SIGNIFICANCE: Black Americans, particularly in the Deep South, are at elevated risk for PAD and critical limb ischemic events, such as amputation. Understanding CV health metrics and SDoH characteristics among adults with PAD is essential to reduce disparities in care and provide valuable information for those at highest risk for complications.

OBJECTIVES/SPECIFIC AIMS: Specialized pro-resolving lipid mediators (SPM) actively counter proinflammatory cascades. A deficit of SPMs is one possible mechanism through which inflammation leads to the development of atherosclerotic disease. The purpose of this study is to characterize the profiles of intermediates of SPM synthesis pathways and end-product SPMs in the plasma of patients with peripheral artery disease (PAD). METHODS/STUDY POPULATION: A cross-sectional sample of 52 patients with PAD was recruited at the San Francisco Veterans Affairs Medical Center. PAD was defined as the presence of claudication symptoms and an ankle-brachial index <0.9, or a history of revascularization for claudication. Patients were excluded if they were taking immunosuppressive medications, had a severe acute illness (infection, surgery, illness, critical limb ischemia) within the last 30 days, or had severe hepatic, renal, or nonvascular inflammatory disease. Intermediates of SPM synthesis pathways and end-product SPMs were measured in plasma samples of patients by liquid chromatography-tandem mass spectrometry. RESULTS/ANTICIPATED RESULTS: The average age of the cohort was 69±6.3 and patient comorbidities reflected common comorbidities associated with PAD (hypertension 96%, hyperlipidemia 87%, diabetes mellitus 42%, coronary artery disease 34%). Rutherford categories, measurements of PAD symptom severity, ranged from 0 to III (0 10%, I 40%, II 27%, III 23%). Three EPA products were measured: 18-hydroxyeicosapentaenoic acid (18-HEPE), resolvin E1 (RvE1), and resolvin E2 (RvE2). 18-HEPE, an intermediate of SPM synthesis, was detectable in the plasma of every patient (median: 105 pg/mL, IQR: 54.9–195), whereas the SPM end-products, RvE1 and RvE2, were only detectable in 6 and 10 patients, respectively. In total, 7 DHA products were measured: 14-hydroxydocosahexaenoic acid (14-HDHA), 17-HDHA, resolvin D1 (RvD1), resolvin D2 (RvD2), protectin D1, protectin DX, and maresin 1. The intermediates 14-HDHA (median: 6546 pg/mL, IQR: 3329–12061) and 17-HDHA (median: 644 pg/mL, IQR: 340–1056) were detectable in the plasma of every patient. However, the end-products RvD1, RvD2, protectin D1, protecin DX, and maresin 1 were identified in less than half of the cohort. DISCUSSION/SIGNIFICANCE OF IMPACT: We report the presence of several intermediates of SPM synthesis pathways (18-HEPE, 14-HDHA, and 17-HDHA) in every patient but the presence of SPM end-products in only a limited portion of the cohort. These results suggest that some patients with PAD may have a deficit of SPMs. Further investigation is required to better understand the role of SPMs and mediators of resolution of inflammation in PAD.

Community engagement is important for reaching vulnerable populations in the coronavirus disease 2019 (COVID-19) pandemic. A risk communication framework was implemented by a community-engaged research (CEnR) partnership in Southeast Minnesota to address COVID-19 prevention, testing, and socioeconomic impacts. Bidirectional communication between Communication Leaders and community members within their social networks was used by the partnership to refine messages, leverage resources, and advise policy makers. Over 14 days, messages were delivered by 24 Communication Leaders in 6 languages across 9 electronic platforms to 9882 individuals within their networks. CEnR partnerships may effectively implement crisis and emergency risk communication to vulnerable populations in a pandemic.

Community-engaged research (CEnR) is now an established research approach. The current research seeks to pilot the systematic and automated identification and categorization of CEnR to facilitate longitudinal tracking using administrative data. We inductively analyzed and manually coded a sample of Institutional Review Board (IRB) protocols. Comparing the variety of partnered relationships in practice with established conceptual classification systems, we developed five categories of partnership: Non-CEnR, Instrumental, Academic-led, Cooperative, and Reciprocal. The coded protocols were used to train a deep-learning algorithm using natural language processing to categorize research. We compared the results to data from three questions added to the IRB application to identify whether studies had a community partner and the type of engagement planned. The preliminary results show that the algorithm is potentially more likely to categorize studies as CEnR compared to investigator-recorded data and to categorize studies at a higher level of engagement. With this approach, universities could use administrative data to inform strategic planning, address progress in meeting community needs, and coordinate efforts across programs and departments. As scholars and technical experts improve the algorithm’s accuracy, universities and research institutions could implement standardized reporting features to track broader trends and accomplishments.

COVID-19 reinforced the need for effective leadership and administration within Clinical and Translational Science Award (CTSA) program hubs in response to a public health crisis. The speed, scale, and persistent evolution of the pandemic forced CTSA hubs to act quickly and remain nimble. The switch to virtual environments paired with supporting program operations, while ensuring the safety and well-being of their team, highlight the critical support role provided by leadership and administration. The pandemic also illustrated the value of emergency planning in supporting organizations’ ability to quickly pivot and adapt. Lessons learned from the pandemic and from other cases of adaptive capacity and preparedness can aid program hubs in promoting and sustaining the overall capabilities of their organizations to prepare for future events.

OBJECTIVES/GOALS: Lung Ultrasound Congestion Score (LUS-CS) is a proposed measure for guiding treatment in acute heart failure (AHF). An emergency department (ED) pilot trial of LUS-guided diuresis showed reduced LUS-CS at 48 hours but no difference at hospital discharge or for clinical outcomes. We hypothesized total change in LUS-CS would predict adverse outcomes. METHODS/STUDY POPULATION: This was a post-hoc secondary analysis of the BLUSHED-AHF trial. BLUSHED-AHF was a pilot trial in which AHF patients were randomized to a LUS-guided diuresis strategy vs. usual care in the ED. The intervention was stopped after the ED course (i.e. during hospitalization). BLUSHED-AHF was designed for the intervention to target absolute values of LUS-CS over time, rather than change in LUS-CS from each patient’s baseline. We fit a cox regression model for a primary outcome of death or AHF rehospitalization, with total (ED to Hospital Discharge) change in LUS-CS as the primary predictor, adjusted for the Get-With-The-Guidelines heart failure risk score (GWTG). Survival curves were plotted, and hazard ratios calculated. RESULTS/ANTICIPATED RESULTS: 128 patients in BLUSHED-AHF were analyzed. Greater reduction in LUS-CS from ED to hospital discharge predicted event-free survival (HR = 0.74 for each 20 unit reduction in LUS-CS, 95%CI 0.56-0.99). This effect did not vary by hospitalization length or ED disposition. There was a significant interaction between change in LUS-CS and GWTG score (p DISCUSSION/SIGNIFICANCE: LUS-CS total change, and not absolute values, predict adverse events in LUS-guided diuresis. Post-ED cessation of the intervention in BLUSHED-AHF may have precluded opportunity for clinical benefit. Future trials should run the entire hospital course, target change from baseline, and consider patient selection by AHF severity and initial LUS-CS.

OBJECTIVES/SPECIFIC AIMS: Recent advances in microneedle technology have enabled practical, in vivo dermal interstitial fluid (ISF) sampling. These minimally-invasive techniques allow for collection of ISF without damage to adjacent tissues and do not rely on blister formation. Initial reports of extracellular vesicle (EV) isolation from dermal ISF and paired blood samples suggest that EVs may be more abundant in ISF. Analysis of ISF-derived EVs may allow for more detailed study of intercellular communication at the tissue level, particularly in acute inflammatory conditions. The objective of this study is to describe the isolation and initial characterization of interstitial fluid-derived exosomes. METHODS/STUDY POPULATION: We apply electron microscopy, nanoparticle tracking analysis (NTA), immuochemical, and sequencing methods to describe and distinguish the EV content of interstitial fluid. We include apparently healthy adult human subjects with no active skin disease. We also study immunocompetent, CD-hairless rats to demonstrate the generalizability of the methods. RESULTS/ANTICIPATED RESULTS: We successfully isolated EVs from human and rat interstitial fluid using commercially available precipitation methods. The EVs were initially characterized using UV/Vis spectroscopy, electron microscopy, and NTA. While the study is ongoing, initial results suggest that the concentration and size distribution of EVs differs significantly between blood fractions and ISF. Further immunochemical and sequencing characterization is ongoing. DISCUSSION/SIGNIFICANCE OF IMPACT: We present here the initial characterization of EVs isolated from dermal interstitial fluid. This appears to be the first report of EV characterization using ISF collection methods that do not perturb adjacent tissues (such as with blister or microdialysis methods). The present study lays a foundation for further examination of ISF-derived EVs in acute inflammatory disease such as cellulitis or infectious neuritis. This may enable minimally invasive diagnostics and new research tools to understand intercellular communication in living organisms with increased spatial and temporal resolution.

OBJECTIVES/GOALS: Weight stigma, or discrimination and devaluation of people due to high body weight, is associated with higher prevalence of concerning health consequences (e.g., depressive symptoms, body dissatisfaction). This public health research investigated how experiencing weight stigma affects weight-related health outcomes in a cohort of young people. METHODS/STUDY POPULATION: 1,534 Project EAT 2010-2018 participants were surveyed as adolescents (mean age=14.4 years) and eight years later as young adults (mean age=22.2 years). Participants were asked how often they were teased about their weight. Outcomes included body mass index (BMI), nutrition habits (intake of breakfast, fruit, vegetables, sugar-sweetened beverages, fast-food), sedentary/activity behaviors (moderate-to-vigorous physical activity, screen time, sleep duration), and disordered eating behaviors (unhealthy weight control behaviors, extreme weight control behaviors, dieting, chronic dieting, overeating, binge eating). Regression models were adjusted for BMI, sociodemographic characteristics, and, in longitudinal models, the outcome assessed at baseline. RESULTS/ANTICIPATED RESULTS: Experiencing weight stigma was cross-sectionally significantly associated with higher BMI, shorter sleep duration, and higher prevalence of all six disordered eating behaviors during both adolescence and young adulthood. It was also significantly associated with longer screen time in adolescence, and with lower breakfast frequency, higher intake of sugar-sweetened beverages, and higher intake of fast-food during young adulthood. Weight stigma in adolescence significantly predicted higher BMI and higher prevalence of overeating and dieting in young adulthood. For example, adolescents who were teased about their weight had a higher mean BMI (28.2 kg/m2 [95% confidence interval: 27.8-28.7]) in young adulthood compared to those who had not been teased (26.4 kg/m2 [95% confidence interval: 26.1-26.8]). DISCUSSION/SIGNIFICANCE: Findings add to the growing body of evidence that weight stigma may pose a threat to weight-related health. Because many clinicians are unaware of this evidence and healthcare settings are common sources of weight stigma, next steps include writing a research proposal that aims to decrease weight stigma conveyed to patients in clinical practice.

OBJECTIVES/SPECIFIC AIMS: Our objective is to study the role of soluble adenylyl cyclase in the melanocyte regulation of pigment in response to ultraviolet radiation. Melanocytes are specialized cells that produce melanin in organelles called melanosomes, and melanin determines the pigmentation of hair and skin. cAMP is a master regulator of pigmentation and transmembrane class of adenylyl cyclases are essential for expression of important enzymes involved in melanogenesis. However, pigmentation is also controlled by melanosomal pH, which regulates melanogenesis, tyrosinase activity, and melanosome maturation. The relationship between melanosomal pH and cAMP has been elusive. Soluble adenylyl cyclase is a noncanonical source of cAMP that is not responsive to G proteins but rather functions as a pH sensor. We recently demonstrated that loss of soluble adenylyl cyclase (sAC) activity leads to increased melanosomal pH as well as increased pigmentation in cells and hair. We expanded our research to investigate the role of sAC in the intrinsic response of melanocytes to ultraviolet radiation. METHODS/STUDY POPULATION: We utilized sACfl/fl (wild type) and sACKO mouse melanocytes and compared their change in pigmentation in response to ultraviolet radiation. Melanin was used as a measure of pigmentation. We irradiated these cells at differing doses of UVB (0, 1, 2, or 3 mJ/cm2) daily for 3 days. After UVB treatment, cells were observed and the surviving cell numbers were determined. Cells were then analyzed for melanin content using spectroscopy. RESULTS/ANTICIPATED RESULTS: We found that while both sACfl/fl and sACKO cells had increased melanin content in response to UVB, the melanin content of sACKO cells increased more compared with sACfl/fl cells (p=0.001 at daily dose of 3 mJ/cm2). In addition, sACKO cells required less UVB dose to induce a response. We also observed that sACKO cells show increased cell death compared with sACfl/fl cells. DISCUSSION/SIGNIFICANCE OF IMPACT: Although both sACfl/fl and sACKO cells can induce melanin production in response to UV, our results suggest that sACKO cells are more sensitive. We believe that this increased response in sACKO cells is due to increased melanosomal pH. In addition, sACKO cells show increased cell death, suggesting that sAC is important in the damage response secondary to UV exposure. UV plays a wide range of roles in skin biology such as contributing to cancer risk and pigmentation. Since pigmentation is essential for the protection of the skin from UV insult, further investigation of possible mechanisms in which sAC can influence pigmentation in response to UV is warranted.

OBJECTIVES/GOALS: We developed a non-invasive test of ocular alignment (OA) as a measure of otolith asymmetry, which impacts vestibular function. This test has also been administered on commercial astronauts to study the effect of microgravity on vestibular function. We will assess OA test performance in ground subjects, which has not yet been characterized. METHODS/STUDY POPULATION: Subjects were recruited from visitors to Space Center Houston from June to October 2022. Participants wore red-blue glasses in a darkened room and aligned two vertically or torsionally misaligned line segments, one red and one blue, on a tablet. Each subject underwent 11 vertical and torsional trials. The remaining misalignment was used to calculate ocular misalignment. Subjects self-reported demographic and health data. We calculated descriptive statistics and explored the distribution of overall vertical and torsional OA and distributions by age. Next steps will establish the reliability of the outcome measures with intraclass correlation coefficients and analyze associations between demographic variables and health status with OA performance using linear regression analysis. RESULTS/ANTICIPATED RESULTS: A total of 143 participants (67 female) with a median age of 21 (range 5-74 years old) completed OA testing. Of participants, 64% identified as white, 10% as black, 14% as Asian, 3% as Native American, and 8% as unknown. This test battery was feasible to implement in this mixed-age sample, with a 95% completion rate. Median absolute vertical OA was 0.08 degrees (IQR = 0.17 degrees), and median absolute torsional OA was 0.80 degrees (IQR = 1.20 degrees). Visual analysis of plots of OA by trial number reveals no apparent learning effect within subjects, though some individuals had considerable variation in performance. Average absolute vertical ocular misalignment increased modestly when plotted by age (fitted regression line b = 0.0032 degrees per year of age, p = 0.004), unadjusted for potential confounders. DISCUSSION/SIGNIFICANCE: Ocular alignment testing was feasible in a mixed-age general population sample. Our findings will expand understanding of how OA varies with age. These results will be used to create a normative database to compare and inform analysis of OA results from commercial spaceflights, which include individuals of varying age and health statuses.

OBJECTIVES/SPECIFIC AIMS: Our research strives to understand the pathophysiology of doxorubicin cardiotoxicity, focusing on the understudied nonmyocyte cardiac cells. Our understanding will enable researchers to develop protective or alternative therapies for cancer patients and treatments for cancer survivors. METHODS/STUDY POPULATION: Early studies have been carried out in isolated primary cardiac fibroblasts. Cells were treated with varying doses of doxorubicin. Cell viability, proliferation, and reactive oxygen species generation have all been studied. Future studies will focus on mitochondrial assessment in treated cells and confirmation of findings in animal models. Potential therapies discovered in these studies will also be conducted in animal models. RESULTS/ANTICIPATED RESULTS: Our results show a direct effect of doxorubicin on cardiac fibroblasts in vitro. Treated cells show a decreased rate of proliferation and increased production of reactive oxygen species. Similarly to cardiomyocytes, we hypothesize that reactive oxygen species damage the mitochondria of cardiac fibroblasts thereby altering their function and playing a role in doxorubicin cardiotoxicity. DISCUSSION/SIGNIFICANCE OF IMPACT: Current therapies have not been able to adequately protect patients from the cardiotoxicity of doxorubicin and other anthracyclines. A complete understanding of how doxorubicin damages cardiac tissue will only be possible by studying all cell types of the heart. With a better understanding, alternative therapies can be developed to prevent or treat doxorubicin cardiotoxicity without sacrificing the efficacy of doxorubicin in treating cancer.

OBJECTIVES/GOALS: Single nucleotide polymorphisms in the transcription factor 7-like 2 (TCF7L2) gene are associated with Type 2 Diabetes (T2D) and nonalcoholic fatty liver disease (NAFLD). The metabolic function of TCF7L2 in the liver remains to be fully elucidated, but we hypothesized that TCF7L2 contributes to NAFLD through regulation of zonal metabolic pathways. METHODS/STUDY POPULATION: Using single nuclei RNA sequencing, we examined Tcf7l2 expression in periportal (PP) hepatocytes around the portal triad and pericentral (PC) hepatocytes surrounding the central vein of the liver. To visualize TCF7L2 transcriptional activity we used a TCF reporter mice, which expresses an H2B-eGFP fusion protein downstream of the conserved TCF DNA binding site. We disrupted Tcf7l2 transcriptional activity in mouse liver by breeding mice with a floxed Tcf7l2 exon 11, which encodes part of the DNA binding domain (DBD), to albumin-Cre mice (Hep-TCF7L2ΔDBD). Eight-week-old mice were fed a choline-deficient amino acid-defined high fat (CDAHFD) diet for 8 weeks. In liver samples harvested from these mice, we examined disruption to several key zonated metabolic pathways, and quantified the development of fibrosis. RESULTS/ANTICIPATED RESULTS: Single nuclei analysis revealed that Tcf7l2 mRNA was expressed primarily in parenchymal cells of the liver but was ubiquitous across the liver lobule. However, in immunofluorescence analysis of TCF reporter mice, the transcriptional activity of TCF7L2 was highly restricted to PC hepatocytes. Classic PC hepatocyte markers, including glutamine synthetase (Glul), were absent in Hep-TCF7L2ΔDBD mice. Following the CDAHFD, Hep-TCF7L2ΔDBD mice developed more severe fibrosis in histological analysis, and expressed elevated levels of genes involved in fibrogenesis, collagen synthesis and TGFβ signaling. Hep-TCF7L2ΔDBD mice also displayed hepatic cholesterol accumulation following the CDAHFD, which was likely the result of impaired pericentral bile acid synthesis. DISCUSSION/SIGNIFICANCE: Our results suggest that TCF7L2 plays an important role in the regulation of zonated metabolic pathways, which may contribute to the development of fibrosis. Ongoing analyses are exploring the mechanisms regulating the zonal transcriptional activity of TCF7L2.

OBJECTIVES/GOALS: Pancreatic ductal adenocarcinoma (PDAC) is projected to become the second leading cause of cancer-related deaths by 2030. Though many other cancers have seen improvements in patient survival rates, patients diagnosed with PDAC have a 5-year survival rate of only ~9%. A major contributor to decreased survival is late-stage diagnosis of the disease. New methods of early detection are urgently needed. Extracellular vesicles (EVs) are secreted from cells of all tissue types into the circulation. EVs play important roles in a variety of diseases. They have shown to promote cancer progression and they are being studied as potential biomarkers for disease diagnosis. The purpose of this study was to perform qualitative and quantitative characterization of small-molecule profiles of EVs derived from various pancreatic cancer (PC) and normal pancreas cell lines, to provide proof-of-concept for evaluating the efficacy of leveraging EVs as potential biomarkers of PDAC. METHODS/STUDY POPULATION: EVs were isolated from the conditioned media of six PC and two normal pancreas cell lines using differential ultracentrifugation with filtration. EV enrichment was validated using quantitative ELISA, immunoblot and transmission electron microscopy. Targeted liquid chromatography coupled to mass spectrometry (LC-MS/MS) and untargeted (UPLC-QTOF-MS) metabolomics were used to analyze the biochemical composition of EVs. RESULTS/ANTICIPATED RESULTS: The biochemical profile of PC EVs was found to be significantly different from the profiles of normal cell EVs. Interestingly, amino acids were downregulated in PC EVs as compared to normal cell EVs. However, PC EVs were enriched in lactate and malate. PC EVs also had significant upregulation in other small molecules such as xanthosine, guanosine diphosphate and nicotinamide. DISCUSSION/SIGNIFICANCE OF IMPACT: Our results indicate that the biochemical characterization of EVs using metabolomics has the potential to yield biomarkers which can delineate cancer cell-derived EVs from normal cell-derived EVs. Further work will test the clinical significance of these findings by similar analyses of plasma of PDAC patients. Furthermore, these profiles may be detectable before progression of the disease to late-stage PDAC, leading to the development of assays for earlier diagnosis in patients.

ABSTRACT IMPACT: The Bench Tutorials Program is an independent study course in biomedical research in which high school students are paired with graduate and post-doctoral students during the academic year. The purpose is to enhance the rigor of high school science education and build the pipeline of tomorrow’s researchers. OBJECTIVES/GOALS: The Bench Tutorials Program: ο Proficiency in research design, implementation, and presentation; ο Acquisition of hands-on laboratory skills; ο Increase in scientific literacy; ο Increase in analytical skills and critical thinking; ο Career in science; ο Build the pipeline of tomorrow’s biomedical researchers METHODS/STUDY POPULATION: High School seniors are paired with graduate and postdoc mentors through a matching process. Students spend approximately four hours/week in supervised instruction and research from a participating laboratory in addition to classroom experience at their High School. Mentors design research projects relating to the larger research framework of their laboratories. In light of COVID-19, approaches have been adjusted to maintain the program safely through a hybrid method of using the high school lab for hands-on learning and through the use of Go-Pros ’s to enable our mentors to video and narrate as they conduct experiments in their own labs to teach their mentees scientific methods and processes. RESULTS/ANTICIPATED RESULTS: Since inception, more than 400 students and mentors have participated in the Bench Tutorial’s program. This year we found a way to continue the program under COVID-19 restraints without putting anyone in harms way. Go-Pros have been essential for our program to maintain continuity for high school students who receive academic credit for this course. This program is also one of few in which our graduate students have the opportunity to serve as mentors in the scientific setting. Using Go-Pro’s will also enable us to provide teaching videos online for other academic institutions, so even in the absence of COVID-19 in the future, the continued use of these devices will still be of great value. DISCUSSION/SIGNIFICANCE OF FINDINGS: High school students are afforded the ability to work on cutting edge research projects alongside graduate students and postdocs, who are afforded the chance to mentor and teach. Due to the COVID-19 pandemic, we have successfully adjusted our methods for teaching through the use of Go-Pro technology.

OBJECTIVES/GOALS: High serum copper (Cu) levels have previously been described in bariatric patients. The kidneys are a target organ for Cu toxic insult but the role of Cu on kidney function (eGFR) is uncertain. This study examines the association between Cu and eGFR in a bariatric population in Southeast Louisiana. METHODS/STUDY POPULATION: Seven hundred fifty patients will be recruited from the Bariatric Center of the University Medical Center in New Orleans. Inclusion criteria include: age ≥ 18 years, clinic visit between June 1, 2018 – May 31st 2024, and having a serum Cu test result. Covariables such as inflammatory markers and hormonal contraception use will be assessed as potential confounders. Blood pressure will be assessed as a potential effect modifier. Data will be obtained from electronic medical records. Two cohorts will be assembled, a pre-surgery cross-sectional cohort and another followed post-surgery. Separate models will be developed stratified by race-ethnicity. RESULTS/ANTICIPATED RESULTS: In a pilot study of bariatric patients 26% had elevated (>155 mcg/dl) serum Cu and pronounced racial differences were noted. Characteristics consisted of a mean BMI of approximately 50 kg/m2; 91% were female and 69% were Black. Black patients had approximately double the prevalence (OR 1.98; 95% CI: 1.15, 3.4) compared to white patients. Due to the dual nature of the kidneys’ involvement in metabolism via excretion and being the target organ for toxic insult, racial differences in exposure, coupled with the disproportionate rates of chronic kidney disease in Black adults, may be an explanation for the association between elevated Cu levels and eGFR in Black adults in this study. DISCUSSION/SIGNIFICANCE: Results from this study will provide insight into the prevalence of Cu and its association with kidney function in a bariatric population. Chronic kidney disease or other forms of renal impairment may result in the need for more conservative guidelines for dietary copper in bariatric medicine.