OBJECTIVES/GOALS: A UCLA Clinical and Translational Science Institute (CTSI) science team partnered with the second largest US school district, with over 500,000 K-12 students, to design and implement a statistical process control dashboard to guide COVID-19 response, including mitigation and vaccination outreach. METHODS/STUDY POPULATION: District data for students, teachers, and staff are updated daily and include COVID-19 test results, counts of quarantine after positive tests, and COVID-19 vaccination rates. Displays used a new hybrid Shewhart control chart to detect changes in test positivity rates and distinguish meaningful signals from noise (random day-to-day variation). The dashboard uses the Shiny and plotly packages in R to display interactive graphs of each data stream (cases, tests, and vaccinations) charted at multiple levels (districtwide, subdistricts, schools). Displays of variation over time show policy impacts and inequities. Selected displays use municipal COVID-19 data to complement district data. RESULTS/ANTICIPATED RESULTS: The district has used the displays to assess the impact of their COVID-19 response and to identify variation in close to real-time to suggest areas with need for additional resources for mitigation or vaccination. The CTSI team has continued to edit and add displays in response to the district’s changing operational needs and questions. DISCUSSION/SIGNIFICANCE: The UCLA CTSI team developed and implemented a robust data visualization dashboard to monitor COVID-19 case rates and plan vaccination outreach efforts. Control charts enabled the district to distinguish noise from signal, thereby rapidly identifying when specific parts of the district needed targeted support to achieve equity goals.

Objectives/Goals: Retention in care is vital for people living with HIV. We used human-centered design (HCD) to engage a community-based research panel over a 5-year period, allowing us to incorporate their insights on research guidance and interpretation of findings to investigate correlates of HIV care outcomes. Methods/Study Population: We recruited a diverse panel of individuals who were living with HIV, HIV clinicians, and/or providing non-clinical HIV services in Marion County, Indiana. We conducted biannual sessions using a variety of HCD tools and activities to engage participants. Each session took about three hours, and panelists were compensated for their participation. Due to the COVID-19 pandemic, sessions were initially held virtually. Sessions were designed for project discussion and to facilitate exploration of concerns and challenges facing receipt of HIV services. Our HCD approach put participants in the center of discussion and empowered them to externalize ideas and collaborate meaningfully with our team. Results/Anticipated Results: Since project inception, 48 individuals have joined the panel. Thirty-five are actively engaged, participating in one or more of six sessions conducted to date. We have learned much from the panel. One example is that a residential move might be a risk or protective factor for retention in care and the amount of time one had lived with HIV is a crucial factor. Panel insights have helped guide and prioritize analyses, aided in identification of data missing from our ecosystem, helped interpret results, provided feedback on future interventions, led to a quality improvement project with the local health department, and led to a presentation at a local health equity conference. Discussion/Significance of Impact: Community engagement is essential to impactful and sustainable research. HCD was a successful approach to engage our panel to inform interventions more relevant to the community. We anticipate these methods will be important for others conducting community-engaged research.

OBJECTIVES/GOALS: The goal of this study is to leverage a national database to see if autologous reconstruction rates differ in patient and clinical characteristics, readmission rates, and overall survival (OS) compared to other forms of reconstruction. Autologous reconstruction has not been looked at in this way before. METHODS/STUDY POPULATION: • Aim 1: Use the National Cancer Data Base to construct three patient cohorts for women under 70 and above 18 treated surgically for breast cancer with A) mastectomy only, B) implant-based reconstruction, and C) autologous breast reconstruction. • Aim 2: Examine receipt rates of surgical intervention in Cohorts A vs. B vs. C based on clinical and patient demographic/socioeconomic characteristics. • Aim 3: Compare readmission and overall survival (OS) rates for Cohorts A vs. B vs. C while controlling for age and other key variables. RESULTS/ANTICIPATED RESULTS: Based on the literature, we expect rates of autologous reconstruction (Cohort C) to be lower for patients of minority backgrounds compared to white individuals. In addition, we do not expect overall survival to differ between implant-based (Cohort B) and Cohort C reconstruction. Still, we expect mastectomy-only (Cohort A) survival to vary from the two cohorts even when adjusting for different clinical factors, as similar but smaller studies have shown. Finally, we expect readmission rates to be higher for Cohort C, compared to Cohorts A & B, as it is a more complicated procedure typically done in academic institutions with skilled surgeons. DISCUSSION/SIGNIFICANCE: Autologous reconstruction is now considered the gold standard due to its ability to restore the breast shape with higher patient satisfaction and superior long-term outcomes. Multiple studies have documented ongoing racial disparities in post-mastectomy breast reconstruction and autologous reconstruction, with lower rates and referrals.

OBJECTIVES/SPECIFIC AIMS: To identify cardiac structural and function parameters, obtained on usual stroke-care TTE evaluation, associated with cardioembolic stroke (CE) in patients without AF. Hypothesis—left atrial (LA) size and valve dysfunction will be strongly associated with incident CE. METHODS/STUDY POPULATION: Inclusion criteria: July 1, 2013 to July 1, 2015 admission with imaging-confirmed ischemic stroke, no AF, TTE within 1st 7 days. TTE structure/function parameters were recorded. Stroke subtype (CE vs. other) defined using TOAST criteria, blinded to TTE. New AF definition: AF on ECG, telemetry or event monitor. CE/New AF outcome of interest in separate multivariable logistic regression models testing associations with TTE parameters (adjusting for demographics/vascular risk factors). RESULTS/ANTICIPATED RESULTS: Participants (n=332) were ~60 years hypertensive black males with moderate NIHSS and normal ejection fraction. In adjusted models, odds of CE increased with increasing LA systolic diameter (per 0.1 cm), mitral E point velocity(cm/s), mitral valve dysfunction, wall motion abnormality. New AF also associated with increasing LA systolic diameter. DISCUSSION/SIGNIFICANCE OF IMPACT: These findings may suggest cardiac structural changes independent of AF that are on the CE causal pathway. Understanding the relationship between such TTE parameters and stroke subtype would impact clinical practice, as such TTE data is underutilized when considering stroke mechanism and management.

Objectives/Goals: To describe the pathway of support trial-care has put into place for washu and partner ctsa researchers navigating the investigator-initiated clinical trial life cycle. Methods/Study Population: Trial-care is a centralized research support service at washu supported by the center for clinical studies (ccs) and the institute of clinical and translational sciences (icts). our team consists of experienced investigators and research professionals with expertise in clinical trial management across all phases of the trial life cycle. for washu and partner ctsa researchers planning investigator-initiated clinical trials, we provide a free 60-minute consultation to analyze barriers and streamline processes. after the initial consultation, researchers may receive trial-care services (e.g., project management, quality and regulatory management, and/or data management). these services can be billed at an hourly rate or through direct sourcing of trial-care staff to the funded grant. Results/Anticipated Results: Trial-CARE’s support for investigator-initiated clinical trials accelerates all phases of the trial life cycle, such as pre-award planning and post-award start-up, implementation, and closeout. Since 2019, Trial-CARE has completed over 142 consultations across 20 Departments at WashU, SLU, and Mizzou. At the time of consult, 54.4% were in the pre-award planning phase and 45.6% were in the post-award start-up, implementation, or closeout phase. About 20% of consultations lead to the utility of Trial-CARE Services, and the top reasons researchers connect for consultation to receive Trial-CARE Services: (1) regulatory guidance (FDA Part 11, IND, IDE, IRB) = 55% (2) safety monitoring or site/data monitoring = 33% (3) data management = 23% (4) study budgeting = 19% (5) protocol development = 17%. Discussion/Significance of Impact: Trial-care uses a clinical trial dashboard to identify and track pre-award investigator-initiated clinical trials and is developing outreach and training programs to pro-actively engage these studies. the goal is to maximize clinical trial impact through quality (rigor and best practices), innovative capacity, and reproducibility.

OBJECTIVES/GOALS: Individuals with a family history of alcoholism (FH+) are more likely to develop an alcohol use disorder than those with no such history. Early life adversity has a high coincidence with FH+ making pathogenic studies difficult in clinical studies. Here, we developed a mouse model to study pathogenic mechanisms underlying these risk factors. METHODS/STUDY POPULATION: Male and female C57BL6/J mice were exposed to increasing concentrations of ethanol (3-21%) or water for 15 days prior to breeding. Ethanol was not present during gestation. Offspring were either removed from the home cage and isolated for 3 hours or left undisturbed from postnatal days 1-21. Beginning on PND 56 offspring mice were assessed for clinically relevant behavioral disruptions in social behavior, cognitive working memory, locomotor activity, anxiety-like phenotypes, ethanol preference and binge drinking behavior. In a separate experiment, brains of Cx3cr2+/GFPxCcr2+/RFP mice from ELA or control conditions were collected every 7 days after birth for assessment of neuroinflammation and central immune cell morphology and density. RESULTS/ANTICIPATED RESULTS: Mice with a family history of ethanol exposure and ELA are predicted to exhibit behavioral changes (impaired working memory, reduced social behavior, increased anxiety-like behaviors, increased ethanol consumption) to a greater extent than mice with a family history of ethanol exposure or ELA alone. We expect markers of neuroinflammation (cytokine expression, immune cell activation) to predict the behavioral changes in these mice. DISCUSSION/SIGNIFICANCE OF IMPACT: Alcohol consumption and stressful life events are known environmental precipitants to neuroinflammation, which in turns may predispose individuals to anti-social and risky behavior. A mouse model of these early postnatal conditions will allow basic scientists to unravel the biological underpinnings of the behaviors driven by these factors.

ABSTRACT IMPACT: Our improved understanding of the changes in chromatin accessibility that occur in persistent pain states may identify regulatory genomic elements that play essential roles in modulating gene expression in the DRG. OBJECTIVES/GOALS: Efforts to understand genetic variability involved in an individual’s susceptibility to persistent pain support a role for upstream regulation by epigenetic mechanisms. Our objective was to examine the transcriptomic and epigenetic basis of persistent pain following nerve injury. METHODS/STUDY POPULATION: We used a multiomic approach to identify novel molecular pathways associated with nerve injury-induced pain hypersensitivity. Adult Sprague Dawley rats were randomized to Chronic Constriction Injury (CCI) to the sciatic nerve or no treatment (naive). The ipsilateral L4-L6 dorsal root ganglia (DRG)s were removed on Day 14 and used for ChIP-seq for H3K4me1, ATAC-seq, and RNA-seq. We assessed for differential chromatin accessibility, transcription factor motifs, and enrichment for biological processes in chromatin accessible regions associated with cis-regulatory regions identified by ATAC-seq and H3K4me1 enrichment. Luciferase assays determined the functional significance of these sequences. RESULTS/ANTICIPATED RESULTS: We identified 58,446 genomic regions where H3K4me1 enrichment overlapped with chromatin accessibility. Differential analysis identified 2145 of these 58,446 regions that had changes in accessibility after CCI. The majority of these regions were located in introns or intergenic regions. Functional annotation of the differentially accessible regions identified disparate molecular functions enriched following nerve injury which suggests that altered chromatin structure plays a role in the development of mechanical hypersensitivity. Motif analysis identified specific transcription factor families whose binding sequences were enriched in regions of increased or decreased accessibility. Luciferase assays showed significant enhancement or repression of gene transcription. DISCUSSION/SIGNIFICANCE OF FINDINGS: Our data provides a comprehensive map of chromatin accessibility changes in the DRG after CCI and emphasizes the importance of chromatin structure in the development and maintenance of chronic pain.

OBJECTIVES/SPECIFIC AIMS: Hematopoietic stem and progenitor cells (HSPCs) function to maintain steady state production of new blood cells and to rapidly respond to blood cell loss. Little is known regarding how HSPCs develop the ability to sense and respond to blood cell loss during embryogenesis. Gaining insight into the robust ability of HSPCs to regenerate blood during early development may allow us to develop therapies to rejuvenate this capacity at any stage. METHODS/STUDY POPULATION: We generated a new hematopoietic-specific and inducible cell ablation zebrafish model to uncover the origins of regenerative capacity in HSPCs during development. These transgenic zebrafish express a cyan fluorescent protein (CFP)-nitroreductase (NTR) fusion construct under the control of the draculin (drl) promoter (drl:CFP-NTR), which restricts NTR expression to blood cells. Co-expression analyses of drl:CFP-NTR with known markers of other blood types including HSPCs (runx1+23:mCherry), erythroid cells (gata1:dsRed), and lymphoid cells (rag2:RFP), revealed drl:CFP-NTR was restricted to HSPCs and erythrocytes. To delineate the regeneration potential of embryonic HSPCs, we exposed drl:CFP-NTR transgenic zebrafish embryos to Metronidazole (MTZ), which results in selective ablation of only NTR-expressing blood cells. Embryos were treated from 2 to 3 days postfertilization and recovery of drl+ and gata1+ cells was evaluated over a 7-day recovery period. RESULTS/ANTICIPATED RESULTS: Following MTZ exposure, the nadir of drl+ cell ablation occurs at 2 days post MTZ (dpM) treatment. During the renewal phase of blood regeneration, we first observe recovery of drl+ cells by about 4 dpM, while more mature blood cells like gata1+ erythrocytes show a delayed recovery at about 6 dpM. Our results suggest that HSPCs can respond to injury as early as 2 days of life and that the HSPC-driven regeneration of embryonic blood cells occurs in a hierarchical fashion, similar to regeneration of the adult blood system. DISCUSSION/SIGNIFICANCE OF IMPACT: We have established a quantitative method for in vivo real-time monitoring of embryonic and larval blood regeneration. A significant advantage of our system is that we can use these insights to guide an in-vivo drug screen for factors that accelerate HSPC-driven blood regeneration in a complex biological environment.

OBJECTIVES/GOALS: For shoulder osteoarthritis (OA), the understanding of the patient-specific factors that determine success of both non-operative and operative treatment options is limited. This study aims to identify key factors associated with the response and the heterogeneity of outcomes for both types of treatment. METHODS/STUDY POPULATION: Patients diagnosed with shoulder OA and treated with either reverse/anatomic total shoulder arthroplasty (rTSA/TSA) or non-operative management at the University of California, San Francisco were enrolled in this study. They were followed for a year to ascertain phenotypic traits and patient-reported outcomes (PROs). Magnetic Resonance Imaging (MRI) was used to calculate the Shoulder Osteoarthritis Severity (SOAS) score, a semi-quantitative global assessment of shoulder OA, and to measure fat fractions of rotator cuff muscles. A Microsoft Kinect camera was used to determine the Reachable Workspace (RWS). Linear regression models were used to assess the associations between baseline demographic and radiographic factors on outcomes related to shoulder function. RESULTS/ANTICIPATED RESULTS: It is anticipated that the pre-operative MRI-based SOAS score will be inversely correlated with the magnitude of improvement in PROs 1 year after rTSA/TSA and non-operative management of shoulder OA for the surgical replacement and non-operative cohorts, respectively. Additionally, the non-operative patients who convert to rTSA/TSA within 1 year of observation will have higher SOAS scores compared to patients who continue with non-operative management. The surgical replacement patients with an infraspinatus fat fraction of more than 5% will have worse shoulder function, as measured by RWS, compared to patients with an infraspinatus fraction less than 5%. DISCUSSION/SIGNIFICANCE: MRI may be a novel technique to better predict prognosis of shoulder OA management. This will allow for the development of appropriate algorithms in the prescription of treatments and may be used to counsel patients regarding their expected outcomes or to recommend alternative treatments.

OBJECTIVES/GOALS: The primary objective is to measure the independent association of hospital-level and sociodemographic variables on the rate of hospital-onset bacteremia among infants admitted to the neonatal intensive care unit in a United States of America retrospective cohort. The secondary outcome will be relative blood culture collection rate. METHODS/STUDY POPULATION: The study is an analysis of a retrospective cohort comprised of infants admitted to 322 neonatal intensive care units (NICUs) in the United States of America between 2016-2021. The primary outcome will be hospital-onset bacteremia (HOB), defined as a positive blood culture with a bacteria or fungi after day 3 of admission. Independent risk factors will include birthweight, postnatal age, central venous catheter presence, sociodemographic variables (race, ethnicity, insurance status and ZIP code-level demographic data from the US Census American Community Survey (ACS), and hospital-level variables. Infants will be stratified by sociodemographic groups and a Poisson model will be utilized to measure the adjusted association between risk of HOB and clinical and hospital-level variables. RESULTS/ANTICIPATED RESULTS: I anticipate that infants in sociodemographic groups with a history of socioeconomic marginalization will have a higher unadjusted rate of HOB; however, sociodemographic variables will not be independently associated with HOB risk after adjusting for markers of hospital quality and acuity, such as quartiles of the following: mean admissions per year, percentage of infants born <1500g, annual blood culture contamination rate, and percentage infants born at another facility. DISCUSSION/SIGNIFICANCE: Neonatal bacteremia has high morbidity and mortality; however, its contribution to known infant mortality inequities is unknown. This study will estimate the burden of infant HOB stratified by sociodemographic groups and measure the independent association of sociodemographic and hospital-level variables on the adjusted rate of HOB.

OBJECTIVES/GOALS: To describe and compare clinical data and outcomes for patients with CNS tumors and tumor mimics in Puerto Rico who are undergoing surgical and nonsurgical management. Thus, increasing data from an underrepresented group which can serve as a foundation for investigating determinants of outcomes. METHODS/STUDY POPULATION: This proposal will examine patient charts, radiology and pathology reports, financial data, and treatment details from the electronic medical record of patients receiving surgical and nonsurgical treatment for CNS tumors and mimics in the University of Puerto Rico Medical Sciences Campus and all associated institutions. Data will be analyzed retrospectively between January 1, 2014 and June 30, 2022, and prospectively for ten years until December 31, 2032. Patients with primary and metastatic CNS tumors and tumor mimics in the brain, meninges, ventricles, spinal cord, cranial nerves, orbit, facial sinuses, bony skull, vasculature will be included. The registry will include patients from birth onward. RESULTS/ANTICIPATED RESULTS: We plan to compare different surgical and non-surgical techniques and devices in terms of technical and clinical outcomes after surgical interventions for CNS tumors. We are collaborating with the CNS Tumor Outcome Registry at Emory (CTORE) and plan to continue collaboration with other institutions. Combining our data, we aim to develop predictive models of patient outcomes after surgical and nonsurgical intervention for CNS pathologies using supervised and unsupervised machine learning strategies. DISCUSSION/SIGNIFICANCE: There is a significant lack of literature on CNS intervention outcomes in Puerto Rico. This registry will provide the platform for cost-analysis studies for techniques and clinical protocols applicable to pre-operative, intra-operative, post-operative, and conservative management of patients, in Puerto Rico and beyond.

ABSTRACT IMPACT: Understanding the biology underpinning head and neck cancer invasion and metastasis could lead to novel targeted therapies that are both effective and tolerable for patients with this debilitating disease. OBJECTIVES/GOALS: The bromodomain and extra-terminal (BET) family of epigenetic regulators has been implicated in the tumorigenesis of various cancers. In head and neck squamous cell carcinoma (HNSCC), the majority of morbidity is due to invasion and metastasis, so there is special interest in understanding the development of these phenotypes in HNSCC cells. METHODS/STUDY POPULATION: Stable SCC9 knockdown cell lines were generated by infecting cells with a lentivirus encoding Cas9-KRAB and a lentivirus encoding gRNA targeting each of the candidate BET proteins: BRD2, BRD3, BRD4, and BRDT. Knockdown was confirmed by qRT-PCR. Next, standard assays for proliferation (CellTiter-glo), invasion (Matrigel), and migration (scratch-wound healing assay) were performed for all candidate knockdowns and compared to a non-target control. RESULTS/ANTICIPATED RESULTS: Proliferation assay results revealed that BRD4 knockdown had a significant negative effect on the proliferative capacity of SCC9 cells in vitro. Similarly, BRD4 knockdown SCC9 cell lines were less invasive and less migratory. Interestingly, knockdown of BRD2, BRD3, and BRDT had no effect on proliferation, invasiveness, or migration. DISCUSSION/SIGNIFICANCE OF FINDINGS: We have identified BRD4 as a key driver of the HNSCC tumorigenic phenotype. In the future, we plan to investigate the role of JQ1, a pan-BET inhibitor, on HNSCC cell phenotypes. Additionally, we will identify the downstream targets of BRD4, which may serve as potential therapeutic targets for both HNSCC as well as other cancers more broadly.

OBJECTIVES/SPECIFIC AIMS: Drug repositioning has the potential to accelerate translation of novel cancer chemotherapeutics from bench to bedside. The goal of this study was to determine the effects of ciclopirox olamine (CPX) on esophageal tumor cells. METHODS/STUDY POPULATION: We tested the effect of CPX on four esophageal cancer cell lines, assessing cell proliferation and viability by hexosaminidase and clonogenicity assay, respectively. We analyzed the effects of CPX on three-dimensional (3D) esophageal tumor cell spheroids. We also analyzed effects on cell cycle by flow cytometry. For mechanism, we performed western blots for proteins involved in cell cycle regulation, apoptosis and the Wnt/β-catenin pathway. For in vivo effects, we performed a murine xenograft model with intraperitoneal administration of CPX (100 mg/Kg body weight daily). RESULTS/ANTICIPATED RESULTS: CPX inhibited growth of all cell lines in a time and concentration-dependent manner. CPX also inhibited growth of esophageal spheroids. Cell cycle analysis demonstrated G0/G1 arrest in cells treated with CPX. Western blot analyses demonstrated decreased expression of cyclinD1, CDK4, CDK6, and transcriptionally active β-catenin, supporting the role of CPX in cell cycle inhibition and decreased β-catenin activity. Finally, treatment of nude mice with CPX significantly decreased tumor xenograft volume. DISCUSSION/SIGNIFICANCE OF IMPACT: CPX demonstrates anti-tumor properties in esophageal cancer cell lines. The current results justify further research into the mechanism of this inhibition. Additionally, given its established safety in humans, CPX is a potential candidate for repositioning as an adjunct treatment for esophageal cancer.

OBJECTIVES/SPECIFIC AIMS: Our main objectives are to study sensory encoding in the adult cortex and quantify rodents’ ability to use intracortical microstimulation to guide behavior. METHODS/STUDY POPULATION: Three rats were implanted with unilateral bipolar stimulating electrodes. The electrodes were connected to a wireless neural stimulator housed in the rat’s backpack. The rat’s swim path was tracked by a video camera above the circular pool, and stimulation parameters were updated in real-time based on distance from the platform. Stimulation was delivered as the distance from the platform increased. Stimulation amplitude was determined through behavioral threshold testing, and parameters ranged from 15–75 μA with 100-Hz pulse trains and 0.2-ms pulses. Rats were first challenged with the 4-platform task in which the submerged platform was randomized across 4 possible locations. This dissociated visual cues from the platform location, as rats had knowledge of the 4 possible locations, but had to use stimulation to guide them efficiently. Next, rats were tasked with the more challenging random-platform task. Visual cues were completely dissociated from the platform location by randomizing the platform location across the entire pool. Performance using the neuroprosthetic device was assessed by comparing trials when the device was on (stimulation trial) Versus off (no-stim trial) for the 2 tasks. RESULTS/ANTICIPATED RESULTS: 4-platform task: Rats visited less potential platform locations when the neuroprosthetic was on Versus off. Rats were also more likely to visit the correct platform location on their first swim trajectory when brain stimulation was delivered. When artificial cues were not available, rats had a greater chance of visiting the platform location from the previous trial. This indicated that rats relied on visuospatial memory without the neuroprosthetic. Random platform task: Performance was measured by taking the ratio of the rat’s actual path length to the optimal path length. When the neuroprosthetic was on, rats demonstrated superior performance through a smaller path to length ratio compared with when the device was off. The platform locations of catch trials were matched to a random subset of stimulation trials, permitting a paired sample t-test. Both rats had significantly shorter path lengths when the device was on. DISCUSSION/SIGNIFICANCE OF IMPACT: Rodents have excellent navigation skills that have been well studied. They have been shown to rely on multimodal sensory information from visual, olfactory, auditory, and idiothetic cues to navigate through their environment. The importance of these cues depends on both their environmental presence and task relevance. In the original Morris water maze experiment, rats use vision to form a visuospatial map of the platform location for allocentric navigation. Here, we have shown that sensory augmented rats can pick up on novel sensory information delivered through ICMS to efficiently find a hidden platform when visual cues are made irrelevant.Our results have implications for the design of the bi-directional sensorimotor neuroprosthetic. We have demonstrated that mammals can interpret artificial sensory information to guide behavior. Future directions include investigating sensory encoding in other primary sensory areas and downstream targets along the somatosensory neuraxis.

OBJECTIVES/SPECIFIC AIMS: This study will assess the effect of essential amino acid (EAA) supplementation on plasma triglyceride (TG) in elderly adults. We will also explore the mechanisms mediating EAA mediated changes in fat metabolism and to suggest promising routes to refine therapy of hypertriglyceridemia. METHODS/STUDY POPULATION: In total, 7 nondiabetic male and female subjects ages 50–75 years with elevated plasma TG levels (130–500 mg/dL) were recruited to participate in an acute (5 h) and long-term (8 wk) EAA supplementation study. We measured changes in regional and whole body fat metabolism, including changes in body composition, plasma TG levels, whole body fat metabolic rates, tissue mitochondrial respiratory capacity, and metabolomic profiles before and after supplementation. RESULTS/ANTICIPATED RESULTS: Long-term EAA supplementation decreased fasted plasma TG levels by 19% (p<0.01). Metabolomics of skeletal muscle found acute EAA supplementation resulted in increased EAA metabolic products while long-term supplementation resulted in increased anaplerosis [flux into the tricarboxylic acid cycle (TCA) intermediate pool] and anaplerotic substrates [propionyl (p<0.01) and succinyl (p<0.01) carnitine] and intermediates of long-chain fatty acid metabolism [stearoyl (p<0.01) and myristoyl (p<0.05) carnitine]. However, tissue level respiratory capacity appeared to be unaffected by EAA supplementation. DISCUSSION/SIGNIFICANCE OF IMPACT: EAA supplementation has potential to improve lipid metabolism and plasma TG levels in non-diabetic older adults. Mitochondrial metabolomics suggest that insufficient TCA pool size may limit tissue fatty acid oxidation and may provide an additional route for nutritional therapy.

Objectives/Goals: Kentucky (KY) is a high priority ending the HIV epidemic state, with high rates of new HIV diagnoses tied to injection drug use. The overall goal of this pilot is to launch sentinel surveillance of bloodborne infections and drug compounds among people who inject drugs (PWID) to monitor trends in near-real time and inform rapid community response. Methods/Study Population: In collaboration with the Clark County, KY, syringe service program (SSP), the pilot study involves two 1-month waves of data collection: enrolling eligible SSP participants and conducting anonymous behavioral surveys, collection of participants’ syringes, laboratory testing of syringes to detect HIV and hepatitis C (HCV), drug residue testing through National Institute of Standards and Technology, and statistical modeling approaches to produce outputs of bloodborne infection and drug detection. Syringes are tested from each enrolled individual for: 1) HIV antibody; 2) HCV antibody; 3) HIV and HCV PCR; 4) HIV antigen; and 5) drug residue. Collaboration with community and PWID stakeholders will identify optimal messaging for reporting results. Results/Anticipated Results: The first wave community-facing pilot was conducted in September–October 2024. 29 survey responses were obtained; median age of the sample is 42 years, 55.2% are gender female; 37.9% reported unstable housing in the past week. Primary drugs of injection reported via survey in the prior month were methamphetamine (62.1%), heroin (13.8%), fentanyl (13.8%), buprenorphine (10.3%), meth and fentanyl in combination (3.4%). PWID reported returning 900 used syringes and a median of 15 per participant visit. At most recent testing, 69.0% reported a positive HCV test; 0% reported a positive HIV test. Some level of drug checking with fentanyl test strips in past month was reported by 51.7%. Initially, 20 syringes were tested for drug compounds; results are pending. HIV and HCV detection testing will be completed by early 2025. Discussion/Significance of Impact: Early results document proof of concept for our sentinel surveillance study; all individuals screened were willing to participate in surveys and syringe collection. New methods to identify risk for disease outbreaks and emerging drugs can inform rapid allocation of prevention resources at a community level, especially where testing is infrequent.

OBJECTIVES/GOALS: The goal was to create and deploy an intuitive, easy-to-use tool that clinical investigators can apply to their data to identify erroneous or inconsistent data entries. Investigators can then correct any errors prior to sharing the data with their statistician for analysis. METHODS/STUDY POPULATION: We developed an interactive shiny app, the Data Loofah, using R Studio that researchers or data analysts can use to examine data. After an investigator uploads data, the app reports which variables are numeric or categorical. Means, standard deviation, median, 25th and 75th quantiles, range and number of missing values are reported for numeric variables. Counts and percentages of categorical variables are summarized. Graphical displays further enhance identification of errors. Access to the Data Loofah is through a secure, university-maintained website with access restricted to university personnel. Supporting materials consisting of instructional step-by-step handouts and videos were developed to assist investigators in the use of the app. RESULTS/ANTICIPATED RESULTS: We will integrate use of the Data Loofah into our Clinical and Translational Science Program’s biostatistics consultative practice. Investigators will use the Data Loofah to pre-screen their data prior to sending it to a statistician, identify errors and inconsistencies, and facilitate making necessary corrections. Statisticians will also use the Data Loofah to review data with investigators prior to starting analyses. Through use of this app, investigators are expected to develop a better understanding of their data specifically and more generally about requirements for preparing data for statistical analysis. Most significantly, regular use of the Data Loofah is expected to result in higher quality data and more efficient use of statistician resources due to reduced effort for data cleaning. DISCUSSION/SIGNIFICANCE: Data cleaning is a time-consuming task and finding data errors can be difficult for data analysts not familiar with clinical variables under study. Further, failure to identify data errors can lead to erroneous results. By facilitating identification of data errors by clinical investigators, the Data Loofah will improve and enhance research output.

OBJECTIVES/SPECIFIC AIMS: Accumulation of cholesterol-laden macrophages in arterial walls leads to atherosclerosis. LXRs induce expression of genes that are atheroprotective in macrophages including CCR7, a chemokine receptor that promotes their emigration from the plaque. CCR7 expression has been shown to be negatively regulated by phosphorylation of LXRα at S198 and is reduced in diabetic mice that show impaired plaque regression. I hypothesized that LXRα phosphorylation at S198 diminishes macrophage emigration from atherosclerotic plaque and contributes to impaired regression in diabetes. METHODS/STUDY POPULATION: Inducible LXRα S198A phosphorylation deficient knock in mouse were used as donors for bone marrow transplantation into mice prone to develop atherosclerosis. Plaques were developed by placing mice on western diet; and regression was induced by lowering their lipid levels. Aortic plaques were then analyzed by using morphometric, histological, and molecular analyses in control and diabetic mice expressing either LXRα WT or LXRα S198A during regression. RESULTS/ANTICIPATED RESULTS: Surprisingly, lack of phosphorylation increased plaque macrophage content and impaired regression under normoglycemic condition; however, it did not exacerbate diabetic regression. Plaques in diabetic mice were associated with increased LXRα S198 phosphorylation. Consistent with this, LXRα phosphorylation is enhanced in macrophages cultured under hyperglycemic conditions indicating glucose-dependent regulation of LXRα phosphorylation. Monocyte trafficking studies reveal that lack of phosphorylation and diabetes independently increase recruitment of monocytes in the plaque that might contribute to increased macrophage content. Importantly, I found that diabetes also increases macrophage retention in the plaque, which is reversed in the absence of phosphorylation. We predict that this increased retention results from inhibition of emigration of plaque macrophages through enhanced phosphorylation in diabetes. DISCUSSION/SIGNIFICANCE OF IMPACT: These findings suggest that inhibiting LXRα phosphorylation could be beneficial in diabetic atherosclerosis to reverse the accumulation of macrophages in the plaque. This study imparts insight on regulation of plaque macrophage trafficking through LXRα S198 phosphorylation.

Objectives/Goals: Recruiting and enrolling participants in health research can be challenging for both the public and study teams. The University of Minnesota (UMN) developed a mobile-friendly, public-facing tool to increase research awareness and connect individuals with active studies that match their interests. Methods/Study Population: UMN developed StudyFinder, which integrates with the clinical trial management, electronic IRB, and ClinicalTrials.gov systems to provide accurate, current study listings with plain-language summaries. The tool pages include advanced Elasticsearch with filters, direct email links to study teams, nightly data refreshes, and an admin dashboard for easy content management and homepage updates. Results/Anticipated Results: Since launch, StudyFinder has received over 1.4 million page views. From Sept 2024–Aug 2025, the site had 78,000 visits, with 10,782 contact emails sent from potential participants to study teams. Analytics show continued year-over-year growth in traffic and engagement, demonstrating StudyFinder’s effectiveness in connecting the public to active research opportunities. Discussion/Significance of Impact: StudyFinder offers a scalable, shareable model for improving public access to research participation. Its open-source code and no-cost license enable other institutions to adopt and adapt the platform, advancing a national ecosystem web-based recruitment. StudyFinder is now available at eight hubs.

OBJECTIVES/SPECIFIC AIMS: The objectives of this study were to compare different methods for determining the disciplines involved in a research article. We sought to address the following questions: To what extent does the number of disciplines reported by an article’s corresponding author agree with their description of the article as unidisciplinary or interdisciplinary? (Q1) and To what extent does the corresponding author’s description of the research as unidisciplinary or interdisciplinary agree with its classification as unidisciplinary or interdisciplinary based on the affiliation of its co-authors? (Q2). METHODS/STUDY POPULATION: Using Scopus, we randomly selected 100 articles from 2010 and 2015 from science teams that had at least 1 author affiliated with Johns Hopkins. Author affiliations were grouped into common academic disciplines: Basic Science, Medicine (and all clinical specialties), Public Health, Engineering, Social Science, Computer Science, Pharmacy, Nursing, and Other. Articles with more than 1 discipline were considered, interdisciplinary. We then sent an online Qualtrics survey to the corresponding author of each article and asked them to indicate (1) all of the disciplines that contributed to the research article at hand, and (2) to indicate whether they considered the research to be “unidisciplinary” or “interdisciplinary” based on definitions that we provided. RESULTS/ANTICIPATED RESULTS: For Q1, we asked corresponding authors to indicate the number of disciplines involved in their research and then to choose the definition that best described their research. Among 76 respondents, 42 indicated that their research consisted of 1 discipline, and 34 indicated that their research consisted of more than 1 discipline. Of the 42 respondents who indicated that their research consisted of one discipline, 21 (50%) respondents described their research as “unidisciplinary” and 21 (50%) described their research as “interdisciplinary.” However, of the 34 respondents who indicated that their research consisted of more than 1 discipline, all but 1 (97%) described their research as “interdisciplinary.” For Q2, we assigned a discipline to each co-author based on his/her affiliation and counted the number of disciplines involved. Among 76 respondents, of the 22 who described their research as “unidisciplinary,” 16 (73%) were categorized as “unidisciplinary” and 6 (27%) were categorized as “interdisciplinary,” using this method. Of the 54 respondents who described their research as “interdisciplinary,” 30 (56%) were categorized as “interdisciplinary” and 24 (44%) as “unidisciplinary.” DISCUSSION/SIGNIFICANCE OF IMPACT: Our results highlight that different methods for determining whether a given research article is interdisciplinary are likely to yield different results. Even when researchers indicate that their research is based within one major discipline, they may still consider it interdisciplinary. Likewise, classifying an article as either unidisciplinary or interdisciplinary based on the affiliations of its co-authors, may not be consistent with the way it is viewed by its authors. It is important to acknowledge that assessing the interdisciplinarity of research is complex and that objective and subjective views may differ.