OBJECTIVES/GOALS: This project focuses on investigating the potential of Simalikalactone D (SKD) as an anticancer agent, exploring the mechanisms underlying SKD’s induction of cell death, and assessing the impact of SKD on diverse breast cancer cell lines. Also, it Investigates the compound’s mechanisms of action beyond caspase 3-dependent pathways. METHODS/STUDY POPULATION: Three breast cancer cell lines were used: SKBR3, MDA-MB-231, and MDA-MB-468. Two triple-negative breast cancer cell lines are included to address cancer disparities across diverse ethnic backgrounds. Viability assays were conducted to determine half-maximal inhibitory concentrations (IC50). Caspase 3 activity assay was performed to evaluate apoptosis as a possible cell death pathway. Wound healing and colony formation assays are used to assess cell migration and clonogenic capacity. Proteomic analysis and phosphoarray analysis are planned for a deeper understanding of SKD’s anticancer properties, as well as testing for caspase 3 independent pathways. RESULTS/ANTICIPATED RESULTS: SKD demonstrated substantial cytotoxicity against all three breast cancer cell lines. IC50 values for SKBR3, MDA-MB-231, and MDA-MB-468 were 60.0 nM, 65.0 nM, and 116 nM, respectively. SKD induces cell death via caspase 3-independent pathways. Further experiments are needed to confirm and elucidate the molecular pathways being impacted. SKD inhibited cancer cell migration and clonogenic potential, suggesting it can reduce tumor growth and metastatic tendencies. DISCUSSION/SIGNIFICANCE: The study highlights SKD’s cytotoxicity across diverse breast cancer cell lines. It underscores the mechanism of action, a caspase 3 independent pathway. These findings hold promise for the development of innovative anticancer treatments and emphasize the importance of exploring varied cellular responses to mitigate global cancer disparities.

OBJECTIVES/GOALS: BURRITO is an efficient strategy that provides full disclosure in the electronic medical record of a patient’s preference in real time. BURRITO uses printed materials only to inform patients and has a <50% rates of consent. We hypothesized that adding an informational video to the printed materials would increase donations. METHODS/STUDY POPULATION: This study was IRB-approved and was considered minimal risk. The BURRITO self-consent workflow process (Soares et. al, Biopreservation and Biobanking, IN PRINT) was developed in an outpatient cardiology clinic. In the same clinic, patients were randomized to receiving printed materials only (standard procedure) or the printed materials plus a 2.5-minute informational video (intervention) while waiting for the physician in the exam room. Randomization occurred at the level of the day in clinic. Patients were blinded to the nature of the study. Following the presentation of information, the patient’s decision on consent for donation was documented in the electronic record by ancillary clinical staff. Rates of consent were analyzed by a statistician not involved in the experiment and after completion of trial. RESULTS/ANTICIPATED RESULTS: Thirty-five clinic days were randomized to either intervention (17 days) or standard (18 days), and a total of 255 patients decided during their visit to either “opt-in” or “opt-out” to donating remnant biospecimens for future research. One hundred patients opted to defer deciding (28%). No significant demographic differences were noted between the study arms. The rate of consent was 73% vs. 58% in the intervention group and the control group, respectively (p-value = 0.014). This represents an increase in the odds of consenting with an informational video by 96% (OR = 1.96, 95% CI = 1.15 to 3.34). DISCUSSION/SIGNIFICANCE OF IMPACT: This is the first randomized trial to show that an informational video with printed materials is superior for when patients are self-consenting to opt-in for clinical remnant biospecimen donation. This result adds to the evidence that the BURRITO process plus video (BURRITOv) is an effective approach for biospecimen universal consenting.

OBJECTIVES/GOALS: Test the validity of a system for subgrouping people with CLBP by comparing lumbar spine alignment in two CLBP subgroups and sexes during clinical tests of maximum flexed and extended sitting and a functional test of preferred sitting. METHODS/STUDY POPULATION: Using the Movement System Impairment classification system, 154 participants with CLBP were subgrouped based on the predominant direction of altered movement and alignment patterns and symptoms during a standardized examination. Participants performed a functional test of preferred sitting followed by clinical tests of maximum flexed and extended sitting in random order. Reflective markers were place superficial to T12, L3 and S1 spinous processes. 3D marker co-ordinate data were collected using an 8 camera motion capture system. Sagittal plane lumbar curvature angle (LCA), defined as the angular distance between T12, L3, and S1 landmarks was calculated for each test. A three-way mixed effect ANOVA model was used to examine the following effects: test, subgroup, sex, test*subgroup, test*sex, subgroup*sex. RESULTS/ANTICIPATED RESULTS: Test: Lumbar alignment patterns were different for flexed [LCA = 7.4° (6.1, 8.7)], extended [LCA = −22.6° (−23.9,−21.3)], and preferred [LCA = −3.8° (−5.2,−2.5)] sitting tests. LBP subgroup: Rotation-extension [LCA = −7.5° (−8.7,−6.3)] had more extended lumbar alignment than rotation [LCA = −5.2° (−6.2,−4.2)]. Sex: Women had more extended lumbar alignment [LCA = −10.3° (−11.2,−9.3)] than men [LCA = −2.5° (−3.7,−1.2)]. Test*sex: The difference in lumbar alignment between women and men was smaller during the flexed sitting test [women = 4.2° (2.5, 5.9), men = 9.9° (7.8, 12.1)], compared to extended [women = −27.5° (−29.2, −25.8), men = −17.0° (−9.2, −14.8)] and preferred [women = −7.4° (−9.1, −5.8), men = −0.3° (−2.5, 1.8)]. The test*subgroup (p = 0.84) and subgroup*sex (p = 0.87) interactions were not significant.

Objectives/Goals: Longitudinal studies capture dynamic host–environment interactions crucial to microbiome research. Commonly used mixed-effects models can struggle with small clinical samples, like many endometrial cancer studies. We will compare them to simpler fixed-effects models, aiming to develop a new fixed-effects differential abundance alternative. Methods/Study Population: Starting with simulated data for linear models with a single treatment effect, we will incrementally increase complexity to reflect real-world microbiome data. To observe conditions of misspecification, simulated data of repeated measures will be generated using different distributions, while varying parameters and inter-class correlation. In each condition, linear fixed- and mixed-effects models will be fitted 1000 times to empirically determine p-value, type 1 error rate, and power. We will increase the verisimilitude of our approach by using endometrial cancer data from our lab as a real background, while adding simulated signals. This data will be used to assess extant differential abundance models against our novel fixed-effects model. Results/Anticipated Results: Our initial findings indicate that fixed-effects models maintain control over type 1 error rates in small samples, while mixed-effects models do not perform as reliably. We are now exploring the effects of confounders on type 1 error rates and power in linear models, while our next step will evaluate generalized linear models in a differential abundance context. We expect that the fixed-effects models will continue to be as reliable as mixed-effects models, while being less computationally complex. After establishing our theoretical basis, our novel model should perform comparably to mixed-effects models on pseudo-simulated endometrial cancer data and better at small sample sizes. We will finally apply our model to real endometrial cancer data to identify microbial markers of disease predisposition. Discussion/Significance of Impact: By characterizing and improving these statistical tools, we seek to increase the robustness and power of microbiome study results (especially at small sample sizes), ultimately supporting better-informed translational research decisions. This work underscores the importance of statistical methods in advancing microbiome research.

Objectives/Goals: To develop and validate a Random Survival Forest model predicting individualized tactile sensory recovery after DIEP flap breast reconstruction, enabling personalized postoperative counseling and future clinical decision-support integration. Methods/Study Population: Neurotized DIEP flap reconstructions (2019-2025) with pressure-specified sensory device testing were analyzed for time to ≥ 50% tactile sensory recovery. A five-variable Random Survival Forest (RSF) (dual vs. single coaptation, BMI, age, operative time, mastectomy weight) was trained on an imputed dataset. Discrimination (C-index; 1 = perfect discrimination, 0.5 = random chance), 12-month accuracy (Brier score; 0 = perfect forecast, 1 = completely inaccurate forecast), and calibration (predicted vs observed recovery quartiles) were assessed. Representative cases illustrate patient-specific recovery curves. Results/Anticipated Results: Of 110 breasts, 60 achieved ≥ 50% recovery. Mean follow-up was 17.1 ± 1.7 months (median 12.8, 95% CI 9.7-19.7). The RSF achieved C-index 0.69. The mean predicted recovery increased from 32.3% at 6.13 months to 49.5% at 11.8 months, 56.2% at 17.7 months, and 60.8% at 24.1 months. Calibration at 12 months by quartiles of predicted recovery showed observed recovery 0% (Q1), 48.8% (Q2), 80.8% (Q3), and 100% (Q4). The corresponding mean predicted probabilities were 17.2%, 43.1%, 62.3%, and 77.0%, respectively. The RSF model demonstrated superior predictive accuracy (Brier = 0.099) compared with a Kaplan-Meier baseline (Brier = 0.247). The low-recovery case had predicted recovery 6.2% at 12 months. The median case tracked the cohort mean: 52.4%. The high-recovery case achieved 81.8% at 12 months. Discussion/Significance of Impact: A five-variable RSF provides individualized recovery trajectories with moderate discrimination and good internal calibration. The tool quantifies time-dependent probabilities and supports the development of a perioperative counseling tool to predict expected tactile recovery after neurotized DIEP flap breast reconstruction.

OBJECTIVES/SPECIFIC AIMS: To study the biology of Phosholamban (PLN) in a human relevant model. METHODS/STUDY POPULATION: State of the art stem-cell technologies using iPSC-CMs derived from carriers of a lethal PLN mutation. RESULTS/ANTICIPATED RESULTS: Our preliminary data demonstrate that this particular PLN mutation (L39) results in reduced expression and mis-localization of PLN as well as increased incidence of early after depolarization in isolated iPSC-CMs. DISCUSSION/SIGNIFICANCE OF IMPACT: Phospholamban (PLN) is a critical regulator of Ca++ homeostasis yet many uncertainties still remain regarding its role in humans. Our study will provide unique insights into the pathophysiology of this protein in HF.

ABSTRACT IMPACT: Given the association between lower time to treatment and better clinical outcomes in stroke patients, identifying factors correlated with reduced proximity and thus greater time to stroke care can aid efforts to reduce disparities in stroke outcomes. OBJECTIVES/GOALS: The objective of this study is to quantify the relationship between distance to the nearest certified stroke hospital and census-derived demographics of age, race/ethnicity, income, and insurance status. METHODS/STUDY POPULATION: This is a cross-sectional study. Population data for all census tracts in the contiguous United States were obtained from the US Census Bureau’s 2014-2018 American Community Survey. Stroke hospitals were identified from national or state level certification databases and were required to offer at least IV tPA. The main outcome is driving distance in kilometers from each census tract to the nearest certified stroke center, which was calculated using OSMnx, a Python package to retrieve, model and analyze real-world street networks. Quantile regression analysis was used to compare relationships between distances and tract-level demographics of age, race/ethnicity, income, and insurance status. RESULTS/ANTICIPATED RESULTS: 2,423 stroke centers and 71,929 census tracts containing 316,995,649 individuals were included. 49,918 (69%) tracts were urban. Demographic disparities in proximity to certified stroke care were greater in non-urban areas than in urban areas. Higher representation of individuals with age ≤65 years were associated with increased median distance to a certified stroke center in non-urban areas, but not urban areas. Median distance was greater with greater representation of American Indian or uninsured populations in urban and non-urban census tracts. Higher median income was associated with decreased median distance in non-urban census tracts and greater median distance in urban census tracts. DISCUSSION/SIGNIFICANCE OF FINDINGS: Reduced proximity to stroke care exists in areas with greater representation of elderly, American Indian, or uninsured persons; and low median income. These disparities are magnified in non-urban settings. Such knowledge can aid efforts to address and reduce disparities in stroke outcomes.

OBJECTIVES/GOALS: To increase diversity in clinical and translational research (CTR), to strengthen protections for uninsured and under-insured study participants from vulnerable groups by addressing their medical, ancillary care and psycho-social needs and to develop a systematic ethically sound approach to addressing such needs in the study protocol and budget. METHODS/STUDY POPULATION: We conducted) ethical analyses of: (1) the regulatory and ethics scholarship concerning protections and duty of care to research participants from vulnerable groups, and 2) arguments concerning the nature and scope of ancillary care obligations of researchers, as well as 3) a review of the applicable local, federal, and international regulations and practices concerning the duty of care to CTR participants and potential participants who are uninsured, under-insured and/or undocumented members of vulnerable groups. RESULTS/ANTICIPATED RESULTS: Uninsured and underinsured study participants pose major ethical challenges for CTR as medical needs arising during a study are usually covered by the participants own insurance. Lack of health insurance increases vulnerability and creates (1) a barrier to research participation for members of socially disenfranchised groups, (2) risk of discriminatory exclusion of such participants from clinical studies, and (3) inter-institutional inconsistencies in meeting their medical needs; thus limiting diversity in CTR. To address the challenges, we propose an inclusive, systematic, ethically sound approach, which deliberately plans for and provides resources within a study protocol to address the medical and psycho-social needs of uninsured participants during and beyond the study. DISCUSSION/SIGNIFICANCE: Including diverse participants in CTR ensures data quality and social justice. PIs and IRBs should adopt an inclusive approach to the medical needs of vulnerable uninsured participants, plan for their medical and ancillary care needs in the protocol and budget, list community resources, provide follow-up support and note assistance in their files.

Objectives/Goals: The primary objective of this research is to examine current implementations of Artificial Intelligence (AI) models in clinical trials, focusing on advantages, limitations, and ethical considerations associated with their application in pediatric populations. Methods/Study Population: A comprehensive literature search was conducted using PubMed and Google Scholar databases. The initial search used the key terms “artificial intelligence” and “pediatric.” The results were refined to include publications from 2023 to 2025. The keyword “imaging” was added, focusing on studies exploring AI-based diagnostic tools. Articles were included if they had at least one of the following: (1) the need for pediatric-specific databases, (2) diagnostic accuracy within pediatric populations, or (3) ethical considerations in AI-driven pediatric research. Data were retrieved from ClinicalTrials.gov, where “artificial intelligence” was specified. Eligibility criteria were limited to pediatric participants and restricted to studies conducted in the USA to ensure regulatory consistency. Results/Anticipated Results: An analysis of trials from ClinicalTrials.gov further highlighted the underrepresentation of pediatric subjects in AI-related clinical research, emphasizing a critical gap in age-appropriate AI model development and validation. AI models demonstrate great potential for improving diagnostic accuracy, but effectiveness remains limited by lack of pediatric-specific training data. In pediatric chest imaging, AI algorithms achieved diagnostic performance comparable to adult populations in detecting bacterial pneumonia (Morcos et al., 2023). Conversely, a large-scale review of public medical imaging datasets revealed an increase in false-positive rates among younger patients, reflecting model performance disparities (Zamora et al., 2024). Discussion/Significance of Impact: Implementing AI-driven models can accelerate data analysis and streamline trial design. The application of AI models in pediatric clinical trials can accelerate the timeline for treatment and reduce misdiagnosis. Addressing data gaps and ethical considerations about consent and algorithmic bias are crucial to improve pediatric health outcomes.

OBJECTIVES/SPECIFIC AIMS: Deficits in reward-based learning have been shown in youth at risk for developing substance use disorders (SUD). Here, we investigated whether computational models can be used to more precisely delineate the additive effects of such risk loading (i.e., the comparison between youth with ADHD, and those with ADHD and familial SUD) on reward-based learning in youth. METHODS/STUDY POPULATION: In total, 41 drug-naïve youth, stratified into 3 groups based on ADHD diagnosis and parental SUD: healthy controls (HC, n=13; neither ADHD nor parental SUD), low risk (LR, n=13; ADHD only), and high risk (HR, n=15; both ADHD and parental SUD), performed a reward task. Learning rates, prediction and congruence t-scores were computed using a reinforcement learning model and analyzed via a multivariate ANOVA. RESULTS/ANTICIPATED RESULTS: The analyses showed a significant linear effect in task accuracy, which decreased with increasing risk profiles. Analyses of the model-derived variables also showed similar significant linear effects in learning rates and the congruence t-score, but not in the prediction t-score. These effects were primarily driven by significantly higher learning rate and congruence t-score in HC compared with HR youth. DISCUSSION/SIGNIFICANCE OF IMPACT: These results show most profound deficits in reward-learning in HR youth. These findings also show that computational analyses can offer added value over conventional behavioral analyses by more precisely evaluating group differences in relation to SUD risk.

ABSTRACT IMPACT: Our work helps show universities that embedding dedicated sexual health clinics within university health and wellness clinics may expand the amount of students they see for sexual health screenings during a time of increased sexual behavior and exploration. OBJECTIVES/GOALS: The National College Health Association reports that college students have frequent, condomless sex. Student health and wellness clinics (SHWC) offer sexual health services, but few have dedicated sexual health clinics (SHC). We evaluated sexual health service use at a university SHWC after implementation of a dedicated SHC two half-days per week. METHODS/STUDY POPULATION: This was a retrospective analysis of data collected from patients receiving sexual health screening at the University of Alabama at Birmingham (UAB) SHWC between January 2015 and June 2019. Demographic variables, sexual behaviors, reason for testing, and rates of STIs were extracted from the electronic medical record and were compared by clinic (SHC vs. SHWC). Data on screening visits of patients over 18 were included in the final analysis. Variables were summarized with frequencies and percentages. Univariate models were fit, and multi-variable models will be fit, selecting variables with p values of 0.1 or less. Odds ratios with corresponding 95% confidence intervals for univariate analysis are presented. The study was approved by the UAB Institutional Review Board. RESULTS/ANTICIPATED RESULTS: A total of 5025 STI screenings were performed. Males (OR 4.13; 3.61-4.72), undergraduates (OR 1.33; 1.15-1.54), and persons reporting sex with the same sex (OR 1.88; 1.56-2.28), were significantly more likely to seek care at the SHC. Students with symptoms were more likely to seek care at the SHWC (OR 0.53; 0.47-0.61), while persons who reported contact with STIs were more likely to seek care at the SHC (OR 2.88; 2.22-3.74). The overall percentage of positive screenings was 9.3% for chlamydia (CT), 3.0% for gonorrhea (GC), 0.8% for trichomoniasis (TV), 0.7% for syphilis, and 0.3% for HIV with higher percentages of positive for CT (OR 1.60; 1.30-1.96) and GC (OR 2.02; 1.44-2.85) in the SHC. A greater percentage of positives for TV (OR 0.37; 0.14-0.96) was found in the SHWC. DISCUSSION/SIGNIFICANCE OF FINDINGS: Based on demographics of persons utilizing services, embedding a dedicated SHC within a university SHWC may expand populations reached for STI screening. With higher percentages of patients testing positive for CT and GC, a SHC may allow for greater diagnosis and treatment of STIs in general screening and persons presenting as contacts.

OBJECTIVES/GOALS: To evaluate the incidence of brachial plexus birth injury (BPBI) and its associations with maternal demographic factors. Additionally, we sought to determine whether longitudinal changes in BPBI incidence differed by maternal demographics. METHODS/STUDY POPULATION: We conducted a retrospective cohort study of over 8 million maternal-infant pairs using California’s Office of Statewide Health Planning and Development Linked Birth Files from 1991-2012. Descriptive statistics were used to determine BPBI incidence and the prevalence of maternal demographic factors (race, ethnicity, age). Multivariable logistic regression was used to determine associations of year, maternal race, ethnicity, and age with BPBI. Excess population level risk associated with these characteristics was determined by calculating population attributable fractions. RESULTS/ANTICIPATED RESULTS: The incidence of BPBI between 1991-2012 was 1.28 per 1000 live births, with peak incidence of 1.84 per 1000 in 1998 and low of 0.9 per 1000 in 2008. Incidence varied by demographic group, with infants of Black (1.78 per 1000) and Hispanic (1.34 per 1000) mothers having the highest incidences. Controlling for relevant covariates, infants of Black (AOR=1.88, 95% CI 1.70, 2.08), Hispanic (AOR=1.25, 95% CI 1.18, 1.32) and advanced-age mothers (AOR=1.16, 95% CI 1.09, 1.25) were at increased risk. Disparities in risk experienced by Black, Hispanic, and advanced-age mothers contributed to a 5%, 10%, and 2% excess risk at the population level, respectively. Longitudinal trends in incidence did not vary among demographic groups. Population-level changes in maternal demographics did not explain changes in incidence over time. DISCUSSION/SIGNIFICANCE: Although BPBI incidence has decreased in California, demographic disparities exist. Infants of Black, Hispanic, and advanced-age mothers are at increased BPBI risk compared to White, Non-Hispanic, and younger mothers.

Objectives/Goals: This study demonstrates the utility of the CBID biodesign process for identifying and prioritizing high-impact neurosurgical needs. The research emphasizes the process’s role in developing innovative medical technologies that align with the healthcare ecosystem’s demands and stakeholder priorities. Methods/Study Population: The CBID Spiral Innovation Model, integrating clinical, technical, business, and strategic considerations across clinical challenges in neurosurgery was employed over a 15-week period at a tertiary care center. The process involved three phases: (1) needs identification through 8 weeks of clinical immersion, (2) 7–8 weeks of stakeholder engagement via informational interviews, surveys, and conferences, and (3) iterative refinement based on evidence generation and market value. Stakeholders included over 70 clinicians (neurosurgeons, neurocritical care specialists, neurologists, etc.) across 15 institutions as well as more than 10 payers and hospital administrators. Data collection encompassed direct observation, structured interviews, and comprehensive literature review. Results/Anticipated Results: The initial list of 300+ identified neurosurgical needs was reduced to 271 after clinician and market input. High-level market and clinical evidence assessments further reduced this to 74 needs. Finally, through iterative evaluation of evidence generation, market opportunity, and stakeholder feedback, five critical unmet needs in stroke, traumatic brain injury, hydrocephalus, and epilepsy were identified for technological innovation. These needs met the criteria for clinical importance, economic viability, and market accessibility. The findings highlight the effectiveness of the biodesign process in creating a roadmap for innovation that is both clinically relevant and commercially viable. Discussion/Significance of Impact: This study underscores the effectiveness of structured need-finding and prioritization within neurosurgery. Integrating stakeholder perspectives and rigorous analysis, it provides a replicable framework for medical innovation to accelerate the development of impactful solutions across medicine.

OBJECTIVES/GOALS: Biomedical research fields are facing the challenges of demand for skilled workers as well as challenges related to diversity in that workforce. It is important that the healthcare workforce reflect the population it serves. The Exposures Internship seeks to address this by building pathways for youth to pursue careers in research and medicine. METHODS/STUDY POPULATION: In 2021, the Yale Cultural Ambassadors expressed concern about the lack of free high quality, educational offerings for youth that summer. They asked YCCI to consider developing a summer program for students aged 15 and older that focused on spurring interest in careers in healthcare, medicine, and clinical and translational research. The result was a 4-week virtual learning experience for 34 interns who met daily via Zoom and participated in course work, lectures, journal clubs, group projects, and virtual lunches with internationally renowned clinical research and healthcare leaders. Sessions were designed to help interns gain knowledge of and exposure to current topics in clinical and translational science and to observe the various steps of proposing, designing, undertaking, and analyzing clinical trials. RESULTS/ANTICIPATED RESULTS: YCCI received over 900 inquiries from around the world with more than 200 completed applications for participation in the internship for the pilot year. Since then, YCCI leadership has worked with community partners to engage young scholars from 17 different states, Canada, Mexico and Puerto Rico. Of those, we estimate 75% are minority, ~50% female and 20% from rural areas with limited similar opportunities. During the four weeks of the program these highly motivated students worked on projects aimed at increasing participation in pediatric research through a revised Informed consent and adolescent assent process and a youth centered awareness campaign. Interns were so inspired that they requested the program be continued beyond the initial four weeks. As such, YCCI continued to offer sessions throughout the year. DISCUSSION/SIGNIFICANCE: In evaluation of the pilot program 95% of respondents strongly agreed that the program exposed them to new information about clinical and translational research. One intern shared, This program has unquestionably made me consider becoming a researcher in the future with the goal of becoming a principal investigator within my interest in medicine.

OBJECTIVES/SPECIFIC AIMS: Individuals with autism spectrum disorder (ASD) and developmental coordination disorder (DCD) share overlap in their motor symptom profile and underlying neurology (Sumner, Leonard, & Hill, 2016, JADD). DSM-5 guidelines allow these 2 disorders to be independent or co-occurring (APA, 2013), but common clinical practice does not include systematic assessment to determine the presence or absence of co-occurring DCD in children with ASD, or vice versa. Furthermore, in many cases DCD is managed in a nonspecific manner, with schools making accommodations for a child’s motor challenges without formally assigning a diagnosis of DCD. Thus, somewhat subjective, qualitative judgments are made by clinicians to classify children as DCD, ASD, or ASD+DCD in the absence of a reliable, valid, quantitative measure to distinguish between these profiles. As a first step toward developing such a measure, researchers must tease apart the nuanced differences in the motor symptoms of these 2 developmental disorders using methods that are scalable to clinical and educational settings. These methods must also be developed with consideration for logistical variables such as cost, clinical utility of data output, and ease-of-use if they are to be transferrable to physicians, school nurses, and other community health workers outside of academic laboratory settings. To that end, we conducted 2 complementary studies: 1 in the lab and 1 in the community. METHODS/STUDY POPULATION: In the community-based study, we used an affordable, user-friendly, portable balance testing system to assess postural stability during quiet standing (feet shoulder-width apart) with eyes open for 30 seconds. Data were generated from a single force plate in the balance platform. Potential participants were screened for other medical and neurological conditions that might impact their postural stability, and those with significant comorbidities were excluded. We tested 15 children with a reported diagnosis of ASD, 8 children with suspected or diagnosed DCD who were enrolled in a motor intervention program, and 30 typically-developing (TD) children with no significant developmental history reported. The ASD group ranged in age from 7 to 20, the DCD group ranged from 7 to 10, and the TD group ranged from 7 to 19. In the lab-based study, we again obtained force plate data during quiet standing (feet shoulder-width apart) with eyes open for 30 seconds, in our system that also included full-body motion capture, virtual reality, and mobile eye tracking. (Data from these additional sources are not discussed in this disseminaton, as our current focus is on identifying a simple, scalable metric that can be used to distinguish ASD from DCD.) We tested 10 children with a diagnosis of ASD that was confirmed by the research team, 10 children with a diagnosis of DCD that was confirmed by the research team, and 5 TD children with no significant developmental history reported. The ASD group ranged in age from 7 to 18, the DCD group ranged from 8 to 12, and the TD group ranged from 9 to 18. RESULTS/ANTICIPATED RESULTS: Primary outcome measures in both studies were related to Center of Pressure (CoP), including CoP sway, CoP velocity, and amount of sway relative to the base of support. Data analysis from both studies is ongoing, but preliminary trends suggest that CoP metrics may effectively differentiate between ASD, DCD, and TD. During quiet standing, individuals with DCD exhibited the greatest postural sway. Individuals with ASD followed, having greater instability than the TD group. Differences were also evident in the velocity profiles of postural sway. DISCUSSION/SIGNIFICANCE OF IMPACT: Preliminary findings suggest that CoP offers a means of differentiating between typical and atypical development specifically with respect to motor symptoms. This simple, quantifiable measure may prove a sensitive and specific means of systematically assessing co-occurrence of ASD and DCD in clinical and educational settings, leading to more accurate diagnostic classification and tailored intervention. Future directions include conducting analyses that account for participant age and developmental stage with respect to motor skills, determining whether trends hold in a larger sample, and using advanced statistical methods to determine whether CoP variables have predictive validity in discriminating between classifications of ASD, DCD, ASD+DCD, and TD. Eye-movement data were also obtained during these tasks, and may further aid in understanding the factors contributing to atypical postural control. These 2 studies also yielded methodological findings, namely that the portable force platform carries the benefit of high ease-of-use, low cost, and portability, but also has important drawbacks. Specifically, it is not capable of registering accurate CoP data for participants who weigh <40 lbs, and the error variance for the load cells is greater than that of most nonportable, higher-end plates like those embedded in our laboratory’s platform. As technological advances continue to facilitate development of more portable, higher-resolution systems, these drawbacks may be significantly reduced. Future directions include further assessment of portable, affordable solutions for this type of testing to identify whether higher-resolution options are available, whether this added resolution increases classification accuracy, and how ease-of-use is perceived by clinical and community health workers.

OBJECTIVES/GOALS: The Clinical Research Coordinator (CRC) Intern Program was developed to increase knowledge and awareness of CRC’s vital role in conducting clinical studies. Program outcomes are to provide students with marketable skills and knowledge leading to employment within the clinical research ecosystem. METHODS/STUDY POPULATION: The CRC Intern Program is available to college students for health-related academic courses requiring an internship component. Didactic and experiential learning are incorporated into the program with students imbedded within well-established clinical research teams. Activities include attending IRB meetings, recruitment and enrollment, data collection and entry, and regulatory items. Students complete knowledge pre- and post-assessments (Competency Index for Clinical Research Professionals-CIRCP) via REDCap surveys to assess learner knowledge acquisition and program effectiveness. Demographic, program evaluation, and 3-month follow up survey data are analyzed using descriptive statistics. RESULTS/ANTICIPATED RESULTS: Beginning in Spring 2022, the Intern Program has accepted 9 students with 5 completing the program, with 2 of this 5 having been offered employment as CRCs. Preliminary CIRCP assessment data indicates increased CRC knowledge upon Intern Program completion. Demographic data shows that students are mostly female and non-white (43% African American, 29% Hispanic). Additional results from the current cohort will be shared upon program completion. Of note is the development of partnerships with local colleges, including community colleges, to build awareness of the CRC career path and to provide opportunities for CRC exploration resulting in expansion and diversification of the clinical research workforce. DISCUSSION/SIGNIFICANCE: There is an industry wide shortage of CRCs. Our internship program has provided an effective method to expand and diversify the CRC workforce through knowledge acquisition and application building CRC skills and competencies. Lessons learned and future plans for intern expansion will be discussed.

OBJECTIVES/GOALS: To improve early career faculty members’ NIH grant writing skills, Clinical and Translational Science Awards (CTSA) hubs have developed a variety of workshop-style programs. However, few articles have evaluated the impact of grant writing workshops on NIH grant submission and award rates. METHODS/STUDY POPULATION: The K Writing program was developed by the Michigan Institute for Clinical and Health Research (MICHR) at the University of Michigan. Since 2012, 435 scholars have participated in the program. The MICHR K Writing program is a three-part workshop series that prepares scholars by providing them with guidelines to write all sections of a career development grant application. Each session focuses on different sections of the K award proposal. During the workshop sessions, participants break into small groups and exchange drafts of their proposal sections and receive peer critique and feedback from senior faculty facilitators who have experience with NIH study sections. RESULTS/ANTICIPATED RESULTS: Between 2012-2018, 273 scholars participated and 57% were female. Our two primary outcomes of interest are submission rates and success rates (the number of grants awarded divided by the number of applications). We plan to examine the effects of several characteristics, including number of sessions attended, cohort year, and faculty vs. postdoctoral status. We will also examine whether there were differences in submission and success rates between female and male researchers and between underrepresented minority scholars and those who identified as white or Asian. Lastly, we will report submission and success rates for each grant mechanism and compare them to the national averages. DISCUSSION/SIGNIFICANCE: Obtaining external research funding is an important part of a faculty career, especially at its early stages. This research has important implications for the design of similar programs intended to increase submission and success rates for federal grant applications.

Objectives/Goals: Telehealth is a key solution to improving access to healthcare and disease detection, especially in rural areas. Telehealth access relies on the presence of broadband in an area, which has shifted with the growing importance of internet. We aim to assess the distribution of US county-level broadband availability from 2017 to 2020 overall and by rurality. Methods/Study Population: We employed an ecologic study design to examine the distributions of two measures of broadband availability across all US counties from 2017 to 2020. Broadband presence was defined as counties meeting the Federal Communications Commission (FCC) definition of broadband with at least one high-speed internet provider and an average download/upload speed of at least 25/3 megabits per second using 2017–2020 FCC Broadband Deployment Data. Broadband access, or county-year proportion of households with broadband, was defined using 2017–2020 data from the American Community Survey. We used log-binomial and linear regression models and a difference-in-difference analytical approach to estimate the difference in the change in broadband presence and access from 2017 to 2020 between rural and urban counties. Results/Anticipated Results: Overall, broadband presence increased across the study period, where 66.82% of counties had broadband present in 2017 versus 92.57% in 2020. A major jump occurred from 2018 to 2019, where 64.96% of counties had broadband present in 2018 compared to 91.09% in 2019. Broadband access also rose, where the average proportion of households with broadband was 69.43% in 2017 and 78.24% in 2020. The increase in both broadband presence and access from 2017 to 2020 was larger in rural counties compared to urban counties. Specifically, the increase in the probability of having broadband present from 2017 to 2020 was 30% higher in rural compared to urban counties (95% CI: 1.24, 1.38). The increase in the proportion of households with broadband was 1.21% higher from 2017 to 2020 in rural compared to urban counties (95% CI: 0.004, 0.021). Discussion/Significance of Impact: While broadband presence and access both increased from 2017 to 2020, the observed increases were larger in rural compared to urban counties. Improvements in broadband access in the US are continuously needed to increase the use of telehealth and, subsequently, lessen rural/urban disparities in healthcare access and disease detection.

OBJECTIVES/SPECIFIC AIMS: Treatment of acute myeloid leukemia (AML) is challenging, as apoptosis-resistant AML cells often persist within the bone marrow microenvironment despite chemotherapy. The overall goal of our laboratory is to identify and ultimately target the bone marrow factors that protect AML cells. METHODS/STUDY POPULATION: Using cell cultures, we previously reported that SDF-1 (CXCL12), an abundant bone marrow chemokine, induces apoptosis of isolated CXCR4+ AML cells, including freshly isolated bone marrow-derived AML cells from approximately one-third of AML patients. However, co-culture of AML cells with differentiating osteoblasts protected AML cells from apoptosis. RESULTS/ANTICIPATED RESULTS: Histone deacetylase inhibitors (HDACi) abrogated the ability of osteoblasts to protect AML cells and altered expression of matrix mineralization genes including tissue nonspecific alkaline phosphatase (TNAP). A different drug, cyclosporine A (CSA), similarly inhibited osteoblast-mediated protection of AML cells and reduced TNAP expression. Specifically targeting osteoblast TNAP via siRNA was sufficient to prevent osteoblasts from protecting AML cells in co-cultures. In addition, we are targeting TNAP enzymatically. DISCUSSION/SIGNIFICANCE OF IMPACT: Our results indicate that targeting TNAP may be useful in AML treatment to render the bone marrow microenvironment more hostile to leukemic cell survival.

OBJECTIVES/SPECIFIC AIMS: Glaucoma is a leading cause of irreversible blindness worldwide; in the United States alone, over 2.7 million individuals are affected. Various risk factors for glaucoma are known and include age, race/ethnicity, genetics, and ocular measures. Despite numerous studies, molecular and environmental factors that contribute to glaucoma remain elusive. Our objective was to conduct a genome-wide association for glaucoma among black and white HRS respondents, and to determine the feasibility for future analyses examining shared genetic markers between glaucoma and other comorbidities, behaviors, and environmental risk factors. METHODS/STUDY POPULATION: The University of Michigan Health and Retirement Study (HRS) is a longitudinal survey of a representative sample of Americans over the age of 50. Supported by the National Institute on Aging and the Social Security Administration, the HRS is designed to provide reliable data on the decisions, choices, and behaviors of people as they age and respond to changes in public policy, the economy, and health. The study obtains information every two years about income and wealth, health and use of health services, work and retirement, and family connections. Through its unique and in-depth interviews, the HRS provides an invaluable and growing body of multidisciplinary data that researchers can use to address important questions about the challenges and opportunities of aging. Because of its innovation and importance, the HRS has become the model and hub for a growing network of harmonized longitudinal aging studies around the world. Saliva was collected on half of the HRS sample each wave starting in 2006 and respondents were genotyped on the Illumina Human Omni2.5-Quad (Omni2.5) BeadChip at the NIH Center for Inherited Disease Research. We accessed survey results to evaluate prevalence of glaucoma in this dataset and performed a genome-wide association study (GWAS) adjusting for age, sex, and significant Principal Components and stratifying by self-reported race (White / Black). RESULTS/ANTICIPATED RESULTS: Of 8179 respondents passing quality filters, 6409 (78.40%) were white and 985 (12.05%) were black. Self-reported glaucoma prevalence was 7.85% and 16.34% in white and black respondents, respectively. White respondents had a mean age of 76.97 (SD 7.53) and were 57.25% female. Black respondents had a similar mean age of 74.96 (SD 7.27) and were 62.54% female. More than 87% of both groups were assessed in 2012. Preliminary GWAS analyses did not replicate known glaucoma loci and no variants attained genome-wide significance. A suggestive variant (p<1e-05) in the black population was within 10kb of a known locus, rs1196998. Future analyses will evaluate genetic association with combinations of glaucoma and comorbidities. DISCUSSION/SIGNIFICANCE OF IMPACT: Glaucoma risk is higher in minority groups than in whites, and the majority of reported genetic studies of glaucoma have been performed in individuals of European descent. It is imperative to better understand the role of genetics, environment, and health behavior in glaucoma risk. Further, understanding common mechanisms underlying diseases that co-occur with glaucoma could illuminate novel disease mechanisms that can be targeted for early intervention and/or treatment.