OBJECTIVES/GOALS: In November 2020, Tufts CTSI launched a new, upgraded learning management platform for disseminating self-paced online professional education for the CTR workforce. This poster will present a case study of the first two full years of Tufts CTSI I LEARN usage to illustrate how a local and national CTR workforce engages in free online training. METHODS/STUDY POPULATION: This study will employ a retrospective analysis of 2021 and 2022 learner usage data from the Tufts CTSI I LEARN platform. The population are individuals who have engaged in one or more courses over the specified time period, where engagement is defined as having viewed at least one content item in a course. Descriptive statistics will be used to show total engagement and unique learners each year by course, popular courses by year, as well as geographical distribution of learners across both years. Two sample t-tests will be used to compare the mean percentage of course completion and mean time spent in courses for 2021 and 2022. The Wilcoxon Rank Sum test will be used to compare median courses taken per learner each year. Finally, descriptive statistics will be used to show returning learners vs new learners from 2021 to 2022. RESULTS/ANTICIPATED RESULTS: Data from 2021 show 723 learners (436 unique) engaged in 49 courses across the year. Course engagement ranged from a max of 135 to minimum of 1 learner (Mean = 14.8, Median = 6). Mean percent of course content viewed was 69.4 (Median = 63.7%) and mean time spent on course content was 41.3 minutes (Median = 33.6). Preliminary data from Q1-Q3 of 2022 find 557 learners (328 unique) have engaged in 69 courses. Based on this trend, it is anticipated that 2022 will match or exceed 2021 engagement levels. Recently launched courses are expected to have higher average engagement than legacy courses ported from the previous version of the platform. DISCUSSION/SIGNIFICANCE: Results from this evaluation will suggest strategies to enhance engagement in current courses and identify opportunities for future course development. Increasing engagement in self-paced CTR education is an important complement to formal workforce development and training programs.

OBJECTIVES/GOALS: To engage community partners in full spectrum of translational sciences participants will: 1. Develop community-based research projects. 2. Evaluate pilot awards submitted to the Community Advisory Board for funding. 3. Actively participate in the research cores at the CTSC METHODS/STUDY POPULATION: The Academy workshop curriculum supports an in-depth examination of the translational research process, introduction to the CTSC Community Advisory Board; as well as to community based participatory research; grant evaluation process, and the need for active community involvement in various cores, and clinical trials. Workshops presented by tenured professionals and community leaders with multi-media out of class assignments. Scheduled sessions were 1 hour, and 15 minutes. Exams given post class session with a mean score of 22.8/30 for session 1 and a mean score of 23.80/30 for session 2. RESULTS/ANTICIPATED RESULTS: The Community Research Academy (CRA) has thus far held for two sessions. Of the 45 enrollees, 20 completed the entire program. Of those, ten have joined the CAB board; and three have actively participated in the pilot award review process. One of the CRA graduates will be publishing her photo novella assignment in Health Affairs. One person changed careers,: and several people have taken all the classes for a second time. Many participants originally came from our faith-based connections. Now word of mouth is expanding the program. DISCUSSION/SIGNIFICANCE: Community engagement ensures that the fresh voices of diverse populations are involved in translational science. Their input ultimately leads to creating novel clinical innovations; such community-driven ingenuity which addresses the deeply felt needs of those communities.

Objectives/Goals: Poor visual memory and perceptual organization task performance predicts cognitive decline and is sensitive to dementia severity. No genome-wide association study (GWAS) has assessed the genomic basis of cognitive visual-spatial phenotypes. We aimed to identify common genetic variants associated with visual memory and spatial organization. Methods/Study Population: We included dementia- and stroke-free participants aged 45 years or older from up to seven cohorts in the Cohorts for Heart and Aging Research in Genomic Epidemiology (CHARGE) consortium, who performed cognitive tasks assessing delayed visual memory (e.g., Benton Visual Retention Test (BVRT, n = 10,934) and visual reproductions (VR, n = 5,527)) or spatial organization (i.e., Hooper Visual Organization Test (HVOT, n = 5,024)). Each cohort used linear regression models to relate common genetic variants imputed to the 1000 Genomes panel to each cognitive phenotype, adjusting for age, sex, population stratification, and education. Summary statistics for the BVRT were meta-analyzed using METAL. Combined GWAS was used for a joint analysis of all traits. Results/Anticipated Results: We identified a genome-wide significant variant related to BVRT performance located near the TSHZ3 gene (rs10425277, p = 6.76×10–9). TSHZ3 is important for the development and function of cortical projecting neurons and may be implicated in Alzheimer’s disease progression by repressing CASP4 transcription. Multitrait analyses, including BVRT, VR, and HVOT, identified two additional variants of interest in SMYD3 gene (rs10802275, p = 5.58×10–7) and near ZFPM2 (rs2957459, p = 2.03×10–7), both of which are overexpressed in the brain and have important implications for neurodevelopment. SMYD3 may be directly involved in synaptic dysfunction and has been shown to be upregulated in the prefrontal cortex of Alzheimer’s disease patients. Discussion/Significance of Impact: Our findings suggest that variants related to visual memory and spatial organization are involved in neurodevelopmental and degenerative pathways. This GWAS adds to the growing body of GWAS literature on the genetic basis of cognitive function. Additional analyses are underway to replicate these findings and extend functional annotation.

OBJECTIVES/SPECIFIC AIMS: Triple-negative breast cancer (TNBC) accounts for one-fifth of the breast cancer patient population. The heterogeneous nature of TNBC and lack of options for targeted therapy make its treatment a constant challenge. The co-deficiency of tumor suppressors p53 and ARF is a significant genetic signature enriched in TNBC, but it is not yet clear how TNBC is regulated by this genetic alteration. METHODS/STUDY POPULATION: To answer this question, we established p53/ARF-defective murine embryonic fibroblast (MEF) to study the molecular and phenotypic consequences in vitro. Moreover, transgenic mice were generated to investigate the effect of p53/ARF deficiency on mammary tumor development in vivo. RESULTS/ANTICIPATED RESULTS: Increased transformation capability was observed in p53/ARF-defective cells, and formation of aggressive mammary tumors was also seen in p53-/-ARF-/- mice. RNA-editing enzyme ADAR1 was identified as a potential mediator for the elevated oncogenic potential. Interestingly, we found that the overexpression of ADAR1 is also prevalent in human TNBC cell lines and patient specimen. Using short hairpin RNA (shRNA) to reduce ADAR1 expression abrogated the oncogenic potential of human TNBC cell lines, while non-TNBC cells are less susceptible. Different levels of RNA editing of known ADAR1 targets were detected in shRNA-treated human TNBC cell lines, suggesting that ADAR1-mediated RNA editing contributes to TNBC pathogenesis. DISCUSSION/SIGNIFICANCE OF IMPACT: These results indicate critical roles played by the tumor suppressors p53 and ARF in the pathogenesis of TNBC, partially through affecting ADAR1-mediated RNA editing. Further understanding of this pathway could shed light on potential vulnerabilities of TNBC and inform the development of personalized therapies based on patients’ genetic signiatures.

OBJECTIVES/GOALS: We investigated hemodynamic measures in young black and white adults below the age of 50 years to identify mechanisms that may predispose blacks to more CVE. METHODS/STUDY POPULATION: We recruited 276 young blacks and white adults in Jackson, MS (mean age: 33 ± 9 years; 70% women; 57% Black;). Participants had clinical and vascular tonometry parameters obtained. Vascular measures included carotid femoral pulse wave velocity, central and peripheral pulse pressure (CPP and PPP), characteristic impedance (Zc), augmentation index, and forward pressure wave. Race-specific characteristic means and proportions were tested using Student’s T- or Chi-square tests. After further sex stratification, significant variation in characteristics among sex within race were tested using nested ANOVA. RESULTS/ANTICIPATED RESULTS: Characteristics of the study group stratified by race were found to be similar. Vascular measures stratified by race revealed blacks to have significantly higher Zc (p = 0.03) and PPP (p = 0.03) than whites. DISCUSSION/SIGNIFICANCE OF IMPACT: In this study of vascular hemodynamics in young black and white participants we found differences in Zc and PPP. Findings suggest that differing relations between proximal aortic diameter and wall stiffness may contribute to the racial disparity in CVE in adults. This finding could offer an explanation to the beneficial effects of treatment modalities that target aortic stiffness in blacks.

ABSTRACT IMPACT: This change will improve primary care physicians and pediatrics ability to identify, intervene and prevent obesity related renal damage in the vulnerable population of young adults OBJECTIVES/GOALS: Obesity related glomerulopathy has a reversible stage manifested as hyperfiltration. Early intervention depends on the ability to identify hyperfiltration. Hyperfiltration prevalence is underestimated using the currently recommended formula We investigated whether calculating BSA-adjusted GFR will more readily identify hyperfiltration. METHODS/STUDY POPULATION: We extracted data from a large urban, multi-institutional Electronic Health Records (EHR) clinical data research network to construct an EHR data base of 60,549 women and girls ages 12-21 years from the New York metropolitan area. EGFR was calculated in two ways, 1) according to age appropriate formula, and 2) according to age appropriate formula and adjusted to body surface area (BSA). BMI-for-age values were classified according to the World Health Organization schema and grouped according to the CDC definitions. BSA was calculated according to the Du-Bois formula. Hyperfiltration was defined by a threshold of 135ml/min. The Bland Altman method assessed the agreement between formulas across the different BMI groups. RESULTS/ANTICIPATED RESULTS: Serum creatinine values were similar across different BMI groups. Comparing eGFR values, hyperfiltration rates were similar across BMI groups, ranging between 4%-6.6%. BSA-adjusted GFR was different across BMI groups: hyperfiltration rates were 0.81% for the underweight group, 2.56% for the normal weight, 12.18% for the overweight and 39% in the obese group. This trend of hyperfiltration paralleled the the rise in urine creatinine across BMI groups. DISCUSSION/SIGNIFICANCE OF FINDINGS: BSA-adjusted GFR more sensitively detects hyperfiltration due to obesity than does eGFR. Calculating BSA-adjusted GFR will improve primary care and pediatric physicians’ ability to identify, intervene and prevent early ORG. Changes in body composition may account for the increasing discordance between BSA-adjusted and eGFR as BMI rises.

Story is the oldest known way of sharing knowledge and information and engages us in our collective humanity. In research settings, story brings meaning to complex ideas, making them feel palpable and connects us with our audience. Historically, the disciplines that take a research interest in the importance of narrative have been largely in fields like the philosophy of science/medicine, medical humanities, and sociology though story is “always already” a part of scientific research. Humanities have gained traction in medical and science education, and researchers are seeking such curricula to communicate more effectively with the public and their students. We believe that story is an effective tool to enable CTS investigators to be effective educators and communicators of translational science. Story-based interdisciplinary pedagogy emphasizes an approach encouraging clinical researchers to keep the human story as the driving force of research design, dissemination, and application of research to diverse audiences. In this article, we provide backgrounds on successful programs that have used story in science communication and education as well as a tool researchers can use to incorporate the structure of story into their own work.

Extant literature reveals how patients of marginalized social identities, socioeconomic status (SES), and medical experiences – especially patients of color and older adults – are underrepresented in cancer clinical trials (CCTs). Emerging evidence increasingly indicates CCT underrepresentation among patients of lower SES or rural origin, sexual and gender minorities, and patients with comorbid disability. This review applies an intersectional perspective to characterizing CCT representativeness across race and ethnicity, age, sexual and gender identity, SES, and disability. Four databases were systematically queried for articles addressing CCT participation inequities across these marginalizing indicators, using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. One hundred one articles were included in a qualitative evaluation of CCT representativeness within each target population in the context of their intersectional impacts on participation. Findings corroborate strong evidence of CCT underrepresentation among patients of color, older age, lower SES, rural origin, and comorbid disabling conditions while highlighting systemic limitations in data available to characterize representativeness. Results emphasize how observed inequities interactively manifest through the compounding effects of minoritized social identity, inequitable economic conditions, and marginalizing medical experiences. Recommendations are discussed to more accurately quantify CCT participation inequities across underserved cancer populations and understand their underpinning mechanisms.

OBJECTIVES/SPECIFIC AIMS: The goal of this study is to further evaluate underlying disease parameters in respiratory syncytial virus (RSV) infection, that is reduction in antioxidant potential, and determining if supplementation of the antioxidant enzyme catalase could be employed as a potential therapeutic. METHODS/STUDY POPULATION: Nasopharyngeal secretions were obtained from patients (<2 years old) verified for RSV infection, and assessed for catalase activity and correlated with disease parameters. In addition, the BALB/c animal model of RSV infection was utilized to directly study the effect of supplemental catalase on RSV-related disease parameters in vivo. The catalase formulation used in these studies is pegylated, and has been tested to provide long-term increased catalase activity in vivo. We are also currently working on designing an in vitro model of catalase supplementation in A549 bronchial epithelial cells. RESULTS/ANTICIPATED RESULTS: Our preliminary data shows that patients with more severe disease (based on hospitalization, oxygen supplementation) have significantly lower levels of catalase activity (p<0.02). Additionally, when pegylated-Catalase (PG-CAT) treatment is utilized in RSV infection of mice, there is significant improvement in several disease parameters. PG-CAT-treated mice show an attenuated body weight loss (p<0.001) and clinical disease (p<0.02), and also have lower levels of key pro-inflammatory cytokines including CXCL1 and TNF-α. PG-CAT treatment also resulted in a minor decrease in viral titer, which is being further evaluated. In addition, PG-CAT treatment resulted in an improvement in airway hyperresponsiveness observed at baseline, we are further characterizing this improvement and also conducting methacholine challenges. Currently, we are working to determine the underlying mechanism through which PG-CAT results in these improvements, and whether it is through changes in immune cell populations, cellular signaling or apoptosis signaling pathways (i.e., caspases). DISCUSSION/SIGNIFICANCE OF IMPACT: RSV is the leading cause of viral pneumonia and bronchiolitis in infants, with no vaccines or effective therapeutics available currently. Our study indicates that catalase activity could be used as a potential correlate for disease severity and be used as an indicator of disease during patient treatment. Additionally, and more importantly supplementation of catalase could be used as a potential therapeutic for treatment of RSV.

OBJECTIVES/SPECIFIC AIMS: Despite aggressive chemotherapy, surgical resection, and radiation therapy, glioblastoma remains almost universally fatal. In a pilot, randomized, and blinded clinical trial, we recently demonstrated that administration of RNA-loaded DC vaccines was associated with significantly improved progression-free and overall survival in patients with glioblastoma (Mitchell et al., Nature, 2015). Furthermore, clinical outcomes correlated with DC migration to vaccine-site draining lymph nodes measured by Indium-111 labeling of RNA-loaded DCs and SPECT/CT imaging. Although these studies demonstrated that tracking DC migration may be an important clinical biomarker for response to DC vaccination, the complexity and regulatory requirements associated with nuclear labelling to track DC migration limits widespread application of this technique. We have therefore developed RNA-loaded magnetic nanoparticles (RNA-NPs) to enhance DC migration to LNs and track that migration with a widely available imaging modality (i.e., MRI). METHODS/STUDY POPULATION: Cationic liposomes were loaded with iron oxide nanoparticles with or without cholesterol. The resulting nanoparticles were complexed with RNA and used to transfect DCs ex vivo. RNA-NP-loaded DsRed+ DCs were then injected intradermally into mice and tracked noninvasively with T2-weighted 11T MRI before excision and quantification with flow cytometry. RESULTS/ANTICIPATED RESULTS: In vitro experiments demonstrate that iron oxide loading does not reduce RNA-NP-mediated transfection of DCs. Additionally, replacement of cationic lipids with cholesterol increased RNA-NP transfection of the DC2.4 cell line and enhanced the T cell stimulatory capacity of treated bone marrow-derived dendritic cells (BMDCs). Compared to electroporation, RNA-NPs enhanced DC migration to lymph nodes and reduced T2 MRI intensity in DC-bearing lymph nodes. DISCUSSION/SIGNIFICANCE OF IMPACT: This data suggests that iron oxide-loaded RNA-NPs enable noninvasive cell tracking with MRI and enhance DC migration to lymph nodes. We have further shown that inclusion of cholesterol in RNA-NPs augments the stimulatory capacity of transfected DCs. Future work will consider effects of RNA-NPs on antitumor immune responses and the utility of MRI-detected DC migration as a biomarker of vaccine efficacy.

OBJECTIVES/SPECIFIC AIMS: This research examined 3 aims to address the need to understand and quantify exertion in infants. Aim 1: Develop a schema to identify and code exertional behaviors in infants during treadmill stepping. Aim 2: Establish feasibility for the schema’s use with clinical populations. Aim 3: Pilot the schema in a study designed to induce infant exertion. METHODS/STUDY POPULATION: Aims 1 and 2 were achieved using existing treadmill stepping data. The data used in Aim 1 included eight typically-developing infants (age 7-10 months) who were able to sit independently, but not walk. The data used in Aim 2 came from two separate data sets from infants who took more than 10 steps in a 30-second trial: Data set A included six typically-developing infants (age 2-5 months) who were unable to sit independently (developmentally comparable to atypical populations who might receive treadmill interventions). Data set B included six infants with Spina Bifida (age 3-10 months). Aim 3 was addressed with a prospective study using an exertion model. Pre-walking, typically developing infants (age 8-10 months) underwent five total stepping trials. Trial 1 determined the infant’s individualized maximum stepping speed; trials 2-5 were each 60 seconds and alternated between a baseline stepping speed of.20 m/s and the infant’s maximum stepping speed determined in trial 1. All video data were coded for step type, step frequency, and exertional behavior. RESULTS/ANTICIPATED RESULTS: Aim 1: Two behaviors were identified and determined to capture infant exertion: foot dragging and leg crossing. Aim 2: The feasibility of capturing exertion with these two behaviors was established for young infants and infants with neuromotor delays, with exertional behaviors increasing with stepping exposure (p< 0.05). Aim 3: Total exertion (foot dragging + leg crossing) was higher in the maximum speed trials compared to baseline trials (p = 0.005). DISCUSSION/SIGNIFICANCE OF IMPACT: Exertion in infants can be quantified. The exertion schema developed with this study will support the development of dosing guidelines for infant treadmill intervention. The next step in this line of research is to examine the correlation between infant exertion and heart rate, in effort to move from behaviorally-informed protocols to more precise, individualized protocols based on the physiological response of the infant.

OBJECTIVES/GOALS: Alachua TranQuility was formed in early 2016 to address needs of transgender people in Alachua County, Florida and surrounding areas. Increasing awareness of gender identity issues created a growing population in need of additional health and social services to counter the negative health outcomes traditionally experienced by transgender people. METHODS/STUDY POPULATION: The group met monthly at UF HealthStreet, a CTSA community engagement program, where social and medical referrals and opportunities to participate in research were made available to attendees. Those attending included transgender and gender non-conforming individuals as well as family, friends and other allies. Outreach was conducted through social media, physical fliers, and tabling at community events. TranQuility partnered with local LGBTQ organizations to promote activities. Leadership was provided by a steering committee that changed over time, with feedback from attendees guiding planning. Simultaneous to TranQuilitys founding, a Youth Gender Clinic was established at UF Health, clearly demonstrating the need for services aimed at the transgender population. RESULTS/ANTICIPATED RESULTS: Between February 2016 and September 2019, attendance at meetings was logged per HealthStreet protocols. Age of attendees ranged from 10 to 75, and most were non-Hispanic whites. More than 300 individuals attended meetings across time, and many attended multiple meetings, with greater than 15% having attended five or more meetings. Attendance averaged around 30+ each month, with a high of 76. Educational programming was offered, such as information on hormone therapy and name/gender marker change, and a parent support group was formed to meet concurrently with the main group. Social opportunities were very popular, as a safe place for people to present as their authentic selves. The coronavirus pandemic curtailed in-person meetings at HealthStreet in 2020, but the group plans to return to this format in 2022. DISCUSSION/SIGNIFICANCE: TranQuility has become an established organization to which other groups provide referrals. Qualitatively it is clear that many people have been positively impacted. An ongoing discussion for the group, however, has been to improve outreach to transgender people of color, who typically suffer from the most discrimination and anti-trans violence.

OBJECTIVES/SPECIFIC AIMS: (1) Characterize the prevalence and initial pharmacological management of chronic pain syndromes during pregnancy in a women’s mental health program. (2) Describe the severity and qualitative characteristics of chronic pain during pregnancy and the acute postpartum period. (3) Compare obstetrical and neonatal outcomes between pregnant women with and without chronic pain syndromes. METHODS/STUDY POPULATION: A chart review was conducted to identify all pregnant women who presented for an initial evaluation to the Women’s Mental Health Program (WMHP) at the University of Arkansas for Medical Sciences from July 2013 to June 2016. We excluded respondents <18 years of age or who did not consent to having their information used for research purposes. Demographic information, past and current medical histories, and medication history were obtained from written and electronic medical records. Chronic pain complaints and medication history are presented as counts and percentages. In an ongoing prospective, longitudinal study of pregnant women with chronic pain, women are enrolled before 20 weeks gestation and followed throughout pregnancy and the first 3 months postpartum. Study visits occur at 4-week intervals; and pain characteristics, analgesic exposures, other medications, and depressive measures are collected. Obstetrical and neonatal outcomes are obtained following delivery. Subjects will be compared based on pain types (ie, neuropathic pain, non-neuropathic pain, and controls) and treatment exposures (eg, +/− opioids). Primary outcome measures include visual analog scale (VAS). Secondary outcome measures include other pain and depression assessments. Data will be analyzed using SAS 9.4. A p-value of<0.05 was considered statistically significant. RESULTS/ANTICIPATED RESULTS: (1) Chronic pain conditions were reported by 28.2% (44/156) of the initial referrals to the WMHP. (2) 95.5% of respondents with chronic pain were taking at least 1 medication, and 59.5% were taking 2 or more medications. Mean number of medications used were 2.6±2.1.3. The most common medications reported were acetaminophen (43.2%), opioids (43.2%), and sedative/hypnotics (36.4%). Non-pharmacological therapy (eg, physical therapy and transcutaneous electrical nerve stimulation) was reported by 20.5% of respondents. (4) We anticipate that measures of pain severity will increase in pregnancy, peak in the third trimester, and decline in the postpartum period. (5) We foresee that the prospective results will confirm the chart review as indicated by a higher rate of medication exposures during pregnancy, including non-analgesic medications in the women with chronic pain syndromes. (6) We expect women with chronic pain syndromes to have a higher rate of obstetrical complications, specifically pre-term delivery and operative delivery. (7) Finally, we anticipate that chronic pain syndromes and management will result in a higher rate of neonatal complications, specifically neonatal intensive care unit admission, neonatal respiratory problems, and small for gestational age infants. DISCUSSION/SIGNIFICANCE OF IMPACT: Chronic pain syndromes are prevalent in more than one-quarter of pregnant women in our study with the majority of women using pharmacological agents to manage their condition. This prevalence is greater or equal to than other common obstetrical conditions, such as gestational diabetes or preterm delivery. The novel prospective data will be germane to the clinical care of pregnant women with chronic pain disorders. Clinical practice will be better informed by our data regarding the potential impact of chronic pain and its management on pregnancy course and perinatal outcomes. These data will provide the initial foundation for the development of treatment guidelines for the management of chronic pain syndromes during the perinatal period.

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: 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: 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: 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.