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/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/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/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: 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.

OBJECTIVES/GOALS: Our objective was to investigate racial differences in experiencing multiple categories of childhood trauma (CT) and the differential impact on alcohol use in individuals with alcohol use disorder (AUD). We hypothesized that there would be a differential additive effect of CT categories endorsed and drinking behaviors between racial groups. METHODS/STUDY POPULATION: Participants were recruited through the NIAAA screening protocol where they completed alcohol-related assessments including a 90-day Timeline Followback (TLFB) and the Alcohol Use Disorder Identification Test (AUDIT). Structured Clinical Interviews for DSM disorders were conducted to identify participants with lifetime alcohol dependence (DSM-IV) or AUD (DSM-5) (N = 1152). Participants self-identified as Black or White completed the Childhood Trauma Questionnaire (CTQ) which assesses 5 types of CT: emotional abuse, physical abuse, sexual abuse, physical neglect, and emotional neglect, and were classified into 3 CT groups: no trauma, 1 type of trauma, and 2+ types of trauma endorsed. RESULTS/ANTICIPATED RESULTS: For Black participants (N = 583), 21.6% experienced no trauma, 21% experienced 1 type, and 57.4% experienced 2 or more types, with the most common being physical abuse and emotional neglect. For White participants (N = 569), 32.1% experienced no trauma, 20.6% experienced 1 type, and 47.3% experienced 2 or more types, with the most common being emotional neglect and emotional abuse. There were significant associations between CT groups, TLFB, and AUDIT measures. For Black participants, AUDIT-Harm and AUDIT Total were significantly different across the 3 CT groups (all p values <0.05). For White participants, Heavy Drinking Days was significantly different across the 3 CT groups (p = 0.028), with trends for AUDIT-Harm (p = 0.061) and AUDIT-Dependence (p<0.065). DISCUSSION/SIGNIFICANCE OF IMPACT: In individuals with AUD, there were significant positive associations between the number of CT categories endorsed and alcohol use across race, suggesting a cumulative effect of CT on risky alcohol use. Future work includes exploring personality and behavioral mediators of the relationship between cumulative trauma load and drinking.

OBJECTIVES/GOALS: Our goal is three-fold: (1) to enhance protections for research participants in decentralized clinical trials (DCT) by (2) identifying the ethical and regulatory challenges posed by DCT and (3) considering possible solutions to the ethical and regulatory challenges of DCT. METHODS/STUDY POPULATION: A literature review was conducted to identify the ethical and regulatory challenges of DCT. The review showed that, with few exceptions, the publications on DCT have been written by IT experts, researchers or representatives of the pharma industry. There are hardly any independent or multidisciplinary analyses of DCT, e.g., by ethicists, medical sociologists or patients. This suggests that, currently, the potential advantages of digitized clinical research are still to be tested empirically and confirmed independently. Ethical analyses of DCT are particularly scant, which points to a gap in the current understanding of the ethical and regulatory implications of DCT and the issues they raise. Our research used the method of ethical analysis to examine the ethical challenges and to consider possible solutions. RESULTS/ANTICIPATED RESULTS: We identify the following ethical challenges of DCT: (1) challenges related to modifications of research protocols to allow for decentralized activities; (2) lack of expertise for ethical review and approval of digital tools; (3) different regulatory standards of privacy for clinical trials and for commercial digital tools, (4) risks to participants privacy and confidentiality involving both data and physical privacy; (5) the impact of the digital divide on DCT, (6) technology-related subject selection bias and marginalization of vulnerable groups, (7) imposing new burdens on research subjects and caregivers due to technology needs and smaller research teams, (8) restricted access to the study team, and (9) impact of digital technology on informed consent and participant understanding. DISCUSSION/SIGNIFICANCE: DCT became the primary form of clinical research during the COVID-19 pandemic. DCT rely on a wide range of internet-based tools for recruitment, informed consent, data collection, health monitoring, and communication with participants. The tools pose ethical and regulatory challenges, which should be addressed to ensure participant well-being.

OBJECTIVES/SPECIFIC AIMS: Sudden death in the young (SDY) occurs in people between 1 and 40 years of age who do not have a known premortem risk factor for early death. Cardiovascular diseases account for the majority of causes of SDY. Sequencing of genes associated with congenital arrhythmia susceptibility and familial cardiomyopathy reveals pathogenic variants in 30% of postmortem cases (often called “molecular autopsy”). However, better data are needed to determine the prevalence of phenotype and genotype abnormalities in surviving relatives. METHODS/STUDY POPULATION: A retrospective cohort study was performed at a tertiary pediatric center including all subjects with a family history of SDY. Cases were identified using ICD-9 codes (798.1 or .9, V17.41, V17.49, V19.8, V61.07), search of cardiology databases, and by recursive identification of all family members of a subject. Phenotype data was independently reviewed by a pediatric cardiologist. Genotype results were available when obtained by the original treating physician. RESULTS/ANTICIPATED RESULTS: Cardiac evaluations were performed in 279 subjects from 175 families, of whom 117 subjects (42%) were first-degree relatives of the proband. Mean age of the subject at time of evaluation was 9 years (SD 5.9). Most probands were over 18 years at the time of SDY: 1–4 years of age (9%); 5–12 (5%); 13–17 (16%); 18–24 (18%); 25–40 (42%). A final diagnosis was determined in 55 families (20%), and a variant in a gene potentially causative of SDY was discovered in 20/55 (36%) of those families. Variants were classified as 50% pathogenic/likely pathogenic, 50% variants of unknown significance. Cardiac testing (ECG, echo, EST, signal averaged ECG, cardiac MRI, or EP study) was abnormal in 124/279 subjects (44%). Among those with abnormal studies, 57/124 (46%) were from a family where a final diagnosis could be determined (LQT 43%, HCM 21%, ARVC 4%, other cardiomyopathy 19%, WPW 5%, CPVT 2%). However, 67/279 of total subjects (24%) had at least 1 abnormal study and a final diagnosis was not determined in the family. DISCUSSION/SIGNIFICANCE OF IMPACT: An abnormal phenotype is common among relatives referred for cardiac evaluation after SDY. While testing identifies a family diagnosis in 20% of families, many patients have abnormal cardiac testing and no clear diagnosis can be made. An improved postmortem protocol for phenotype testing in relatives of a SDY victim and improved postmortem genetic testing may lead to a higher diagnosis rate and improved risk determination in surviving family members.

OBJECTIVES/GOALS: To truly improve health equity and accessibility, we must develop a diverse and inclusive workforce. The BUILD EXITO program developed as a collaboration between a network of undergraduate programs and a CTSA hub and now has become a sustainable resource that will outlive NIH funding. We will disseminate our successful model. METHODS/STUDY POPULATION: The BUILD EXITO program has completed 10 years of NIH funding, a partnership between OCTRI and Portland State University (PSU), to create a new model of research training for underrepresented and disadvantaged students. This model provides an opportunity to learn about clinical and translational research academic careers; participate in a research enhancement and professional development curriculum;have a long-term authentic research experience; and receive enhanced mentorship. BUILD EXITO includes PSU, and local and 3 US Pacific territory 2-year colleges. We have developed a sustainable plan that includes these core elements after NIH support for the program ends. We have tracked long-term student outcomes for entry into graduate programs and the research workforce. RESULTS/ANTICIPATED RESULTS: We will describe the experimental model and the network of university and community colleges in BUILD EXITO, including PSU, U of Alaska, and colleges in US territories of Guam, Northern Mariana Islands, and American Samoa. All these universities and colleges have high proportions of underrepresented and disadvantaged students. We will present data on characteristics of the >600 students who have participated in BUILD EXITO to demonstrate the diversity of the cohort. We will also describe 4-year degree completion, engagement in the research workforce, and entry into graduate or professional programs. We will show how this has positively affected faculty inclusion of students in research, institutional policies at the 2-year and 4-year programs, and how this model has become sustainable. DISCUSSION/SIGNIFICANCE: The BUILD EXITO program developed as a collaboration of the CTSA hub at OHSU and a highly diverse undergraduate programs. We have developed a successful model for training a diverse research workforce and will disseminate this sustainable model.

Strategies are needed to ensure greater participation of underrepresented groups in diabetes research. We examined the impact of a remote study protocol on enrollment in diabetes research, specifically the Pre-NDPP clinical trial. Recruitment was conducted among 2807 diverse patients in a safety-net healthcare system. Results indicated three-fold greater odds of enrolling in remote versus in-person protocols (AOR 2.90; P < 0.001 [95% CI 2.29–3.67]). Priority populations with significantly higher enrollment included Latinx and Black individuals, Spanish speakers, and individuals who had Medicaid or were uninsured. A remote study design may promote overall recruitment into clinical trials, while effectively supporting enrollment of underrepresented groups.

The emphasis on team science in clinical and translational research increases the importance of collaborative biostatisticians (CBs) in healthcare. Adequate training and development of CBs ensure appropriate conduct of robust and meaningful research and, therefore, should be considered as a high-priority focus for biostatistics groups. Comprehensive training enhances clinical and translational research by facilitating more productive and efficient collaborations. While many graduate programs in Biostatistics and Epidemiology include training in research collaboration, it is often limited in scope and duration. Therefore, additional training is often required once a CB is hired into a full-time position. This article presents a comprehensive CB training strategy that can be adapted to any collaborative biostatistics group. This strategy follows a roadmap of the biostatistics collaboration process, which is also presented. A TIE approach (Teach the necessary skills, monitor the Implementation of these skills, and Evaluate the proficiency of these skills) was developed to support the adoption of key principles. The training strategy also incorporates a “train the trainer” approach to enable CBs who have successfully completed training to train new staff or faculty.

Translational research should examine racism and bias and improve health equity. We designed and implemented a course for the Master of Science in Clinical Investigation program of the Northwestern University Clinical and Translational Sciences Institute. We describe curriculum development, content, outcomes, and revisions involving 36 students in 2 years of “Anti-Racist Strategies for Clinical and Translational Science.” Ninety-six percent of students reported they would recommend the course. Many reported changes in research approaches based on course content. A course designed to teach anti-racist research design is feasible and has a positive short-term impact on learners.

OBJECTIVES/SPECIFIC AIMS: To assess the association between probable OSA and the sudden unexpected death in epilepsy (SUDEP-7) risk profiling index in monitored adult inpatients with epilepsy. METHODS/STUDY POPULATION: We analyzed 49 consecutive adults (>18 years) with refractory epilepsy admitted to our inpatient epilepsy monitoring unit. The SUDEP-7 inventory was performed for all subjects. Probable OSA was identified using overnight oximetry, the Sleep Apnea Sleep Disorder Questionnaire (SA-SDQ), and STOP-BANG inventory. RESULTS/ANTICIPATED RESULTS: Thirty-nine percent of participants screened positive for probable sleep apnea. Patients with high SUDEP-7 scores were more likely to have a positive screen for OSA. DISCUSSION/SIGNIFICANCE OF IMPACT: OSA is an independent risk factor for sudden cardiac death. OSA may be a hitherto unrecognized contributor to sudden death risk in epilepsy. Further studies determining the relationship between OSA, neural circulatory control and SUDEP are warranted.

Objectives/Goals: Physical activity (PA) is a well-documented protective factor against many cardiovascular diseases. PA guidelines to reduce these risks and the impact of variability are unclear, and most studies only examine a 7-day activity window. This study aimed to examine factors related to variability in step counts in a 3-year study of adults aged ≥18 years. Methods/Study Population: Included were 6,525 participants from the Michigan Predictive Ability and Clinical Trajectories study, a prospective cohort of community-dwelling adults enrolled between 8/14/2018 and 12/19/2019 who received care at Michigan Medicine and were followed for 3 years. Data were collected from Apple Watches provided to participants via the HealthKit. This secondary analysis included those with ≥4 valid weeks of data (≥4 days with at least 8 hours of wear time). Season was defined as Spring (March 20–June 20), Summer (June 21–September 21), Fall (September 22–December 20), and Winter (December 21–March 19). GEE models against the outcome of variability, defined as weekly standard deviation of step counts, and the predictor of season were adjusted for age, sex, race/ethnicity, weekly average step count, diabetes, and body mass index. Results/Anticipated Results: The average (standard deviation (SD) step counts by season were 7101 (3434) in Spring, 7263 (3354) in Summer, 6863 (3236) in Fall, and 6555 (3211) in Winter. Compared to winter, there was statistically significantly higher variability in all other seasons (p Discussion/Significance of Impact: In this cohort of community-dwelling adults, we found significant differences in variability of physical activity by season, age, and BMI. Future work will examine how this variability impacts the risk of development of cardiovascular disease, incorporating the impact and recovery trajectories of COVID-19 and other acute respiratory infections.

OBJECTIVES/GOALS: Early pediatric intensive care unit (PICU) mobilization is safe and associated with improved outcomes. Nursing-specific mobility training is desired and improves mobilization compliance. Thus, our aim is to implement a nurse-targeted, simulation-based early mobility curriculum to determine if it increases the frequency of PICU mobilizations. METHODS/STUDY POPULATION: We will conduct a single center pre-post interventional study of an in situ nurse-targeted, simulation-based early mobility curriculum. We will prospectively evaluate mobilization events in 100 patients admitted during the pre-intervention phase (n=50) and the post-intervention phase (n=50). Inclusion criteria are children ages 1 day to 17 years old admitted to the PICU for ≥3 days. Exclusion criteria include specific mobility contraindications. PICU-wide deployment will be complete when >80% of nurses have participated in the curriculum. Demographic and clinical information will be obtained. Mobility data obtained will include number of nursing-led mobilizations, highest level of mobility achieved, and potential safety events. Data will be collected from the EMR and the nurse caring for the patient. RESULTS/ANTICIPATED RESULTS: The primary endpoint will be the change in the number of nursing-led mobilization events per patient day. The secondary effectiveness outcome is the highest level of mobility achieved by patients during mobilization events in a day. A final secondary end point will be safety events defined as unplanned extubations, medical device dislodgement, falls, and cardiac arrests. Descriptive statistics for continuous variables will be presented as the median and interquartile range and categorical variables will be expressed as percentages. The effect of the simulation curriculum on the clinical outcomes will be assessed using mixed-effects models. Due to the lack of normality in number of nurse-led mobilizations and highest level of mobility achieved, the analysis will be performed using log-normal models. DISCUSSION/SIGNIFICANCE: We hypothesize that we will demonstrate the crucial importance of hands-on nursing education to improve and increase early mobility of critically ill children

OBJECTIVES/GOALS: Early life pain/stress impacts infants’ neurodevelopmental outcomes. Mitochondrial dysfunction may interface between infants’ stress and neurodevelopment. The study aims to investigate the associations between pain/stress, proteins associated with mitochondrial dysfunction, and neurobehavioral responses in preterm infants. METHODS/STUDY POPULATION: A prospective cohort study was conducted with 33 preterm infants enrolled between September 2017 and July 2022 at two affiliated NICUs in Hartford and Farmington, CT. Daily pain/stress experienced during NICU was documented. At 36-38 weeks post-menstrual age (PMA), neurobehavioral outcomes were evaluated using the NICU Network Neurobehavioral Scale (NNNS) and buccal swabs for Mass spectrometry-based proteomics analysis. Lasso statistical methods were conducted to study the association between protein abundance and infants’ NNNS summary scores. Multiple linear regression and Gene Ontology (GO) enrichment analyses were performed to examine how clinical characteristics and neurodevelopmental outcomes may be associated with protein levels and underlying molecular pathways. RESULTS/ANTICIPATED RESULTS: During NICU hospitalization, preterm premature rupture of membrane (PPROM) was negatively associated with neurobehavioral outcomes. The protein functions, including leptin receptor binding activity, glutathione disulfide oxidoreductase activity, and response to oxidative stress, lipid metabolism, phosphate, and proton transmembrane transporter activity, were negatively associated with neurobehavioral outcomes. In contrast, cytoskeletal regulation, epithelial barrier, and protection function were found to be positively associated with neurodevelopmental outcomes. In addition, mitochondrial dysfunction-related proteins (SPRR2A, PAIP1, S100A3, MT-CO2, PiC, GLRX, PHB2, and BNIPL-2, ABLIM1, UNC45A, Keratins, MUC1, and CYB5B) were found to be associated with neurobehavioral outcomes. DISCUSSION/SIGNIFICANCE: Mitochondrial dysfunction-related proteins were observed to be associated with early life pain/stress and neurodevelopmental outcomes in infants. Buccal proteins could be used to predict potential neurobehavioral outcomes. In addition, individualized skin integrity protection should be provided to preterm infants during their NICU stay.