OBJECTIVES/SPECIFIC AIMS: A mimic of congenital infections and a rare genetic cause of interferon overproduction, Aicardi Goutières Syndrome (AGS) results in significant neurologic disability. AGS is caused by pathogenic changes in the intracellular nucleic acid sensing machinery (TREX1, RNASEH2A, RNASEH2B, RNASEH2C, SAMHD1, ADAR1, and IFIH1). All affected individuals exhibit neurologic impairment: from mild spastic paraparesis to severe tetraparesis and global developmental delay. We hypothesize that genotype influences the heterogeneous developmental trajectory found in AGS. METHODS/STUDY POPULATION: To characterize this spectrum, age and symptoms at presentation and longitudinal developmental skill acquisition was collected from an international cohort of children (n=88) with genetically confirmed AGS. RESULTS/ANTICIPATED RESULTS: We found that individuals present at variable ages, with the largest range in SAMHD1, ADAR, and IFIH1. There are 3 clusters of symptoms at presentation: altered mental status (irritability or lethargy), systemic inflammatory symptoms, and acute neurologic symptoms, with variability across all genotypes. By creating Kaplan-Meier curves for developmental milestones, we were able to create genotype-based developmental trajectories for the children affected by the 5 most common genotypes: TREX1, IFIH1, SAMHD1, ADAR, and RNASEH2B. Individuals with AGS secondary to TREX1 were the most severely affected, significantly less likely to reach milestones compared to the other genotypes, including head control, sitting, and nonspecific mama/dada (p-value <0.005). Individuals affected by SAMHD1, IFIH1, and ADAR collectively attained the most advanced milestones, with 44% of the population achieving a minimum of a single word and 31% able to walk independently. Three retrospective scales were also applied: Gross Motor Function Classification System, Manual Ability Classification Scale, and Communication Function Classification System. Within each genotypic cohort, there was pronounced heterogeneity. DISCUSSION/SIGNIFICANCE OF IMPACT: Our results demonstrate the influence of genotype on early development, but also suggest the importance of other unidentified variables. These results underscore the need for deep phenotyping to better characterize subcohorts within the AGS population.

ABSTRACT IMPACT: This work will discuss informatics methods enabling the use of exposure health data in translational research. OBJECTIVES/GOALS: 1. Characterize gaps and formal informatics methods and approaches for enabling use of exposure health in translational research. 2. Education of informatics methods enabling use of exposure health data in translational research. METHODS/STUDY POPULATION: We performed a scoping review of selected literature from PubMed and Scopus. In addition we reviewed literature and documentation of projects using exposure health data in translation research. RESULTS/ANTICIPATED RESULTS: Primary challenges to use of exposure health data in translational research include: (1) Generation of comprehensive spatio-temporal records of exposures, (2) Integration of exposure data with other types of biomedical data, and (3) Uncertainties associated with using data as exact quantifications of exposure which are dependent on both - the proximity of measurement to subject under consideration and the capabilities of measuring devices. We identified 9 major informatics methods that enable incorporation and use of exposure health data in translational research. While there are existing and ongoing efforts in developing informatics methods for ease of incorporating exposure health in translational research, there is a need to further develop formal informatics methods and approaches. DISCUSSION/SIGNIFICANCE OF FINDINGS: Depending on the source about 50 - 75% of our health can be quantified to be a contribution of our environment and lifestyles. In this presentation, we summarize the studies and literature we identified and discuss our key findings and gaps in informatics methods and conclude by discussing how we are covering these topics in an informatics courses.

OBJECTIVES/SPECIFIC AIMS: To conduct a preliminary evaluation of the Social and Behavioral Research Best Practices Course. METHODS/STUDY POPULATION: Learners are sampled from 5 institutions: University of Michigan, University of Rochester, University of Florida, Boston University, and University of Buffalo. Learners who take the course and consent to be in the study receive a web link to a survey immediately after course completion and at 2–3 months follow up. In addition to demographic information, learners will report their perceptions of usefulness and relevance of the course to their job, their satisfaction with the course and associated job aids, and at follow-up, if and how the course impacted their work. Additional information will be collected from the learning management systems which host the course at each institution. The data collected will include the number of participants who take the course, the number who complete, how many times the course was attempted, and pass rates. RESULTS/ANTICIPATED RESULTS: We anticipate that several hundred learners will take the course by the end of our project. Of learners who agree to participate in the survey, we anticipate that they will find the course useful and relevant to social and behavioral clinical trials and will be satisfied with the course. Information including suggestions about missing content, items or content that were not extremely clear, or any other comments will be collected to iterate and expand the course. DISCUSSION/SIGNIFICANCE OF IMPACT: This course was developed to fill a gap in training in good clinical practice for social and behavioral research. An evaluation of how the training provided in the course impacts the jobs of learners is needed both to ensure that the most relevant information is included in the course as well as to identify ways that the training may contribute to the quality and safety of social and behavioral clinical trials.

OBJECTIVES/GOALS: While CTSA hubs have contributed to the successes in COVID-19 clinical research, understanding the role of the CTSA consortium in translating vaccine efficacy and availability to awareness and implementation. The goal of this study is to quantitatively assess the use of social media in the dissemination of COVID 19 vaccine content across 60+ CTSA Hubs. METHODS/STUDY POPULATION: Structured search terms in the CTSA Search Solutions database were used to identify CTSA Hub website pages highlighting “COVID-19 Vaccination Information.” Each link identified was manually reviewed for vaccination content. The links and content identified by CTSA Search Solutions were then validated by advanced Google search operator “vaccine site” and manual review of CTSA Hub websites. Official CTSA Hub social media platforms were searched for vaccine proliferation content from January 1, 2020 to November 11, 2021. Data points collected included Community Targeted Vaccine Content, Vaccine Awareness, Vaccine Distribution, Vaccine Clinical Trial, Vaccine Related Media, Social Media Presence, and Social Media Vaccine Proliferation. RESULTS/ANTICIPATED RESULTS: In examining content, of the 64 listed CTSA Hubs, 52 of (67.2%) hosted one or more categories of COVID-19 vaccine information, and 16 (25%) hosted three or more categories. The most common category was “Community Targeted Information” with 27 (42.2%) hubs; the least common category was “Vaccine Distribution” with 5 (7.8%) hubs. Examining social media for vaccine proliferation, 41 (64.1%) hubs had connected social media and 23 (35.9%) hubs had vaccine proliferation posts. The most common platform was Twitter with 37 hubs; the least common platform was Instagram with 5 hubs. DISCUSSION/SIGNIFICANCE: Our investigation demonstrated varying Phase 4 translational efforts via social media among the CTSA Hubs for COVID-19 vaccine implementation. These novel findings identify promising opportunities for enhancement while emphasizing proven strategies from CTSA hubs nationwide. Further research will elucidate granular trends among CTSA hubs.

OBJECTIVES/SPECIFIC AIMS: HIV is a chronic disease that affects the immune system. HIV+ people live more thanks to effective antiretroviral treatments. The scientific data demonstrate that HIV+ is associated to the cognitive impairment presented in the 50% of the patient. The objective of this study is to determine the correlation between emotional dysfunction and perceived stigma in HIV+ women and its effects HIV-associated neurocognitive disorders (HAND). METHODS/STUDY POPULATION: HIV+ women will be recruited form the Hispanic Longitudinal Cohort and evaluated questionnaires for emotional dysfunction and stigma, neuropsychological tests, and MRI. RESULTS/ANTICIPATED RESULTS: We anticipated that women with HIV+ will experience higher levels of emotional dysfunction (ie, fear) and perceived stigma when compared with the control group. Women with HIV infection will present an association between emotional dysfunction most like fear and perceived stigma when compared with the control group. This correlation will be associated with HAND. The women with HIV infection will present circuit integrity dysfunction associated with emotional dysfunction and perceived stigma as determined by DTI and connectivity (MRI). DISCUSSION/SIGNIFICANCE OF IMPACT: HIV stigma and emotional dysfunction have a negative impact in quality of life (QOL). This effect can be improved with several treatment interventions with eventual improvement in adherence, emotional control, and QOL.

OBJECTIVES/GOALS: The goal of this study is to explore the effectiveness of a digital health education curriculum aimed at increasing syphilis beliefs and screening behavior among young African American females residing in Jefferson County, AL. METHODS/STUDY POPULATION: Study Population: African American, cis-gender women, aged 18-29, who reside in Jefferson County, AL Sample Size: N = 50 Measurements: 1) Champion Health Belief Model scale adapted for measuring syphilis perceptions and syphilis screening behavior Analysis Plan: IBM SPSS Statistics 29 will be used to analyze intervention data. Pre-and post-intervention data will be analyzed to calculate a screening request rate for each period. We will use Cohen’s D effect sizes to measure screening requests rate differences post intervention completion. The small sample size and exploratory nature of this project make the descriptive statistics valuable. RESULTS/ANTICIPATED RESULTS: We anticipate that the digital syphilis prevention intervention will positively influence participants’ beliefs regarding syphilis and their syphilis screening behaviors. DISCUSSION/SIGNIFICANCE: Innovative preventive approaches to reducing syphilis disease spread is a national priority as rates have risen annually throughout the last decade. African American women experience the greatest disease burdens associated with syphilis. This study leverages tailored strategies to effectively address this ailment in the target population.

OBJECTIVES/SPECIFIC AIMS: Knowing how to deliver culturally responsive care is of increasing importance as the nation’s patient population diversifies. However, unless cultural competence is taught with an emphasis on self-awareness (Wear, 2007) and critical consciousness (Kumagai and Lypson, 2009) learners find this education ineffective (Beagan, 2003). This study examines how physicians perceive their own social identities (eg, race, socio-economic status, gender, sexual orientation, religion, years of experience) and how these self-perceptions influence physician’s understandings of how to practice culturally responsive care. METHODS/STUDY POPULATION: This exploratory study took place at a university in the Intermountain West. We employed a qualitative case study method to investigate how academic physicians think about their identities and approaches to clinical care and research through interviews and observations. In total, 25 participants were enrolled in our study, with efforts to recruit a diverse sample with respect to gender and race as well as years of experience and specialty. Transcriptions of interviews and observations were coded using grounded theory. One major code that emerged was defining experiences: instances where physicians reflected on both personal and professional life encounters that have influenced how they think about themselves, how they understand an aspect of their identity, or why this identity matters. RESULTS/ANTICIPATED RESULTS: Two main themes emerged from an analysis of the codes that show how physicians think about their identities and their approaches to practice. (1) Physicians with nondominant identities (women, non-White) could more easily explain what these identities mean to them than those with dominant identities (men, White). For example, women in medicine had much to say about being a woman in medicine, but men had barely anything to say about being a man in medicine. (2) There was a positive trend between the number of defining experiences a physician encountered in life and the number of connections they made between their identities and the manner in which they practiced, both clinically and academically. It appeared that physicians who have few defining experiences made few connections between identity and practice, those with a moderate number of experiences made a moderate number of connections, and those with many experiences made many connections. Physicians who mentioned having many defining experiences were more likely to be able to articulate how those experiences were incorporated into their approaches to patient care. DISCUSSION/SIGNIFICANCE OF IMPACT: (1) According to literature in multicultural education, those with dominant identities do not think about their identities because they do not have to (Johnson, 2001). One privilege of being part of the majority is not having to think about life from a minority perspective. This helps to explain why women and non-White physicians in this study had more anecdotes to share about these identities—because they have had defining experiences that prompt reflection on these identities. (2) We propose that struggles and conflict are what compel physicians to reflect on their practice (Eva et al., 2012). Our findings suggest that physicians are more prepared to apply what they have learned from their own identity struggles in delivering culturally responsive care when they have had more opportunities to reflect on these identities and situations. Findings from this study have implications for transforming approaches to medical education. We suggest that medical education should provide learners with the opportunity to reflect on their life experience, and that providers may need explicit instruction on how to make connections between their experiences and their practice.

OBJECTIVES/GOALS: Our objective was to evaluate the FLEX Leadership Development Program for School of Medicine Women Faculty affiliated with 4 independent hospital systems throughout Northern Ohio to determine whether women faculty develop leader self and means efficacy. We also examined whether self-efficacy is sustained over time following program completion. METHODS/STUDY POPULATION: We did a prospective multiple cohort study to determine whether FLEX Program graduates develop and sustain leader self and means efficacy as measured by the Leader Efficacy Questionnaire (LEQ) (Hannah & Avolio, 2012). The LEQ assesses both leader confidence in their capabilities (self-efficacy) and the availability of sufficient external resources (means efficacy) to achieve their goals. We surveyed participants from 5 FLEX cohorts (2017-2021) using a pre-, post-, and 1-year follow-up LEQ, which allowed each participant to act as their own control subject, pre-test, and experimental post-test. The follow-up tested whether the change from pre- to post-test was sustained over time. The comparison group was non-participant women faculty from the same institutions over a similar 2-year period. RESULTS/ANTICIPATED RESULTS: Diverse FLEX graduates from 5 different cohorts showed highly significant increases in pre- to post-program leader self-efficacy which were sustained up to one year after program completion. Overall leader efficacy as well as its three component sub-constructs (action-, means- and self-regulation- efficacy) all significantly increased equally, suggesting both that the FLEX program had robust effects on its participants, and all aspects of leader efficacy improved. We observed a mildly significant decrease at 1-year follow-up in the overall LEQ, which appeared to be driven entirely by the leader means efficacy. The large comparison group of women faculty did not show any significant changes in leader self-efficacy over a comparable 2-year period. DISCUSSION/SIGNIFICANCE: FLEX confers sustainable gains in leader self-efficacy. Post-program self-efficacy decreases are driven by the leader means efficacy which measures how work environments affect their leadership. This suggests that institutions must take responsibility for making structural changes to improve the working environment for women leaders.

OBJECTIVES/SPECIFIC AIMS: MicroRNAs are small, non-coding RNAs that control gene expression by inhibiting protein translation. Preclinical studies in rodent stroke models suggest that changes in microRNA expression contribute to neural repair mechanisms. To our knowledge, no one has previously assessed microRNA changes during the recovery phase of human stroke. Our goal was to determine whether patients with significant upper limb recovery following stroke have alteration of neural repair-related microRNA expression when compared to those with poor recovery. METHODS/STUDY POPULATION: Plasma was collected at 19 days post-stroke from 27 participants with mild-moderate upper extremity impairment enrolled in the Critical Periods After Stroke Study. MicroRNA expression was assessed using TaqMan microRNA assays (Thermo Fisher Scientific). Good recovery was defined as ≥6 point change in the Action Research Arm Test (ARAT) score from baseline to 6 months. Bioinformatics analysis compared the plasma microRNA expression profiles of participants with good Versus poor recovery. Candidate biomarkers were identified after correcting for multiple comparisons using a false discovery rate <0.05. RESULTS/ANTICIPATED RESULTS: Eleven microRNAs had significantly altered expression in the good (n=22) Versus poor (n=5) recovery groups, with 2 showing increased expression—miR-371-3p and miR-520g, and 9 showing decreased expression—miR-449b, miR-519b, miR-581, miR-616, miR-892b, miR-941, miR-1179, miR-1292, and miR1296. Three of these could be implicated in neural repair mechanisms. Elevated miR-371-3p levels increase the likelihood that pluripotent stem cells will differentiate into neural progenitors. MiR-892b decreases levels of amyloid precursor protein, which has been implicated as a regulator of synapse formation. Finally miR-941, the only known human-specific microRNA, downregulates the CSPα protein which is involved in neurotransmitter release. DISCUSSION/SIGNIFICANCE OF IMPACT: This preliminary study suggests that circulating microRNAs in the plasma may help serve as biomarkers of neural repair and aid in understanding human neural repair mechanisms. If validated in larger studies with appropriate controls, these markers could aid in timing rehabilitation therapy or designing recovery-based therapeutics.

ABSTRACT IMPACT: Improve healthcare of patients with Cystic Fibrosis by reducing the time needed to generate results. OBJECTIVES/GOALS: We developed an automated framework capable of segmenting the lungs, extract the airways, and create a skeletonize map of the airways from CT scans of Cystic Fibrosis patients. As future expansion, the framework will be expanded to measure the airways diameters, detect the abnormal airways, and count the number of visible airways generations. METHODS/STUDY POPULATION: For this study, 35 CT scans from CF patients with different levels of severity were used to test the developed framework. The lungs segmentation was performed using an algorithm based on Gaussian Mixture Models for mild cases, and for severe cases a technique that uses convex hull and the recurrent addition of ‘dots’ was implemented. The airways extraction was performed using a 26-points connected components algorithm in conjunction with a curve fitting technique over the histogram of voxel values. Medial axis transform was used to perform the skeletonization of the extracted airways, and airways diameters determined via ray-casting. RESULTS/ANTICIPATED RESULTS: The framework was able to correctly obtain the segmented lungs in all 35 sample volumes regardless of disease severity. In contrast, it tends to fail to skeletonize the airways for severe cases where the framework is unable to differentiate between abnormal lungs conditions and dilated airways. Fine tuning is required to achieve better results. The expected result of the future implemented sections of the framework are focused to characterize the extracted airways by: 1) measuring the airways diameters; 2) detect and count the number of abnormal airways sizes; and 3) count the number of visible airways branching which will permit determination of stage and grade of the lungs of CF patients. DISCUSSION/SIGNIFICANCE OF FINDINGS: The proposed framework allows a fast and reproducible way to segment the lungs and create a skeletonized map of the airways that are independent of clinical training. In addition, this framework will be extended to obtain measurements of airway dilation and branching level, which could provide a deeper insight of the airways in CF patients.

OBJECTIVES/GOALS: The goal of this study is to develop a multi-centered Translational Studio model that can help in the development of quality translational studies using resources from four different institutional partners (University of Texas Medical Branch, Texas Southern University, University of Houston Clear Lake and Houston Methodist). METHODS/STUDY POPULATION: We conducted two rounds of four Futures Workshops for a total participation of 28 stakeholders from four different partners. Future Workshops were used to critique, envision, and articulate novel “futures” that can be achieved at least partly through design practices (Muller, 2002). In the first round of workshops, we asked participants about their institutions’ strengths, weaknesses, resources and investigator needs regarding the Studio. In the second round we asked about different studio models, pros and cons of each model and guiding principles for a studio. Alongside a pragmatic content analysis, multi-stage deductive and inductive qualitative analyses were used to understand people’s views on the future of a multi-institutional Clinical Trials Studio. RESULTS/ANTICIPATED RESULTS: The first-round workshops’ analysis described peoples’ goals for what the studio should be. The future desired studio was described as guide, matchmaker, initiator and advocate. The second-round workshops’ analysis discussed the pros and cons of a variety of possible models including, centralized, decentralized, and topic-specific (and allowed other suggestions) while also describing principles for the guidance of a studio. Here the analysis showed people wanted certain characteristics for the studio (i.e. effective, efficient, locally-responsive, consistent, etc.). They also prescribed four principles that a studio should be guided by: non-hierarchical partnership, user-centeredness, respect/collegiality, and sharing. DISCUSSION/SIGNIFICANCE: The future workshops were useful in developing a shared multi-institutional Clinical Trials Studio model that is planned to be deployed in 2025. Participants valued a studio that was both directly supportive to participants and played a role in creating or advocating for institutional resources and policy for research.

OBJECTIVES/SPECIFIC AIMS: Metachromatic leukodystrophy (MLD) is a rare, lysosomal storage disorder caused by decreased enzymatic activity of arylsulfatase A. This can be the result of mutations in the ASA gene, or in rare cases PSAP. Historically, MLD has been subdivided into 3 forms based on age of onset: late infantile, juvenile, and adult. These subtypes were defined decades ago, prior to the appreciation of the full clinical spectrum of this lysosomal storage disorder and the advent of genetic testing. As a consequence, these empiric age-based historical definitions do not fully account for the spectrum of disease and are not founded in evidence-based analysis of phenotypic cohorts. Additionally, the antiquated definitions do not fully predict presenting features or disease course, and they fail to stratify outcomes in the few therapies currently available to treat this disease. As novel targeted therapeutics are developed, it is essential to have a clear understanding of the clinical presentation and natural history of MLD. Without properly defined sub-populations, it is difficult to design a therapeutic clinical trial that can demonstrate efficacy in a heterogeneous group. METHODS/STUDY POPULATION: In this project, we collected the retrospective natural history of over 50 individuals from around the world. We created an electronic database in REDCap to able to longitudinally collect clinical information. Using this retrospective natural history approach to understanding the disease course of individuals affected by MLD, we were able to characterize age of onset, delay to diagnosis, and common presenting features. RESULTS/ANTICIPATED RESULTS: Our results suggest distinct clinical phenotypic subgroups, with distinct presentations. DISCUSSION/SIGNIFICANCE OF IMPACT: With a better understanding of the natural history of MLD, we will be able to better counsel families and to design clinical trials with more coherent cohorts and more appropriate clinical endpoints.

Objectives/Goals: The expanding emphasis on translational science necessitates a rethinking of traditional academic formats. To align with the central themes of CTS, we have redesigned our PhD journal club and WIP sessions, introducing novel and innovative approaches that enhance the relevance of these activities to real-world scientific and clinical challenges. Methods/Study Population: The newly adapted journal club format for CTS Predoctoral students at Mayo Clinic maintains the traditional focus on literature review but now incorporates a structured analysis of the clinical implications and potential applications of the research. This innovation aims to foster a deeper understanding of how basic research findings can be translated into improved patient outcomes and healthcare practices. Similarly, the WIP sessions have been restructured to offer an engaging and dynamic learning environment designed to empower clinical and translational science predoctoral students to effectively present their research while emphasizing the challenges they have overcome, demonstrating the translational potential of their findings, and enhancing their communication skills. Results/Anticipated Results: Feedback from participants demonstrates strong support for the new format. Students report a greater engagement with the material and a clearer understanding of how their research can contribute to improving patient outcomes. Discussion/Significance of Impact: These changes accommodate the diverse projects in CTS and embody a commitment to pushing the boundaries of knowledge in CTS. This dual transition marks a significant advancement in preparing PhD students for careers in translational science, ensuring that their research is not only rigorous but also impactful in the real world.

Objectives/Goals: Most studies house mice at temperatures known to trigger moderate cold-induced hypermetabolism in addition to failing to consider sex as a biological variable, thereby limiting translatability. The major goal of this study was to evaluate metabolic responses to burn injury in male and female mice housed at standard and thermoneutral temperatures. Methods/Study Population: Following randomization, young male and female C57BL/6J mice (n=64; 16 per group, 50% male) were housed at room temperature (24°C; RT) or thermoneutrality (30°C; TN) for 14 days post sham or burn injury. Burn injuries involved 6.5cm2 in female mice and 8.6cm2 in male mice, translating to roughly 12.5% of the total body surface area. Energy expenditure was assessed through metabolic phenotyping, and DEXA scans (prior to injury and euthanasia) were performed to assess body composition. Semi-quantitative untargeted metabolomics were carried out on brown adipose tissue, heart, kidney, liver, plasma, spleen, and white adipose tissue. Results/Anticipated Results: Compared to TN, housing mice at RT resulted in 83% and 94% greater basal energy expenditure in male and female mice, respectively (p<0.0001 for both). Burns resulted in moderate hypermetabolism in mice housed at TN only, evidenced by ~20% increase in BEE (p = 0.037). Cachexia was greater in TN-housed mice, where males subjected to burn injury demonstrated greater BAT and WAT wasting when compared to RT housed mice (p < 0.01). At the proteomic level, RT housing masked the burn-stress response. Ingenuity pathway analysis and mitochondria-specific analyses revealed changes in the mitochondrial proteome were more pronounced in mice housed at TN compared to RT housed mice. Further, sexual dimorphisms in the global proteomic stress response to burn injury were more evident in mice housed at TN. Discussion/Significance of Impact: Housing mice below TN masks key physiological and tissue-specific proteomic responses to burn injury, thereby limiting the translational value of these models. Humanizing rodent physiology by housing mice at TN temperatures represents a tool to enhance the utility of mouse models to recapitulate stress response to burn trauma.

OBJECTIVES/GOALS: Recent studies have assessed the association between surgery with general anesthesia and cognitive decline in longitudinal cohorts of older adults. Patients diagnosed with dementia more frequently drop out of these longitudinal studies or are unable to complete the test battery. We revisit this aim with focus on methods for informative dropout. METHODS/STUDY POPULATION: We use data from the Mayo Clinic Study of Aging (MCSA), a longitudinal epidemiological study of the prevalence, incidence, and risk factors for mild cognitive impairment (MCI) and dementia. Our primary outcome of interest was global cognitive z-score, assessed at study visits every 15 months. We implement linear mixed effects models to assess the association between post-enrollment exposure to surgery/anesthesia and subsequent cognitive decline trajectories. Demented patients more frequently drop out of MCSA, so, subjects with the worst cognitive outcomes are unobserved and missing data may be informative. Since this missingness may be missing not at random, we use shared parameter models to analyze continuous cognitive outcomes while jointly modeling time to dementia. RESULTS/ANTICIPATED RESULTS: A total 1948 subjects, non-demented at baseline, from the MCSA were included. Median age was 79, 51% of subjects were male, and 16% had MCI at enrollment. Among median follow-up of 4 study visits over median 5.4 years, 172 patients developed dementia and dropped out from further assessments of cognitive function. In adjusted linear mixed effects models, our data suggest post-enrollment exposure to surgery/anesthesia is associated with a decline in cognitive function over time (change in slope = −0.07 standard deviations of cognitive z-score per year, 95%CI = −0.08, −0.05, p<.001). After adjusting for informative dropout using shared parameter models, surgery/anesthesia is associated with greater cognitive decline (change in slope = −0.14 per year, 95%CI = −0.16, −0.12, p<.001). DISCUSSION/SIGNIFICANCE OF IMPACT: We revisited a prior analysis by our group with consideration of informative dropout. Subjects who dropout due to dementia may have different trajectories of cognitive decline compared to non-demented subjects. Shared parameter models estimate the association between surgery/anesthesia and cognitive decline accounting for informative dropout.

OBJECTIVES/GOALS: To determine if decellularized costal cartilage (DCC), which could theoretically be obtained “off the shelf,” would provide similar results to autologous cartilage grafts previously studied in this lab, thereby widening the application of our novel nipple engineering approach to all patients undergoing nipple reconstruction. METHODS/STUDY POPULATION: PLA scaffolds (diameter: 1.0 cm, height: 1.0 cm) were printed using a PRUSA 3D printer and sterilized. Lamb costal cartilage was minced (1 mm3) or zested (<0.2 mm3) and then decellularized. The quality of decellularization was assessed using DNA quantification and histological analysis. DCC was then packed into PLA scaffolds and implanted subcutaneously into immunocompetent Sprague Dawley rats using a CV flap technique. The constructs were explanted and evaluated up to 6 months after implantation. RESULTS/ANTICIPATED RESULTS: All nipple reconstructions showed well-preserved diameter and projection due to persistence of the external scaffolds at 1, 3, and 6 months. Mass and volume of engineered tissue was well-preserved over all timepoints. Compared to implantation values, engineered zested nipples demonstrated a 12% mass increase and a 22% volume increase at 6 months. Minced nipples illustrated a similar mass and volume gain with a 21% increase in mass and a 13% increase in volume at 6 months secondary to infiltration of fibrovascular tissue and growth through scaffold wall pores, respectively. Histologic analysis demonstrated a mild inflammatory infiltrate 1 month after implantation which was replaced by fibrovascular tissue by 3 months that remained stable through 6 months. The processed DCC structure remained unchanged over time. DISCUSSION/SIGNIFICANCE: Using acellular ovine xenograft within bioabsorbable scaffolds, we have engineered neonipples that maintain their volume for at least 6 months. DCC architecture is well-preserved with minimal evidence of immune-mediated degradation. By using DCC, this novel approach to nipple engineering may be applied to any patient requiring reconstruction.

OBJECTIVES/GOALS: As hospitals across the nation respond to the need to address community violence, there is a dearth of Hospital-based Violence Intervention Programs (HVIPs) in the South despite having disproportionate rates. This research aims to identify key factors and strategies for implementation of an HVIP among rural patient populations in a southern state. METHODS/STUDY POPULATION: Semi-structured interviews will be conducted with medical providers, social service organizations, and patients transferred from four high-risk rural areas in Arkansas. Data will be analyzed using Framework Analysis, a rapid analysis approach involving framework development, code application, impactful statement identification, and content analysis. Evidence- Based Quality Improvement (EBQI), a group consensus making process, will be conducted to identify key implementation strategies and factors to adapt based interview findings. Priority areas for adaptation will be identified via systematic rating. The EBQI team, including researchers and key rural stakeholders will engage in a series of discussion, vote on final strategies, and develop a guide for future HVIP implementation and pilot testing. RESULTS/ANTICIPATED RESULTS: Findings from this study will result in a prioritized list of barriers and facilitators across sample groups. Factors will be rated by level of importance. Cluster maps will display the relationships among factors. Go and no-go zones will be identified based on importance and feasibility. Implementation strategies will be mapped to barriers and facilitators. DISCUSSION/SIGNIFICANCE: The findings will result in a culturally and geographically relevant HVIP model and package of implementation strategies to test in future hybrid trials (feasibility pilot & multi-site RCT); and shape the future of violence prevention efforts in healthcare settings across the rural South.

OBJECTIVES/GOALS: Febrile infants are a model population for exploring communication, implicit bias, and health disparities in the pediatric emergency department. Using mixed methods, we performed an in-depth analysis of disparities and physician-parent communication, a potentially modifiable driver of inequities. METHODS/STUDY POPULATION: We performed a multicenter cross-sectional study of febrile infants with low-risk for invasive bacterial infection evaluated between 2018-19 across 32 institutions. We performed logistic regression to assess the association between 1) race and ethnicity (Non-Hispanic White [NHW], Non-Hispanic Black [NHB], Hispanic [H], Other Race and/or Ethnicity [O]) and 2) limited English proficiency (LEP) with the standard of care, discharge without lumbar puncture and without antibiotics. We concurrently performed in-depth semi-structured interviews with physicians to explore physician–parent communication and implicit bias in the management decisions for febrile infants. RESULTS/ANTICIPATED RESULTS: Of 3629 low-risk febrile infants, 74.2% received standard of care. NHB, compared to NHW, infants were more likely to receive standard of care (aOR 1.3 [95% CI 1.0-1.7]; aOR H: 1.1 [95% CI 0.9-1.4]; aOR O: 1.0 [95%CI 0.8-1.4]). There was no association between LEP and standard of care (aOR 0.8 [0.61–1.1]). When accounting for documented use of professional interpretation, families with LEP without documented professional interpretation were less likely than families without LEP to receive standard of care (aOR 0.6 [95% CI 0.4–0.9]). Thematic analysis of 13 physician interviews is expected to elucidate aspects of the physician-parent interaction that influence management decisions. DISCUSSION/SIGNIFICANCE: Infant race, ethnicity, and use of professional interpretation, are associated with the management of low-risk febrile infants. Physician behaviors that are influenced by the physician-parent interaction may represent modifiable targets for future interventions to promote health equity.

Objectives/Goals: This poster details the development, implementation, and assessment of a comprehensive competency-based curriculum crosswalk and training plan aimed at enhancing the mentorship and skill development of translational research mentees within the National Institutes of Health (NIH) KL2 and TL1 Career Development Award programs. Methods/Study Population: The Center for the Improvement of Mentored Experiences in Research (CIMER) Mentoring Up for Early Career Investigators program, first developed by the University of Wisconsin-Madison, was further adapted by the Frontiers Clinical and Translational Science Institute (Frontiers CTSI). A competency crosswalk illustrates connections between a training curriculum and expected competencies. Developing a competency-based training crosswalk is a strategic approach designed to align mentoring practices with established NIH competencies, including the Seven Characteristics of a Translational Scientist, Mentoring Competency Assessment, Responsible Conduct of Research, Translational Teams, and TeamMAPPS. Results/Anticipated Results: The KL2 and TL1 Award Programs serve as ideal platforms for applying the developed curriculum. Implementing a competency-based, evidence-based, and culturally responsive curriculum for research mentee training has shown substantial benefits. Our pilot tests and full-scale implementation within the KL2 and TL1 Award Programs have demonstrated marked improvements in mentee competencies, such as technical skills, research design, and professional development as shown through evaluation feedback. Integrating the new Frontiers CTSI curriculum has fostered more effective and supportive mentoring relationships. Mentees have reported high satisfaction levels with the training program, particularly appreciating the interactive didactics, continuous feedback mechanisms, and reflective practices. Discussion/Significance of Impact: By adopting these recommendations, Clinical and Translational Science Awards (CTSA) and similar programs can improve research mentorship quality and impact, fostering a diverse cohort of skilled researchers. Implementing these strategies in CTSA mentorship programs offers a model for broader application in research training.

Objectives/Goals: To evaluate longitudinal trends in HPV vaccination initiation and series completion among adolescents aged 9–17 years in Puerto Rico from 2017 to 2024 and to determine the extent to which sustained coverage patterns following the 2018 school-entry mandate align with Healthy People 2030 targets. Methods/Study Population: A retrospective cross-sectional analysis was conducted using longitudinal registry data from PREIRS, Puerto Rico’s modernized immunization system integrating legacy PRIR records with electronic reporting. Records included adolescents with at least one vaccination event between 2017 and 2024 (N = 3,584,516). Vaccination status was categorized as initiated (≥ 1 HPV dose) or series completed (2–3 doses per ACIP recommendations) for each calendar year. Descriptive crosstabs and chi-square tests assessed annual differences (p < .001). Multiyear comparisons were performed independently for each cohort year to evaluate temporal changes, and stratified analyses were used to assess trends across sex and age groups. Data were analyzed using IBM SPSS Statistics (IBM Corp., Armonk, NY). Results/Anticipated Results: HPV vaccine initiation increased from 25.8% (2017) to 35.7% (2024), and series completion rose from 16.4% to 28.1% (χ²=18,498.85; df=7; p<.001). The steepest rise occurred between 2019–2021, coinciding with the school-entry mandate, followed by stabilization post-2021. Female coverage remained higher but differences narrowed annually. Publicly insured adolescents comprised over 80% of vaccine recipients, indicating socioeconomic influence. Coverage was highest among 11- to 13-year-olds, reflecting effective policy reach. Despite gains, both initiation and completion remain below the ≥80% Healthy People 2030 target, signaling the need for renewed HPV vaccination promotion and policy reinforcement among adolescents. Discussion/Significance of Impact: HPV vaccination gains in Puerto Rico demonstrate the sustained impact of the 2018 school-entry mandate and PREIRS infrastructure. Narrowing sex-based gaps indicate improved program consistency, while post-2021 plateau and public-insurance dominance highlight persistent socioeconomic barriers to long-term sustainability.