Objectives/Goals: Mayo Clinic’s Clinical Trials Beyond Walls™ (CTBW) program collaborates with study teams to implement decentralized elements in clinical trials, enabling participation from home or local settings. In cancer treatment trials, traditional paper pill diaries are replaced with real-time digital tracking solutions to monitor chemotherapy adherence. Methods/Study Population: The CTBW team developed a solution to deliver electronic pill diaries to research participants using the electronic health record (EHR) system Epic and patient portal MyChart1. The solution includes a portal message to remind participants to take chemotherapy. Medication dose, date taken, and reasons for missed doses (e.g., “I forgot” or “side effects were bothersome”) are captured. An automated in-basket notification system alerts the study team when predefined conditions are met. Configurable medication schedules ensure diaries are sent according to the prescribed frequency. Reports were generated to allow study teams to monitor all participant diaries. Results/Anticipated Results: The CTBW team implemented this digital pill diary in neuro-oncology trial NCT066250472. We anticipate the pill diary in the patient portal will enhance chemotherapy adherence by capturing real-time data in a platform widely used by Mayo Clinic patients. This patient-reported data is stored in the EHR, where it is accessible to providers and study teams. This allows for continuous monitoring, which facilitates a streamlined review of potential adverse events, improved compliance visibility, and timely treatment adjustments compared to paper-based or external solutions. The system also streamlines data entry, reducing human error and eliminating manual transcription. The created language and workflow templates allow the CTBW to scale this approach to future cancer trials Discussion/Significance of Impact: Decentralized clinical trial participants may never visit Mayo Clinic, making digital recording essential. The EHR-based digital pill diary enables continuous monitoring within a familiar system for providers and patients, increasing study team visibility, and allowing for earlier intervention in cases of non-compliance or adverse events.

Objectives/Goals: The research aims to prototype a mobile app for physicians to remotely monitor patients receiving CD19-directed CAR T-cell therapy post-discharge. This app will facilitate standardized data collection across carious CAR-T treatment centers and help harmonize follow-up protocols. Methods/Study Population: A literature review and semi-structured interviews with patients, clinical coordinators, and experts helped identify essential parameters for a mobile app prototype aimed at monitoring adverse effects such as cytokine release syndrome and neurotoxicity. The app was designed through process mapping to combine data from self-reports and wearable devices, such as the Garmin smartwatch. New screens were designed in Figma, drawing from an existing patient monitoring app for allogeneic stem cell transplant follow-up. Finally, a preliminary feasibility study will be conducted to gather feedback on the app prototype from CAR T-cell therapy patients, healthcare providers, and stakeholders, ensuring its effectiveness and usability. Results/Anticipated Results: Semi-structured interviews with people with professional and lived experience with CAR T-cell therapy were conducted to determine what metrics might be monitored and how they could be measured remotely to effectively monitor for side effects. The mobile phone application was then prototyped using process mapping in Visio®, designed in Figma, and preliminary development was completed. The final prototype includes parameters that will be recorded or measured using a combination of self-reporting and devices to monitor body temperature, basic vitals, activity, sleep, and cognitive function, among others. The prototype of the remote monitoring app is the first step in implementing remote monitoring of CAR T patients, standardized data collection, and reduction in the overall cost of CAR T-cell therapy. Discussion/Significance of Impact: This app will enable physicians to monitor patients for routine follow-ups and adverse effects, such as CRS and ICANS. Future research will validate the digitized ICANS assessment and used to establish best practices for standardizing CAR-T follow-up protocols across Canada.

Objectives/Goals: To determine the safety and feasibility of single-pulse transcranial magnetic stimulation (spTMS) for assessing corticospinal tract (CST) excitability and integrity in infants with perinatal brain injury, bridging foundational neuroscience to potential early diagnosis and clinical interventions during critical neuroplasticity periods. Methods/Study Population: Nineteen infants with perinatal brain injury underwent 1–3 spTMS sessions at three developmental time points: 3–6 months, 12 ± 1 month, and 18 ± 1 month. spTMS targeted the primary motor cortex to elicit motor-evoked potentials (MEPs), recorded via electromyography (EMG) from bilateral wrist flexor muscles. Safety monitoring included heart rate (HR), respiratory rate (RR), the Modified Behavioral Pain Scale (MBPS), and caregiver feedback. Feasibility was evaluated based on the ability to elicit MEPs, the number of trials that elicited MEPs, and procedure tolerability. Pre- and post-spTMS physiological and behavioral data were analyzed using linear mixed-effects models (LMEM) to account for repeated measures within subjects. Results/Anticipated Results: Thirty-five spTMS sessions were conducted in 19 infants (mean age 8.75 ± 5.12 months) with perinatal brain injury, delivering 1936 pulses with a median inter-pulse interval of 24.7 seconds. Analysis with LMEM found no significant changes in HR (mean difference  =  0.51 bpm, p  =  0.81) or RR (mean difference  =  0.69 breaths/min, p  =  0.66). MBPS scores showed a small statistically significant increase (mean difference  =  0.57, p  =  0.046), but overall remained low (mean score change from 1.94 to 2.51 on 0–10 scale). The median change score was 0, and 18/35 sessions showed no change in MBPS, indicating low discomfort with TMS. No adverse events were reported during or after the sessions. The feasibility of eliciting MEPs in this population was confirmed, with 235 MEPs identified in 17/19 participants. Discussion/Significance of Impact: Understanding neurodevelopment after injury is crucial for early diagnosis and targeted rehabilitation. Our study demonstrates that spTMS is a safe, feasible tool for assessing motor pathways in infants with early brain injury, highlighting its potential for clinical translation in neurodevelopmental disorders, and offering a pathway to improved care.

Objectives/Goals: This literature review examines the current landscapes of plain language summaries (PLS), which are used to make research accessible to nonexpert audiences. It aims to identify gaps in their implementation by focusing on challenges related to consistency, accessibility, and quality across fields. Methods/Study Population: A systematic search was conducted using databases such as PubMed and Google Scholar, employing key search terms like “plain language summaries,” “scientific communication,” “health literacy,” “patient education,” “knowledge translation,” “accessibility in research,” “public engagement,” “lay-person,” and “lay summaries.” Literature from multiple sources (pharmaceutical companies/industry, nonprofit organizations, private–public partnerships, and government) was compared to assess the gaps in current PLS best practices. Results/Anticipated Results: Search results yielded 95 articles. Of those, 37 articles fit the criteria, highlighting critical gaps in PLS implementation for clinical research. Preliminary findings suggest a lack of standards and guidelines, as well as a need for more research on the effectiveness of PLS for improving knowledge transfer and patient engagement. Key limitations were identified for investigator-initiated trials (IITs). A best practice table, comparing recommendations from each group of sources, was developed with suggestions for writing effective PLS. While there is consensus on some principles (i.e., importance of simplicity), differences emerge regarding optimal length and the use of layperson glossaries and graphics. The table aims to serve as a guide for creating effective and standardized PLS across fields. Discussion/Significance of Impact: There are limited PLS best practice resources tailored for IITs. These findings could lead to more practical tools and a streamlined approach to enhance communication strategies for lay audiences. This would benefit trial participants and community members who rely on this information and bridge the gap between scientific communities and the public.

Objectives/Goals: Ventilator-associated pneumonia (VAP) is an infection caused by bacteria, viruses, or fungi during mechanical ventilation. We analyzed a cohort of COVID-19 patients admitted to the intensive care unit with respiratory failure with different VAP outcomes. We hypothesize that the multiomics data can help predict VAP development within this cohort. Methods/Study Population: We recruited participants from a cohort on a NYU IRB protocol (i22–00616), who had COVID19 respiratory failure, admitted to ICU, and required invasive mechanical ventilation (n = 245). We collected and analyzed research specimens (bronchoalveolar lavage [BAL, n = 213], tracheal aspirates [n = 246], background [n = 18]) and clinical cultures (sputum and BAL) for 245 participants. A panel of experts adjudicated VAP within the cohort, resulting in 92 VAP diagnoses. We annotated metatranscriptome (Illumina NovaSeq) using a Kraken/Bracken database, and KEGG for functional annotation of transcriptome data (Illumina HiSeq). We used edgeR (v.4.0.16) to analyze differential expression of metatranscriptome and transcriptome data. Results/Anticipated Results: We diagnosed VAP in n = 92 (38%) participants. We found significant differences in days of overall hospital stay (p Discussion/Significance of Impact: VAP is a serious complication of mechanical ventilation, and oral commensals alter the lung microbiome and host immunity. We identified a transcriptome-metatranscriptome signature that identifies those at VAP risk. VAP was associate with both pro- and anti-inflammatory gene expression resulting in increased risk for lower airway infection.

Objectives/Goals: The demands on MICHR’s Evaluation team are profuse and varied. Quarterly team meetings were used to keep track projects, identify new projects, and relay important new initiatives from MICHR leadership. The MICHR Translational Innovation team took on the task of assessing the Evaluation team’s processes to design better workflow and effectiveness. Methods/Study Population: The process included 5 stages, Empathize, Define, Ideate, Prototype and Test. Sixteen interviews were conducted with MICHR faculty and staff. Interviews were coded and summarized. Seventeen themes were mapped and distilled into 5 key insights. From the key insights, design principles were identified to guide a design session with Translational Innovation staff and Evaluation staff. New work processes were proposed, designed, and tested by both teams. The Evaluation team “test-drove” the prototype and iterative design sessions were conducted to determine which new elements were successful. The Evaluation team was positioned to begin utilizing the newly designed process at the beginning of MICHR’s new grant year. Results/Anticipated Results: The MICHR Evaluation team is instrumental to the development, conduct, and dissemination of Clinical & Translational Science (CTS), a primary objective of MICHR’s work. Three types of evaluation projects were identified through the design process: required reporting, CQI/program improvement, and CTS/impact evaluation. The service design process enabled the Evaluation team, and MICHR program leads to better identify and prioritize collaborations between the Evaluation and program teams that improved the quantity and quality of MICHR CTS outputs. Discussion/Significance of Impact: Generating CTS is critical to the missions of NCATS and MICHR. Thoughtfully designing processes that facilitate and increase CTS output that can be shared and duplicated across the consortium is invaluable.

Objectives/Goals: Early childhood obesity is a major concern for Latin American children in the U.S., with gut barrier dysfunction as a key risk factor. Diet plays a role in gut development, but few studies have focused on Latin American infants. Our objective is to identify culturally relevant introductory foods that promote in vitro gut barrier development and function. Methods/Study Population: Pooled human milk (2.5 mL) from 6-month postpartum Hispanic mothers was combined with fruit and vegetable baby food products (2.5 g) and subjected to a 3-phase in vitro digestion system that simulates oral, gastric, and intestinal digestion. Digesta products were then anaerobically fermented for 24-hours using human stool inoculum, centrifuged, and filter sterilized. Intestinal epithelial cells (Caco-2, ATCC) were grown to confluence on 0.4 μm polystyrene transwell inserts using a DMEM + 10% FBS medium and allowed to differentiate for 21-days. Highly differentiated monolayers were treated with a 1:4 dilution of fermenta with medium in triplicate. The cell experiment was conducted twice. Cell layer integrity was measured using transepithelial electrical resistance (TEER) 24- and 48-hours after treatment. Results/Anticipated Results: Dietary intake data from the What We Eat in America database indicated that the top 3 fruit and vegetable exposures for infants with Mexican or Hispanic ethnicity were banana, apple, and carrot. Commercial baby food purees of these fruits and vegetables, in addition to baby foods with blueberry and spinach (Natural for Baby, Gerber Products Company) were acquired for digestion and fermentation experiments. Caco-2 cell experiments with these foods are ongoing. We expect Caco-2 monolayer incubated with fermenta from human milk and fruit or vegetables will have greater TEER values due to increased integrity of the cell layer as compared to those with breast milk alone. We also expect that exposure to fruit and vegetable fermenta will increase gene expression of tight junctions compared to exposure to media and human milk. Discussion/Significance of Impact: Using an in vitro digestion and fermentation system coupled with cell culture studies, we are identifying cellular mechanisms that link individual fruits and vegetables to gut barrier function. This will support translational work focused on mitigating obesity development in vulnerable populations.

Objectives/Goals: • Investigate culturally-specific beliefs, caregiving approaches, care preferences, and unmet needs among Asian American, Native Hawaiian, and Pacific Islander (AANHPI) care partners supporting persons living with dementia • Apply identified findings toward culturally-adapting caregiving resources for AANHPI dementia care partners Methods/Study Population: Qualitative data from the Better Together Dementia Care Study and Asian American, Native Hawaiian, and Pacific Islander (AANHPI) Dementia Caregiving Study were analyzed to investigate culturally-specific beliefs, caregiving approaches, preferences, and unmet needs of AANHPI dementia care partners. Both studies remotely conducted and recorded semi-structured interviews (1.5–2.5 hours), with care partners and providers though interview protocols were distinct. Transcripts were AI-generated, through Zoom or Trint, and analyzed using thematic content analysis by two coders. Apriori codes drawn from literature and inductively-identified codes were identified and coded. Preliminary findings informed sociocultural strategies used to adapt existing care partner resources for use in these groups. Results/Anticipated Results: Preliminary analysis of care partner (CP) interviews (N = 8; 4 romantic partners, 4 adult children), revealed differences in caregiving experiences and networks. CPs supporting care receivers (CR) who had emigrated away from extended family networks reported a lack of instrumental support. Most CR (7/8) had adult children, many of whom (6/8) provided some care, though half of CR-child relationships were distant or had past difficulties. Romantic partners were primary CPs while co-caregiving with children; Adult children served as primary CPs for unpartnered CRs (n = 4). Adapted CP resources integrated these findings, acknowledging the complexities of fulfilling traditional filial expectations in light of difficult past relationships. Formal services were framed as an extension of family-coordinated care. Discussion/Significance of Impact: Care partners of immigrants may have limited local family support and may benefit from formal services. Adult children may provide care, though this may be complicated by poor past relationship quality. AANHPI care partners may benefit from culturally-adapted resources which address these issues, though resource acceptability-testing is needed.

Objectives/Goals: Increased numbers of senescent cells have been detected in both traumatic brain injury and epilepsy, suggesting them as targets for therapeutic intervention for treating posttraumatic epilepsy (PTE) and underscoring the need for innovative methods to identify and target senescent cells as a means of alleviating pathology. Methods/Study Population: C57BL/6 mice will receive a single controlled cortical impact (CCI) before having their brains removed at 1 week, 2 weeks, 4 weeks, 1 month, 2 months, and 4 months post injury (n = 5 per time point). Brain sections will then be co-labelled for glial and senescent markers to observe which cells begin to express senescent markers at various time points. We will also perform single-cell RNA sequencing to observe genetic changes associated with both TBI and epileptogenesis. Mice will also be treated with navitoclax, a BCL2 inhibitor being investigated as a senolytic agent, to determine if treatment results in decreased senescence and epileptogenesis, as well as improved behavioral outcomes. Results/Anticipated Results: Preliminary data revealed that senescent microglia begin to arise in the mouse hippocampus as early as 1 week post injury and continue to increase in concentration over the course of the following month, with up to 25% of microglia expressing p16, a known marker of senescence. We anticipate that further staining will reveal senescent astrocytes and neurons in a similar time-dependent manner. Further, we hypothesize that the single-cell sequencing of microglia from injured mice will reveal alterations to the expression of genes associated with neuronal excitability, inflammation, and/or synaptic modeling, features known to be associated with epilepsy. Finally, we anticipate treatment with navitoclax will alleviate the senescent phenotype, resulting in decreased epileptogenesis and improved behavioral outcomes. Discussion/Significance of Impact: Considering the lack of any studies examining senescent cell prevalence in PTE, these data will be the first to identify these cells as etiological factors in PTE onset, as well as druggable targets for improving pathological outcomes in PTE patients.

Objectives/Goals: Children with intestinal failure rely on central venous catheters (CVCs) for at-home delivery of hydration and nutrition to sustain life. CVC associated infections are the leading driver of morbidity in this population. It is currently unknown what challenges caregivers of this patients face for at-home infection prevention tasks. Methods/Study Population: This is a qualitative analysis of the clinical implementation of at-home CVC care. Participants will be caregivers of children with intestinal failure in the Children’s Hospital of Los Angeles’ Intestinal Rehabilitation Program (a program with a large Spanish speaking population). The study team will conduct, record, and transcribe four synchronous remote focus groups using Zoom, two in English and two in Spanish, with 4–6 participants per focus group (n = 16–24). Questions will focus on caregiver experiences of CVC care. Transcripts will be analyzed using rapid qualitative analysis, an implementation science-oriented approach. The study team will review transcripts and summarize key points into matrices to examine relevant themes and quotes efficiently and systematically, using midpoint data analysis. Results/Anticipated Results: Illustrative quotes of all themes and domains discussed by caregivers will provide a person-centered overview of the specific types of facilitators and barriers to infection prevention task performance. Focus groups will be conducted in January 2025 with analysis in February. Facilitators and barriers will be mapped onto the Capability, Opportunity and Motivation behavioral model (COM-B), allowing for additional topics participants introduce by participants. Facilitators and barriers will affect all domains of the COM-B model and may also describe factors that cross multiple domains or fall outside the COM-B domains. Facilitators and barriers may differ between English and Spanish speaking caregivers. Discussion/Significance of Impact: Identifying facilitators of and barriers to infection prevention tasks will allow clinicians to maximize infection prevention efforts and improve quality of life for caregivers of children with intestinal failure. Additionally, this bilingual study will have improved external validity and address potential language-related barriers to care.

Objectives/Goals: 1. Examine structural, interpersonal, and health system factors that impact postpartum well-being for people who are racialized Black. 2. Differentiate components of postpartum well-being. 3. Design a model of postpartum care that addresses comprehensive well-being. Methods/Study Population: We conducted eight focus groups with participants in the Washington, D.C. area including four with Black birthing people who had given birth in the past two years (n = 23), and four with staff and providers from Community of Hope, a federally qualified health center, who provide care to birthing people (n = 19). We used an action-oriented qualitative approach informed by Black feminist theory. Our analysis was grounded in the 5D Cycle for Health Equity (reDefine, Discover, Dream, Design, and Deliver) and appreciative inquiry, which guide researchers to focus on strengths, be open to possibility, and engage radical imagination. Results/Anticipated Results: Participants reDefined postpartum health and wellness as physical, mental and social well-being, and material stability. Participants discovered that Black birthing people felt deeply unsupported navigating postpartum including difficulties with feeding, sleep, and mood and strongly believed that “postpartum” is at least a year, with different needs at different phases. Participants dreamed that postpartum care could be more accessible and trustworthy, have opportunities for social connection and creating a village, and have their basic needs (food, housing, clothing, and rest) met. Discussion/Significance of Impact: The participants’ conveyed that postpartum care must be designed and delivered to ensure that it is accessible, creates opportunities for connection, and promotes health, well-being, and joy. Postpartum care that can generate trust and engagement with healthcare, reduce morbidity and mortality, and increase thriving.

Objectives/Goals: As translational researchers, we seek to offer more treatments to more people, more quickly. Essential to this mission, we must demonstrate trustworthiness to establish trusted with those we seek to benefit. Research Jam uses a human-centered design (HCD) research approach to achieve this. Methods/Study Population: HCD is an iterative process that focuses on understanding people’s contexts while designing products and services collaboratively with the people who will interact with and be affected by the research. This generative approach helps participants express tacit and latent knowledge – emotions, needs, and desires that are often challenging to communicate verbally. We work with individuals who have lived experience relevant to the research being conducted and use HCD activities to help people share their thoughts, concerns, and ideas. These activities are highly interactive, promote collaboration, and explore topics with respect and sensitivity. Results/Anticipated Results: At Indiana Clinical and Translational Sciences Institute, we use the AAMC’s principles of trustworthiness. These principles parallel the tenets of HCD research; thus, taking an HCD research approach naturally builds trust between the researcher and participants and offers tangible benefit to the research, the researcher, and those with lived experience who participate. We have ten years of work collaborating with 40 PIs and over 600 participants. Our experiences demonstrate that this approach can create an environment where participants feel comfortable as they share their thoughts, concerns, and ideas which influences the research that could affect their lives. Discussion/Significance of Impact: An HCD approach helps demonstrate a researcher’s trustworthiness with the population being served. This is essential to achieve a lasting impact by discovering the best solutions from the community’s perspective and identifying partners to help implement solutions.

Objectives/Goals: Cerebral amyloid angiopathy (CAA) characterized by the accumulation of amyloid-beta in the cerebrovasculature, affects blood vessel integrity leading to brain hemorrhages and an accelerated cognitive decline in Alzheimer’s disease patients. In this study, we are conducting a genome-wide association study to identify genetic risk factors for CAA. Methods/Study Population: We genotyped 1253 additional AD cases using and curated existing genome-wide genotype data from 110 AD and 502 non-AD donors from the Mayo Clinic Brain Bank. We performed QC and imputation of all datasets. We conducted GWAS in AD only (N =  1,363), non-AD only, as well as the combined cohort (N = 1,865) by testing imputed variant dosages for association with square root transformed CAA using linear regression, adjusting for relevant covariates. To assess associations in the context of major CAA risk factors, we performed interaction analysis with APOEe4 presence and sex; and pursued stratified analyses. We collected peripheral gene expression measures using RNA isolated from 188 PAXgene tube samples of 95 donors collected across multiple time points. More than 1/3 of these participants have MRI measures collected. Results/Anticipated Results: Variants at the APOE locus were identified as the most significant in our study. In addition, several other variants with suggestive association were found under the main model adjusting for AD neuropathology (Braak and Thal). LINC-PINT splice variant remained associated with lower CAA scores in AD cases without the APOEe4 risk allele. To enhance the robustness of our findings, we are pursuing further expansion of our study cohort. In the periphery, we expect to identify expression changes associated with neuroimaging indicators of CAA and determine if variants and genes discovered via GWAS are implicated in these changes. Discussion/Significance of Impact: We expect this study will provide further insight into the genetic architecture underlying risk for CAA both in the context of significant AD pathology and without. Characterization of genetic variants and functional outcomes in the context of neuropathology may lead to new avenues of research aimed at identifying biomarkers and therapies to treat CAA

Objectives/Goals: Opioid use disorder (OUD) at delivery increased between 1999 and 2014. Clinical guidelines include medication for OUD (MOUD) for pregnant women with OUD and is associated with better fetal outcomes. Few large studies have compared prenatal MOUD outcomes to no MOUD. We evaluated the association of documented MOUD prescription during pregnancy with maternal outcomes. Methods/Study Population: We utilized aggregated electronic health records using the TriNetX platform to conduct a retrospective cohort study of females, aged 1249 years with a childbirth CPT code and documented opioid use via ICD-10 codes in the nine months before delivery between 2014 and 2020, comparing patients with MOUD prescription of buprenorphine or methadone during the nine months before delivery to demographically matched patients without any documented MOUD, using hazard ratios and 95% CIs for outcomes occurring one week to one or three years after childbirth. Results/Anticipated Results: MOUD cohort (n = 6,945, 85.33% White; 82.77% Non-Hispanic or Latino) was associated with significantly higher subsequent documented MOUD prescription (HR, 9.26 [95% CI, 7.98–10.76]; 6.21 [95% CI, 5.60–6.88]) and new remission codes (HR, 2.79 [95% CI, 2.15–3.62]; 2.85 [95% CI, 2.38–3.40]) at one and three years, lower ED visits at one year (HR, 0.88 [95% CI, 0.81–0.96]), no significant association of ED visits at three years (0.95 [95% CI, 0.89–1.02]), higher outpatient visits (HR, 1.26 [95% CI, 1.20–1.32]; HR, 1.27 [95% CI, 1.21–1.33], and no significant association of inpatient visits at one and three years (HR, 0.93 [95% CI, 0.813–1.06]; 1.06 [95% CI, 0.96–1.18]) than the never-MOUD cohort (n = 4,708, 76.11% White; 75.68% non-Hispanic or Latino). Discussion/Significance of Impact: A documented prescription for MOUD during pregnancy is associated with newly documented remission of OUD, increased outpatient visits, decreased ED visits, and additional documented MOUD prescriptions suggestive of increased access to continuity care. Efforts to increase MOUD use in pregnancy may improve maternal outcomes.

Objectives/Goals: As a priority area in translational science, rural health research can benefit from informatics methods for conducting thematic and environment scans. This study demonstrates an efficient approach to gaining insights about the rural health research literature by automated bibliometrics analysis. Methods/Study Population: We developed an automated pipeline to retrieve the 1972–2023 PubMed publications indexed with the MeSH terms “Rural Health” and “United States”. The article metadata in XML format were downloaded and parsed, including title, year, journal, author institutions, and MeSH terms. Each institution address was augmented by Google Maps API to obtain the county and latitude/longitude coordinates. Summary statistics were computed for the publication years, journals, author departments, and locations. A topic network was generated from the frequent co-occurring MeSH terms. The institutions were linked to Rural-Urban Continuum Codes and labeled on a map to visualize their geographic distribution. Results/Anticipated Results: A total of 4564 articles on rural health were analyzed. Two salient peaks of publications were revealed, one around 1978 and the other around 1993. The top author departments include Family Medicine, Nursing, Pediatrics, and Epidemiology. The five leading institutions reside in Chapel Hill, Minneapolis, Iowa City, Seattle, and Atlanta. The geographic distribution shows few institutions that reside in deep rural areas are well published on rural health, although the most scholarly productive institutions do seem adjacent to some moderately rural pockets. The frequently identified topics pertain to age group, study design, and specific concepts such as health services accessibility. Discussion/Significance of Impact: The two publication peaks in history were likely linked to certain policy milestone or seminal publication. Primary care and epidemiology departments have been most active in rural health research. Of concern, the geographic distribution of authors suggests under-investment in rural institutions.

Objectives/Goals: Explore and compare the functional mechanisms of song-based exercises compared to speech-language pathology exercises for dysphagia. The long-term goal is to increase patient outcomes through song-based programs that are accessible, enjoyable, and personalizable. Methods/Study Population: We will pilot the use of combined electroencephalography (EEG) and electromyography (EMG) technologies to analyze both central and peripheral contributors to laryngeal control in a cohort of healthy individuals. This approach provides detailed insight into the coordination between neural and muscular activity, which will serve as a baseline for future studies in clinical populations. Song-based vocal exercises will be compared with standard dysphagia exercises prescribed by speech-language pathologists to assess their mechanistic differences. Results/Anticipated Results: We anticipate identifying specific song-based tasks, such as variations in pitch, rhythm, and intensity, which differentially impact laryngeal musculature. Additionally, we will localize neural activation hotspots using EEG during these tasks, providing a more comprehensive understanding of how song-based therapy influences both peripheral and central mechanisms. Discussion/Significance of Impact: This project will lay the groundwork for developing evidence-based song-based therapies for dysphagia, providing an alternative to traditional SLP exercises. By creating an engaging therapeutic program, we aim to reduce dysphagia’s healthcare burden, including aspiration events, healthcare costs, and related mortality.

Objectives/Goals: Our objectives were to describe the characteristics of patients initially listed with inactive status on the liver transplant waitlist to identify if disparities exist and compare the prevalence of initial inactive status listings across transplant centers. Methods/Study Population: This was a retrospective cohort study of candidates waitlisted for liver transplant between March 1, 2023 and February 12, 2023, utilizing the Scientific Registry of Transplant Recipients, a national database including all solid organ transplant candidates. 224,736 candidates were included in analysis, and covariates included race, ethnicity, sex, age, body mass index, primary payer, MELD at listing, and etiology of liver disease. Results/Anticipated Results: Totally, 8,131 (3.62%) candidates were initially listed for liver transplant with inactive status. Although there were statistically significant differences between those listed initially with active status and those listed initially with inactive status in each covariate, these differences did not reach clinical significance. Of the 151 transplant centers, 128 listed any patients with an initial inactive status, with inactive status listings compromising 0–49.36% of total listings by transplant center. There is significant variation between listing centers in the practice of listing with initial inactive status by both liver disease etiology and across different eras of liver allocation policies. Discussion/Significance of Impact: Despite no significant clinical difference in the characteristics of patients listed with initial inactive status, there is significant variation across transplant centers of the prevalence of listing with initial inactive status. Subsequent investigations will focus on understanding these differences in listing practices between centers.

Objectives/Goals: Syncope is a common diagnosis in observation medicine, and characterization of observation patients is often limited to single unit, single center, or single payer systems. TriNetX, a federated deidentified multicenter national public database, provides an opportunity to study these patients from across the USA. Methods/Study Population: This retrospective cohort study queried data from 56 health care organizations (HCO) in TriNetX to examine differences between observation patients with syncope who required admission vs. those who were discharged. All observation stays with a diagnosis of syncope, defined by ICD-10, CPD, and SNOWMED codes, were queried. A total of 281,162 observation encounters were included in analysis, of which 46.4% (n = 130,357) were admitted and 53.6% (n = 150,805) were discharged. Data on age, gender, race, ethnicity, presence of congestive heart failure, EKG, and serum labs were collected for comparative analysis. T-test and Chi-square analyses were deployed with significance  =  p Results/Anticipated Results: The cohorts demonstrated statistically significant differences across all demographic factors, however, they were not clinically meaningful. Clinically significant differences include that only 72.8% of admitted patients and 68.3% of discharged patients had an EKG recorded in TriNetX during the period of observation (p Discussion/Significance of Impact: Patients admitted from observation status were more likely to have CHF, higher BNP, and pro-BNP values. TriNetX is a powerful tool to study patients across multiple hospital systems and payer types. Limitations, however, include incomplete data and inaccuracies among claims records.

Objectives/Goals: Our research addresses critical gaps by examining the prevalence, blood concentrations, and health implications of xylazine abuse, with a focus on its cardiotoxic effects. We aim to map the geographic distribution of xylazine use across Puerto Rico and characterize its impact on cardiomyocytes at relevant exposure levels. Methods/Study Population: To accurately detect and quantify xylazine in blood samples, we will employ chromatographic techniques coupled with mass spectrometry (UPLC/MS or GC/MS). The xylazine prevalence across the island will be mapped based on health system classifications. Samples will be categorized according to the eight healthcare regions of Puerto Rico, as defined by the “Administración de Seguros de Salud” (ASES), ensuring comprehensive geographic representation. We will investigate the expression profiles of proteins associated with cardiac injury and dysfunction in human cardiomyocytes and in the blood of drug users. Blood samples will be provided by “Iniciativa Comunitaria.” We will assess the xylazine effects on human cardiomyocyte viability and identify key biomarkers of cardiotoxicity induced by xylazine exposure. Results/Anticipated Results: In previous research, we demonstrated that xylazine induces cell death in endothelial cells through both extrinsic and intrinsic pathways. We also observed an increase in reactive oxygen species (ROS) levels after drug exposure, indicating oxidative stress as a potential mechanism of toxicity. Additionally, DNA damage was detected. Given the known relationship between endothelial damage and cardiomyocyte dysfunction in drug-induced cardiotoxicity, we hypothesize that xylazine concentrations vary regionally within Puerto Rico and that chronic xylazine abuse will elevate markers of cardiac injury and dysfunction at common user doses. Discussion/Significance of Impact: The increasing xylazine abuse, particularly in Puerto Rico, represents a critical public health challenge. Our study will fill a knowledge gap by providing crucial data on xylazine’s cardiotoxicity and mapping its geographic prevalence, with the potential to advise healthcare approaches and improve care for drug-using Hispanic populations.

Objectives/Goals: Aged keratinocytes are less proliferative than adult, and aged skin heals more slowly. We examined the proliferation kinetics of aged and adult human keratinocytes. We then tested whether an extrinsic agent, sh-oligopeptide-72, can ameliorate these defects. Methods/Study Population: We used live cell imaging (LCI) to examine the proliferation kinetics of aged (73–92y) and adult (34–49y) passage zero human keratinocytes. We then incubated aged keratinocytes with a peptide, sh-oligopeptide-72 (purported to improve keratinocyte proliferation), or vehicle (PBS). Lineage trees of cell divisions were constructed to determine cell cycle duration and the proliferation/differentiation outcomes of each division. To assess wound healing, cells were isolated from 3 patients, 82–92y, and plated in 2-well culture dishes with inserts. Wells were treated with sh-oligopeptide-72 (100 ng/ml) or vehicle (PBS). At confluence, the insert was removed leaving a well-defined 500 μm gap. The time until 100% closure of the defect was obtained using LCI and the wound healing size tool. Results/Anticipated Results: There was no significant difference in the number of stem cell (SC) colonies between aged and adult keratinocytes. However, aged keratinocytes produced more aged committed progenitor (CP) colonies (P<0.0001). Adult CP, but not stem, colonies were significantly larger than aged (P = 0.0001), and this was associated with earlier terminal differentiation (P = 0.0005). Aged SC and CP colonies exhibited a higher proportion of differentiation divisions, and their cell cycle duration (CCD) was increased. Sh-oligopeptide-72 rescued the increased terminal differentiation as well as decreased the CCD in SC colonies. Sh-oligopeptide decreased the mean closure time of the wound assays (143h vs. 204h, P = 0.04). Discussion/Significance of Impact: Sh-oligopeptide-72 reversed many of the proliferation defects that develop in aged SC colonies. Wound assays show that this results in improved keratinocyte function. These results suggest that the age-related changes in growth dynamics can be modified in response to extrinsic signals in vitro.