Objectives/Goals: Team science (TS) competency is important for translational science team collaboration. However, there are few educators available to assist teams. Asynchronous learning is an effective strategy for delivering TS content. The goal of this project is to expand TS education by providing online access to our learners using online modules. Methods/Study Population: The Collaboration and Team Science (CaTS) team at the University of Cincinnati provides a robust TS education and training program. As the need for team science gains recognition, CaTS has received increased requests for services, leading to a need to broaden TS offerings. To address this demand, the CaTS team created “Team Science 101,” an online, asynchronous, series of 15 modules covering basic team science concepts. Each module consists of an educational recording lasting an average of 20 minutes, optional topic resources, pre- and post-module surveys assessing learners’ confidence and satisfaction, post-module knowledge checks, and evaluation questions. Upon completing all modules, participants receive a completion certificate. Results/Anticipated Results: TS 101 will be piloted with a group of participants who expressed interest in asynchronous TS content and will be adjusted based on the feedback received. The associated pre- and post-module survey, post-module knowledge check, and evaluation questions will be monitored to determine learning levels and improve TS 101 overall. Canvas is the educational platform that houses these modules, allowing for participant follow-up and scalable dissemination. The CaTS team plans to disseminate TS 101 nationally and internationally for anyone interested in this resource. Discussion/Significance of Impact: There is a national effort to collect and curate TS education, training, and toolkits. TS 101 will be a useful educational tool that will expand the reach of team science educators, provide the foundation for educators to explore topics more deeply by building on the module topics, and provide education to broader audiences who lack access to TS experts.

Objectives/Goals: Triple-negative breast cancer (TNBC) is a highly aggressive form of breast cancer (BC) with limited treatment options. Mortality rate is especially high in African American (AA) women of reproductive age. High levels of intracellular calcium (Ca2+) have been shown in TNBC cells. This study is to investigate Ca2+ channel blockers (CCBs) as therapy for TNBC. Methods/Study Population: Two human TNBC cell lines obtained from ATCC – HCC1806, and MDA-MB-453 are treated with CCBs, Cilnidipine (Cil), and Mibefradil (Mib), in a concentration- and time-dependent manner. Cell proliferation assays are performed by the MTS cell viability assay. Intracellular Ca2+ levels are measured using the fluorescent dye: Fluro 4-AM. Apoptosis is determined by flow cytometry using Annexin V staining and mitochondrial permeability will be assessed by the Mito JC-1 assay. Expression of Ca2+ signaling genes will be quantitated by real-time polymerase chain reaction (RT-PCR). Potential pathways of CCB efficacy will be identified by ingenuity pathway analysis (IPA). Results/Anticipated Results: Our findings show both CCBs decrease cell proliferation in a concentration- and time-dependent manner to a maximum of 80% vs. control in both TNBC cells. Flow cytometry findings on both TNBC cells treated with both drugs at 20 µM for 24 hours depicts late apoptosis. Interestingly, Mib did not change the intracellular Ca2+ level in HCC1806 cells yet decreased in MDA-MB-453 cells by fivefold, while Cil increased the intracellular Ca2+ level in both cells almost twofold. It is anticipated that Mito JC-1 assay depict decreased mitochondrial potential in both cells. For reverse transcription polymerase chain reaction, it is anticipated that CCB treatment will increase transient receptor potential Ca2+ channels and decrease voltage-gated Ca2+ channels in both cells. IPA analysis is expected to show apoptotic pathways are involved in TNBC via CCB treatment. Discussion/Significance of Impact: TNBC lacks the estrogen receptor, progesterone receptor, and human epidermal growth factor receptor 2. Treatment options for TNBC remain severely limited. Our findings that both Cil and Mib can inhibit proliferation of human breast cancer cell lines indicate repurposing CCBs as treatment for TNBC warrants further investigation.

Objectives/Goals: Large-scale tumor sequencing efforts have led to annotations of novel cancer hotspot mutations that may underlie driver or cooperative function. We have sought to define the molecular consequences of such hotspots associated with pediatric DICER1 syndrome cancers, with the ultimate goal of revealing novel targets that may inform new standards of care. Methods/Study Population: We have performed genomic analysis to identify tumor types (in TCGA and MSK-IMPACT patient data) for which mutations in the Dicer1 gene (encoding Dicer protein) emerge as the dominant signature of driver function. As Dicer is a critical RNA processing factor responsible for the generation of microRNAs, which are posttranscriptional gene regulatory molecules, we have modeled these mutations in human embryonic stem cells in order to study the direct effects on miRNAs and their target genes in an isogenic background. In addition to providing the required setting for unambiguous attribution of function to specific mutations, clonal human ES cells offer an opportunity for modeling of both developmental and cancer requirements associated with altered Dicer function. Results/Anticipated Results: Through generation of genomics and functional datasets from matched genotypes in Dicer mutated human ES cells, we have identified specific alterations in miRNAs and their effects on target genes. Unexpectedly, we found direct evidence for both loss of function and gain of function attributable to Dicer mutations. In addition, through integrated analysis of genomic data from tumor sequencing datasets and our human ES cell models, we have identified potential miRNA and target gene alterations that underlie tumorigenic potential, nominating gene candidates for targeted therapy in DICER1 syndrome. Direct mouse modeling of such candidate gene targets has revealed evidence for driver function of identified miRNA and their targets. Discussion/Significance of Impact: DICER1 syndrome cancers comprise a wide variety of rare pediatric tumor types. Presently, we still lack an effective standard of care. Furthermore, the previous lack of molecular profiling precluded targeted therapy opportunities. Our precise knock-in modeling of Dicer hotspots and deep profiling of relevant tumors now provide candidate targets.

Objectives/Goals: Youth with IBD have preventive, psychosocial, and acute care needs beyond those of peers, yet receipt of services does not match those needs. Our objectives are to assess the feasibility of (1) an individualized care plan intervention to improve perceived and measured care quality and (2) a pragmatic trial design embedded in pediatric IBD practice. Methods/Study Population: This is a pilot rollout-design randomized trial (n = 60) at a regional academic medical center. Eligible patients are 13–19 years old with IBD for at least 3 months and scheduled for a follow-up visit during the trial. Research staff recruits from one cluster at a time until goal enrollment (14–16). Enrollees are randomized 1:1 to intervention (MyIBD now) or control (MyIBD after the trial). MyIBD combines a tabular summary of individualized acute, chronic, and preventive care needs with nurse facilitator support for patients to use the information. Surveys at baseline, 6 and 12 months measure care quality (Patient Assessment of Chronic Illness Care scale, vaccines, health services) and patient self-management skills (Partners in Health scale). Implementation outcomes are collected via chart review. Results/Anticipated Results: To date, 44 subjects have been randomized. Among subjects, the mean age is 16 years; 73% have Crohn’s disease, 77% have commercial insurance, 75% receive anti-TNF therapy, and 14% live in a rural area. Mean baseline perceived care quality (PACIC scale) is 76.9 (sd 16.3; out of 100); mean baseline perceived self-management skill (PIH scale) is 78.1 (sd 13.4; out of 96). On objective care quality measures, 59% have completed the HPV vaccine series, 32% have received an additional pneumonia vaccine; in the past year 68% have had a screening for mood disorders, 20% an emergency department visit for IBD, and 18% an IBD hospitalization. To date, the IBD clinical team has achieved 100% completion (intervention subjects receive MyIBD plus nurse facilitation) and 0% contamination (control subjects inappropriately receive MyIBD). Discussion/Significance of Impact: Study results to date support the feasibility of the pragmatic, embedded trial design and indicate opportunities for improvement in care quality as perceived by patients and as measured by common preventive and acute care quality indicators. An individualized care plan supported with nurse facilitation may improve pediatric IBD care quality.

Objectives/Goals: Pediatric inguinal hernia (PIH) is a common diagnosis managed by pediatric surgeons. Patients typically present in the outpatient setting for evaluation. However, a subset present emergently with hernia incarceration, which has a higher risks. Our research aims to identify and address disparities in access to care of PIHR. Methods/Study Population: This study has a sequential multi-methods approach. First, we will perform a retrospective cohort study of pediatric patients (ages 0–18) who have undergone PIHR at St Louis Children’s Hospital (SLCH) over the past 5 years (n =  3,421). We will assess the impact of demographic and system-based factors on patient outcomes. Second, we will conduct interviews with caregivers of patients who have undergone PIHR. Qualitative interviews will be coded to prospectively identify themes in barriers and facilitators to accessing care, understanding of disease, and effectiveness of risk factor education by the pediatric care team. Finally, we will analyze the readability index (RI) of patient provided materials to assess the impact of health literacy on access to care and PIHR outcomes. Results/Anticipated Results: We hypothesize that demographic factors and socioeconomic status (SES) impact outcomes in PIHR. We anticipate higher rates of incarceration and emergent presentation of children from low SES and racial/ethnic racial minorities in our dataset, with longer time from initial presentation to definitive repair. Moreover, we anticipate identifying significant barriers to accessing care for non-white patients and patients with low SES, with gaps in understanding about the risk factors, presentation, and management of PIH. Finally, we hypothesize that the RI of our patient provided materials do not match community literacy rates, and there will be opportunities to implement new patient provided materials to improve parental education and communication in the perinatal space. Discussion/Significance of Impact: More than 600 patients undergo PIHR annually at a large tertiary care pediatric hospital, with those who are born premature being at higher risk. Using a mixed-methods approach, we aim to implement targeted interventions to facilitate access to care and improve patient outcomes in PIHR.

Objectives/Goals: A sample of 1,073 hypertensive adults in Bangladesh showed unusually high rates of hypokalemia. We analyzed sociodemographic and clinical factors – including age, sex, weight, body mass index, blood pressure, creatinine clearance, and urine protein concentration – to identify key predictors of hypokalemia. Methods/Study Population: A cross-sectional analysis was conducted on 1,073 hypertensive adults from the OK study in Dhaka, Bangladesh (2022–2023). Hypertension was defined as blood pressure >140/90 mmHg, and none of the participants were on any antihypertensive medication prior to the study. Hypokalemia was defined as serum potassium Results/Anticipated Results: The prevalence of hypokalemia was 21.5%. In univariate analysis, age (OR 0.975, 95% CI [0.959, 0.990], p  =  0.00189), systolic blood pressure (OR 1.02, 95% CI [1.00, 1.03], p  =  0.00568) and diastolic blood pressure (OR 1.03, 95% CI [1.01, 1.04], p  =  0.000272) were significantly associated with hypokalemia. In multivariate analysis, age (OR 0.961, 95% CI [0.936, 0.985], p  =  0.00167) remained significant, and creatinine clearance (OR 0.986, 95% CI [0.974, 0.999], p  =  0.0386) became significant. BMI, weight, systolic and diastolic blood pressure, sex, and urine protein concentration were not significant. Discussion/Significance of Impact: In the multivariate analysis, age and creatinine clearance was statistically significant; however, their effect sizes were small and unlikely to be clinically meaningful. The underlying reasons for the high prevalence of hypokalemia in this population remain unclear, warranting further investigation to identify contributing factors.

Objectives/Goals: Despite persistent health disparities, rural individuals are underrepresented in clinical trials, due in part to access barriers. We investigated if targeted strategies enhanced recruitment, engagement, and retention of rural adolescents in the TEENS+ randomized clinical trial, a 4-month family-based behavioral weight loss intervention. Methods/Study Population: Adolescents (12–16 y) and parents with obesity were eligible for TEENS+. Treatment converted to virtual in COVID-19, allowing eligibility to expand to more rural areas. We leveraged Informatics, a practice-based research network, and direct marketing to identify potential rural participants. Targeted engagement strategies included: rural physician outreach, physician-endorsed letters, providing tablets and mobile hotspots, reimbursing travel, and offering in-person or remote assessment visits. Chi-square tests evaluated differences in screener completion and enrollment of rural families before (T0) and after (T1) changes were made. Noninferiority tests evaluated rural vs. nonrural retention and engagement (% attendance, % dietary self-monitoring) and engagement based on digital tool receipt. Results/Anticipated Results: N = 211 dyads enrolled (n = 54 in T1: 48% male; 41% Black). Screener completion by rural families significantly increased from T0 (9.8%) to T1 (15.1%; p = .043). Yet, there was no significant change in rural adolescent enrollment (T0 = 10%; T1 = 9%; p = .844). Sixteen adolescents (30%) received study tablets, and none needed mobile hotspots. Mean adolescent attendance was 75%±28% for group and 94%±18% for individual sessions, with no significant differences based on rural status or tablet use. Rural adolescent self-monitoring (via app) was 28%, compared with 50% for non-rural adolescents (p = .074). Retention was 94% at 4m and 89% at 8m for T1 participants, with no differences based on rural status. At the primary endpoint (12 m), retention was significantly higher for rural (100%) vs. non-rural (87%) participants; p = .013. Discussion/Significance of Impact: Rural adolescent screener hits increased yet enrollment was unchanged. However, rural attendance was comparable and retention exceeded, compared to nonrural participants. Strategies to yield equitable representation and engagement in clinical trials are essential for geographic generalizability and to reduce rural health disparities.

Objectives/Goals: Cerebral amyloid angiopathy-related inflammation (CAA-ri) is a spontaneous inflammatory cerebral vasculopathy that mimics complications of Alzheimer’s disease immunotherapies. Our objective is to evaluate imaging and cerebrospinal fluid (CSF) markers of blood–brain barrier (BBB) impairment and inflammation in CAA-ri. Methods/Study Population: We plan to enroll 20 patients total: 1) 10 patients with CAA-ri as defined by Auriel et al (JAMA Neurology 2016) (exposure group). 2) 10 patients with non-inflammatory CAA defined using Boston criteria 2.0 that do not also meet criteria for CAA-ri (control group). The primary outcome will be Ktrans, a parameter of BBB impairment calculated from dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) of the brain. Secondary outcomes will include DCE-MRI parameter VL, CSF albumin index, CSF fibrinogen, CSF sPDGFR-β, CSF MMP-2, CSF MMP-9, CSF C3, CSF IL1β, CSF IL6, IL8, and TNFα. Statistical comparisons between the exposure and control groups will be made using Wilcoxon rank sum test. Results/Anticipated Results: We anticipate significantly higher levels of BBB impairment and inflammatory biomarkers from DCE-MRI and CSF in subjects with CAA-ri relative to control subjects with non-inflammatory CAA. Discussion/Significance of Impact: Biomarkers are essential to characterize risk factors, pathophysiology, and possible treatment targets in CAA-ri. We plan to use the results of the current study to inform longitudinal studies that will test whether these markers are useful in identifying not only the presence of CAA-ri but also severity, progression, and response to treatment.

Objectives/Goals: We designed a forum to educate participants about bioethical issues in the application of big data (BD) and artificial intelligence (AI) in clinical and translational research (CTR) in underrepresented populations. We sought to determine changes in participants’ interests in ethics, bias, and trustworthiness of AI and BD. Methods/Study Population: 141 individuals registered for the forum, which was advertised to our partner institutions, minority-serving institutions, and community organizations. Registrants received email instructions to complete an AI Trustworthiness (AI-Trust) survey, a questionnaire with integrated qualitative and quantitative measures designed to better understand learners who engaged with the institution-specific AI/Data Science curriculum. Respondents completed the survey using personal devices via a link and QR code, with anonymized responses and enhanced privacy features. 82 people attended; 22 responded to the survey pre-forum and 22 post-forum. Pre- and post-forum responses were qualitatively compared to assess shifts in attitudes toward AI and BD and related interests in ethics, bias, and trustworthiness. Results/Anticipated Results: We found increased interests post- vs. pre-forum in the use of AI for CTR, AI bias and its effects on underrepresented populations, and ethical risk assessment and mitigation strategies for the use of BD to empower research participants. In contrast, trust in AI was lower post- vs. pre-forum. Moreover, respondents also indicated that the current application of AI in healthcare practice would result in increased racial, economic, and gender bias. In comparison, interest in ethical challenges, bioethical considerations, and trustworthiness regarding use of BD and AI in health research and practice did not differ pre- vs. post-forum. Discussion/Significance of Impact: Interest in the application of BD/AI in CTR increased post-forum, but AI distrust and bias expectations also increased, suggesting that learners become more skeptical and discerning as they become more knowledgeable about the complexity of the ethics of AI and BD use in healthcare, especially its application to underrepresented populations.

Objectives/Goals: This study aims to harmonize quality indicators (QIs) across University of Toronto-affiliated microbiology labs to establish universal benchmarks that enhance performance, patient safety, and health outcomes. Harmonized QIs will enable effective comparisons and enhance the consistency of care. Methods/Study Population: The study employed the Delphi method, a structured and iterative process to build consensus. An expert panel of clinical microbiology trainees, medical microbiologists, trainees, and site leads from five University of Toronto-affiliated microbiology labs was assembled. Initial insights were gathered through surveys and a comprehensive scoping review of the literature. The study involved two rounds of feedback, a SurveyMonkey-based survey, with a defined consensus of 75% agreement among participants. Followed by an implementation survey conducted through REDCap to assess how these QIs were adopted in practice and identify barriers to implementation. Results/Anticipated Results: The study achieved consensus on nine high-impact quality indicators, including blood culture volume and contamination rates, cerebrospinal fluid transport time, and turnaround times for Gram stain results. Blood culture contamination and positivity rates garnered the highest agreement, at 100% and 91%, respectively. While some indicators were widely accepted and implemented, others faced resistance due to feasibility concerns. The study also identified significant variability in the level of adoption across the participating laboratories, pointing to operational challenges and the need for further efforts to address these barriers. Discussion/Significance of Impact: This study highlights the importance of QI harmonization in improving lab services and patient safety. It reveals challenges in standardizing practices but promotes uniformity in QIs, laying the groundwork for better inter-lab collaboration, consistent outcomes, and improvements in microbiology.

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/Goals: The study is designed to co-develop a genetics education digital platform with community participants. Diverse populations, especially African American communities, are less likely to participate in genetic testing and clinical trials. To increase participation and community input, we want community participants across Louisiana. Methods/Study Population: A cross-sectional, mixed-methods study will be conducted to assess interest in learning about genetics through a digital education platform and to adapt the platform based on participant feedback. Specific Aim 1 will be achieved by recruiting a diverse cohort and collecting demographic data to identify participants’ characteristics. For Specific Aim 2, 35 community participants will be enrolled as a co-design team to complete 5-week educational modules. Feedback from focus groups will guide iterative platform refinements, ensuring the platform is culturally tailored and user-friendly. Results/Anticipated Results: For Specific Aim 1, we anticipate that the majority of participants will express interest in learning about genetics through the digital platform, with demographic data revealing a diverse participant pool, predominantly from African American and Hispanic communities. For Specific Aim 2, we expect that all 35 co-design team members will complete the 5-week modules. Feedback from the focus groups is anticipated to highlight the need for more user-friendly navigation, culturally tailored content, and enhanced visuals. These insights will guide the refinement of the digital platform for improved engagement and accessibility. Discussion/Significance of Impact: This study addresses a critical translational barrier – underrepresentation of African descended communities in genetic testing. By developing a culturally tailored, digital platform to engage these populations, the project aims to reduce health disparities, enhance genetic literacy, and foster inclusivity in genomic research.

Objectives/Goals: We evaluated the implementation of a peer-facilitated research best practices training for Community Health Workers and Promotoras (CHW/Ps) at four new partner sites to increase the capacity and capability of a workforce increasingly involved in community-engaged research. Methods/Study Population: Staff were trained using a train-the-trainer model, and materials were disseminated to partners at three academic institutions and one community-based organization. Each site delivered the training virtually or in-person in English and/or Spanish. CHW/P learners at all sites completed online evaluation surveys about the impact of the training on their knowledge and skills for participating in research-related work, and two CHW/Ps from each site participated in follow-up interviews to gather feedback about their experiences. Staff completed fidelity monitoring, follow-up interviews, and three brief surveys regarding feasibility, acceptability, and appropriateness of implementing the training. Results/Anticipated Results: The four sites conducted six trainings with a total of 42 CHW/Ps. Two sites each conducted one in-person training in English while the other two sites each conducted two virtual trainings, one in English and one in Spanish. Staff noted facilitators to successful implementation, including providing a facilitator guide and course materials in both languages and tips sheets for navigating REDCap; using the train-the-trainer model; and compensating CHW/P learners for attendance. The primary barrier noted was not having a budget for in-person trainings (e.g., refreshments, printed materials). CHW/P learners reported positive experiences with few suggestions for improving the training. Discussion/Significance of Impact: Preliminary results suggest the research best practices training for CHW/Ps is feasible, acceptable, and appropriate for implementation by partners at academic institutions and community-based organizations, regardless of language (i.e., English or Spanish) or delivery (i.e., virtual or in-person).

Objectives/Goals: Sexual minority populations report a disproportionately high prevalence of alcohol use, often attributed to coping with bi/homonegativity and systemic inequities across various social domains. This study aims to explore alcohol use patterns and associated neighborhood and individual factors among sexual minority populations (SMPs) using data from the NIH All of US dataset. Methods/Study Population: Alcohol use was assessed using the AUDIT-C (Alcohol Use Disorders Identification Test—Consumption) scale across a sample of 9,454 gay, 15,284 bisexual, 5,267 lesbian, and 349,748 straight participants. The AUDIT-C measured hazardous alcohol use, and logistic regression models were employed to examine its association with neighborhood-level factors (e.g., socioeconomic status, alcohol outlet density) and individual-level factors (e.g., age, race/ethnicity, income, and education) among SMPs. Interaction terms assessed how these relationships varied by sexual orientation. Sensitivity analyses were conducted to assess the robustness of the findings, including stratified analyses by gender identity and the exclusion of extreme outliers in alcohol use reporting. Results/Anticipated Results: Our analyses revealed that gay participants had the highest AUDIT-C scores (mean  =  3.60, SD  =  2.27), followed by bisexual (mean  =  3.35, SD  =  2.21), other SMPs (mean  =  3.18, SD  =  2.19), lesbian (mean  =  3.04, SD  =  2.08), and straight individuals (mean  =  3.05, SD  =  2.06). Alcohol use was positively associated with neighborhood disorder (β  =  0.12, 95% CI  =  0.07, 0.17), housing insecurity (β  =  0.14, 95% CI  =  0.03, 0.25), and male gender (β  =  0.98, 95% CI  =  0.96, 1.00). In contrast, neighborhood density (β  =  -0.11, 95% CI  =  -0.15, -0.07), food insecurity (β  =  -0.14, 95% CI  =  -0.20, -0.08), being Black, and identifying as bisexual were negatively associated with alcohol use. Sensitivity analyses determined no significant differences among specfic supgroups. Discussion/Significance of Impact: This study highlights important differences in alcohol use across SMPs and emphasizes the influence of neighborhood-level stressors (e.g., disorder and housing insecurity). These findings underscore the need for addressing social and environmental determinants of alcohol use in SMPs to mitigate the negative impacts of alcohol consumption.

Objectives/Goals: As life expectancy increases in people with HIV, neurocognitive impairment is becoming more common, and women with HIV (WWH) are disproportionately impacted. This work investigated mitochondrial function and oxidative stress in WWH in order to understand the relationship between mitochondrial function and cognition in future studies. Methods/Study Population: Peripheral blood mononuclear cells were isolated from virally suppressed WWH (n = 64) and underwent the Seahorse Cell Mito Stress test to assess different realms of mitochondrial function. Cells were then lysed for direct DNA extraction, and quantitative PCR was performed to understand mitochondrial DNA expression (mtDNA) levels as a measure of oxidative stress. A series of simple linear regressions was then conducted to understand the relationships between mitochondrial function and mtDNA content. Future work will expand this analysis to investigate associations between demographic dynamics, such as trauma history, and mitochondrial function, as well as to understand the relationships between mitochondrial function and cognitive outcomes in WWH. Results/Anticipated Results: In our cross-sectional analysis of mitochondrial dynamics in WWH, we found a significant association between maximum mitochondrial respiration ability and mtDNA content, with greater mtDNA expression associated with increased levels of maximum respiration following stimulation. There was no association between basal respiration and levels of oxidative stress. There was also a significant variation in mitochondrial function in our participants, indicating that future analyses to investigate the source of that variation are warranted. The work presented here sheds light on mitochondrial dynamics in WWH and will be the basis for future studies that will investigate how demographic dynamics may be associated with mitochondrial function, as well as how mitochondrial dynamics may predict cognitive outcomes. Discussion/Significance of Impact: There is significant variation in mitochondrial function in WWH. More analysis is needed to understand what may be associated with these variations, including an investigation of both clinical factors as well as cognitive outcomes. This analysis will inform directions for future mechanistic work aimed at mitigating adverse cognitive outcomes in WWH.

Objectives/Goals: This study aimed to identify preferences for nutrient-dense foods with critical nutrients associated with reduced neuroinflammation and perinatal depression risk. Aim two evaluated the acceptability of daily intake and preparation methods of select foods in Black American pregnant and postpartum people living in food apartheid in St. Louis. Methods/Study Population: The study included a mixed-methods sequential explanatory design with data collection at the YWCA of Metro St. Louis during Early Head Start programming with 21 participants living in geographies with low grocery-to-convenience store ratios. We conducted an open-ended online survey to determine participants’ preferences for specific foods with high nutrient density. Food preferences were subjected to nutrient analyses using FoodData Central, and foods with the highest concentrations of critical nutrients associated with reduced neuroinflammation and perinatal depression risk were identified. We presented results during a focus group with participants who completed the survey and evaluated the acceptability of daily intake and preparation methods. Results/Anticipated Results: Preferred food choices with the highest concentrations of critical nutrients associated with reduced neuroinflammation and perinatal depression risk included dark red kidney beans, black beans, pinto beans, black-eyed peas, peanuts, almonds, pistachios, walnuts, pecans, carrots, spinach, red bell pepper, sweet potatoes, and salmon. Participant justifications for low acceptance of daily intake included allergic reactions, “dry” or “slimy” texture, undesirable taste or appearance, absence of cravings, preferring alternatives, complicated preparation methods, and financial tradeoffs. Participant justifications for high acceptance included desirable taste and texture, diverse modification and preparation options, and nutrient composition critical for anemia treatment. Discussion/Significance of Impact: The study identified nutrient-dense food preferences within a sample vulnerable to nutrient-poor alternatives and health consequences. Participant acceptance of nutrient-dense intake was contingent upon recipes fulfilling cravings, acceptance among persons preparing meals, and diverse options to prevent boredom with preparation methods.

Objectives/Goals: We assessed the feasibility of using a large language model (LLM) to create lay language descriptions of study protocols for recruitment, which has the potential to improve accessibility and transparency of clinical studies and enable participants to make informed decisions. Methods/Study Population: All studies from a clinical research recruitment platform were included, which features human-written lay descriptions and titles for study recruitment. Corresponding protocol summaries in the IRB system were extracted and translated into lay language using a LLM (gpt-35-turbo-0613). A subset was used to develop prompt variations through an iterative process. Prompt strategies evaluated include chain-of-thought and few-shot prompting techniques. LLM-generated and human-written descriptions were compared for readability using Flesch–Kincaid and Simple Measure of Gobbledygook (SMOG) reading grade levels and information completeness using Word Movers’ Distance (WMD). Results/Anticipated Results: A total of 55 study descriptions were included – 10 were used to develop prompts and 45 were used for evaluation. The final LLM instructions included multistep prompts. The LLM was first instructed to produce a two- to three-sentence long description without using scientific jargon and included two pairs of examples. The LLM was then asked to shorten the description and finally to provide an engaging title. LLM-generated and human-written summaries were similar in length (median (IQR) 328 (278.5–360.5) vs. 342 (203–532.5) characters, respectively). LLM-generated summaries had lower Flesch-Kincaid grade level (5.15 vs. 8.28, p Discussion/Significance of Impact: An LLM can be used to generate lay language summaries that are readable at a lower grade level while maintaining semantic similarity. This approach can be used to improve the drafting of summaries for recruitment, thereby improving accessibility to potential participants. Future work includes human evaluation and implementation into practice.

Objectives/Goals: Manual skin assessment in chronic graft-versus-host disease (cGVHD) can be time consuming and inconsistent (>20% affected area) even for experts. Building on previous work we explore methods to use unmarked photos to train artificial intelligence (AI) models, aiming to improve performance by expanding and diversifying the training data without additional burden on experts. Methods/Study Population: Common to many medical imaging projects, we have a small number of expert-marked patient photos (N = 36, n = 360), and many unmarked photos (N = 337, n = 25,842). Dark skin (Fitzpatrick type 4+) is underrepresented in both sets; 11% of patients in the marked set and 9% in the unmarked set. In addition, a set of 20 expert-marked photos from 20 patients were withheld from training to assess model performance, with 20% dark skin type. Our gold standard markings were manual contours around affected skin by a trained expert. Three AI training methods were tested. Our established baseline uses only the small number of marked photos (supervised method). The semi-supervised method uses a mix of marked and unmarked photos with human feedback. The self-supervised method uses only unmarked photos without any human feedback. Results/Anticipated Results: We evaluated performance by comparing predicted skin areas with expert markings. The error was given by the absolute difference between the percentage areas marked by the AI model and expert, where lower is better. Across all test patients, the median error was 19% (interquartile range 6 – 34) for the supervised method and 10% (5 – 23) for the semi-supervised method, which incorporated unmarked photos from 83 patients. On dark skin types, the median error was 36% (18 – 62) for supervised and 28% (14 – 52) for semi-supervised, compared to a median error on light skin of 18% (5 – 26) for supervised and 7% (4 – 17) for semi-supervised. Self-supervised, using all 337 unmarked patients, is expected to further improve performance and consistency due to increased data diversity. Full results will be presented at the meeting. Discussion/Significance of Impact: By automating skin assessment for cGVHD, AI could improve accuracy and consistency compared to manual methods. If translated to clinical use, this would ease clinical burden and scale to large patient cohorts. Future work will focus on ensuring equitable performance across all skin types, providing fair and accurate assessments for every patient.

Objectives/Goals: Retention in care is vital for people living with HIV. We used human-centered design (HCD) to engage a community-based research panel over a 5-year period, allowing us to incorporate their insights on research guidance and interpretation of findings to investigate correlates of HIV care outcomes. Methods/Study Population: We recruited a diverse panel of individuals who were living with HIV, HIV clinicians, and/or providing non-clinical HIV services in Marion County, Indiana. We conducted biannual sessions using a variety of HCD tools and activities to engage participants. Each session took about three hours, and panelists were compensated for their participation. Due to the COVID-19 pandemic, sessions were initially held virtually. Sessions were designed for project discussion and to facilitate exploration of concerns and challenges facing receipt of HIV services. Our HCD approach put participants in the center of discussion and empowered them to externalize ideas and collaborate meaningfully with our team. Results/Anticipated Results: Since project inception, 48 individuals have joined the panel. Thirty-five are actively engaged, participating in one or more of six sessions conducted to date. We have learned much from the panel. One example is that a residential move might be a risk or protective factor for retention in care and the amount of time one had lived with HIV is a crucial factor. Panel insights have helped guide and prioritize analyses, aided in identification of data missing from our ecosystem, helped interpret results, provided feedback on future interventions, led to a quality improvement project with the local health department, and led to a presentation at a local health equity conference. Discussion/Significance of Impact: Community engagement is essential to impactful and sustainable research. HCD was a successful approach to engage our panel to inform interventions more relevant to the community. We anticipate these methods will be important for others conducting community-engaged research.

Objectives/Goals: The Wake Forest Clinical and Translational Science Institute (CTSI) has integrated academic goals of T0-T4 translation, scholarship, and education into our Academic Learning Health System (aLHS) framework. Our Translation Research Academy (TRA) provides rigorous training for outstanding and diverse K12 and early-career faculty to develop LHS core competencies. Methods/Study Population: The TRA Forum is the main vehicle for delivering an aLHS-oriented curriculum. Currently, the program includes six K12 scholars and 18 other early-career research faculty with facilitated access to CTSI resources. The TRA Forum is a 2-year seminar series that meets twice a month to discuss topics relevant to the aLHS, leadership, and career development. Inclusion of first- and second-year scholars facilitates peer mentorship, allowing Year 2 scholars to share insights with new scholars. Forum sessions are developed around adult learning theory: Each participant is asked to contribute their experience to discussions, and sessions focus on real-world examples. Results/Anticipated Results: Scholar and faculty commitment is very high. For the first 30 min., scholars present their work in small groups. This extends the range of disciplines exposed (64% of TRA graduates found this very helpful) and promotes translational traits of boundary crosser, team player, and systems thinker. Participants view the TRA as an opportunity to form internal peer networks, promote peer mentoring, and establish new collaborations. The remaining 60 minutes are used for education. Sessions include nominated topics and those providing a solid foundation in core aLHS competencies and characteristics of translational scientists. Educational sessions (97%) were rated as helpful or very helpful. Discussion/Significance of Impact: TRA scholars receive rigorous training in a highly supportive environment to produce aLHS researchers with skills to transcend boundaries, innovate systems, create new knowledge, and rigorously evaluate results.