Objectives/Goals: NAD+ synthesis is enhanced in glioblastoma (GBM) allowing GBM to resist chemotherapy. NQO1 is upregulated in GBM and may be targeted by β-lapachone (β-lap) to induce NAD-depletion and cell death. This project investigates NQO1 as a selective target for GBM and the contributions of glucose and uridine to NAD+ synthesis. Methods/Study Population: Survival Studies and NQO1 expression. RNA-seq and survival data from TCGA of glioma patients (n = 667) was obtained using the UCSC Xena platform. Western blots were utilized to determine expression levels of NQO1 and NAMPT in normal human astrocytes, U87 cells, and patient-derived GBM cell lines. Immunocytochemistry: γ-H2AX staining was used to evaluate β-lap induced DNA damage. NQO1-dependence was evaluated with the NQO1-inhibitor dicoumarol. Cytotoxicity measurements. Cells were exposed to β-lap and other inhibitors, and cell survival was determined by trypan-blue exclusion assay. Co-culturing experiments were performed with fluorescently labeled U87 cells and unlabeled astrocytes. NAD+ quantification. Intracellular NAD+ was acid extracted and quantified by an enzyme-cycling reaction. Results/Anticipated Results: NQO1 overexpression is linked to decreased survival in glioma patients. In glioma patients, high NQO1 expression was associated with a decreased overall survival and high-grade tumors. β-lap induces selective NAD+ depletion and cell death in NQO1-expressing GBM cells. Western blots demonstrate NQO1 expression to be elevated in GBM cell lines compared to normal human astrocytes. β-lap induces NQO1-dependent NAD+ depletion and cell death in GBM compared to astrocytes in mono- and co-culture experiments. Glucose and uridine facilitate NAD+ regeneration in GBM. We demonstrate extracellular glucose and uridine facilitate NAD+ regeneration and cell survival in β-lap exposed GBM cells. Utilizing inhibitors, we determined that glucose and uridine facilitate NAD+ regeneration through the NAD+ salvage pathway. Discussion/Significance of Impact: GBM is the most common primary adult CNS tumor with a median survival of 14 months. Despite significant research in therapeutic strategies, treatment has not improved in 2 decades. There is a significant need to discover new targets that may improve GBM treatment. We demonstrate here that targeting NQO1 with β-lap induces selective GBM toxicity.

Objectives/Goals: The growing burden of hypertension in the USA disproportionately impacts individuals with lower socioeconomic status and those in rural communities. This study aims to investigate specific social determinants of health associated with uncontrolled hypertension among rural, underserved patients residing in Alabama. Methods/Study Population: A retrospective review of electronic health records data from the UAB Selma Family Medicine clinic was conducted. The deidentified data were entered into the Population Health Assessment Engine (PHATE) to render social characteristics and community vital sign (CVS) scores, a social deprivation index, by census tracts. We provide descriptive statistics and compare the stated factors between patients with controlled and uncontrolled hypertension using logistic regression models. We are conducting phone surveys to gather individualized data on social determinants of health to assess the perceived social needs of patients with hypertension. The Protocol for Responding to & Assessing Patients’ Assets, Risks & Experience tool was used to develop the survey, and survey responses will be compared with the PHATE results. Results/Anticipated Results: Among 2129 patients, 375 (17.6%) had uncontrolled hypertension with a mean age of 58.7 ± 14.4 years. Women comprised 66.7% (250) and 65.1% (1142) of the uncontrolled and controlled hypertension populations, respectively. The prevalence of uncontrolled hypertension was 19.1% among Black patients and 8.2% in White patients. Patients with uncontrolled hypertension had a CVS of 82.37 ± 7.31 and 81.81 ± 8.40 patients with controlled hypertension. Patients with uncontrolled hypertension lived in areas where an average of 46 ± 11% were not employed, 36 ± 9% rented, and 28 ± 1% lived below the federal poverty line (FPL). Adjusting for age, race, and sex, a unit increase in the proportion of people below the FPL was associated with 43% higher odds of uncontrolled versus controlled hypertension (OR  =  1.43, 95% CI 0.44, 4.64). Discussion/Significance of Impact: Rural, underserved patients with hypertension experience multiple adverse social determinants, such as nonemployment rates, low income, and rental housing. Survey results will allow us to compare assessed and individualized social determinants that may be important to address when caring for this population.

Objectives/Goals: To design and implement programming that better prepares graduate students for diverse roles in a variety of workforce environments, our study models the training landscape and programming needs of graduate students in behavioral, clinical, and biomedical graduate programs at a large Midwestern school of medicine and public health. Methods/Study Population: We conducted six focus groups (two graduate program manager focus groups and four graduate student focus groups), to assess the programming, career development, and training needs of graduate students. Using a grounded theory approach, we first engaged in open coding of a sample of transcripts. After developing a codebook, we continued with an iterative coding process interspersed with coder meetings to discuss emerging and changing codes. Using the framework of landscape analysis allowed our coding and modeling to go beyond graduate student needs and study the varying relationships and contexts that impact graduate students throughout their training, such as relationship to supervisor or institutional policies. Results/Anticipated Results: Preliminary results indicate that students wrestle with their status as both students and workers. Specifically, conflict arises between graduate and supervisor expectations around time spent in class, lab, and other career development activities based on these divergent roles. Students and program managers also note the disparities that arise from the university’s lack of standard, formalized policy on labor issues, such as paid leave. Data also suggest that students on training grants note the difference in access to career development resources compared to colleagues. In many cases, students themselves coordinate ad hoc programming to better suit their career and professional development needs, although this work is not a required aspect of their training. Discussion/Significance of Impact: We characterize current graduate training landscapes, which continue to shift as graduate student bodies diversify, unionize, and express interest in increasingly varied biomedical careers. Data from multiple perspectives facilitate creating, implementing, and evaluating supportive training programs that meet identified student needs.

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: This project will enrich our understanding of basal breast carcinogenesis, highlighting the distinct biological differences in breast cancer risk between non-Hispanic Black (NHB) or non-Hispanic White (NHW) women, but also has the potential to provide real-world solutions thereby contributing to reducing health disparities in breast cancer outcomes Methods/Study Population: In the biological investigation, I am utilizing a unique cohort of normal breast tissues from Mayo Clinic patients who underwent breast reduction surgery that have self-identified as NHB or NHW premenopausal women. From these tissues, we are able to subculture human mammary epithelial cells, which we will use for our consequent experiments. For the qualitative study, I aim to recruit up to 40–50 eligible NHB women identified at increased risk for breast cancer and conduct semi-structured qualitative interviews informed by the National Institute on Minority Health and Health Disparities Research Framework. Results/Anticipated Results: Gene expression profiling on our cohort of 15 NHB and 40 NHW premenopausal women identified a gene expression signature in NHB women indicative of elevated Hedgehog signaling, a key factor in triple-negative breast cancer (TNBC) development and progression. I hypothesize that persistent activation of Hedgehog signaling within NHB women instigates the proliferation and transformation of basal stem cells within the breast, thus fueling TNBC development. Understanding the perceptions and experiences of NHB women that are identified as being at an increased risk for BC will aid in the identification of barriers and facilitators to their risk reduction care and in turn provide implementation strategies that could contribute to alleviating the racial disparity seen in TNBC morbidity, mortality, and preventative care. Discussion/Significance of Impact: The significance of this project lies in its dual approach, uncovering basal breast cancer mechanisms in a high-risk group using normal breast tissue and evaluating the perspectives from NHB women to enhance the practical relevance of our findings.

Objectives/Goals: Micro-credentials (MCs) or digital badges are short programs designed to allow learners to gain knowledge and skills at their own pace to tailor their professional development. This study aims to examine the characteristics of learners completing MCs and explore their motivation for pursuing MCs through the University at Buffalo Clinical and Translational Science Institute (CTSI) program. Methods/Study Population: Currently, the CTSI offers five MCs in Effective Teaching, Good Clinical Practice, Responsible Research, Scientific Communication, and Translational Teamwork. Individuals who completed an MC (2019–2024) were identified by the UB Office of Micro-credentials. An invitation email and two reminders were sent to all individuals who received MCs asking them to complete a short online survey in July-August 2024. The survey included three questions about the type(s) of MCs completed, learners’ motivation for pursuing MCs, and perceived or actual benefits of completing an MC. The questions included multiple choice, select all that apply, and open-ended format, respectively. The survey was sent using Google Forms; data were analyzed using descriptive statistics. We received 25 completed surveys (29% response rate). Results/Anticipated Results: Since 2019, 85 individuals completed MCs at Buffalo CTSI including 21 faculty, 18 trainees, 26 students, and 20 staff. The most popular MC, Responsible Research (65% of completers), is based on the Responsible Conduct of Research series, which consists of modules on authorship ethics, ethical use of animals (IACUC) and human subjects (IRB), conflicts of interest/commitment, and responsible data acquisition and management. We found that 33% of responders were motivated to obtain the MC to advance their career and for professional development, 30% for lifelong learning, and 23% intended to use the badges to showcase their proficiency to potential employers. The greatest benefits reported were to obtain knowledge/skills for their professional career (46%) and improve their ability to receive research funding (14%). Discussion/Significance of Impact: Micro-credentials are valuable learning tools to stay current on changing research requirements, with ability to engage asynchronously. Badges also provide unique professional development opportunities for students and research workforce (staff and community stakeholders) who have limited resources (time and money).

Objectives/Goals: The never in mitosis kinase (NEK) family regulates vital processes, namely cell cycle progression, but their potential as therapeutic targets in TNBC has not been fully explored. Our studies aim to develop a toolkit to investigate the functional roles of NEKs in pathologies including carcinogenesis. Methods/Study Population: To assess differential NEK expression in normal and tumor tissues and correlation of gene expression with patient survival, we used Gene Expression Profiling Interactive Analysis (GEPIA) and Kaplan–Meier Plotter (KMPlot) pan-cancer analysis, respectively. Basal NEK protein levels were determined by immunoblot across a panel of cell lines, including breast cancer, osteosarcoma, hepatocellular carcinoma, and non-cancerous cells, to identify appropriate systems for evaluation of NEK function. Doxycycline-inducible cell lines were generated by transduction with lentiviral stocks of NEK shRNA and overexpression constructs and antibiotic selection. Expression was analyzed by qPCR and immunoblot. Results/Anticipated Results: Expression of NEK2, 4, 5, 6, 8, and 11 was higher in breast tumors compared to normal tissue by GEPIA analysis. Further examination using KMPlot showed a correlation between elevated NEK6 expression and decreased overall survival in patients with aggressive cancers. As an initial proof-of-concept study, we analyzed NEK6 protein expression in breast cancer cells. Levels of NEK6 were elevated in TNBC cells (MDA-MB-231) compared to hormone receptor positive (HR+) breast cancer cells (MCF7). Using complementary approaches to investigate the functional role of NEK6 in breast cancer, we depleted NEK6 expression using shRNAs in TNBC cells and expressed NEK6 in HR+ cells Discussion/Significance of Impact: Because kinase dysregulation promotes oncogenesis and metastasis, targeting kinases is a key strategy in therapeutic development. A NEK-specific molecular toolkit allows researchers to elucidate NEK functions and contributions to carcinogenesis, promoting advancement of novel therapies.

Objectives/Goals: The creatine (Cr) system is impaired in Alzheimer’s disease (AD). Data show that creatine monohydrate (CrM) supplementation may improve AD symptoms in AD mouse models, but no human studies have been reported. Thus, we investigated whether an eight-week CrM supplementation was feasible and associated with increased brain creatine in patients with AD. Methods/Study Population: Twenty participants with probable AD were allocated to an open-label, eight-week intervention of 20 g/day CrM. Fasting blood draws were taken at baseline, 4-, and 8-week visits to measure serum creatine (Quest Diagnostics). 1H magnetic resonance spectroscopy was performed at baseline and 8-week visits to measure brain Cr as a ratio to unsuppressed water. Self-reported compliance (with assistance from study partners) was assessed with daily CrM trackers. The mean compliance percentage across all participants was used to describe overall compliance with the intervention. We used paired t-tests to analyze the mean changes in serum Cr levels from baseline to 4- and 8-week visits and the mean change in brain Cr from baseline to 8-week visits. Statistical significance was set at p<0.05. Results/Anticipated Results: Participants were 65% male with a mean age of 73.1±6.3 years. All participants completed the study, with 19 out of 20 achieving the dose compliance target of ≥80%. The mean self-reported dose intake was 90%. Serum Cr levels were significantly increased at 4- and 8-week visits compared to baseline (0.6±0.4 mg/dL vs. 14.0±9.9 mg/dL and 15.0±13.6 mg/dL, respectively; p<0.001). Brain Cr levels also significantly increased (330.5±36.80 i.u. vs. 366.9±57.52 i.u., p<0.001). Discussion/Significance of Impact: We are the first to demonstrate that 20 g/day of CrM for eight weeks is feasible and associated with increased brain Cr in patients with AD. Our findings support further investigation of brain target engagement of CrM and its efficacy in AD. With AD cases expected to rise, CrM could serve as an effective, affordable therapeutic to slow AD progression.

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: A key strategy in generating a protective HIV vaccine is the elicitation of broadly neutralizing antibodies (bnAbs), capable of neutralizing a large diversity of HIV-1 isolates. The goal of this study is to identify molecular signatures of HIV bnAb development early in life, to guide the development of a successful pediatric HIV vaccine strategy. Methods/Study Population: We previously defined HIV neutralization breadth in 40 ART-naive children living with HIV. Single-cell RNAseq was performed utilizing peripheral blood mononuclear cells (PBMCs) from the top 5 children with highest neutralization breadth scores and compared their transcriptome to that of PBMCs from 5 children that did not develop neutralization breadth within the first three years of life. Additionally, we incorporated analysis of PBMCs from 5 healthy uninfected children, matched to our experimental groups by race, ethnicity, and gender. Results/Anticipated Results: We expect that a distinct host transcriptional profile is associated with the development of HIV-specific antibody neutralization breadth in early life. Discussion/Significance of Impact: Identifying immune cell transcriptional profiles associated with neutralization breadth will lead to more targeted vaccine approaches for eliciting the appropriate B cell responses and provide an invaluable screening tool allowing early identification of vaccine candidates with the potential to induce bnAbs.

Objectives/Goals: Our research group is focused on care of hospitalized persons with opioid use disorder (OUD) in the era of high-potency synthetic opioids (HPSO). In this work, we describe trends in patient-directed discharge (PDD) and inpatient treatment with medications for opioid use disorder (MOUD). We hypothesized that PDD is associated with MOUD dose and timing. Methods/Study Population: Patient data generated in the routine care of patients was automatically abstracted using a SQL query on Epic Clarity tables in the electronic health record (EHR). We included adult patients admitted to Johns Hopkins Hospital between July 1, 2019 and June 30, 2022, with an ICD-10-CM code for a list of opioid-related disorders (F11.X) consistent with OUD. Demographics, prior medication list, clinical care including hospital service, consultation services, COWS scores, length of time in emergency department, time of triage, time until receipt of methadone or buprenorphine, dosage and timing of MOUD, opioid medications other than methadone or buprenorphine, adjuvant medications; prior methadone or buprenorphine treatment and disposition. Query results were validated by manual abstraction of EHR. Results/Anticipated Results: The SQL identification of the cohort of patients with OUD was found to be accurate. Time of triage, discrete orders completed during hospitalization were well represented in the query. The query was able to identify individual opioid medications but unable to summarize total dose in Morphine Milligram Equivalents. The query did not extract accurate information from patient-controlled analgesia pumps due to the continuous nature of the medication rather than discrete doses reflected in the medication administration record. Finally, the query characterized prior treatment with methadone or buprenorphine as a binary variable – dosage and timing of that prior treatment could not be accurately represented. Finally, stimulant use is not reliably collected in the EHR and was unavailable. Discussion/Significance of Impact: Given the rise of HPSO, patients may not tolerate delay of MOUD. Improving the granularity of data collected will offer more insight into the inpatient treatment for OUD. Real-world data have promise but requires extensive technical expertise. Future work is needed to improve capture of derived variables such as total dosage of opioids in MME.

Objectives/Goals: Our objective is to evaluate how access to oral health care and tobacco use in patients with serious mental illness (SMI) impact their oral health outcomes identifying barriers and service utilization disparities affecting the prevalence and severity of oral diseases. Methods/Study Population: A secondary retrospective data analysis study will use records from the electronic database (All of Us). The study population includes individuals over 21 years old, diagnosed with SMI. Data will be collected focusing on their oral health status, access to oral health care (frequency of visits, availability of services, etc.), tobacco use, and social determinants of health. The study will evaluate the impact of access to oral health care on the prevalence of oral health outcomes (e.g., periodontal diseases and oral cancer), analyzing socioeconomic factors, mental health status, and tobacco use as independent variables. A quantitative, descriptive-correlational analysis will be performed to explore the study variables. Results/Anticipated Results: We anticipate that limited access to oral health care, influenced by socioeconomic factors, the presence of SMI, and tobacco use, will correlate with worse oral health outcomes in Hispanics. Disparities in access, such as financial limitations, transportation issues, and stigma, are expected to significantly hinder service utilization and increase the prevalence of oral diseases in patients with SMI who smoke. Discussion/Significance of Impact: This research highlights the need to address disparities in oral health access for Hispanic marginalized populations, particularly those with severe mental illness and tobacco use. It will contribute to reducing health inequalities and improving oral health outcomes through culturally adapted interventions that address these barriers.

Objectives/Goals: Global surgical education is largely driven by high-income countries (HICs), with curricula not tailored to the needs of low- and middle-income countries (LMICs). This study assessed country-specific needs for global surgical curricula and used generative AI to develop tailored curricula. Methods/Study Population: A curriculum framework was developed using expert opinion. Using a focused needs assessment survey, we evaluated international medical students’ and trainees’ needs for structured global surgery curricula, covering research, education, data and develop tailored curriculum templates for each country, ensuring alignment with the distinct needs of respective LMIC and HIC respondents. The AI-generated curricula were then compared across countries to identify variations in content and focus areas. Results/Anticipated Results: A total of 145 respondents from 18 countries and 6 continents participated, with 94 from LMICs and 51 from HICs. Four countries [Uganda (n = 31), Nigeria (n = 34), the USA (n = 23), and the UK (n = 23)] had more than 10 respondents, with the creation of a country specific global surgery curriculum. Curricula developed by HIC trainees focused on access to resources and infrastructure, future directions of global surgical research, and the role of medical students and early career development with a decreased focus on the history of global surgery. LMIC country-based curriculum focused on introducing the concepts of global surgery, quantifying the burden and epidemiology of surgical disease and had a greater emphasis on case studies and use cases, with decreased focus on resources and collaboration. Discussion/Significance of Impact: The research introduces a “precision education” approach that could help close the surgical education access gap globally. Further pilot and qualitative studies are necessary to validate the feasibility of AI-generated needs-based curricula.

Objectives/Goals: The clinical research professional (CRP) workforce suffers from high turnover. Stay interviews have led to increased satisfaction and reduced turnover in other industries. We describe a multi-institutional project to develop, disseminate, and evaluate a CRP-tailored Stay Interview tool reimagined as the Individual Retention Conversation (IRC) toolkit. Methods/Study Population: In August 2022, following on the heels of a series of un-meeting conversations focused on CRP workforce development, the CRP taskforce initiated a working group to tackle issues related to CRP workforce retention. As a first initiative, this multi-institutional working group set out to develop, disseminate, and evaluate a Stay Interview tool tailored for a CRP audience and reimagined as the IRC toolkit. A 2-phase pilot study was initiated across six academic medical centers (AMCs: ASU, Duke, MUSC, UAB, UPenn, VCU) to: 1) optimize the toolkit for the CRP audience and 2) evaluate the impact of the toolkit using a standardized CRP satisfaction survey. Quantitative and qualitative data were collected via surveys using the REDCap platform. Results/Anticipated Results: The optimization phase of the pilot included 69 participants (16 managers and 53 of their CRP team members) from 6 AMCs. Respondents identified most and least useful questions for stimulating meaningful conversations regarding job satisfaction and retention issues with additional feedback on the IRC experience and tools. CRPs and managers represented a variety of roles, with 77% patient facing. The majority were satisfied with the IRC experience (82%) and found the experience personally beneficial (76%). Managers were satisfied with the manager’s guide (90%). Quantitative and qualitative feedback was used to optimize the toolkit prior to launch of phase 2 in September 2024, which includes a longitudinal survey-based assessment of CRP job satisfaction and IRC-consequent work environment changes. Discussion/Significance of Impact: CRP retention is impacted by complex factors, many related to job satisfaction, supervisor /employee relationships, and beneficial work environments. Initial evaluation of the IRC suggests that this intervention fosters positive supervisor/employee relationships and beneficial work environment changes, which may lead to improved retention.

Objectives/Goals: Predictive performance alone may not determine a model’s clinical utility. Neurobiological changes in obesity alter brain structures, but traditional voxel-based morphometry is limited to group-level analysis. We propose a probabilistic model with uncertainty heatmaps to improve interpretability and personalized prediction. Methods/Study Population: The data for this study are sourced from the Human Connectome Project (HCP), with approval from the Washington University in St. Louis Institutional Review Board. We preprocessed raw T1-weighted structural MRI scans from 525 patients using an automated pipeline. The dataset is divided into training (357 cases), calibration (63 cases), and testing (105 cases). Our probabilistic model is a convolutional neural network (CNN) with dropout regularization. It generates a prediction set containing high-probability correct predictions using conformal prediction techniques, which add an uncertainty layer to the CNN. Additionally, gradient-based localization mapping is employed to identify brain regions associated with low uncertainty cases. Results/Anticipated Results: The performance of the computational conformal model is evaluated using training and testing data with varying dropout rates from 0.1 to 0.5. The best results are achieved with a dropout rate of 0.5, yielding a fivefold cross-validated average precision of 72.19% and an F1-score of 70.66%. Additionally, the model provides probabilistic uncertainty quantification along with gradient-based localization maps that identify key brain regions, including the temporal lobe, putamen, caudate, and occipital lobe, relevant to obesity prediction. Comparisons with standard segmented brain atlases and existing literature highlight that our model’s uncertainty quantification mapping offers complementary evidence linking obesity to structural brain regions. Discussion/Significance of Impact: This research offers two significant advancements. First, it introduces a probabilistic model for predicting obesity from structural magnetic resonance imaging data, focusing on uncertainty quantification for reliable results. Second, it improves interpretability using localization maps to identify key brain regions linked to obesity.

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: 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: 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: 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: 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.