Objectives/Goals: Despite the documented link between ultraviolet (UV) exposure and skin cancer, the potential protective role of pigmentation in preventing skin carcinogenesis is not fully understood. This project will test the hypothesis that individuals with light skin tones are more susceptible to skin cancer due to a higher accumulation of somatic mutations from UV exposure. Methods/Study Population: Skin biopsies were collected from various anatomical sites of seven donors aged 70 to 80 years with dark skin tones and used to generate clones of normal skin cells. The clones were then subjected to whole-exome and RNA sequencing. We developed a workflow that allows us to accurately detect somatic mutations in clonal expansions of individual cells with high specificity and sensitivity. For comparison, we also analyzed mutations in skin cells obtained from individuals with light skin tones, who served as our control group. In the future, we plan to employ admixture analysis to clarify the impact of race on somatic mutations by examining ancestral genetic contributions. Results/Anticipated Results: Our preliminary analysis of the existing data supports our hypothesis. The average mutation burden in individuals with light skin tone was found to be 3.778 Mut/Mb, whereas in those with dark skin tones, was only 0.818 Mut/Mb. These findings align with the prevailing hypothesis that melanin acts as a protective factor against skin cancer, as it appears to reduce the mutagenic effects of UV radiation. This suggests that individuals with darker skin may have a lower cumulative mutational load, potentially contributing to their reduced risk of developing skin cancers such as melanoma. Similarly, our results showed variations in mutational burdens across different anatomical sites, which seemed to be influenced by differing levels of UV exposure, with the highest burdens observed in areas with the greatest exposure. Discussion/Significance of Impact: Understanding how skin cancer operates in darker-skinned individuals is imperative for tailoring effective screening and targeted therapies to meet their specific needs. In the long run, elucidating the mechanisms underlying skin cancer development in this demographic will help refine screening protocols and prevention recommendations.

Objectives/Goals: To reduce cervical cancer in the USA, it is essential to identify the reasons underlying the low HPV vaccination rates. This study aims to identify knowledge and beliefs about HPV vaccination among women in rural areas. The knowledge gained from this study will directly lead to developing an education tool tailored specifically for women in rural areas. Methods/Study Population: We conducted a cross-sectional study from November 2022 to July 2023. We recruited women (n  =  141) who visited a mobile health unit in rural North Louisiana. The inclusion criteria were women aged 25–64 years who spoke English, had not undergone a total hysterectomy, and had no history of cervical cancer. Data collected included sociodemographic characteristics, sexual history, awareness and knowledge of HPV infection, cervical cancer, genital warts, and HPV vaccination, the perceived risk of acquiring genital HPV infection and developing cervical cancer, and the willingness to receive an HPV vaccine. Descriptive statistics were used to evaluate participant responses. Written informed consent was obtained before completing the self-administered questionnaire. Results/Anticipated Results: Our findings showed significant gaps in HPV vaccine knowledge and uptake. Approximately 40% of the participants were unaware of the HPV vaccine, 96.5% had never received it, and 91.4% had never been offered it. However, 42% indicated a willingness to consider vaccination if it were offered. Factors influencing their decision to vaccinate against HPV included a family history of cervical cancer (44.7%), having multiple sexual partners (48%), and engaging in unprotected sex (61.7%). Furthermore, there was significant uncertainty surrounding the vaccine, with 65.2% of participants unsure about its safety, 66% unaware of potential health risks, and 47.5% uncertain about its effectiveness in preventing HPV. Discussion/Significance of Impact: Our research emphasizes the need for tailored behavioral interventions to address knowledge gaps about the HPV vaccine. Low HPV vaccination rates in rural areas contribute to health disparities in cervical cancer. Implementing educational interventions in healthcare settings can enhance vaccination rates and mitigate the risk of cervical cancer.

Objectives/Goals: Obesity has been classified as a global epidemic and origin of numerous health issues. The central hypothesis of this study is that psychological measures, DNA methylation, and gene transcription will predict obesity-related outcomes after lifestyle interventions, and such interventions may alter DNA methylation profiles. Methods/Study Population: This study consisted of a randomized-controlled trial examining the effects of lifestyle +/- stress reduction interventions in 285 highly stressed parents with obesity, followed for 2 years. Full participants received nutrition and activity counseling, and were randomized to either a stress reduction intervention or a contact control. Those who otherwise qualified for the study but unable to fully participate were included in a no intervention control group. The intervention consisted of 12 weeks of nutrition and activity counseling +/- 2-hour weekly stress reduction interventions using MBSR and CBT-based strategies. DNA methylation was assessed using Illumina EPIC arrays. Results/Anticipated Results: Using linear mixed models (LMMs), this study will first examine the hypothesis that baseline psychological measures and pre-existing methylation sites associated with obesity and glycemic control (e.g., ABCG1, ATP10A, TXNIP, SREBF1, RNF39, and SOCS3) predict changes in BMI, HOMA-IR, and HgbA1C post-intervention and at 1 and 2 year follow-ups. Using sites that demonstrate statistical significance, we will develop a polymethylation risk score predictor of change in BMI. Next, we will examine the hypothesis that interventions which reduce obesity may also lead to improvements in epigenetic aging using LMMs to determine if changes in BMI or HOMA-IR predict changes in epigenetic age acceleration over the course of the study. Discussion/Significance of Impact: This work examines whether psychological factors and/or epigenetic markers may be used in patient stratification at initiation of treatment, enabling improved treatment selection, fewer years of obesity and decreased risk of comorbidities. This proposal also asks whether lifestyle interventions impact the aging process itself.

Objectives/Goals: In mice, it has been shown that loss of Cib2 (calcium and integrin-binding protein 2) results in progressive retinal disease that recapitulates many characteristics of age-related macular degeneration (AMD). This study aims to characterize transcriptional changes in the retinal pigment epithelium (RPE) that underlie this disease process. Methods/Study Population: RPE tissue samples, pooled from 2–3 mice for each biological group, were collected from Cib2-KO and wildtype (WT) mice at two (young) and eight (aged) months of age. Bulk mRNA sequencing was performed using the Illumina HiSeq 4000. Reads were aligned to the UCSC mouse reference genome and quantified using HTSeq. Significant differentially expressed genes (DEGs) between mouse genotype and age groups were assessed using DESeq. CLICK unsupervised clustering followed by gene ontology analysis was performed to identify cellular processes and molecular pathways affected by loss of Cib2 as well as age. Results/Anticipated Results: CLICK analysis revealed several functional pathways that are differentially expressed between sample groups. For example, in both young and aged mice, pathways upregulated in Cib2-KO samples included calcium signaling, RhoA signaling, and integrin signaling. Uniquely downregulated DEGs in young Cib2-KO animals were related to complement and coagulation cascades, LXR/RXR activation (related to lipid synthesis and transport), and phagosomes. Aged Cib2-KO mice displayed the most significant downregulation of genes in the phototransduction pathway, indicating temporal changes in functional pathways that correlate with disease progression. Next steps in analysis include investigating patterns in RPE- and AMD-signature gene sets that may identify molecular pathways more specific to human disease. Discussion/Significance of Impact: Many current studies investigate the role of complement activation, vesicle trafficking, and ion transport as top contributors to AMD development. We identified DEGs paralleling many of these molecular pathways in Cib2-KO mice, highlighting their potential as a model to study age-related RPE pathologies and evaluate therapeutic interventions.

Objectives/Goals: Monensin is FDA approved for use in veterinary medicine. Recent studies pointed to its potent anticancer activity. Since de novo drug discovery process typically takes 10 to 15 years and requires an investment of approximately $1.3 to $3 billion, drug repositioning can bypass several steps in this process and increase the potential for success. Methods/Study Population: Cell viability assays were conducted on human MDA-MB-231, MDA-MB-468, and MCF10A breast cancer cell lines and mouse EO771 and 4T1 breast cancer cell lines. MDA-MB-231 cell line was used in all the studies unless specified otherwise. Time course levels of Bcl-2, Bak, p62, and LC3II were assessed via Western blotting with GAPDH as a loading control. Proteomics analysis was conducted by the IDEA National Resource for Quantitative Proteomics. Time course levels of major histocompatibility complex (MHC) I and II and calreticulin were evaluated using flow cytometry. At least three biological replicates have been conducted for each experiment. Results/Anticipated Results: Monensin and several of its novel analogs were potent toward human and mouse breast cancer cell lines. Furthermore, they induced apoptotic cell death as evidenced by Annexin V/PI assay, downregulation of Bcl-2, and upregulation of Bak in MDA-MB-231 cells. Proteomics analysis revealed that several molecular pathways related to MHC class I and II antigen presentation were significantly altered following treatment with these compounds. Additionally, monensin and its analogs significantly increased the expression of MHC class I and II. Our studies also showed that monensin and its analogs increase the surface calreticulin levels. Treatment of MDA-MB-231 cells with these compounds also resulted in an increase in p62 and LC3II expression, suggesting a disruption of the autophagic process. Discussion/Significance of Impact: These results suggest that monensin and its analogs not only exhibit anti-breast cancer cell activity but also modulate immune-related pathways. By disrupting autophagy and enhancing calreticulin levels, these compounds may potentiate antitumor immune responses, providing a promising avenue for drug repositioning in cancer therapy.

Objectives/Goals: Lack of comparative data limits research operations quality improvement (QI). The Northwest Participant and Clinical Interactions (NW PCI) Network, a group of 17 unaffiliated university and health system-based research centers, built an operations dashboard to track efficiency and enable multisite QI projects. Methods/Study Population: A Data Governance Working Group was assembled to establish shared data governance, draft nondisclosure (NDA), and data transfer and use agreements (DTUA) suitable across organizations and standardize research operations metric definitions. Sites in the NW PCI Network were recruited to participate in a pilot program to assess data sharing and governance infrastructure, data collection, upload procedures, and data visualization tools. The NW PCI Coordinating Center developed an analytical data dashboard of research operations metrics and conducted semi-structured interviews with participating sites to understand barriers and facilitators of program success. Results/Anticipated Results: Four sites (2 health systems, 2 universities) were recruited for the pilot and reviewed and executed NDAs and DTUAs. Three of the sites have submitted data for a total of 1,405 studies. Of the 24 requested data operations metrics (e.g., study startup, recruitment, implementation, and basic study information), 71% of the metrics were submitted by all three sites (n = 17), 25% were submitted by at least one site (n = 6), and 4% were not submitted by any site (n = 1). Interviews with sites after data submission found areas for improvements (clarification of data definitions, efficiency of data upload process) and positive effects for sites (e.g., process improved insight into own data operations). Discussion/Significance of Impact: Unaffiliated research centers created data governance procedures to enable sharing of operations data. Pilot sites successfully loaded most but not all operations data to the dashboard. Interviews identified process limitations and opportunities for improvement to inform expansion to all NW PCI sites.

Objectives/Goals: Understand the impact of sarcopenia on the main respiratory muscle, the diaphragm (DIAm). We hypothesize that in the DIAm of older (i.e., 24 months) compared to younger (i.e., 6 months) rats, maximum specific force (P0) is reduced, maximum shortening velocity (Vmax) is slower, maximum power output is reduced, and endurance is improved. Methods/Study Population: Mid-costal DIAm strips were excised from 6-month (n = 8; 4 female and 4 male) and 24-month (n = 8; 4 female and 4 male) rats. The DIAm was stimulated using platinum plate electrodes, and mechanical and endurance properties were measured (at 26oC). Results/Anticipated Results: In the DIAm, maximum tetanic formce (P0) decreased by ~35%, maximum velocity of shortening (Vmax) slowed by ~20%, and peak power output was reduced by ~35% in 24-month compared to 6-month rats. During repetitive isovelocity (30% Vmax; approximating peak power output) contractions, endurance (the period during which power output was sustained) of the DIAm was unaffected by aging. Corresponding with previous findings, Discussion/Significance of Impact: The changes in DIAm mechanical performance corresponded to an age-related atrophy of type IIx/IIb muscle fibers. We conclude that force generation and endurance of the DIAm required for breathing motor function is preserved in old age, while DIAm sarcopenia does impair more forceful expulsive airway clearance and voiding behaviors.

Objectives/Goals: Skilled clinical research professionals are essential to efficient and effective research teams, but many undergraduate students are not aware of staff-level careers in the field. To address this, the Michigan Institute for Clinical and Health Research launched the Clinical and Health Research Professional (CHRP) Pathways Program. Methods/Study Population: The 10-week hybrid CHRP Pathways Program was piloted in Summer 2024. Undergraduate students interested in research careers were recruited from local universities and community colleges. The program consisted of 8 hours/week of didactic curriculum guided by ECRPTQ competencies and 32 hours/week of mentored work experience on research teams. Classroom activities were completed with participants from a partner program within the university. Surveys were administered at 3 weeks and 10 weeks along with an end-of-program focus group to assess acceptability and self-reported learning outcomes. Results/Anticipated Results: The pilot CHRP Pathways Program enrolled 3 students and was well-received by participating students and mentor research teams. Students agreed that they would recommend the program to their classmates. Students indicated that the program helped them understand the role of a study coordinator, provided insight into research career paths, and helped them form professional relationships. Self-reported confidence levels in a range of research competencies increased. Two students who completed the 10-week program chose to continue working part-time with their research teams while continuing their undergraduate studies. Discussion/Significance of Impact: Experiences like the CHRP Pathways Program provide valuable exposure to staff-level research career opportunities for students engaged in health science studies. They can address an existing workforce gap by equipping college graduates with relevant work experience and basic research competencies.

Objectives/Goals: Obtaining reliable clinical research professional (CRP) employment data within and across Clinical and Translational Science Awards (CTSA) institutions is an ongoing challenge. We describe an intra-institutional approach implemented to generate routine and accurate CRP data reports to monitor and evaluate CRP career progression and assist in formation of an institutional CRP network. Methods/Study Population: A research job family with 47 job series including human, animal, and laboratory research positions was implemented at Virginia Commonwealth University (VCU). However, CRP job satisfaction surveys and evaluations could not be confidently interpreted due to the confounding animal and laboratory research positions. Led by VCU Clinical and Translational Science Awards Workforce Development a cross-functional team was formed to isolate specific CRP positions. The team included CRP front-line staff and managers partnering with VCU Human Resource Information Systems. Identified were 39 unique CRP positions across 13 distinct job series. This identification provides CRP new hire and job specific data for evaluation and tracking as well as the ability for CRP directed communications. Results/Anticipated Results: Initial and monthly HR data reports were used to develop an institutional CRP list-serv for 325–350 allowing for targeted CRP communications within a decentralized environment. Bimonthly HR data reports identify university new hires and internal transfers into any of the 39 unique jobs within 0 – 12 days of hire. Twelve unique data points are provided (name, email, current position hire date, job code, job title, working title, department, division, supervisor’s name, job title, email, and job code) allowing for tracking and analysis of retention rates, career progression, and lateral movement among other outcomes. Collaboration led by VCU Clinical and Translational Science Awards Workforce Development team provides the representative CRP staff, managers, and institutional leadership with a renewed confidence interpreting CRP employment data. Discussion/Significance of Impact: The team science approach to identify and develop routine and real-time reporting of CRP job specific data provides a rich source of information. The information is used to evaluate CRP job satisfaction and factors contributing to CRP retention, engage in future mixed-methods research, and support the formation of an institutional CRP network.

Objectives/Goals: The goals of this research are to 1) determine the prevalence of perinatal doula services use in Virginia, with a focus on individuals with substance use disorders (SUD), 2) evaluate awareness of doulas among pregnant and postpartum people with SUD, and 3) assess provider knowledge and interaction with doulas for the care of this population. Methods/Study Population: Both quantitative and qualitative methods will be used to evaluate patient and healthcare provider knowledge regarding doula services and the patient–doula–healthcare provider relationship. Surveys and semi-structured interviews will be administered to doulas, pregnant and postpartum women, and healthcare providers in this mixed-methods approach. Information from the Centers for Medicare and Medicaid Services National Provider Identifier (NPI) Registry, and doula training programs will be utilized to recruit doulas for participation. Paper and online recruitment materials will be posted to engage pregnant and postpartum individuals. Healthcare provider recruitment will occur via the NPI Registry along with contacting physicians’ practices. SAS 9.4 and NVivo will be utilized for analysis. Results/Anticipated Results: This proposed research will be an initial assessment of the current state of doula services utilization, mothers’ knowledge of doulas and their purpose, and healthcare providers’ awareness of and partnership with doulas to provide optimal birthing and postpartum experiences to the pregnant and parenting population with and without SUD. Results from this study will be disseminated to community doulas, pregnant people and mothers with substance use disorders, and relevant healthcare providers to decrease barriers to doula care and advocate for consistent, systematic documentation of doula services in the medical record and in public health surveillance systems. Discussion/Significance of Impact: This study will be the first study to assess doula services utilization in Virginia, with a specific focus on pregnant and postpartum women with substance use disorders. This work will support advocacy for improved data capture and utilization regarding doula services in order to reduce barriers to care and improve perinatal outcomes.

Objectives/Goals: Methicillin-resistant Staphylococcus aureus (MRSA) is a human bacterial pathogen and is classified as a serious threat. MRSA has become resistant to most B-lactam antibiotics (penicillins and cephalosporins). The goal of this study is to identify an antibiotic adjuvant capable of resurrecting B-lactams for the treatment of MRSA infections. Methods/Study Population: A fluorescence-reporter assay was used to screen a compound library. Minimum-inhibitory concentrations were assessed for the compounds against various MSSA and MRSA strains. A common resistance mechanism to B-lactams by MRSA is by the function of the bla operon. One gene in this operon encodes for a B-lactam sensor/signal transducer protein BlaR, the primary target of this study. Inhibition of BlaR by compound 1 (best potentiator of oxacillin) was studied by nano-differential scanning fluorimetry (nanoDSF), surface plasmon resonance (SPR), scanning electron microscopy (SEM), and time-kill assays. Results/Anticipated Results: We identified 80 compound hits from a 1,974-compound NCI library. Twenty-four compounds showed potentiating ability (2- to 4,096-fold decrease in MIC for oxacillin). Seven compounds exhibited melting temperature shifts by nanoDSF of BlaR, indicating binding. SPR determined compound 1 has a binding affinity of 31 micromolar to BlaR-SD. SEM images showed disruption in the S. aureus cell wall on exposure to compound 1 and oxacillin. S. aureus N315 showed 3-log reduction in bacterial count treated with a mixture of compound 1 and oxacillin. Discussion/Significance of Impact: Compound 1 targets BlaR-SD, which restores S. aureus susceptibility to treatment by oxacillin. There are currently few antibiotics available in the clinic capable of treating MRSA infections. The combination hold promise of a treatment option for MRSA.

Objectives/Goals: Electronic health record (EHR)-based recruitment can facilitate participation in clinical trials, but is not a panacea to trial accrual challenges. We conducted a root cause analysis to identify EHR-based accrual barriers and facilitators in a pragmatic randomized trial of metformin for those with prostate cancer and glucose intolerance. Methods/Study Population: We quantitatively analyzed enrollment drop-offs among eligible patients who either did not complete a consent (with analysis of EHR-embedded consent process) or who completed a consent but were not enrolled (with analysis of EHR implementation of a Best Practice Alert). We summarized data from the EHR by eligibility, provider encounters, and alerts, and generated CONSORT diagrams and tables to trace the enrollment pathway. We supplemented quantitative findings with a thematic analysis of semi-structured individual interviews with eligible patients (n = 10) and study providers (n = 4) to identify systematic barriers to recruitment and enrollment of eligible patients. Results/Anticipated Results: CONSORT diagram analysis found that 24% of potentially eligible patients (268 of 1130) had an eligible study encounter but were not enrolled. Additionally, BPAs were not triggering for some eligible patients. Interviews revealed that study providers wanted more detailed information about which study arm their patient would be assigned to, and about next steps after enrollment, especially relating to additional lab tests and follow-up care needed. Patient interviews suggested that patients often did not remember completing the consent process and felt overwhelmed with appointments and information; patients expected providers to actively bring up research opportunities during appointments. Discussion/Significance of Impact: While pragmatic EHR-embedded trials are often characterized as lower-burden, these trials still require active engagement by providers, as well as ongoing attention from both research and informatics teams to ensure that EHR-embedded processes are functioning as designed, and that they are effective in recruiting study participants.

Objectives/Goals: Now in its 10th year, Research Jam, Indiana CTSI’s Patient Engagement Core, has proven to be an effective approach to patient engagement, foregoing generalized CABs for study-specific groups, personally invested in the research. Here we share our methods to benefit CTSAs seeking deeper engagement. Methods/Study Population: Research Jam is unique from patient engagement efforts in other CTSAs in almost every aspect. The composition and background of our team, the methodology to our practice and often our outcomes. This poster will use 10 years of projects, publications, and participant evaluations to explore our approach both quantitatively and qualitatively to see how it has contributed to the Indiana CTSI’s commitment to community engagement. Results/Anticipated Results: We will present quantitative data from 10 years of participant evaluations illustrating that Research Jams are received positively by those that attend, and qualitative analysis of the evaluations around themes of feeling heard, valued, and integral in research. By finding themes in evaluations across 10 years of studies, with different topics, different population types, and different investigators, we can speculate on what aspects of our approach appeal to the community, contribute to their positivity toward research and researchers, and could be reproducible in other CTSAs. Discussion/Significance of Impact: We use “blueprint” instead of “toolkit” when describing our process, as we hope others can see this not as a tool to fix a problem, but a plan to be iterated on in agreement with the community that will benefit from it. The relationships researchers need to build with communities are not cookie cutter neighborhoods, but rich, colorful and vibrant ones.

Objectives/Goals: The objective of the Clinical and Translational Science Awards (CTSA) Program Collaborative and Innovative Acceleration (CCIA) Award Initiative is to support synergistic collaborations to develop, demonstrate, and sustainably implement innovative solutions across and beyond the CTSA Consortium. Methods/Study Population: All CCIA awards between 2016 and 2022 were reviewed and analyzed by Fiscal year, activity code, and research area using NIH analytical tools and platforms. Subject matter experts categorized each award by research topics, study populations, stage of translational science, and innovation type. The number and type of collaborating organizations were noted and major accomplishments and expected outcomes were summarized. Results/Anticipated Results: Between FY2016 and FY2022, NCATS funded 37 U01 and 18 R21 CCIA awards including >90 different public and private partnering organizations. CCIA awards spanned all stages of translation including preclinical (26%), clinical (36%), implementation (31%), and public health research (7%). Of the 55 CCIA awards, 31% focused on urgent public health needs and 25% were designed to address health disparities. Broadly, types of innovations included: Data science-related projects (18%), clinical care innovations (15%), biomarker or clinical outcome assessments (13%), digital health solutions (11%), therapeutic development (11%), therapeutic discovery (9%), education and training (7%), diagnostic tools (5%), software tools (5%), or tools for clinical research (5%). In total, >735 publications cited CCIA awards. Discussion/Significance of Impact: For >8 years, the CCIAs have brought together researchers from diverse scientific disciplines across the nation to speed the development of new health solutions with broad impact. Advancements in genomic screening, for example, have led to policy changes while new delivery approaches have improved the quality of care for underserved populations.

Objectives/Goals: The Standards for Reporting Implementation Studies (StaRI) are the Enhancing the Quality and Transparency of Health Research (EQUATOR) Network 27-item checklist for Implementation Science. This study quantifies StaRI adherence among self-defined Implementation Science studies in published Learning Health Systems (LHS) research. Methods/Study Population: A medical librarian-designed a search strategy identified original Implementation Science research published in one of the top 20 Implementation Science journals between 2017 and 2021. Inclusion criteria included studies or protocols describing the implementation of any intervention in healthcare settings. Exclusion criteria included concept papers, non-implementation research, or editorials. Full-text documents were reviewed by two investigators to abstract and judge StaRI implementation and intervention adherence, partial adherence, or non-adherence. Results/Anticipated Results: A total of 330 documents were screened, 97 met inclusion criteria, and 47 were abstracted including 30 research studies and 17 protocols. Adherence to individual StaRI reporting items ranged from 13% to 100%. Most StaRI items were reported in >60% of manuscripts and protocols. The lowest adherence in research studies was noted around economic evaluation reporting for implementation (16%) or intervention (13%) strategies, harms (13%), contextual changes (30%), or fidelity of either the intervention (34%) or implementation (53%) approach. Subgroup analyses were infrequently contemplated or reported (43%). In protocols, the implications of the implementation strategy (41%) or intervention approach (47%) were not commonly reported. Discussion/Significance of Impact: When leveraging implementation science to report reproducible and sustainable practice change initiatives, LHS researchers will need to include assessments of economics, harms, context, and fidelity in order to attain higher levels of adherence to EQUATOR’s StaRI checklist.

Objectives/Goals: The objectives of the study were to evaluate end-user feedback regarding usefulness and compliance with the revised DoD PRA-CR. The PRA-CR utilizes symptom-guided management strategies to advance service members with acute concussion through 6 stages of gradually increased activity prior to their Return to Duty (RTD). Methods/Study Population: Clinical providers previously trained on the PRA-CR were invited via email to participate in an online survey-based study to examine their opinions and utilization of the revised PRA-CR. Participants who responded to the initial email invitation were provided an electronic Microsoft Forms based survey. Of the 83 total responders, 36 met inclusion criteria and advanced to the end-user survey. Six items were designed to assess inclusion–exclusion criteria (i.e., credentialed medical provider trained in the PRA-CR with experience treating concussion over the previous 2 years). Four items gauging utilization required yes/no responses; 20 opinion items on a 7-point Likert scale ranged from strongly disagree to strongly agree; 5 explanatory items were multi-select; and 1 item allowed free text responses. Results/Anticipated Results: Overall, 87% of respondents who had used the revised CR indicated that it helped them treat patients with acute concussion and 73% rated, “ease of use” favorably. Of the newly added elements to the CR, utilization of the Patient Leadership Guide (PLG) was the highest at 78%, with the majority of the providers rating the PLG as useful in communicating with patients and command. In contrast, only 35% of participants reported using the Physical RTD screening section and 22% indicated using the Cognitive RTD screening tool. Those not utilizing the Physical screening identified a lack of support staff (67%) or setting barriers (47%) as the primary reasons. Those not utilizing the Cognitive RTD screening tool identified multiple barriers to use including availability (72%), inexperience (39%), and baseline data access (33%). Discussion/Significance of Impact: This study sought end-user (provider) feedback regarding the revised PRA-CR’s usability and utility, in addition to their confidence in the tool itself. Overall results were generally positive, except for the updated Physical RTD and newly introduced Cognitive RTD screenings.

Objectives/Goals: The objective of this study is to explore strategies for AI-physician collaboration in diagnosing acute respiratory distress syndrome (ARDS) using chest X-rays. By comparing the diagnostic accuracy of different AI deployment methods, the study aims to identify optimal strategies that leverage both AI and physician expertise to improve outcomes. Methods/Study Population: The study analyzed 414 frontal chest X-rays from 115 patients hospitalized between August 15 and October 2, 2017, at the University of Michigan. Each X-ray was reviewed by six physicians for ARDS presence and diagnostic confidence. We developed a deep learning AI model for detecting ARDS and explored the strengths, weaknesses, and blind spots of both physicians and AI systems to inform optimal system deployment. We then investigated several AI-physician collaboration strategies, including: 1) AI-aided physician: physicians interpret chest X-rays first and defer to the AI model if uncertain, 2) physician-aided AI: the AI model interprets chest X-rays first and defers to a physician if uncertain, and 3) AI model and physician interpreting chest X-rays separately and then averaging their interpretations. Results/Anticipated Results: While the AI model (84.7% accuracy) had higher accuracy than physicians (80.8%), we found evidence that AI and physician expertise are complementary. When physicians lacked confidence in a chest X-ray’s interpretation, the AI model had higher accuracy. Conversely, in cases of AI uncertainty, physicians were more accurate. The AI excelled with easier cases, while physicians were better with difficult cases, defined as those where at least two physicians disagreed with the majority label. Collaboration strategies tested include AI-aided physician (82.4%), physician-aided AI (86.9%), and averaging interpretations (86%). The physician-aided AI approach had the highest accuracy, could off-load the human expert workload on the reading of up to 79% chest X-rays, allowing physicians to focus on challenging cases. Discussion/Significance of Impact: This study shows AI and physicians complement each other in ARDS diagnosis, improving accuracy when combined. A physician-aided AI strategy, where the AI defers to physicians when uncertain, proved most effective. Implementing AI-physician collaborations in clinical settings could enhance ARDS care, especially in low-resource environments.

Objectives/Goals: The objective of this research is to characterize the role of anemia and its treatment with iron in the development of oxygen-induced retinopathy and to identify possible mechanisms of action for future investigation. Methods/Study Population: Newborn rats were exposed to cyclic hyperoxia (50%) hypoxia (10%) for 14 days to induce oxygen-induced retinopathy (OIR). Half of the pups were phlebotomized to induce anemia. Half of anemic and control pups received high-dose iron, while the other half received standard, low-dose iron. Retinas were dissected at postnatal day 20, and vascular outcomes were measured to determine phenotype. Comparisons will be made by two-way ANOVA. Additional future studies are planned: 1) electroretinogram to measure retinal function, 2) measure of retinal hypoxia and angiogenic protein levels to evaluate the effect on hypoxia-sensitive angiogenesis, 3) serum and retinal iron measures to determine iron status, 4) flow cytometry and cytokine array to evaluate immune cell response, and 5) exploratory single-cell RNA sequencing. Results/Anticipated Results: Preliminary vascular outcomes are pending. We anticipate that retinas of anemic pups treated with low-dose (standard of care) iron will exhibit iron deficiency and hypoxia and reduced severity of OIR. Retinas of anemic and non-anemic pups treated with high-dose iron will exhibit iron excess and increased severity of OIR. However, retinal function will be reduced in anemia and rescued with treatment with high-dose iron. We further anticipate that anemia and iron-deficiency will down-regulate overall immune cell fractions, which will correlate with reduced cytokine and chemokine levels, suggesting a reduced immune response. Finally, we expect vascular endothelial cells and pericytes in anemic pup retinas to show increased angiogenic gene expression and neurons to show signs of impaired development. Discussion/Significance of Impact: We expect that anemia has a differential impact on retinal vascular anatomy and neuronal function. While retinopathy severity may be improved, overall retinal function will be dampened. These findings and the mechanistic work of this study will generate targets for intervention to preserve improved retinopathy outcomes but rescue retinal function.

Objectives/Goals: Our main objective was to compare 5-year survival and organ function between patients with sickle cell disease (SCD) who underwent hematopoietic cell transplant (HCT) and those who did not undergo HCT. We hypothesized that organ function would be improved in those with SCD who underwent HCT when compared to those who remained on standard therapy. Methods/Study Population: This IRB-approved, retrospective study includes patients with SCD treated at Children’s Healthcare of Atlanta. Cases underwent HCT between 2010 and 2016. They were randomly matched with 2 patients with SCD who did not undergo HCT. Match criteria included age, sex, disease genotype, and disease severity, which was determined by the number of hospitalizations in the 5 years pre-HCT, prior intensive care unit admission, and prior chronic transfusion therapy. Data extracted included SCD treatment, hospitalizations, emergency department visits, and organ function pre-HCT and 1-, 2-, 3-, and 5-years post-HCT. Organ-specific outcomes and overall survival were compared between the two groups using cumulative incidence curves and Kaplan–Meier analyses. Normal FEV1 and FVC in this analysis were >80% predicted. Results/Anticipated Results: Thirty-seven cases who had undergone HCT were matched with 74 controls who continued with standard medical therapy. The median age was 8 years for both groups and 59% were females. The median disease severity score was 2 in both groups. At baseline, 70.3% of the HCT group completed pulmonary function tests (PFTs) compared to 35.1% of the non-HCT group. Of these, 73% in both groups had a normal FEV1. In terms of FVC, 57.7% of HCT patients and 76.9% of non-HCT patients had a normal FVC pre-HCT. At 5 years post-HCT, 56.8% of the HCT group had PFTs completed compared to 21.6% of the non-HCT group. Among these, 85.7% in the HCT group had a normal FEV1 compared to 75% in the non-HCT group, while 90.6% had a normal FVC in the HCT group compared to 75% in the non-HCT group. Two of 37 in the HCT group and 1 of 74 in the non-HCT group died (p = 0.21). Discussion/Significance of Impact: Our data suggest that post-HCT, the proportion of patients falling in the normal range for FEV1 and FVC increases. This increase is not seen in the non-HCT group, indicating that HCT may improve this organ function. There was no difference in survival between the groups, indicating the risk of HCT mortality may not be greater than the risk of living with SCD.

Objectives/Goals: Dravet syndrome is a developmental and epileptic encephalopathy associated with refractory seizures and a high risk of sudden unexpected death in epilepsy. A pathogenic biallelic variant in SCN1B, SCN1B-p.R98C, was identified in three patients with Dravet syndrome. Here we investigate SCN1B-p.R98C on neuronal function in vivo. Methods/Study Population: Scn1b-p.R98C mice were previously generated using CRISPR-Cas9 gene editing. Homozygous animals exhibit increased susceptibility to hyperthermia induced seizures at postnatal day (P) 15, 100% expression of spontaneous generalized seizures by P30, and ~20% undergo SUDEP by approximately P60. Here we examined the neuronal phenotype of P17–28 male and female Scn1b-p.R89C mice. We used whole-cell patch clamp electrophysiology approaches to measure effects of the variant on passive membrane properties, intrinsic excitability, and single action potential properties of parvalbumin positive (PV+) interneurons and pyramidal neurons in layers 5/6 of the somatosensory cortex and CA1 region of the hippocampus. Wild-type littermates were used as controls. Results/Anticipated Results: Our results show no differences between genotypes in any measure for somatosensory cortical PV+ interneurons or pyramidal neurons. In the CA1 region of the hippocampus, we found no differences for any measure in PV+ interneurons. In contrast, CA1 pyramidal neurons were hyperexcitable, however, with no changes in passive membrane properties or single action potential properties. Discussion/Significance of Impact: