Objectives/Goals: Urinary tract infections (UTIs) cause significant morbidity, and many patients require multiple courses of antibiotics increasing the risk of antibiotic resistance. We determined the prevalence of urinary antibiotic heteroresistance (HR), which has been associated with treatment failures in vivo, to three first-line antibiotics for UTIs. Methods/Study Population: Clinical urine Escherichia coli isolates from patients in metropolitan Atlanta, Georgia in August 2023 were collected as part of public health surveillance performed by the CDC-funded, Georgia Emerging Infections Program (EIP). Only the first E. coli isolate collected for each patient was included in this study. Antibiotic susceptibility was determined through medical record review. HR to nitrofurantoin, trimethoprim-sulfamethoxazole, and fosfomycin was determined by population analysis profiling (PAP), where broth dilutions of E. coli were plated on increasing concentrations of the antibiotic. HR was defined as survival of >1 in 106 cfu but fewer than 50% survival at 1X antibiotic breakpoint (bp), resistant as > 50% survival at 1X bp and susceptible as survival of Results/Anticipated Results: Among 355 patients, 21 (5.9%) were resistant or intermediate to nitrofurantoin and 92 (26%) were resistant to trimethoprim-sulfamethoxazole. Antibiotic susceptibility data were missing from 5(1.4%) and 11(3%) of isolates for nitrofurantoin and trimethoprim-sulfamethoxazole, respectively. Susceptibility testing was not routinely performed nor reported for fosfomycin, thus excluded. PAP revealed that of the total 355 isolates, 3(0.84%) were heteroresistant to nitrofurantoin, 17(4.8%) were heteroresistant to trimethoprim-sulfamethoxazole, and 27(7.6%) were heteroresistant to fosfomycin. Of the isolates found to be susceptible by standard testing, 1(0.3%) and 9(3.6%) were heteroresistant to nitrofurantoin and trimethoprim-sulfamethoxazole by PAP, respectively. Discussion/Significance of Impact: Despite low rates of HR to nitrofurantoin and trimethoprim-sulfamethoxazole (0.84%, 4.8%), HR to fosfomycin was more frequent (7.6%). Given that susceptibility is not generally performed for fosfomycin, this could have implications for including fosfomycin as a first-line treatment for E. coli UTIs.

Objectives/Goals: The overarching goal of the GRAMM study is to address racial health disparities by increasing access to genetic counseling and testing among the Black community. Specific objectives are to determine patient acceptability of risk assessment at time of mammography and to evaluate subsequent access to and uptake of genetic counseling and testing. Methods/Study Population: All women presenting for screening mammography at the Ypsilanti Health Center under the University of Michigan were invited to enroll. After providing written informed consent, study participants entered family cancer and personal health information in the InheRET software tool which links to the National Comprehensive Cancer Network genetic testing guidelines. Upon completion, each participant was informed immediately if they did or did not meet the criteria to meet with a genetic counselor. For those who met the criteria, referral to genetic counseling was provided. All enrollees were invited to complete a post-assessment survey on acceptability of the service and genetics knowledge. Patients will be followed over time for completion of genetic counseling and testing. The study was approved by the Umich IRB. Results/Anticipated Results: As of 10/18/24, a total of 53 were enrolled, with 52 eligible for the study and were between the ages of 30–75 years. 51 identified as women and 1 identified as nonbinary. 21 (40.4%) identified as Black, 30 (57.7%) identified as White, 2 (3.8%) identified as Hispanic, and 1 (1.9%) identified as mixed race. Of the total enrolled, 25 (48.1%) met criteria to meet with a genetic counselor. Twelve (23.1%) have been scheduled to with meet with a genetic counselor and 2 (3.8%) of this group completed their appointment, but did not pursue genetic testing. 28 (53.8%) completed the survey and reported that they were satisfied with the service. Of the 16 people who screened positive and completed the survey, all 16 (100%) stated that they intended on proceeding with testing. Our study is still ongoing. Discussion/Significance of Impact: While this model has demonstrated acceptability so far, there are still possible barriers to genetic counseling and testing after the referral has been provided that need to be explored. However, this approach could provide a novel framework for combining risk assessment with screening mammography for all women nationwide.

Objectives/Goals: Our team has developed a 24-well donor-derived skeletal muscle microphysiological system (MPS) to study signaling pathways associated with a variety of muscle diseases. 3D muscle will be utilized to evaluate pharmacologic interventions for these muscle conditions to improve both muscle mass and function. Methods/Study Population: In this study, muscle MPS were formed from healthy young female and male subjects. 3D muscle underwent a 21-day differentiation with an electrical stimulation (e-stim) regimen twice daily beginning on Day 14. Functional assessments in permeabilized fibers of both sexes included isometric and isotonic calcium-induced contractions, allowing for the characterization of force-pCa (-log[Ca2+]), force-velocity and force-power relationships. Samples from Day 17 and Day 21 will be assessed for pro-growth protein signaling via western blotting and a subset of samples will be analyzed by histology and microscopy for fiber type and size. Finally, culture media pre- and post-terminal e-stim on Day 21 will be collected for extracellular vesicle (EV) isolation and EVs will be assessed by standard proteomics analysis. Results/Anticipated Results: Permeabilized fibers from both sexes reproduced the well-established sigmoidal force-pCa and the curvilinear force-velocity and force-power relationships reported in native striated muscle. Maximum specific force and force-pCa relationship were not different between sexes. Isotonic contractile measurements revealed that these male and female fibers also exhibit similar force-velocity and force-power relationships. We anticipate that 3D muscles from day 17 compared to day 21 will exhibit higher levels of pro-growth protein signaling due to e-stim application and no differences in fiber type or size between sexes. Additionally, we expect that EV quantity will depend upon 3D muscle maturity and presence of e-stim. Discussion/Significance of Impact: This study demonstrates the similarities of functional characteristics and exercise (or e-stim) adaptation between native human skeletal muscle and 3D bioengineered skeletal muscle. Ultimately, this data further validates the muscle MPS system to study muscle diseases and to enhance the translation of therapeutics to clinical settings.

Objectives/Goals: In alignment with the Clinical and Translational Science Awards (CTSA) UM1, continuous quality improvement (CQI) needs to be integrated into the elements and hub evaluation. As a first step to operationalizing this process, the University of Cincinnati hub (CCTST) evaluation team developed a systematic approach to prioritizing and sequencing tasks for aligning evaluation methods with CQI. Methods/Study Population: A scoring sheet was developed to provide a framework for prioritizing and developing the sequence and timeline for supporting elements in aligning their evaluation methods with CQI. The scoring sheet assessed: 1) impact – defined as the results from completing the task and implementing enhancements; 2) effort – defined as the amount of resources (time, personnel, and materials) needed to complete the tasks; 3) reach – defined as number of individuals (e.g., CTSA employees, members, researchers, trainees, and community members) impacted by the tasks or products of the tasks; 4) urgency – defined as a task that is time-sensitive due to a deadline and has a clear consequence if not completed on time. Each component was assessed using a 3-point scale (e.g., minimal, moderate, and high). Results/Anticipated Results: In Fall 2024, the CCTST evaluation team met with respective elements to collect data on their: 1) need from the evaluation team to support aligning their evaluation methods with CQI, 2) challenges and barriers to improving evaluation and aligning with CQI methods, 3) number of hours per month available to support improving evaluation methods, and 4) current resources to dedicated to conducting an evaluation. Next, the evaluation team will transcribe the data from the meetings and code the data into the scoring sheet for each element. The scoring sheet is anticipated to produce a score that will be used to develop the sequence, timeline, and initial tasks for supporting elements in improving evaluation methods and aligning with CQI over the first year of the UM1. Discussion/Significance of Impact: CTSA hubs’ evaluation teams operating at full capacity may encounter barriers to implementing CQI efforts. This systematic approach – assessing the impact, effort, reach, and urgency to sequence evaluation and CQI alignment – can support evaluation teams by ensuring a balanced workload and optimizing operations for quality improvement.

Objectives/Goals: We describe the prevalence of individuals with household exposure to SARS-CoV-2, who subsequently report symptoms consistent with COVID-19, while having PCR results persistently negative for SARS-CoV-2 (S[+]/P[-]). We assess whether paired serology can assist in identifying the true infection status of such individuals. Methods/Study Population: In a multicenter household transmission study, index patients with SARS-CoV-2 were identified and enrolled together with their household contacts within 1 week of index’s illness onset. For 10 consecutive days, enrolled individuals provided daily symptom diaries and nasal specimens for polymerase chain reaction (PCR). Contacts were categorized into 4 groups based on presence of symptoms (S[+/-]) and PCR positivity (P[+/-]). Acute and convalescent blood specimens from these individuals (30 days apart) were subjected to quantitative serologic analysis for SARS-CoV-2 anti-nucleocapsid, spike, and receptor-binding domain antibodies. The antibody change in S[+]/P[-] individuals was assessed by thresholds derived from receiver operating characteristic (ROC) analysis of S[+]/P[+] (infected) versusS[-]/P[-] (uninfected). Results/Anticipated Results: Among 1,433 contacts, 67% had ≥1 SARS-CoV-2 PCR[+] result, while 33% remained PCR[-]. Among the latter, 55% (n = 263) reported symptoms for at least 1 day, most commonly congestion (63%), fatigue (63%), headache (62%), cough (59%), and sore throat (50%). A history of both previous infection and vaccination was present in 37% of S[+]/P[-] individuals, 38% of S[-]/P[-], and 21% of S[+]/P[+] (P<0.05). Vaccination alone was present in 37%, 41%, and 52%, respectively. ROC analyses of paired serologic testing of S[+]/P[+] (n = 354) vs. S[-]/P[-] (n = 103) individuals found anti-nucleocapsid data had the highest area under the curve (0.87). Based on the 30-day antibody change, 6.9% of S[+]/P[-] individuals demonstrated an increased convalescent antibody signal, although a similar seroresponse in 7.8% of the S[-]/P[-] group was observed. Discussion/Significance of Impact: Reporting respiratory symptoms was common among household contacts with persistent PCR[-] results. Paired serology analyses found similar seroresponses between S[+]/P[-] and S[-]/P[-] individuals. The symptomatic-but-PCR-negative phenomenon, while frequent, is unlikely attributable to true SARS-CoV-2 infections that go missed by PCR.

Objectives/Goals: Gait performance (GP) and global cognitive functions (GCF) are both critical for maintaining independence and quality of life in senior adults. Recent studies have explored the potential link between GP and GCF, encompassing executive functions. Methods/Study Population: PRISMA guidelines will govern this systematic review. This systematic review synthesizes published research from 2000 to 2024, including peer-reviewed articles, pilot studies, and randomized controlled trials, to examine the relationship between GP (how a person walks and stands) and GCP in older adults. The exclusion criteria will be based on studies focused on physical activities unrelated to balance, meta-analysis, and systematic reviews and those published in languages other than English or Spanish. Results/Anticipated Results: Our preliminary data indicate that gait, or walking speed, is significantly correlated with GCP in older adults, with slower walking associated with poorer global cognition. Specifically, gait speed during dual-task walking shows a strong correlation with working memory (p < .001) and processing speed (p < .05) in individuals aged over 60 years. Gender differences were observed, with women over 80 walking slower than men over 85 years, who walked faster, and women exhibited poorer global cognition than men. Discussion/Significance of Impact: Overall, a gait slowing between 0.68 and 1.1 m per second could predict a marker for the risk of developing dementia, indicating that monitoring gait speed in older adults may provide early warning signs, allowing for timely interventions. Enhancing GP and GCF can improve the quality of life and independence in older adults. Acknowledgments: Research supported by NIH: Award Number HCTRECD R25MD007607 from the NIMHD.

Objectives/Goals: Explore the association of gestational age of LPIs and neurodevelopment delay in children exposed to the neonatal intensive care unit (NICU). Compare between groups if exposure to NICU affects neurodevelopment in children who were born between 34 and 37 weeks of gestation. Methods/Study Population: This is a cross-sectional study design to study the association of gestational age and neurodevelopmental delays in a birth cohort in Puerto Rico of children exposed to the NICU. Their neurodevelopment will be measured with the Bayley III. Statistical analysis will be performed using IBM SPSS Statistics 27.0. Descriptive statistics will be used, and normality distributions among all continuous variables, frequency distribution for categorical variables. We will recruit 30 infants between 18–24 months of age that were born between 34–37 weeks of gestation. Infant will be divided with regard to their exposure to NICU experience. Results/Anticipated Results: We anticipate identifying neurodevelopment delays among children born prematurely between 34 to 37 weeks of gestation. We anticipate that our controlled group will have better outcomes when compared to the controlled expose group. We also expect that gestational age impacts adversely neurodevelopment in children who were born between 34 and 37 weeks of gestation. Discussion/Significance of Impact: Approximately 84% preterm birth are considered LPIs. Prematurity is described as a chronic condition; adverse long-term neurodevelopment consequences. Our study promotes early detection and interventions that can reduce the consequences of the neurodevelopment delays in LPIs.

Objectives/Goals: Some children with asthma benefit from care from asthma specialists: allergist/immunologists or pulmonologists. However, a limited supply of specialists poses access problems for some children. Our objective was to describe the state of specialist care for pediatric asthma in Maine. Methods/Study Population: Using Maine all-payer claims data for April 2018–April 2021, we identified children with asthma meeting criteria for subspecialist referral based on NIH Expert Panel Report criteria. We determined which of these children had encounters with an allergist-immunologist or pediatric pulmonologist during the study period and performed logistic regressions to determine what demographic traits were associated with receipt of care. Results/Anticipated Results: Among children meeting criteria for specialists referral, 33% had an encounter with an asthma specialist. Specialist encounters were associated with younger age (p<0.001) and male sex (p = 0.003). Encounters were negatively associated with Medicaid insurance (versus commercial, p<0.001) and living in more rural areas (RUCA codes 4–10, p<0.001), lower childhood opportunity index (high or less, <0.001), and longer travel time to pediatric asthma specialists in Maine (<0.001). Discussion/Significance of Impact: Specialist care for children with complex or severe asthma is an evidence-based practice that is not equally accessible to all children in Maine. This may restrict their access to specific asthma therapies such as allergic desensitization and monoclonal antibodies.

Objectives/Goals: The overall goal of this project is to determine bacterial transcriptional signatures from clinical sputum and assess their potential to monitor treatment response and predict the outcome of drug therapy in patients with tuberculosis (TB). Methods/Study Population: We are developing a novel transcript capture sequencing (TC-Seq) approach to sequence the mRNA of Mycobacterium tuberculosis (Mtb) and analyze transcriptomes from clinical samples containing minimal amounts of bacterial RNA. This protocol generates single-stranded biotinylated probes from Mtb DNA. Probes are hybridized to and allow enrichment of Mtb-specific mRNA within next-generation RNA sequencing libraries. We will apply TC-Seq to sputum samples collected throughout an 18-month Phase II clinical trial investigating response to TB treatment to compare the transcriptome of Mtb between patients whose treatment results in cure or relapse. Results/Anticipated Results: We have refined a technique to generate biotinylated probes starting from DNA of lab grown Mtb. This protocol achieves robust and unbiased sampling of the Mtb transcriptome from mixed samples containing both human and Mtb RNA. Preliminary sequencing of clinical sputum collected pretreatment has generated 1–4 million Mtb-specific reads, a sequencing depth that allows examination of the entire bacterial transcriptome. We will measure differential gene expression before and during treatment as well as between cure and relapse cases. These results will allow us to characterize bacterial response to treatment and identify bacterial markers that correlate with relapse. Discussion/Significance of Impact: Understanding Mtb activity during treatment will offer new ways to assess the efficacy of different treatment regimens. Crucially, identifying clear bacterial markers that demarcate a cure or relapse outcome will have a significant impact on determining patient eligibility for shorter drug therapy.

Objectives/Goals: Present a framework for hosting Community Grand Rounds, where community and academic partners showcase completed community-engaged research (CEnR) projects. This highlights innovative dissemination methods, engages diverse audiences, elicits community responses, and advances the translational science of CEnR. Methods/Study Population: Our approach involves planning and outreach to collaborate with promotion of academic and community engaged grantees to develop community dissemination events that translate the science of CE into accessible, relatable, culturally relevant formats for diverse audiences. These events incorporate interactive presentations that encourage active participation and feedback from attendees. Following each event, an evaluation is completed to assess community impact. Key strategies for hosting, facilitating, and utilizing diverse marketing to ensure that events are tailored to culturally diverse community groups, including regional implementation when practical. This collaborative approach meets a critical need and strengthens the bond between researchers and the communities they aim to serve. Results/Anticipated Results: These events create a feedback loop between the community and academic researchers. It was not just about telling people what was found. We created opportunities for community members and academics to build trust, give us feedback, ask questions, and discuss how findings could be practically applied. By presenting the findings in an accessible way within the community, community members are more informed and empowered to make decisions or advocate for changes in their own lives based on the research. Academics also benefited from community feedback, which provided new insights to help refine future research questions and methods. The goal is for shared conversation and understanding between community members and academics to inspire real-world applications and policy change directly informed by the research. Discussion/Significance of Impact: Community Grand Rounds are one dissemination strategy to leverage community–academic collaboration to present tailored research, fostering engagement, understanding, and action between researchers and community members. This approach effectively enhances the translational science of CEnR by involving and benefiting the community.

Objectives/Goals: The goal of the Accelerated Staff Assist Program (ASAP) is to help fill staffing gaps in clinical and translational research. The program offers centralized staff to fill short-term needs including project startup prior to hiring, project implementation, parental leave, and Spanish-language research support. Methods/Study Population: Promotion of ASAP in the first year was targeted to Penn State College of Medicine faculty to determine demand. Current outreach includes promotion to all Penn State campuses through PSCTSI’s newsletter, seminars, and presentations. Consultation intake data includes a project summary, staffing needs, and funding information. A project agreement, charter, and budget are drafted and agreed to by the principal investigator before work begins. Assessments occur at 6 and 12 months to assess satisfaction of the consultation process and quality of staff support, Net Promoter Score, and qualitative feedback about the program. Results/Anticipated Results: In its first year, ASAP conducted 26 consultations and supported 10 projects. Approximately 75% of staff effort has been recovered through agreements during the most recent 6 months. In anticipation of project delays, we found that booking FTE up to 115% results in a consistent recovery of 75% FTE. Consultee needs include short-term, partial effort support (76% of requests), project start-up (12%), staff leave (4%), and other (8%). ASAP staff tasks include participant recruitment and retention, data collection and analysis in English and Spanish, and community engagement for recruitment. Program assessments with research teams are ongoing; preliminary qualitative and quantitative data are unanimously positive. Discussion/Significance of Impact: The ASAP program has demonstrated fiscal success and sustainability in its first year. Hiring of a well-qualified, bilingual staff person allowed for quick onboarding and project support within one month of hire. ASAP enables CTSI to fulfill our mission by providing critical support for research teams to meet objectives and achieve equity goals.

Objectives/Goals: The NIH Common Fund launched Community Partnerships to Advance Science for Society (ComPASS) to study ways to reduce health disparities by addressing underlying structural factors within communities. ComPASS was designed for community organizations to lead research that addresses community needs. Methods/Study Population: ComPASS awarded five health equity research hubs (Hubs) to provide specialized technical support to ComPASS research projects led by community organizations. Expertise provided by the Hubs to the community-led projects will focus on specific facets of community health, including nutrition access, health care access, and built environment. The Hubs support community-led piloting and testing of structural interventions within community settings by providing subject matter expertise in areas including structural and multilevel intervention study design and methods, implementation science, and community engagement. Results/Anticipated Results: The Hubs will provide expertise and support to the community-led research projects around one or more social determinants of health domains: health care access and quality, education access and quality, economic stability, social and community context, and neighborhood and built environment. The Hubs will help identify strategies for measuring health outcomes and assessing the effects of structural and contextual factors on intervention outcomes. We anticipate the ComPASS program will lead to a better understanding of how structural interventions that leverage multi-sectoral partnerships can advance health equity. Discussion/Significance of Impact: Through community-led research, ComPASS projects are implementing structural interventions to address social determinants and advance health equity. The technical scientific support rooted in health equity provided by the Hubs is essential to the success of these research projects.

Objectives/Goals: Longitudinal studies capture dynamic host–environment interactions crucial to microbiome research. Commonly used mixed-effects models can struggle with small clinical samples, like many endometrial cancer studies. We will compare them to simpler fixed-effects models, aiming to develop a new fixed-effects differential abundance alternative. Methods/Study Population: Starting with simulated data for linear models with a single treatment effect, we will incrementally increase complexity to reflect real-world microbiome data. To observe conditions of misspecification, simulated data of repeated measures will be generated using different distributions, while varying parameters and inter-class correlation. In each condition, linear fixed- and mixed-effects models will be fitted 1000 times to empirically determine p-value, type 1 error rate, and power. We will increase the verisimilitude of our approach by using endometrial cancer data from our lab as a real background, while adding simulated signals. This data will be used to assess extant differential abundance models against our novel fixed-effects model. Results/Anticipated Results: Our initial findings indicate that fixed-effects models maintain control over type 1 error rates in small samples, while mixed-effects models do not perform as reliably. We are now exploring the effects of confounders on type 1 error rates and power in linear models, while our next step will evaluate generalized linear models in a differential abundance context. We expect that the fixed-effects models will continue to be as reliable as mixed-effects models, while being less computationally complex. After establishing our theoretical basis, our novel model should perform comparably to mixed-effects models on pseudo-simulated endometrial cancer data and better at small sample sizes. We will finally apply our model to real endometrial cancer data to identify microbial markers of disease predisposition. Discussion/Significance of Impact: By characterizing and improving these statistical tools, we seek to increase the robustness and power of microbiome study results (especially at small sample sizes), ultimately supporting better-informed translational research decisions. This work underscores the importance of statistical methods in advancing microbiome research.

Objectives/Goals: To develop an informatics framework that will allow study of environmental effects on stillbirth at large scale (i.e., US-level) and leverage recent advances in machine learning and artificial intelligence to produce reproducible results that can be compared across multiple institutional settings. Methods/Study Population: Experimental exposure data are often available in “absolute time,” where a clinical event can be anchored using a timeline transformation. We associate each stillbirth event with a set of {ti…ti+1}L shapelets [1] associated with a location, L, and time intervals for the entire dataset. These shapeless are aggregated using a state-of-the-art shapelet classifier [2]. An autoencoder is used to reduce the dimensionality of the stillbirth classification and to cluster stillbirth events according to their corresponding exposure patterns. The stillbirth cluster can be analyzed for other nonexposure (i.e., genetic, SDoH, and demographics) factors, which may be enriched and/or depleted. Results/Anticipated Results: The framework we are developing leverages a shapelet-based approach to produce clusters of stillbirth events according to their corresponding exposure patterns. These clusters can be analyzed for depletion or enrichment of nonenvironmental factors. This analysis will inform how to formulate (or not) class models of exposure that can be more informative and have better predictive power than overall population models. Moreover, the finding of depletion and enrichment of physiological properties of the individuals may lead to novel physiological hypotheses to better understand the injury mechanisms that the environmental exposure profile produces. Discussion/Significance of Impact: Nearly 20,000 babies are stillborn in the USA each year [3]. Environmental exposures, usually studied as time averages over certain periods of time, have produced mixed results for stillbirth risk [4]. However, temporal profiles matter [1], and we argue that they can be assessed using shapelet technology.

Objectives/Goals: Intra-amniotic inflammation (IAI) is one of the leading causes of maternal/fetal morbidity globally, yet it is undiagnosed in 90–95% of cases. The purpose of this study was to assess precision and accuracy of a novel microsampling device for measuring cytokines in amniotic fluid (AF) to enable noninvasive evaluation of localized inflammation. Methods/Study Population: AF was obtained from discarded amniocentesis samples from 3 deidentified patients without known inflammation. Samples were spiked to 5 concentrations of interleukin-6 (IL-6) and tumor necrosis factor-a (TNF-a), then sampled using the Neoteryx Mitra volumetric microsampling device (20 µL). Dried/reconstituted samples were analyzed using the Luminex R&D Mag XL multiplex (IL-6, TNF-α) and compared to free-flowing AF. Inter-and intra-assay performances were evaluated across 5-runs in 3 standard cytokine concentrations. Recovery/linearity was assessed by a recovery curve and parameter estimates. Precision was assessed between-run and within-run using coefficients of variance (%CV). Accuracy was evaluated as agreement between microsampled and free-flowing AF using Bland–Altman plots. Results/Anticipated Results: TNF-α results were linear for all 3 patients across 5 concentrations. However, accuracy and recovery consistently failed (mean±SD percent recovery 176±21%). TNF-α results had acceptable precision with %CV within-run of 6.8–12.1% and across-runs of 14.0–15.9%. Microsampled TNF-α agreed with free-flowing sample: 16 of 18 (89%) were within 1SD of the mean difference (-24±36 pg/ml). IL-6 results were linear for 1/3 patients and had unacceptable accuracy and recovery for 13/15 samples (mean±SD percent recovery 764±469%). IL-6 results had acceptable precision (%CV within-run 6.2%, 7.5% and 11.2%; across-run .9–15.2%). Microsampled IL-6 agreed with free-flowing sample; 17 of 18 (94%) were within 1SD of the mean difference (-148±180pg/ml). Free-flowing vs. microsampling methods agreed best at low concentration. Discussion/Significance of Impact: This study provides preliminary support for noninvasive measurement of cytokines leveraging small amounts of leaking AF providing a promising alternative to amniocentesis and potential for assessing inflammation intrapartum. Clinical application requires development of reference ranges and association between cytokine levels and outcomes.

Objectives/Goals: Micro-credentialing programs provide a rapid solution to the growing shortage of clinical research professionals (CRPs) by expanding the applicant pool and improving training efficiency. This study evaluates the impact of a micro-credential course on CRP education and its potential to reduce staffing shortages. Methods/Study Population: To address the CRP staffing shortage, new and existing clinical research staff at the Rutgers Cancer Institute of New Jersey (CINJ) participated in a micro-credential badging course developed by NJ ACTS and Rutgers’ Master’s in Clinical Research Management Program. The course focused on key clinical research topics designed to equip participants with foundational knowledge. Post-completion, surveys were administered to both participants and CINJ management to assess the program’s effectiveness. The survey measured perceived knowledge gains, interest in further opportunities, and the program’s potential to alleviate staffing shortages. Survey results were analyzed to determine the program’s overall impact on CRP education and staffing challenges. Results/Anticipated Results: Survey results demonstrated significant knowledge gains among participants, with 85% reporting increased confidence in clinical research topics after completing the micro-credential course. Additionally, 70% of participants expressed interest in pursuing further opportunities in the field. CINJ management reported smoother onboarding processes and noted an improvement in job readiness among new hires. The CRC Badge has since been integrated into CINJ’s formal onboarding process. Overall, the micro-credential program contributed to expanding the CRP applicant pool, improving training efficiency, and offering a short-term solution to alleviate staffing shortages. Discussion/Significance of Impact: This research demonstrates the effectiveness of micro-credentialing in addressing the critical shortage of CRPs. By rapidly equipping staff with essential knowledge, the program broadens the applicant pool, enhances onboarding, and offers an immediate solution to workforce gaps.

Objectives/Goals: Translational researchers spend significant amounts of time finding available datasets and other research data resources for their purposes. Objectives of this program are develop and evaluate a multipronged approach to supporting researchers with existing data resources. Methods/Study Population: We established a dedicated service with expertise in data resources to increase awareness, understanding, and utilization of existing data resources. This program assists investigators and trainees discover appropriate data resources, formulate scientific problems in computable formats, advise on state-of-the-art data analytics, data management, build collaborations, mentor data users, and develop a service pipeline for streamlined data resource project management. This is accomplished through these essential functions: (1) Discover, catalog, document, and manage metadata resources, (2) train and present data resources to the research community, (3) provide individual consultations, and (4) explore and assess novel data resources. Results/Anticipated Results: In a phased approach, the data navigation program is performing outreach to the research community and integrating with existing data efforts on campus, presenting and demonstrating existing data resources, established a consultation service, and building core competencies into long-term usage and navigation of resources across campus. Evaluating the program monthly has shown an increase in various metrics for evaluating commitment and engagement including number of requests for access to data resource, consultations, publications and presentations, co-authorship, and proposals. Unawareness and inappropriate use of data resources leads to delays in performing research and potentially unnecessary duplications of efforts. Discussion/Significance of Impact: Our data navigation program has increased use of data resources in research. Next steps are to continue evaluation and further streamline informatics approaches to data discovery, abstraction, formulation, and analysis. Harmonized data resource programs are important translational science approach to foster the next generation of research.

Objectives/Goals: Our study team won a Public Health Answers to Speed Tractable Results (PHASTR) contract to conduct a target trial emulation to answer “Does metformin show a reduction of severe outcomes of COVID-19 or of Long COVID in the N3C Data Enclave?” We quickly delivered an answer due to productive technical and collaboration support in the N3C. Methods/Study Population: Our analytic plan was updated based on helpful feedback from the PHASTR program. We performed a trial emulation analysis using the N3C data, comparing adult new users of metformin to controls prescribed fluvoxamine, fluticasone, ivermectin, or montelukast. The composite outcome was Long COVID or Death (LC/D) within 180 days of COVID infection. We used entropy balancing to estimate the average treatment effect with a weighted log-linear model. Productivity was enhanced by reusing code workbooks and validated codesets from related N3C projects. The team of 4 (physician, informaticist, data programmer, and statistician) and key unpaid advisors spent 10 weeks developing and analyzing the data. Results/Anticipated Results: Totally, 9,660 patients were identified for analysis. After weighting, there were 248 in the metformin and control groups. In the metformin group, 4.0% developed LC/D vs. 8.5% in the control group, with an adjusted risk ratio (aRR) of 0.47 (95% CI 0.25 to 0.89). Results were consistent across subgroups and sensitivity analyses. The PHASTR contract structure helped produce high-quality results quickly by not only providing funding but also requiring a compressed timeline for a small team to focus on the study. The most time was spent on contract execution, enclave provisioning, and too many last-minute download requests. A project final report was submitted in March and a full manuscript was submitted in September. Discussion/Significance of Impact: The analysis was productive because the environment made reuse easy and supported rich collaborations among clinicians, informaticists, epidemiologists, statisticians, and data developers. Advice from PHASTR advisors (Axel) and N3C diabetes domain team members was also key to a faster completion.

Objectives/Goals: Personalized cancer therapy based on genomic testing is advancing patient care. Genomic alterations in fibroblast growth factor receptor (FGFR) predict response to FGFR inhibitors; however, the role of RNA expression and protein activation is not known. We propose to examine the phospho-proteomic signature in FGFR-altered cancers to identify new candidates for FGFR-targeted therapies. Methods/Study Population: In our preliminary study, we have curated a cohort of FGFR2 mutants (13 FGFR2-fusions and 4 FGFR2 point mutations) with known clinical outcomes to FGFR inhibitors and 8 FGFR2 wild-type (WT) cholangiocarcinoma tumor samples to investigate the phospho-proteomic fingerprint using a clinical grade reverse phase protein array (RPPA). RPPAs are high throughput quantitative antibody-based proteomics assays that can quantify hundreds of proteins in thousands of patient tissues providing a high degree of sensitivity through laser tumor microdissection (LCM). We have selected proteins in the FGFR signaling pathway including FGFR2, AKT, ERK1.2, STAT1/3, FRS2, and PLCg to define the range of phospho-proteomic signal between FGFR2 WT and mutant cancers. All samples will undergo evaluation with RNASeq for gene expression. Results/Anticipated Results: Our initial analysis defined the range of RNA expression of FGFR2 and pFGFR2 protein signal (Y653/654 and Y769) between FGFR2 WT and FGFR2 mutant samples. On average, the FGFR2 mutant cohort displayed higher FGFR2 RNA expression compared to the FGFR2 WT cohort. There is no apparent correlation between RNA expression and clinical response to FGFR-targeted therapy. However, in this small cohort, there is no significant difference in FGFR2 phosphorylation between FGFR2 WT and mutant cancers. RPPA analysis of FGFR downstream signaling proteins reveals a wide range of phosphorylation, but no significant difference between FGFR2 WT and mutant cancers. Discussion/Significance of Impact: These findings illustrate the complexities of FGFR signaling between FGFR2 WT and mutant cancers. These data suggest that tumors with genomically WT FGFR may display increased pFGFR2 and downstream signaling phospho-proteins. We propose a larger study of cholangiocarcinoma to evaluate evidence of FGFR pathway activation in WT tumors.

Objectives/Goals: Inclusive physical activity (PA) interventions could improve trans and gender diverse (TGD) adolescents’ PA levels, perceived social support from peers and adults, and mental health, but no stakeholder-informed interventions exist. We describe parents’ impressions of TGD adolescent children’s PA experiences and advice for a PA intervention. Methods/Study Population: We conducted individual Zoom interviews with parents of 13- to 20-year-old TGD adolescents (n = 15). All parents were recruited from a gender healthcare clinic. All children self-identified as TGD, an umbrella term including people who have a gender identity different from social expectations for their assigned sex. We asked questions regarding children’s current and historical participation in PA, parents’ perceptions of barriers to PA for their TGD children, and parents’ desired intervention components. After each interview, parent participants were compensated $100 for their time. We analyzed interview transcripts, focusing on insights to incorporate in an intervention. This study was approved by the University’s Institutional Review Board. Results/Anticipated Results: Parents shared rich histories of their children’s PA participation, compounding barriers to PA, numerous benefits of PA, and a range of preferences for program activities and inclusive practices (e.g., safety protocols, training for adult leaders). They also emphasized the need for TGD youth to build social connectedness through PA. Discussion/Significance of Impact: We gathered concrete advice from parents on creating a PA intervention, which we will use to build a social PA program that meets TGD adolescents’ needs. Addressing health disparities and improving PA, social support, and mental health among TGD adolescents will require such stakeholder input to improve upon existing PA and sports opportunities.