OBJECTIVES/GOALS: The overall objective of this study is to determine if medial arterial calcification (MAC) is an independent predictor of diabetes and to evaluate the relationship between MAC, periodontitis and Type II diabetes mellitus. METHODS/STUDY POPULATION: A retrospective case-control model analyzing radiographs for periodontitis and MAC to identify potential biomarkers for underlying systemic conditions, such as diabetes. Charts of patients attending UPENN School of Dental Medicine clinics between 2015 and 2022 were reviewed. Demographics, medical and dental history, diabetic status (identified by POC blood glucose level, fasting blood glucose and/or A1C), and medication history were documented amongst other variables. Patients aged 18 years or older with diabetes and having full mouth intraoral radiographs (FMX), panoramic radiographs and CBCTs were included. Patients with radiographs of poor quality were excluded. Multivariate analysis was used to determine possible associations between diabetes and periodontitis among persons with or without MAC. RESULTS/ANTICIPATED RESULTS: In our pilot study involving 28 participants, 53% of the population with moderate or severe periodontitis had MAC. By the Fisher’s Exact Test, there was an association, meaning those with more periodontal disease are more likely to have MAC (p=0.014). Sixty-three percent of patients with diabetes had MAC, while 19% of patients without evidence of diabetes also had MAC, (p=0.067). There was not enough evidence of association between diabetes and presence of MAC at this time, due to a small sample size, however there was a high prevalence of MAC among the diabetics. We hypothesized that periodontitis, a condition that shares many risk factors with diabetes would also be associated with incident MAC. Findings from this study will be key for the implementation of preventive screening protocols and referral systems. DISCUSSION/SIGNIFICANCE: Diabetes is on the rise and about half of diabetics are undiagnosed. CBCT imaging frequently used in dentistry can detect incidental findings such as MAC. This study has the potential of detecting statistically significant links between MAC, periodontitis and diabetes, hence serving as a sensitive radiographic biomarker for diabetes.

OBJECTIVES/GOALS: There is a need for high-quality and efficient translation of health technologies in the Ontario healthcare system. The goal of this project is to understand the decision-making processes of government expert groups developing recommendations for the system-level implementation of pharmacogenomic testing. METHODS/STUDY POPULATION: This prospective observational case study includes the Ontario Health Pharmacogenomics (PGx) Working Group focused on developing recommendations for a PGx testing implementation strategy in the province. Ontario Health is the government agency that oversees provincial healthcare planning and service delivery. Using qualitative ethnographic methods, we will observe and document the working group’s activities over a 10-month period. Data collection involves meeting recordings, correspondences, researcher field notes, decision-making processes, and group characteristics. Using descriptive statistics and inductive qualitative analyses, the data will be examined to build theory and frameworks for knowledge translation. RESULTS/ANTICIPATED RESULTS: The results will be presented through a case report, process maps, decision milestones, visualizations, and procedural recommendations for future expert groups. This study will contribute to the body of foundational knowledge about translational sciences and support the National Center for Advancing Translational Sciences’guiding principles. To enhance translational processes and train the future translational workforce, this research can be used for educational initiatives. In addition, the observed processes will inform a theory about how expert recommendations are developed in public healthcare systems. DISCUSSION/SIGNIFICANCE: This research addresses a current gap in understanding around translational processes, government decision-making, and the development of recommendations for the adoption, implementation, and dissemination of the novel health technologies transforming public healthcare in Canada.

OBJECTIVES/GOALS: The Fulkerson Home Food Inventory (HFI) is widely used to assess the home food environment, a key target of behavioral weight loss trials. However, no standardized report is available. We created publicly available procedures to automate and standardize HFI reporting, yielding a personalized report to enhance this measures clinical utility. METHODS/STUDY POPULATION: Parents in the TEENS adolescent behavioral weight loss trial complete the HFI at 0-, 2-, 4-, 8-, and 12m and receive personalized reports at each timepoint. In REDCap, participants identify foods available in their home. HFI syntax is applied to calculate the obesogenic home food availability score. Categories of foods found are identified, with specific guidance provided to enhance their home food environment. Prior to automation, procedures were time intensive and error prone. To address this, HFI data are exported into Excel by a PowerShell (v7.2) command-line script using Python (v3.10) with the REDCap API. Results are calculated with F# (v6.0) using Microsoft Excel Interop API and inserted into a report template with F# using the Microsoft Publisher Interop API. This process is repeated at each timepoint. RESULTS/ANTICIPATED RESULTS: The new automated procedures significantly reduce time to generate reports and enhance accuracy. Procedures yield a 2-page individualized report that includes the obesogenic home food environment score and identifies categories of healthy items found (e.g., fruits, vegetables, whole grains) as well as areas of improvement (e.g., high-fat dairy products, processed meats). Specific items found in each category are identified. The report identifies food found in the home (e.g., chicken nuggets) with suggested healthier substitutions (e.g., lean chicken breast). This syntax and commands will be made publicly available for use in the scientific and clinical community. DISCUSSION/SIGNIFICANCE: These publicly available procedures optimize, automate, and standardize reporting for the HFI. Procedures improve efficiency within large-scale clinical trials and yield a personalized report to enhance the clinical utility of this measure and empower participants to make informed decisions about their health behaviors.

OBJECTIVES/GOALS: eHealth programs centered on cognitive behavior therapy (CBT) can be supported by physical therapists to feasibly deliver psychologically-informed physical therapy (PIPT). This study assessed patient and physical therapist (PT) perspectives of adding a CBT-based eHealth program to physical therapy. METHODS/STUDY POPULATION: In our uncontrolled pilot study, PTs were trained in motivational interviewing (MI) to support patient engagement with a 7-module eHealth CBT-based pain self-management program that accompanied a course of PT. Interviews were conducted with a convenience sample of 13 patients with chronic back and/or neck pain and 9 PTs to evaluate experiences with the eHealth program, perceived benefits, barriers and facilitators to integration, and future recommendations for implementation from both perspectives. Interview data were recorded, transcribed, and analyzed using qualitative content analysis for core themes. RESULTS/ANTICIPATED RESULTS: Patients benefited from the eHealth program, especially relaxation (69% of respondents) and meditation/mindfulness (62%). Time and technology were barriers for eHealth engagement (23% and 15%). Most (92%) patients reported their therapist reviewed eHealth progress, although one-third of patients wanted more discussion and feedback on eHealth content. PTs felt the eHealth program was valuable (67%), user-friendly (56%), and took minimal effort to support (78%). Some (44%) therapists felt more confident after MI training to support patient engagement. PTs recommended offering patients tablets (i.e., iPads) for clinic or home use and additional therapist training in MI. DISCUSSION/SIGNIFICANCE: PTs and patients valued a clinician-supported eHealth program, suggesting an acceptable option for PIPT. Future implementation work aims to identify optimal therapist training in MI and additional strategies to overcome time or technology barriers.

OBJECTIVES/GOALS: Residents of environmental justice (EJ) communities experience significantly higher rates of negative health outcomes associated with poor air quality. Low-cost air sensors may supplement regulatory monitoring to better measure air pollution at local scales, but widespread application of this technology remains limited due to many challenges. METHODS/STUDY POPULATION: To address these obstacles, we designed a training program to equip community and academic research partners with the skills and knowledge to successfully apply low-cost sensors in community-engaged environmental health research. The R esearch Innovations using Sensor Technology in Environmental Justice Communities (RISE Communities) program was established through an NIEHS R25 award in 2022 and has three specific aims: 1) Foster community-academic partnerships through research education, training, and team development activities, 2) Provide technical training in the application of low-cost sensors for indoor, outdoor, and personal air monitoring in EJ communities, and 3) Establish a community of practice to address air quality in communities nationwide. RESULTS/ANTICIPATED RESULTS: We hosted our first cohort in August 2023, training five community-academic research teams in team collaboration, community-engaged research, and technical skills for collecting and analyzing data from PurpleAir sensors. Each team received 12 sensors to take to their home EJ communities to begin their projects. Community of practice development continues through interactive webinars and development of a web-based repository of training videos and discussion board posts. Evaluation data show high participant learning and satisfaction, with mean confidence scores improving on 6/8 metrics. Evaluation data also suggest several areas for improvement such as more time spent in teams for planning and additional opportunities for interaction within the cohort and with program instructors for problem-solving. DISCUSSION/SIGNIFICANCE: Effective training for team-based community-engaged research requires careful planning for team development and study implementation. Longitudinal training and support for the technical aspects of utilizing air sensors is also critical to team success. The RISE Communities program is actively recruiting for future training cohorts.

OBJECTIVES/SPECIFIC AIMS: Our objective was to examine serum E2 levels in dcSSc males in relation to disease characteristics (i.e autoanitbody profile and internal organ involvement) and its impact on survival. METHODS/STUDY POPULATION: We measured serum E2 levels in 83 dcSSc men >50 years old from the University of Pittsburgh Scleroderma Center and healthy controls of similar age. Using statistical modeling, we examined the associations between circulating E2 levels, internal organ involvement, autoantibody profiles, and survival. RESULTS/ANTICIPATED RESULTS: Male dcSSc patients had significantly higher serum E2 levels compared to healthy male controls and compared to dcSSc post-menopausal women of similar age. Male dcSSc patients with high serum E2 levels had significantly more heart involvement and worse survival. Using Cox regression modeling for risk of death, increasing serum E2 levels in anti-Scl-70 antibody positive dcSSc males were associated an increased risk of death. DISCUSSION/SIGNIFICANCE OF IMPACT: DcSSc male patients have higher levels of E2 compared to healthy controls and dcSSc postmenopausal women. Elevated serum E2 levels in dcSSc males >50 are associated with heart involvement and, if anti-Scl-70 antibody positive, worse survival. Our current study expands on our previous work, not only that that E2 exerts pro-fibrotic effects on skin, but also internal organ involvement, overall survival. These data suggest an important role of estrogen imbalance in SSc.

ABSTRACT IMPACT: This study deepens knowledge with respect to the associations between depression, cardiometabolic conditions, and accelerated aging with a clinically accessible marker in a population with disproportionate risk for comorbidity. OBJECTIVES/GOALS: The aim of this secondary analysis is to examine associations between DNA methylation age acceleration (DNAm AA) and depressive symptoms in African American women (AAW) considering the presence of cardiometabolic conditions (CMCs) including hypertension, diabetes, obesity. METHODS/STUDY POPULATION: Genomic and longitudinal clinical data (collected 2015-2020) from the Intergenerational Impact of Genetic and Psychosocial Factors on Blood Pressure Study (InterGEN) cohort (n=227) were utilized for this analysis. DNA methylation age (estimated by the Horvath method) incorporates DNA methylation status at 353 CpG sites. DNAm AA is the residual of DNA methylation age regressed on chronological age in a linear model. Spearman’s correlations and linear regression examine the relationship between DNAm AA and depressive symptoms (Beck Depression Inventory) and cardiometabolic status. The potential association and impact of SES, trauma, substance use, and stress were also considered. RESULTS/ANTICIPATED RESULTS: Contrary to our hypothesis, DNAm AA did not associate with the severity of depressive symptoms. Correlation between DNAm AA and affective symptom subscore (BDI) approached significance (p = 0.06). We observed significant correlations between DNAm AA and specific depressive symptoms including participants’ reported disappointment, disgust, or hatred toward themselves (p < 0.05), difficulty with making decisions (p < 0.05), and worry about their physical health (p < 0.05). DNAm AA was also significantly correlated with BMI (p > 0.001). Significant relationships were not evident in the subsequent regression analysis examining potential relationships between DNAm AA and depression. To our knowledge, this is the first study to examine associations between DNAm AA and depressive symptoms in AAW. DISCUSSION/SIGNIFICANCE OF FINDINGS: Depression limits life quality and quantity and is highly comorbid in CMCs. AAW have a high risk of comorbidity. This study deepens knowledge with respect to the associations between depression, CMCs, and aging with a clinically accessible marker in a population with disproportionate risk.

OBJECTIVES/GOALS: Childhood psychiatric symptoms are highly comorbid. Their co-occurrence and association with negative life outcomes is partially explained by deficits in executive control, or processes enabling self-regulation. Here, we test a novel executive neural target in three fMRI tasks and its relevance to shared psychopathology. METHODS/STUDY POPULATION: We studied 60 children [15 F/45 M; mean age (SD)=11.6 years (1.62)] with diverse diagnoses including attention deficit disorder (n=26) and autism spectrum disorder (n=22). We extracted a latent general factor of psychopathology using principal component analyses applied to parent-report Child Behavior Checklist syndrome scores. Subjects completed 3 executive control fMRI probes, tapping adaptive control, working memory, and inhibition. Correlational psychophysiological interaction (cPPI) analysis measured correlations between executive control-related modulations of activity in 414 network-affiliated parcels. We selected parcels exhibiting control-related cross-network correlations as well as control-related activity across all tasks and tested them for association with psychopathology. RESULTS/ANTICIPATED RESULTS: cPPI connectivity matrices were thresholded and graphs were identified using the Network-Based Statistic toolbox (p90th percentile PC) as well as control-related activation (>10% activated voxels; p DISCUSSION/SIGNIFICANCE: Our results examine cross-network interactions between brain regions during 3 fMRI tasks and their role in explaining individual variation in psychopathology. As executive control links to both comorbidity and life outcomes, identifying the clinically-relevant neural correlates of controlled behavior may lead to transdiagnostic treatments.

Diversification of the Translational Science workforce is a strategic goal for the National Center for the Advancement of Translational Science (NCATS) program. NCATS has identified the development of translational science education, training, and support for a diverse translational science workforce as key to advancing the growing field of translational science. An annual mixed-methods assessment has been conducted on Common Metrics data submitted by over 60 Clinical & Translational Science Awards (CTSA) programs nationwide and includes metrics addressing recruitment and retention of scientists with particular attention to underrepresented persons and women. This article describes a methodology for the development of From Insights to Action, a resource for guiding program implementation and strategic planning to develop a diverse clinical and translational science workforce. This was informed by the Common Metrics Initiative process and constituted of findings from qualitative interviews of a subset of CTSAs that participated. The dissemination of this guide had several impacts, including providing structural foci for the CTSA Fall 2020 program meeting centered on Diversity, Equity, and Inclusion in translational science; addressing NCATS’ goal of workforce diversity; and understanding the number of diverse graduates still engaged in research.

OBJECTIVES/GOALS: Precision care may engage smokers and providers in treatment but is understudied in the community. We piloted guideline-based care (GBC) alone or with Respiragene, a lung cancer polygenic risk score (PRS, 1-10), or metabolism-informed choice of medication using the nicotine metabolite ratio (NMR). METHODS/STUDY POPULATION: Daily smokers (n = 58) with stored biospecimens in the Southern Community Cohort Study were randomized 1:1:1 to GBC, PRS, or NMR, counseled to quit smoking, and co-selected FDA-approved cessation medication (nicotine replacement, varenicline) with a tobacco counselor. In PRS, precision motivational counseling was guided by PRS (i.e., lung cancer risk 10-40-fold that of never-smokers). In NMR, precision medication recommendations consisted of varenicline for faster metabolizers (NMR≥0.31) and nicotine replacement for slow metabolizers (NMR<0.31). Feasibility was defined as achieving at least 50% provider engagement (med prescription) and at least 50% patient engagement (self-reported med use). RESULTS/ANTICIPATED RESULTS: Participants were median age 59, 72% female, 81% Black, 60% with incomes <$15,000; median cigarettes/day was 15 (IQR 8-20) and 52% reported time-to-first cigarette <5 minutes, illustrating moderate nicotine dependence. Providers confirmed medication prescriptions for 40% of patients (32% GBC, 50% PRS, 37% NMR) and 83% of patients reported using medication (prescribed or unprescribed) during the study (90% GBC, 80% PRS, 79% NMR). At 6-month follow-up, 27% (n = 15) reported cessation (39% GBC, 16% PRS, 26% NMR). Among persistent smokers, 46% reported smoking at least 50% fewer cigarettes/day compared to baseline (45% GBC, 38% PRS, 57% NMR). Small sample size precluded statistical comparisons. DISCUSSION/SIGNIFICANCE OF IMPACT: Precision interventions to quit smoking are feasible for community smokers, who engaged at high rates. However, only 40% of providers supported patients’ quit attempts with medication prescriptions. Future research should test strategies to raise provider engagement in precision smoking treatment. CONFLICT OF INTEREST DESCRIPTION: R.F.T. has consulted for Quinn Emmanual and Apotex on unrelated topics. H.A.T. reported providing input on design for a phase 3 trial of cytisine proposed by Achieve Life Sciences and being a principal investigator of National Institutes of Health–sponsored studies for smoking cessation that include medications donated by the manufacturers. Other authors declare no potential conflicts of interest.

OBJECTIVES/GOALS: The first aim was to construct a controlled and high resolution FUS water tank characterization system with 1 micron step-sizes. The second aim was to create two unique standardized protocols for mapping the generated acoustic field from FUS transducers; protocol one maps the full 3D field while protocol two rapidly detects changes to the original plot. METHODS/STUDY POPULATION: To accomplish aim one, the focused ultrasound mapping platform was constructed with a water conditioning unit for water degassing and temperature control, a three-axis stage with 1 micron step-size capabilities, and a data plotting software. To measure the outcomes of aim one, the water temperature was monitored, and axis step sizes were measured through ten independent axis translation recordings. To accomplish aim two, FUS acquisitions were executed at different resolutions. For FUS localization at the cellular level, a 1-5 micron step size is required. Once the initial scan was performed, duplicate scans were executed to detect inherent perturbations or errors in the system. Once calculated, the best methods of detecting true changes to FUS signals are proposed. RESULTS/ANTICIPATED RESULTS: The FUS characterization system maintained water temperature and performed 1 micron step-sizes. While pre-existing platforms have demonstrated a resolution of one thousand recordings per cubic millimeter, the proposed system (time and computing power willing) can record one billion recordings per cubic millimeter. In practice, a resolution of 20 micron was sufficient for non-cellular level FUS characterizations. Successive 2D scans were reliably stacked to form a 3D rendering of the generated acoustic field with the average focal point intensity yielding a 1% coefficient of variation between identical scans. This inherent variation can be used as the threshold of significance for true change detection; to rapidly detect changes to the FUS signal, sampling can be performed at regions of high baseline values. DISCUSSION/SIGNIFICANCE: Focused ultrasound medical devices are gaining popularity for treatments including tumor ablation, neuromodulation, and drug delivery; however, the field lacks a standardized method to characterize these FUS transducers. The presented platform and protocols enable a rigorous and high quality translation through verification and validation.

OBJECTIVES/GOALS: The innate immune responses to Multisystem Inflammatory Syndrome in Children (MIS-C) are not fully known. Using samples from MIS-C, we will assess the cellular responses and develop a novel Tri-Specific Killer Engager (TRiKE) that engages innate immune cells to improve those responses. METHODS/STUDY POPULATION: We collected blood samples from 60 pediatric patients from which we isolated plasma and peripheral blood mononuclear cells. We received blood samples from 13 MIS-C, 32 severe acute COVID, 5 COVID-19 asymptomatic, and 15 COVID-19 negative patients. Using plasma, we then performed ELISAs to determine IgG antibody levels against SARS-CoV-2 and plaque reduction neutralization tests to determine neutralizing antibody functions. We isolated DNA to look at Fc receptor genetics. We also utilized utilize flow cytometry assays determine the phagocytosis and killing abilities of the innate cells from these patients. This data will be correlated with clinical outcomes. Additionally, we have developed a novel SARS-CoV-2 TRiKE which directs natural killer (NK) cell killing specifically to of COVID-19 infected cells. RESULTS/ANTICIPATED RESULTS: MIS-C patients had higher IgG antibody titers against SARS-CoV-2 compared to children with symptomatic or asymptomatic COVID. MIS-C patients also neutralized SARS-CoV-2 more effectively than children with acute symptomatic or asymptomatic COVID-19. We found natural killer cells and monocytes are dysfunctional in MIS-C patients and do not kill SARS-CoV-2 infected cells as well. Specifically, NK cells do not kill COVID-19 infected cells as well. To combat this, we have successfully generated and are now testing a Tri-Specific Killer engager (TRiKE) which binds one ends to NK cells, one end to the Spike protein on COVID-19 infected cells and contains IL-15 to improve NK cell function. We anticipate that we can improve NK cell killing of COVID-19 infected cells with this TRiKE. DISCUSSION/SIGNIFICANCE: We found that MIS-C patients have antibodies that can neutralize SARS-CoV-2 but that that innate immune cells that engage antibodies are dysfunctional. We are have successfully developed and are targeting this response with a TRiKE to improve innate immune cell functional; this may serve as an adjunctive therapeutic if proven successful.

OBJECTIVES/GOALS: One in six children in the U.S. has a Neurodevelopmental Disability (NDD). Prechtl’s General Movement Assessment (GMA) is a qualitative predictor of early motor dysfunction. However, no quantitative biomechanical assessment exists to more accurately identify all patients with NDD. METHODS/STUDY POPULATION: With UAMS IRB approval, as part of a larger study, healthy infants were filmed while lying supine on a force plate for 2 minutes. We studied 12 healthy full-term infants (gestational age: 38.9±1.5 weeks, age: 2.1-7.0 months; 7M, 5F; length: 64.0±5.2 cm; weight: 7.2±1.3 kg). Within our data set there were 3 infants transitioning to fidgety period (≤3 months), 4 in the fidgety period, (3-5 months), and 5 that matured beyond fidgety period (>5 months). Center of pressure (COP) path-lengths were gathered from the force plate at 1000 Hz. We grouped our data with K-means clustering and performed statistical analysis with ANOVA. RESULTS/ANTICIPATED RESULTS: We divided our data into 3 distinct clusters. The first group contained infants with moderate variability of movements which included 2 infants between 3 and 5 months and 2 infants slightly outside of this range. The second group, with mild variability in movements, included 4 infants between 2 and 3 months as well as 2 infants just older than 5 months. The third group, with little variability in movements, included 2 infants older than 5 months. A GMA reader (TJ) qualitatively confirmed these findings with video footage. Using a threshold of p<0.05, data sets within the clusters were similar and significantly different from other clusters. DISCUSSION/SIGNIFICANCE OF IMPACT: Fidgety infants have greater variability in COP patterns than their mature counterparts. We anticipate additional COP measurements will correspond with qualitative GMA analyses. Artificial Intelligence-based quantification of the GMA may be useful in earlier detection or prediction of NDD outcomes.

OBJECTIVES/SPECIFIC AIMS: This study aims to understand the utility of 5-ALA and SWIG in detecting areas of neoplasm in a murine model of GBM. Primary outcome is the distribution of the two dyes in comparison to the true tumor extent; the sensitivity, specificity, PPV, and NPV of both dyes will be calculated. The secondary outcome is the suitability of existing cell-lines used for GBM research for studies in fluorescence-guided surgery. METHODS/STUDY POPULATION: Two cell lines are used for this research: U87, derived from human GBM; and GL261, derived from rodent stem cells. U87 are implanted intracranially into 6-week old athymic, nude, female mice, while GL261 are implanted intracranially into 10-week old female C57BL/6 mice. The mice are weighed every 3 days to monitor health and bioluminescence imaging is performed between 7-10 days after implantation to confirm tumor implantation and monitor tumor growth. The mice are sacrificed between 10-21 days after implantation. 5mg/kg of intravenous ICG is administered 24-hours prior to harvest and 250mg/kg of intraperitoneal 5-ALA is administered 3-hours prior to harvest. Once the mice are sacrificed, their brains are quickly harvested and placed in cold formalin. Using a high-resolution Odyssey CLx scanner, near-infrared fluorescence from ICG is captured in coronal cross sections of the brains through the tumor. Similarly, 5-ALA fluorescence is imaged using a 405nm LED excitation source and 610-690nm bandpass filter. Afterwards, slices of the brain are stained with H&E, which serves as the gold-standard of the extent of tumor. Images from ICG, 5-ALA, and H&E can then be compared using ImageJ to compare the extent of tumor to the distribution of the dyes. RESULTS/ANTICIPATED RESULTS: In separate, previous studies in humans, both 5-ALA and SWIG have demonstrated utility in detecting residual neoplasm in HGG resections. In general, 5-ALA is more specific for areas of neoplasm, while SWIG is more sensitive. Thus, I anticipate that in this study, SWIG will show a greater distribution than 5-ALA, with SWIG distributing to areas beyond the tumor and 5-ALA distributing within, but not completely covering, the tumor. SWIG’s sensitivity and NPV for detecting tumor should be >90%, while its specificity and PPV may be closer to 50%. For 5-ALA, specificity and PPV should be close to 80-90%, but its sensitivity and NPV may be <50%. In terms of cell-line, preliminary results suggest that U87 cells are not suitable for research involving 5-ALA. We suspect that this is partly due to the limited infiltrative nature of U87 cells; in fact, the cells form a spherical mass, imitating metastases rather than true HGGs. The U87 masses do not have significant vascularity, which likely limits the amount of 5-ALA that can distribute to inside the tumor. DISCUSSION/SIGNIFICANCE OF IMPACT: 5-ALA is currently the only FDA-approved agent for fluorescence-guided neurosurgery. However, it has multiple limitations, which ultimately results in its low sensitivity and NPV. Our novel technique, which has demonstrated much higher sensitivity at the cost of specificity, offers an alternative that may help surgeons better achieve total resections in the operating room. These two agents have not been compared directly in humans or mice. Thus, this experiment sets up an important precedent, on which a human clinical trial comparing the two agents’ effects on resection rates and patient outcome can be performed. Ultimately, this work will lay the foundation for future research into fluorescence-guided neurosurgery, both in the visible and NIR spectrum.

ABSTRACT IMPACT: The pathophysiologic features of a metabolomic endotype that predicts patient outcomes due to sepsis have the potential to direct new therapies that target immune dysregulation and bioenergetic insufficiency. OBJECTIVES/GOALS: Acute respiratory failure (ARF) requiring mechanical ventilation is a frequent complication of sepsis and other disorders. It is associated with high morbidity and mortality. Despite its severity and prevalence, little is known about metabolic and bioenergetic changes that accompanying ARF. METHODS/STUDY POPULATION: In this study, semiquantitative and quantitative ultrahigh performance liquid chromatography mass spectrometry (UHPLC MS) analysis was performed on patient serum collected from the Trial with Acute Respiratory failure patients: evaluation of Global Exercise Therapies (TARGET). Serum from survivors (n=15) and nonsurvivors (n=15) was collected at day 1 and day 3 after admission to the medical intensive care unit as well as at discharge in survivors. Pathway analysis of the biochemical changes was performed to determine whether the disruption in specific metabolic pathways can identify the bioenergetic and metabolomic profile of these patients. RESULTS/ANTICIPATED RESULTS: Significant metabolomic differences were related to biosynthetic intermediates of redox cofactors nicotinamide adenine dinucleotide (NAD) and NAD phosphate (NADP), increased acyl-carnitines, and decreased acyl-glycerophosphocholines in nonsurvivors compared to survivors. The metabolites associated with poor outcomes are substrates of enzymatic processes dependent on NAD(P), while the abundance of NAD cofactors rely on the bioavailability of dietary vitamins B1, B2 and B6. Changes in the efficiency of the nicotinamide-derived cofactors’ biosynthetic pathways also associate with an alteration of the glutathione-dependent drug metabolism as characterized by the substantial differences observed in the acetaminophen metabolome. DISCUSSION/SIGNIFICANCE OF FINDINGS: This metabolomic endotype represents a previously unappreciated association between severity of outcomes and micronutrient deficiency, thus pointing to new pharmacologic targets and highlighting the need for nutritional remediation upon hospitalization to improve patient outcomes due to ARF.

OBJECTIVES/GOALS: The goal of this project was to perform an exploratory analysis of the research themes of scientific publications from the Miami Clinical and Translational Science Institute using text mining techniques and using bibliometric characterization and network analysis to further assess research trends. METHODS/STUDY POPULATION: Publications were identified from the Web of Science database using Miami CTSI grant numbers as search criteria for the period 2013-2021 and KL2 scholar publications. Following data pre-processing, topic modeling was performed using the Latent Dirichlet Allocation algorithm and cluster analysis in the R programming language. The resulting themes will be further analyzed by employing a citation-based impact assessment approach to identify trends over time. Network analysis of publications will be performed using the VOSviewer package to visualize publication networks using citation and co-authorship relations within each major theme and their evolution over time. Findings will be evaluated for alignment with institutional research strategy. RESULTS/ANTICIPATED RESULTS: About 400 CTSI publications from 2013-2021 to date were used for analysis. Twenty topics and five major research themes were identified among the Miami CTSI publications – neuroscience, cancer, community and public health, metabolics, and HIV/infectious diseases. Top keywords within each topic were aligned with the most frequent author-assigned keywords for that topic. The CTSI research themes were also well-aligned with the institutional vision for research and focus areas. Trends using citations and networks for each research theme are currently being analyzed and results will be included in the overall findings post analysis. DISCUSSION/SIGNIFICANCE: Text mining was successfully used in identifying topics and research themes for clinical and translational research publications of the Miami CTSI, and in combination with bibliometric characterization, will be helpful in shaping CTSI strategy and alignment with the universitys research priorities.

OBJECTIVES/GOALS: Age-related lobular involution (LI) is a physiological process of breast epithelial regression that occurs primarily during perimenopause (ages 45-55); women in this age range for which the process of LI is delayed, defined as LI stagnation, show significantly increased risk of breast cancer as compared to LI progression patients. METHODS/STUDY POPULATION: The Mayo Clinic Benign Breast Disease (BBD) cohort includes ~1000 women who had multiple sequential benign biopsies. 103 patients were found to have sequential biopsies during the perimenopausal period, of which 10 eventually progressed to breast cancer. These patients were assessed for LI stagnation vs LI progression by quantifying 10 lobules per slide and comparing median acini number and median lobule size between initial and subsequent biopsies from the same patients. RNA was derived from whole tissue sections from the initial biopsies, and profiled using NanoString IO360 and BC360, which were normalized using RUVg methods. Differentially expressed genes associated with LI stagnation were defined as having two-tailed, unpaired p-values less than 0.05. RESULTS/ANTICIPATED RESULTS: Analysis showed subsetting patient sets by time between biopsies improves classification of stagnant vs. progression. Differential gene analysis identified 37 genes associated with LI stagnation and LI progression, and 20 of these genes were found to overlap a set of 128 gnese that were differentially expressed between women who subsequently developed breast cancer vs remained cancer-free. These genes represent potential biomarkers of processes that link LI stagnation and increased breast cancer risk. DISCUSSION/SIGNIFICANCE: In future studies, we intend to study these genes that were shown to be upregulated in LI stagnation for their association with subsequent development of breast cancer in independent cohorts of women with BBD. We will use this knowledge to improve individualized risk assessment, which will help focus surveillance and prevention strategies.

OBJECTIVES/GOALS: Weaimed to compare subjective and objective sleep in individuals with chronic post-concussive symptoms. We hypothesized an association between self-reported sleep quality and objective sleep parameters, which is different for concussed and control cohorts. METHODS/STUDY POPULATION: 28 individuals with chronic post-concussive symptoms and 13 age-matched controls (no concussion history) completed the ISI, PSQI, PROMIS Depression, Anxiety, Stress and Cognitive questionnaires at enrollment. Objective sleep parameters were obtained for a minimum of 7 days and up to 30 days with a validated sleep monitoring device placed under the subject’s bed (Emfit). For each night, raw activity data per minute were analyzed to determinein-bed, sleep, wake, andout-of-bedtimes. These measures were used to calculate total sleep time (TST), sleep onset latency (SOL), and wake after sleep onset (WASO) for each night. RESULTS/ANTICIPATED RESULTS: Concussed individuals reported worse sleep with PSQI and ISI scores significantly higher than controls. They also showedsignificant associations between PSQI and Dep ression, ISI and Depression, and ISI and Anxiety scores. There was no difference between objective sleep parameters in the concussed and control cohorts (in-bed/sleep/wake/out-of-bed times, TST, SOL, and WASO). Instead, higher PSQI, ISI, Depression, Anxiety, and Stress scores (greater symptom burden) were all associated with later sleep times, where as higher Cognitive scores (greater cognitive function) were associated with earlier sleep times, regardless of group status. DISCUSSION/SIGNIFICANCE: Concussed individuals report worse subjective sleep but no differences to controls when objectively assessing sleep. Depression/anxiety, and not concussion status, determine objective sleep parameters. Psychiatric comorbidities should inform the treatment of post-concussive sleep disturbances.