OBJECTIVES/GOALS: Primary graft dysfunction (PGD) is acute lung injury in the first three days after lung transplant. Patients that experience PGD have increased mortality and an increased risk of chronic lung allograft dysfunction. The pathogenesis is thought to be an ischemia-reperfusion injury but is incompletely understood and there are no specific therapies. We investigated the role of the microbiome in PGD and associations with inflammation and markers of aspiration. METHODS/STUDY POPULATION: We collected airway lavage samples from lung transplant donors before procurement and recipients after reperfusion. We extracted DNA, amplified the bacterial 16S rRNA gene, and sequenced on the Illumina MiSeq platform. QIIME2 and Deblur were used for bioinformatic analysis. R packages were used for downstream analysis and visualizations. The host response was quantified using the Milipore 41-plex Luminex and an ELISA for pepsin. Clinical data was collected by the Penn Lung Transplant Outcomes Group. PGD was assessed by degree of hypoxemia and chest X-ray findings in the 72 hours after transplant. RESULTS/ANTICIPATED RESULTS: There was no significant difference in alpha diversity (Shannon index, p = 0.51), biomass (via comparison of 16S amplicon PicoGreen, p = 0.6), or beta diversity (Weighted UniFrac, p = 0.472, PERMANOVA) between subjects with PGD grade 3 (n = 36) and those that did not (n = 96). On taxonomic analysis, we found an enrichment of Prevotella in donor and recipient lungs that went on to develop PGD (p = 0.05). To follow up this finding we measured immune response and pepsin concentrations in recipient lungs. We found elevated levels in 35/41 cytokines measured in subjects that developed PGD as well as an elevation in pepsin and a correlation between pepsin concentration and Prevotella relative abundance (Figure 1). Additionally, Prevotella relative abundance had statistically significant positive correlations with multiple cytokines such as IL-6 (Pearson’s = 0.26, p = 0.009) and eotaxin (Pearson’s = 0.24, p = 0.016). DISCUSSION/SIGNIFICANCE OF IMPACT: There is an enrichment of oral anerobes in lung allografts that eventually develop PGD. This is associated with elevated levels of pepsin and markers of inflammation. These lines of evidence suggest aspiration contributes to priming the allograft for PGD.

OBJECTIVES/SPECIFIC AIMS: N/A. METHODS/STUDY POPULATION: N/A. RESULTS/ANTICIPATED RESULTS: N/A. DISCUSSION/SIGNIFICANCE OF IMPACT: There is an increasing need to foster cross-disciplinary research to address complex problems within healthcare. The Sinai Team-based Translational Education Program: the URM Propeller (STTEP-UP) is a NCATS funded program through the Icahn School of Medicine at Mount Sinai. Its goal is to facilitate URM post-doctoral trainees becoming innovative leaders in clinical and translational research. The program includes a team-based research component, where fellows collaborate on a project. This year, disciplines represented by the four fellows include Cardiology, Psychiatry, Neurology, and Pediatrics. Identifying a clinical question and designing an investigation was facilitated by group brainstorming meetings with program mentors. Fellows designed a project to identify medical testing and prescribing that were not clinically indicated throughout the healthcare system, with the goal of exploring whether an intervention, including provider education, could reduce ordering practices. In addition to regular in-person meetings, a licensed virtual learning environment and free web-based sharing platform were used to foster collaboration. Challenges faced throughout this process, included fellows struggling to find protected time, difficulties accessing broad sets of data across the healthcare system, and overcoming administrative barriers between departments. Strengths of this approach, included fellows learning new research strategies and feeling a deeper sense of commonality with their peers. Overall, this experience supports the idea that cross-disciplinary research improves the collaboration and education of emerging researchers. However, addressing logistical and systems-based barriers may better facilitate this education and research.

OBJECTIVES/SPECIFIC AIMS: (1) Obtain publically available citation data, funding data, and generate multiple networks topologies based on dynamic queries of individual faculty. (2) Determine successful pathways that lead to tenure, and career advancement, in addition to determining the effect of CTSA programs on faculty collaboration. (3) Develop publically available commercial interface for the study of faculty networks METHODS/STUDY POPULATION: For our study we included all available citation and funding data publically available on all CTSA programs (as of 2015) with historical data dating back to 2005. We then included the top 25 collegiate institutions who may not have had a CTSA program (eg, Princeton University). We then developed network topologies for each university network, and explore the evolution of individuals in these networks, and the effects of faculty development—as an example in the University of Rochester network, we singled out the directors of the CTSA program there to understand their level of centrality and overall impact on network development, with key observations being that early publications across varying domains lead to stronger network performance. Although individuals who did not benefit from such development, may have succeeded but if they did were likely to leave the institution for elsewhere. RESULTS/ANTICIPATED RESULTS: A secondary goal of this project is to evaluate the effectiveness of the Clinical & Translational Science Institute (CTSI) since its inception in 2006. The mission of CTSI is to advance the field of translational science and research, to link other departments at URMC and community stakeholders by research collaboration, publication, and goals to improve population health, and provide translational education and training to students, researchers, and physicians. To determine how the induction of CTSI affects collaboration within the URMC network, we examined the role of funding in the CTSI network. This was done around the second successful funding around 2013. In doing so we can see that not only did the funding request affect the network topology, but opened new collaborations which were not present prior to the request. DISCUSSION/SIGNIFICANCE OF IMPACT: We have developed an automated method, which is superior to manual methods necessary for citation generation and funding data analysis of faculty growth in citation networks. This technique is applicable to all institutions, not just those in a CTSA environment, but demonstrates the benefit of cross-collaborative efforts, in the case of the URMC network we can state the following. The key takeaway is for individuals to succeed in the URMC collaborative environment they should create their own network and expand it and eventually rise to prominence. There are 2 pathways to this you can take the Dewhurst approach which is to seek out collaborations among internal peers and scale up. Or you can take the Nedergaard approach which is develop the special network, and gain enough public recognition outside of the network that you are capable of leaving it (Fig. 2d). In either case, collaborations among communities and diverse out-degree networks allow faculty to succeed in their given field. Given the wealth of data which has been curated in this fashion, there are numerous explicit questions that can be asked of the data. One of the unique approaches of this data is that is highly reproducible, which allows various questions to be asked. Future work would try to determine what optimal pathways are in a given network to success, and who are ideal collaborators, and collaborations to avoid. Given this information, custom pathways to career success for individual faculty can be developed, moving beyond purely institutional level co-citation networks, which do little to advance faculty development at scale. In Figs 1c and d, the network increased by 75% in terms of graph density (0.007) and decreased by 18.8% (16) in terms of diameter. What this suggest in that the interconnectivity of the network grew dramatically, while the ability for new members to integrate into it increased. This also apparent when one examines the modularity of the network down by 3.6% (0.857), this suggest that the network has as many communities but these communities are less isolated that those in the previous funding year, meaning fields are becoming more transdisciplinary in their collaborations. This was the result of the presence of a CTSA program, thus demonstrating the effectiveness of such institutions, however, our analysis also lays the framework for applying this to other institutions which may be considering a CTSA. Or maintaining the success of a given CTSA program, and ultimately determining where faculty should place their efforts and choose which programs to pursue career advancement.

OBJECTIVES/GOALS: Clinical translational studies inform clinical practice patterns through dissemination of clinical practice guidelines (CPG). In EM practices change to rapidly for timely local EHR implementation. We test the OMG BPM+Health specification for rapid deployment of best practices relevant to EM. METHODS/STUDY POPULATION: The OMG Business Process Management for Healthcare (BPM+Health) specification combines BPMN™ with Case Management Model and Notation (CMMN™) and Decision Model and Notations (DMN™) to “disseminate and leverage evidence-based best-practices at the point of care.” The American College of Emergency Physicians (ACEP) Board-certified Emergency Physicians modeled practice guidelines in the BPM+ modeling language during on-line meetings. Two common emergency conditions were selected for initial pilot testing: 1) evaluation and treatment of first trimester bleeding in pregnant patients, and 2) the evaluation and treatment of non-traumatic low back pain. RESULTS/ANTICIPATED RESULTS: The protocols were successfully modeled during four on-line meetings in less than 2 months. Process steps from initial evaluation to disposition were implemented using BPMN™. When clinicians need to evaluate the patient to collect data for decision making the inputs and outputs were modeled in CMMN™. Decision logic is represented as DMN™. The software tool linked the components for easy browsing and authoring the logic. The Physicians easily followed the displayed logic. The practice recommendations from each policy were successfully modeled, using the standard BPM+ notation to support rapid implementation in EHRs. Detailed implementation specifications will be shared. DISCUSSION/SIGNIFICANCE OF IMPACT: This pilot project demonstrated the feasibility of the OMG approach to solving Clinical Practice Guideline Implementation and Dissemination Barriers. Ongoing work by involved specialty societies will be necessary to demonstrate the scalability and sustainability of this approach.

OBJECTIVES/GOALS: Mobile health applications are widely used by the public but vary in how they are classified and regulated. This study examines the evidence of the safety and efficacy of mobile medical applications specifically focusing on those that are used to manage diabetes. METHODS/STUDY POPULATION: To understand the current regulatory landscape of mobile health applications (mHealth apps) for diabetes, a literature survey was conducted using the Pubmed database. Top mHealth apps were identified by searching the Apple store website using 10 key terms associated with diabetes management applications. A maximum of ten results, when available for each key term, were studied by exploring the FDA databases to understand how the products were regulated and if any were subject to recalls. These selected mHealth apps were also searched on clinicaltrials.gov to see if there were ongoing or completed clinical trials and if the trials were designed to include efficacy and safety outcome measures. RESULTS/ANTICIPATED RESULTS: Of the 71 mHealth apps for diabetes management that were identified, 16 were regulated. These products spanned a diverse range of functions including device data and decision support systems. Although 11 had clinical trial data demonstrating efficacy, only 4 had data demonstrating both efficacy and safety. Two of the regulated applications were subject to product recalls due to programming errors that resulted in incorrect insulin dose recommendations. These two applications had clinical trials evaluating efficacy but not safety. The companies noted that the incorrect insulin calculation from their respective mHealth app could cause either a low- or high-impact hypoglycemic event. DISCUSSION/SIGNIFICANCE OF IMPACT: With little to no clinical trial data to support their safety and efficacy, mHealth apps in the diabetes marketplace pose risks for patients as evidenced by recent safety-related recalls. The results of this study indicate that these products may need to be more tightly regulated.

ABSTRACT IMPACT: Impaired neuromuscular control could lead to the failure in activation and deactivation of the target muscles in a timely manner, with the concurrence of activities of non-targeted muscles. OBJECTIVES/GOALS: Stroke leads to impaired capacity to manipulate objects with the hand in terms of timing and strength of grasp. The influence of abnormal coupling across more proximal arm muscles post stroke on the failure in normal functioning of finger flexor muscle activity is of interest to investigate. METHODS/STUDY POPULATION: We have recruited 12 participants with stroke hemiparesis in the sub-acute or chronic stage. Motor impairment of the arm was assessed using electromyography (EMG) and the Fugl-Meyer Upper Extremity (FMUE) assessment. Participants were requested to flex and relax the metacarpophalangeal (MCP) joints against motorized resistance in response to audible tones to determine timing and strength during flexion. They were asked to flex maximally, as quickly as possible, in response to the first of a pair of tones, and relax as quickly as possible after the second tone. Delays in initiation and termination were evaluated using EMG responses versus a predefined threshold. RESULTS/ANTICIPATED RESULTS: We anticipate greater delays in grasp initiation as well as in grasp termination in participants with a greater extent of abnormal coupling across more proximal muscles of the upper extremity in comparison to participants with a less extent, using the results of the FMUE assessment. Also it is expected that participants with a greater extent of the flexion synergy produce a less extent of force generation. The EMG results will show that activities of more proximal arm muscles precede the initiation of MCP flexion and their activity termination precedes that of MCP flexion, significantly more in participants with a greater extent of the flexion synergy. DISCUSSION/SIGNIFICANCE OF FINDINGS: The flexion synergy over the arm following stroke affects the timing and strength of hand grasp. Impaired neuromuscular control could lead to the failure in activation and deactivation of the target muscles in a timely manner, with the concurrence of activities of non-targeted muscles.

OBJECTIVES/GOALS: This work aims to explore how citizen science serves as a transformative frame work to bridge scientific knowledge, focusing on its potential to enhance transdisciplinary learning in artificial intelligence (AI) biomedical and clinical sciences by facilitating near-peer mentoring. METHODS/STUDY POPULATION: Our group of eight friends comprise a multicultural and multidisciplinary cohort including students from the USA, Philippines, Indonesia, and Guatemala pursuing PhD degrees in electrical and computer engineering, epidemiology, physics, and MD, PharmD, and DMD degrees. We engage in shared online courses, collaborative projects, and abstract submissions. Employing our collective knowledge, we design interactive learning experiences, support each other’s initiatives, and collaboratively develop lectures and presentations. We in tend to expand collaborations in biomedical AI education while fostering principles of experiential and collaborativelearning, constructivism, and authentic inquiry. RESULTS/ANTICIPATED RESULTS: Our recent successes include submittedconference abstracts on data science and AI education in pharmacy and the facilitation of a guest lecture in health informatics. Additionally, we are currently collaborating on seven biomedical machine learning projects in radio frequency engineering, aiming for conference submissions. Moving forward, our goal is to expand our group, support the formation of similar communities, and promote data science and AI literacy in biomedical and clinical contexts. We aspire to extend this knowledge to families, classmates, and eventually patients, facilitating a broader understanding of the role of AI in healthcare. DISCUSSION/SIGNIFICANCE: We believe diverse expertise and pedagogical theories can help demonstrate the potential of citizen science to democratize scientific experience. By nurturing collaborative networks our efforts aim to bridge gaps between disciplines and enhance the broader public’s understanding of AI in healthcare.

Availability of trained professionals to assist researchers navigating regulatory pathways for new drug and device development is limited within academic institutions. We created ReGARDD (Regulatory Guidance for Academic Research of Drugs and Devices), a regional forum initially involving regulatory professionals from four Clinical and Translational Science Award (CTSA)-funded institutions, to build and capitalize on local expertise and to develop a regulatory guidance website geared toward academic researchers. Since 2015, members organized 15 forums covering topics such as FDA premarket submissions, gene therapy, and intellectual property for devices and therapeutics. Through user feedback, targeted surveys, and ongoing iterative processes, we refined and maintained a shared regulatory website, which reached 6000+ users in 2019. Website updates improved navigation to drug versus device topic areas, provided new educational content and videos to address commonly asked questions, and created a portal for posting upcoming training opportunities. Survey respondents rated the website favorably and endorsed expanding ReGARDD as a centralized resource. ReGARDD strengthened the regional regulatory workforce, increased regulatory efficiency, and promulgated best organizational and operational practices. Broad-scale deployment of the ReGARDD model across the CTSA consortium may facilitate the creation of a network of regional forums and reduce gaps in access to regulatory support.

Adenylate kinases (AKs) are important enzymes involved in cellular energy metabolism. Among AKs, AK5 (adenylate kinase 5), a cytosolic protein, is emerging as a significant contributor to various diseases and cellular processes. This comprehensive review integrates findings from various research groups on AK5 since its discovery, shedding light on its multifaceted roles in nucleotide metabolism, energy regulation, and cellular differentiation. We investigate its implications in a spectrum of diseases, including autoimmune encephalitis, epilepsy, neurodegenerative disorders such as Alzheimer’s and Parkinson’s, diabetes, lower extremity arterial disease, celiac disease, and various cancers. Notably, AK5’s expression levels and methylation status have been associated with cancer progression and patient outcomes, indicating its potential as a prognostic indicator. Furthermore, AK5 is implicated in regulating cellular processes in breast cancer, gastric cancer, colorectal carcinoma, prostate cancer, and colon adenocarcinoma, suggesting its relevance across different cancer types. However, a limitation lies in the need for more robust clinical validation and a deeper understanding of AK5’s precise mechanisms in disease pathogenesis, despite its association with various pathophysiological conditions. Nonetheless, AK5 holds promise as a therapeutic target, with emerging evidence suggesting its potential in therapy development.

ABSTRACT IMPACT: This work examines the association between diabetes mellitus and latent tuberculosis infection among a cohort of household contacts exposed to active tuberculosis in Ethiopia, focusing attention on the need for further translational research to determine the mechanisms of susceptibility to Mycobacterium tuberculosis infection among individuals with diabetes and pre-diabetes. OBJECTIVES/GOALS: Diabetes mellitus (DM) is an established risk factor for active TB disease, but there is limited understanding of the relationship of DM and latent tuberculosis (LTBI). We sought to determine the relationship between DM or pre-DM with LTBI among household or close contacts (HHCs) of active TB cases in Ethiopia. METHODS/STUDY POPULATION: We conducted a cross-sectional study of the HHCs of index active TB cases enrolled in an ongoing TB Research Unit (TBRU) study in Addis Ababa, Ethiopia. HHCs of individuals with laboratory-confirmed TB had QuantiFERON ®-TB Gold Plus (QFT) and glycated hemoglobin (HbA1c) tests performed. LTBI was defined as a positive QFT and lack of symptoms. HbA1C results were used to define no DM (HbA1c <5.7), pre-DM (HbA1c 5.7-6.5%), and DM (HbA1c >6.5% or prior history of diabetes). Logistic regression was used to estimate adjusted odds ratios (OR) and 95% confidence intervals (CI) after adjustment for age, sex and HIV status as potential confounders. RESULTS/ANTICIPATED RESULTS: Among 466 HHCs, the median age was 29 years (IQR 23-38), 58.8% were female, 3.4% were HIV-positive, and median BMI was 20.9 kg/m^2 (IQR 18.9-23.8). Overall, 329 HHCs (70.6%) had LTBI, 26 (5.6%) had DM and 73 (15.7%) had pre-DM. Compared to HHC without DM, the prevalence of LTBI was higher in those with pre-DM (68.9% vs. 72.6%; OR 1.19, 95% CI 0.69-2.13) and those with DM (88.5%; OR 3.45, 95% CI 1.17-14.77). In multivariable analysis, there was a trend towards increased LTBI risk among HHCs with DM vs. without DM (OR 2.16, 95% CI 0.67-9.70) but the difference was not statistically significant. Among HHCs with LTBI, the median IFN-? response to TB1 antigen was modestly greater in those with DM (5.3 IU/mL; IQR 3.0-7.8) and pre-DM (5.4 IU/mL; IQR 2.0-8.4) compared to HHCs without DM (4.3 IU/mL; IQR 1.4-7.7). DISCUSSION/SIGNIFICANCE OF FINDINGS: Our results suggest that DM may increase the risk of LTBI among HHCs recently exposed to active TB. Among those with LTBI, increased IFN-? antigen response in the presence of DM and pre-DM may indicate an exaggerated but ineffectual response to TB. Further investigation is needed to assess how dysglycemia impacts susceptibility to M. tuberculosis.

OBJECTIVES/SPECIFIC AIMS: Community engagement is a commonly used term, but is complex in both meaning and application. In order to help academic institutions and administrators develop infrastructure to promote and support community engagement and to help investigators work productively with communities, this analysis discusses the major components of community engagement in research on both the institutional and individual project levels as well as the interplay between them. METHODS/STUDY POPULATION: A literature synthesis conducted by a community engagement in research committee at 1 CTSA institution that examined the myriad factors related to effective community engagement in research identified across multiple disciplines was used to distill the major factors identified, assesses the interplay of the identified factors, and produce a conceptual model to help administrators and investigators apply best practices in engaging communities in clinical and translational research. RESULTS/ANTICIPATED RESULTS: This work takes a concept—community engagement in research—that is often stated and discussed, but is highly complex and challenging to implement—and identifies and discusses the multiple, interrelated factors germane to it. The model illustrates that while community engagement in research is implemented in the context of individual projects, a deep and continual interplay between individual projects and the goals, capacity, and policies of research institutions is needed for rigorous, ethical, and effective community engagement. DISCUSSION/SIGNIFICANCE OF IMPACT: Results are presented through a conceptual framework which displays the major components needed for rigorous, ethical, and effective community engagement in clinical and translational research. In addition, the conceptual framework presented will provide assistance to those developing approaches to measure and evaluate institutional readiness for community engagement in research as well as the effectiveness of individual community engagement efforts.

OBJECTIVES/GOALS: Ocular graft versus host disease (oGVHD) affects ~50% of individuals after an allogeneic hematopoietic stem cell transplant for treating blood cancers. OGVHD results in severe dry eye disease and decreased vision. Quantifying specific cytokine changes in tears may reveal biomarkers and future treatment targets for patients with oGVHD. METHODS/STUDY POPULATION: The goal of this study is to determine if cytokines can be measured in tears with the Isoplexis platform. This pilot validation study evaluates the tears of a patient with oGVHD utilizing the Isoplexis platform. The Isoplexis has specific advantages for tear samples including high-throughput analysis, and small sample requirements, but has yet to be validated in tears. A sample from normal and oGVHD patient tears were collected for comparison. Samples were analyzed on two separate backgrounds–standard Bovine Serum Albumin (BSA) background and artificial tears (ATs). The negative control was ATs and positive control was a concentrated cytokine solution. Analysis of 22 cytokines was performed. RESULTS/ANTICIPATED RESULTS: Analysis of 22 cytokines was performed. As expected, the cytokine levels of the ATs alone were below the limit of detection (LOD). The oGVHD patient tears showed elevated TNF-alpha, TNF-beta, perforin, MIP-1a, MIP-1β, MCP-1, IL2, IL4, IL5, IL-7A, IL9, IL-13, IL-15, IFN-γ, granzyme B, and GM-CSF with ATS background, but no cytokines above the LOD in the BSA background plate. The control tears had elevated IP-10. The elevated cytokines for the oGVHD patient corresponded to symptom severity and clinical findings. DISCUSSION/SIGNIFICANCE: These results suggest that using ATs as the background with the Isoplexis platform improves the sensitivity to detect tear cytokines. Findings of elevated IL-7A and GM-CSF in tears parallels literature findings for oGVHD. Further evaluation of samples will continue to validate the Isoplexis multiplex assay for tear cytokine analyses.

OBJECTIVES/GOALS: The Clinical Implementation stage in the translational pipeline is hampered by the tension between formal evidence and clinician perceptions. For instance, when guidelines are translated into electronic clinical decision support alerts, they are often ignored. Using advances in LLMs we present a framework to quantify these discrepancies. METHODS/STUDY POPULATION: We hypothesize that ignoring guideline-based alerts may be driven by discordances between clinical guidelines’ deterministic realities and clinician’ perception of clinical reality. Until now this has been very difficult to measure using quantitative methods. We argue that advances in Large Language Models (LLM) provide an avenue for exploring this quantitatively. Here we present the method and preliminary results comparing the responses of BioBERTT from a carefully designed set of questions when the LLM is fine-tuned using either formal guidelines or transcripts of clinicians discussing guidelines and clinical care in the parallel domain. The formal “distance” between the LLM responses is evaluated using quantitative metrics like the Hamming Distance. RESULTS/ANTICIPATED RESULTS: We present a description of the architecture used to prove or disprove our hypothesis. We will present results obtained when training the architecture with data that could be used to test the limits of our hypothesis, by fine-tuning BioBERT with diverse synthetic clinical views, either in agreement or disagreement with the formal guidelines. Results comparing sepsis guideline text with transcripts of interviews with Emergency Department clinicians discussing care practices for sepsis in the ED transcripts will also be considered. Our current emphasis is on securing a wider range of transcripts of clinicians interviewed from different clinical specialties and different clinical settings. While here we focus on clinical guidelines, the framework supports any intervention in the Clinical Implementation stage. DISCUSSION/SIGNIFICANCE: Leveraging recent advances in LLMs, we develop a framework that can quantitatively measure the differences between guidelines and clinician perception of best practices. We demonstrated the functionality of this approach using synthetic data and initiated the collection of clinician transcripts to test the framework in real clinical situations.

Cardiovascular disease (CVD) is largely preventable, and the leading cause of death for men and women. Though women have increased life expectancy compared to men, there are marked sex disparities in prevalence and risk of CVD-associated mortality and dementia. Yet, the basis for these and female-male differences is not completely understood. It is increasingly recognized that heart and brain health represent a lifetime of exposures to shared risk factors (including obesity, hyperlipidemia, diabetes, and hypertension) that compromise cerebrovascular health. We describe the process and resources for establishing a new research Center for Women’s Cardiovascular and Brain Health at the University of California, Davis as a model for: (1) use of the cy pres principle for funding science to improve health; (2) transdisciplinary collaboration to leapfrog progress in a convergence science approach that acknowledges and addresses social determinants of health; and (3) training the next generation of diverse researchers. This may serve as a blueprint for future Centers in academic health institutions, as the cy pres mechanism for funding research is a unique mechanism to leverage residual legal settlement funds to catalyze the pace of scientific discovery, maximize innovation, and promote health equity in addressing society’s most vexing health problems.

OBJECTIVES/SPECIFIC AIMS: A hallmark of progressive HIV-1 infection is the massive activation and depletion of the gut barrier protective CD4 T helper subsets (Th17 and Th22) in the intestinal mucosa. The loss of these cells is thought to contribute to microbial translocation and systemic immune activation that occurs during chronic infection. In addition to the loss of protective Th subsets, we previously showed that chronically HIV-1 infected individuals have an altered colonic mucosal microbiome, which is in part characterized by a lower relative abundance of bacteria that produce the short-chain fatty acid butyrate in conjunction with increased relative abundance of gram-negative pathobionts. This dysbiosis was linked to markers of mucosal and systemic immune activation in these individuals. Following up on these clinical observations, we sought to understand how a loss of butyrate might contribute to HIV-associated inflammation. Initial studies showed that the addition of butyrate to cultured lamina propria mononuclear cells (LPMC) resulted in decreased pathobiont-driven gut T cell activation, HIV-1 infection levels and production of IL-17 and IFNy. Since the gut barrier protective Th17 and Th22 subsets are preferentially infected and depleted, which is critical to HIV-1 pathogenesis, we wanted to determine the mechanism by which butyrate modulates activation of these important Th subsets in the gut. METHODS/STUDY POPULATION: Total LPMCs or purified LP CD4 T cells were isolated from human jejunal tissue (n=3–6), labeled with CFSE and cultured with TCR/CD28 beads to mimic APC driven T cell activation, with the addition of butyrate at physiologic doses(0–2 mM). Four days after culture, secreted cytokine(IL-17 and IFNy) levels were measured by ELISA. Cells were then short-term (4 hr) mitogenically stimulated (PMA/Ionomycin) in the presence of a golgi transport inhibitor. Total CD4 T cell activation (CD38+/HLA-DR+, CD25+), proliferation (CFSElow), and frequencies of intracellular cytokines were measured by multi-color flow cytometry. Paired t-tests were performed to determine statistical significance. RESULTS/ANTICIPATED RESULTS: Butyrate inhibited LP CD4 T cell activation (p=0.013) and proliferation (p=0.015) within total LPMCs stimulated with TCR/CD28 beads in a dose-dependent manner, with significant activity starting at 0.125 mM. Quantification of total secreted cytokines revealed that butyrate significantly decreased both IL-17 and IFNy production after 4 days of culture at 0.0625 mM and 0.25 mM of butyrate, respectively. Assays using purified LP CD4 T cells demonstrated that butyrate directly decreased LP CD4 T cell activation, proliferation and cytokine production in response to TCR/CD28 stimulation. Studies on specific T helper subsets revealed that butyrate inhibited proliferation of Th17 cells at lower concentrations (IC50:0.147 mM) compared with Th1 (IC50:0.229 mM) and Th22 (IC50:0.258 mM) and Th non-IL-22/IL-17/IFNy producing (IC50:2.14 mM) subsets. In addition, it appeared there was a paradoxical increase of HIV-1 infection levels at lower concentrations of butyrate (0.125 mM). DISCUSSION/SIGNIFICANCE OF IMPACT: The addition of butyrate to activated LP CD4 T cells decreases TCR-mediated activation in a dose-dependent manner, and butyrate acts directly on purified LP CD4 T cell populations independent of other cell populations. Butyrate differentially inhibited the proliferation of Th17, Th1, and Th22 subsets, with Th17 cells being the most sensitive to butyrate but increased the infection levels of all T helper subsets at low concentrations. Further studies are needed to determine the mechanism of butyrate’s actions on LP Th cells and the sensitivity of Th17 cells to the inhibitory effects of butyrate. These results could help direct targeted manipulation of the colonic microbiome of HIV-1 infected individuals to help resolve inflammation and limit the impact of the infection in the gut mucosa and systemically.

OBJECTIVES/GOALS: Degenerative cervical myelopathy (DCM) can lead to pain, disability, and permanent spinal cord impairment. Timely diagnosis and surgical intervention is essential to optimize functional outcomes for patients with CSM. Here, we compared patients who were admitted through clinic versus the emergency department (ED) for surgical management of DCM. METHODS/STUDY POPULATION: Patients aged ≥18 years admitted for surgery for DCM through clinic (elective cohort) were compared to a surgical cohort who were evaluated through the ED (call cohort). Basic demographics included age, gender, race, ethnicity, and insurance payor. Sociodemographic characteristics were estimated using the Social Deprivation Index (SDI) and the Area Deprivation Index (ADI) for the state of California, which were obtained through aggregated Zip Code Tabulation Area (ZCTA). Cervical MRI was reviewed to assess severity of spinal cord compression. Other outcomes included number of motion segments operated on, functional outcome using the Nurick classification, length of stay (LOS), disposition, and 30-day reoperation and readmission rates. RESULTS/ANTICIPATED RESULTS: From 2015 to 2021, 327 DCM patients received surgery (227 Elective Cohort, 100 Call Cohort). Elective cohort was mainly female (48.0 vs 30.0%, p=0.002) and white (72.7 vs 51.0%, p=0.0001). Call cohort was mainly uninsured/covered by Medicare/Medicaid (78.0 vs 67.0%, p=0.04), had higher SDI (68.0 vs 56.2, p=0.0003), ADI (7.9 vs 7.2, p=0.009), and cervical cord compression on MRI (78.0 vs 42.3% Grade III, p DISCUSSION/SIGNIFICANCE: Compared to DCM patients undergoing elective surgery, those admitted through the ED were more likely to be male, non-White, and socioeconomically disadvantaged, as measured by SDI and ADI. Postoperative outcomes were less favorable for these patients, including longer hospital stay, discharge disposition, and less Nurick grading improvement.

OBJECTIVES/GOALS: The primarygoal is to understand the challenges and barriers associated with the procurement of innovative technologies.Specifically, our research will answer the following question: what are the minimal requirements for a startup’s solution to beprocuredby anOntariohealthcare institution? METHODS/STUDY POPULATION: Participants will include procurement professionals at startups, healthcare institutions, and procurement facilitating agencies. Semi-structured interviews will be conducted in order to understand different procurement pathways and the possible procurement related gaps or barriers that startups face. Through qualitative ethnographic methods, participant interviews will characterize existing relationships and examine the rationale behind startup procurement decision-making. Data collection will include recordings, verbatim transcripts, and researcher field notes. Through inductive qualitative analysis, the data will be examined to build an intervention to assist in startup procurement. RESULTS/ANTICIPATED RESULTS: Our investigation will yield insight into expectations between hospital procurement requirements and startup procurement. The qualitative analysis will identify targets for engagement, and appropriate actors that can bridge gaps. Our results will identify pathways for procurement and the minimal procurement requirements to aid startup procurement planning. Our research will support innovators by delivering an intervention that will enable easier implementation of market ready solutions in a Canadian context. In line with principles from the National Center for Advancing Translational Sciences, this research can be used towards enhancing efficiency, speed of translation, and innovation. DISCUSSION/SIGNIFICANCE: We will contextualize the needs of start-ups and empower them to understand their procurement ecosystem. Facilitating better navigation of the procurement space allows for innovators to present solutions that healthcare organizations can adopt, resulting in improved clinical and patient outcomes.

OBJECTIVES/GOALS: The goal of U-LINK is to bring together diverse scholars from multiple disciplines to address complex and challenging problems in healthcare, climate change, social equity, and community, through innovative team science, and aligned with the University of Miami’s strategic plans. METHODS/STUDY POPULATION: The U-LINK program has supported developmental and implementation projects through a competitive selection process. Developmental funding was intended for teams to develop and refine ideas and to become established as an effective team. Additional funding was provided to teams to advance their projects by conducting data collection and feasibility testing. In addition, U-LINK has supported fellowship for pre-doctoral and affiliated doctoral trainees. Team science training was provided to all teams through didactic lectures and hands-on training. Teams were tracked longitudinally by using surveys and bibliometrics to measure success and impact including scholarly output and follow on funding. Network analysis was performed to analyze research collaboration networks before and after ULINK funding. RESULTS/ANTICIPATED RESULTS: U-LINK has funded pilot programs and initial phases for 57 projects and 13 fellowships in the last three years. Over 400 individuals on teams from 16 schools/academic units collaborated on these projects on topics such as resilience, climate change, social equity and societal challenges, health, and impact of recent legislation on LGBTQ+ community. While data collection and analysis are ongoing, initial results show successful outcomes from U-LINK projects including publications and $29.5m in follow-on external funding. We anticipate network analysis to demonstrate increased and continued multi-disciplinary collaborations among U-LINK teams through co-authorship networks and increase in collaborative grants being submitted and/or funded. DISCUSSION/SIGNIFICANCE: The University of Miami’s U-LINK program has demonstrated success in forming interdisciplinary teams that have produced real-world solutions to complex problems by harnessing the inherent diversity and strength across UM’s programs.