ABSTRACT IMPACT: This work may provide new targets for vaccine and immunotherapeutic development against MRSA infections. OBJECTIVES/GOALS: Staphylococcus aureus is the leading cause of skin and skin structure infection (SSSI), a primary portal of entry for invasive infection. Patients with SA SSSI have a high 1-year recurrence. We have shown innate memory protects mice against SA SSSI. The goal of this project is to determine epigenetic mechanisms of protective memory against SA SSSI. METHODS/STUDY POPULATION: We have shown macrophages (Mf) afford protective memory against recurrent SA SSSI in mice. Priming by prior infection reduced skin lesion size and MRSA burden, which correlated with increased Mf in abscesses and lymph nodes. Priming potentiated the opsonophagocytic killing of SA by bone-marrow derived Mf (BMDM) in vitro, and their adoptive transfer into naive skin afforded protective efficacy in vivo. Here, we investigated epigenetic mechanisms of anti-SA efficacy in BMDMs. BMDM from naive (uninfected) or primed (SA SSSI) wild-type C57Bl/6 mice were cultured ex vivo. DNA from BMDM groups were isolated and analyzed for methylation changes using reduced representation bisulfite sequencing (RRBS). Pathway analyses of methylation changes were determined with Panther. RESULTS/ANTICIPATED RESULTS: Present findings indicate the protective memory afforded by BMDM was mediated by epigenetic modifications of the DNA. Using RRBS, we profiled differentially methylated regions (DMR) in DNA from naive vs. primed BMDM. Primed BMDM exhibited significantly different DMRs as compared to naive BMDM. Proximity to known genes were mapped using GREAT. Pathway analyses revealed DMRs predominant in genes integral to immune modulation, such as integrin signaling, cytokine/chemokine networks, and growth regulation. For example, SA-primed BMDM were hypermethylated proximate to GIMAP8 versus naive BMDM, suggesting repression of this protein. Gimap family ligands are small GTPase immune-associated proteins expressed in immune cells known to regulate macrophage lysosomal fusion during parasite infection. DISCUSSION/SIGNIFICANCE OF FINDINGS: These findings reveal epigenetic mechanisms of macrophage innate memory against recurrent MRSA infection. Functional testing of these genes in response to SA infection is needed to confirm their protective role. These insights may provide new targets for vaccine and immunotherapeutic development against MRSA.

OBJECTIVES/SPECIFIC AIMS: We aimed to explore the impact of leadership training on student’s abilities to work in interprofessional research teams successfully. The Translational Research Design and Interprofessional Skills Development Course (shortened, Interprofessional Research Design) brings together students from different disciplines (science & medicine) and education tracks (PhD, MD, MD/PhD training) in a seven-week course to learn interprofessional collaborative skills and leadership styles that support success in translational research environments, while undertaking a research grant writing project. Part of the course involves a two-day leadership training workshop (12 hours) with the goal of understanding leadership styles and how to develop productive working relationships with team members to help students work more effectively in high-performance, interprofessional team environments. The course incorporates personality testing to develop self-awareness, with various exercises meant to build empathy, as well as knowledge of project management and effective leadership. METHODS/STUDY POPULATION: Nine teams of 32 students (23 MD; 9 Ph.D.) who took part in the Interprofessional Research Design course in 2017 and 2018 were required to write a reflective essay at the end of the course. We used an inductive thematic analysis to evaluate the essays. Reflective essays were coded openly by one study member. Codes were rationalized; then codes were collaboratively developed into themes by the study authors. We identified issues of integration between student groups that functioned well together and those that did not. Reflective writing responses were grouped into overall positive experiences and negative experiences. RESULTS/ANTICIPATED RESULTS: Seven of the nine teams collectively described their experiences positively. Themes related to positive team experience were “empathizing with group members”, “sophisticated communication” and “collaborative workflow/styles.” We found that those who had a positive experience utilized knowledge and skills learned during leadership training to better understand and communicate with their teammates leading to a more collaborative and dynamic workflow. These groups had higher degrees of communication both between their task assignments and within task completion periods. They also showed more awareness of others’ needs in work and communication styles. For those that had a negative experience, themes were related to “basic communication”, “poor integration” and “theory-practice gap of leadership training.” Those who struggled showed much less in- and between-task communication and showed an inability to address the personal needs of other members in communication and workflow (while still often being able to identify them). DISCUSSION/SIGNIFICANCE OF IMPACT: These findings demonstrate the usefulness of leadership training that facilitates student self-awareness and empathy, as well as effective communication, leading to collaborative high-functioning interprofessional teams. Further work incorporating conflict management and exercises to overcome the theory/practice gap of leadership and teamwork training are recommended.

OBJECTIVES/GOALS: The goal of this project was to conduct a preliminary assessment of in vivo feasibility early on in the drug-discovery process in an effort to expedite the translation of novel drug scaffolds to potential clinical candidates. The data gathered in this study will be used to direct analog synthesis of our current lead compounds through rational drug design. METHODS/STUDY POPULATION: Based on virtual and physical high-throughput screening efforts and subsequent similarity searching, we identified a set of potent and selective spermine oxidase (SMOX) inhibitors adhering to a common structural scaffold. In order to address potential barriers to in vivo use, we then conducted a robust optimization analysis in an effort to identify analogs with improved drug-like characteristics. Docking simulations to predict binding were performed and visualized using molecular modeling software (MOE and PyMol). ADMET properties were calculated using a variety of software resources including SwissADME and CDD Vault. RESULTS/ANTICIPATED RESULTS: Through these optimization efforts, we were able to successfully identify analogs with improved drug-like characteristics, including increases in predicted CNS penetration, isosteric replacement of metabolically labile functional groups, increased lipophilicity, and elimination of structural attributes suggestive of off-target activity. Analogs were ranked according to predicted binding and properties of in vivo feasibility. Compounds achieving the highest scores were then selected as scaffolds to guide analog synthesis. DISCUSSION/SIGNIFICANCE: Despite evidence implicating induction of SMOX as a mechanism contributing to neuronal pathology, the lack of potent and selective inhibitors with profiles conducive for in vivo use has significantly impeded clinical investigation of this target. In this presentation, rational drug design focusing on translational optimization will be discussed.

Pragmatic trials aim to generate timely evidence while ensuring feasibility, minimizing practice burden, and maintaining real-world conditions. We conducted rapid-cycle qualitative research in the preimplementation period of a trial evaluating a community paramedic program to shorten and prevent hospitalizations. Between December 2021 and March 2022, interviews (n = 30) and presentations/discussions (n = 17) were conducted with clinical and administrative stakeholders. Two investigators analyzed interview and presentation data to identify potential trial challenges, and team reflections were used to develop responsive strategies. Solutions were implemented prior to the commencement of trial enrollment and were aimed at bolstering feasibility and building ongoing practice feedback loops.

OBJECTIVES/GOALS: The goals of this project are to: i) investigate the cargo such as immune mediators (cytokines) and small non coding RNAs (sncRNAs) of EVs derived from nasopharyngeal secretions (NPS) of children with episodes of viral infections and exposed to SHTS; ii) test the biological activity of EVs released from upper airway mucosa on target/recipient lung cells METHODS/STUDY POPULATION: EVs were isolated from ten NPS samples of children with episodes of acute respiratory infections. EVs were characterized by particle sizing (size and concentration), EV markers, and protein arrays for interferons, cytokines, and other immune mediators content. RNA was extracted from ten samples of NPS-derived EVs by column for next generation high throughput sequencing (NGS) to identify sncRNAs in EVs. In studies currently in progress, EVs will be isolated from RSV-infected human nose organoids (HNOs) cells in air liquid interface (ALI) culture, with or without pre-exposure to tobacco smoke. EVs will be then tested for antiviral activity on recipient RSV-infected lower airway cells. Viral titers will be measured in recipient infected lung cells. RESULTS/ANTICIPATED RESULTS: We isolated EVs from NPS samples and confirmed by immunoblot EV markers CD63 and Alix. The average size of NPS-derived EVs of virus positive and negative patients was 170 nm and 145 nm, respectively. We determined the particles number of EVs, concentrations of IFN-βand IFN-λin NPS and NPS-derived EVs of these children. While IFN-βlevels were below the limit of detection in both NPS and NPS-derived EVs of all children, IFN-λwas detected in NPS and NPS-derived EVs from infected patients, except for the two patients with no viral infections. We extracted RNA from control-, virus infected- and/or SHTS- EVs and found that piR-36511, piR-40926, piR-49645, piR-32679 and piR- 53263 were all significantly enriched in EVs derived from NPS of children exposed to TS compared to those not exposed. DISCUSSION/SIGNIFICANCE: RSV leads to approximately 22,000 hospitalizations of children due to second-hand smoke. A vaccine is not currently available for RSV infection. EVs represent a novel translational approach to target undruggable. Airway mucosa EVs in viral respiratory infection function as antiviral messengers and tobacco smoke impairs the EV antiviral activity.

ABSTRACT IMPACT: Summarize the burden of diabetes comorbidities and its impact in healthcare utilization in Puerto Rico OBJECTIVES/GOALS: To estimate the prevalence of common comorbidities and describe the healthcare utilization patterns in patients with type 2 diabetes mellitus (T2DM) in Puerto Rico. METHODS/STUDY POPULATION: This is a descriptive study using healthcare claims data from patients with T2DM (based on ICD-9 diagnosis code) from most public and private healthcare insurance companies providing services in Puerto Rico in 2013 (representing more than 90% of insured population). Descriptive analyses by age, sex, type of insurance, health region, and type of medical encounter were done using frequency and percent for categorical data or means or median (with corresponding standard deviation or interquartile range) for continuous variables RESULTS/ANTICIPATED RESULTS: A total of 3,100,636 claims were identified from 485,866 adult patients with T2DM. Most patients were women (276,400; 57%), older than 65 years (235,390; 48%), from the Puerto Rico health regions of Caguas (79,604; 16%), Metro (66,280; 14%), or Bayamon (62,673; 13%) with private health insurance (371,806; 77%). The number of claims per patient ranged from 1 to 339. A mean of 6.3 claims (SD ±9.99) and a median of 3 claims (Q1 1- Q3 8) per subject were identified. Most (74%) were related to the diagnosis of diabetes (1,829,2015; 59%) or to cardiovascular diseases (458,219; 15%) and associated to outpatient services (2,722,727; 88%). The most prevalent comorbidities were hypertension (235,277; 48%), hyperlipidemia (197,449; 41%), neuropathy (100,471; 21%); renal disease (71,517; 15%), and retinopathy (61,837; 13%) DISCUSSION/SIGNIFICANCE OF FINDINGS: A high prevalence of comorbidities and use of healthcare services were identified in patients with T2DM, especially in older adults. Most comorbidities were due to diabetes-related conditions, highlighting the importance of early diagnosis and adequate management of T2DM patients to avoid preventable burden to the patient and the healthcare system

OBJECTIVES/GOALS: Septic shock is a lethal condition. Research suggests that overall sepsis mortality varies by race, but less is known about demographic differences in septic shock mortality. Our objectives were to describe the septic shock population using a large, state-wide data repository and identify demographic predictors of septic shock mortality. METHODS/STUDY POPULATION: This was a retrospective review of patients with septic shock in the One Florida Data Trust from 2012-2018. Patients were classified as having septic shock if they received vasopressors and had either 1) an ICD-9 or 10 code for septic shock or 2) an ICD-9 or 10 code for infection and an ICD-9 or 10 code for organ dysfunction. Demographic data and place of residence prior to admission was collected. The primary outcome was 90 day mortality. T-test and chi-square tests were used to test association of individual predictors and mortality. Multiple logistic regression was used to identify predictors of mortality after adjustment for other variables. Level of significance was set at 0.05. SAS v9.4 (Cary, NC) was used for analyses. RESULTS/ANTICIPATED RESULTS: There were 11,790 patients with septic shock. The mean(SD) age was 61(16) years. With regard to race/ethnicity 66% identified as white, 27% as black, 3.7% as Hispanic, and 3.5% as other races (non-white, non-black, non-Hispanic). Most came from home (57%). Overall, 39% died. Mortality varied by race (p<0.01): white 39%, black 39%, Hispanic 31%, other races 51%. In the logistic regression model, age, race, and residence were significant predictors of mortality, after adjustment for other variables. Each additional year of age had a 2.7% increased odds of mortality (OR 1.03; 95% CI 1.02-1.03; p<0.01). Compared to white patients, odds of death were 1.6 times higher for other races (95% CI 1.3-2.0; p <0.01) and non-significantly higher for black patients (OR 1.1; 95% CI 1.0-1.2; p = 0.05). Compared to those from home, odds of death were highest for those from a skilled nursing facility (OR 1.5; p<0.01). DISCUSSION/SIGNIFICANCE OF IMPACT: Patients who identified as other races had increased mortality from septic shock compared to white patients after adjusting for other variables. Septic shock mortality also increased with age and varied by residence. Further analyses are needed to examine racial disparities and control for comorbidities, severity of illness, and aspects of resuscitation. CONFLICT OF INTEREST DESCRIPTION: The authors report no conflicts of interest, except for Dr. Fernandez, who reports personal payment from Physio-Control, Inc. for speaker fees.

Long-term sequelae of severe acute respiratory coronavirus-2 (SARS-CoV-2) infection may include increased incidence of diabetes. Here we describe the temporal relationship between new type 2 diabetes and SARS-CoV-2 infection in a nationwide database. We found that while the proportion of newly diagnosed type 2 diabetes increased during the acute period of SARS-CoV-2 infection, the mean proportion of new diabetes cases in the 6 months post-infection was about 83% lower than the 6 months preinfection. These results underscore the need for further investigation to understand the timing of new diabetes after COVID-19, etiology, screening, and treatment strategies.

OBJECTIVES/GOALS: The Association of Clinical and Translational Science (ACTS) chartered a Diversity, Equity, and Inclusion (DEI) Committee to prioritize activities to support, develop and report on metrics for measuring progress toward DEI goals. This poster aims to describe the formative process toward prioritizing DEI in society’s efforts. METHODS/STUDY POPULATION: In 2021, the ACTS chartered the DEI Committee. Two ACTS Board of Directors members chair the committee, and members represent 10 academic institutions committed to prioritizing DEI. Members participated in a Human-Centered Design process to develop the committee’s mission, goals, and activities. The committee determined areas of opportunity for ACTS to increase DEI by identifying challenges to support DEI in clinical and translational science. The chairs facilitated a discussion using Mural to foster an interactive strategy to engage members in conversations that respected individual experiences, promoted a discussion of actions that ACTS as a society, and determining metrics for measuring DEI. RESULTS/ANTICIPATED RESULTS: We plan to present the committee’s efforts to create the ACTS mission statement and strategies to priority to DEI. We will describe current and future activities to engage historically underrepresented members and academic institutions in ACTS programs. Based on the committee’s work, ACTS has taken systematic approaches toward social justice and is beginning to determine new ways of engaging members. DISCUSSION/SIGNIFICANCE: Scientific societies that prioritize DEI increases equity and belonging across members. ACTS is at the forefront of advocacy, policy, and social justice effort. The DEI committee is positioned to aid ACTS in increasing DEI across the clinical and translational science spectrum.

OBJECTIVES/SPECIFIC AIMS: The current study is the first investigation of frontal alpha asymmetry in distressed violent (DV) and distressed nonviolent (DNV) partners during a placebo-controlled alcohol administration and emotion-regulation study. Because this is the first study of the pharmacological effects of alcohol on FAA, the first portion of the study was conducted to characterize alcohol effects in DV and DNV partners during the baseline condition. The subsequent portions of the study were conducted to characterize the effects of alcohol and evocative stimuli on FAA in DV and DNV partners. We hypothesized that DV partners would demonstrated greater left frontal alpha asymmetry when intoxicated and viewing evocative partner stimuli than DNV partners. Lastly, we attempted to replicate previous research that has found associations between baseline measures of FAA and the State-Trait Anger Expression Inventory – 2 (Spielberger, 1999) subscales of Trait Anger, Anger Expression-Out, Anger Expression-In, Anger Control-Out, Anger Control-In (Hewig, Hagemann, Seifert, Naumann, & Bartussek, 2004). METHODS/STUDY POPULATION: Partners in the present study were drawn from a larger study investigating over-arousal as a mechanism between alcohol use and intimate partner violence (AA022367). Couples were recruited from the community via radio, television and newspaper advertisements, and eligibility screening occurred at the couple level. Participants included in the present analysis were 23 DV partners (12 female, 11 male), and 15 DNV partners (7 female, 9 male). The mean age of the sample was 32 (SD 4.8 years, range 23-40 years). Data from two DV partners were not included in the analyses of the FAA in the emotion-regulation tasks due to movement artifacts during the alcohol condition leaving too little data for analysis. RESULTS/ANTICIPATED RESULTS: The expected beverage by couple type interaction did not reach significance [F (1, 36) = 3.93, p = .055], but the between-subjects effects of couple type revealed a significant difference [F (1, 36) = 4.425, p = .042]. Contrary to our hypothesis, however, these results suggest that under conditions of alcohol, DV partners evidenced significantly greater relative right frontal alpha power asymmetry whereas DNV partners evidenced greater relative left frontal alpha power asymmetry. Although there was no significant between-subjects effect, there was a nearly significant interaction between beverage type and emotion regulation condition [F = (1, 36) = 4.032, p = .052] and a significant main effect of emotion regulation condition [F (1, 36) = 7.579, p = .009]. It appears that asking the participants to “not react” to their partners’ evocative stimuli caused significantly greater right frontal alpha asymmetry. Because intimate partner violence is best understood in the context of conflict between two partners, we also examined partner-reported experiences of anger as predictors of DV participant’s FAA. The model as a whole predicted 67.4% of the variance in DV partner FAA, R squared change =.674, F Change (5, 15) = 6.21, p = .003. Three anger experience scales were statistically significant. The partner Anger Control-Out (B = -1.23, p =.001) scale recorded a higher standardized beta value and accounted for 40% of the variance in this model. Anger Control-In (B = .63, p = .022) accounted for 14% of the variance in the model, and Anger Expression-Out scale (B = .57, p = .024) accounted for 13.7% of the variance in the model. DISCUSSION/SIGNIFICANCE OF IMPACT: The current study is the first pharmacological study of the effects of alcohol on frontal alpha asymmetry in distressed violent and distressed nonviolent partners. Contrary to our hypothesis, under acute alcohol intoxication during the baseline condition, DV partners exhibited significantly greater relative right FAA compared to DNV partners who exhibited significantly greater relative left FAA. Because intimate partner violence is best understood in the context of couple conflict, we examined the ability of partners’ anger experiences to predict DV and DNV partners’ FAA, and a very interesting pattern emerged among our DV participants and their partners. The anger experiences of our DV participants’ partners accounted for 67% of the variance in the FAA of our DV participants when they were intoxicated and viewing evocative stimuli.

OBJECTIVES/GOALS: The Association for Clinical and Translational Science (ACTS) offers important inter-institutional communities through Special Interest Groups (SIGs). New to the SIG ecosystem is the Justice, Equity, Diversity, and Inclusion (JEDI) SIG. METHODS/STUDY POPULATION: Formed in 2022, the JEDI SIG is a growing community of individuals seeking to develop papers and products relating to improving health equity and workforce diversity within the Translational Science Community. Within this context, diversity is broadly defined to include groups at a negative power differential within US society. RESULTS/ANTICIPATED RESULTS: Comprised of 54 individuals spanning 29 institutions as of our first meeting in October 2022, the JEDI SIG is a welcoming group for ACTS members looking to make a difference. We have developed working groups to develop SIG-directed projects, and we look forward to expanding our activities in the future. DISCUSSION/SIGNIFICANCE: In this poster we will share key information about the new JEDI SIG including monthly meeting times, current activities, and ideas for future work.

ABSTRACT IMPACT: The described framework will enable other sites with a well-defined apparatus for enabling the secondary analysis of EHR data for research through education, team science, and resource consolidation. OBJECTIVES/GOALS: EHR’s potential to improve healthcare outcomes extends far beyond the clinic. This vast repository of clinical insights has dramatic potential for biomedical research. To enhance accessibility for busy clinicians and underserved populations, we describe a framework for interfacing with EHR locally and through national network participation. METHODS/STUDY POPULATION: The Institutional Development Award (IDeA) program, which began in 1993, broadens NIH funding’s geographic distribution for biomedical research. Included in this is the IDeA Networks for Clinical and Translational Research, which focuses on enhancing clinical and translational science across a network of IDeA-states with traditionally underserved communities and rural providers. A prior survey of the needs and capabilities of IDeA-CTR centers identified the need for improved research support. Based on our annual member survey we developed a process for supporting distributed research projects across the GP-CTR. NIH also recently made a funding announcement for the IDeA-CTR community identifying EHR research as a major priority in responding to the COVID-19 pandemic. RESULTS/ANTICIPATED RESULTS: Results from site interviews and member surveys show a clear need for dedicated resources to navigate the process of EHR-derived research. Most described a different set of requirements for increasing accessibility to EHR for research and a strong desire to participate in research networks. Local investigators cited a lack of tools, educational materials, and accessibility. Initial efforts demonstrate strong research questions but limited technical, statistical, and terminological capabilities to succeed. In response, a pipeline for team science and promotion of projects from local phenotypes to national studies. We created a facilitator training program to expand the number of facilitators (n=22), quarterly training for investigators (n=104), and ongoing efforts to advance COVID-19 research. DISCUSSION/SIGNIFICANCE OF FINDINGS: As evidenced in the expanding number of EHR-based research networks there is a need for a system to promote project development and best practices. The proposed model promotes education, resource sharing, and team formation to advance clinical questions from the idea stage toward national research network participation.

OBJECTIVES/SPECIFIC AIMS: We sought to solicit and synthesize stakeholders’ ideas for how the Advance-CTR program can best increase the quality and quality of clinical and translational research in Rhode Island, and to apply these findings to address barriers and strengthen research capabilities across our partner institutions. METHODS/STUDY POPULATION: We utilized a Group Concept Mapping approach, involving university and Institution-based researchers and administrators. The process was conducted using the web-based concept mapping application CS Global Max (Concept Systems, Inc). Respondents were asked to provide their best ideas for promoting clinical and translational research in RI. These ideas were then organized by our project team into a set of unique items for consideration by attendees of an Advance-CTR retreat. Participants were tasked with sorting these ideas by theme (cluster), and were also asked to rate each idea according its importance and feasibility. Using the online software, these clusters and ratings were analyzed to identify key themes and to explore differences among sub-groups. RESULTS/ANTICIPATED RESULTS: The Group Concept Mapping exercise yielded 150 statements that were edited down to 78 unique ideas, and clustered into nine themes (e.g., institutional collaboration, training). Fifty-seven retreat participants completed the sorting and rating tasks of the concept mapping exercise. Overall, ideas rated as highly important and highly feasible included “providing seed grants to encourage new collaborations across basic science,” and “connecting researchers with common interests.” Top rated items varied across institutions and according to respondent demographics, allowing us to consider the unique issues relevant to particular groups. Relative rankings of clusters across groups revealed notable differences, such as higher importance placed on community engagement among administrators as compared with researchers, and differences in needs for internal support for research between universities. DISCUSSION/SIGNIFICANCE OF IMPACT: Group Concept Mapping was an effective and insightful participatory approach to engage our program’s stakeholders in developing ideas and identifying challenges to enhancing clinical and translational research in Rhode Island. Our results have implications for project decision-making and initiatives to facilitate translational research in RI. Thus, results have been presented to the Advance-CTR community via webinar, as well as Advance-CTR project leadership and advisory committees.