OBJECTIVES/GOALS: Our goal is to develop a cost-effective approach for precision medicine treatment by providing computational predictions for new uses of currently available FDA approved, and experimental drugs for NSCLC. METHODS/STUDY POPULATION: Cell Lines: A549 (ATCC- CCL-185) Human epithelial Lung Carcinoma cells, H1792 (ATCC-CRL-5895) Human Lung Carcinoma cells. In Vitro Cytotoxicity Assay: A Vybrant® MTT Cell Proliferation Assay was used. Colony Formation Assay: NCI-H1792, A549 cells were seeded at a density of 500 cells/ dish, then treated with ARS-1620, Osimertinib. The Computational Analysis of Novel Drug Opportunities (CANDO):  Herein, we employed the bioanalytic docking (BANDOCK) protocol within CANDO to calculate the compound-protein interaction scores for a library of 13,218 compounds from DrugBank against a library of 5,317 protein structures from the Protein Data Bank, resulting in a proteomic interaction signature for each compound, and identified Osimertinib as the most likely EGFR/ErbB inhibitor to synergize with ARS-1620. RESULTS/ANTICIPATED RESULTS: ARS-1620 and Osimertinib in combination displays potent anti-tumor activity as evident by a decrease in cell viability with cytotoxicity assays, as well as reduced number of colonies in the colony formation assay for both A549 and H1792 cells. By using CANDO, and cross-referencing the obtained rankings with known experimental information, we have obtained drug predictions within the context of precision medicine. Our preliminary data indicates that EGFR inhibitor Osimertinib may be most structurally similar to KRAS G12C inhibitors overall, compared to other ErbB/ EGFR inhibitors. Validations with human cancer cell lines A549 and H1792 have confirmed that Osimertinib in combination with KRAS G12C inhibitor ARS-1620 may exhibit a synergistic effect in decreasing cellular proliferation and colony formation. DISCUSSION/SIGNIFICANCE: This suggests that this innovative drug combination therapy may help improve treatment outcomes for KRAS G12C(H1792) and KRASG12S(A549) mutant cancers. Cell migration and cell invasion studies in response to treatment with Osimertinib and ARS-1620 are currently ongoing.

OBJECTIVES/SPECIFIC AIMS: This case-control study aims to determine the relationships among childhood adversity, attachment style, and the likelihood of accepting or declining a referral for HV. The study will serve as a pilot to inform the power analysis of a subsequently proposed full-scale study. METHODS/STUDY POPULATION: Using a case-control study design, 25 women who decline HV referral (cases) will be compared with 25 women who accept HV referral (controls) on their exposure to childhood adversity and attachment style. Women who are eligible for the study are English-speaking mothers who have been offered HV services by Health Care Access Maryland. Surveys are administered in-person, either in the participant’s home or at another location (e.g., public library), based on participant preference. The dependent variable is participant’s verbal response to the HV referral (accept/decline). The independent variable, childhood adversity, will be measured using the Philadelphia Urban Adverse Childhood Experiences (ACEs) Survey and the Attachment Style Questionnaire (ASQ). Control variables include demographics (i.e., age, race, education, employment, housing, marital status), obstetric history (i.e., previous preterm birth, miscarriage, fetal death, infant death, abortion), and current psychosocial risk factors (i.e., history of substance use, intimate partner violence, depression). Descriptive comparisons will be done for the independent and control variables in controls versus cases. Bivariate analysis will examine associations between the odds of being a case and ACE score and ASQ score. Multivariate logistic regression models will be used to examine the relationship between ACE total and ASQ score; exposure to ACE in cases versus controls; and the odds of an avoidant and anxious attachment styles in cases versus controls. RESULTS/ANTICIPATED RESULTS: We hypothesize that (a) higher ACE scores will be positively associated with a higher level of avoidant attachment; (b) higher ACE scores will be positively associated with declining a HV referral; and (c) higher levels of avoidant attachment will be associated with declining a HV referral. DISCUSSION/SIGNIFICANCE OF IMPACT: Racial inequities in birth outcomes are pervasive and unjust. Non-Hispanic Black women experience births that result in infant mortality, fetal mortality, preterm birth, and low birth weight babies at more than double the rate of non-Hispanic White women in Baltimore and nationally. Prenatal and early childhood home visiting programs have been found to decrease maternal smoking and hypertensive disorder which are associated with PTB, reduce closely spaced births which is associated with fetal and infant death, and improve women’s long-term economic self-sufficiency, child health and social outcomes. However, as community-based programs, these services are not reaching the majority of eligible women in low-income urban settings—women who are also disproportionately burdened with poor pregnancy-related health outcomes. Considering the potential to improve outcomes, the importance of eliminating health disparities, and the national and local investment in HV services, it is vital to understand why some women are not enrolling in prenatal HV programs. The findings from this and subsequent studies will inform the translation of evidence-based HV program outreach efforts for women with complex social history. It will inform the design of enhanced outreach and engagement efforts of HV programs to more reliably engage women.

OBJECTIVES/GOALS: Waiting for psychotherapy is a major barrier to care and associated with negative outcomes. Individuals waiting for treatment may be particularly well-suited to receive low-intensity treatments (LITs), but few providers recommend LITs. We investigated provider-identified barriers to recommending LITs for patients on treatment waiting lists. METHODS/STUDY POPULATION: We recruited mental health professionals via social media and professional association listservs to participate in a brief survey. Participants were asked about their current waiting list practices and attitudes towards low-intensity resources for patients waiting for treatment. Participants were prompted to provide additional thoughts on recommending LITs for patients on waiting lists in an open-ended text box. Two members of the research team independently coded responses into themes, resolved discrepancies, and achieved total consensus. RESULTS/ANTICIPATED RESULTS: 141 mental health providers participated in the survey, and 65 (46%) provided a response to the open-ended question. The emerging themes included: Patient Barriers, Research Evidence/Efficacy, Feasibility, Patient Personal Contact, Patient Appropriateness, Liability, Systemic Problems, Trust in Programs, Downplaying Distress, Additional Resources, and Positive Attitudes. Providers were particularly concerned with giving a generalized intervention without having conducted a full evaluation or assessment with a patient. Many providers also reported concerns pertaining to the legal and ethical liability of providing LITs when a patient is not being seen face-to-face by a provider. DISCUSSION/SIGNIFICANCE: Many of the themes we identified parallel those identified in previous literature. Some barriers we identified from our providers, when thinking about integrating LITs on waiting lists, highlight the need for professional guidelines to address legal and ethical liability, as well as billing and reimbursement procedures.

OBJECTIVES/SPECIFIC AIMS: Stakeholder engagement has been proposed to help realign clinical and translational research with the needs of clinicians, patients, and policymakers. Increasingly, funders and researchers seek to partner with stakeholders to inform study design, execution and dissemination of results. Kids and families Impacting Disease through Science (KIDS) is a program of the American Academy of Pediatrics that seeks to engage youth in clinical research. United States KIDS programs participate in International Children’s Advisory Network activities. The Chicago KIDS Advisory Board program at Walter Payton College Preparatory School, a Chicago Public School, was initiated in 2015 to foster and develop interest in careers in science, research and healthcare and provide youth perspectives to academic and industry researchers on the design and development of pediatric research studies. This project engaged youth advisors in creation and evaluation of a video explaining clinical research and informed consent for Ann & Robert H. Lurie Children’s Hospital, a clinical partner of the Northwestern University Clinical and Translational Sciences Institute. METHODS/STUDY POPULATION: The Payton program advisory board sessions are 1.5hr interactive seminars held on 1–2 school days each month. During the 2016–2017 school year, students participated in 3 stakeholder sessions, led by Lurie Children’s hospital researchers, to advise development of a script, storyboards, and ultimately an animated video that informs children and families about participation in clinical research to aid in the decision-making process. Qualitative research methods were used to examine attitudes towards clinical research and assess the video on content objectives, clarity of concept, and appropriateness for a pediatric audience. Following production, students from the 2017–2018 advisory board viewed the final video and presurvey and postsurvey were administered to assess the effect of video on the comprehension of 8 key concepts of informed consent on a 5-point Likert scale. The Wilcoxon signed-rank test was used to compare median pretest and post-test ranks. Results of this analysis were reviewed in seminar and students provided written contribution to this abstract. RESULTS/ANTICIPATED RESULTS: In total, 11 Walter Payton high school students participated in video development and 27, who were naïve to development, participated in the pre and post evaluation sessions. Students ranged from Freshman to Seniors and reflected the diverse ethnic and racial background of Chicago. A positive change from pre to post-test survey was observed in all questions presented assessing comprehension of key concepts of informed consent. The median post-test ranks were statistically significantly higher than the median pre-test ranks for all questions (p<0.01 in all). DISCUSSION/SIGNIFICANCE OF IMPACT: Chicago KIDS youth advisors were engaged in all aspects of the design of the research tool and gained experience in stakeholder contribution from study design to evaluation and publication. The students will next be involved in the design of a prospective randomized study to test the efficacy of the video compared with standard recruitment and consent practices. Given the difficulty of recruiting youth for clinical trials, development of effective engagement practices in is critically important. Our findings demonstrate the feasibility of utilizing youth advisors in a public school based setting.

OBJECTIVES/SPECIFIC AIMS: The objective of this research was to investigate the effect of alcohol and evocative stimuli on heart rate variability (HRV) in partners with a history of intimate partner violence in a placebo-controlled alcohol administration study with an emotion-regulation task. METHODS/STUDY POPULATION: In total, 17 partners (9 females, 8 males) with a history of partner violence participated in a placebo-controlled alcohol administration study with an emotion-regulation task during which HRV measures were collected. In the alcohol condition, participants were administered a mixture of 100 proof vodka and cranberry juice calculated to raise their blood alcohol concentration (BAC) to 0.08%. In the placebo condition, participants consumed a volume of juice equivalent to that consumed in the alcohol condition, but without alcohol. Alcohol and placebo conditions were counter-balanced across participants as were the presentation the blocks of evocative and neutral partner stimuli. RESULTS/ANTICIPATED RESULTS: Controlling for baseline HRV, there was a significant main effect of stimuli (evocative vs. neutral partner stimuli) on HRV in intoxicated partners, F1,16=16.28, p=0.004. There was also a significant main effect of regulation on HRV under conditions acute alcohol intoxication, F1,16=23.55, p=0.001. These effects tell us that intoxicated partners experienced reduced HRV when exposed to evocative stimuli from their partners. These effects also tell us that under acute alcohol intoxication, partners were less able to regulate their emotion when exposed to evocative stimuli than when they consumed a placebo beverage. DISCUSSION/SIGNIFICANCE OF IMPACT: These results suggest that increases in intimate partner violence under acute alcohol intoxication may be the result of reduce HRV. This reduction in HRV would contribute to partners’ inability to response with adaptively in conflict when intoxicated. They also suggest that HRV may be an important target for intervention with partner with a history of intimate partner violence. One method may be Heart Rate Variability Biofeedback which has been shown to increase parasympathetic nervous system functioning, autonomic stability, and emotion regulation.

OBJECTIVES/GOALS: Innovative educational approaches and training modalities are important for training a diverse workforce in the authentic skills needed to advance all phases of clinical and translational research. Endeavors to study and develop policies that promote the translational science spectrum are steeped in value judgments. Learning how to navigate moral ambiguity and ethical reasoning enlightens our understanding of stakeholder obligations, roles and responsibilities. Ethics education can be challenging if learners are insufficiently engaged in the necessary critical reflection. In this course, decision-making in public health is informed through the analysis of the ethical issues, developing alternative courses of action and providing justification for actions taken in response to real-world dilemmas. The course is provided to students with a variety of backgrounds (science, health, policy) in a Master of Public Health degree program. Course objective were to: 1) Identify ethical issues in public health policy, practice, and research using appropriate concepts and terms; 2) Recognize the full spectrum of determinants of health and related information needed to resolve ethical conflicts in public health policy, practice, and research; 3) Present varied and complex information in written and oral formats; 4) Assess potential solutions to ethical conflicts in public health policy, practice, and research and 5) Decide ethical courses of action for public health policy, practice, and research. We adopted an open pedagogy as a guiding praxis to inform public health ethics discourse amongst our learners. In this way, learner agency was maximized to develop course materials within a generalized framework and shared with each other through the perspectives of each individual. The goal was to not only analyze complex ethical dimensions of public health issues but also gain insights into the disciplinary lenses of one’s peers. METHODS/STUDY POPULATION: Each week was divided into two sessions, a seminar and workshop. Course instructors introduce topics in a one-hour session and then allow students to decide what information is needed for a second session where the ethical issues of the topic will be discussed. Information-gathering tasks are then distributed amongst students in areas that are not their specialty, e.g. social history to be researched by learner with a biology background. The second session then involves the reporting back of background information by each student and a discussion of the ethical issues that arise. Through this process, the ability to communicate with others in different disciplines is supported, while exploring other disciplines and then engaging in ethical discussion and reasoning. Topics were introduced during the seminar session each week over the span of five weeks: 1) global public health, 2) disease prevention & control, 3) environmental & occupational public health, 4) resource allocation & priority setting and 5) research ethics. Learners were tasked with identifying the needed information to address the ethical, policy, and research aspects of the public health question(s) presented in these seminars. Students independently submitted resources they discovered to course instructors prior to the workshop. The following session began with a workshop where learners briefly presented their findings and deliberated on specific facets of the public health issue from that previous seminar while discussing a specific case. Students were assessed on their preparation (submission of identified resources), workshop presentation and participation. Research Preparation: In each seminar, the class decided what key information would be required to support the discussion at the workshop, which revolved around a relevant case study on that week’s topic. Course instructors facilitated the groups identification of material to be researched and the delegation of tasks within the group. Each student submitted a summary document (template provided) to course instructors prior to class for their area of research related to the case. Research Presentation: At the beginning of each workshop, each student was asked to present the research work to the rest of the class so that everyone has the same information for the case study discussion. These short (5-10 minutes) presentations followed the format of the preparation summary. Participation/collaboration: Both the seminar and the workshop asked students to be active learners within the class, participating in discussion, strategizing for information-gathering tasks, presenting researched material and arguments to others, and participating in case study discussion. Participation was assessed in relation to the value of the contributions made by students. RESULTS/ANTICIPATED RESULTS: The open pedagogy allowed the learners to construct the necessary materials to discuss issues with each other and develop not only a deeper understanding of the ethical dimension of public health issues but a shared understanding of each other’s disciplinary lenses. Course feedback was generally very positive, with learners either agreeing (33%) or strongly agreeing (67%) that the course was effective overall. In asking what learners liked best about the course, some indicated the “open pedagogy learning style” and “I liked the discussion format.” The positive comments mostly highlighted the discussion format. Areas for improvement noted by the learners included wanting “a longer course to cover more topics” and that the material was covered in “too short a time frame.” Other comments included that the course “was a bit disorganized” or that “the discussions were not very structured.” While the discussions by their very nature were unstructured, there is opportunity to refine this pedagogy to find right balance of learner agency. DISCUSSION/SIGNIFICANCE OF IMPACT: The goal of this teaching method was to empower the learner with the important critical thinking skills to navigate challenging ethical dilemmas in public health they may encounter in their careers. These skills include the identification of the ethical or moral conflict(s), collecting the necessary information to examine/resolve the dilemma, think creatively about the information that is unavailable and how to discuss/disseminate information to a broad constituency. This an educational model that is easily adaptable for learners working in other areas of the translational research spectrum, e.g. basic, pre-clinical, clinical and implementation sciences.

OBJECTIVES/SPECIFIC AIMS: 1. Foster a community of scholars – by centering on training and professional development programs that assist with career progression. 2. Improve heath equity for the community – by developing innovative ways to capture data, address problems, deliver solutions, and disseminate results to patients and people everywhere. 3. Facilitate interdisciplinary teams at all career stages – by providing opportunities for researchers, regardless of discipline and career stage, to engage in dialogue with others. SCOR has developed a series of programs to address each stage in the career arch. LEAD: Learn Enhance Advance Drive develops a pipeline of future leaders within UAB by engaging junior faculty and staff who are in leadership roles or will be in leadership roles soon to develop fundamental competencies. The one-year, cohort-based program enables junior faculty and staff to enhance their interpersonal skills, professional skills, and leadership skills. The program uses the Leadership Competency Model to provide the framework for the chosen topics. The K2R program provides structured activities over a 4– 5 month timeline to assist cohorts of scholars in their preparation of a specific grant application. Leveraging existing programs, like Project Panels, and offerings including a Specific Aims Workshop, K and R Writing Groups, and Mock Study Section, the SCOR extends the engagement of scholars in their ongoing research development. METHODS/STUDY POPULATION: Using quantitative methods, we have conducted surveys to measure effectiveness of weekly didactic sessions and topics. We use qualitative methods such as interviews and focus groups to better understand the relevance of the individual programs and the larger community of SCOR. RESULTS/ANTICIPATED RESULTS: N/A DISCUSSION/SIGNIFICANCE OF IMPACT: Creating a community of interdisciplinary investigators will greatly impact research at the institution.

OBJECTIVES/SPECIFIC AIMS: Tissue engineered tracheal grafts (TETG) could provide a life-saving cure for children with long segment airway defects. Computational fluid dynamics (CFD) is a novel and promising technique used to evaluate TETG performance. This pilot study examines the correlation of objective CFD simulations with subjective respiratory symptoms in a TETG large animal model. METHODS/STUDY POPULATION: Three-dimensional geometries of 1 TETG implanted sheep trachea were reconstructed from serial fluoroscopic images, allowing analysis with CFD simulations. Peak flow velocity (PFV) and peak wall shear stress (PWSS) across the graft as well as changes secondary to stenting were determined. CFD metrics were compared with respiratory symptoms seen on exam. RESULTS/ANTICIPATED RESULTS: Two weeks after implantation, the animal developed respiratory distress, which correlated with PFV and PWSS elevations. Although the intraluminal graft appearance changed minimally after dilation, PFV and PWSS decreased across the graft (4.5–0.8 m/s and 0.9–0.1 Pa, respectively). Long-term TETG stenting with dilation returned PFV and PWSS to baseline (0.8–0.3 m/s and 0.1–0.01 Pa, respectively), which correlated with immediate symptom resolution. DISCUSSION/SIGNIFICANCE OF IMPACT: CFD is a noninvasive modality, which allows the evaluation of airflow metrics of symptomatic TETG recipients. This diagnostic tool will permit planned interventions and graft design optimization.

OBJECTIVES/GOALS: 1) Characterize racial differences in congestive heart failure care delivery. 2) Examine the extent to which specific clinical roles were associated with improved care outcomes (i.e., hospitalizations, readmissions, days between readmissions, and charges) of African Americans (AA) with CHF. METHODS/STUDY POPULATION: EMR data was extracted from the Arkansas Clinical Data Repository (AR-CDR) on patients (ages 18-105) who received care between January 1, 2014 and December 31, 2021. Variables included age, sex, race, ethnicity, rurality, clinical diagnosis, morbidities, medical history, medications, heart failure phenotypes, and care delivery team composition. Binomial logistic regression ascertained the effects of these variables on patient’s care outcomes. A Mann Whitney-U test identified racial differences in outcomes. Psychometrically, classical test theory and item response theory assessed items for the risk surveillance tool. RESULTS/ANTICIPATED RESULTS: The study identified 5,962 CHF patients who generated 80,921 care encounters. The results revealed the disproportionate impact of CHF prevalence, hospitalizations, and readmissions on AAs. AAs had a significantly higher number of hospitalizations (i.e., 50% more) than Caucasians. Specific clinical roles (i.e., MDs, RNs, Care Managers) were consistently associated with 30% or greater decrease in odds of hospitalization and readmission, even when stratified by heart failure phenotype. Classical test theory results (e.g., Cronbach’s alpha; 0.88) indicated the set of items on the risk surveillance tool accurately reflect a patient risk for improved outcomes. DISCUSSION/SIGNIFICANCE: The findings stimulate the need for 1) EHR-based tools that manage care delivery equity and 2) investigations of specific clinical roles in risk stratifying and operationalizing the care plans of AAs, advancing formal access-to-care frameworks by ensuring access to clinical roles that are associated with improved outcomes.

OBJECTIVES/SPECIFIC AIMS: Current translational research moves beyond bench to bedside and includes translating scientific evidence to clinical practice and into the community settings (T1-T5). This progression is dynamic, involving patient-physician, community, and academic organizational structures and translational strategies. However, basic and clinician scientists are often unprepared and/or ill equipped to successfully conduct community-engaged research which may aid in more efficient translation of their research findings. The recognized need for such training was the impetus for our course which was originally designed and implemented through the innovative and sustainable joint academic-community partnerships of Morehouse School of Medicine and Emory University with the support of Georgia Institute of Technology. Since that time the course has evolved with the recently added partner, University of Georgia. METHODS/STUDY POPULATION: Initially developed and implemented in 2008, the course continues through the Georgia Clinical and Translational Science Alliance, Community Engagement and Research Program (GaCTSA/CERP), a Clinical Translational Science Award (CTSA) (UL1TR002378). The course is an introduction to community-engaged research concepts/methods. This includes behavioral science; community engagement principles; clinical translational research partnerships; and strategies in planning, implementing, evaluating, and disseminating community-engaged research to address health disparities. The course is open to the four GaCTSA academic institutions’ faculty, MD, PhD, MS in Clinical Research, and the Graduate Certificate in Translational Science students. RESULTS/ANTICIPATED RESULTS: Students received scholarly and hands-on training in community engaged research through faculty- and community member-led didactic lectures/interactions, team science activities, and a final assignment involving work with a community-based organization. From 2008-2017 over 230 students have matriculated through this course and many are now involved in community-engaged translational research. Most students in the class were MD/PhD students (33%), however 21% were junior faculty, attending physicians (21%), or fellows/residents/ postdocs (15%). Evaluations over the years indicate that most students were unware of Community-Based Participatory or community-engagement strategies for conducting translational research. DISCUSSION/SIGNIFICANCE OF IMPACT: Effective application of community-engaged translational research requires essential skills training to facilitate the translational research paradigm. Translational researchers, at any stage, will benefit from understanding the entire translational research process and the importance of quickly bringing research advances to patients and the community.

ABSTRACT IMPACT: The Hispanic Alliance for Clinical and Translational Research Professional Development Core (PDC) will contribute to the improvement of the health of an increasing US Hispanic population, by supporting and training a new cadre of Hispanic/Latino CTR researchers and community leaders that understand this population’s prevalent health needs. OBJECTIVES/GOALS: To use the Professional Development Core (PDC) of the Hispanic Alliance for Clinical and Translational Research (Alliance) as a hub that coordinates training, mentoring programs, and grant support to address the need for more underrepresented minorities (URM) in clinical and translational research and mentoring. METHODS/STUDY POPULATION: PDC will: (1). Coordinate and offer an effective educational program based for new and mid-career researchers to address the gaps in research competencies on Hispanic/Latino health and healthcare through web-based asynchronous distance training, enhanced with face-to-face interactions. (2). Establish a robust mentoring program to address the mentoring gap for URM faculty by developing mentorship skills of faculty and researchers through a variety of resources, and offering protected time to mentor-mentee teams. (3). Design and implement a tailor-made curriculum to train scientists and community partners jointly, enabling them to carry out multidisciplinary research responsive to the Hispanic/Latino community health’s needs. RESULTS/ANTICIPATED RESULTS: From 2010 to 2019 the PDC supported over 1,000 researchers and faculty and provided 52 activities over the 9 years. PDC-supported researchers submitted 56 proposals and 21 grants (37.5.%) were awarded, for a total of $2, 225,751.00, and to published 94 peer-review papers. We expect that through Alliance PDC will sponsor at least 20 new trainees/mentees in Clinical and Translational Research (CTR), 20 new certified mentors, a continuous support program, and an increase of 30% in the scientific productivity (e.g., grants submission and peer-reviewed publications) of the Hispanic CTRs in Puerto Rico and the establishment of long-term links with the Hispanic community in Puerto Rico and across the United States to address its health needs. DISCUSSION/SIGNIFICANCE OF FINDINGS: The PDC programs are significant in addressing the need for qualified researchers and mentors that understand, have the know-how, and are interested in addressing the health needs of a growing USA Hispanic medically underserved population.

OBJECTIVES/SPECIFIC AIMS: The specific objectives of this project are (1) identify, test, and validate the parameters for a simplified NLOM imaging probe that will provide specific research and point-of-care information on biofilm presence, therapeutic need and response of individual wounds to treatment. (2) Identify specific proteomic and metabolomic biomarkers of (i) wound susceptibility to infection, (ii) wound response to the most commonly used antibacterial measures in wounds, and (iii) establish criteria for more effective interventions. METHODS/STUDY POPULATION: First, optimal use parameters for NLOM including illumination, field of view, focal length, linear Versus concentric image acquisition, detection and filter wavelengths were identified. Parameters for evaluation included ease and speed of imaging, ability to map diagnostic criteria. Next, using the optimised NLOM imaging modality in bacterial biofilm isolates and subsequently a rabbit ear model of biofilm wound infection, proteomic and metabolomic biomarkers of susceptibility to infection were identified. The effects of 2 standard debridement and anti-infective treatments, polyvidone-iodine solution or cetrimide 15%+ chlorhexidine gluconate 1.5% were mapped in situ for up to 10 days using the NLOM probe. RESULTS/ANTICIPATED RESULTS: Using the novel custom NLOM probe, high resolution mapping of wound biofilm infection, as well as the underlying tissue was performed throughout the onset, development, treatment, and resolution of wound biofilm infection. Specific microbiological, microstructural, oxygenation, and pH parameters were mapped at defined surface and subsurface locations and time-points. Findings included the determination that some standard antimicrobial formulations provide a supportive environment for wound infection, and that micro-channels within the biofilm and their interface with the tissues serve as an important predictor and indicator of wound infection establishment, progression, and response. DISCUSSION/SIGNIFICANCE OF IMPACT: The novel multimodality in vivo NLOM imaging approach establishes an important tool for earlier and more specific diagnosis of wound infection risk, virulence, and invasiveness along with markers of successful treatment, and a simple clinical imaging tool for improving wound infection prevention and treatment.

OBJECTIVES/GOALS: Promoting Academics and Community Engagement (PACE) Dissemination grants are funding opportunities via the Michigan Institute for Clinical and Health Research Community Engagement program designed to support the science of clinical and translational research demonstrating community impact. METHODS/STUDY POPULATION: Two statewide funding announcements over a 3-month period were issued to over 2,000 academic and community partners. Proposals were required to meet the following criteria: 1) data collection from an academic and community partnered research project completed; 2) research findings analyzed; and 3) a community-focused dissemination plan developed. Projects were funded up to $5000. RESULTS/ANTICIPATED RESULTS: Six PACE dissemination grants with community-focused plans were funded spanning Southeast, Western, and the Upper Peninsula regions of Michigan, as well as statewide. Examples of funded projects topics areas include the following: firearm safety; housing discrimination; opioid misuse; suicide prevention; and youth mental health. Community dissemination activities include: a) presenting at a community town hall; b) writing a pamphlet for community use; c) creating artwork installation showcase for community display; d) storytelling through community channels; and e) designing and implementing a local social media campaign. DISCUSSION/SIGNIFICANCE: Traditional funding mechanisms often do not provide resources to disseminate research findings with community benefit or impact back to communities. Funding dissemination awards through the PACE mechanism directly supports and accelerates translational science by sharing results directly back to the community in meaningful ways.

OBJECTIVES/GOALS: Racial discrimination and its associated stress are well-documented contributors to health disparities among African Americans (AA).This feasibility study aimed to acquire methodological insights and build the infrastructure for a subsequent mindfulness interventional study to reduce the effects of racism-based stress. METHODS/STUDY POPULATION: 20 AA participants ( female 12, male 8 )ages 18-50 were enrolled, and clinical data (blood pressure, waist/hip, BMI, lipids, HbA1c, creatinine) for Allostatic Index were collected. Racism-based stress was measured using RBTSSS and the Everyday Discrimination Scale (EDS). Psychometric measures (Coping, resilience, mindfulness, social connection) and sleep (PSQI) were included. Bivariate analysis explored associations between psychological measures and stress biomarkers, supported by Spearman’s correlation analysis. RESULTS/ANTICIPATED RESULTS: Low discrimination (EDS) was associated with a lower Total cholesterol/HDL ratio (2.99 vs. 4.20, p=0.009) and higher HDL (62 vs. 52, p=0.001). Low EDS participants also had better sleep (mean=3, SD=1.33, vs. mean=5.8, SD=3.99, p=0.05*) but reported less coping through drugs and alcohol (p=0.022*) and higher resilience (p=0.047*). Mindfulness negatively correlated to sleep disturbance (r=-0.477 to r=-0.62), coping and resilience. . EDS correlated with overall life stress and drug and alcohol use. Sleep disturbance was negatively associated with social connection (r=-0.569**) and mindfulness. Sleep disturbance and discrimination correlated positively with drug and alcohol use and overall life stress (r=0.52 and r=0.0.59, respectively), while resilience was negatively correlated with sleep (r=-0.45). DISCUSSION/SIGNIFICANCE: Discrimination was associated with increased stress and unfavorable coping, while mindfulness may offer potential benefits for sleep, coping, and resilience. These preliminary findings provide a foundation for further exploring the potential of mindfulness interventions to address the biopsychological impacts of racism-induced stress.

OBJECTIVES/GOALS: To develop a loop-mediated isothermal amplification (LAMP) assay for the detection of cytomegalovirus (CMV) infection and drug resistance. METHODS/STUDY POPULATION: We designed core and loop primers sets utilizing the NEB LAMP Primer Design Tool. Each set contained four or six primers targeting major-immediate early genes – essential for viral entry/replication – or regions known to confer resistance to the antiviral drug ganciclovir. Optimization of reactions conditions was achieved employing DNA reference materials. Reactions were visualized through a change in color as amplification reactions accumulated. Successful reaction conditions were selected based on specific amplification products in less than 60 minutes. Limits of detection were evaluated as the main performance outcome. RESULTS/ANTICIPATED RESULTS: Genomic data were extracted and used to design a series of LAMP primers (48 total) that aimed to detect specific genomic regions of CMV. Using this strategy, we successfully designed and identified eight primer sets that showed high 100% sensitivity and 100 % specificity, when detecting > 1.00 x 10^5 copies/mL of CMV gDNA. We are in the process of characterizing a new set of primers to determine the diagnostic utility of a LAMP assay in detecting selected single-nucleotide mutations at the UL97 loci. The expected outcomes at completion include: (1) the identification of LAMP primers to detect drug-resistance mutants, (2) defining optimal conditions for successful reactions, and (3) determining limits of detection for subsequent validation with clinical specimens. DISCUSSION/SIGNIFICANCE: CMV infection remains one of the most dangerous infectious agents for immunocompromised hosts, newborns, and unborn children. This study will describe a proof-of-concept LAMP assay for the genotypic detection of drug resistance in CMV-infected individuals and hence, create new avenues for selection of effective therapies to treat CMV disease.

Objectives/Goals: The National Research Mentoring Network Coordination Center (NRMN CC) received funding from NIH to create an Online Community of Practice (OCoP) for Research Managers (RM). These RM contributed to advancing the research on the science of mentorship in STEMM. In a Proof of Concept, RM explored various aspects of their mentorship research support work. Methods/Study Population: An OCoP met 18 times across the 5-year grant to share experiences working to advance the science of mentorship in STEMM. Topics, frequency, and length of meetings were selected based upon several needs assessments surveys from the RM community. RM were invited to join the OCoP based on their roles as the point people for administrative activities in research projects (e.g., finances, budgets, training, and project management). RM often supervised staff, collected data, monitored IRB protocol compliance, and conducted research. RM played a pivotal role in the gathering and sharing of common measures across the 11 U01 studies to allow for greater confidence in research findings on mentorship (McConnell, 2021). Results/Anticipated Results: Our primary goal was to provide a supportive community for RM contributing to mentorship research and data sharing. The results from several needs assessments exemplified a request for this support from the community to engage in an OCoP focused on their professional development. Therefore, the OCoP served as a starting point to explore the duties, functions, roles and responsibilities of RM, and extended into providing professional development. Although the number of RM attending decreased as the grant entered the no-cost extension phase, a subgroup of RM expressed interest in continuing the OCoP to focus on dissemination of research findings, further supporting the need for this community. Discussion/Significance of Impact: RM supported 11 research projects and contributed to the data collection for over 6,000 participants. Few opportunities exist to create an OCoP for large scale data collection on experiences with mentorship in STEMM. RM were pivotal in their role with the NRMN to enhance the training and career development of individuals from diverse backgrounds.

OBJECTIVES/GOALS: Racial identity, one’s perception of that identity, and their perception of how others view their racial identity influences mental health. We aimed to assess the relationship between childhood trauma exposure, post-traumatic stress disorder (PTSD), and postpartum depression symptoms with individual Black identity in adulthood. METHODS/STUDY POPULATION: We examined whether racial identity, as measured by the regard subsection of the Multidimensional Inventory of Black Identity (MIBI), was impacted by childhood trauma exposure and related to PTSD and depressive symptoms in a sample of pregnant persons (N= 215, Mage=27.5, SDage=5.3) who identified within the Black diaspora, seeking prenatal care at Grady Hospital in Atlanta, GA. The regard subsection of the MIBI determines extent to which individuals feel positively about membership within the Black group (private regard) and the extent to which they think others feel positively about the Black group (public regard). We assessed childhood trauma using Childhood Trauma Questionnaire (CTQ) and depression and PTSD symptoms with Edinburgh Postnatal Depression Scale, and PTSD Checklist for DSM-5, respectively. RESULTS/ANTICIPATED RESULTS: Public regard (pubR) and private regard (privR) were significantly negatively correlated with childhood emotional abuse, emotional neglect, and overall childhood trauma exposure (all p’s.05). PrivR was significantly negatively correlated with both postpartum depression and PTSD symptoms (p DISCUSSION/SIGNIFICANCE: Ethnic-racial identity formation is a critical aspect of our psychological well-being, beginning in childhood and continuing across one’s lifespan. Our findings suggest that childhood trauma impacts racial identity in persons of the Black diaspora, which may contribute to negative mental health outcomes, including PTSD and depression.

OBJECTIVES/SPECIFIC AIMS: Leadership is an essential and recognized team science competency. To support the development of leadership skills at Mount Sinai, the LEAD (Leadership Emerging in Academic Departments) program, launched in 2016, delivers a structured 12-month blended learning program for junior faculty. The program aims to promote personal and professional leadership capacity, skills and behaviors. Following a competitive application process, 24 participants each year are selected to participate. In its second year, the challenge for the LEAD program leadership is to support alumni in fostering a culture of leadership that extends beyond the 12-month program. In order to promote a leadership community of practice and offer continued support to junior faculty, the LEAD Alumni program aims to bring former LEAD participants together to maintain motivation, share challenges and successes, meet with mentors and role models, and foster an ongoing community of practice that seeks to embed evidenced-based leadership culture at Mount Sinai. METHODS/STUDY POPULATION: The previous two cohorts of LEAD participants were approached to volunteer for the LEAD Alumni Forum working group. Four LEAD alumni came forward to form a self-selected working group. With input from the program leadership, the alumni working group is tasked with organizing regular events that bring the 48 previous LEAD participants together. The events provide the opportunity for individuals with expertise and a passion for leadership to create a supportive environment. This ultimately seeks to increase the transfer and utilization of leadership skills in practice, and promotes a culture of leadership. These alumni events also provide the opportunity for alumni to interact with senior leaders at Mount Sinai, thereby learning from role models within the organization. RESULTS/ANTICIPATED RESULTS: Evaluating learning transfer and culture change is challenging, so a number of proxy measures will provide insight into the success of the Alumni Forum. Firstly, the number of LEAD Capstone projects implemented in practice, and the success of these initiatives, will provide insight into transfer of leadership learning to practice. Secondly, participants will complete a validated survey tool, Leadership Programs Outcome Measure (LPOM), which explores self-reported leadership change at a personal, organizational and community level. Finally, participants will be followed up in the long-term to track promotion, awards, and other formal or informal leadership positions assumed following engagement in the LEAD program and the subsequent LEAD Alumni Forum. DISCUSSION/SIGNIFICANCE OF IMPACT: It is hoped the LEAD Alumni program will enhance the ability of participants to implement leadership knowledge and skills to practice, which may subsequently advance organization and culture change. Fostering a community of practice will further the reach of the LEAD program and as the number of LEAD alumni grows, and the Alumni Forum may provide the supportive environment that allows these individuals to have real impact.

ABSTRACT IMPACT: A comparative evaluation of the mechanical properties of commonly used posterior spinal fusion assemblies will allow surgeons to choose an assembly based on desired properties. The results will better inform surgical decision making and may lead to improved patient outcomes. OBJECTIVES/GOALS: The objective of this study is to evaluate and compare the mechanical properties of three posterior spinal fusion assemblies commonly used to cross the cervicothoracic junction. Fusion success depends on immobilization of vertebrae. The results will better inform surgical decision making and may improve patient outcomes. METHODS/STUDY POPULATION: Three titanium alloy posterior spinal fusion assemblies intended to cross the cervicothoracic junction underwent static compressive bending, tensile bending, and torsion as described in ASTM F1717 to a torque of 2.5 Nm: 3.5mm rods (Assembly A), 3.5mm to 5.5mm dilating rods (Assembly B), and two 3.5mm rods connected to two 5.5mm rods (Assembly C). Five samples of each assembly were attached to ultrahigh molecular weight polyethylene blocks via multiaxial screws for testing. The distance from the axis of rotation to the point of attachment of the rod and cervical screw was used as the lever arm to calculate the force required to create the desired torque for each test: lever arm of 37mm, requiring 67.6N of force to generate 2.5Nm of torque. Force and displacement were recorded, and stiffness of each construct calculated. RESULTS/ANTICIPATED RESULTS: An ANOVA was performed and resulted in p-values all p<0.005, indicating the test groups were significantly different. Therefore, pairwise t-tests with Bonferroni corrections (p<0.017) were used to determine pairs that were significantly different. Assembly A (3.5mm rods only) was found to be significantly less stiff than Assembly B (dilating rods) and Assembly C (3.5mm-connector-5.5mm rods) in each mode of bending: compression bending, tension bending, and torsion. Assembly A had a significantly greater range of motion in compression bending and torsion, but not tension bending, when compared to Assembly B and Assembly C. The only significant difference between Assembly B and Assembly C was found in the stiffness value of compression bending. DISCUSSION/SIGNIFICANCE OF FINDINGS: The results of this study indicate that incorporating a 5.5mm rod in a fusion assembly adds significant stiffness to the posterior spinal fusion construct. When stability of a fusion is of heightened concern, as demonstrated by the ASTM F1717 vertebectomy (worst case scenario) model, including 5.5mm rods increases the likelihood of fusion success.

OBJECTIVES/SPECIFIC AIMS: The goal of this project is to determine the role of ETV6 in early B-cell development and define how germline ETV6 mutations result in predisposition to leukemia. METHODS/STUDY POPULATION: Gene expression commons were queried for expression levels of Etv6 and Pax5 at different stages of hematopoiesis. Mouse bone marrow was isolated and fractioned into cells committed to the B cell lineage via B220+ and CD43+ staining by flow cytometry and then separated into the following fractions: Fraction A (CD24low, CD19−), Fraction B (CD19+, CD24+, BP1−), and Fraction C (CD19+ CD24+ BP1+). Wild-type or germline mutant P214L ETV6 were cloned in an MiG vector and expressed in Ba/F3 cells. ChIP-PCR was performed by cross-linking proteins to DNA with 1% formaldehyde for 10 minute at room temperature, followed by cell lysis with RIPA buffer. Lysates were sonicated to shear DNA to a length of 200–1000 base pairs, then Protein A agarose beads were used to clean and immunoprecipitate chromatin. RESULTS/ANTICIPATED RESULTS: We observed that Etv6 is highly expressed in hematopoietic stem and lymphoid progenitor cells through the pre-pro-B stage (FrA), but its expression is significantly reduced in fraction B and thereafter (p<0.0001). Etv6 expression decreases as B cells develop and is negatively correlated with Pax5 expression (r2=0.9993; p=0.0167). We next confirmed the expression patterns of ETV6 and PAX5 during B cell development in human samples. We found that ETV6 expression was higher in the early B cell fraction (CD10+, CD34+, CD19−, and CD20−) compared to the pre-B cell fraction (CD10+, CD34−, CD19+, CD20−). Conversely, we observed that PAX5 expression was higher in the preB cell fraction compared with the early B cell fraction. In Ba/F3 cells expressing ETV6 constructs, ETV6, but not ETV6 P214L overexpression significantly decreased Pax5 expression (p≤0.05). ETV6 is associated with the proximal GGAA site 72 base pairs upstream of the Pax5 TSS, but not GGAA sites further from the TSS. In addition, the transcriptional repressors SIN3A and HDAC3 were detected on the same regions of the Pax5 locus. We detected association of ETV6, SIN3A, and HDAC3 with the proximal GGAA site upon expression of WT ETV6, but not ETV6 P214L. DISCUSSION/SIGNIFICANCE OF IMPACT: Our results provide a mechanism of interaction for ETV6 and PAX5, 2 genes often disrupted in B-cell leukemia. These findings are significant because PAX5 misregulation results in a B cell development halt, lineage infidelity, and leukemogenesis. In continuing our studies, we have generated a transgenic mouse endogenously expressing the ETV6 P214L mutation by CRISPR/Cas9 editing, and these mice appear to have a thrombocytopenic phenotype similar to that observed in patients carrying the ETV6 P214L mutation. These animals will be the focus of our continued investigation of the mechanism by which ETV6 germline mutation results in a predisposition to leukemia. Our ultimate goal is a comprehensive understanding of how this process may be targeted more efficiently in patients with both heritable and sporadic forms of leukemia involving ETV6.