OBJECTIVES/GOALS: The goal was to develop foundational research knowledge and skills for clinicians interested in conducting clinical research. Emphasis was on the development of a research question and the iterative process necessary to transform a research question into a well-designed study and well-articulated research proposal for pilot grant funding. METHODS/STUDY POPULATION: The course took place over 10 sessions, May - September 2021. The application process required participants to provide an initial self-assessment of research skills and a proposed research question. 25 clinicians applied to the program, 11 were accepted, and 9 enrolled. All clinicians in the initial cohort were clinical faculty physicians. Because of the geographical distance of participants as well as the ongoing pandemic, the course used a blended learning approach with both synchronous and asynchronous learning. Participants viewed online lectures on core content coupled with live virtual sessions with opportunities for discussion and application of the content. Relevant CTSI and institutional resources were highlighted in each session. RESULTS/ANTICIPATED RESULTS: Of the 9 clinician participants who enrolled in Research 101, the average attendance per session was 67% or 6 people. 89% or 8 participants attended five or more sessions. 5 participants submitted a letter of intent for the annual CTSI Pilot grant program, and of the five, two were invited to submit a grant application. Formal evaluation of the program is currently ongoing and will close on November 30th, at which time full results will be available. Research competencies will be assessed through a pre-post comparison, each self-rated by course participants. Additionally, participants were asked to provide input on the most and least valuable components of the course, as well as any open-ended feedback. Research 101 leadership will use these results to improve the course for future participants. DISCUSSION/SIGNIFICANCE: A learning health system (LHS) is recognized as an essential means by which research evidence is translated into practice. Important to realizing the LHS vision is the engagement of clinicians into the generation and translation of research into practice. Research 101 is an important way to bolster clinician engagement in translational research.

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.

Ensuring appropriate review, approval, and oversight of research involving animals becomes increasingly complex when researchers collaborate across multiple sites. In these situations, it is important that the division of responsibilities is clear and that all involved parties share a common understanding. The National Institutes of Health Office of Laboratory Animal Welfare and the United States Department of Agriculture Animal Plant Health Inspection Service require an Institutional Animal Care and Use Committee (IACUC) to review the care and use of animals in research, and both agree that it is acceptable for one IACUC to review the work taking place at multiple institutions. With this in mind, several Harvard-affiliated hospitals and academic centers developed the Master Reciprocal Institutional Agreement for Animal Care and Use (Master IACUC Agreement) to support collaboration, decrease administrative burden, increase efficiencies, reduce duplicative efforts, and ensure appropriate protections for animals used in research. Locally, the Master IACUC Agreement has fostered greater collaboration and exchange while ensuring appropriate review and oversight of research involving animals. As multisite animal protocols become more prevalent, this Agreement could provide a model for a distributed, national network of IACUC reliance.

The 2020 COVID-19 pandemic has had a profound impact on the clinical research enterprises at the 60 Clinical and Translational Science Award (CTSA) Hubs throughout the nation. There was simultaneously a need to expand research to obtain crucial data about disease prognosis and therapy and enormous limitations on conducting research as localities and institutions limited travel and person-to-person contact. These imperatives resulted in major changes in the way research was conducted, including expediting Institutional Review Board review, shifting to remote interactions with participants, centralizing decision-making in prioritizing research protocols, establishing biobanks, adopting novel informatics platforms, and distributing study drugs in unconventional ways. National CTSA Steering Committee meetings provided an opportunity to share best practices and develop the idea of capturing the CTSA program experiences in a series of papers. Here we bring together the recommendations from those papers in a list of specific actions that research sites can take to strengthen operations and prepare for similar future public health emergencies. Most importantly, creative innovations developed in response to the COVID-19 pandemic deserve serious consideration for adoption as new standards, thus converting the painful trauma of the pandemic into “post-traumatic growth” that makes the clinical research enterprise stronger, more resilient, and more effective.

Translation of research discoveries into health impact can take many years, creating delays in improving clinical outcomes. One approach to promoting timely translation is to examine successful cases in order to understand facilitators and strategies for overcoming barriers. We examined the development of evidence-based management for neonatal abstinence syndrome (NAS) at one academic medical center, with a primary focus on pharmacologic treatment. Despite a substantial increase in NAS case incidence starting in the early 2000s, significant sociocultural, policy, and regulatory barriers limited collaborative NAS research. Facilitators for translation encompassed: 1) pursuing research of societal interest, 2) building an effective interdisciplinary team, 3) intentionally linking clinical, research, and advocacy efforts, 4) broad stakeholder engagement across clinical, policy, and research arenas, and 5) leveraging local resources. Challenges included lack of commercially available U.S. Food and Drug Administration approved neonatal drug formulations, legal and regulatory barriers related to off-label and illicit use of opioids, recruitment for a treatment associated with drug withdrawal syndromes, misalignment of research design needs with real-world scenarios, and episodic funding. Benefits of successful translation included improvements in clinical care, reduced healthcare costs related to NAS, and enhanced legislative, policy, and research strategies to support broader neonatal investigations.

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.

OBJECTIVES/GOALS: Sensorimotor integration deficits are common in Autism Spectrum Disorders (ASD). There is evidence for both an over-reliance on visual and proprioceptive feedback during motor control in ASD, suggesting deficits in the ability to modulate sensory feedback processing in order to use the most reliable input. This study aims to test this hypothesis. METHODS/STUDY POPULATION: 40 persons with ASD (ages 10-33 yrs) and 25 age-, sex- and nonverbal IQ-matched controls completed precision gripping tasks under multiple proprioceptive and visual feedback conditions. Participants squeezed a force sensor with their index finger and thumb and tried to match their force output to a target force. Visual feedback of the target force (stationary bar) and their force output (bar that moved up/down with increased/decreased force) were displayed on a computer screen. Visual feedback was presented across low, medium, and high gain levels; the force bar moved a greater distance per change in force at higher gains. Proprioceptive feedback was manipulated using 80Hz tendon vibration at the wrist to create an illusion that the muscle is contracted. Force regularity (approximate entropy; ApEn) was examined. RESULTS/ANTICIPATED RESULTS: We have scored data from 18 participants with ASD and 13 control participants to date. Preliminary results from these participants indicate a Group x Tendon Vibration x Visual Gain interaction for ApEn (F = 1.559, p = 0.023). Individuals with ASD show slight increases in ApEn with 80Hz tendon vibration relative to no tendon vibration in all visual conditions. Controls showed increased ApEn during 80Hz compared to no tendon vibration at low visual gain but decreased ApEn with tendon vibration at high visual gain. These preliminary results indicate that controls shift to using a secondary source of sensory feedback (e.g., proprioception) when the primary source (e.g., vision) is degraded. However, persons with ASD do not reweight different sensory feedback processes as feedback inputs are degraded or magnified. DISCUSSION/SIGNIFICANCE OF IMPACT: Our preliminary results reveal that sensorimotor issues in ASD result from deficits in the reweighting of sensory feedback. Namely, persons with ASD fail to dynamically recalibrate feedback processes across visual and proprioceptive systems when feedback conditions change. Our results may aid treatment development for sensorimotor issues in ASD.

OBJECTIVES/GOALS: Adolescence represents a critical period for substance use initiation. Various factors may contribute to trying a sip or single puff of a substance, that could lead to more frequent use. However, less is known about how predictors from multiple domains converge to impact risk for general substance use initiation. METHODS/STUDY POPULATION: The Adolescent Brain Cognitive Development (ABCD) study is a multi-site longitudinal study following youth into early adulthood. The present study included 7,644 ABCD children who reported no lifetime substance use (including any experimentation) at baseline (ages 9–10). Our primary aim was to use a random forest classification model to predict binary substance use initiation, defined as trying any non-prescribed substance (e.g., alcohol, tobacco, cannabis, non-prescribed medications), during a 2-year follow-up after baseline. A total of 402 variables from the following categories were examined as predictors: demographics, peer substance use and availability, mental and physical health, culture and environment, biospecimens, neurocognitive functioning, and structural neuroimaging variables. RESULTS/ANTICIPATED RESULTS: Over a two-year follow-up, 751 (9.8%) of substance-naïve children reported trying a substance by age 11. The most common substance was alcohol, followed by cannabis and tobacco. Mean Decrease Accuracy (MDA) values were used to assess the relative importance of each predictor. The overall accuracy of the model in accurately predicting group membership (no substance use initiation vs. substance use initiation) was 57.66%. Of the top 5 predictors, the most important predictor was intent to use alcohol (MDA = .002). The following top predictors were structural neuroimaging variables: volume and surface area of right lateral occipital lobe (MDA = .0009 and .0008, respectively), surface area of right inferior temporal lobe (MDA = .0007), and surface area of left superior frontal lobe (MDA = .0007). DISCUSSION/SIGNIFICANCE: A combination of intent to use alcohol and structural neuroimaging indices were among the top predictors of substance use initiation. Understanding predictors of early substance use experimentation is important for identifying at-risk youth that may require targeted intervention approaches.

OBJECTIVES/GOALS: Executive function (EF) deficits lead to poorer adherence and weight loss in obesity treatment. Conversely, untreated obesity leads to both EF impairments and risk of chronic disease. EF deficits in children with obesity have begun to be associated with biomarkers of chronic disease risk such as glucose and cortisol. Elevated pro-inflammatory biomarkers, such as C-Reactive Protein (CRP), Interleukin 6 (IL-6), and Tumor Necrosis Factor a (TNFa), indicate risk for obesity-associated chronic disease and may represent another candidate biomarker of EF deficits in pediatric obesity. These inflammatory markers tend to be elevated in obesity but have yet to be examined in association with EF in pediatric obesity. This represents an opportunity to identify potential biomarkers of EF which may serve as novel treatment targets that could improve outcomes and chronic disease prevention. Thus, the present study aims to explore associations between EF and blood serum CRP, IL-6, and TNFa. METHODS/STUDY POPULATION: Treatment-seeking children aged 8-12 years with BMI>95th percentile were recruited from a pediatric weight management clinic and attended baseline assessments for a family-based behavioral intervention program for pediatric obesity. Demographics and medical history were assessed via parent questionnaire. Height and weight were measured by research staff and converted to zBMI. Child performance-based EF was assessed via the NIH Toolbox Cognitive Battery iPad application. Blood draws, assays, and Dual-energy X-ray Absorptiometry (DXA; measuring adiposity) were conducted by trained research personnel. Pearson’s correlations were conducted to explore associations between EF (NIH Toolbox Cognitive Battery, fully corrected T-scores) and CRP, IL-6, and TNFa. RESULTS/ANTICIPATED RESULTS: Children (n = 12) were primarily female (76.5%) and African American (64.7%). Correlation coefficients between inflammatory markers and EF were highly variable (r = −0.39 to 0.52). CRP showed small, negative associations with Cognitive Flexibility (r = −0.39) and Inhibitory Control (r = −0.34). IL-6 also demonstrated a small, negative association with Inhibitory Control (r = −0.37). TNFa was positively and moderately associated with working memory (r = 0.53). Remaining associations were weak (−0.3<r<0.3). DISCUSSION/SIGNIFICANCE OF IMPACT: Signals of higher inflammation, measured via CRP and IL-6, in association with EF deficits were identified in a small sample of children with obesity, as hypothesized. However, signals in the opposite direction were identified as well, measured via TNFa. CRP and IL-6 may represent candidate biomarkers of executive dysfunction in obesity that warrant further domain and biomarker-specific research, with potential long-term implications for improving pediatric obesity treatment.

OBJECTIVES/GOALS: The Workforce Development Core of the New Jersey Alliance for Clinical and Translational Science (NJ ACTS) has developed an internship program for students to engage in virtual research projects across the CTSA Hub. We sought to evaluate how intern projects within NJ ACTS align with the competencies needed for translational science. METHODS/STUDY POPULATION: Core leads and researchers within NJ ACTS developed 34 projects that were completed by individual interns or pairs of interns. Forty-two professional, undergraduate, and graduate students across the 3 Hub institutions have completed semester-long intern projects. Intern mentors mapped their projects to the C-COMEND competency profile for translational scientists which were further aligned with the seven fundamental character traits defined by Translation Together. RESULTS/ANTICIPATED RESULTS: More than 75% of intern projects addressed the C-COMEND competencies in Personal Development, Communication and Dissemination, Project Management and Time Management. Few projects (< 10%) focused on skills related to preclinical or clinical research. The competencies needed for development as a Rigorous Researcher were most consistently addressed in the intern projects. Additionally, intern projects fostered a number of skills needed for becoming a Domain Expert and Skilled Communicator. DISCUSSION/SIGNIFICANCE: Taken together, a virtual internship program can be designed to introduce and/or refine the competency skills needed for translational science.

OBJECTIVES/GOALS: The objective of this evaluation is to show how the STEP.UP program promoted the professional development at Michigan Medicine by providing clinical and translational research staff an experienced research staff mentor in a structured 9-month program. METHODS/STUDY POPULATION: Participant and mentor data was collected from application forms, online surveys, and interviews with both participating mentors and mentees. Validated assessments of mentoring competencies were administered. Participants were tracked over a period of four years with regular reviews of institutional records. Mentor and mentor data was also collected at the point of application each year and the application forms were aligned with NIH definitions for underrepresented populations in science in 2020. As part of a process of continuous programmatic improvement, a STEP.UP Advisory Board consisting of senior research staff and past mentors was involved in the identification, operationalization and evaluation of programmatic outcomes and is involve din the ongoing governance of this mentoring program. RESULTS/ANTICIPATED RESULTS: Four cohorts of mentees and mentors have participated in this program since its inception. Mentees gained the greatest abilities in, Active listening, Establishing a relationship based on trust, Considering how personal and professional differences may impact expectations, and Working effectively with mentors/mentees whose personal background is different. Mentees reported the program contributed to their Career planning, Professional advancement, networking, personal growth, professional networks, and communication skills. Mentors reported learning about new professional techniques and areas of expertise. As of 2021, 75% the first cohort changed their job-classification since participating as did 25% of the second cohort and 100% of mentees have maintained research careers. DISCUSSION/SIGNIFICANCE: The creation of this program in 2019 marked the beginning of a novel professional development opportunity at Michigan Medicine. The evaluation results show how STEP.UP contributes to advancing clinical and translational study teams and how it can inform and the identification of best practices in clinical and translational workforce development.

OBJECTIVES/GOALS: Fluid boluses are administered to hypotensive, critically ill children but may not reverse hypotension, leading to delay of vasoactive infusion, end-organ damage, and mortality. We hypothesize that a machine learning-based model will predict which children will have sustained response to fluid bolus. METHODS/STUDY POPULATION: We will conduct a single-center retrospective observational cohort study of hypotensive critically ill children who received intravenous isotonic fluid of at least 10 ml/kg within 72 hours of pediatric intensive care unit admission between 2013 and 2023. We will extract physiologic variables from stored bedside monitors data and clinical variables from the EHR. Fluid responsive (FR) will be defined as a MAP increase by 310%. We will construct elastic net, random forest, and a long short-term memory models to predict FR. We will compare complicated course (multiple organ dysfunction on day 7 or death by day 28) between: 1) FRs and non-FRs, 2) predicted FRs and non-FRs, 3), FRs and non-FRs stratified by race/ethnicity, and 4) FRs and non-FRs stratified by sex as a biologic variable. RESULTS/ANTICIPATED RESULTS: We anticipate approximately 800 critically ill children will receive 2,000 intravenous isotonic fluid boluses, with a 60% rate of FR. We anticipate being able to complete all three models. We hypothesize that the model with the best performance will be the long short-term memory model and the easiest to interpret will be the tree-based random forest model. We hypothesize non-FRs will have a higher complicated course than FRs and that predicted non-FRs will have a higher rate of complicated course than FRs. Based on previous adult studies, we hypothesize that there will be a higher rate of complicated course in patients of black race and/or Hispanic ethnicity when compared to non-Hispanic white patients. We also hypothesize that there will be no difference in complicated course when comparing sex as a biologic variable. DISCUSSION/SIGNIFICANCE: We have a critical need for easily-deployed, real-time prediction of fluid response to personalize and improve resuscitation for children in shock. We anticipate the clinical application of such a model will decrease time with hypotension for critically ill children, leading to decreased morbidity and mortality.

OBJECTIVES/GOALS: Diagnostic odyssey is the time it can take to a patient for receiving a diagnosis. Diagnostic process in rare diseases can be complex due to the heterogeneity of symptoms and lack of access to care. We aim to evaluate the association between diagnostic odyssey, perceived stress, and access to care, in parents of Puerto Rican patients with a rare disease. METHODS/STUDY POPULATION: We propose a cross-sectional study in parents of 100 children who received an uninformative whole exome sequencing (WES) report during a scheduled appointment with their geneticist. Discussion of WES results during clinical session, followed by a Perceived Stress Scale (PSS-10) and semi-structured interview to explore the experience of access to care during the diagnostic process will be arranged. Observation and interviews will be recorded. Data analysis and descriptive statistics will be calculated using STATA. Statistical associations (OR) will be estimated using generalized linear models at a 5% significance level. RESULTS/ANTICIPATED RESULTS: We expect to find high perceived stress in parents of Puerto Rican pediatric individuals having rare diseases, especially among single mothers. We will be able to identify limited access to care in Puerto Rico, especially in the testing pre-authorization process and long waits for geneticist appointments. Demand for advanced diagnostics is above the number of medical geneticists available in Puerto Rico, which triggers delayed diagnosis, management, and counseling. Therefore, these could affect the health disparities in our population with rare diseases. DISCUSSION/SIGNIFICANCE: This descriptive study will evaluate perceived stress in parents of pediatric patients living a diagnostic odyssey in Puerto Rico. No study has described perceived stress and access to care in this Hispanic population with undiagnosed conditions. Findings will contribute to a deep understanding of diagnostic process and limited access to care.

ABSTRACT IMPACT: This project aims to better understand mechanisms of sensory and motor deficits in individuals with ASD with the goal of informing diagnosis and treatment development. OBJECTIVES/GOALS: Over-reliance on both visual and proprioceptive feedback have both been observed during motor behavior in persons with Autism Spectrum Disorders (ASD), suggesting that separate sensory feedback processes may be selectively altered during different behaviors. The objective of this study is to clarify sensory mechanisms of fine motor control in ASD. METHODS/STUDY POPULATION: Participants with ASD (N=43) and controls (N=23) matched on age (10-20 yrs) and non-verbal IQ completed tests of precision gripping. Participants were instructed to press on force sensors with their index finger and thumb so that a moving bar corresponding to their force output reached and stayed as stable as possible at the level of a stationary target bar. Visual feedback was manipulated by changing the visual gain of the force bar (low, medium and high). The force bar moved more per change in force output at higher gains. Proprioceptive feedback was manipulated by applying 80 Hz tendon vibration at the wrist to induce an illusion of muscle contraction. This was compared to a condition with the tendon vibrator turned off. Force variability (standard deviation) and regularity (sample entropy) were examined. RESULTS/ANTICIPATED RESULTS: Controls showed increased force variability with the tendon vibration on compared to off (t = -3.372, p < 0.001); however, the ASD group showed no difference in force variability between the tendon vibration conditions (t = -0.960, p = 0.338). Individuals with ASD had stronger age-associated reductions in force variability relative to controls across tendon vibrator and gain conditions (Group x Age: t = -4.05, p < .001). The ASD group also had greater age-associated increases in force regularity relative to controls, especially at higher gain levels (Group x Gain Level x Age: t = -3.22, p = 0.001). Unlike the ASD group for whom regularity increased with age in both tendon vibration conditions, controls only showed these age-related gains when the tendon vibrator was off (Group x Vibration Frequency x Age: t = 2.46, p = .014). DISCUSSION/SIGNIFICANCE OF FINDINGS: Our findings indicate that while controls integrate proprioceptive and visual feedback online to accurately adjust fine motor behavior, persons with ASD rely mostly on visual feedback. Our results suggest delayed development of sensory integration and reduced reliance on multisensory feedback during online fine motor control in persons with ASD.

OBJECTIVES/GOALS: The management of head and neck cancers is complicated and allows for a variety of disparate approaches from providers. However, data from several hundred or thousands of patients is necessary to decipher the optimal decisions for populations of patients. And prospective trials would take years to accrue and often are financially not possible. METHODS/STUDY POPULATION: Instead, we collated the entire electronic medical record for all patients at our institution treated for head and neck cancers. We employed a variety of clinical informatics and natural language processing to gather text data into large data frames. We found key conclusions in the diagnostic, treatment, and surveillance of our patients. RESULTS/ANTICIPATED RESULTS: First, obtaining post-operative PET/CT changes in management in over one-third of patients, highlighting the utility of optimizing diagnostic imaging. Second, using a newer silicone-based cream instead of just a moisturizer decreased the absolute risk of grade 2+ radiation dermatitis by almost 15%. Lastly, we are deploying novel autosegmentation frameworks to better understand tissue decomposition in the head and neck to identify patients in need of further nutritional support while undergoing radiation therapy. DISCUSSION/SIGNIFICANCE: Collectively, we showcase the value and opportunity of mining oncological data for the improvement of patient care.

OBJECTIVES/SPECIFIC AIMS: The study aims to identify patient and provider factors associated with delay in diagnostic resolution after an abnormal screening mammogram, with an emphasis on whether patients who spoke Chinese as their primary language sustained longer times to resolution. Primary outcome is to identify what proportion of patients achieve diagnostic resolution after abnormal screening mammogram within 90 days. Secondary outcome is to identify whether Chinese-speaking patients experience longer times to diagnostic resolution. METHODS/STUDY POPULATION: We performed a single-center retrospective cohort study at Tufts Medical Center (TMC), a tertiary care hospital that serves as the primary referral site for the Chinatown neighborhood in Boston. We included patients who underwent screening mammogram between 10/1/2015-9/30/2016 which was resulted as BIRADS-0 (non-diagnostic). Diagnostic resolution was defined as BIRADS-1, 2, or 3 imaging or definitive biopsy. We collected data on patient demographics (age, insurance plan, race/ethnicity, primary language, history of cancer), provider characteristics (referring provider location), and post-referral testing. Insurance was categorized as private-only or subsidized. Poverty was categorized using the American Fact Finder database, with a binary variable of <20% of ≥20% people in poverty for a given zip code. We performed descriptive statistics for all variables. We will perform multivariable Cox regression analyses to determine whether Chinese-speaking patients experience longer time to diagnostic resolution, adjusting for age, referring provider type, insurance status, poverty, and breast cancer history. We will use p<0.05 for our threshold for significance. RESULTS/ANTICIPATED RESULTS: We identified 386 patients who met inclusion criteria. Over half (55.9%) of patients were Caucasian, the mean age of study patients was 59 years, and 22% of patients were classified as poor. English was the most commonly spoken primary language (77.7%), while 15.3% of patients identified a Chinese dialect as their primary language. Most patients solely used private insurance for their medical care (73.1%). Majority of patients (83%) presented after undergoing a routine screening mammography, but a considerable proportion (14.4%) had prior breast cancer or a palpable mass. Most patients were referred for their screening mammogram by a hospital-based provider at TMC (85%), of which 77% of TMC referrals were from primary care. We also noted a limited number of referrals from community health centers, private practices and other PCP’s (Table 1). We will calculate median time to diagnostic resolution after screening mammogram and the proportion of patients who achieve resolution within 90 days. We will also calculate time to initiation of diagnostic workup, and whether this differed among Chinese-speaking patients, subsidized patients, or among those who were referred from outside of TMC. We will complete Cox multivariable analysis to identify if Chinese-speaking patients experience longer time to diagnostic resolution, adjusting for age, insurance status, Primary care provider location, poverty, and prior history of breast cancer. We will a priori test for an interaction between primary care provider within Tufts and Chinese as primary language to identify if a PCP within TMC modifies the relationship between Chinese language and time to resolution. DISCUSSION/SIGNIFICANCE OF IMPACT: The proposed study will identify whether disparities exist in time to achieving diagnostic resolution. Specifically, we will identify if patients who are primarily Chinese-speaking experience longer time to resolution. Our results will potentially provide the foundation for a patient navigation program to attenuate existing disparities by providing additional support for Chinese speaking patients in breast imaging workup.

OBJECTIVES/GOALS: Vitamin D [25(OH)D], known to have anti-inflammatory and anti-fibrotic effects in other tissues, may also impact adipose tissue. We designed parallel studies in humans and rodents to define the effects of vitamin D on adipose tissue inflammation and fibrosis, and on systemic insulin resistance. METHODS/STUDY POPULATION: We performed a randomized, double-blinded placebo-controlled trial to examine the effects of repleting vitamin D at to two levels (to >30 ng/ml and to > 50 ng/ml) in 25(OH)D-deficient (<20 ng/ml), insulin resistant, overweight-to-obese humans (n = 19). A comprehensive study of whole-body insulin action was undertaken with euglycemic stepped hyperinsulinemic clamp studies, both before (1st visit) and after administration of vitamin D or placebo (2nd visit and 3rd visit). Adipose tissue fibrosis and inflammation were quantified by ‘real-time’ rt-PCR and immunofluorescence. To determine whether vitamin D’s effects are mediated through adipocytes, we performed hyperinsulinemic clamp studies and adipose tissue analysis in an adipocyte-specific vitamin D receptor knockout (VDR KO) mouse model. RESULTS/ANTICIPATED RESULTS: 25(OH)D repletion (to >30 ng/ml) was associated with reductions in adipose tissue expression of inflammatory (0.6-0.7-fold decreased expression of TNF-α, IL-6, iNOS and PAI-1) and pro-fibrotic (0.4-0.8-fold decreased expression of TGF-β1, HiF1α, Collagen I, V, VI and MMP7) factors, decreased collagen VI immunofluorescence (p = 0.02) and improved hepatic insulin sensitivity in humans, with suppression of endogenous glucose production (EGP) (1.28 ± 0.20 vs 0.88 ± 0.18 mg/kg/min, p = 0.03). Compared to wild type (WT), VDR KO mice exhibited increased adipose tissue expression of several pro-inflammatory (Tnf-α, iNos, Pai-1, Mcp-1 and F4/80; 4-10 fold) and pro-fibrotic genes (Tgf-β1, Collagen VI, and Tsp1; 2-4 fold), in concert with hepatic insulin resistance (EGP 10 ± 3 vs 3 ± 2 mg/kg/min in WT, p = 0.021). DISCUSSION/SIGNIFICANCE OF IMPACT: Collectively, these complementary human and rodent studies establish a beneficial role of vitamin D to improve hepatic insulin resistance, likely by restraining adipose tissue inflammation and fibrosis. Thus, normalizing 25(OH)D levels could have metabolic benefits in targeted individuals. CONFLICT OF INTEREST DESCRIPTION: N/A

OBJECTIVES/GOALS: Many ophthalmologic procedures involve operating on or manipulating the globe and bony orbit. Creating an anatomically accurate globe and orbit is of interest to improve surgical education for trainees. The purpose of this study was to create a high-fidelity globe and orbit model using synthetic materials and utilizing 3D-printing techniques. METHODS/STUDY POPULATION: A deidentified computed tomography scan of the head and neck was digitally rendered and segmented using Mimics and 3-Matic (Materialise NV, Belgium) to create a digital model of the bony orbit. The model was 3D printed using a stereolithographic 3D-printer (Formlabs, Somerville, MA). The globe was created by soaking a large water bead made from a water absorbing polymer (YIQUDUO, China) for 24 hours. The water bead was then coated consecutively with three layers of silicone (Smooth-On, Macungie, PA). A standard sausage casing was hydrated and encased around the water bead, representing the conjunctiva. The globe was placed into the 3D-printed orbit. An incision was made in the sausage casing and the defect was sutured by one ophthalmologist. RESULTS/ANTICIPATED RESULTS: The bony orbital anatomy was accurately represented by stereolithographic printing. The size and feel of the artificial globe was similar to that of an in vivo human globe. The incised sausage casing covering the globe was able to be manipulated and sutured using a 8-0 suture in a microsurgical environment. The sausage casing had high-fidelity characteristics of an in vivo human eye conjunctiva. DISCUSSION/SIGNIFICANCE: This model can be used for teaching of conjunctival suturing for ophthalmologic trainees. By use of easily obtained materials for the globe, this model has the potential to standardize teaching methods of challenging techniques, and can reduce the need for animal and human tissue procurement, which is the current standard for ophthalmologic teaching.

ABSTRACT IMPACT: Knowledge of which aspects of social connectedness most strongly associate with caregiver health and health behaviors can inform intervention targets to improve caregiver health OBJECTIVES/GOALS: Stressed dementia caregivers are at risk of poor health. Social connectedness may reduce adverse health effects, yet it is unknown about which aspects relate most strongly to health. This is a barrier to intervention development. Our study identifies aspects of social connectedness most strongly associate with caregivers’ daily health behaviors. METHODS/STUDY POPULATION: Data. Enrolled spousal caregivers completed 14 consecutive days of online surveys. Measures. We examined multiple health behaviors each day, which included: 1) number of occurrences of 3 potential binge-eating behaviors (range 0 to 30), 2) whether participants engaged in at least 30 minutes of physical activity, and 3) perceived sleep quality, rated 1 (very bad) to 5 (very good). We also examined a count of health symptoms caregivers experienced (e.g., backache; range: 0 to 7). Measures of social connectedness included: spousal emotional support, perceived spousal appreciation, emotional support from any source, and loneliness. Analysis. We applied bivariate multi-level mixed effects models to examine the association between each aspect of social connectedness and health behaviors day-to-day. RESULTS/ANTICIPATED RESULTS: Since November 2020, 5 of N=40 participants were enrolled, of whom 3 had completed all diary surveys. Participants were women ages 59 to 73, and included 4 non-Hispanic white and 1 Hispanic caregivers. Data included 51 days of surveys (93% adherence). No differences in eating behaviors nor physical activity according to social connectedness were found. Emotional support from any source was positively associated with sleep quality (B= 0.20; SE=0.08; p-value 0.015). On days when caregivers indicated their spouse appreciated them ‘A lot’ versus ‘Not at all,’ sleep quality was marginally better (B=0.73, SE=0.42; p-value-0.08). Finally, days when caregivers felt lonely at least ‘Some of the time’ versus ‘Not at all’ were associated with experiencing more adverse health symptoms (B=1.54; SE=0.58; p-value<0.001). DISCUSSION/SIGNIFICANCE OF FINDINGS: Improved emotional support from any source may support better sleep quality among caregiving spouses, while loneliness appears to contribute to more adverse health symptoms. Findings, if confirmed, can be translated to develop intervention programs that target loneliness and perceived emotional support among caregivers.