OBJECTIVES/SPECIFIC AIMS: Congenital heart disease (CHD) is the most frequently occurring birth defect in the United States affecting about 40,000 infants born every year. Despite significant advances in postsurgical survival, developmental outcomes remain disproportionately poor. Therapeutic hypothermia has been used for neuroprotection during cardiac surgery since the 1950s. Infants undergoing cardiac surgery are typically cooled to 28–33°C during the operation and then rapidly rewarmed to normothermia following surgery at a rate of 1°C every 3–5 minutes to minimize concerns surrounding the risks associated with prolonged bypass exposure. However, emerging evidence from animal models has shown rapid temperature changes following surgery may diminish or even negate the neuroprotective effect of intraoperative hypothermia. No prospective studies have assessed the safety or impact of alternative approaches to postoperative temperature management on the outcome of infants with CHD undergoing cardiac surgery. Therefore, we conducted a pilot study to examine the safety of a novel application of a temperature-regulating device to slowly rewarm infants with congenital heart disease over the 12 hours following cardiac surgery. METHODS/STUDY POPULATION: From November 2014 to July 2016, infants with CHD requiring surgery with cardiopulmonary bypass before the age of 12 months were prospectively recruited. Infants were randomized in blocks of 3 with 1 allocated to standard of care and 2 to the experimental protocol. Infants assigned to the standard of care were rewarmed in the operating room while on bypass at a rate of 1°C every 3–5 minutes back to a temperature of 37°C. Infants assigned to the experimental intervention, were rewarmed on bypass to 35°C and then over the subsequent 12 hours following surgery, gradually rewarmed using an FDA approved “cooling blanket” to increase temperature by 0.3°C every 2 hours for 6 hours and then by 0.2°C every 2 hours for 6 hours until the goal temperature of 36.5°C was achieved. Frequency of serious, moderate and other adverse events were tracked. Detailed vital sign data was collected hourly for the first 12 hours after surgery and then every 6 hours for the next 36 hours and included temperature, highest and lowest heart rate, highest and lowest systolic blood pressure, and highest and lowest diastolic blood pressure. Presence or absence of abnormal cardiac rhythms was recorded per 24-hour interval. Chest tube output was recorded in cc/kg/8 hours for as long as the chest tube was in place. Laboratory data points included serum creatinine level, serum glucose level, liver function tests (AST and ALT), platelet count, hematocrit level, PTT, INR, fibrinogen, white blood cell count and lactate. Blood samples for biomarkers of brain injury (s100b and NSE) were obtained on all infants at the following 4 intervals; the preoperative setting for baseline, postoperatively after bypass, on postoperative day 1, and on postoperative day 2. For this safety study, the primary outcome measure was a composite outcome of the frequency of serious adverse events as well as the frequency of any adverse events and was compared among treatment groups. Data were analyzed using an intent to treat analysis. The study was approved by the Maine Medical Center Institutional Review Board. RESULTS/ANTICIPATED RESULTS: Seven infants were randomized to the standard of care group and 9 were randomized to the experimental group. There were 2 exclusions after randomization in the standard of care group with 1 death in the operating room and 1 unsuccessful attempt to wean from bypass. The mean temperature upon arrival to the PICU for the experimental infants was 35.2°C (range 34–36°C) and for the standard of care infants was 37.5°C (range 36.9–38.9°C). For the first 8 hours after surgery, infants in the standard of care group had mean temperatures over 37.0°C. There were no significant differences in the frequency of serious, moderate, or other adverse events between the standard of care group and experimental group. No infant in either group had need for cardiopulmonary resuscitation or exploratory surgery within 48 hours following surgery nor did any infant experience any clinically appreciated adverse neurological events such as stroke or seizure. No infant in either group experienced clinically significant bradycardia of less than 100 beats per minute or sustained tachycardia of greater than 160 beats per minute. There was a trend toward lower heart rates in the experimental group. Junctional Ectopic tachycardia (JET) occurred in 2 patients in the experimental group and 1 in the standard of care group. The mean highest INR in both groups was 1.4 (range 1.2–1.6). The mean lowest recorded platelet level in the first 48 hours was 128.8 (range 87–160) in the standard of care group and 123.8 (range 49–229) in the experimental group. Infants in the experimental group had lower chest tube output overall than the standard of care infants. The mean days of intubation for standard of care infants was 5 days (range 1–15 days) and for experimental infants the mean was 3.7 days (range 0–16 d). The PICU length of stay was shorter for the experimental infants (6.9 vs. 12 d for standard of care). The total length of stay was also shorter for experimental infants (12.4 vs. 16.4 d for standard of care). Serum biomarkers of brain injury (s100b and Neuron specific enolase) were elevated in the immediate postoperative period for infants in the standard of care group compared with the experimental group but normalized more quickly for standard of care. DISCUSSION/SIGNIFICANCE OF IMPACT: This small pilot study suggests that mild hypothermia following congenital heart surgery in infants under the age of 12 months is safe as there was no increase in the rate of severe, moderate, or other adverse outcomes in infants who received the experimental treatment of delayed rewarming. This study provides evidence for the efficacy of the cooling blanket in regulating the temperature of infants after surgery. Trends toward lower chest tube output, shorter intubation and decreased length of stay are possibly the result of improved hemodynamic stability in the absence of postoperative fever. Future studies will need to assess the effect of mild hypothermia compared with a normothemic control group.

ABSTRACT IMPACT: This study helps translate investigational bedside optical coherence tomography into improved diagnosis and care of preterm infants at risk for retinopathy of prematurity. OBJECTIVES/GOALS: In retinopathy of prematurity screening, fundus photography may be of limited quality in eyes with dark fundus pigmentation (FP). The goal of this study was to evaluate the impact of FP on overall scan quality and retinal layer visibility on investigational bedside optical coherence tomography (OCT) in preterm infants. METHODS/STUDY POPULATION: We analyzed 846 OCT scans captured prospectively from 188 eyes of 94 preterm infants enrolled in the BabySTEPS study (NCT02887157). Trained ophthalmologists, masked to OCT findings, determined FP (blond, medium, or dark). Expert graders, masked to FP, evaluated OCT images for: 1) overall OCT quality (excellent, acceptable, poor, or unusable); and 2) all age-appropriate retinal layers visible (yes or no). To assess the association of FP with OCT quality (excellent/acceptable or poor/unusable) and retinal layer visibility, we performed multivariable logistic regression modeling, adjusting for biologic and demographic confounders and correlations from repeated OCT scans and paired eyes. RESULTS/ANTICIPATED RESULTS: Mean birthweight was 964 (SD=283) grams, mean gestational age was 27.8 (SD=2.6) weeks, 48 (51%) infants were male, and 51 (54%) were non-white. On exam, 72 (38%) eyes had blond FP, 92 (49%) had medium, and 24 (13%) had dark. OCT quality was excellent or acceptable in 725 scans (86%) and all age-appropriate retinal layers were visible in 781 scans (92%). Compared to eyes with blond FP, eyes with medium and dark FP did not have higher odds of poor/unusable OCT scan quality (adjusted OR 0.87 [95% CI 0.50-1.48] and 0.49 [95% CI 0.16-1.55], respectively) or not all age-appropriate retinal layers visible on OCT (adjusted OR 1.17 [95% CI 0.39-3.51] and 0.57 [95% CI 0.15-2.20], respectively). DISCUSSION/SIGNIFICANCE OF FINDINGS: Medium and dark FP did not impact overall scan quality or age-appropriate retinal layer visibility on investigational bedside OCT in preterm infants. This study supports the feasibility of using OCT to analyze retinal microanatomy in diverse populations of preterm infants with a range of FP.

OBJECTIVES/SPECIFIC AIMS: The purpose of this study was to provide individuals who have experience with cognitive behavioral therapy (CBT) with a demonstration of how using their therapeutic strategies affects their brain activity. Two challenges that face CBT and other cognitive therapies are (1) sustaining the gradual, incremental behavioral changes characteristic of the treatment and (2) measuring associated biological changes. These challenges may impede treatment efficacy and may negatively affect treatment outcomes, including patient discontinuation of CBT. Ideas for addressing these issues include providing patients with (1) a more immediate indicator of therapy effectiveness as well as (2) a biological index of behavioral change. In this study, we aimed to provide participants with an index of biological change based on therapeutic experiences via use of real-time functional magnetic resonance imaging (rtfMRI) neurofeedback. METHODS/STUDY POPULATION: We recruited participants who had already completed cognitive therapy as part of a clinical trial for depression at the University of North Carolina at Greensboro (n=13). In the present experiment, participants were asked to provide a list of negative autobiographical memories or worries as well as cognitive strategies they use to cope with negative moods. The task consisted of COUNT, MEMORY, and STRATEGY trials (30 s each). During baseline COUNT trials, participants counted backwards (e.g., 300–4). During MEMORY trials, they viewed phrases previously developed describing their negative autobiographical memories/worries. During STRATEGY trials participants viewed a strategy they use to help them process the memory/worry. First, a localizer run was completed to determine a unique region of interest for each participant. We identified peak activation within the cingulate cortex to the contrast of MEMORY (STRATEGY+COUNT). Although the task was the same, no neurofeedback was displayed during the localizer run. During the feedback runs, participants were shown neurofeedback from the cingulate cortex following both the MEMORY and STRATEGY trials. This activation was represented on a signal bar display and represented the average cingulate activation during the trial. Unlike many rtfMRI studies, the purpose here was not for participants to interact with the neurofeedback directly. Rather, a feedback summary was shown to participants after each MEMORY and STRATEGY trial as an index of how brain activity changed in response to negative memories/worries and therapeutic strategies. Our goal was not for participants to learn to self-regulate the cingulate cortex, but rather to provide participants with a metacognitive demonstration of strategy efficacy. Participants were given detailed instructions regarding the task design, the role of the cingulate cortex in depression, as well as the hypothesized direction of activation during the MEMORY and STRATEGY phases to help them interpret the neurofeedback. RESULTS/ANTICIPATED RESULTS: Results revealed that “stronger neurofeedback” (defined as the difference between STRATEGY vs. MEMORY trials) correlated with self-reported strategy efficacy ratings immediately following the scan session (p<0.05). More importantly, stronger neurofeedback predicted both self-reported strategy efficacy and frequency of use 1 month following the MRI session (p<0.05). Importantly, this relationship was specific to only those strategies used inside the scanner; and no such relationship was observed at baseline. Neuroimaging results revealed that during the MEMORY phase, activation within inferior frontal gyrus and supramarginal gyrus correlated with baseline BDI score (whole brain, cluster corrected with FSL Flame 1 to p<0.05). During the STRATEGY phase, the periaqueductal gray nucleus, insula, and temporal pole predicted self-reported frequency of strategy use 1 month post-scan session (whole brain, cluster corrected with FSL Flame 1 to p<0.05). DISCUSSION/SIGNIFICANCE OF IMPACT: We believe this study holds promise to provide a powerful demonstration for individuals that strategies used to cope with negative moods can produce significant changes in their brain.

OBJECTIVES/GOALS: Previous research has shown acute and chronic alcohol effects on cardiac function, including elevated heart rate (HR) and lowered heart rate variability (HRV). This study aimed to examine the relationship between cardiac reactivity and subjective response following intravenous (IV) alcohol in non-dependent drinkers. METHODS/STUDY POPULATION: Non-dependent drinkers (N = 46, average age = 25.2) completed a human laboratory IV alcohol self-administration (IV-ASA) session. Subjective response to alcohol was assessed using the Drug Effects Questionnaire (DEQ) and Alcohol Urge Questionnaire (AUQ). Drinking behavior was assessed using the Alcohol Timeline Followback (TLFB) and Alcohol Use Disorders Identification Test (AUDIT). HR was recorded using the Polar Pro Heart Rate monitor throughout the session. HRV measures were calculated using guidelines determined by the Task Force of the European Society of Cardiology and The North American Society of Pacing and Electrophysiology. RESULTS/ANTICIPATED RESULTS: Recent drinking history as measured by the AUDIT and TLFB was not significantly different by sex. Results showed heavier drinking measures (AUDIT and TLFB) were positively associated with HRV measures (all p-values < 0.02). Those who reported a greater increase in alcohol craving (AUQ score) and wanted more alcohol (DEQ) following an alcohol prime, showed a greater change in HRV (p < 0.005). When examining HRV change from baseline throughout the priming session, there was a significant sex interaction for NN50 (p < 0.03) and a trend for PNN50 (p-value < 0.07). DISCUSSION/SIGNIFICANCE OF IMPACT: Acute IV alcohol alters cardiac reactivity measures in non-dependent drinkers. Future directions include examining the role of sex in HRV changes during alcohol consumption during IV-ASA. Understanding the effect of alcohol on cardiac reactivity and physiology may help characterize those at risk for alcohol use disorders.

OBJECTIVES/SPECIFIC AIMS: There are currently no established blood-based biomarkers of recovery and neural repair following stroke in humans. Such biomarkers would be extremely valuable for aiding in stroke prognosis, timing rehabilitation therapies, and designing drugs to augment natural repair mechanisms. Metabolites, including lipids and amino acids, are engaged in many cellular processes and cross the blood-brain barrier more easily than proteins. Recent advances in liquid chromatography / mass spectrometry (LCMS) allow researchers to obtain a biochemical fingerprint of the metabolites in various biofluids. Thus, metabolite biomarkers of neural repair after brain injury are a promising avenue for future research. Objective: Design and conduct a study to identify metabolite changes in the blood associated with good and poor motor recovery following stroke. METHODS/STUDY POPULATION: We launched the Biomarkers of Stroke Recovery (BIOREC) study, which seeks to enroll 70 participants suffering arm motor impairment following stroke and 35 matched controls. BIOREC is a longitudinal observational study. Fasting blood samples are collected at 5, 15, and 30 days post-stroke, processed, and stored in the Georgetown Lombardi biorepository. Outcome measures, including measures of motor impairment, cognition and language, are assessed at 5, 15, 30, and 90 days post-stroke. The primary outcome measure is the upper extremity Fugl-Meyer score. Control participants are matched for age +/− 1 yr, race, gender, cardiovascular comorbities, and statin use through a computer algorithm that screens the entire MedStar electronic health record (EHR). Control participants provide 2 fasting blood samples one month apart. Once all samples are collected and sent for LCMS analysis, logistic regression analysis will identify potential metabolite biomarkers by comparing participants with good recovery to those with poor recovery as well as stroke participants to controls. RESULTS/ANTICIPATED RESULTS: To date, forty stroke participants have enrolled from 4 acute care hospitals in the Washington, DC metro region and completed all study procedures. Twenty stroke participants either dropped out or were withdrawn due to other medical concerns. Stroke patients ended up at a variety of venues following their acute hospitalization including the acute rehabilitation hospital, skilled nursing facilities, and home. We learned to overcome these logistical challenges by traveling to wherever the patients were sent and notifying medical providers of their study participation. In rare cases we have paid to transport patients from skilled nursing facilities to the clinic, which has reduced dropouts. In addition to the stroke participants, we have enrolled 7 healthy control participants using the EHR screening algorithm. DISCUSSION/SIGNIFICANCE OF IMPACT: Performing a longitudinal study in the early recovery phase following stroke is logistically challenging, but feasible. Difficulty in identifying participants with isolated motor impairment requires added effort to eliminate dropouts. Screening the EHR is an effective method to identify matched controls. Future metabolomics analysis of stored blood samples holds promise to identify biomarkers of stroke recovery and neural repair.

OBJECTIVES/GOALS: The training needs of clinical & translational scientists are evolving. Implementation of new curriculum content requires assessment of need, fit with current curriculum, incentives and barriers to implementation. We used implementation science methods to plan the implementation of a dissemination and implementation science training toolkit. METHODS/STUDY POPULATION: The Tufts Clinical & Translational Science (CTS) Graduate Program is the training core of the Tufts CTSI and its associated TL1. To assess barriers and facilitators to implementing the University of Washington’s Dissemination and Implementation (D&I) Science Toolkit for TL1 Programs into the Tufts CTS Graduate Program, we used the Consolidated Framework for Implementation Research (CFIR). We developed an interview guide based on CFIR constructs or categories of factors that might influence implementation of the D&I Toolkit and interviewed 8 stakeholders including leadership, trainees and faculty. We compared D&I Toolkit content to the current curriculum on stakeholder engagement (SE) and research study quality improvement (QI) to identify how the D&I Toolkit could be adapted to fit trainee and program needs. RESULTS/ANTICIPATED RESULTS: Stakeholder interviews and review of the current curriculum identified facilitators and barriers to implementing the D&I Toolkit in the Tufts CTS Graduate Program. The program has a strong SE focus and fosters the use of research study QI methods. Interviews identified student and faculty desire to implement the D&I content yet time constraints and overlap with the current curriculum were identified. The following plan to adapt the D&I Toolkit content to fit contextual factors was developed and executed: 1. Adapt didactic content to fit into a 90-min session. 2. Highlight the synergy between the current SE curriculum and the role of stakeholders in D&I science. 3. Modify course materials to highlight examples of disseminating local research results. 4. Limit the experiential learning component to a SE plan. DISCUSSION/SIGNIFICANCE: TL1 training programs must balance competing demands in selecting curriculum content. The CFIR framework was used to systematically assess implementation barriers by engaging program faculty, students and leaders to identify implementation strategies. This process could be useful when evaluating the addition of other educational content.

Participants in the leadership breakout session at the Clinical Translational Science Awards (CTSA) virtual 2020 conference discussed and ranked six recommendations in terms of feasibility, impact, and priority for advancing Diversity, Equity, and Inclusion (DEI) efforts to elevate underrepresented populations to leadership positions in CTSAs and their broader institutions. A thematic analysis of chat and polling data identified challenges and opportunities to achieve DEI goals, with the three most promising recommendations as: cross-institutional Principal Investigator (P.I.) action-learning workgroups, transparent policies for recruiting and promoting underrepresented minorities (URM) leadership, and a clear succession plan to nurture and elevate URM leaders. Suggestions are made to improve DEI in CTSA leadership and allow for greater representation in the translational science field.

OBJECTIVES/GOALS: The New Jersey Alliance for Clinical and Translational Science (NJ ACTS) interest in developing a comprehensive education and training program for enhancing the quality, efficacy, and safety of the clinical research enterprise led to the release of a survey distributed nationwide to assess initiatives in the translational science workforce. METHODS/STUDY POPULATION: Twenty-one hubs responded to the survey and data was exported from REDCap to Excel. Respondent demographics were categorized to formalize roles and data was categorically divided into sections based on training type (engagement, basic, postgraduate etc…) utilizing conditional formatting. The limitation in this survey was a branching logic defect aligned with questions on competency tool usage and the roles that they play which led to only six hubs having the advantage to respond to all questions. RESULTS/ANTICIPATED RESULTS: Summary findings showed that the majority of respondents for the survey (30%) were the Director of Operations’. Further; the Joint Task Force (JTF) domain Scientific Concepts and Research Design’ was the most preferred Hard Skill (81%) while the least preferred was Investigational Product Development and Regulation’ (29%). In spite of only six hubs receiving the short competency assessment; 50% of those hubs stated they would utilize ‘In House Assessment Tools’ and 83% stated they used the tools to ‘Develop Personalized Training Plans’. The assessment of this Inventory was indeed necessary to identify trends in available trainings across the CTSA consortium. DISCUSSION/SIGNIFICANCE: The internal cross training catalogue will help to develop an infrastructure for the NJ ACTS community to work along with other CTSA hubs while creating comprehensive clinical research training initiatives and programs.

OBJECTIVES/SPECIFIC AIMS: The objective of this analysis was to characterize the impact of stress, both early life and chronic, on intravenous alcohol self-administration (IV-ASA) in healthy non-dependent drinkers using the Computer-Assisted Infusion System (CAIS). Personality measures also have shown to impact drinking behavior, particularly impulsivity. Few studies have assessed the impact of stress and impulsivity on drinking behaviors in a non-dependent population. METHODS/STUDY POPULATION: Healthy non-dependent drinkers (n=28) completed a CAIS session, where they push a button adlib to self-administer standardized IV alcohol infusions. Participants completed the Cumulative Chronic Stress interview and the Early Life Stress Questionnaire (ELSQ) for stress measures. The Cumulative Chronic Stress interview was broken up into 4 sections: major life events, life traumas, recent life events, and chronic stressors. The number of endorsed events was added up to create 4 separate scores. Subjective response and craving measures were collected serially using the Drug Effects Questionnaire (DEQ) and Alcohol Urge Questionnaire (AUQ). The Impaired Control Scale (ICS) assessed failed control over recent drinking in the past 6 months. Impulsivity was assessed using the NEO personality inventory, which included the N-impulsive sub-facet, as well as the UPPS-P Impulsive Behavior Scale. RESULTS/ANTICIPATED RESULTS: Results showed early life stress events (ELSQ) are related to more chronic stressors in the cumulative chronic stress interview (p=0.005). Participants with higher chronic stress scores showed lower subjective effects, as measured by the DEQ, following the priming exposure (p=0.036) but had more craving for alcohol as measured by the AUQ (p=0.009). A regression analysis showed the number of chronic stressful events predicted ICS failed attempts to control drinking (p=0.034), after covarying for sex. Participants with more chronic stressful events showed more impulsivity on the N-impulsivity measure (p=0.034) and the UPPS-P positive urgency measure (p=0.005). DISCUSSION/SIGNIFICANCE OF IMPACT: Non-dependent drinkers with more early life stress tend to have a higher number of chronic stressful events. More chronically stressful events were associated with feeling less effects of alcohol and higher craving for alcohol. Participants with more chronically stressful events also appear to have more failed attempts at controlling their drinking. Future analysis will assess for mediation and moderation of these factors. Chronically stressful events and impulsive behaviors could serve as important areas for intervention for better treatment outcomes for alcohol use disorders.

OBJECTIVES/GOALS: Research is an important aspect of many students’ training. However, most trainees do not complete a scholarly project, and formal research training is rarely included in a degree program’s curriculum. Thus, we developed an online, asynchronous series of modules to introduce trainees to multiple topics that are relevant to the conduct of research. METHODS/STUDY POPULATION: Research 101 was utilized by first year medical students and undergraduate students conducting mentored research projects at the University of Cincinnati College of Medicine. Students’ knowledge, confidence, and satisfaction were assessed using pre- and post-module surveys with 5-point Likert scaled questions, open-ended text responses, and a final quiz. RESULTS/ANTICIPATED RESULTS: Pre-module survey results showed that learners were most confident with the Conducting a literature search and Race and racism in medicine modules and least confident with the Submitting an Institutional Review Board (IRB) protocol at UC module. Post-module survey responses were significantly increased compared to pre-module results for all modules and questions (p < 0.0001). The response to The content of this module met my needs was endorsed across all modules (84.9% yes responses). A final quiz of 25 multiple choice questions covering content from all required modules was completed by 92 students who had a median score of 21 (range: 15 to 25). DISCUSSION/SIGNIFICANCE: These data demonstrate significant learning resulting from completion of Research 101, as post-module survey scores were significantly higher than pre-module survey scores for all modules and questions. Final quiz scores were positive but also highlighted opportunity for additional student learning and will guide evolution of future modules.

OBJECTIVES/SPECIFIC AIMS: Our goal was to compare the ventriculo-arterial coupling and left ventricular mechanical work of patients with systolic and diastolic heart failure (SHF and DHF). METHODS/STUDY POPULATION: Patients with New York Heart Association Functional Class II-III HF symptoms were included. SHF was defined as left ventricular (LV) ejection fraction<50% and DHF as >50%. Analysis of the fingertip arterial blood pressure tracing captured with a finger plethysmography cuff according to device-specific algorithms provided brachial artery blood pressure and stroke volume. LV end-systolic volume was measured separately via transthoracic echocardiography. Arterial elastance (Ea), a measure of pulsatile and nonpulsatile LV afterload, was calculated as LV end-systolic pressure (ESP)/end-diastolic volume. End-systolic elastance (Ees), a measure of load-independent LV contractility, was calculated as LV ESP/end-systolic volume. Ventriculo-arterial coupling (VAC) ratio was defined as Ea/Ees. Stroke work (SWI) was calculated as stroke volume index×LV end-systolic pressure×0.0136 and potential energy index (PEI) as 1/2×(LV end-systolic volume×LV end-systolic pressure×0.0136). Total work index (TWI) was the sum of SWI+PEI. RESULTS/ANTICIPATED RESULTS: Patients with SHF (n=52) and DHF (n=29) were evaluated. Median (IQR) age was 57 (51–64) years. There were 48 (58%) and 59 (71%) patients were male and African American, respectively. Cardiac index was 2.8 (2.2–3.2) L/minute and 3.0 (2.8–3.3) L/minute in SHF and DHF, respectively (p=0.12). Self-reported activity levels (Duke Activity Status Index, p=0.48) and heart failure symptoms (Minnesota Living with Heart Failure Questionnaire, p=0.55) were not different between SHF and DHF. Ea was significantly lower in DHF compared with SHF patients [1.3 (1.2–1.6) vs. 1.7 (1.4–2.0) mmHg; p<0.001] whereas Ees was higher in DHF vs. SHF [2.8 (2.1–3.1) vs. 0.9 (0.7-1.3) mmHg; p<0.001). VAC was 1.8 (1.3–2.8) in SHF Versus 0.5 (0.4–0.7) in DHF (p<0.001). Compared with SHF, DHF patients had higher SWI [71 (57–83) vs. 48 (39–68) gm×m; p<0.001) and lower PEI [19 (12–26) vs. 44 (36–57) gm×m; p<0.001]. TWI did not differ between SHF and DHF (p=0.14). Work efficiency was higher in DHF than SHF [0.80 (0.74–0.84) vs. 0.53 (0.46–0.64); p<0.001]. DISCUSSION/SIGNIFICANCE OF IMPACT: The results underscore the differences in pathophysiology between SHF and DHF patients with similar symptom burden and exercise capacity. These results highlight the difference in myocardial energy utilization between SHF and DHF.

OBJECTIVES/SPECIFIC AIMS: Persons living with HIV (PLWH) are at increased risk for fragility bone disease. Current osteoporosis screening guidelines do not account for HIV status, and clinical risk assessment tools are not sensitive in PLWH. We examined the value of traditional osteoporosis risk factors, HIV-specific indices, and bone turnover biomarkers in predicting low bone mineral density (BMD) in PLWH. METHODS/STUDY POPULATION: Demographic and clinical characteristics, dual energy x-ray absorptiometry (DXA)-derived BMD, HIV indices (viral load, CD4 count, antiretroviral therapy [ART]), and biomarkers of bone turnover (C-terminal telopeptide of collagen [CTx], osteocalcin [OCN]) were evaluated in a cross-sectional analysis of PLWH (n=248) and HIV- controls (n=183). The primary outcome was low BMD, defined as osteopenia or osteoporosis by WHO criteria. Multivariable logistic and modified Poisson regression models were used to assess associations between low BMD and covariates of interest. RESULTS/ANTICIPATED RESULTS: Overall, median age was 44 years, 48% were male, 88% were black, median body mass index (BMI) was 28 kg/m2, 72% smoked cigarettes, and 53% used alcohol; characteristics did not differ by HIV status. PLWH had a mean CD4 of 408 cells/mm3, 55% were ART-naïve, and 45% had viral suppression on ART. Overall, 25% (109/431) had low BMD, including 31% of PLWH compared to 16% of HIV- controls. In multivariable models, HIV was significantly associated with low BMD (aOR 2.46, 95%CI 1.39-4.34; aRR 1.90, 95%CI 1.18-3.07). Adjusting for HIV, three traditional risks– age, race, and BMI– were independently associated with low BMD in the full cohort. However, bone turnover markers, CTx and OCN, were better able to discriminate low vs. normal BMD in PLWH compared to HIV- controls. In PLWH, mean serum CTx was 23% higher in low vs. normal BMD (mean CTx difference=0.06 ug/mL); in HIV- controls, no association with BMD was observed (mean CTx difference=0 ug/mL). In PLWH, mean serum OCN was 38% higher in those with low vs. normal BMD (mean OCN difference=2.48 ug/mL); in HIV- controls, mean serum OCN was only 16% higher in those with low vs. normal BMD (mean OCN difference=1.08 ug/mL). DISCUSSION/SIGNIFICANCE OF IMPACT: In PLWH as opposed to HIV- controls, serum biomarkers reflecting a high bone turnover state, may discriminate individuals with low versus normal BMD. Because changes in biomarkers precede changes in BMD, these markers should be explored further either alone or in combination with traditional risk assessment tools to improve early screening for osteoporosis in PLWH.

ABSTRACT IMPACT: A short treatment of 8 obese postmenopausal women with conjugated estrogens and bazedoxifene does not alter insulin sensitivity or ectopic fat but increases serum markers of hepatic de novo lipogenesis and production of triacylglycerides. OBJECTIVES/GOALS: Combining conjugated estrogens (CE) with the selective estrogen receptor modulator bazedoxifene (BZA) is a novel, orally-administered menopausal therapy. We investigated the effect of CE/BZA on insulin sensitivity, energy metabolism, and serum metabolome in postmenopausal women with obesity. METHODS/STUDY POPULATION: We conducted a randomized, double-blind, crossover pilot trial, testing the effect of CE/BZA on cardiometabolic health in postmenopausal women. Eight postmenopausal women (age 50-60 y, BMI 30-40 kg/m2) were randomization to an 8-week CE/BZA or placebo treatment separated by an 8-week washout period [NCT02274571]. The primary outcome was insulin sensitivity (hyperinsulinemic-euglycemic clamp), while secondary outcomes included body composition (DXA); resting metabolic rate (RMR); substrate oxidation (indirect calorimetry); ectopic lipids (1H-MRS); fat cell size, adipose and skeletal muscle gene expression (biopsies); inflammatory markers; and serum metabolome (LC/MS). RESULTS/ANTICIPATED RESULTS: CE/BZA had no effect on insulin sensitivity, body composition, ectopic fat, or substrate oxidation, but resulted in a non-significant increase in RMR (basal: p=0.06; high-dose clamp: p=0.08) compared to placebo. CE/BZA increased serum high-density lipoprotein cholesterol. CE/BZA also increased serum diacylglycerol (DAG) and triacylglycerol (TAG) species containing long-chain saturated, mono- and polyunsaturated fatty acids (FAs), and decreased long-chain acylcarnitines. These findings possibly reflect increased hepatic de novo FA synthesis and esterification into TAGs, and decreased FA oxidation, respectively (p<0.05). CE/BZA increased serum phosphatidylcholines, phosphatidylethanolamines, ceramides, and sphingomyelins, possibly reflecting the increase in lipoproteins (p<0.05). DISCUSSION/SIGNIFICANCE OF FINDINGS: A short treatment of postmenopausal women with CE/BZA did not alter insulin action or ectopic fat, but increased markers of hepatic de novo lipogenesis and TAG production. Study limitations include a small sample size and short treatment period. A larger, fully powered study is needed to validate the potential metabolic benefit of combining CE with BZA.

OBJECTIVES/GOALS: Whole-genome viral sequencing is vital to inform public health and study evolution. Arboviruses evolve in vectors, reservoir hosts, and humans, and require surveillance at all points. We developed a new rigorous method of sequencing that captures whole viral genomes in field-collected and clinical samples. METHODS/STUDY POPULATION: ClickSeq is a novel method of Next Generation Sequencing (NGS) library synthesis using azido-nucleotides to terminate reverse transcription. The cDNA generated can be ligated to sequencing and indexing primers at room temperature using copper (Cu I) and vitamin C. With this approach, we designed primers located ~250 bp apart along the genomes of the arboviruses Chikungunya 37797, Zika Dakar, Yellow Fever Asibi, Dengue serotype 2, West Nile 385-99, and St. Louis Encephalitis Virus (SLEV) clade II. We tested this method with varying viral titers: lab-infected mosquito pools, field-collected mosquito pools from a Texas West Nile and SLEV outbreak, and patient isolates from a Pakistani CHIKV outbreak. The cDNA was sequenced in the UTMB NGS Core and aligned using bowtie. RESULTS/ANTICIPATED RESULTS: The use of a single protocol to capture whole viral genomes including UTRs for multiple viruses from different sample collection styles is ideal for arboviruses. Primers for multiple viruses were pooled and used to sequence mosquito pools. The Tiled ClickSeq method captured whole viral genomes without the need for host depletion. UTRs were captured even when the viral strain used for primer design differed from the resulting strain. Discreet variants were captured in both the hypervariable nsP3 region and the UTR in the patient isolates from the CHIKV outbreak compared to the 2017 outbreak. Texas WNV and SLEV outbreaks are now defined from the 2020 outbreak and can be further tracked to update public health measures and understand viral evolution. DISCUSSION/SIGNIFICANCE: UTRs impact both human and mosquito fitness, leading to further outbreaks. Tiled ClickSeq aims to capture whole viral genomes with a method and cost that can be implemented by public health researchers to understand disease evolution as it happens to update both public health and basic virology to the effects of evolution on arboviruses.

OBJECTIVES/SPECIFIC AIMS: Our study’s primary aim is to determine if there is an association between cancer-related pain among patients who underwent major elective procedures and postoperative opioid overdose. In addition, the relationship between cancer-related pain in this population and inpatient mortality, total hospital charge and length of stay was assessed. METHODS/STUDY POPULATION: Our study sample consisted of adults 18 years and older who had at least one of eight elective procedures. Data was obtained from the National Inpatient Sample (NIS). Variables were identified using ICD-9 codes. Our primary predictor was cancer-related pain while our primary outcome was opioid overdose. Secondary outcomes were inpatient mortality, length of stay and total charge. Propensity-matched regression models were employed in assessing the association between cancer-related pain and outcomes of interest. RESULTS/ANTICIPATED RESULTS: Among 4,085,355 selected patients, 0.8% (n = 2,665) had cancer-related pain while 99.92% (n = 4,082,690) had no diagnosis of cancer-related pain. All subjects with cancer-related pain (n = 2,665) were successfully matched to subjects with no diagnosis of cancer-related pain in a 1:5 ratio yielding 13,325 controls. Patients with cancer-related pain had significantly higher odds of opioid overdose (aOR 4.82 [95% CI [2.68-8.67]; p-value <0.0001) and inpatient mortality (aOR 1.39[1.11-1.74]; p-value 0.0043). Patients with cancer-related pain were also likely to stay significantly longer in the hospital (12.76 days vs. 7.88 days) with significantly higher total hospital charges ($140,220 vs. $88,316). DISCUSSION/SIGNIFICANCE OF IMPACT: Pain is a common complication of cancer pathogenesis, diagnosis or treatment. Though a rare outcome, opioid overdose could lead to undesirable outcomes. Cancer patients undergo invasive diagnostic and therapeutic procedures as part of their cancer management or for conditions not related to their primary cancer diagnosis. Safety measures including alternatives to opioids are recommended to prevent the poor clinical outcomes and higher healthcare utilization indices associated with opioid overdose in this population.

OBJECTIVES/SPECIFIC AIMS: The food reward circuitry regulates hedonic eating especially in relation to palatable hypercaloric foods, which can lead to chronic overeating and consequent overweight and obesity. Evidence supports that there is considerable overlap within the brain reward circuitry between palatable hypercaloric food intake and substance addiction. The goal of this study was to identify associations between addiction-related genes and body mass index. We hypothesized that addiction-related genes potentially participate in the food reward circuitry if they are associated with obesity traits. METHODS/STUDY POPULATION: A secondary analysis was conducted with 1093 African American adolescents and young adults from the New Mother’s Study. Anthropometric, genetic, demographic and lifestyle measurements were available at the 18-year follow-up assessments. A total of 1350 single nucleotide polymorphisms mapped to 127 addiction-related genes were assessed. A total of 186 ancestry informative markers were used to adjust for population stratification. Generalized estimating equation models were used to identify genetic associations, including additive, dominant, and recessive models, and control for correlations within families. RESULTS/ANTICIPATED RESULTS: The participants ranged from 15 to 23 years of age. Of them, 42.7% were overweight or obese. Significant associations with body mass index were identified for 13 single nucleotide polymorphisms mapped to 11 addiction-related genes, including LEP (p 0.027–<0.001). Most of these genes are involved in dopaminergic, opioidergic, serotonergic pathways, and stress. DISCUSSION/SIGNIFICANCE OF IMPACT: Our results support the role of dopaminergic and opioidergic pathways in the food reward circuitry, and suggest a potential involvement of serotonergic pathways and genes related to stress in the food reward circuitry. Further investigation of the identified genes will facilitate delineation and understanding of the brain food reward system and its relationship with obesity.

OBJECTIVES/SPECIFIC AIMS: Insufficient endogenous expression of suppressor of cytokine signaling 3 (SOCS3) with subsequent over-activation of its target, the transcription factor STAT3, has been associated with tumorigenesis and cancer development in the lung and other organs. We have observed that a “backup” source of SOCS3 in the lung, namely that secreted in microvesicles (MVs) by alveolar macrophages, is reduced in the bronchoalveolar lavage fluid (BALF) of KRAS mutant mice harboring lung tumors. Here we sought to evaluate levels of SOCS3 in BALF of a cohort of non-small cell lung cancer (NSCLC) patients and to test the effects of vesicular SOCS3 administration on tumor cell transformation and function as potential therapeutic strategy. METHODS/STUDY POPULATION: In total, 22 BALF samples were obtained from healthy volunteers (n=11) as well as patients undergoing diagnostic bronchoscopies for suspected lung cancer (n=11). SOCS3 levels in the BALF were determined by ELISA after brief sonication to disrupt vesicles. In vitro experiments utilized the human adenocarcinoma cell line (A549) or human G12V mutant KRAS-expressing rat lung epithelial cells (RLE-G12V). Proliferation, Fas ligand (FasL)-induced apoptosis, and chemical transformation with N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) or cigarette smoke extract (CSE) were assessed by CyQuant assay, annexin V staining, and soft agar assay, respectively. For SOCS3 rescue, epithelial cells were treated with natural alveolar macrophages-derived MVs (isolated via ultracentrifugation) or synthetic unilamellar liposomes containing human recombinant SOCS3 for at least 1 hour before assay. RESULTS/ANTICIPATED RESULTS: SOCS3 levels were significantly reduced in BALF samples of patients determined to have NSCLC as compared with healthy volunteers (186.6±26.74 vs. 395.6±74.31 pg/mL, p=0.015, n=11). Addition of exogenous SOCS3-containing liposomes had the capacity to significantly inhibit MNNG and cigarette smoke extract-induced transformation and colony formation in soft agar. Exogenous SOCS3 provided in liposomes or in natural MVs significantly induced apoptosis (both in the presence and absence of FasL) and inhibited basal proliferation of A549 cells. DISCUSSION/SIGNIFICANCE OF IMPACT: These data identified a novel dysregulation of immune surveillance in the form of decreased SOCS3 secretion in the tumor-bearing lung that may contribute to tumorigenesis via sustained STAT3 activation. Future studies will focus on the mechanism underlying this defect and whether rescuing SOCS3 secretion can inhibit cancer progression in vivo.