OBJECTIVES/GOALS: To assess researchers’ experiences working with community advisory boards (CABs) and how community member/patient stakeholder feedback impacted the research. METHODS/STUDY POPULATION: Researchers (N = 34) who presented their research to a Mayo Clinic CAB (at MN, AZ, or FL) from 2014-2017 were invited to participate in an interview in-person or by phone averaging 10-15 min. Researchers were asked “In what ways did the feedback you received from the CAB influence your research?” The validated structured 7-item interview included domains assessing potential influence that CABs had on the research: 1) Pre-research (e.g., generated ideas), 2) Infrastructure (e.g., budget preparation), 3) Research design, 4) Implementation (e.g., research recruitment), 5) Analysis, 6) Dissemination, and 7) Post-research (e.g., assist in formulating next steps). RESULTS/ANTICIPATED RESULTS: 17 interviews were completed (8 no longer at Mayo, 9 no response). Researchers presented their study to a CAB a mean of 4 years (range 3-5) before the interview. Researchers reported that the CAB had influenced their research in the following domains: 24% in pre-research, 24% infrastructure, 41% study design, 41% implementation, 6% analysis, 24% dissemination, and 18% for post-research activities. The mean total score was = 1.8 (SD = 1.7, range 0-6), of a possible range of 0-7. DISCUSSION/SIGNIFICANCE OF IMPACT: Impact of CAB feedback on the research was moderate. Ways to enhance impact could include follow-up with researchers and CAB members.

OBJECTIVES/GOALS: Achieving therapy for hepatocellular carcinoma (HCC) involves navigating through a complex cascade of care. Non-HCC cancer mortality has been associated with social determinants of health outside of cancer specific risk. Our objective is to explore the impact of social determinants on HCC outcomes. METHODS/STUDY POPULATION: Patients with HCC were enrolled from 3 hospitals form June, 1 2019 to December 1, 2019. A chart review was done to collect information on liver disease severity and cancer stage. Patients were interviewed to collect information on the following: 1) socioeconomic status (income, education, insurance status, and employment status), 2) literacy (Rapid Estimate of Adult Literacy in Medicine (REALM-R) and Brief Health Literacy Screening Tool (BREIF)), 3) social support (Patent Reported Outcome Measurement Information System (PROMIS) instrumental and information support tool), 4) quality of life (PROMIS global and mental health tool), 5) substance abuse, and 6) linkage to care. RESULTS/ANTICIPATED RESULTS: Data compiled on the social determinants of health revealed (n = 35): 1) 60.0% of patients had incomes below $30,000 per year, 60.0% of patients had not gone past high school for education, and 8.6% had full time employment, 2) the average BREIF score was 10.3 (range 3-15)(4-12 indicate limited literacy). The average REALM-R score was 5.5 (range 0-8) (<6 indicate at risk for poor literacy), 3) patients had strong instrumental (T score 61.4±7.1) and information social support (T score 64.6±4.7) (mean T scores calibrated to a general population mean of 50), 4) patients had poor mental (T score 43.7 ±6.5) and physical quality of life (T score 46.6 ±9.9), 5) 25.7% of patients reported alcohol use in the past 90 days 6) 80.0% of patients reported that their doctor had spoken to them about liver transplantation. DISCUSSION/SIGNIFICANCE OF IMPACT: This patient population was well linked to care with good social support. However their literacy, socioeconomic status, mental and global health was poor and substance use history complex. Continued follow up of this cohort is planned to determine how these factors might impact their ability to navigate through the care cascade as well as survival.

OBJECTIVES/GOALS: Undergraduates experience food insecurity at rates 21% higher than the general population. Because professional students have been omitted from these studies, the goal of this project is to determine the prevalence of food insecurity among medical students at one academic institution. METHODS/STUDY POPULATION: A cross-sectional research design was used to quantify the food insecurity status of medical students at the University of Utah. The USDA’s validated 6-item Food Security Survey Module was distributed via email to all currently matriculated medical students. Student’s responses were anonymous but questions about gender and age were included. Respondents (N = 200) were scored per the module as food secure, food insecure, or very low food security. RESULTS/ANTICIPATED RESULTS: Statistical analysis included frequencies and chi-square tests. Medical students (N = 166) showed 50.6% of respondents experienced food insecurity in the past 12 months, 16.3% experienced very low food security.While there were no significant relationships between food security status and gender or age, general trends did show divorced and separated students had higher food insecurity risk 82%. A similar study in 2014 surveyed undergraduates at the same location; 51% of respondents (N = 221) experienced food insecurity. While medical students experience food insecurity at rates much higher than the national average, prevalence is lower than undergraduates at the same institution. DISCUSSION/SIGNIFICANCE OF IMPACT: Burnout and suicide in medical training are at an all-time high; professional and academic pursuits are limited when physiological needs of food security are not being met. Study results suggest, 50% of respondents are food insecure. This should inform the development of supportive programs.

OBJECTIVES/GOALS: Asthma is a life-long, chronic lung disease that inflames and narrows the airways. Its effects on quality of life in children can be exacerbated. The goal of this study was to investigate the link between asthma, family structure and demographics and how it impacts quality of life in children. METHODS/STUDY POPULATION: We analyzed data from a cross sectional study of the 2016-2017 National Survey of Children’s Health, NSCH, to assess the relationship between determinants of health variables and the outcome variable of parent’s report of child’s perceived health status (quality of life). The study population was children under the age of 18. Data were analyzed using descriptive, bivariate analysis using Chi square, and multiple logistic regression of quality of life and family structure adjusting for confounding variables. RESULTS/ANTICIPATED RESULTS: The study included 5,687 children. Significant predictors of asthmatic children’s quality of life were severity of asthma, self-perceived mental/physical health status of adults, neighborhood safety for children (p <0.05). The interaction between family structure and asthma severity was significant indicating that asthma severity was an effect modifier. Among children with mild asthma, predictors of quality of life were self-perceived mental/physical health status of adults in the household, neighborhood safety of children, physical activity status of children (p<0.05). Among children with severe asthma, predictors were family structure and physical/mental health of adults (p<0.05). DISCUSSION/SIGNIFICANCE OF IMPACT: This study suggests children with severe asthma who are born to single mothers with lower parental reporting of physical/mental health status had a lower quality of life. A longitudinal study could be implemented to target these three measures to improve quality of life among these children. Also, a culturally adapted intervention involving community, parents, and providers is needed to improve the quality of life of the children with asthma.

OBJECTIVES/GOALS: Using a human-centered approach, IDEO, a nationally-renown human-centered design team, and Research Jam, Indiana CTSI’s patient engagement core, integrated and tailored complimentary programs to address the challenges of engaging mothers with opioid misuse around the time of birth. METHODS/STUDY POPULATION: Gathered data through focus groups, site visits, and one-on-one interviews with key stakeholders: mothers in opioid use recovery, peer recovery coaches, and other people living with or directly affected by opioid use disorder (OUD). RESULTS/ANTICIPATED RESULTS: Themes emerged around stigma (e.g., constant judgment, majority of interactions focused on addiction, addiction comes from bad choices), the healthcare system (e.g., healthcare system bias and stigma, misalignment of services and timing of need, no support for support network), and relating to recovery (very variable but generally ambiguous and uncertain process and outcomes, importance of peer recovery coaches, importance of community resources). Identified themes were used to create insights that informed the underlying concepts of an engagement strategy including support and resources for recovery coaches, and education materials for mothers with OUD. One of human-centered design’s strengths is iteration, and the materials created for this have yet to be tested and refined thoroughly to be meaningful and lasting interventions. DISCUSSION/SIGNIFICANCE OF IMPACT: Considerable insights into the lived experience of those experiencing OUD and those who support these individuals yielded tangible ways to test improved engagement and recruitment of women with OUD at the time of birth.

OBJECTIVES/GOALS:

1. 1. Identify patient-level factors associated with hemoglobin A1c reduction and sustained device use after 12months of participation in a diabetes and hypertension remote monitoring program

2. 2. Utilize qualitative methodology to characterize key barriers and facilitators to remote monitoring engagement

METHODS/STUDY POPULATION: All participants in statewide quality improvement initiative utilizing a cellular-enabled device with glucose and blood pressure monitoring capability will be included in quantitative analysis (N = 302 at baseline and N = 125 at 6 months at the time of analysis). We developed multilevel regression analyses to model factors associated with clinical outcome (hemoglobin A1c change) and transmission frequency over time. Focus groups and surveys will be conducted to identify barriers and facilitators to continued data transmission and hemoglobin A1c change over 12 months. Semi-structured interview guides are mapped to Wagner’s Chronic Care Model. RESULTS/ANTICIPATED RESULTS: Overall, program participation was associated with 1.8% and 1.3% A1c reduction at 6 (n = 302) and 12 months (n = 125). Regression models showed no association of age, gender, race, income, or insurance with hemoglobin A1c change. Modeling of patient factors associated with sustained transmission frequency or device use is ongoing. Patient focus groups and surveys are currently being scheduled and qualitative data will be analyzed using content analysis. After completing qualitative and quantitative data analyses independently, we will use graphical matrix configurations (“joint displays”) to synthesize findings. DISCUSSION/SIGNIFICANCE OF IMPACT: Our goal is to identify variables associated with the likelihood of patients to engage in and benefit from sustained remote monitoring. Results may inform health policy and guide recruitment approaches, implementation strategies, and methodologic design for future trials. CONFLICT OF INTEREST DESCRIPTION: The authors have no conflicts of interest or disclosures to report

OBJECTIVES/GOALS: Feeding preterm infants with mother’s own milk (MOM) lowers rates of sepsis, decreases necrotizing enterocolitis, and shortens hospital stay. Our objective is to determine whether a similar microbial diversity to MOM can be obtained when fresh or frozen MOM is inoculated in donor human milk (DHM). METHODS/STUDY POPULATION: Subjects included 12 mothers of infants born 100ml of MOM per day and were excluded if they had taken antibiotics within 3 days of the 1-time pumped MOM sample collection. MOM sample was divided into fresh (processed immediately) and frozen (−20°C) for 24h fractions. MOM was inoculated in DHM [referred to as refaunated milk (RM)] at 10% (RM10) and 30% (RM30) dilutions, then incubated at timepoints: 0h, 2h, 4h at 37°C. At each timepoint, total viable microbial cell counts were performed in differential or selective media along with future 16S rRNA sequencing. RESULTS/ANTICIPATED RESULTS: Microbiota expansion was detected in MOM, RM10 and RM30 over time whether fresh or frozen milk was used as the inoculum. Incubated fresh and frozen MOM had similar bacterial loads when tested on nutrient agar (10^5-10^6 CFU/mL), mannitol salt (10^6 CFU/mL), MacConkey (10^2-10^5 CFU/mL), blood agar (10^6 CFU/mL) and MRS (10^4 CFU/mL) plates. Based on these CFU counts, RM30 incubated for 2h and RM10 at 4h showed similar counts to that of MOM at 0h. DISCUSSION/SIGNIFICANCE OF IMPACT: RM, inoculated with fresh or frozen MOM, obtained a similar microbial count compared to MOM at 0h indicates that fresh or frozen MOM can inoculate DHM. 16s rRNA sequencing is ongoing. Future studies are needed to support an inoculation protocol to be used in clinical practice and human milk banking.

OBJECTIVES/GOALS: Early dysphagia detection reduces risk of pulmonary complications, length of hospital stay, and overall healthcare costs. The biggest limitation for early detection has been the lack of a sensitive, reliable, and noninvasive screening tool. The bedside swallow examination may miss silent aspiration in up to 40% of patients. The objective of this study is to evaluate if acoustic parameters can distinguish normal and wet voice in a canine laryngeal model. Ultimately, our goal is to establish whether the sensitivity of the bedside swallow examination can be augmented with the addition of an acoustic screen in humans. METHODS/STUDY POPULATION: Two excised canine larynges were used for laryngeal phonation simulations under six different conditions over 48 hours. Acoustic recordings were obtained while the larynges were in vibration at the phonation threshold pressure. Phonation was recorded under dry conditions and when the laryngeal introitius was covered with low viscosity glycerin (9.5cP) or high viscosity glycerin (950cP), as well as in adducted and abducted conditions. The latter mimics glottic insufficiency seen in presbylarynx or vocal fold paralysis. RESULTS/ANTICIPATED RESULTS: A total of 112 voice samples were generated and analyzed for pitch, sound pressure level (SPL), % shimmer, % jitter, relative average perturbation (RAP), and noise-to-harmonics using PRAAT software. A multivariate regression model showed that pitch, SPL, % shimmer, % jitter and RAP could significantly predict wetness in abducted conditions only. Could you please add numbers and p values? DISCUSSION/SIGNIFICANCE OF IMPACT: This pilot study indicates that classic acoustic perturbation measures distinguish the dry from the wet larynx only in glottic insufficiency condition in an ex vivo canine laryngeal model. Our next step is to study whether non-linear time series analysis and machine learning can differentiate dry and wet phonation in both adducted and abducted conditions in our animal model. CONFLICT OF INTEREST DESCRIPTION: Dr. Anais Rameau is a co-founder and Chief Executive Officer of MyophonX, a wearable device used to restore speech in patients with limited phonation capacity.

OBJECTIVES/GOALS: This study seeks to culture and characterize mucus-degrading microbes from the microbiome of inflammatory bowel disease (IBD) patients. METHODS/STUDY POPULATION: Stool will be collected from IBD patients and healthy first-degree relatives, then enriched for mucin-degrading microbes through growth on porcine rectal mucin. Dilution plating in both liquid and solid culture formats will be employed to isolate strains capable of growth on mucin. Cultures that are positive for mucin degradation will be identified with 16S rRNA sequencing; unique isolates will be genome sequenced and transcriptionally profiled on simple monosaccharides and mucin in order to identify putative mucin-degrading genes. The abundance of novel enzymes, pathways, and microbes will be compared in healthy and IBD patient populations using existing datasets in the literature. RESULTS/ANTICIPATED RESULTS: We expect to isolate previously uncultured mucin-degrading microbes, which will likely include new strains and possibly new species of bacteria. Through the transcriptomic characterization of mucin-degrading pathways, we will expand the lexicon of known mucin-degrading enzymes and pathways used by bacteria in the human colon. We expect mucin-degrading microbes to be more abundant and active in IBD patients compared to healthy controls. DISCUSSION/SIGNIFICANCE OF IMPACT: There is no cure for IBD and treatment relies heavily on suppressing a patient’s immune system. This research seeks to understand the contribution of the gut microbiota in the pathogenesis of IBD, which may lead to future therapeutic targets.

OBJECTIVES/GOALS: We study the association of adaptive decision-making, motor planning, and neuromuscular constraints, in children with hemiplegia. We examine how children scale motor decisions to body mechanics and the distance of a target while reaching in sitting/standing, and if they can recalibrate motor decisions to sudden changes in body mechanics. METHODS/STUDY POPULATION: Forty-five 6-12 year-olds with hemiplegia and 45 age/gender matched typically developing controls participate in clinical tests (i.e. balance, visual perceptual skills, etc.) and 3 experiments. Children “reach to tap” toward a target while sitting with both preferred and not preferred arms under three conditions: regular elbow extension siting and standing and elbow extension range reduced by 50% via a splint while sitting. Trials are easy, ambiguous, and difficult. Motor decisions are compared to abilities and motion sensors (IMUs) worn at wrist, arm, sternum and lumbar area, record biomechanical strategies children use under different decisions. Synchronized video analysis presents biomechanical strategies under different decisions. RESULTS/ANTICIPATED RESULTS: Data collection is still underway. A mixed models analysis is used to compare 2 (group: hemiplegic/typically developing) X 2 (arms: healthy/impaired & dominant/non dominant) X 3 (difficulty levels) the children’s decisions. Functional analysis is used to capture biomechanical strategies children use under different decisions and levels of difficulty. Exploration strategies are recorded relative to levels of difficulty. We will also compute correlations between affordance thresholds for all children and measures of sensation, range of motion, cognition and balance (in each posture). Lastly, a secondary analysis will compare behaviors of children with left/right hemisphere lesions, as they differ in spatial abilities. Preliminary results show that children with hemiplegia make errors with both their affected and unaffected side. DISCUSSION/SIGNIFICANCE OF IMPACT: Motor deficits in children with hemiplegia are the primary focus of treatments. Motor learning interventions focus on biomechanical deficits. Results from these studies expand the focus to planning and cognitive control issues underlying motor deficits.

OBJECTIVES/GOALS: An age-dependent restitution defect in our neonatal pig intestinal ischemia model is rescued by unknown factors within homogenized mucosa of weaned pigs. A postnatally maturing network of enteric glia regulates the epithelial barrier, so we aim to show rescue is due to replacement of glial factors. METHODS/STUDY POPULATION: Jejunal tissues from suckling or weaned pigs were assessed by RNAseq and processed for immunofluorescent histology and 3-D volume imaging. Jejunal ischemia was surgically induced in weaned pigs and injured mucosa was recovered ex vivo with or without the glial inhibitor fluoroacetate (FA) while monitoring transepithelial electrical resistance (TER). RESULTS/ANTICIPATED RESULTS: Ingenuity Pathways Analysis of RNAseq data revealed significant suppression of numerous pathways critical for epithelial wound healing in suckling pigs (Z-score <−2 for of nine key pathways). Volume imaging studies confirmed lower density (P≤0.05) and complexity of the subepithelial glial network in suckling pigs. Treatment with FA inhibited TER recovery (P<0.0001) and restitution (P<0.05) in weaned pigs, mimicking the suckling pig phenotype and supporting glia as an important regulator of restitution in our model. DISCUSSION/SIGNIFICANCE OF IMPACT: These findings provide important evidence that a developing glial network may be critical to the postnatal development of intestinal barrier repair mechanisms. Ongoing work will explore glial-epithelial interactions in vitro to further define postnatal development of barrier repair.

OBJECTIVES/GOALS: Dysfunction of the autonomic nervous system (ANS) may be important in both depression and coronary artery disease (CAD). A novel heart rate variability (HRV) metric, Dyx, may be a be a potentially useful tool to study ANS dysfunction in these diseases. We propose that ANS dysfunction, measured by decreased Dyx, will associate with both depression and obstructive CAD. METHODS/STUDY POPULATION: We included participants undergoing coronary angiography for suspected CAD. Depressive symptoms were assessed with the Patient Health Questionnaire-9 (PHQ-9). HRV data were collected continuously on participants before catheterization using a new ECG patch (VivaLNK). We assessed HRV by Dyx (primary) and high and low frequency power, multiscale entropy, and deceleration capacity. Two-sample t-tests and logistic regressions (with adjustment for age and sex) were used to study the difference in HRV (before cardiac catheterization) between those with high versus low depressive burden (PHQ-9 ≥ 10), and in those with versus without obstructive CAD (>70% stenosis). RESULTS/ANTICIPATED RESULTS: We assessed 30 individuals with mean (SD) age 62.4 (13.2); 7.1% were female and 15.4% were black. Mean Dyx in high depressive symptoms (N = 21, 70%) was 1.8 (0.2) and in none-low depressive symptoms (N = 7, 23%) was 2.2 (0.2). Differences were also observed for high frequency (HF) (4.4 (1.1) vs. 6.0 (1.4)) and deceleration capacity (−4.2 (2.1) vs. −10.7 (8.5)). Mean Dyx in obstructive CAD (N = 17, 57%) and non-obstructive CAD (N = 10, 33%) was 1.7 (0.6) and 2.6 (1.2) respectively. Differences were seen with sample entropy (1.2 (0.2) vs. 1.5 (0.2)). Every 1 unit of log(HF) had an odds ratio = 0.14 (95% CI 0.06 – 0.36) for depression. DISCUSSION/SIGNIFICANCE OF IMPACT: ANS dysfunction, measured by HRV, associates with both depression and obstructive CAD. Autonomic ECG markers may play an important role in assessing brain-heart pathology, and may be useful to study the interaction between depression and CAD.

OBJECTIVES/GOALS: Current surgical flap and replantation monitoring techniques have limitations in detecting the pathologic state, calibration and cost-to-patient issues. Our hypothesis is that novel implantable oxygen sensors can provide a more efficient, accurate, and reliable monitoring of tissue oxygenation. METHODS/STUDY POPULATION: Experimental sensors were used with an exogenous remote used as a reader once implanted (Fig. 1) A rat tissue perfusion model with three regions of an SIEA flap as well as into adjacent control sites was made (Tip, Middle, and Base) Blood flow was greatest at the base, diminishing towards the Tip, thus creating a perfusion gradient. Changes in tissue oxygen tension PO2 were estimated by the steady-state fluorescence of the optical sensors using an IVIS imaging system. The sensors were used to collect data from days 0, 3, and 7 as a reading of Tissue Oxygen Tension (TOT) with ANOVA used to assess for statistical significance in blood oxygen data with respect to relative perfusion status. RESULTS/ANTICIPATED RESULTS: Inspired FiO2 was decreased from 100% to 12% with a corresponding change in the TOT readings from all sensors. (Fig. 2) The tip portion of the flap demonstrated the most profound detection of tissue necrosis, with the middle demonstrating the second most necrosis and the base demonstrating the least with correlating TOT sensor readings. (Fig. 3) Acute vascular compromise of the feeding blood vessels in the pedicle was immediately detected within 70 seconds (*p<0.05). (Fig. 4) DISCUSSION/SIGNIFICANCE OF IMPACT: This study introduces and validates a recent technique to monitor acute vascular occlusion, flap viability, and necrosis in the immediate postoperative period in a validated rodent model. Future directions of this novel technology will aim to reproduce these findings in clinical feasibility studies.

OBJECTIVES/GOALS: NASH increases the risk of cirrhosis and liver cancer. High-fat diets increase CCK levels and CCK receptors have been identified on fibroblasts and immune cells. We hypothesized that CCK receptor blockade could prevent NASH by altering the hepatic microenvironment and macrophage activation. METHODS/STUDY POPULATION: Female mice were fed a Choline Deficient Ethionine supplemented (CDE) saturated fat diet or control high-fat diet for 18 weeks. Mice in each group were treated with a CCK receptor antagonist, proglumide (0.1 mg/ml) in the drinking water or regular water. Resected livers were stained for H&E for features of NASH and F4/80 for macrophages analysis. Liver RNA was evaluated for the expression of cytokines and chemokines using an 84-gene Profiler array (Qiagen). Oxidative stress was analyzed by qRT-PCR for heat shock proteins (HSPs) 27, 60, 70 and 90 and for glutathione by a fluorometric assay. Differences in CDE fed and CDE/proglumide-treated mouse livers were evaluated. RESULTS/ANTICIPATED RESULTS: Livers from mice on the CDE diet displayed histologic features of NASH that were prevented by proglumide. Cytokines and chemokines expression, especially CCL20 and CCL2, were increased in the CDE fed mice and these levels were reduced greater than 20-fold with proglumide. Infiltration of F4/80+ macrophages was markedly increased in the CDE livers and these were reduced by > 50% (p < 0.0001) with proglumide. RNA expression of HSP70 (p = 0.006) and HSP27 (p = 0.011) were reduced with proglumide. Hepatic glutathione concentration more than doubled in the CDE/proglumide treated mice compared to CDE mice. CCK-B receptor expression increased in the CDE-fed mouse livers compared to controls. DISCUSSION/SIGNIFICANCE OF IMPACT: CCK receptor blockade decreases NASH by reducing hepatic macrophages, oxidative stress, and blocking inflammatory cytokines and chemokines. This data supports our novel hypothesis that CCK receptors play a role in NASH and proglumide may provide an innovative treatment for this condition.

OBJECTIVES/GOALS: Experimental pain testing is used to identify changes in nociceptive processing and outcomes with intervention. This study investigated exercise induced changes in experimental pain and self-reported pain intensity after an acute bout of exercise in participants with fibromyalgia. METHODS/STUDY POPULATION: Ten females with fibromyalgia (55±10yr) were familiarized to study procedures and underwent submaximal (20% maximal voluntary contraction) intermittent eccentric muscle contractions isolated to the right arm for 10-minutes. Self-reported pain intensity (0-10 numerical pain rating scale [NPRS]) of the exercising arm was measured before, during, and after exercise; whole-body pain intensity was measured before and after exercise. Experimental pain testing included measurement of pressure pain thresholds (kPa [PPTs]); temporal summation (TS) of pressure pain with a constant mechanical pressure; and TS of punctate pressure with repeated application of monofilaments before and after exercise. RESULTS/ANTICIPATED RESULTS: Participants reported minimal to moderate arm pain (3.1±2.1) during exercise. Following exercise, arm pain and whole-body pain significantly increased (3.1±2.2 and 1.6±0.5, respectively) [p<0.05]. No change occurred with PPTs at the bicep (138.9±49.5 to 142.8±55.3), PPTs at the quad (212.0±105.4 to 228.1±100.0), TS of mechanical pressure pain (7.6±2.1 to 7.9±1.5), TS of punctate pressure pain at the bicep (2.6±1.7 to 3.0±1.5), and TS of punctate pressure pain at the quad (3.6±1.5 to 3.7±1.4) before to after exercise respectively [p>0.05]. The change in self-reported arm and whole-body pain did not correlate with the change in experimental pain testing. DISCUSSION/SIGNIFICANCE OF IMPACT: In people with fibromyalgia, there is no relation between self-reported clinical pain and experimental pain following a single exercise session. Further research should identify the influence of exercise training on pain perception and if experimental pain testing translates to clinical insight.

OBJECTIVES/GOALS: This study uses diffusion kurtosis imaging (DKI) to investigate the structural profiles of basal ganglia (BG) motor circuitry in Vascular Parkinsonism (VP), Parkinson’s disease (PD), and healthy aging controls (HC). VP is a clinical diagnosis of lower body predominant parkinsonism without significant benefit from levodopa. VP is distinct from PD, yet the concept of VP remains debated due to the inability of prior studies to identify specific causative changes. One reason for this may be limitations in measuring intricate BG connectivity in vivo. Given the predominant lower body parkinsonism symptoms in VP, we hypothesized that VP would be associated with decreased connectivity specifically within the BG motor loop. METHODS/STUDY POPULATION: We obtained DKI brain imaging in subjects with VP (N = 7), PD (N = 21), and HCs (N = 58), the latter of which had cardiovascular risk factors but no neurological symptoms. The VP and PD groups were evaluated by a parkinsonism-focused motor exam and brief cognitive testing. We compared BG motor loop connectivity between groups and investigated for correlation between connectivity and clinical scores. To account for differences in fiber counts due to the different imaging scanners and protocols between cohorts, we used a BG motor loop proportion, which was the ratio of the BG motor loop fiber count over a control loop, the visual processing pathway. We used Kruskal-Wallis rank sum test with post-hoc Dunn tests to assess imaging findings between subject groups, and Pearson’s correlation to look for correlation between clinical scores and fiber counts. RESULTS/ANTICIPATED RESULTS: The whole brain connectome showed the fewest number of fibers in VP, followed by PD, and then HC (p<0.0001). The BG motor loop proportion fiber count of the BG motor loop was lower in the VP group, compared to the PD and HC cohorts (p = 0.031). In the VP group, the whole brain connectome fiber count correlated with a gait and balance subscore of the Movement Disorders Society - Unified Parkinson Disease Rating Scale (R = −0.87, p = 0.01). DISCUSSION/SIGNIFICANCE OF IMPACT: This study indicates that VP is associated with decreased structural connectivity, with a disproportionate degree of loss in the BG motor circuitry. While the etiology for this susceptibility to injury and preferential damage to BG remains to be defined, these findings can provide an important starting point for a biological understanding of VP, and a potential future marker for diagnosis and tracking disease progression.

OBJECTIVES/GOALS: The objective of the study was to determine whether CLP altered genes associated with mitochondrial function in the diaphragm. METHODS/STUDY POPULATION: A rodent cecal-ligation and puncture (CLP) model used to mimic sepsis-induced critical illness. The CLP model involved ligation of 50% of the cecum below the ileocecal valve in adult C57BL6 mice, followed by needle puncture of the cecum resulting in mid-grade sepsis. Mice survived for 48 hours or more, following injury. Diaphragm and limb muscles were harvested 24 hours following CLP (N = 6) and following a sham CLP procedure (N = 6). RESULTS/ANTICIPATED RESULTS: Gene expression of mitochondrial related genes (mef2c, myh1, pgc1-α), were significantly decreased in the diaphragm of CLP injured animals when compared to controls. In addition, ubiquitin ligases, genes associated with skeletal muscle atrophy murf1 and atrogin were increased in the diaphragm 24 hours after injury (p< 0.01). DISCUSSION/SIGNIFICANCE OF IMPACT: Our results indicate that sepsis-induced critical illness significantly impacts the expression of genes implicated in mitochondrial homeostasis and atrophy. Ongoing studies will identify whether CLP injury decreases skeletal muscle mitochondrial function.

OBJECTIVES/GOALS: We previously reported that genetic polymorphisms in the beta-catenin gene (CTNNB) are associated with the development of Clostridiodes difficile colitis during autologous stem cell transplantation (https://www-ncbi-nlm-nih-gov.proxy.libraries.uc.edu/pubmed/29594489). To biological validate these findings, we sought to evaluate the development of chemotherapy-associated Clostridiodes difficile infections by assessing the effect of C.difficile toxin B (TcdB) and of using melphalan in beta-catenin protein expression in Caco2 cells. METHODS/STUDY POPULATION: To determine the effect of melphalan and/or C.difficile toxin B on expression of Beta-catenin from human gut epithelial cells:

• Adenocarcinoma cells (Caco-2) cells were seeded and allowed to grow into monolayers

• Monolayers were treated with PBS, TcdB, melphalan and/or TcdB + melphalan for 24 hours and then washed with PBS

• Immunofluorescence was measured on the monolayers to visualize three markers -DAPI-Nuclear Stain (blue),Actin-ccytoskeletal stain (red), B-Catenin (green)

• Analysis of images with ImageJ (NIH). Statistical analysis of the effect of TcdB and/or melphalan on β-catenin protein levels was determined by One-way ANOVA

OBJECTIVES/GOALS: Variants in voltage-gated sodium channels (VGSC) are a common cause of severe early onset epilepsy. Changes in CSF neurotransmitters (NT) were identified in 2 cases of VGSC-related epilepsy. Here we investigate NT changes in patients and a novel mouse model of VGSC-related epilepsy. METHODS/STUDY POPULATION: We conducted a single site IRB approved retrospective chart review of patients with VGSC-related epilepsy who underwent CSF NT testing for diagnostic purposes. In parallel, we examined NT levels from the brains of wild-type (WT) and a novel VGSC-related epilepsy mouse model after obtaining IACUC approval. We rapidly isolated forebrain, cortex, striatum, and brainstem from 5-6 animals per sex and genotype. A combination of HPLC with electrochemical detection and mass spectrometry were used to quantify NT levels from brain samples. RESULTS/ANTICIPATED RESULTS: We identified 10 patients with VGSC-related epilepsy who received CSF NT testing. Two of these patients had abnormal NT results including changes to dopamine (DA) or serotonin (5-HT) metabolites. We analyzed NT levels from four brain regions from male and female WT and VGSC-related epilepsy mice. We anticipate that most of the NT levels will be similar to WT, however subtle changes in the DA or 5-HT metabolites may be seen in VGSC-related epilepsy. DISCUSSION/SIGNIFICANCE OF IMPACT: Patients with VGSC-related epilepsy often have autism spectrum disorder, sleep, and movement disorders. Understanding the role of aberrant NT levels in VGSC-related epilepsy may provide additional therapeutic targets that address common neuropsychological comorbidities as well as seizures.

OBJECTIVES/GOALS: Juvenile idiopathic arthritis (JIA) is the most common childhood rheumatologic disease childhood and a cause of pain and potential disability. JIA has a strong genetic component and no known cure. The goal of this study is to evaluate allele-dependent effects of a novel JIA risk variant at 1q24.3. METHODS/STUDY POPULATION: JIA patients meeting criteria for the two most common disease subtypes (oligoarticular and RF neg polyarthritis) were genotyped using the Immunochip, an Illumina array with dense coverage of the HLA region and 186 other loci previously reported in autoimmune diseases. Phase I association findings (Hinks, 2013) and Phase II analysis (unpublished) of an expanded cohort (4,271 JIA and 14,390 controls) identified new risk loci, including rs78037977 at 1q24.3. We prioritized rs78037977 and predicted possible impacted mechanisms based on Bayesian predictions of attributable risk, the surrounding chromatin landscape, and transcription factor binding data. A luciferase reporter assay was used to assess allele-dependent enhancer activity. RESULTS/ANTICIPATED RESULTS: rs78037977 is located between FASLG and TNFSF18 at chromosome 1q24.3 is associated with JIA (p = 6.3x10−09), and explains 94% of the posterior probability at this locus; no other SNPs in linkage disequilibrium (r2>0.6). The chromatin landscape around rs78037977 contains H3K4Me1 and H3K27Ac marks, which are indicative of enhancer activity. Further, >160 transcription factors have chromatin immunoprecipitation followed by sequencing (ChIP-seq) peaks overlapping rs78037977 in various cellular contexts. In luciferase reporter assays, the region around rs78037977 containing the reference A allele had ~2-fold increased enhancer activity compared to the non-reference allele. DISCUSSION/SIGNIFICANCE OF IMPACT: This work provides in vitro evidence to support allele-dependent enhancer activity of a novel JIA-risk variant at 1q24.3. Our ongoing work investigates the effect of the DNA-containing region of rs78037977 on gene expression and differential transcription factor binding at rs78037977.