OBJECTIVES/GOALS: Patient beliefs and goals can facilitate discussion of recovery expectations, patient-provider collaboration and maximization of goal achievement. In this study, we sought to address an evidence gap and examine the association of preoperative self-assessment of goals with preoperative and 6-week knee function and gait speed among total knee arthroplasty (TKA) patients. METHODS/STUDY POPULATION: We conducted a secondary analysis of data from the VERITAS randomized, controlled trial conducted from 11/2016-03/2018 that included adults age ≥ 18 years with scheduled and completed unilateral TKA followed by post-surgical physical therapy. Patients rated their ability to perform various activities of daily living goals scaled from 0 (unable to perform) to 10 (full performance). Patients were categorized by pre-surgical (baseline) goal rating: low = 0-2, intermediate = 3-4, and high = 5-10. Outcomes including gait speed and the KOOS were assessed within 10 days prior to surgery and 6-weeks post-surgery. Descriptive statistics and outcomes were compared for patients by preoperative goal rating using Chi-square or Fisher’s exact tests and ANOVA or Kruskal-Wallis tests as appropriate. RESULTS/ANTICIPATED RESULTS: Of 288 patients (mean age 65±8; 62.5% women; 82% white), 102 had a low goal rating (GR), 86 intermediate, and 99 high. Patients with low GR preoperatively generally had lower baseline mean scores than intermediate and high GR patients, respectively, on the KOOS (33.9/35.6/39.8; p<0.001) and lower gait speed (m/s) compared to intermediate and high GR patients at baseline (0.9/1.1/1.0; p = 0.009). The low, intermediate, and high GR groups, respectively, showed no difference across mean KOOS scores (61.0/61.2/61.9; p = 0.63) or gait speed (m/s) (1.0/1.0/1.0, p = 0.33) at 6 weeks postoperative. DISCUSSION/SIGNIFICANCE OF IMPACT: In this study, adults who perceived greater difficulty with a pre-selected activity goal, exhibited lower function prior to TKA but showed no differences in function 6-weeks after surgery. Follow-up studies will describe the association between goal-setting preoperatively and patient goal attainment and satisfaction following surgery.

ABSTRACT IMPACT: Our research highlights the need for both parental and clinical support to promote PA engagement among higher risk youth with comorbid asthma and obesity; these findings will inform research and clinical efforts in the youth development, prevention science, and clinical psychology fields. OBJECTIVES/GOALS: Asthma incidence doubles in youth with obesity. Physical activity (PA) is beneficial for asthma management; however, parental influence on PA levels among youth with asthma and obesity is poorly understood. This study examines the association of parents and PA among youth with asthma and/or obesity, accounting for risk and protective factors. METHODS/STUDY POPULATION: Data from 5th, 8th, 9th, and 11th-graders were obtained from the 2019 Minnesota Student Survey (N=96,820). Linear regressions examined the impact of parent connectedness on PA across 4 groups (neither asthma nor obesity [OB], asthma only, OB only, comorbid asthma/OB). The p-value for significance was set at p<.001. For PA, youth reported how many days they were physically active (≥60 min/day) in the last week. Two items assessing youth perception of parent care and ability to talk to parents about their problems were used to measure parent connectedness. BMI was calculated using self-report height/weight, age, and gender. Control variables included age, race/ethnicity, and free/reduced lunch eligibility. Models 2-4 retained parent connectedness variables and added risk and protective factors. RESULTS/ANTICIPATED RESULTS: In Model 1, both parent variables significantly predicted PA for each risk group (β ranges: parent care=.07-.09; parent talk=.04-.05, p<.001), except for the asthma/OB group (parent talk: p>.001). Models 2 and 3 added risk factors. Depression was the most salient risk factor, particularly for the highest risk group (asthma/OB; β =-.13, p<.001). Safe neighborhood was positively associated with PA for all groups (βs= .05, p<.001) except the asthma/OB group (p>.001). In Model 4, extracurricular activity involvement (protective factor) was positively associated with PA across all groups (β ranges=: .07-.11, p<.001), and depression remained significant across all groups (β ranges=-.11 to -.14, p<.001). For models 2-4, only parent care remained significant for the neither asthma nor OB group (β =.04, p<.001). DISCUSSION/SIGNIFICANCE OF FINDINGS: Results demonstrate that although parent care is an important protective factor for youth PA engagement, it is less impactful when additional risk factors (e.g., depression) are present, particularly among the highest risk group (comorbid asthma/OB). Thus, clinical support is needed in addition to parent support among higher risk youth.

OBJECTIVES/SPECIFIC AIMS: Given the poor prognosis of HCC and its increasing incidence worldwide, identifying biomarkers of HCC has been an active area of research. While biomarkers are being identified at a rapid pace, many are still in early phases of clinical study and very few have proven clinical utility. The objective of this study is to identify novel biomarkers of HCC and evaluate their clinical utility as predictors of disease development and prognosis with specific emphasis on disease recurrence after liver transplantation. Biomarkers will be identified through GWAS, as well as through analysis of qualitative and quantitative liver traits by magnetic resonance imaging (MRI). These novel biomarkers will then by implemented into risk prediction models aimed to assess an individual’s risk for development of HCC and stratify their level of risk according to predicted disease prognosis. METHODS/STUDY POPULATION: This will be a case-control study, analyzing data from previously created biorepositories from four cohorts of recipients across multiple centers which have undergone liver transplant. First, a GWAS will be performed to identify genetic variant(s). Second, pre-transplant MRI’s will be evaluated using CAVASS software to assess liver quantitative and qualitative traits, including visceral adiposity. Lastly, these findings will be implemented into risk stratification models to assess each individual’s level of risk for development of HCC and for recurrence of HCC after transplant. RESULTS/ANTICIPATED RESULTS: We hypothesize that genetic variant(s) are associated with positive HCV status and the development of HCC. Additionally, we hypothesize that increased visceral adiposity measured by MRI will have an association with recurrence of HCC after transplant. Lastly, we hypothesize that possession of these aforementioned features will be associated with an increased risk of HCC development and recurrence after transplant. DISCUSSION/SIGNIFICANCE OF IMPACT: As more is learned about the nature and reliability of these biomarkers, their potential clinical applications will be revealed. Ideally these proposed risk score models will ultimately be used by clinicians to provide personalized disease management while optimizing the allocation of health care resources. For instance, this may lead to changes in the MRI screening frequency of patients considered to be at high risk for HCC. The ability to diagnose patients early and provide personalized therapies may ultimately result in fewer disease related mortalities in the future.

OBJECTIVES/GOALS: The long-term goal of our lab is to develop clinical and intra-operative methods to aid in assessment of compressive and traumatic peripheral neuropathies. The overall objective of this project is toidentify the potential of serum neurofilament light chain as a diagnostic biomarker for nerve injury. METHODS/STUDY POPULATION: The objective of this prospective study is to obtain data on serum NfL levels in patients with cubital tunnel syndrome and traumatic nerve injuries. Serum NfL from patients with cubital tunnel and traumatic nerve injuries will be compared to serum NfL of asymptomatic, sex and aged matched controls. Pre-operative and post-operative serum levels will be measured and compared topatient’s pre-operative physical exam findings, motor and sensory function testing, electrodiagnostic studies, ultrasound, presence of intraneural vascularity, and post-operative patient reported outcome measures for cubital tunnel patients. For patients with traumatic nerve injury, acute phase and a subsequent serum NfL measurement will be used to assess temporal changes in NfL. RESULTS/ANTICIPATED RESULTS: The central hypothesis of this study is that symptomatic compression of the ulnar nerve or traumatic injury to the brachial plexus leading to axonotmesis will result in measurable increases in serum NfL proportional to the degree of nerve injury. This hypothesis has been formulated based on clinical experience and published studies demonstrating increased expression of serum NfL levels with axonal injury secondary to varying forms of peripheral neuropathy. DISCUSSION/SIGNIFICANCE: The significance of this project is that characterization of the relationship of clinical symptoms, non-invasive imaging and expression of NfL will lead to better diagnostic and prognostic algorithms in the treatment of compressive and traumatic peripheral neuropathies.

OBJECTIVES/GOALS: Viral acute rhinosinusitis (ARS), a.k.a, the common cold, affects millions every year. The symptoms caused by viral ARS dramatically affect the general well-being and functional levels of patients, causing work and school absence, and antibiotic abuse. In this study, we examined the therapeutic potential of topical adenosine in viral ARS METHODS/STUDY POPULATION: Rhinosinusitis was induced in WT and adenosine receptor (AR) knockout mice by respiratory syncytial virus (RSV) infection in the upper airways. Mice were subjected to adenosine or vehicle control within the sinuses. Adenosine receptor expression, inflammatory cytokine expression, and histologic mucus and inflammation score was assessed. The effect of endogenous adenosine accumulation within the sino-nasal tract was assessed in adenosine deaminase knockout (ADA-/-) mice. RESULTS/ANTICIPATED RESULTS: Topical administration of adenosine significantly inhibited the expression of pro-inflammatory cytokines, mucus production, and cell damage in the nose of mice with viral ARS, without prolonging virus clearance. This inhibitory effect was primarily mediated by the A2A adenosine receptor (AR). We also examined and compared the expression of ARs in the nasal tissue, trachea, and lungs. The nasal tissue exhibited the lowest baseline expression of ARs as compared to the lung and trachea which was further downregulated following adenosine treatment. Additionally, accumulation of endogenous adenosine in ADA-/- mice showed no signs of inflammation within the nasal tissue. Together, we demonstrated that topical adenosine effectively decreased inflammation and mucus production in a mouse model of viral ARS. DISCUSSION/SIGNIFICANCE: Previously, we found that topical adenosine dramatically enhances mucociliary clearance in the nose and sinuses. In this study, we found that nasal topical adenosine effectively decreased inflammation and mucus production in viral ARS. Our data suggest that nasal topical adenosine is an effective topical therapeutic option for viral ARS.

OBJECTIVES/GOALS: Diabetes mellitus is a common metabolic disease. Uncontrolled diabetes can lead to complications. The objective of this study is to examine the racial/ethnic variation in the relation between diabetes control and healthy eating, food security, exercise, and access to health care. METHODS/STUDY POPULATION: We analyzed data related to diabetes control, demographics, insurance, healthy eating, food security and physical activity for 949 diabetics from the National Health and Nutrition Examination Survey (NHANES). The population examined was adults with diabetes mellitus. Diabetes control was classified as fair control [HbA1c 7-<8 and fasting glucose <126 mg/dL], good control [HbA1c<7 and fasting glucose <110 mg/dL] or uncontrolled [HbA1c8 and fasting glucose <110 mg/dL]. We used multinomial logistic regression controlling for confounders to analyze the data overall and for each racial/ethnic group and report adjusted odds ratios (AOR) and 95% confidence limits (CL). RESULTS/ANTICIPATED RESULTS: Of the 949 diabetics, 14.7% were Blacks, 15.9% were Hispanics, 11.0% had fair control, 61.0% had good control, 14.2% were uninsured, 18.1% had low/very low food security, and 39.7% were inactive. Overall, uninsured subjects had a lower chance of fair diabetes control (AOR = 0.2, 95% CL = 0.1-0.9, p = 0.04), but this relationship was significant only for Hispanics and Blacks (p<0.05). Whites with low/very low food security were less likely to have fair control (AOR = 0.2, 95% CI = 0.001-0.9, p = 0.04). Blacks who were inactive, and insufficiently active Hispanics had a lower chance of good/excellent diabetes control (AOR = 0.5, 95% CI = 0.2-0.9, p = 0.03; AOR = 0.3, 95% CI = 0.1-0.7, p = 0.007 respectively). DISCUSSION/SIGNIFICANCE OF IMPACT: The results show the importance of insurance coverage, food security and physical activity in diabetes control among different racial/ethnic groups. They indicate a need for affordable health care and for culturally-relevant interventions that include physical activity and food security.

In addition to facing numerous healthcare disparities, rural America is chronically underrepresented in clinical research. This gap was made more evident during the COVID-19 pandemic. St Lawrence Health, located in rural Upstate New York, established its Clinical and Rural Health Research Department in 2015 to help close this gap. The research department then launched the DISRUPTS (Developing InfraStructure for Research to Utilize Patient-centered Techniques at St Lawrence Health System) program to build the infrastructure to conduct Patient-Centered Outcomes Research (PCOR). Together with a diverse committee, the team used proven methods and frameworks to develop a model for engagement, content creation, and education delivery that was successfully used to create educational programs on PCOR and COVID-19. The resulting DISRUPTS webinars had a combined total of over 450 live attendees and over 1,110 views on recordings. Furthermore, nearly one-third of those who participated in the COVID-19 vaccines webinar indicated they were more likely to receive a COVID-19 vaccine after taking part. DISRUPTS can serve as an important model for other rural communities that aim to increase access to and engagement in PCOR, and which hope to improve outreach and education efforts in their communities.

OBJECTIVES/SPECIFIC AIMS: 1) Describe strategies pediatric providers perceive improve chlamydia screening of sexually active female adolescents (SA), and 2) describe barriers to regular screening of SA for chlamydia METHODS/STUDY POPULATION: Using qualitative methods, 14 general pediatric providers across 7 clinical sites in Vermont were interviewed to ascertain best practices and remaining challenges. Semi-structured interviews lasting 30-45 minutes were audiotaped and transcribed. Chlamydia screening rates provided by BCBS-VT were used to categorize participant responses across three performance tiers, data were coded, and themes identified within these tiers. RESULTS/ANTICIPATED RESULTS: Facilitators: When asked to describe facilitators of chlamydia screening, providers in the top tier of chlamydia screening emphasized the importance of adequate insurance to cover the cost of testing. Providers in the middle performance tier cited use of pre-visit questionnaires, and those in the bottom performance tier identified no best practices. Other strategies included improving physician confidence and awareness, establishing practice- and individual-level routines, and providing strong leadership and communication of local screening rates. Barriers: Across the 3 performance tiers, the most common challenges to consistent chlamydia screening were threats to patient confidentiality, cost of the screening test, and requirement for patient disclosure of sexual activity. Less commonly, providers were concerned that adolescent patients were not reliable to obtain screens off-site, or fill treatment prescriptions without the help of a parent. DISCUSSION/SIGNIFICANCE OF IMPACT: The need for systematic, confidential, and inexpensive means for screening SA for chlamydia was highlighted in both the best practices and challenges described by providers of pediatric care in the suburban practice setting. Policy and practice interventions may target these needs to improve the reproductive health of female adolescents.

ABSTRACT IMPACT: For community engagement to be impactful and reduce health inequity, it needs to address timely needs in the community, including COVID-19 impacts. Here, we describe how pre- and post-COVID-19 food insecurity worsened mental health among community members served by HealthStreet University of Florida community engagement program. OBJECTIVES/GOALS: COVID-19 impacts the economic vitality and the mental health of communities; research and engagement activities must consider the context in which we are practicing and the needs of our community members. METHODS/STUDY POPULATION: HealthStreet, the University of Florida community engagement program, sends Community Health Workers (CHWs) where people congregate to assess social determinants of health and medical histories, used to make referrals to services and research opportunities. CHWs conducted follow-up COVID-19 assessments measuring perceived stress, loneliness, depression, anxiety, binge drinking, and opioid use, as well as high blood pressure and food insecurity. Here, we consider mental health outcomes among 1,300 adults who reported being food insecure either at some time in the past 12 months at baseline, or at the COVID-19 follow-up assessment, and completed both. Chi-Square Test was used to determine p-values. RESULTS/ANTICIPATED RESULTS: Overall, at the COVID-19 follow-up assessment, 37.1% (of 1,300) were still food insecure during COVID-19 (same), 20.3% (had become food insecure during COVID-19 (worse) and 42.6% were no longer food insecure (better). Those who were no longer food insecure were more likely to report less stress, while those still food insecure were more likely to report the highest stress and loneliness (p<0.0001), while the worse off group was in the middle. Those who stayed food insecure were most likely to report depression and anxiety, and also high blood pressure and using opioids (p<.05) compared to those getting worse or better. Binge drinking behavior was not significantly different across groups. DISCUSSION/SIGNIFICANCE OF FINDINGS: Community engagement activities across CTSIs must be sensitive to the needs of their communities. HealthStreet findings show that new and continuing food insecurity negatively influence mental health problems, pointing to the need for engagement to address multiple problems.

OBJECTIVES/SPECIFIC AIMS: There is currently a gap in the literature regarding the relationship between acute cellular rejection and frailty in LT patients. We aimed to evaluate the association between frailty and acute cellular rejection in LT patients. METHODS/STUDY POPULATION: Included were LT recipients from 2014-16 at a single center who had a frailty assessment prior to LT using the Liver Frailty index consisting of grip strength, chair stands, and balance. Frailty was defined by a Liver Frailty Index > 4.5. Data on acute cellular rejection at 3 months (primary outcome) and immunosuppression regimens were collected from medical chart review. Univariable and multivariable logistic regression assessed the associations between frailty and acute cellular rejection. RESULTS/ANTICIPATED RESULTS: A total of 241 LT recipients were included. Of these, 37% were female, 55% had Hepatitis C, and the median (IQR) age was 60 (54-65); 46 (19%) were classified as frail. 98% of patients were on a combination of mycophenolate, corticosteroids and tacrolimus on discharge compared to 80% by 3 months. Within the first 3 months post-LT, 7 (15%) of frail patients versus 10 (5%) (p = 0.02) of non-frail patients experienced acute cellular rejection. In univariable logistic regression, frailty was associated with a 3.3 times higher odds of acute cellular rejection at 3 months (95%CI 1.19, 9.26, p = 0.02); age (OR 0.91), Black race (OR 3.2), autoimmune disease (OR 2.3), and diabetes (OR 0.3) were also associated with acute cellular rejection at 3 months with a p-value<0.20. In a multivariate analysis, after adjusting for age, frailty remained significantly associated with rejection (OR 3.06, 95%CI 1.04, 9.01, p = 0.043). There were no significant differences in immunosuppression regimens or rates of mycophenolate dose reduction in the first 3 months between frail and non-frail patients. DISCUSSION/SIGNIFICANCE OF IMPACT: Frailty is associated with an increased rate of acute cellular rejection within 3 months post-LT, despite similar immunosuppression regimens and doses. Future studies should evaluate whether frailty should be considered in the management of immunosuppression in the early post-transplant period.

OBJECTIVES/GOALS: COPD is a progressive airways disease that results in death or disability. There is poor uptake of clinical guidelines (CPG) to manage COPD and studies to bridge this implementation gap have shown inconsistent results. Using implementation science principles we aim to understand COPD-CPG implementation determinants from providers’perspective. METHODS/STUDY POPULATION: The study is being conducted in ten VA Primary Care Clinics. Guided by the Consolidated Framework for Implementation Research (CFIR), a conceptual framework developed to guide systematic assessment of multilevel implementation contexts, we are using semi-structured guides to conduct key informant qualitative interviews (physicians, physician extenders and nurses), to support a formative evaluation. CFIR domains relevant to the study were determined by a multidisciplinary team. Informants are identified through online outreach and voluntary participation. Sampling adequacy will be assessed by achievement of code saturation. A qualitative template analysis will be used to summarize the barriers and facilitators of each component of COPD-CPG organized by CFIR-domain. RESULTS/ANTICIPATED RESULTS: We anticipate a list of modifiable and non-modifiable contextual, recipient (provider and patient), and COPD CPG content (innovation) barriers to implementation. Many settings do not have critical elements of these CPG, such as a standardized inhaler education/assessment pathway, patient education material, or pulmonary rehabilitation referral pathway. Existing literature indicate reasons behind the insufficient uptake of COPD CPG include low familiarity with guidelines, perception of minimal value of guidelines by physicians, and time constraints; we will present contextual, recipient and innovation determinants specific to our setting. DISCUSSION/SIGNIFICANCE: This comprehensive assessment of barriers and facilitators to COPD-CPG will inform tool development and implementation strategies identification to improve COPD CPG uptake. COPD is the most common veteran lung disease. Improvement in COPD care has enormous potential for benefit for local veterans, as well as potential for wider dissemination.

ABSTRACT IMPACT: Better understanding of the factors impacting disease severity and immunological response of MS patients on disease modifying therapy will enable better recommendations for vaccination options and risk mitigation strategies OBJECTIVES/GOALS: The Coronavirus Disease 2019 (COVID-19) and global health crisis has raised health concerns for patients with multiple sclerosis (MS). We aim to study the clinical characteristics, immunological laboratory data, and immunoglobulin response in patients with MS and COVID-19, to identify factors impacting disease severity and immune response. METHODS/STUDY POPULATION: Database search was done using DataDirect to search for MS patients who had tested positive for COVID-19 at the University of Michigan hospital. Patients with a positive nasopharyngeal swab polymerase chain reaction (PCR) for COVID-19 between March 1 and September 2020 were included. The primary outcome was the immunological laboratory data and immunoglobulin levels and the secondary outcome was their disease severity. We collected demographics, neurological history, MS treatment, Expanded Disability Scale Score (EDSS), comorbidities, and COVID-19 characteristics. A 7-point ordinal scale previously used to assess disease severity was used. Univariate and multivariate analyses will be performed to assess relationships between the collected variables. RESULTS/ANTICIPATED RESULTS: A total of 17 patients, mean age 53 (SD 11.6) years, mean disease duration, 6.2(SD 4.1) years were analyzed. 41% of patients had relapsing remitting multiple sclerosis, 17% had primary progressive MS. (88%) patients were on Disease Modifying Therapy (DMT) at the time of COVID-19 diagnosis. 2 patients died from COVID-19 complications. There was a higher proportion of patients with higher disease severity receiving Ocrelizumab. Only one patient showed positive IgG to SARS-CoV-2 after the resolution of infection. CBC with differential was obtained and a longitudinal follow-up of labs will be done. Regression analysis will be done to check the association between the use of DMT, immunological response, and COVID disease severity in them. The impact of COVID-19 on MS relapse, EDSS, and MRI activities will also be studied. DISCUSSION/SIGNIFICANCE OF FINDINGS: Recommendations to continue current DMT have been made, however, the immune response has not been correlated with the individual’s risk profile. Certain therapies may interfere with mounting a protective immune response of COVID-19 and this knowledge is crucial when advising patients regarding the choice of vaccine and risk mitigation strategies.

OBJECTIVES/GOALS: Lung transplant (LTx) candidates benefit from use of non-ideal donor organs. Each organ procurement organization (OPO) defines “acceptable” donor organs introducing unmeasured variation in donor pursuit. We characterized non-ideal donor pursuit among OPOs to identify drivers of risk aversion in LTx. METHODS/STUDY POPULATION: We queried the UNOS registry for adult donors who donated ≥1 organ for transplantation from 12/2007-12/2018. Non-ideal donors were those with any of age>50, smoking history ≥20 pack-years, PaO2/FiO2 (P/F) ratio<350, donation after cardiac death (DCD) status, or CDC increased risk (IRD) status. Non-ideal donor pursuit rate was defined as the proportion of non-ideal donors at each OPO from whom consent for lung donation was requested with lower numbers indicating increased risk aversion. We estimated the correlation between non-ideal and overall donor pursuit using a Spearman correlation coefficient. Adjusted non-ideal donor pursuit rates were estimated using multivariable logistic regression. RESULTS/ANTICIPATED RESULTS: Overall, 18,333 deceased donors were included and classified as ideal or non-ideal. Among 58 OPOs, rates of non-ideal donor pursuit ranged from 0.24-1.00 Figure). Of 5 non-ideal characteristics, DCD and IRD status were associated with the most and least risk aversion, respectively. Non-ideal donor pursuit was strongly correlated with overall donor pursuit (r = 0.99). On adjusted analysis, older age (OR 0.15, 95% CI 0.13-0.16), smoking history (OR 0.38, 95% CI 0.34-0.44), low P/F ratio (OR 0.12, 95% CI 0.11-0.14), and DCD status (OR 0.04, 95% CI 0.03-0.04) were all independently associated with significant risk aversion, corresponding to decreased rates of donor pursuit. DISCUSSION/SIGNIFICANCE OF IMPACT: OPOs differ in their levels of risk aversion in LTx and risk aversion is not uniform across selected categories of non-ideal lung donor. Consideration of new OPO performance metrics that encourage the pursuit of non-ideal lung donors is warranted.

OBJECTIVES/GOALS: Dyslexia is a common Reading Disability (RD) affecting 7-12% of the population and is associated with less cortical thickness (CT) in bilateral brain regions. However, the interaction between RD and a bilingual experience on CT is unknown, even though bilingualism is also associated with altered CT. METHODS/STUDY POPULATION: We studied 48 Bilinguals assigned to the Typical Reader group based on Oral Reading Recognition Test (ORRT) scores above 90 (avg=107 ± 14), 47 Bilinguals assigned to the RD group based on ORRT scores below 85 (avg=77 ± 5), 45 English Monolingual Typical Readers with ORRT scores above 90 (avg=102 ± 13) and 47 Monolinguals with RD based on ORRT scores below 85 (avg=78 ± 5). Participants (all from the Adolescent Brain & Cognitive Development Study) were 11.9 ± 0.7 years of age and the 4 groups were matched for sex, self-ratings of English, nonverbal reasoning, and combined household income. Structural magnetic resonance images were analyzed using CAT12 and all four groups were entered into a factorial analysis. RESULTS/ANTICIPATED RESULTS: Surprisingly, the main effect of Reading Ability did not reveal any regions where RD manifested less CT than Controls (raising the possibility that the findings from the only two prior reports were due to small samples). The main effect of Language Background revealed less CT in bilinguals in bilateral perisylvian regions (inferior frontal gyri, superior temporal gyri, and left Heschl's gyrus) consistent with prior reports. There was no interaction of Reading Ability by Language Background. Taken together, we found no differences in CT in those with RD relative to Typical readers and no evidence that the dual language experience affected this result in any way. DISCUSSION/SIGNIFICANCE: The lack of interaction between Reading Ability and Language Background indicates that a dual-language experience does not affect CT differently in those with RD and reduces concerns that RD in those who are bilingual needs to be given separate consideration in studies of CT neuroanatomy.

OBJECTIVES/GOALS: Assess molecular and cellular mechanisms of allograft loss in kidney biopsies using digital spatial profiling and clinical outcomes data. METHODS/STUDY POPULATION: Patients with chronic allograft dysfunction (CGD), enrolled in the Deterioration of Kidney Allograft Function (DeKAF) study, with or without eventual allograft loss, were included. CGD was defined as a >25% increase in creatinine over 3 months relative to a baseline. Kidney biopsy tissue was assessed by Nanostring GeoMX digital spatial profiling (DSP) after staining with anti-pan-cytokeratin, anti-CD45, anti-CD68, Syto-13, to identify specific cell populations, and Nanostring’s Whole Transcriptome Atlas (WTA), to quantify the distribution of transcripts across the biopsy. Up to 14 regions of interest (ROIs) were selected, with or without glomerulus. CIBERSORT was used to perform cell deconvolution. Clinical and outcomes data were from the DeKAF study and United States Renal Data System. RESULTS/ANTICIPATED RESULTS: Macrophage (M1) cell population abundance was significantly different in ROIs with glomerulus between graft loss and no graft loss. Principle component analysis of differentially expressed genes resulted in transcriptomes in ROIs that cluster together by clinical outcome of graft loss or no graft loss. There were 203 DEGs in ROIs with glomerulus that were different by graft loss or no graft loss. By pathway analysis, these 203 DEGS were enriched in the T-cell activation, integrin signaling and inflammation pathways. DISCUSSION/SIGNIFICANCE: DSP of kidney allograft biopsies allows for the identification and quantification of specific cell types, such as macrophages and molecular transcripts as potential drug targets. This data can be used to understand mechanisms of kidney allograft loss and may lead to improved immune suppression in kidney transplant recipients.

OBJECTIVES/GOALS: Tubulointerstitial damage in lupus nephritis (LN) is a strong predictor of progression to chronic kidney disease and end stage renal disease (ESRD). While complement activation mediates glomerular injury, the role of complement in renal tubular damage has not been evaluated. We investigated the association between complement activation and tubulointerstitial fibrosis. METHODS/STUDY POPULATION: Patients with LN were selected randomly between July 2014 - July 2016. Chromogenic immunohistochemistry was performed on formalin-fixed, paraffin-embedded, 4-µm human renal biopsy sections using unconjugated, murine anti-human Complement C9 (Hycult Biotech, clone X197) as a marker of the terminal complement activation. Positive control is C3 glomerulopathy and negative control is normal kidney. Tubular basement membrane C9 staining intensity were analyzed on semiquantitative scale 0 to 3 by a renal pathologist. Interstitial fibrosis/tubular atrophy were categorized into low (0–10%), medium (11–20%), or high (≥21%). Clinical parameters were assessed at time of biopsy and 6 months post biopsy. Bivariate associations were assessed between presence of tubular C9 (C9+) and other covariates. RESULTS/ANTICIPATED RESULTS: Renal biopsies from 30 LN studied, 23 (77%) of which had proliferative LN. There were 24 (80%) women, mean (SD) age 33 (12) years. Positive tubular C9 staining was observed in 7/30 (23%) biopsies. At time of renal biopsy, C9+ patients had significantly higher urine protein, compared to C9- patients: median (IQR) 6.2g (3.3-13.1) vs. 2.4g (1.3-4.6), p<0.01. The differences persisted at 6 months after induction therapy: 1.08g (1.0-8.3) in C9+ vs. 0.68g (0.2-2.1) in C9- patients, p = 0.06. There was no significant difference in creatinine at renal biopsy between the two groups. Tubular C9 deposition was associated with interstitial fibrosis: 49% had severe interstitial fibrosis vs. none in the C9- group, p = <0.01. Higher proportion of C9+ patients had moderate NIH Chronicity index: 42.9% vs 8.7% in the C9- group, p = 0.07. DISCUSSION/SIGNIFICANCE OF IMPACT: Tubular C9 deposition is significantly associated with proteinuria, interstitial fibrosis and increased chronicity which predict progression to ESRD and high mortality. This finding suggests that complement activation in the tubules may be linked to proteinuria and contribute to mechanism in tubulointerstitial damage in LN.

OBJECTIVES/GOALS: Untargeted metabolomics platforms are powerful biomarker discovery tools. However, the absence of uniform study design, data analysis, and reporting standards limits translation of this research into the clinical lab. The goal was to critically appraise existing untargeted metabolomics platforms that analyzed inborn errors of metabolism. METHODS/STUDY POPULATION: A search strategy was conducted in MEDLINE via PubMed from January 16, 2013, to January 16, 2023. The search strategy was limited to primary literature articles written in English that evaluated human subjects with inborn errors of metabolism (IEMs). Articles that performed targeted metabolomic analysis or analyzed non-human samples were excluded. Information on patient cohorts analyzed, sample types, and method design were extracted using a template. Categorical data are summarized as frequencies and percentages. RESULTS/ANTICIPATED RESULTS: A total of 96 distinct IEMs were evaluated by the different untargeted metabolomics methods included in this review. However, most IEMs (55/96, 57%) were evaluated by a single platform, in a single study, with a limited cohort size. Only one study validated their results using a separate, validation cohort. There was considerable diversity in the separation techniques and mass spectrometry instrumentation used by the studies to create their untargeted metabolomics methods. Slightly over half (59%) of the studies identified at least some of the metabolites detected in their samples with the highest level of confidence. Importantly, most of the included studies reported adherence to quality metrics, including use of quality control material (65%) and internal standards in their analysis (61%). DISCUSSION/SIGNIFICANCE: Future studies analyzing IEM patient samples with untargeted metabolomics platforms should progress beyond single-subject studies and evaluate the reproducibility of the research using a prospective, or validation cohort as well as confirm metabolite annotations with reference metabolites standards to generate clinically useful data.

OBJECTIVES/GOALS: Social isolation/loneliness is a public health crisis and one that is unlikely to be solved through pharmacology. Nonpharmacological approaches, such as dance, are needed. The objective of this study is to investigate the physiological correlates of dance-induced improvements in social connection. METHODS/STUDY POPULATION: Participants were randomly assigned to participate for 4 weeks (2 times per week, 90-minute sessions) in either 1) improvisational dance training (experimental group; n=7); or a 2) dance movie watching experience (control group; n=7). Before and after the intervention, using mobile brain-body imaging techniques, participants and their instructor had their brain (via electroencephalography) and body physiology (via photoplethysmography) recorded during a series of verbal and nonverbal interactive experiences. Participants were also video recorded via 4 surrounding cameras for later motion capture analysis. Neuropsychological assessments were also conducted before and after the intervention. RESULTS/ANTICIPATED RESULTS: We found that dance significantly increased social skills including empathy, interpersonal skills, emotional regulation, mindfulness, and attention. Additionally, we found that dance significantly increased interbrain synchrony during nonverbal experiences including theta (4-8 Hz), beta (12-35 Hz), and gamma (35-45 Hz) frequencies in the occipital lobe. Increases in interbrain synchrony were also positively correlated with increases in empathy. Additionally, intercardiac synchrony between the participant and instructor showed a significant correlation at post-intervention only. Future investigations will focus on the relationship between interbrain, intercardiac, and movement synchrony. DISCUSSION/SIGNIFICANCE: Our findings support the idea that dance increases interpersonal synchrony at the level of the brain, heart, and behavior. Understanding the neural and somatic mechanisms of social behaviors will help promote understanding and development of interventions for the critical problem of social isolation and loneliness.