OBJECTIVES/GOALS: This study aimed to characterize urinary exosomal miRNA content in African American adults with diabetic kidney disease. METHODS/STUDY POPULATION: Male and female participants between the ages of 18 and 65 were recruited from the Diabetes Treatment Center and the Nephrology Clinic at the Howard University Hospital. Exosomes were isolated from cleared urine of healthy controls (n=3), type 2 diabetics (n=3), and participants with chronic kidney disease (n=3). The purity and size of isolated microparticles was evaluated using NanoSight technology (30nm to 120nm size range) and western blot analysis for exosome-specific markers (TSG101 and CD81) RESULTS/ANTICIPATED RESULTS: Expression of 5 selected microRNAs, miR-4534, miR-320c, miR-451, miR-362-3p and miR-877-3p were evaluated by qRT-PCR. miR-4534 and miR-451 was increased between healthy controls and the type diabetic group. MiR-320c was increased in the CKD group, in comparison to healthy controls. Conversely, there was no difference in miR-877-5p between the three groups. DISCUSSION/SIGNIFICANCE: These findings will provide insight into the use of circulating miRNAs as early markers of DKD, ultimately creating more effective treatments and preventive measures.

Underrepresentation of people from racial and ethnic minoritized groups in clinical trials threatens external validity of clinical and translational science, diminishes uptake of innovations into practice, and restricts access to the potential benefits of participation. Despite efforts to increase diversity in clinical trials, children and adults from Latino backgrounds remain underrepresented. Quality improvement concepts, strategies, and tools demonstrate promise in enhancing recruitment and enrollment in clinical trials. To demonstrate this promise, we draw upon our team’s experience conducting a randomized clinical trial that tests three behavioral interventions designed to promote equity in language and social-emotional skill acquisition among Latino parent–infant dyads from under-resourced communities. The recruitment activities took place during the COVID-19 pandemic, which intensified the need for responsive strategies and procedures. We used the Model for Improvement to achieve our recruitment goals. Across study stages, we engaged strategies such as (1) intentional team formation, (2) participatory approaches to setting goals, monitoring achievement, selecting change strategies, and (3) small iterative tests that informed additional efforts. These strategies helped our team overcome several barriers. These strategies may help other researchers apply quality improvement tools to increase participation in clinical and translational research among people from minoritized groups.

OBJECTIVES/SPECIFIC AIMS: We developed a multilevel hierarchical statistical model which describes the association of prophylactic interventions to patient PONV risk, and provides an intuitive summary for anesthesiologists to understand how well they are adhering to PONV guidelines. METHODS/STUDY POPULATION: Accepted PONV risk factors as well as preventative interventions to reduce the PONV risk, (e.g. total intravenous anesthesia or pharmacological prophylaxis) are retrieved from the electronic medical record (EMR). Risk is regressed against interventions. Fig 1, Panel A visualizes adherence for an individual provider by plotting anesthesia cases, with PONV risk in the x-axis and the number of interventions in the y-axis. Fig 1, Panel B shows a “Jitterplot”, jittering individual cases, which would otherwise plot onto the same coordinates (Panel A). The distribution of the number of interventions in each risk category is better summarized in Fig 1 Panel C by overlaying a violin plot onto the “Jitterplot”. Finally, a fitted regression line provides a summary measure for the individual provider’s risk-adjusted utilization of PONV prophylaxis in Fig 1, Panel D. The model can control for confounders and interactions, such as patient or procedure characteristics, such as supervision by attending physicians, institutional culture, and surgical procedure. RESULTS/ANTICIPATED RESULTS: Fig. 2, Panel A demonstrates good adherence. The provider responded to increased risk with additional interventions leading to a steep regression line. Less discriminate administration of prophylaxis is shown in Fig 2, Panel B. The graphical representation of our proposed measure of individual provider performance is intuitive, allowing us to compare adherence of two distinct groups of providers (light lines) and institutional averages (dark lines) as shown in Fig 2, Panel C. Controlling for known risk factors and potential confounders renders the assessment irrepudiable. The rigorous statistical approach allows for multi-level modeling and comparative effectiveness research, realistically evaluating process changes and interventions like CDS in the hierarchical structure of contemporary healthcare delivery. DISCUSSION/SIGNIFICANCE OF IMPACT: The strength of our novel measure of individual provider performance is its generalizability to other care settings, as well as the intuitive graphical representation of risk-adjusted individual performance. However, accuracy, precision and validity, sensitivity to system perturbations (like the implementation of CDS), and acceptance among providers remain to be evaluated. Fig 1. Risk-Adjusted Utilization of Antiemetic Prophylaxis Fig 2. Comparing Performance between Provider Groups

OBJECTIVES/GOALS: Little is known about the effect of hemodilution with crystalloid on blood coagulation in obstetric patients. The purpose of our study was to examine the impact of hemodilution on components of blood coagulation using rotational thromboelastometry (ROTEM®) in term parturients METHODS/STUDY POPULATION: This is a prospective, observational pilot study including 35 healthy, pregnant patients at term (≥37 weeks) without history of bleeding or clotting disorder or on medication affecting coagulation. Venous blood samples were collected from all patients and divided into specimen tubes to generate varying degrees of hemodilution with Plasma-Lyte (0%, 20%, 25%, 30%, 35%, 40%, 45%, 55%, 60%, 65%, 70%, 75%, 80%). Rotational thromboelastometry was then performed on samples to assess for coagulation changes. RESULTS/ANTICIPATED RESULTS: EXTEM (extrinsically activated assay) clotting time (CT) became prolonged at 65% hemodilution and above, and the median CT was in the coagulopathic range (>80 seconds) at a dilution of 80%. FIBTEM (extrinsically activated assay with platelet inhibitor, primarily measuring contribution of fibrinogen to coagulation) amplitude at 5 minutes (A5) began to diminish at 35% hemodilution, with the median A5 in the coagulopathic range (<12 mm) at 55% hemodilution. The area under the curve (AUC), a marker of clot strength, for EXTEM and FIBTEM consistently declined as hemodilution increased. Greater decreases in FIBTEM AUC were seen compared to EXTEM AUC, with the ratio of FIBTEM:EXTEM AUC at each dilution demonstrating a statistically significant difference from baseline. DISCUSSION/SIGNIFICANCE OF IMPACT: All thromboelastometry values demonstrated a hypocoagulable trend as hemodilution increased. However, the samples analyzed by the FIBTEM assay trended toward a coagulopathy at a lower degree of hemodilution compared to the EXTEM assay. As FIBTEM tests analyze the role of fibrinogen in hemostasis and EXTEM tests analyze the role of platelets, our findings suggest that platelets may be able to withstand higher degrees of hemodilution before impairing hemostasis compared to fibrinogen. These findings support the growing body of literature that in early stages of severe obstetric hemorrhage, the prioritization of fibrinogen replacement may be critical in preventing further coagulopathy. CONFLICT OF INTEREST DESCRIPTION: All authors have no conflicts of interest to report.

Objectives/Goals: To determine the heterogeneity in CD4:CD8 ratio in a well-characterized cohort of PWH and to investigate the predictors of intestinal CD4:CD8 ratio reconstitution (CD4:CD8>1) and its impact on systemic inflammation. Methods/Study Population: We enrolled 52 PWH on ART and with peripheral HIV-RNA Results/Anticipated Results: PWH had a lower CD4:CD8 ratio both in the peripheral blood [p1. This subset of PWH was more likely female (62% vs. 38%, p = 0.0158), diagnosed with HIV for a longer time [p = 0.0347] have longer duration of most recent viral suppression [p = 0.0365] higher CD4+ T cells at enrollment [p =  0.0262] and higher CD4+ T cell nadir. Multiple logistic regression showed that duration of HIV infection [OR 1.13 (95% C.I. 1.02–1.3)] and CD4 =  T cell nadir[OR 1.01 (95% C.I. 1.001–1.016)] were associated with colonic CD4:CD8 >1. Colonic CD4:CD8 ratio partially correlated with the peripheral blood CD4:CD8 ratio (r = 0.274, p = 0.068) and with the pro-inflammatory cytokines IL-20 (r =  -0.413, p = 0.036) and SLAMF-1 (r =  -0.329, p = 0.074). Discussion/Significance of Impact: In PWH, CD4:CD8 ratio

OBJECTIVES/GOALS: Chronic kidney disease (CKD) impacts 15% of US adults and African American (AA) persons are disproportionately affected with more than 3 times higher risk of kidney failure when compared to Caucasian persons. This study evaluated the physiological and metabolomic effects of increased fruits and vegetables (F&V) on cardio-renal risk factors. METHODS/STUDY POPULATION: This pilot trial used a prospective, 2-group, randomized study design to evaluate a F&V intervention (N=46), where participants received a prescribed amount of fresh, base-producing F&V compared to a wait-list control (WL) condition (N=45). All participants were African American adults (≥18 years), had self-reported hypertension, and had CKD (Stage 1-3) on screening spot-urine microalbumin test. Participants were measured at baseline and 6 weeks post-intervention. Clinical data (i.e., systolic and diastolic blood pressure, lipid panel, hemoglobin A1C, BMI [body mass index], and albumin to creatinine ratio) were collected. Targeted metabolomic quantitative analysis was performed followed by LC-MS/MS and FIA-MS/MS. Linear mixed models evaluated analyte expression and clinical data. RESULTS/ANTICIPATED RESULTS: AA participants (N=91) were aged 58 ± 10.2 years, 66% female, and 54% had incomes ≤$50,000. T-tests compared change scores (baseline to 6-weeks) between groups. The F&V group demonstrated a significant reduction in BMI of -4.7 ± 10.5 kg/m² compared to a 1.9 ± 8.3 kg/m² increase in the WL group, p<.01. Further, the F&V group demonstrated a reduction in total cholesterol of -15.4 ± 58.8 mg/dL compared to a 17.7 ± 68.8 mg/dL increase in the WL group, p<.05. Non-significant reductions in hemoglobin A1c were found in the F&V versus the WL group. Metabolomic analysis indicated significant variation with an increase of suggestive key biomarkers for worse CKD in the WL versus F&V groups at 6-weeks. DISCUSSION/SIGNIFICANCE: Consumption of only 2 cups of F&V via a community-based intervention reduced CVD risk factors in AA adults with CKD and HTN and resulted in molecular/biochemical changes which may improve long-term kidney health. Further investigation may lead to development of cost-effective dietary intervention models to improve CKD outcomes in AA persons.

OBJECTIVES/GOALS: The research objectives of this project are to elucidate the effects of Bruton’s tyrosine kinase inhibitors (BTKi) of varying target specificity on platelet function with regard to platelet aggregation, adhesion, spreading, and intracellular signaling as measured by kinase phosphorylation. METHODS/STUDY POPULATION: Blood from healthy volunteers was obtained and processed to obtain both washed platelets and platelet-rich plasma. The samples were then treated with one of the BTKi drugs or with vehicle (DMSO) at concentrations matching patient blood concentrations derived from clinical trials and pharmacokinetic studies. The incubated samples were then analyzed in an aggregometer using one of several agonists. Aggregation was stopped after five minutes with a perchloric acid-based lysis buffer. The samples were then analyzed by SDS-PAGE and immunoblotting to quantify BTK protein and BTK phosphorylation. Adhesion was assessed by incubating washed platelets treated with BTKi on microtiter wells coated with fibrinogen or collagen and quantifying adherent platelets by their endogenous acid phosphatase activity. RESULTS/ANTICIPATED RESULTS: We found that ibrutinib, zanubrutinib, and pirtobrutinib all completely inhibited collagen-induced platelet aggregation, whereas they did not inhibit aggregation induced by thrombin, ristocetin, arachidonic acid, or the PAR1 activator peptide SFLLRN (T6). Acalabrutinib inhibited collagen-induced platelet aggregation only at high concentrations (1-2 micromolar). At the lower concentration of 200 nanomolar, comparable to the concentration required for the other BTK inhibitors to completely inhibit platelet aggregation, acalabrutinib failed to inhibit aggregation but did inhibit auto-phosphorylation, indicating an impact on signaling. None of the BTKi drugs inhibited adhesion of platelets to collagen-coated surfaces. DISCUSSION/SIGNIFICANCE: Our data show the inhibitory effect of BTKi on collagen-induced platelet aggregation and signaling. However, it remains unclear whether the inhibition is due to an effect on BTK itself or other related kinases. Better insight into the mechanisms of platelet inhibition by BTKi may help guide the development of BTKi with a lower risk of hemorrhage.

OBJECTIVES/GOALS: The goal of this study was to examine the change in the odds of being diagnosed with metastatic cancer after the Affordable Care Act (ACA) among low-income, privately insured, nonelderly patients with newly diagnosed cancer. Low-income was defined as having income<250% FPL (federal poverty level). METHODS/STUDY POPULATION: Using Ohio cancer registry data linked with census tract-level income data, individuals aged 18-64 years diagnosed with one of the 15 leading cancers and reported being privately insured or uninsured were identified. Low-income patients were isolated using probability weighting, a process in which each observation was assigned a weight equal to the probability of a patient having an income <250% FPL based on the patient’s census tract of residence. Then, a multivariable logistic model was fitted to examine the independent association between the exposure (Post-ACA, years 2015-2016 versus Pre-ACA, years 2012-2013) and the outcome (metastatic versus non-metastatic disease at diagnosis). RESULTS/ANTICIPATED RESULTS: Between the Pre-ACA and Post-ACA periods, the percent uninsured in the low-income study population decreased from 14.1% to 4.5% (p <0.01). In the Post-ACA period, among those with insurance coverage, an estimated 11.7% of individuals had Marketplace coverage. After adjusting for potential confounders (sex, age, race-ethnicity, marital status, community-level income, rurality, and cancer type), individuals diagnosed Post-ACA had 5% lower odds of having metastatic disease relative to Pre-ACA (Adjusted Odds Ratio: 0.95, 95% Confidence Interval: 0.91 - 0.99, p = 0.04). DISCUSSION/SIGNIFICANCE OF IMPACT: The shift towards non-metastatic disease likely reflects increases to coverage brought on by the marketplaces. However, the shift is smaller than those observed in Medicaid enrollees, suggesting that policy refinements in the marketplaces can further improve outcomes in low-income cancer patients.

OBJECTIVES/SPECIFIC AIMS: Inpatient palliative care consultation (PCC) of terminally ill patients has been shown to improve patient’s understanding of their poor prognosis. In heart failure patients, PC improves transfer to hospice (or home with hospice) and decreases readmission rates. In patients with end-stage liver disease (ESLD), factors affecting having PCC has been studied, but the impact of PCC on ESLD readmissions has not been evaluated in a nationwide analysis in the US. In this study, among patients with ESLD, we evaluate the impact of inpatient PCC on 1) 30- and 90-day readmission rates, 2) hospital charges (cost) and length of stay (LOS) during subsequent 30- and 90- day readmission. METHODS/STUDY POPULATION: All ESLD hospitalizations within the first nine months of the National Readmissions Database (2010-2014) were used in this study, to allow up to 3 months to follow up. Frequencies and yearly trends of all-cause 30- and 90-days readmissions, and of PCC referral were computed. A propensity-based greedy-algorithm was used to match (1:1) patients with PCC to those without PCC (no-PCC), to create a pseudorandomized clinical study. Comparing PCC to no-PCC, generalized estimating equations were used to estimate the adjusted odds (AOR) of 30- and 90-day readmissions, and of cost and LOS during subsequent readmissions (SAS 9.4) RESULTS/ANTICIPATED RESULTS: In the United States, from January 1st to September 30th of years 2010-2014, there were 67,271 (approximating 150,396 patients) individual records of ESLD patients who survived index hospitalization. The average annual rate of PCC was 5.4%, which steadily increased from 3.84% to 6.50% over the years (p-trend <0.0001). The average 30- and 90-day readmissions rate were respectively 34.9% and 52.3%, and both remained relatively unchanged over the years (p-trends: 0.1948 & 0.5277). After matching, index PCC was associated with 68% decreased odds for 30 day readmissions (AOR: 0.32[0.28-0.37], p-value < 0.0001). When subsequently readmitted within 30 days, previous PCC resulted in a 17% shorter stay (5.7- vs. 6.9- days, p-value:0.0014) and 30% decreased cost ($47,612 vs. $68,043, p-value:<0.0001). Similarly, index PCC was associated with 74% decreased odds for 90 day readmissions (AOR: 0.26[0.24-0.29], p-value<0.0001). With subsequently readmission within 90 days, previous PCC resulted in a 17% shorter stay (5.7- vs. 6.9- days, p-value:0.0013) and 30% decreased cost ($47,520 vs. $68,016, p-value:<0.0001). DISCUSSION/SIGNIFICANCE OF IMPACT: Patients with ESLD who received PCC had a significantly lower rate of all-cause 30- and 90- day readmissions, and consumed fewer resources (hospital stay and cost) during subsequent readmissions. Although PCC resulted in a less futile use of health care resources, its adoption is still remarkably low among ESLD patients. Studies are needed to understand the barriers to PCC and to increase its use.

Objectives/Goals: Objectives/Goals (300 characters): Sarcopenia is a progressive skeletal muscle disorder associated with adverse outcomes. There is a gab of having objective measures upon performing interventions in patients with muscular dystrophies. The object of the present study is to describe the severity of sarcopenia in DMD patients in Puerto Rico. Methods/Study Population: Methods/Study Population (700 characters): Forty to 30 patients with DMD who are followed in MDA Care Center in the “Instituto De Rehabilitacion del Caribe.” Diagnosis will be confirmed with genetic testing and/or muscle biopsy. Lean muscle mass will be measured with a Whole Body Dexa (WBD) in a Nuclear Medicine Lab. Hand grip, elbow flexor, and knee extensor muscles strength will be measures with an isometric dynamometer. Patients’ functionality will done using the North Star Ambulatory Assessment scale and Brook and Vignos scales, which have been validated for patients with DMD and neuromuscular disease respectively. Correlations will be made with lean body mass (independent variable) and muscle strength and functionality (dependent variable). Results/Anticipated Results: Results/Anticipated Results (700 characters): We expect to find severe sarcopenia in patients with DMD in PR and that it will be more severe with older age. There will be a direct correlation between lean muscle mass and muscle strength, and functionality in DMD patients. Discussion/Significance of Impact: Discussion/Significance of Impact (300 characters): The findings of our study can help us to explore the possibility that Whole Body DEXA can serve as a potential biomarker for future studies since there is a need to develop noninvasive biomarkers that correlate with disease progression and interventions in DMD patients.

Objectives/Goals: We have shown that parathyroid hormone-related protein (PTHrP) is enriched at the LIFR promoter in breast cancer cells and inhibits the expression of dormancy-associated genes including LIFR. The objective of this study is to define where all PTHrP binds DNA and identify pathways that are regulated by PTHrP that promote breast cancer colonization of the bone. Methods/Study Population: In this study, we use human estrogen receptor-positive MCF7 breast cancer cells which we and others have reported lie dormant in the bone. MCF7 cells were engineered to express either PTHrP with an HA-tag (MCF7P), or a vector control (MCF7V). We use Cleavage Under Targets and Release Using Nuclease (CUT&RUN), a method of mapping protein-DNA interactions, to define where PTHrP binds DNA. Here, an HA-specific antibody identifies regions of DNA that are bound to PTHrP in MCF7P cells compared to MCFV cells. Next, we perform DNA sequencing and gene set enrichment analysis (GSEA) on genes identified by CUT&RUN to identify pathways that are regulated by PTHrP. These experiments will determine how PTHrP regulates dormancy and breast cancer colonization in the bone. Results/Anticipated Results: We completed IgG (-control), H3K4me3 (+ control), and HA (PTHrP) CUT&RUN on MCF7V and MCF7P cells, and submitted DNA for sequencing. This study will define where PTHrP binds the genome and identify pathways regulated by PTHrP. Previously, through ChIP-qPCR we showed that PTHrP binds the LIFR promoter to repress LIFR expression. Given this result, we expect that PTHrP binds to the promoters of dormancy-associated genes including LIFR in MCF7P cells compared to MCF7V cells. PTHrP may be involved in regulating other processes besides dormancy to induce expansion of breast cancer cells in the bone, so we will use GSEA to identify pathways that are altered in MCF7P cells when PTHrP is over-expressed compared to MCF7V cells. Together, this will define how PTHrP regulates gene expression of bone metastatic breast cancer cells. Discussion/Significance of Impact: This study will unveil mechanisms of metastatic breast cancer expansion in the bone by defining where PTHrP binds the genome to regulate gene expression. These findings will reveal therapeutic vulnerabilities that will be used to target bone-disseminated tumor cells to prevent lethal recurrence.

Every research study that includes volunteer participants requires safety assurances in proportion to the risks of the study. Investigator-initiated clinical research can present unique regulatory challenges particularly for studies with a risk profile that warrants more oversight than minimal risk but less than for large, commercial, or high-risk research. The use of an independent safety officer (ISO) offers a middle way of right-sizing oversight to match the risk. ISOs are clinicians or researchers with relevant expertise who are independent of the investigator and the research study. Their relationship to the study is defined by a formal charter which is aligned with the protocol and Data and Safety Monitoring Plan to address the oversight process, responsibilities of the ISO, and clearly describe the variables to be monitored. The ISO responsibilities include reviewing safety data, adverse events, recruitment, demographics, study progress, data quality, protocol changes, and any new scientific information that pertains to the trial. Finally, the ISO reports in their review on any significant findings may propose modifications to the study or a need to stop the trial.

Implementation science offers a compelling value proposition to translational science. As such, many translational science stakeholders are seeking to recruit, teach, and train an implementation science workforce. The type of workforce that will make implementation happen consists of both implementation researchers and practitioners, yet little guidance exists on how to train such a workforce. We—members of the Advancing Dissemination and Implementation Sciences in CTSAs Working Group—present the Teaching For Implementation Framework to address this gap. We describe the differences between implementation researchers and practitioners and demonstrate what and how to teach them individually and in co-learning opportunities. We briefly comment on educational infrastructures and resources that will be helpful in furthering this type of approach.