OBJECTIVES/GOALS: In this study, we implemented deformable medial modeling as a morphometric approach in first trimester placentas to characterize morphometric differences between fully automated and manual segmentations. METHODS/STUDY POPULATION: Twenty placentas from singleton pregnancies between 11-14 weeks’ gestation were manually and automatically segmented from 3D ultrasound volumes. Automated segmentations were produced by a trained convolutional neural network pipeline. Dice overlap scores and volumes were computed between manual and automated segmentations. Deformable medial modeling was applied to both manual and automated segmentations to produce the following metrics: maternal and chorionic surface areas (SA), thickness, circumference, and diameter along the generated medial surface. Placental non-planarity was also determined as the greatest medial surface height difference. A paired t-test and simple linear regression was performed between manual and automated segmentations for each shape metric. RESULTS/ANTICIPATED RESULTS: Mean placental volume measurements between manual and automated segmentations were similar, with a percent difference of 3.28% and a mean Dice overlap score of 0.85 ± 0.07. There were strong, statistically significant (p <0.01) linear correlations with chorionic and maternal SA, SA difference, thickness, circumference, medial surface diameter, and medial surface height difference. No significant differences were noted with chorionic SA, thickness, circumference, maximum medial surface diameter, or medial surface height difference. However, statistically significant differences (p <0.01-0.03) were noted in maternal SA, SA difference, and mean medial surface diameter. Despite these differences, mean percent difference for all morphometric parameters was less than or equal to 10%. DISCUSSION/SIGNIFICANCE: A deformable medial model evaluate unique global and regional shape placental features with highly correlated values between manual versus automated placental segmentations. However, clinical studies are needed to determine if minor differences would impact the clinical utility of these features as potential indicators of placental function.

OBJECTIVES/SPECIFIC AIMS: To longitudinally track emerging research collaborations and assess their development and productivity. METHODS/STUDY POPULATION: In four administrations (2011, 2013, 2015, 2017), all full- and part-time University of Rochester Medical Center faculty received an email invitation to complete a research collaborators survey. Respondents indicated whether they were involved in research, and if involved in research, identified collaborators from a drop-down list of investigators in the institution. Space was provided for write-ins. Full- and part-time status, faculty rank, and departmental affiliation was associated with each investigator. Grant data were obtained from a grant management database maintained by the institution’s Office of Research and Project Administration. Grant data included all submissions (funded and not funded), award number, award effective data, award final expiration date, funding amounts, principal investigator and co-investigators. Using Mathematica SNA software, for each year we identified collaborator dyads (including their characteristics such as inter/intradepartmental; investigator characteristics) and networks (e.g. size, density). RESULTS/ANTICIPATED RESULTS: On average, 1800 (range 1730-2034) full- and part-time faculty received email invitations to complete the survey. An average of 403 respondents (range 385-441) completed the survey each administration. While the response rate seems low, the survey was distributed to every faculty member regardless of their primary appointment. Thus it included a large number of individuals whose role is exclusively clinical. Grant data included 4429 awards received between 2011 and 2018, involving 1395 investigators as principal or co-investigators. Survey respondents naming collaborators ranged from 233 to 280 (average 257) with 1594 to 2265 (average 1988) collaborations named each year. Overall density increased from.0204 in 2011 to.0342 in 2017. Density within the group of female investigators increased from.0219 in 2011 to.0412 in 2017. Within the group of male investigators, density increase from.0226 to.0333 in the same time span. Analysis by rank, changes over time and those with grant funding is underway. DISCUSSION/SIGNIFICANCE OF IMPACT: This methodology captured a consistent number of collaborations over an 8 year period. Analyses reveal network growth over time and of increasing heterogeneity (by gender). Analyzing research networks overtime provides an important metric to assess how research networks evolve and devolve and the characteristics of those that grow or stagnate. Further these analyses can demonstrate the impact of support provided to networks or teams by the CTSI, department or other institutional mechanism.

Objectives/Goals: Explore and compare the functional mechanisms of song-based exercises compared to speech-language pathology exercises for dysphagia. The long-term goal is to increase patient outcomes through song-based programs that are accessible, enjoyable, and personalizable. Methods/Study Population: We will pilot the use of combined electroencephalography (EEG) and electromyography (EMG) technologies to analyze both central and peripheral contributors to laryngeal control in a cohort of healthy individuals. This approach provides detailed insight into the coordination between neural and muscular activity, which will serve as a baseline for future studies in clinical populations. Song-based vocal exercises will be compared with standard dysphagia exercises prescribed by speech-language pathologists to assess their mechanistic differences. Results/Anticipated Results: We anticipate identifying specific song-based tasks, such as variations in pitch, rhythm, and intensity, which differentially impact laryngeal musculature. Additionally, we will localize neural activation hotspots using EEG during these tasks, providing a more comprehensive understanding of how song-based therapy influences both peripheral and central mechanisms. Discussion/Significance of Impact: This project will lay the groundwork for developing evidence-based song-based therapies for dysphagia, providing an alternative to traditional SLP exercises. By creating an engaging therapeutic program, we aim to reduce dysphagia’s healthcare burden, including aspiration events, healthcare costs, and related mortality.

ABSTRACT IMPACT: Learning Health System (LHS) Science that trains postdoctoral scholars from diverse professional backgrounds in methodological and professional skills to implement rigorous research in health care systems and populations, and to disseminate the findings of such research to improve healthcare delivery OBJECTIVES/GOALS: The WFUHS CTSA developed an innovative TL1 in Learning Health System (LHS) Science that trains postdoctoral scholars from diverse professional backgrounds in methodological and professional skills to implement rigorous research in health care systems and populations, and to disseminate the findings of such research to improve healthcare delivery METHODS/STUDY POPULATION: Training is centered around formal LHS science coursework and mentored research projects that address a pressing health system issue. Projects are closely guided by a primary mentor and a multidisciplinary mentoring team. Program mission and competencies were carefully evaluated in a competency-course matrix to design new courses for the LHS Certificate and MS program in Translational and Health System Science (THSS). Course domains include biomedical informatics; improvement and implementation science; system science and organizational change management; stakeholder engagement, leadership, and research management; ethics of health systems research; and health systems research methods. Scholars set up Individual Development Plans (IDP) and self-assess 7 domains of LHS core competencies. RESULTS/ANTICIPATED RESULTS: The first professionally diverse group of scholars (MD, PhD, DrPH, PharmD) began the program in Summer 2020; onboarding was conducted virtually. Scholars currently conduct most of their research and training in a virtual, synchronous format. Each developed a detailed IDP and LHS research project, which was reviewed by their LHS mentoring teams (includes a primary mentor, co-mentor, TL1 core faculty mentor, peer mentor, and health system mentor). Coursework, leading to a 1-year certificate or 2-year MS degree, was selected based on individual background and career goals and was begun in August 2020. In addition to the courses noted above, Scholars are embedded in a healthcare improvement team. We use the process of a LHS and hold weekly TL1 leadership meetings to swiftly address challenges and implement improvements DISCUSSION/SIGNIFICANCE OF FINDINGS: We envision that TL1 Scholars will build independent LHS research programs or lead health system innovation. Program evaluation includes assessments of Scholar fluency in LHS competencies and attainment of key milestones during and after training. Annual TL1 faculty retreats will address program fidelity and implementation of program refinements

OBJECTIVES/GOALS: NSF often requires cross-disciplinary team composition to be competitive for funding. To what extent do research teams have multidisciplinary authorships after they win an award, given that awards are not contracts? We examined the quantity and quality of multidisciplinary collaboration of NSF-funded teams before and after receiving their award. METHODS/STUDY POPULATION: Our sample was 150 PIs and Co-PIs (67% male) from 58 NSF-funded EAGER (EArly-concept Grants for Exploratory Research) grants between 2013 and 2019. Using publicly available information, we collected the number of conference papers, publications, and grants PIs/co-PIs produced with each other (all PIs and co-PIs in a team or a partial subset). Based on Ph.D. fields, we also cataloged whether the combination of PIs/co-PI authors on outputs represented unidisciplinary or multidisciplinary collaboration after their NSF award. Multidisciplinary collaboration consisted of multidivisional (Ph.D. disciplines across NSF divisions, e.g., political science and cognitive psychology) or multidirectorate (Ph.D. disciplines across NSF directorates, e.g., psychology and engineering) authorship. RESULTS/ANTICIPATED RESULTS: Of the 74% of PI and co-PI teams who collaborated after their EAGER award, almost 9 out of 10 chose to work with a collaborator from a different discipline, and almost 8 out of 10 chose to work with a researcher from an extremely diverse discipline from their own (e.g., computer science and psychology). Research on interdisciplinary teams largely emphasizes the challenges and problems they face but the current research demonstrated that 90% of the sample chose to continue working together across disciplines after EAGER awards. Therefore, future research should dedicate more attention to the nontangible benefits members receive in interdisciplinary teams. Moreover, quality measures revealed higher H-indices for multidisciplinary than unidisciplinary journals and conferences. DISCUSSION/SIGNIFICANCE: Our archival results revealed that NSF EAGER grants are having their intended effect of being a catalyst for 1) continued multidisciplinary (and especially multidirectorate) collaboration) and 2) high-quality multidisciplinary publication and conference output. These results have contributed to NSF policy changes to reinstate the EAGER grant.

Clinical and Translational Science Award (CTSA) hub websites are a critical communication gateway to assist the clinical and translational science community and promote CTSA hub offerings. The objective of this funded pilot project was to create a website and online database for the CTSA consortium that allows users to conduct structured searches among the 50 + CTSA hub websites. The result is CTSA Search Solutions, an online, searchable database that includes access to 50 + CTSA hub websites with 80+ structured search term options and over 800 links collected, organized, and published. Hubs can be searched by name and filtered by a specific CTSA topic, state, region, or even number of years funded to make detailed comparisons with the data identified. The home page for each hub can be accessed directly from the search page. The CTSA Search Solutions online database will allow for a wide breadth of CTSA personnel (core leads, researchers, administrators, communicators, and evaluators) to find consolidated information to learn about specific CTSA hub program highlights, as well as conduct research into program hub outputs and best practices across the nationwide CTSA consortium.

OBJECTIVES/GOALS: Highly Active Antiretroviral Therapy (HAART) is beneficial for managing HIV infection, however the long-term use of HAART may be problematic for healthy weight maintenance. The aim of the study was to investigate the association of race, weight status, and co-morbidities among individuals with HIV. METHODS/STUDY POPULATION: Self-reported data from 283 participants who completed the Symptom Checklist, the Co-Morbidity Questionnaire, and the Sociodemographic Questionnaire were included in the data analyses. Data were analyzed using Latent Class Analysis on JMP 13. Approximately 50% of participants self-identified as Black, 69% as male, and 35% as having AIDS. Participants’ age ranged from 25 to 66 years (mean age = 43.70 years, SD = 8.14). Participants were grouped by race (self- reported Black or non-Black), and then each group was clustered based on the top three most prevalent symptoms. The clusters identified were least symptomatic, weight gain, and weight loss. RESULTS/ANTICIPATED RESULTS: The non-Black weight gain cluster reported a higher incidence of AIDS (70.6% vs 38.2%), nausea (70.6% vs 17.6%), diarrhea (70.6% vs 26.5%), and shortness of breath (58.8% vs 20.6%) compared to the Black weight gain cluster. The Black weight loss cluster reported a higher incidence of cardiovascular symptoms including chest palpitations (42.2% vs 2.7%), chest pain (44.4% vs 8.1%), and shortness of breath (73.3% vs 35.1%) and a higher incidence of all GI symptoms with the most prominent being diarrhea (71.1% vs 48.6%) compared to the non-Black weight loss cluster. DISCUSSION/SIGNIFICANCE OF IMPACT: Future studies supporting these results will assist practitioners to target treatments that may prevent adverse health outcomes for individuals with HIV on HAART. Further studies will also assist with setting standards that allow practitioners to provide personalized care for individuals with HIV on HAART.

OBJECTIVES/GOALS: Using multi-state discharge data, to identify predictors of frequent emergency department (ED) use among the homeless patients seen in emergent care, and to compare frequent versus less frequent homeless ED users for their risk of serious health services utilization outcomes. METHODS/STUDY POPULATION: Based on the State Emergency Department Database and the State Inpatient Database, homeless individuals (n = 88,541) who made at least one ED visit in four states (Florida, Maryland, Massachusetts, and New York) in 2014. In this retrospective cross-sectional analysis, patient-level demographic and clinical factors were assessed as predictors for increased ED use. Risks of opioid overdose, opioid-related hospital admission/ED visit, in-hospital mortality, mechanical ventilation, and number of hospitalizations were compared between individuals with 4 or more vs. 2-3 vs. 1 ED visit(s), adjusting for potential confounders including hospital fixed effects (allowing for within hospital comparisons). RESULTS/ANTICIPATED RESULTS: Higher rates of ED use were associated with Medicare coverage <65; primary diagnosis of alcohol abuse, asthma, or abdominal pain; and co-morbidity of alcohol abuse, psychoses, or chronic pulmonary disease. Individuals with ≥4 visits had significantly higher adjusted risk of opioid overdose (3.7% vs. 1.2% vs. 1.0%), opioid-related hospitalizations/ED visits (17.9% vs. 8.5% vs. 6.6%), mechanical ventilation (9.8% vs. 7.0% vs. 4.7%), and greater # of hospitalizations (3.2 vs. 1.3 vs. 0.8) compared to individuals with 2-3 or 1 ED visit. Individuals with ≥4 and 2-3 ED visits had similar but increased risks of in-hospital mortality compared to individuals with 1 ED visit (2.8% vs. 2.8% vs. 2.3%). DISCUSSION/SIGNIFICANCE OF IMPACT: Homeless patients who were high ED users were more likely to be hospitalized and have other adverse outcomes. These findings encourage targeted interventions (i.e. housing) for the high-utilizer homeless population to reduce the burden of serious outcomes and costs for the patient and society.

OBJECTIVES/GOALS: FXT/AS is a devastating, rare neurological syndrome that negatively impacts movement and cognition and is suspected to induce mitochondria dysfunction. Currently, no effective pharmacological treatments for FXTAS exist. The goal is to restore mitochondrial viability using endocannabinoid-like compounds in a cell culture model of FXTAS. METHODS/STUDY POPULATION: To establish a cell model of mitochondrial dysfunction, fibroblast baby hamster kidney (BHK-21) cell lines were treated with glucose oxidase (GluOx) at varying concentrations and times. Mitochondrial viability was assessed by the colorimetric Janus B Green Assay, which stains the mitochondria and enables assessment of cell numbers and the presence of oxygen in anchorage-dependent cell culture. Upon establishing this model of mitochondrial dysfunction, we next investigated the ability of three novel mitochondrial antioxidants (e.g., macamides) to protect mitochondrial viability. RESULTS/ANTICIPATED RESULTS: GluOx treatment of BHK-21 cells caused a dose- and time-dependent increase in oxidative stress. The data demonstrated significant disruption in the morphology of BHK-21 cells at a high glucose concentration, i.e., 40 nM, between 2 and 24 hours post-exposure. The morphology data were confirmed by the Janus B Green colorimetric assay. In examining the effects of glucose on mitochondrial viability, we demonstrated that at 15, 30, 35, and 40 nM, glucose significantly decreased mitochondria viability compared to the untreated, with 40 nM having the greatest effect. Under these conditions of mitochondrial dysfunction, co-incubation of the cells with the 0.5 uM MAM69 macamide attenuated the GluOx-induced increase in oxidative stress, with 0.5 uM MAM69 alone showing no effect on mitochondria viability. DISCUSSION/SIGNIFICANCE: This study illustrates the efficacy of macamides, natural occurring endocannabinoid like-compound, as novel prognostic and therapeutic candidates in the treatment of mitochondrial dysfunction that is associated with FXTAS. The use of BHK-21 fibroblast cells provides a rapid screening model to test for pharmacological therapeutic efficacy.

OBJECTIVES/GOALS: The trabecular meshwork (TM) and Schlemm’s canal (SC), located within the iridocorneal angle (ICA), form the main outflow pathway and a major target for glaucoma treatments. We characterized the human ICA in vivo with Optical Coherence Tomography (OCT) imaging using a customized goniolens and a commercial OCT device (Heidelberg Spectralis). METHODS/STUDY POPULATION: Imaging these structures is difficult due to the optical limitations of imaging through the cornea at high angles. Therefore, a clinical gonioscopy lens was modified with a 12mm plano-convex lens placed on its anterior surface to focus light on the ICA structures, and capture returning light. Each subjects’ eye was anesthetized with 1 drop of Proparacaine 0.5%. The goniolens was coupled to the eye with gonio-gel and it was held by a 3D adjustable mount. OCT volume scans were acquired on 10 healthy subjects. The linear polarization of the OCT was rotated with a half-waveplate to measure dependence of the ICA landmarks on polarization orientation. RESULTS/ANTICIPATED RESULTS: The TM was seen in 9 of 10 subjects. Polarization rotation modified the brightness of the band of extracanalicular limbal lamina (BELL) and corneoscleral bands due to the birefringent nature of the collagenous structures, increasing the contrast of SC. SC width was 99 ± 20µm varying in size over space, including a subject with SC narrowing in the inferior-temporal quadrant. DISCUSSION/SIGNIFICANCE: This clinically suitable gonioscopic OCT approach has successfully been used to image the human ICA in 3D in vivo, providing detailed characterization of the TM and SC as well as enhancing their contrast against their birefringent backgrounds by rotating the polarization of the imaging beam.

Objectives/Goals: The creatine (Cr) system is impaired in Alzheimer’s disease (AD). Data show that creatine monohydrate (CrM) supplementation may improve AD symptoms in AD mouse models, but no human studies have been reported. Thus, we investigated whether an eight-week CrM supplementation was feasible and associated with increased brain creatine in patients with AD. Methods/Study Population: Twenty participants with probable AD were allocated to an open-label, eight-week intervention of 20 g/day CrM. Fasting blood draws were taken at baseline, 4-, and 8-week visits to measure serum creatine (Quest Diagnostics). 1H magnetic resonance spectroscopy was performed at baseline and 8-week visits to measure brain Cr as a ratio to unsuppressed water. Self-reported compliance (with assistance from study partners) was assessed with daily CrM trackers. The mean compliance percentage across all participants was used to describe overall compliance with the intervention. We used paired t-tests to analyze the mean changes in serum Cr levels from baseline to 4- and 8-week visits and the mean change in brain Cr from baseline to 8-week visits. Statistical significance was set at p<0.05. Results/Anticipated Results: Participants were 65% male with a mean age of 73.1±6.3 years. All participants completed the study, with 19 out of 20 achieving the dose compliance target of ≥80%. The mean self-reported dose intake was 90%. Serum Cr levels were significantly increased at 4- and 8-week visits compared to baseline (0.6±0.4 mg/dL vs. 14.0±9.9 mg/dL and 15.0±13.6 mg/dL, respectively; p<0.001). Brain Cr levels also significantly increased (330.5±36.80 i.u. vs. 366.9±57.52 i.u., p<0.001). Discussion/Significance of Impact: We are the first to demonstrate that 20 g/day of CrM for eight weeks is feasible and associated with increased brain Cr in patients with AD. Our findings support further investigation of brain target engagement of CrM and its efficacy in AD. With AD cases expected to rise, CrM could serve as an effective, affordable therapeutic to slow AD progression.

OBJECTIVES/SPECIFIC AIMS: Cardiac radiation exposure following anti-cancer (CA) thoracic radiotherapy (RT) treatment increases risk of heart failure in a dose-dependent manner with a predominantly restrictive cardiomyopathy phenotype and is characterized by a diffuse fibrosis within the myocardium. The peak oxygen pulse (O2Pulse) determined at cardiopulmonary exercise testing (CPET) is the quotient of oxygen consumption (VO2) divided by the heart rate (HR) at peak exercise. Through deduction of the Fick equation (VO2 = cardiac output (CO) x arteriovenous oxygen difference) it provides a noninvasive estimate of the stroke volume response to exercise. Knowledge of the relationship between cardiac radiation dose and O2Pulse may provide mechanistic insight into the cardiac reserve of the CA survivor following thoracic RT. METHODS/STUDY POPULATION: Patients without a history of cardiovascular disease with a history of thoracic RT for CA treatment with significant incidental heart exposure (≥5 Gray (Gy) to ≥10% of the heart volume) underwent treadmill CPET to determine cardiorespiratory fitness and cardiac magnetic resonance (CMR) imaging to quantify central hemodynamics and for myocardial tissue characterization. The mean cardiac radiation dose (MCRD) and %volume of heart dose was determined from dose-volume histograms reflective of the dose contributions from all RT treatments for each patient. The oxygen pulse (milliliters (mL) of O2 per heart beat) was determined by dividing the absolute VO2 by the HR (beats per minute, bpm) at peak exercise and reported as %-predicted values to account for age and gender differences. Data are reported as number (%) or median (interquartile range). A stepwise multivariate linear regression model was created from significant univariate RT and CMR variables to determine independent predictors of %O2Pulse. RESULTS/ANTICIPATED RESULTS: Thirty patients (age = 63 [57-67] years, 18 [60%] female, 2.0 [0.1-28.7] years since completion of RT) underwent study procedures. The peak VO2=1376 mL·min-1 (62% of predicted) and peak HR = 150 (122-164) bpm resulted in a peak O2Pulse of 9.2 mL/beat (82% of predicted). The MCRD = 5.6 [3.7-17.8] Gy was inversely associated with %O2Pulse at univariate analysis (R = −0.514, p < .01), but was not retained at multivariate analysis. The CMR-derived CO ([4.9 (4.09-5.90) Liters/minute], β = +.374, p < .01), CMR-extracellular volume ([ECV, 26.9 (24.8-29.2)%], β = −.536, p < .01), and volume of the heart exposed to ≥30 Gy ([2.5 (0-15.0)Gy], (β = −.345, p = .01) were retained in the model (R2 = .709, F(3,19) = 15.438, p < .001) and were independent predictors of the %O2Pulse. DISCUSSION/SIGNIFICANCE OF IMPACT: In patients with significant heart exposure following RT, %O2Pulse (a surrogate of stroke volume response to exercise) is inversely associated with cardiac radiation dose and is related to central hemodynamics (CO) and markers of diffuse fibrosis (ECV).

OBJECTIVES/GOALS: To determine if incorporating specific laboratory values and plasma biomarkers (club cell secretory protein (CC16), matrix metalloproteinase 3 (MMP3), interleukin 8 (IL-8), protein C) to the Lung Injury Prediction (LIP) Score improves the predictive value for development of acute respiratory distress syndrome (ARDS) in ICU patients. METHODS/STUDY POPULATION: Adult patients admitted to the ICU on supplemental oxygen over baseline requirement with a LIP Score ≥6 will be included. Patients admitted to the ICU >24 hours, end-stage renal disease, decompensated heart failure, or <100 µL plasma available will be excluded. Whole blood will be collected from the core lab, centrifuged, and plasma will be stored at -80°C. Protein biomarkers will be measured using enzyme-linked immunosorbent assay. Baseline characteristics, laboratory values, ventilator parameters, and clinical outcomes will be collected from the medical record. ARDS will be defined by the Berlin criteria. Machine learning methods will be used to identify the model with the highest predictive accuracy. Area under the receiver operating characteristic curve of each model will be compared to the LIP Score. RESULTS/ANTICIPATED RESULTS: Research is in progress. Plasma samples and clinical data have been collected for 148 of the 160 samples required to achieve power. Biomarker analysis will take place after sample collection is complete. We anticipate a machine learning model incorporating laboratory values and one or more plasma biomarkers into the LIP Score will outperform the baseline LIP Score for prediction of ARDS development. DISCUSSION/SIGNIFICANCE: Delayed diagnosis and intervention contribute to poor ARDS outcomes. Current predictive models for ARDS have low accuracy and enriching these models with plasma biomarkers may increase their predictive value. Development of accurate models may facilitate earlier ARDS diagnosis and intervention as well as enrichment strategies for ARDS trials.

Objectives/Goals: The study focuses on developing a wound patch that employs a biocompatible matrix which incorporates mesenchymal stem cells (MSCs) with wound healing and antimicrobial properties, along with antimicrobial metallic nanoparticles covered with keratinocytes derived from induced pluripotent stem cells to replicate the skin’s barrier function. Methods/Study Population: In vitro experiments will be conducted to combine bacteria with MSCs and metallic nanoparticles to assess whether bacterial killing is improved by this combination. The MSCs will then be evaluated in the presence of the nanoparticles to confirm that their functionality and phenotype are not altered. To verify the cells’ functional integrity, they will undergo trilineage differentiation, surface marker phenotypic testing, and evaluation of their capacity to inhibit lymphocyte proliferation in the presence of the nanoparticles. Subsequently, this living bandage will be created using a biomatrix embedded with induced pluripotent stem cell-derived keratinocytes and tested on a canine wound model to study the impact on healing. The model will assess the rate of healing and cellular response at weekly intervals until healed. Results/Anticipated Results: The combination of mesenchymal stem cells and antimicrobial nanoparticles works synergistically to enhance bacterial killing in vitro with S. aureus. The presence of the nanoparticles in combination with MSC did not affect the ability of the MSC to undergo trilineage differentiation. We anticipate that the surface phenotype will be similarly unaffected. In addition, we expect that the presence of the nanoparticles should not interfere with the ability of MSC to suppress lymphocyte proliferation. Utilization of the wound patch in the in vivo canine wound model is expected to enhance healing and prevent infection. We expect that we will observe a shift in the cellular composition of the wound with less inflammatory cells and more M2 or wound healing anti-inflammatory monocytes. Discussion/Significance of Impact: The incidence of resistant infections with no pharmacologic therapy available are on the rise. The development of an antimicrobial living bandage that increases the body’s ability to fight off infection, while providing a barrier to reinfection would provide a new way to treat infections regardless of their acquired antibacterial resistance.

OBJECTIVES/GOALS: Antibodies play an important role in the pathogenesis of a wide range of diseases, including cancer, autoimmune diseases, and infections. There are currently no reliable methods to isolate and study specific plasma cell subpopulations as antibody production sources. We aim to develop methods to study plasma cells in high resolution. METHODS/STUDY POPULATION: We will use molecular cloning to engineer fusion proteins that would bind plasma cell proteins to study these cells based on their surface features. The first phase of our study consists of assessing the efficacy of this plasma cell isolation method in established cell lines (e.g., RPMI 8226) and also antibody-secreting cell lines that we are establishing as a part of this study. In the second phase of the study, we will assess the efficacy of this method by studying antigen-specific plasma cell populations in the bone marrow aspiration samples of 20 healthy volunteers using various assays, including ELISPOT, flow cytometry, and fluorescent microscopy. RESULTS/ANTICIPATED RESULTS: We have designed the constructs and have completed the cloning. The final plasmids have been verified using various restriction enzymes and Sanger sequencing. Following the transfection of Freestyle HEK 293F cells and isolation of respective proteins, we expect to be able to utilize these engineered proteins to differentiate various antibody-secreting plasma cells. We will use cell lines for proof-of-concept experiments and will subsequently move this method to human bone marrow samples. We expect to be able to visualize multiple specific antibody-secreting plasma cell populations using fluorescent microscopy and utilize this method to isolate them by cell sorting via flow cytometry. DISCUSSION/SIGNIFICANCE: We expect to be able to use this method to target specific plasma cell clones in the advancement of precision medicine regarding the treatment of plasma cell disorders (e.g., multiple myeloma) and also expand its use in other areas, such as antibody discovery and the assessment of the humoral immune responses in infectious diseases.

ABSTRACT IMPACT: This study will generate preliminary data to address a critical, care transition-related patient safety gap involving medication use among children with medical complexity. OBJECTIVES/GOALS: The objectives of this study are: (1) to understand care transition-related medication safety risks for children with medical complexity (CMC), and (2) through a participatory, human centered design (PD) approach, to develop an early prototype intervention to address identified safety risks. METHODS/STUDY POPULATION: The study population includes children with medical complexity (CMC), a medically fragile pediatric population with intensive healthcare needs. CMC rely on multiple and complex medication regimens and/or medical devices for optimal functioning. Parents of CMC report multiple unmet healthcare needs. For Aim 1, we will conduct observations and interviews with ˜15 clinicians as well as semi-structured interviews with ˜30 family caregivers during three care transition experiences: from Cardiac ICU to home, Neonatal ICU to home, and those between primary care/specialty clinic to home. For Aim 2, we will conduct participatory design sessions with up to 5 participants (separately for clinicians and family caregivers) from each of the three care transition settings to co-design a prototype intervention. RESULTS/ANTICIPATED RESULTS: The study is currently recruiting family caregivers of CMC for aim 1 research activities, with interviews planned to be completed in February/March 2021. Transcribed interviews will be used to inform development of patient journey maps. A patient journey map helps to visually depict healthcare services through the patient and family lens, and highlights important ‘touch points’ along the patient journey (e.g., decisions, encounters, constraints, emotional states, etc.) that shape the patient and family experience. The journey map will distill findings from qualitative data and generate a concise visual story focused on the medication use experience of CMC as they transition between the hospital and their home. Individual journey maps will also be combined to generate a consolidated journey map. DISCUSSION/SIGNIFICANCE OF FINDINGS: An-in-depth understanding of medication safety risks unique to the context of CMC care would be essential to develop interventions that are useful, scalable, and sustainable. This is even more important because current interventions are primarily adopted from adult care settings with mixed outcomes.