The International Study of Comparative Health Effectiveness with Medical and Invasive Approaches (ISCHEMIA) found that there was no statistical difference in cardiovascular events with an initial invasive strategy as compared with an initial conservative strategy of guideline-directed medical therapy for patients with moderate to severe ischemia on noninvasive testing. In this study, we describe the reasons that potentially eligible patients who were screened for participation in the ISCHEMIA trial did not advance to enrollment, the step prior to randomization. Of those who preliminarily met clinical inclusion criteria on screening logs submitted during the enrollment period, over half did not participate due to physician or patient refusal, a potentially modifiable barrier. This analysis highlights the importance of physician equipoise when advising patients about participation in randomized controlled trials.

Older adults have largely been excluded from health research despite bearing a disproportionate disease burden. The Community Engagement Studio (CES) model, initially developed at Vanderbilt University in 2009, allows potential research participants to help shape research to promote greater inclusion. The University of Pittsburgh adapted the CES model for older adults (OA-CES). Tailored specifically to older adults, OA-CES addresses underrepresentation in research by gathering valuable feedback that allows investigators to make research more accessible and relevant to older people. An OA-CES toolkit will help in adapting the model in other research areas to close the gap in research inclusion.

The Deep South has the highest rates of obesity, diabetes, and hypertension in the nation, with marked disparities by race, socioeconomic status, and rurality. The mission of the Forge AHEAD Center is to improve health outcomes and reduce the burden of cardiometabolic diseases across the Deep South, with a particular focus on the prevention and treatment of these chronic diseases. Forge AHEAD is unified thematically by its application of a precision public health approach across the care continuum, defined as delivering the right intervention to the right population at the right time. This approach acknowledges the importance of context and individual beliefs and preferences, as well as the need for multi-level and multi-domain interventions to achieve health improvements. The Center brings together an interdisciplinary team of investigators from 4 institutions in 3 contiguous states in the region, as well as non-academic partners to extend cardiometabolic research into real-world community and clinical settings. Given the high prevalence of cardiometabolic diseases in the Deep South and the strong research base of the partnering institutions, the Forge AHEAD Center is ideally situated to inform research, clinical care, and policy to improve health outcomes in a region of substantial need.

OBJECTIVES/GOALS: Minor procedures are anxiety-provoking and/or painful for children. Virtual reality (VR) is an emerging technology in medicine and largely has been used as an adjunct to analgesia and opioids. This study reviews the institutional use of VR in lieu of pharmacology and general anesthesia (GA) to perform minor procedures in a pediatric population. METHODS/STUDY POPULATION: A retrospective chart review was performed on all patients that presented to our institution from 2019 to 2022 for hormone implant placement, exchange, or removal with VR distraction. Demographic and procedure information was recorded. The primary outcome was successful procedure completion without requiring pharmacologic sedation or analgesia. RESULTS/ANTICIPATED RESULTS: A total of 111 patients underwent the following minor procedures with VR only. 14 patients underwent more than one procedure resulting in a total of 126 procedures. The mean age was 11.3 ± 3.6 years. 43 patients were female, 23 were female to male, 6 were non-binary, 7 were male, and 32 were male to female. 58% had private insurance. The most common diagnosis was precocious puberty (54%) followed by gender dysphoria (46%). The most common procedure was implant placement (72%). 69% of procedures were performed in the clinic and 31% in a procedural room. All procedures were completed without requiring sedation or GA. None of the patients required intravenous catheter placement for the procedure. No intra-procedural complications were recorded. DISCUSSION/SIGNIFICANCE: Despite the current trend toward minimizing GA and sedation in children, there is no widely accepted substitute. VR is a feasible option that can spare children from sedation or GA for minor procedures. This can enable procedures to be transitioned into more resource efficient settings and improve pediatric patient experience.

OBJECTIVES/GOALS: This project aimed to investigate differences between isometric and isotonic systems for low and high frequency contractions. Intracellular changes in pH caused by the rate of exercise differ in their effect on PCr. The project assessed the reproducibility between these exercise methods. METHODS/STUDY POPULATION: Four subjects were studied (2F/2M, avg. age 27 years). 31P-MRS spectra were measured in the vastus lateralis (VL) using a 3T MRI scanner using a 31P/1H surface coil. In the isometric studies, the (supine) subjects legs were supported by a wedge pad and restrained by straps over the hips and knee with a resistance strap over one ankle. In the isometric studies, the (prone) subjects legs were fit into machine designed to induce isometric exercise. There were 4 exercise bouts that differed by exercise type and frequency and, consequently, duration. 31P-MR spectra were acquired from a 25mm slab of leg positioned parasagittal along the VL. Analyses were done using jMRUI and R. RESULTS/ANTICIPATED RESULTS: With an increase in sample size, we expect differences between recovery time constants, tau, of PCr between the different exercise frequency groups alluded to by other studies investigating recovery rates in bicep skeletal muscle. From the preliminary data, we obtained large limits of agreement of tau = 0.053 s-1 & tau =-0.074 s-1 from using Bland-Altman analysis. As we increase our sample size, we anticipate that we will see differences of PCr recovery rates between the exercise types as well. DISCUSSION/SIGNIFICANCE: Current studies that investigate the differences in PCr recovery rates between different metabolic states differ in the exercise methods employed on subjects. This study aims to standardize PCr recovery assessment in the VL for more interpretable and applicable assessments of metabolic function.

OBJECTIVES/GOALS: The purpose of this work is to assess antibiotic prescribing for inpatients with community-acquired bacterial pneumonia (CABP) due to methicillin-resistant Staphylococcus aureus (MRSA) in the All of Us database. The goal of this research is to determine if different subgroups are more or less likely to receive anti-MRSA antibiotics. METHODS/STUDY POPULATION: This is a retrospective cohort study of inpatients with CABP due to MRSA from 2/1/2011 to 7/1/2022 in the All of Us database. Cases will be excluded for other treatment settings, other pathogens, and other types of pneumonia. Patients will be stratified by age, sex, race, and ethnicity. The proportion of patients who received anti-MRSA antibiotic therapy will be compared within groups with the chi-square statistic. Significant associations between patient characteristics and anti-MRSA prescribing (p < 0.05) will be assessed using multivariate logistic regression, with subgroup as the independent variable, anti-MRSA prescribing as the dependent variable, and divergent baseline characteristics as potential confounders. Odds ratios (OR) and 95% confidence intervals (95% CI) will be calculated. RESULTS/ANTICIPATED RESULTS: Previous research by our group has demonstrated differences in guideline-concordant, empiric antibiotic prescribing, for inpatients with CABP in the All of Us database; however, guideline-concordant empiric antibiotics for CABP do not routinely cover for MRSA. Anti-MRSA antibiotics are recommended if the patient has known MRSA or risk factors for MRSA. Investigations of disparity in anti-MRSA prescribing have been limited, especially since the abandonment of the healthcare-associated pneumonia (HCAP) categorization. Since the All of Us database contains information on CABP pathogens, we can study sub-types of CABP; therefore, we now hypothesize that the proportion of inpatients who received anti-MRSA antibiotics for CABP, due to MRSA, in the All of Us database, will differ by age, race, sex, and ethnicity. DISCUSSION/SIGNIFICANCE: This is one of the first studies to evaluate antibiotic prescribing for CABP due to MRSA in the All of Usdatabase. Identifying and understanding differences in care, such as possible discrepancies in anti-MRSA prescribing by age, sex, race, or ethnicity, is essential to develop targeted interventions to address disparities in health outcomes.

OBJECTIVES/GOALS: Clonal hematopoiesis of indeterminate potential (CHIP) is a common age-related condition that confers an increased risk of blood cancer, cardiovascular disease, and overall mortality. Larger proportions of blood cells with the CHIP mutation (clones) lead to worse outcomes. The goal of this study was to characterize CHIP clonal behavior over time. METHODS/STUDY POPULATION: While DNA biobanks have the ability to identify large cohorts of individuals with CHIP, they typically only contain blood from a single timepoint, limiting the ability to infer how CHIP clones change over time. In this preliminary study, we utilized multi-timepoint blood samples from 101 individuals with CHIP in Vanderbilt’s biobank (BioVU) to characterize clonal behavior over time. Using a CHIP gene-specific sequencing pipeline, we were able to characterize each individual’s CHIP mutation(s) and how the fraction of cells with the CHIP mutation expanded/reduced over time. By Spring 2023, we will also include ~300 additional individuals with CHIP in this study. RESULTS/ANTICIPATED RESULTS: CHIP mutations occurred 48% of the time in DNMT3A and 23% of the time in TET2, consistent with previous studies. 21% of individuals had more than one CHIP mutation. The mean difference in time between the two timepoints was 5.2 years (SD=2.9). Surprisingly, we observed both clonal expansion and clonal reduction across timepoints with 30% of DNMT3A, 0.6% of TET2, and 46% of JAK2 clones shrinking over time. The fastest average expansion was seen in TET2 clones (2% growth/year) and the slowest in DNMT3A clones (0.4% growth/year), but there was a significant amount of variation between individuals. In DNMT3A clones, there were no differences observed between loss of function mutations, missense mutations or DNMT3A R882 hotspot mutations. Clonal competition was observed in individuals with multiple driver mutations. DISCUSSION/SIGNIFICANCE: We used multi-timepoint blood samples to quantify the change in CHIP cell fraction over time on a per individual basis and observed novel clonal behavior and competition. Understanding the factors that influence the rate of CHIP progression can lead to personalized disease risk assessment for individuals with CHIP.

OBJECTIVES/SPECIFIC AIMS: Elucidate the unique challenges associated with hospital discharge planning for patients experiencing homelessness. Assess the impact of robust community partnerships and strong referral pathways on participating patients’ health care utilization patterns in an interdisciplinary, student-run hospital consult service for patients experiencing homelessness. Identify factors (both patient-level and intervention-level) that are associated with successful warm hand-offs to outside social agencies at discharge. METHODS/STUDY POPULATION: To assess the impact of participation in HHL on patients’ health care utilization, we conducted a medical records review using the hospital’s electronic medical record system comparing patients’ health care utilization patterns during the nine months pre- and post- HHL intervention. Utilization metrics included number of ED visits and hospital admissions, number of hospital days, 30-day hospital readmissions, total hospital costs, and follow-up appointment attendance rates, as well as percentage of warm hand-offs to community-based organizations upon discharge. Additionally, we collected data regarding patient demographics, duration of homelessness, and characteristics of homelessness (primarily sheltered versus primarily unsheltered, street homeless versus couch surfing, etc) and intervention outcome data (i.e. percentage of warm hand-offs). This study was reviewed and approved by the Tulane University Institutional Review Board and the University Medical Center Research Review Committee. RESULTS/ANTICIPATED RESULTS: For the first 41 patients who have been enrolled in HHL, participation in HHL is associated with a statistically significant decrease in hospital admissions by 49.4% (p < 0.01) and hospital days by 47.7% (p < 0.01). However, the intervention is associated with a slight, although not statistically significant, increase in emergency department visits. Additionally, we have successfully accomplished warm hand-offs at discharge for 71% percent of these patients. Over the next year, many more patients will be enrolled in HHL, which will permit a more finely grained assessment to determine which aspects of the HHL intervention are most successful in facilitating warm hand-offs and decreased health care utilization amongst patients experiencing homelessness. DISCUSSION/SIGNIFICANCE OF IMPACT: Providing care to patients experiencing homelessness involves working within complex social problems that cannot be adequately addressed in a hospital setting. This is best accomplished with an interdisciplinary team that extends the care continuum beyond hospital walls. The HHL program coordinators believe that ED visits amongst HHL patients and percentage of warm hand-offs are closely related outcomes. If we are able to facilitate a higher percentage of warm hand-offs to supportive social service agencies, we may be able to decrease patient reliance on the emergency department as a source of health care, meals, and warmth. Identifying the factors associated with successful warm hand-offs upon discharge from the hospital may assist us in building on the HHL program’s initial successes to further decrease health care utilization while offering increased interdisciplinary educational opportunities for medical students.

Objectives/Goals: The University of Illinois Chicago’s (UIC) Center for Clinical and Translational Science (CCTS) aimed to increase research dissemination findings to strengthen community relationships and opportunities for researchers to be in dialogue with communities. Methods/Study Population: From 2024 to present, the CCTS developed a collaboration with community and internal UIC partners connected to Auburn Gresham (AG), a predominantly African American community located in a far southside neighborhood of Chicago, to co-design a series of Community Forums. Community partners included UIC’s Neighborhood Center AG site, dedicated to supporting local stakeholders through strategic interventions, Office of Community Engagement and Neighborhood Health Partnerships (OCEAN-HP), fostering community partnerships through service, education, and research, and the Greater Auburn Gresham Development Corporation (GAGDC), a grassroots organization focused on community development in low-income communities. Results/Anticipated Results: Each community forum discussed research topics affecting the AG community by providing a safe space to foster bidirectional dialogue among community members, university researchers, and other field experts, while simultaneously bridging the gap between the community and research. Topics were identified through AG members’ community expertise and audience anecdotal/survey feedback. The forums were free and evolved to provide community resource packets and basic health screenings (e.g., blood pressure and glucose). This was an iterative engagement process (e.g., establishing relationships and trust, incorporating community feedback) to best mold the community forums to AG’s needs. The community audience and researchers/field experts have continued to express their interest in future community events. Discussion/Significance of Impact: The AG community forums have increased the interest in what research is and how to be involved among community members. Researchers/field experts have reiterated the importance of bidirectional dialogue with the community and the positive impact on research. These forums support research dissemination and community engagement.

Objectives/Goals: As translational researchers, we seek to offer more treatments to more people, more quickly. Essential to this mission, we must demonstrate trustworthiness to establish trusted with those we seek to benefit. Research Jam uses a human-centered design (HCD) research approach to achieve this. Methods/Study Population: HCD is an iterative process that focuses on understanding people’s contexts while designing products and services collaboratively with the people who will interact with and be affected by the research. This generative approach helps participants express tacit and latent knowledge – emotions, needs, and desires that are often challenging to communicate verbally. We work with individuals who have lived experience relevant to the research being conducted and use HCD activities to help people share their thoughts, concerns, and ideas. These activities are highly interactive, promote collaboration, and explore topics with respect and sensitivity. Results/Anticipated Results: At Indiana Clinical and Translational Sciences Institute, we use the AAMC’s principles of trustworthiness. These principles parallel the tenets of HCD research; thus, taking an HCD research approach naturally builds trust between the researcher and participants and offers tangible benefit to the research, the researcher, and those with lived experience who participate. We have ten years of work collaborating with 40 PIs and over 600 participants. Our experiences demonstrate that this approach can create an environment where participants feel comfortable as they share their thoughts, concerns, and ideas which influences the research that could affect their lives. Discussion/Significance of Impact: An HCD approach helps demonstrate a researcher’s trustworthiness with the population being served. This is essential to achieve a lasting impact by discovering the best solutions from the community’s perspective and identifying partners to help implement solutions.

Objectives/Goals: To evaluate disparities in diabetes outcomes between Hispanic/Latinx and non-Hispanic participants in a medically tailored meal intervention, assess effect modification by ethnicity/language, and explore cultural factors influencing intervention effectiveness for diverse T2DM populations. Methods/Study Population: This mixed-methods study, part of the Changing Health through Food Support for Diabetes RCT (n = 246), compares standard diabetes care to a 6-month medically tailored meal intervention for T2DM patients with food insecurity. It examines differences in intervention effectiveness among Latinx, Spanish-speaking participants and non-Latinx, English-speaking participants from San Francisco and Alameda counties. Quantitative measures include HbA1c, food security scores, and hospitalization rates. Qualitative interviews explore diabetes management, food access, and cultural factors. Baseline disparities are assessed using t-tests and chi-square analyses, while longitudinal changes are evaluated with mixed-effects models. Thematic analysis of qualitative data identifies emerging patterns. Results/Anticipated Results: We expect to find significant baseline disparities in glycemic control and food security between Latinx- and Spanish-speaking participants compared to their counterparts. The medically tailored meal intervention is anticipated to show differential effectiveness, with potentially smaller improvements among Latinx and Spanish-speaking groups. While we hypothesize that medically tailored meals will lead to improved glycemic control and reduced food insecurity across all groups, the magnitude of improvement may vary. Qualitative data are expected to reveal unique cultural and linguistic barriers contributing to these disparities, as well as insights into the acceptability and cultural appropriateness of the intervention. Discussion/Significance of Impact: This study will inform culturally tailored medically tailored meal interventions for Hispanic/Latinx- and Spanish-speaking populations, addressing disparities in diabetes outcomes and food security. Findings will shape “food is medicine” initiatives and policies to reduce chronic disease burden and health inequities in diverse communities.

ABSTRACT IMPACT: This unique approach has the capability to elucidate the pathological mechanisms underlying traumatic brain injuries and neurodegeneration, both separately and in concert, while simultaneously providing a semi-high throughput model for investigating potential pharmaceutical interventions: discoveries that would have major translational implications and a significant impact worldwide. OBJECTIVES/GOALS: We aim to improve our understanding of the mechanisms behind the development of neurodegenerative diseases by utilizing the link between traumatic brain injuries and demonstrated biomarkers with our innovative TBI on a chip model. With this tool, we hope to provide new pathological insights and explore potential pharmaceutical interventions. METHODS/STUDY POPULATION: E16 murine cortical networks were cultured onto reusable, optically transparent MEAs (fabricated in-house), and subjected to a clinically-relevant range (30-300g) of impact g-forces, utilizing our exciting new in vitro model of trauma (TBI on a chip) with real-time electrophysiological and morphological access. Impacts were systematically applied at varying intensities, repetitions, and time points, and fixed 24-hours post. Basic immunocytochemical techniques were used to investigate post-impact levels of acrolein, an established biomarker of both post-TBI oxidative stress and neurodegeneration (ND), and compared to procedurally and age-matched non-impact control networks. In addition, several other TBI/ND biomarker investigations are in progress (βA42, α-synuclein, and phosphorylated tau). RESULTS/ANTICIPATED RESULTS: Impact experiments revealed significant, force-dependent increases of acrolein (acrolein-lysine adducts) at 24hrs post impact, indicative of impact-linked neuronal degeneration. These changes were amplified by the following manipulations: increasing g-force exposure (30-250 g); the rapid (4-6 sec interval) application of multiple impacts (1, 3, 5 and 10x); and exposure to 40 mM EtOH (10 min duration immediately following impact). Further, we demonstrate the enhancement of injury recovery as a function of: increasing time intervals between repeated hits; Hydralazine exposure. In addition, conditioned media from maximally-impacted cultures can cause acrolein elevation when introduced to non-impact, control networks, indicating acrolein’s role as a diffusive-factor in post-TBI secondary injuries. DISCUSSION/SIGNIFICANCE OF FINDINGS: This novel approach provides unprecedented resolution, and is improving our understanding of the pathological mechanisms underlying both TBI and ND. Combined with our established in vivo models of trauma and computer modeling, we hope to better guide our translational laboratory endeavors and help improve clinical diagnoses and treatments.

OBJECTIVES/GOALS:

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

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

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

OBJECTIVES/GOALS: We will describe the community-driven development and impact of the new Community Research Capacity-Building grants from the Alliance for Research in Chicagoland Communities, Northwestern University. Communities expressed that to enter equitably into partnerships with academics they need support to build their own community research capacity. METHODS/STUDY POPULATION: ARCC Seed Grants, since 2008, included Partnership Development and Research Pilots, which are both jointly submitted by a community-academic partnership. The new Community Grants are submitted only by community partners and don’t require an academic partner. These grants, $3,000 over 6 months, support the development or strengthening of organizational or community-level research capacity. This may include assessing community capacity to lead and/or collaborate on research; building research capacity of community organizations (staff, leadership, residents), developing community infrastructure (e.g. research principles; staff research responsibilities; process for assessing/ tracking researcher inquiries; template memorandum of understanding) or community research priorities, etc. RESULTS/ANTICIPATED RESULTS: Eight ARCC Community Research Capacity-Building Seed Grants have been awarded so far as a part of three cycles of applications over 2022-23 (2 in 2022, 6 in 2023). During this time period, data has been collected during the application process, in final reports, and in informal group and individual discussions. Information about the profile of grantees (community representation, health focus, etc.), the initial impact of grants, and feedback from grantees about the positive and challenging aspects of the grants will be shared. Grantees have informally shared that the awards have helped to address concerns that many low-income communities of color have their voices are not adequately included in research and other decision-making. The poster will be co-presented by a community grant recipient. DISCUSSION/SIGNIFICANCE: To ensure that research partnerships are community-driven & equitable, it is necessary to invest in community research capacity-building. More evaluation is needed to understand the grants impact, as well as other approaches to community research capacity and leadership development. Poster will be co-presented by a community grant recipient.

ABSTRACT IMPACT: The ability to restore mitochondrial health in neurons derived from FXTAS patient-induced pluripotent stem cells by novel natural compounds is critically important to the management of patients experiencing this syndrome and other Fragile X associated disorders. OBJECTIVES/GOALS: The goal of this research is to assess the biological potency of MAM, NADA and MAM analogues’ neuroprotective capacity with respect to mitochondrial damage, and antioxidant properties that can restore mitochondrial health in patients with FXTAS. METHODS/STUDY POPULATION: To establish mitochondrial dysfunction, normal human cell lines and human induced pluripotent cells will be exposed to multiple concentrations of glucose/ glucose oxidase (GluOx) at several time points to induce varying intensities of oxidative stress. The degrees of oxidative stress will be measured by apoptosis and mitochondrial reactive oxygen species (ROS) production. N-arachidonoyldopamine (NADA), macamides (MAM) and its analogue compounds, effective against oxidative damage in mitochondria, will be used to rescue glucose oxidase induced oxidative damage in both cell lines. To test the ability of these drugs to restore mitochondrial health, cell viability and cellular superoxide production will be assessed by propidium iodide and the MitoSox fluorescence assay, respectively. RESULTS/ANTICIPATED RESULTS: We anticipate that GluOx at varying concentrations and time points will proportionally increase levels of apoptosis and mitochondrial ROS, reflective of mitochondrial damage, with the most severe dysfunction occurring at the maximum dose of 40µM and the longest duration of 72-hr exposure. Moreover, administration of NADA, MAM, and MAM analogues at seven concentrations, ranging from 10-8 to 10-5 M in half-log increments, will successfully treat the oxidative defects induced in the cell lines by decreasing apoptosis, and superoxide production, and increasing cell viability. DISCUSSION/SIGNIFICANCE OF FINDINGS: This research allows for the development of an in vitro neuronal model of FXTAS, lends flexibility to testing therapeutics, and expands the discovery of mitochondrial biomedical markers for the syndrome. Data generated should inform mechanistic studies of the relationship between mitochondrial damage and FXTAS-induce neurodegeneration.

OBJECTIVES/GOALS: Parent stress and coping impacts reward and motivation circuits during child development which influence self-regulation. One well known maladaptive coping response is alcohol or drinking-to-cope (DTC). This study assessed differences in stress and child behaviors among DTC parents as compared to non-DTC parents. METHODS/STUDY POPULATION: Baseline data was used from parents of a 2-5-year-old who were screened for a larger study assessing a mindfulness-based parent stress reduction intervention to improve healthy choices for themselves and their families. The sample included 172 parent-child dyads, mean parent age was 34.4 (6.1) years old, 56.3% white, mean child age was 3.6 (1.2) years old, 52.3% male. Subjective stress was assessed using the Perceived Stress Scale (PSS), parent-specific stress was assessing using the Parenting Stress Index (PSI), DTC was assessed using the COPE inventory, and child behaviors were assessed using the Devereux Early Childhood Assessment. To investigate the differences in stress and child behaviors between DTC parents and non-DTC parents independent samples t-tests were conducted. RESULTS/ANTICIPATED RESULTS: DTC was significantly correlated with PSS (r= 0.23, p<.01), PSI (r= 0.26, p<.01), child self-control (r= -0.16, p= .03), child attention problems (r= 0.22, p<.01), and total behavioral concerns (r= 0.16, p=.04). After excluding those who do not drink alcohol at all, we found significantly higher perceived stress among DTC parents (M= 27.83, SD= 9.79) compared to non-DTC parents (M= 23.79, SD= 8.40), t(80)= 2.02, p= .02. For children, we found significantly higher aggression scores for children of DTC parents (M= 47.16, SD= 31.69) compared to children of non-DTC parents (M=35.83, SD=25.72), t(84)= 1.83, p= .04. And greater attention problems among children of DTC parents (M= 73.97, SD= 26.77) compared to children of non-DTC parents (M= 56.71, SD= 34.09), t(84)=2.63, p=.01. DISCUSSION/SIGNIFICANCE: Stress and DTC may contribute to negative behaviors in children. An intervention designed to decrease stress and increase adaptive coping mechanism in parents could benefit health child socioemotional and behavioral development. Future analyses will examine third variable effects in the relationship between stress, coping, and child behaviors.

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

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

ABSTRACT IMPACT: Investigating the relationship between ICH volume at presentation and cortical atrophy over time may help to explain the phenomenon of cognitive impairment after ICH. OBJECTIVES/GOALS: Non-traumatic intracerebral hemorrhage (ICH) is the most common type of hemorrhagic stroke. ICH causes both mechanical as well as oxidative injury leading to neurologic deterioration. However, the relationship between ICH volume and brain atrophy is not fully understood. To that end, we aimed to investigate this relationship. METHODS/STUDY POPULATION: A retrospective cohort of adult ICH patients over a 10-year study period were studied. Demographic data, ICH cause, location, laterality, and treatment, as well as other data, were collected. DICOM files of CT or MRI at presentation and all subsequent follow-up imaging up to 5-years post-ICH were obtained. Using a novel semi-automated image segmentation tool developed in MATLAB by our group, ICH volumes were segmented. Using T1w MRI data, ipsi- and contralateral brain volumes were segmented using NeuroQuant, a fully automated software for MRI volumetric processing, for both initial and follow-up MRIs. Ipsilateral and global cerebral volume changes over time were calculated using initial and follow-up MRI images. Spearman rank correlation between volume changes and ICH volumes were calculated for each patient. RESULTS/ANTICIPATED RESULTS: 75 spontaneous ICH patients with adequate imaging follow-up and both early and follow-up MRI comparisons were considered. Of these, 14 patients had MRIs adequate for segmentation by NeuroQuant. There was a positive correlation (R^2 = 0.72, p=0.03) between ABC/2, the traditional ICH volume measuring approach, and our semi-automatic segmentation tool, with ABC/2 tending to overestimate ICH volume. There was an average of -6.51% of total brain volume loss with respect to initial brain volume at follow-up. There was a negative correlation between ICH volume and both global (Spearman rank correlation coefficient (R)=-0.714, p=0.004)) and local atrophy (R=-0.785, p=0.0009), meaning that as ICH volume increases, there is greater brain volume loss. DISCUSSION/SIGNIFICANCE OF FINDINGS: Greater ICH volume is associated with greater brain volume loss both ipsilaterally, reflected as encephalomalacia, and globally. These findings are important as encephalomalacia can result in focal neurologic deficit and other neurological symptoms over time, while global brain atrophy is associated with dementia and cognitive decline.