OBJECTIVES/GOALS: Our objective is to investigate the antibacterial and regenerative properties of a novel AHA dental coating for the prevention and treatment of deep caries (cavities). Further, we aim to investigate and compare these properties through in vivo murine models and assessment on human saliva samples and human pulpal cells collected from clinical samples. METHODS/STUDY POPULATION: In vitro antibacterial studies were performed by collecting and culturing human salivary bacteria with AHA substrates and quantifying survival of cariogenic Sm (S. mutans). In vivo, C57BL/6 mice were treated with AHA composite fillings, infected with Sm clinical isolates, and fed a high sucrose diet with cavity formation assessment after 6 weeks. To evaluate regeneration, mice were similarly given composite with AHA or MTA (standard of care) upon pulpal exposure with regeneration quantified by microCT and histological analysis of dentin bridge formation, ALP production, and odontoblast migration. In vitro, AHA substrates were cultured with MC3T3-E1 pre-osteoblast cells and dental pulp stem cells obtained from clinical samples over 21 days, with mineralization and ALP assessed indicating osteogenesis. RESULTS/ANTICIPATED RESULTS: In vivo studies have shown the reduction of cavity formation in mice treated with AHA as well as dentin regeneration upon pulpal exposure using microCT and histological image analysis. Coinciding with these findings, AHA substrates eradicated cariogenic Sm in human saliva samples and single species cultures in vitro. Further, preliminary results in vitro have shown increased mineralization of MC3T3-E1 cells when cultured with AHA substrates in comparison to uncoated substrates. We anticipate similar increased mineralization as well as increased ALP production of human pulpal stem cells from clinical samples when cultured with AHA substrates, suggesting osteogenesis. Further, we anticipate increased odontoblast migration and ALP production upon additional analysis of in vivo tissue samples. DISCUSSION/SIGNIFICANCE: This work will elucidate the antibacterial and regenerative properties of AHA dental coatings. These results further support the translation of AHA coatings into the clinic as a novel therapeutic for the prevention and treatment of dental decay, which may overcome the limitations associated with the current treatments.

OBJECTIVES/GOALS: To describe the strategies our newly created Patient Partners Program has planned for engaging diverse patients in the co-creation of our planned Patient Partner Academy. We will elicit feedback from other CTSAs about their strategies for promoting individual patient partnerships in research. METHODS/STUDY POPULATION: Our program to promote authentic patient-researcher partnerships to advance clinical and translational research is grounded in participatory approaches to maximize meaningful engagement. The process of creating our Patient Partners Academy is co-led by a patient partner and national leader in re-envisioning the role of patients in research, and a University of Michigan faculty member who advocates for patient partnerships in research. Listening sessions and community engagement studios will involve the developers of patient partner training programs, patient partners who have received research capacity training (as well as researchers and patient partners interested in collaborative research. Insights from these sessions will inform the development of learning models and curriculum content. RESULTS/ANTICIPATED RESULTS: Through individual listening sessions and community engagement studios we will collect data on the barriers and facilitators to patient partner engagement in research and the experiences and preferred learning models in patient partner training programs. Both the listening sessions and community engagement studios will be recorded, transcribed, and analyzed for common themes. We aim to answer three questions: What do patient partners need to be meaningfully engaged as equal partners across the research continuum? How can we best engage people who experience serious illness or chronic conditions and may be underserved by healthcare systems? What do researchers, faculty, and clinicians need to create trusting, collaborative relationships with patient partners? DISCUSSION/SIGNIFICANCE: Integrating patient partners into study teams accelerates innovation and translational science, increases the relevance of research findings, improves health outcomes and patient empowerment, and elevates the value of the patient perspective allowing researchers to gain a new point of view from an individual with lived experience.

OBJECTIVES/SPECIFIC AIMS: This study aims to describe factors impacting older adult OTC selection and use so they can be targeted with a community pharmacy intervention to improve older adult medication safety. The primary outcome is the characterization of the relationship between health literacy, risk perception, and OTC misuse. These results will directly inform the refinement of the community pharmacy intervention such that it is tailored more precisely to the older adult patient population. METHODS/STUDY POPULATION: This cross-sectional study administers face-to-face surveys to 72 older adults (age 65+) at three locations of a mass-merchandise chain pharmacy. This study is one component of a larger study to develop and implement a community pharmacy intervention to improve OTC safety for older adults. The Protection Motivation Theory (PMT) provides a framework for evaluating the interplay of threat and coping appraisals in the assessment of perceived risk severity and OTC misuse. Health literacy, known to contribute to coping appraisals, is measured with the Newest Vital Sign survey. The TRI-RISK model, originally used to measure risk perception of disease, was adapted and subject to rigorous cognitive interviewing and testing with 8 older adults. This adapted survey is used to measure older adult risk perception of adverse drug events. Walking interviews with older adult participants are conducted in a community pharmacy. In these interviews, participants verbalize their process of selection and use of an OTC to treat a pain, cough/cold, or sleep issue. Additional patient factors such as age, race, gender, education level, and co-morbidities are collected in a survey. Structural equation modeling is used for data analysis. RESULTS/ANTICIPATED RESULTS: It is hypothesized that poor health literacy leads to lower risk perception, which will result in increased instances of potential adverse drug events. Data collection will be complete by Spring 2019 and preliminary results will be presented. DISCUSSION/SIGNIFICANCE OF IMPACT: Few interventions have attempted to decrease misuse of high-risk OTC medications in older adults, and not one has addressed system barriers. Compared to efforts to improve prescription medication safety, efforts to decrease OTC medication misuse in community-dwelling older adults have been practically ignored. This is the first study to characterize and operationalize health literacy and risk perception of adverse drug events in the development of a system-level intervention to address OTC safety for older adults. The population aged 65 and older is expected to increase by 38.5 million from the year 2020 to 2050. Interventions tailored to target the older adult patient population are expected to have greater efficacy in improving older adult medication safety and in alleviating the significant corresponding strain on the US healthcare system.

Increasingly, basic, translational, and clinical research has become more collaborative, resulting in multi-institutional studies that involve common approaches to a central question. For multi-institutional projects that involve recombinant or synthetic nucleic acids, Institutional Biosafety Committee (IBC) review is generally required at each separate site. Duplicative review may result in both administrative costs and delays, without evidence of increased safety or protections, and investigator frustration. To address these inefficiencies, IBC leaders drafted a collaborative IBC Reliance Authorization Agreement. The Agreement allows one or more institutions to cede IBC review to a reviewing IBC that accepts the responsibility. The ability to cede IBC review, and the ability to rely on one decision on behalf of all collaborating institutions for a given protocol, removes delays in approval of multi-center protocols, and collaborating principal investigators are able to focus on research rather than administrative tasks. In the process, we found promotion of this collaborative model led to stronger connections among institutions and among IBC members. The requirement for IBC member representation from the local community, however, limits its broader dissemination; we make several recommendations to mitigate this challenge.

Long-term sequelae of severe acute respiratory coronavirus-2 (SARS-CoV-2) infection may include increased incidence of diabetes. Here we describe the temporal relationship between new type 2 diabetes and SARS-CoV-2 infection in a nationwide database. We found that while the proportion of newly diagnosed type 2 diabetes increased during the acute period of SARS-CoV-2 infection, the mean proportion of new diabetes cases in the 6 months post-infection was about 83% lower than the 6 months preinfection. These results underscore the need for further investigation to understand the timing of new diabetes after COVID-19, etiology, screening, and treatment strategies.

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.

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: 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/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 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.

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: 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: 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.