OBJECTIVES/GOALS: Microtubule poisons, like Taxol, are used to treat triple negative breast cancer (TNBC) and may induce lethal aneuploidy in cancer cells. Patients initially respond, but often develop drug resistance. New targeted drugs that cause aneuploidy may be a valuable approach to therapy. One potential target is the Kinesin 13 MCAK, which limits aneuploidy. METHODS/STUDY POPULATION: TCGA and GSE47561 databases were probed for MCAK expression, and data was stratified by subtype and survival statistics. Knockdown studies were performed to test whether MCAK knockdown sensitizes cells to taxanes for cell proliferation and for induction of aneuploidy. FRET and image-based screens were used to identify MCAK inhibitors from small molecule inhibitor libraries. Inhibitors were then tested for functional effects in multiple cell-based assays and for clonal growth in colony formation assays. RESULTS/ANTICIPATED RESULTS: MCAK expression is upregulated in TNBC and associated with reduced overall survival. Knockdown of MCAK caused a two-to-five-fold reduction of the IC50 for Taxol in cancer cell lines, with no change in normal cell lines. Taxol treatment or MCAK knockdown increased aneuploidy induction, with no additive effect between the two. Our small molecule screen identified three putative MCAK inhibitors, which induced aneuploidy in both taxane-sensitive and taxane-resistant cells. These inhibitors also reduced clonogenic growth, and the most potent inhibitor, C4, caused an approximate five-fold reduction in the IC50 for Taxol in cell proliferation assays. DISCUSSION/SIGNIFICANCE: MCAK can serve as a biomarker of breast cancer prognosis. MCAK knockdown or inhibition sensitizes cancer cells to Taxol without affecting normal cells, making it a potential target in combination therapy. MCAK inhibitors also reduce growth as single agents in taxane resistant lines, giving them potential use as therapies in resistant disease.

OBJECTIVES/SPECIFIC AIMS: The advent of Highly Active Antiretroviral Treatments (HAART) has allowed HIV-positive individuals to live longer in recent years. This has resulted in a higher incidence of mortalities occurring in these individuals due to cardiovascular pathologies, as opposed to deaths due to HIV. Even with long-term HAART, persons living with HIV (PLWHIV) still exhibit inflammation, which is associated with deleterious cardiovascular outcomes. PLWHIV on HAART have a higher prevalence of hypertension, which is associated with an increased risk of cardiovascular events. Moreover, chronic inflammation has been shown to be related to the translocation of microbes and endotoxins across the gastrointestinal tract. Such microbial translocation (MT) is increased in individuals with digestive disorders and their associated symptoms (e.g., diarrhea, abdominal pain, and nausea). This study aims to explore the pathologies common to both MT-induced inflammation and cardiovascular symptoms by examining the associations between gastrointestinal symptoms and hypertension in PLWHIV on HAART. METHODS/STUDY POPULATION: The sample included 351 PLWHIV on HAART. Pre-existing de-identified data were analyzed. Sample demographics included 56.98 % African Americans, 41.31% Caucasians, ages 20–66 years (mean age=43.65years), 21% female, 89% male, HIV viral load, CD4 counts. Self-reported data from the Symptom Co-Morbidity Questionnaire and Socio-demographic questionnaire were analyzed with SPSS v.24. RESULTS/ANTICIPATED RESULTS: In total, 86 PLWHIV (24.50%) stated that they have hypertension; 39 subjects (45.3%) reported having diarrhea, 30 subjects (34.8%) reported nausea, and 12 (13.9%) reported constipation and vomiting. Among ethnicities with hypertension and gastrointestinal symptoms, African Americans compared with Caucasians had a higher percentage of diarrhea (28% vs. 17%), nausea (21% vs. 11%), constipation (11% vs. 2%), and vomiting (8% vs. 5%). Women compared with men reported a higher percentage of nausea (28% vs. 24%) and constipation (8% vs. 6%). Men compared with women reported a higher percentage of diarrhea (38% vs. 7%) and vomiting (8% vs. 5%). DISCUSSION/SIGNIFICANCE OF IMPACT: These data support the need for targeted screening to include both blood pressure and associated gastrointestinal symptoms. Further studies supporting these results may assist practitioners to target treatments that may prevent cardiovascular comorbidities.

OBJECTIVES/SPECIFIC AIMS: 1. Identify the extent of CD implementation for trained childcare teachers. 2. Explore teacher perspectives on the impact of CD. 3. Explore teacher perspectives on barriers and facilitators to full implementation of CD. METHODS/STUDY POPULATION: We conducted a survey with 267 childcare teachers who had been trained in CD across the state, representing early childhood educational environments in urban and rural settings. Specific questions were asked related to level of CD implementation, perceived benefit, and facilitators/barriers to full implementation. A random subset of the sample (8 teachers) participated in a subsequent focus group to explore survey themes in greater depth. Focus group members were asked about their rationale for attending CD training, CD implementation (including barriers/facilitators to full implementation), and perceived impact on their classrooms. The focus group was recorded and transcribed to capture questions and comments. RESULTS/ANTICIPATED RESULTS: Objective 1: 1. 30% of teachers reported full implementation of CD. 2. 50% of teachers reported partial implementation of CD. Objective 2: 1. The vast majority of teachers (95%) agreed that CD had a positive impact on their classroom, including better structure and enhanced relationships with the children. 2. The vast majority of teachers (85%) agreed that CD had a positive impact on the children in their classroom, including increases in problem-solving abilities and self-control. Objective 3: 1. Most teachers (71%) reported experiencing barriers to CD implementation, with the majority of those surveyed (93%) stating that additional implementation support would be helpful. 2. The top three barriers to implementation elicited in survey and focus groups included uncertainty regarding how to begin implementing CD in the classroom, lacking materials for CD implementation, and lacking time to focus on applying knowledge from training into the classroom. 3. The top three facilitators for implementation elicited in survey and focus groups included coaching support for teachers, training agency leadership in CD, and greater perceived impact of CD. DISCUSSION/SIGNIFICANCE OF IMPACT: Childhood disruptive behaviors are among the most frequent reasons for referral to specialized services in and out of the classroom (Sukhodolsky, Smith, McCauley, Ibrahim, & Piasecka, 2016). Disruptive and aggressive behaviors are problematic, not only for victims of children who are aggressive but also for aggressive children as they age. Although effective treatments exist, the level of effective implementation of these interventions are understudied. These results demonstrate that 2/3 of teachers trained in CD are not fully implementing the model and provides concrete barriers and facilitators to current implementation. These data will provide the initial foundation for the development of a targeted implementation strategy that supports full implementation of CD within early childhood education settings.

OBJECTIVES/GOALS: Many museums and art spaces conduct programming intended to improve health outcomes, but arts professionals’ perceptions of these programs are not well known. This study describes arts professionals’ experiences with museum and art-space interventions intended to improve health. METHODS/STUDY POPULATION: A 14-item digital Qualtrics survey was administered to museums and arts organizations selected using snowball sampling. The survey was sent to a range of arts and cultural organizations and professional membership bodies in the US and UK. Survey questions assessed the range of audiences involved in health programs, what types of activities museums and arts organizations are offering to support health outcomes, and how programs are evaluated. RESULTS/ANTICIPATED RESULTS: From 10/30/19-11/19/19, 151 surveys were completed; 66 respondents (44%) have a museum/arts in health program. Common target populations include individuals with mental health concerns (33, 22%) and older adults (26, 17%). Improving wellbeing (56, 37%) and social isolation (50, 33%) were the most common intended outcomes. Respondents reported using a variety of program evaluation methods including formal (23, 15%), informal (31, 21%), and anecdotal (37, 25%). Interviews are planned with a purposive sample of respondents conducting, or interested in conducting, a program for individuals with chronic pain and those formally evaluating their programs. DISCUSSION/SIGNIFICANCE OF IMPACT: Interim survey responses indicate many cultural organizations engage in programming intended to improve health outcomes. Understanding the cultural sector’s current efforts to improve health represents an initial step in translating these efforts into effective intersectoral research partnerships.

OBJECTIVES/GOALS: To describe cognitive and psychiatric symptom profiles of individuals with bvFTD and hyperorality. We test two hypotheses: (1) individuals with hyperorality show more severe psychiatric profiles and (2) neuroanatomic correlates of hyperorality in advanced bvFTD differ from those with early bvFTD. METHODS/STUDY POPULATION: Participants were enrolled in ALLFTD–a multi-site longitudinal study in FTD. We selected the 354 participants who had a primary clinical diagnosis of bvFTD, 344 of whom had data on hyperorality. Each participant underwent extensive clinical interviews and examinations, structural neuroimaging, and blood sampling. Five anatomic regions of interest were identified and analyzed based on previously identified neuroanatomic correlates of hyperorality. Differences in participant characteristics and clinical outcomes were compared using t-tests for continuous variables and Pearsonχ2 tests for categorical variables. Linear multivariate regression controlling for age and total intracranial volume (TIV) was used to examine associations between atrophy in regions of interest and hyperorality status. RESULTS/ANTICIPATED RESULTS: Early-stage participants with hyperorality had poorer self-monitoring, empathic concern, and perspective taking as well as higher CDR behavioral subscale scores compared to those without hyperorality. Advanced stage participants with hyperorality had higher scores on the Social Behavior Observer Checklist compared to those without hyperorality. Early-stage participants with hyperorality displayed higher rates of ritualistic/compulsive behavior and motor disturbance. Advanced stage participants had higher rates of apathy, ritualistic/compulsive behavior, anxiety, and elation. In the advanced stage participants, hyperorality was associated with atrophy in the right dorsal striatum, the right ventral striatum, and the right insula cortex. DISCUSSION/SIGNIFICANCE: Hyperorality emerges early and is accompanied by neuropsychiatric symptoms prior to significant neurodegeneration. Overtime, participants with hyperorality develop more psychiatric symptoms as well as atrophy in striatal and insular brain regions. Our findings suggest a role for novel interventions like non-invasive brain stimulation.

OBJECTIVES/GOALS: Inflammatory Breast Cancer (IBC) is the most aggressive form of breast cancer and does not have targeted therapy. GPR30, a 7-transmembrane estrogen receptor, may play a role in regulating cell growth and proliferation of cancerous cells. Here, we evaluated changes in gene expression while inhibiting GPR30 to determine putative targets to treat IBC. METHODS/STUDY POPULATION: IBC cell lines (SUM149PT) were cultured in medium with serum stripped from growth factor and hormones for 48 hours. Cells were then exposed to either G15 (GPR30 inhibitor) at a concentration of 1µM or ETOH (vehicle negative control) 3 hours in triplicates. After exposure, total RNA was extracted using the Qiagen RNAeasy Mini kit and RNA was sequenced using the Illumina NextSeq (2 X 75bp). The higher-quality reads were aligned, annotated, and quantified to the human genome (HG38) using STAR and RSEM softwares. Gene expression analysis was performed in R statistical software (packages tximport and DESeq2). Functional and enrichment analyses were performed using Metascape and STRING database, respectively. RESULTS/ANTICIPATED RESULTS: There were 656 significantly expressed genes (p < 0.05) between groups (G15 vs. ETOH). The top 5 significant genes include: SMIM7, FANCG, ARID1A, MAML2, and ATF3. Significantly impacted biological processes and pathways include: electron transport chain, mitotic cell cycle process, microtubule cytoskeleton organization, cellular component morphogenesis and DNA-dependent DNA replication (adj p < 0.05). Additionally, physical and functional interaction networks showed 3 major clusters (≥ 12 genes), which contained several gene hubs including BRCA1, BRCA2, FOS (proto-oncogene), PLK1 and PAK1 (both serine/threonine-protein kinases), among others. Interestingly, the network analysis showed the previously known interaction between FANCG and BRCA2, which were both dysregulated by GPR30 inhibition. DISCUSSION/SIGNIFICANCE: Through gene expression, functional and enrichment analyses we found several targets genes that could be associated with the pathogenesis of IBC. Validation of candidates genes (qRT-PCR and Western blot), and functional assays (cell proliferation, motility, and invasion) will be performed to understand the potential of these genes in treating IBC.

OBJECTIVES/GOALS: The goal of this study is to determine the role of the tRNA modifications in the translation of Hfq. Hfq is an RNA chaperone that acts as a co-factor for the action of the largest class of small RNAs in E. coli. RNA modifications have been known to play critical roles in the translational fidelity of many cellular proteins in bacteria. METHODS/STUDY POPULATION: In this study, we used an hfq-lacZ translation fusion to screen several RNA modification mutant genes to uncover additional RNA modifications that may play a role in Hfq translation. We measured hfq-lacZ activity in genetic backgrounds mutated for several additional RNA modification enzymes previously untested for Hfq effects. RESULTS/ANTICIPATED RESULTS: We identified 5 RNA modification genes that were defective for hfq-lacZ fusion activity, and we subsequently performed western blot analysis on the Hfq protein in the absence of these modification mutant genes to determine the effect of these mutants more directly on Hfq protein levels. We identified 2 out of these 5 RNA modification mutants that also affect Hfq protein levels. DISCUSSION/SIGNIFICANCE: Since Hfq is critically important for small RNA function is a wide range of bacteria, it is possible tRNA modifications regulate Hfq expression in other bacteria. These processes, when further investigated, could provide us with the basic information to develop new antibiotics needed to address emerging antibiotic resistance.

ABSTRACT IMPACT: We present preliminary data and an outlined approach to assess peripheral immune changes associated with PTSD in a clinical setting and in a pre-clinical rat model of PTSD. OBJECTIVES/GOALS: We report our methodology and findings indicating a relationship between CNS dopamine signaling and peripheral immune cell populations and propose to extend this methodology to a PTSD patient population to elucidate immune-brain connections in this disorder. METHODS/STUDY POPULATION: Using an IRB-approved protocol in collaboration with a board-certified psychiatrist, we will recruit PTSD patients undergoing treatment, newly diagnosed drug-naiive PTSD patients, and age-matched healthy controls. Flow cytometry will be used for immunophenotyping on blood samples from each group. To complement this data, we will also measure serum cytokine levels in each group. In order to elucidate the connection between the observed immunophenotypes in the PTSD population and CNS neurotransmitters levels, we will employ a rodent model of PTSD and high-pressure liquid chromatography to measure dopamine levels in tandem with peripheral immune changes. RESULTS/ANTICIPATED RESULTS: In both humans and rodents with low CNS dopamine, an expansion of monocyte-derived suppressor cells was observed via flow cytometry. We anticipate that human PTSD patients will exhibit a similar expansion in suppressive immune cells’‘ in agreement with existing literature suggesting a chronic inflammatory state in PTSD. Moreover, in an animal model of PTSD we anticipate an inverse correlation between the CNS dopamine levels and the size of the immune suppressor cell population. DISCUSSION/SIGNIFICANCE OF FINDINGS: Our findings will indicate whether altered dopamine neurotransmission underlies peripheral immune system changes in the context of PTSD models and human patients. Thus, these findings will provide an alternative avenue for future investigations on the role of the immune system in PTSD.

OBJECTIVES/SPECIFIC AIMS: To examine rural-urban disparities in prevalence of diagnosed diabetes in veterans receiving care at the VA and to determine the extent to which demographic factors and obesity levels contribute to identified disparities. METHODS/STUDY POPULATION: A retrospective serial cross-sectional analysis was employed. A stratified weighted random sample of veterans who received care at a VA facility was selected each year for 2007 through 2012. Rural Urban Commuting Area (RUCA) codes were based on resident zip code. Diabetes was defined by two or more primary or secondary ICD-9 codes for diabetes (250.xx) within a 12 month period. Data were analyzed using complex survey-specific procedures. RESULTS/ANTICIPATED RESULTS: Diabetes prevalence 2007-2012 was lowest in urban (20.5%-21.0%), followed by highly rural (21.1%-22.1%) and rural (22.3%-23.0%) areas with the prevalence being significantly higher on the insular islands (31.0%-32.4%). In 2012, 41% of urban, 43% of rural and highly rural and 30% of insular island veterans were obese. Relative to urban areas, the odds ratio for prevalent diabetes was 1.10 (95% CI: 1.08, 1.12) for rural veterans, 1.19 (95% CI: 1.16, 1.23) for insular island veterans, and 1.00 (95% CI: 0.98, 1.02) for highly rural veterans. DISCUSSION/SIGNIFICANCE OF IMPACT: Prevalence of diagnosed diabetes is high in veterans residing in rural, highly rural and urban areas, but markedly higher on the insular islands. Understanding the burden of disease and factors driving disparities provides information required to develop targeted interventions.

OBJECTIVES/GOALS: Participants will be able to identify how partnering with state health departments for a CTSA initiative assists in meeting shared goals by building infrastructure and sharing resources and list key components of a successful county engagement process that can be replicated. METHODS/STUDY POPULATION: Connections IN Health is a collaborative project with the Indiana Clinical and Translational Sciences Institute, Indiana Department of Health, Indiana University Simon Comprehensive Cancer Center, and community partners. We address chronic disease in Indiana with coalition-based engagement at the grassroots level, and partnering to provide technical assistance, resources, connections and evidence-based strategies to address the health challenges. Our methods of county engagement include surveying the broad community, conducting listening sessions with key community stakeholders, compiling, analyzing and sharing data, collaborating with existing local coalitions, assessing community readiness and suggesting evidence-based practices to implement. RESULTS/ANTICIPATED RESULTS: As a result of county engagement, local coalitions have current, local data to drive their efforts in improving local health rankings and outcomes. Community partners in eight counties have selected and implemented evidence-based strategies to manage and/or prevent diabetes, cardiovascular disease and/or stroke. Some examples include: starting a community garden with cooking classes in a food desert to provide fresh produce and nutrition/cooking knowledge, creating a mobile kitchen on hand for organizations to use to provide education and cooking skills, providing a passport for wellness at existing community events with food samples and health screenings and offering evidence-based educational programming such as Dining with Diabetes and Be Heart Smart. DISCUSSION/SIGNIFICANCE: Moving county health coalitions to evidence-based programming that has the greatest likelihood for success is a critical translational sciences challenge. Our rigorous and well defined approach yields significant improvement in local health coalition activities, sustaining their activities through long-term trusted relationships.

OBJECTIVES/SPECIFIC AIMS: To study the role functional capacity plays in surgical outcomes for head and neck cancers. METHODS/STUDY POPULATION: In this single-institution cohort study, we combined preoperative anesthesia assessment information with oncology registry data for newly-diagnosed patients with squamous cell carcinoma of the oral cavity, pharynx, and larynx (HNSCC) treated with definitive surgery at Siteman Cancer Center from 2012 to 2016. Patient-reported exercise capacity was assessed as metabolic equivalents. Metabolic equivalents<4 was defined as poor functional capacity. The primary outcome measure was overall survival (OS). Kaplan-Meir survival analysis was used to compare the survival of patients with poor functional capacity (PFC) and patients with normal functional capacity (NFC). Cox proportional hazard regression was used to explore the independent prognostic role of functional capacity on overall survival after controlling for other factors. RESULTS/ANTICIPATED RESULTS: A total of 671 patients underwent surgical treatment for HNSCC. The average age was 62 years (range: 19–94 years). Majority of the patients were male (n=481; 72%), White race (n=589; 88%), and smokers (n=528; 79%). Of 671 patients, 22% (n=146) had PFC. Two-year OS rate in PFC patients was 70% compared with 85% in NFC patients (15% difference; 95% CI: 7%–23%). Unadjusted Cox proportional hazard analysis showed that PFC patients had 2.2 times higher risk of death (95% CI: 1.5–3.2) than NFC patients. After adjustment for age at surgery, BMI, preoperative weight loss, comorbidity score, tumor site, and TNM stage the magnitude of the association between functional capacity and OS decreased (aHR=1.3; 95% CI: 0.88–1.98). DISCUSSION/SIGNIFICANCE OF IMPACT: Poor functional capacity is associated with decreased overall survival, but the magnitude of the association, while clinically meaningful, decreases after controlling for other important patient and tumor factors. Nevertheless, we believe preoperative functional capacity status is an important patient factor to consider when discussing prognosis and attempting risk stratification. We also believe that functional capacity may be associated with 30-day unplanned readmissions and 90-day complications and are currently performing chart review to ascertain this information.

OBJECTIVES/GOALS: Translational Science includes the social responsibility to improve health outcomes and reduce disparities. This study aims to determine how scientists and community members perceive the social responsibility of translational science. METHODS/STUDY POPULATION: Methods include a series of 12 one-hour semi-structured focus groups. Six groups were held with a total of 34 scientists and four with approximately 32 community members (community focus groups still ongoing). Sample includes scientists and community members affiliated to one of three research fields: HIV/AIDS, Maternal Health, and Mental Health/Substance Use. Inclusion criteria for scientists were based on their scholarly contributions to a field. Community members included individuals who self-identified as affiliated to any above fields. Using reflexive thematic analysis, we analyzed the transcripts against existing frameworks on social responsibility. RESULTS/ANTICIPATED RESULTS: Literature on social responsibility suggests that translational science should be relevant, usable and sustainable for end-users. Scientists and community members identified the same groups for whom research is relevant. Scientists often conceptualized social responsibility through the sustainability lens, focusing on results that might have long-term societal impacts. Due to this, they saw limitations to social responsibility in academia and funding structures. Community members often conceptualized social responsibility through the usability lens, focusing on the complexity of research use and access. They saw barriers to social responsibility in systemic disparities. DISCUSSION/SIGNIFICANCE: Although participants emphasized different practices in focus groups, participants seemed generally interested in pursuing future discussions to develop best practices and commitments regarding social responsibility. Further discussions will take place in a deliberative dialogue activity in January 2024.

OBJECTIVES/SPECIFIC AIMS: Infants with single ventricle congenital heart disease (CHD) who undergo staged surgical reconstruction are among the pediatric patients at highest risk for thrombotic complications. Despite improvements in survival due to medical and surgical advancements, thrombotic complications are common and lead to increased morbidity and mortality, especially during the first two stages of surgical reconstruction. The burden of disease caused by thrombosis is not fully known, and the risk factors associated with thrombosis are not clear. Due to this knowledge gap, prevention of thrombosis with medication, a strategy called thromboprophylaxis, has not been standardized, leading to inadequate prevention of thrombosis. In order to understand the burden of thrombosis and then provide targeted thromboprophylaxis for thrombosis prevention, better characterization of thrombotic complications and the associated factors is needed. Hypothesis: I hypothesize that in infants with single ventricle CHD, the incidence of thrombosis will be more frequent after stage I Versus stage II reconstruction, despite the type of shunt used. Specific demographic, clinical, and surgical variables will be associated with an increased risk for thrombotic complications, and a model to predict which subset of infants is at increased risk will be developed. Specific Aim 1: Characterize the incidence of thrombotic complications at different time points from stage I through stage II of the single ventricle reconstruction (SVR) pathway and determine the demographic, clinical, and surgical factors associated with thrombosis in infants with single ventricle CHD. (1) Determine the incidence of thrombosis in infants with single ventricle CHD. (2) Compare the rate of thrombotic complications between the 2 most commonly used approaches for stage I reconstruction for the group of patients with hypoplastic left heart type of anatomy [modified Blalock-Taussig shunt (MBTS) vs. right ventricle to pulmonary artery shunt (RVPAS)]. (3) Determine the factors (demographic, clinical, and surgical) associated with thrombosis in infants with single ventricle CHD. Specific Aim 2: Determine which subset of infants with single ventricle CHD is at increased risk of developing thrombotic complications across the first 2 stages of surgical reconstruction. (1) Test the identified demographic, clinical, and surgical variables including, but not limited to, gestational age, sex, CHD diagnosis, baseline oxygen saturation, stage of reconstruction, shunt type, and other clinical data available in a univariable and multivariable analysis and study their potential interactions to construct a novel risk predictive model specific for single ventricle CHD. METHODS/STUDY POPULATION: To address the specific aims, I will utilize data from the SVR clinical trial public use data set. This data set includes a prospective cohort of infants, 0–14 months of age, enrolled from any of the 15 participating clinical centers from the years 2005 to 2009. Inclusion criteria for enrollment were diagnosis of hypoplastic left heart syndrome or related single, morphologic right systemic ventricle anomaly, planned Norwood procedure, and informed consent of parent or legal guardian. No additional subjects outside of this data set will be included. Exclusion criteria were a diagnosis of single, morphologic left ventricle anomaly, preoperative identification of anatomy rendering the MBTS or RVPAS technically impossible, and any other major abnormality or acquired extra-cardiac disorder that could independently affect the likelihood of the subject meeting the primary endpoint. The complication of stroke will be excluded from the analyses of factors associated with thrombosis. The complication of thrombosis as defined in this dataset is a composite of events that include arterial or venous thrombosis, thromboembolism, and pulmonary embolism. The data was collected in such a way that it will not be possible to separate these sub-types of thrombosis. Additional thrombotic events of interest are superior vena cava occlusion and inferior vena cava occlusion. Specific Aim 1: Patient data will be extracted from the SVR clinical trial public use dataset to characterize the incidence of thrombotic complications at different time points from stage I through stage II of the SVR pathway and determine the demographic, clinical, and surgical factors associated with thrombosis in infants with single ventricle CHD. In addition, I will compare the rates of thrombotic complications between the 2 most commonly used approaches for stage I palliation for the group of patients with hypoplastic left heart type of anatomy (MBTS vs. RVPAS) and will test the hypothesis that the risk of thrombotic complications is associated with the stage of palliative surgery (stage I vs. stage II). Specific Aim 2: We will test identified demographic, clinical, surgical, and newly identified variables in a univariable and multivariable analysis and study their potential interactions to construct a novel risk predictive model specific for single ventricle CHD. RESULTS/ANTICIPATED RESULTS: To determine feasibility for adequate numbers to be able to address the research aims, a preliminary analysis dataset was performed using a dataset from the Pediatric Heart Network. The PHN is a collaborative group of hospitals that participates in clinical research studies in children with CHD. For the SVR clinical trial, the PHN conducted a randomized clinical trial at 15 centers in North America between 2005 and 2009, prospectively enrolling infants with HLHS or single right ventricle anomalies who were to undergo the Stage I Norwood procedure. A total of 920 newborns were screened; 664 were medically eligible and 549 patients were randomized. The primary aim of the trial was to compare survival of infants randomized to receive either the Norwood procedure with the MBTS or the RVPAS. These patients were followed at specific time points, including from baseline (pre-Norwood), at the time of the Norwood procedure, between stage I and II, following stage II reconstruction, and at 14 months of age. At these time points, data were collected that includes demographic, radiologic, clinical, and surgical outcomes. Included in the clinical outcomes are complications, such a thrombosis. There was no screening process to assess for asymptomatic thromboses, suggesting that most, if not all, discovered thromboses were due to clinically relevant effects. A newer iteration of this study (SVRIII) expands the monitoring of this cohort until the Fontan stage at 2–6 years of age, but these data have not yet been released in the public use data set. A descriptive analysis of the frequency of thrombotic complications was assessed at each time point, as well as in aggregate. Data were extracted from the specific time periods of interest, identified as Pre-Norwood, during Norwood Hospitalization, in-between visits, and during Stage II Hospitalization. There were 549 infants who were randomized with available data to analyze. During the Norwood hospitalization, 37 infants had a thrombotic complication. Between Stage I and Stage II outpatient visits, 8 infants had a thrombotic complication. During Stage II hospitalization, 16 infants had a thrombus. Overall, 61 individual patients (11%) had a thrombotic complication. DISCUSSION/SIGNIFICANCE OF IMPACT: This study utilizing data from the Pediatric Heart will be the largest cohort ever utilized for characterizing thrombotic complications and determining the factors associated with thrombosis across the first and second stages of surgical reconstruction. More than 500 (n=549) subject’s data will be analyzed through the first two stages of reconstruction, while the largest analysis before this proposed analysis only included a total of 195 children. Notably, these prior studies did not include a comparison between the 2 shunt types in stage I reconstruction, leaving a gap in knowledge regarding the incidence of thrombosis comparing these groups. The analysis will be the first to address this gap and update the current literature. Preliminary data show that the overall incidence of thrombosis across the first 2 stages of surgical reconstruction was 11%, which is lower than the previously reported overall rates of 40%–50%. Despite the continued lack of evidence-based guidelines for thromboprophylaxis methods, the decreased overall rate is most likely due to more widespread practice of anticoagulation in general. Determining the factors associated with thrombosis across the first and second stages of surgical reconstruction will help identify those at risk. An innovative aspect of this analysis will be the use of disease-specific factors to develop a model to predict thrombosis. Unique factors include cardiac variables like ejection fraction, baseline oxygen saturation, shunt type (MBTS vs. RVPAS), and other echocardiographic parameters. While the use of thromboprophylaxis has been associated with decreased risk of thrombosis, there is no general consensus to guide thromboprophylaxis in this population, which can be burdensome and costly. Determining which subset of infants with single ventricle CHD are at increased risk of developing thrombotic complications will allow for the development of a prediction model to predict those at highest risk of developing a thrombotic complication. Developing a predictive model will be a novel way to identify patients at risk for thrombosis and will set the stage for targeted prevention of thrombosis.

OBJECTIVES/GOALS: This research project aims to isolate and identify bioprospects capable of metabolizing estrogen using culture-dependent, culture independent methods and the identification of the gene/genes responsible for the metabolization of estrogen by the bioprospects. METHODS/STUDY POPULATION: For the culture dependent technique, samples were collected from the water treatment plant in Mayagüez, cultivated on TSA medium and selected specific and diverse colonies were patched on M-9NC (no carbon sources), M-9-glucose (M9G) and M-9-hormone mixture (M9H: 17-beta-Estradiol and 17-alpha-Ethynylestradiol). After the 48hrs incubation at 25 and 37 Celsius, growth was scored on the different media, to choose those potential bioprospects that use the hormones as the sole carbon source. For the culture independent approach, metagenomic clones from libraries generated from the Guajatacas water reservoir and Cabo Rojos Saltern microbial mats were cultivated on the appropriate M9NC, M-9G and M-9H, supplemented with Leucine and Thiamine for 48 hours at 37 Celsius. RESULTS/ANTICIPATED RESULTS: The culture-dependent approach showed a total of 6 potential bioprospects capable of growing on hormones as a sole carbon source on M9H at 25 and 37 Celsius and no-growth on the M-9 media used. Currently, we are in the process of determining the identities of the cultivable isolates genetically, any preference on hormone metabolization, monitoring a larger number of metagenomic clones and optimizing the selection conditions. DISCUSSION/SIGNIFICANCE: The identification of bioprospects capable of metabolizing hormones could potentially lead to the generation of new water treatments that could help remove unwanted chemicals, such as high concentrations of estrogens.

OBJECTIVES/GOALS: While there are many well-documented factors for racial/ethnic variation in discharge destination, less is known about the role hospital processes play. We hypothesize that variation in hospital processes -defined as the patient length of stay (LOS) adjusted for known confounders - explains racial/ethnic variation in discharge destination. METHODS/STUDY POPULATION: Our sample was 176,686 discharges from 165 hospitals in 2014 using the New York State Inpatient Dataset from the Healthcare Cost and Utilization Project, merged with the 2014 American Hospital Association Annual Survey to build a file of inpatient discharges with patient, disease and socio-economic characteristics. We excluded patients under 18 years, those with LOS of zero, those who died, those admitted to critical access hospitals, and patients from hospitals that lacked sufficient number of minority patients. We used a generalized linear mixed effects model to create an in-hospital risk-adjusted LOS by modelling the relationship between the interaction of race and discharge destination and LOS, controlling for known confounders such as patient, disease and between-hospital characteristics. RESULTS/ANTICIPATED RESULTS: Mean age of sample was 56.5 years, 57.3 % female, 54.9% white, 18.9% black and 13.1% Hispanic; 64.3% were discharged home, 15.8% to a skilled nursing or other intermediate care facility, 15.5% to home with home care and 2.4% left against medical advice. The top 3 discharge diagnoses were vaginal delivery (6.3% of discharges), psychosis (4.7%), and major joint replacement (2.9%). In adjusted analysis compared to white patients, black and Hispanic patients did not have an risk of increased LOS after being discharged to non-home destinations vs. discharged home, (black patients, adjusted OR [AOR], 0.97; 95% CI: 0.94-1.00, p = 0.08; Hispanic patients, AOR, 1.01; 95% CI: 0.98 – 1.05, p = 0.5). However, being black compared to white and discharge to non-home destinations significantly increased LOS. DISCUSSION/SIGNIFICANCE OF IMPACT: In this large sample of patients admitted for inpatient care in 2014 in New York, we found no independent effect between race and discharge destination on a patient’s LOS after controlling for patient, disease and between-hospital characteristics. However race/ethnicity increased LOS, suggesting its effect may play a role on in-hospital processes CONFLICT OF INTEREST DESCRIPTION: Dr. Ghosh has no relevant relationships with commercial interests to disclose Dr. Ibrahim has no relevant relationships with commercial interests to disclose

OBJECTIVES/GOALS: Our objective is to establish a proteomic protein labeling method from tumor tissue and blood samples obtained from patients undergoing surgery for pancreatic cancer in Puerto Rico. Our goal is to discover potential biomarkers in the patient tumor/blood samples that are not expressed in normal control samples obtained from potential organ donors. METHODS/STUDY POPULATION: A pilot study with ten patients undergoing surgery for pancreatic cancer will obtain tumor tissue and blood samples. Protein extracts isolated from tissue/cells will be reduced, alkylated, and digested overnight. Samples will be labeled with TMT reagents and mixed before fractionation and cleanup. Labeled samples will be analyzed with a high-resolution Orbitrap LC-MS/MS before data analysis to identify peptides and quantify the reporter ions. The altered proteins will be analyzed by ELISA to confirm their presence. The protein arrangements will be compared with results from proteomic profile banks to assess their prevalence. As controls, parallel protein analyses will be performed on normal tissue/blood samples from organ donors, facilitated by our local organ procurement organization. RESULTS/ANTICIPATED RESULTS: We anticipate finding proteogenomic material defining PC and new proteomic subtypes not previously described in this population. In addition, studying protein overexpression and underexpression can identify relevant genes and potential biomarkers. We hypothesize that PC in the Hispanic population will show slight variations in tumor protein expression than in other populations, which could lead to the discovery of a new Hispanic-specific biomarker. DISCUSSION/SIGNIFICANCE: We expect to provide essential information that will influence the next steps in developing future screening tests. Identifying specific proteins with the potential to become a preventive test should eventually lead to a reduction in morbidity and mortality of PC. The results of this work should lay the foundation that can guide future research.

OBJECTIVES/GOALS: This study utilizes a Black Feminist metatheoretical approach in conjunction with the Social Amplification of Risk Framework to: i) assess the role of communication with clinicians on Black women’s perceptions of risk and ii) outline recommendations for empowering communication that attenuates Black women’s perceptions of pregnancy risk. METHODS/STUDY POPULATION: Semi-structured in-depth interviews were conducted with 28 self-identified Black women in 2022 and 2023 to gain insights into their conceptualizations of pregnancy risk, and the role of communication with clinicians in the amplification and/or attenuation of pregnancy risk perceptions.An IRB approved flier was used for both in-person and online recruitment of participants from 12 US states.Each participant signed a consent form and received $20 as compensation for their time. Interviews were recorded and transcribed using a digital transcription tool, and then reviewed and edited to ensure accuracy. This study’s metatheoretical underpinning in Black Feminist Theory informed the use of reflexive thematic analysis in the qualitative analysis of the in-depth interviews. RESULTS/ANTICIPATED RESULTS: This study found thatunclear communication (including insufficiently contextualized history taking), obstetric violence, anddiagnoses with insufficient explanation,led to the amplification of concerns and worries about pregnancy as well as erosion of trust in healthcare. In contrast,clinicians lessenedconcerns about pregnancy through clear communication (including appropriately contextualized history-taking), demonstration of empathy, and by fostering patient agency.The study thus highlights the importance of clinicians' clear communication,demonstration ofempathy and fostering of patient agencyas critical factors to empowering communication that attenuates Black women’s pregnancy-related risk perceptions. DISCUSSION/SIGNIFICANCE: Given the unjust dangers Black women face during pregnancy, this study demonstrates how patient-clinician communication influences Black women’s pregnancy risk perceptions, providing recommendations for clinician communication practices that empower Black women and attenuate their perceptions of pregnancy risk.

OBJECTIVES/SPECIFIC AIMS: The primary objective of this study was to assess the interest in using mobile technology to manage hypertension and assessing medication adherence among African Americans and Hispanics living in a low-income urban community in Central PA. METHODS/STUDY POPULATION: This qualitative research study was designed using structured interviews with 30 African American participants with hypertension. The study survey consisted of 43 questions which included demographics, use of technology, technology literacy, health literacy, and medication adherence. Participants self-reported their responses about use of technology, literacy, and adherence. The data was examined using summary statistics. RESULTS/ANTICIPATED RESULTS: Out of 30 participants, 61% identified as African American and 30% identified as Hispanic. Overall, participants reported high use of technology; with 63.3% using the internet occasionally, 73.3% having a cell phone, and 66.7% accessing the internet using a mobile handheld device. Participants expressed an interest in measuring blood pressure, accessing health information, and managing medication on their cellphone or tablet. Twenty-five percent of participants self-reported not being confident at all in managing their medication without help and remembering to take their medication as prescribed. Fewer participants (54.2%) were very confident with using technology to help manage medication and treatment. DISCUSSION/SIGNIFICANCE OF IMPACT: This study provides important insights on the use of technology for managing hypertension in an urban low-income community. Results suggest that medication adherence, the use of technology, and the willingness to use mobile applications for hypertension management is high within this low-income community. This study gives future researchers a framework on how to assess these specific areas in their own population.