OBJECTIVES/GOALS: Academic research centers often struggle to recruit and retain a diverse and competent clinical and translational science (CTS) workforce. Specifically, the clinical research professional (CRP) career pathway is not well known to undergraduate students and other individuals outside of academic medicine despite various potential career routes. METHODS/STUDY POPULATION: To address these workforce challenges, the CRP Task Force at the University of Cincinnati (UC) aims to train a competent and diverse CRP workforce through targeted educational programming in the UC undergraduate population. Using a six-step curriculum development process that included: 1) performing a needs assessment, 2) determining content, 3) writing goals and objectives, 4) selecting the educational strategies, 5) implementing the curriculum, and 6) evaluating the curriculum, we designed an undergraduate certificate program in CTS. RESULTS/ANTICIPATED RESULTS: The needs assessment included both internal and external data gathering to inform curriculum development and program decisions. Content was determined using the CRP Competency Framework 2.0, and program learning outcomes were written with both the competency framework and local workforce needs in mind. Educational strategies were selected based upon optimization of available resources and local expertise with an emphasis on interactive didactics complemented by experiential learning. Implementation is underway and evaluation will follow once students begin enrolling. DISCUSSION/SIGNIFICANCE: We anticipate an increase in numbers of well-qualified, diverse applicants who pursue CRP careers locally and regionally. In addition, we expect that the certificate program will build competency earlier in CRP staff, improving job satisfaction and retention as a result of a stronger foundation from which to build their professional skills.

OBJECTIVES/GOALS: Young and middle-aged adults with a myocardial infarction (MI) represent an understudied group potentially with unique risk indicators such as emotional stress. We sought to investigate if mental stress-induced myocardial ischemia (MSIMI), a marker of cardiovascular vulnerability to psychological stress, is associated with poor outcomes among this population. METHODS/STUDY POPULATION: We studied 306 patients (150 women and 156 men) ≤61 years of age who were hospitalized for MI in the previous 8 months. Clinical, behavioral and psychosocial factors were assessed with standardized measures. Patients underwent myocardial perfusion imaging with mental stress (public speaking) and conventional stress (exercise or pharmacological testing). MSIMI and conventional stress-induced ischemia were defined as a new or worsening perfusion defect. Patients were followed for 3 years for adverse events, which were independently adjudicated. Cox proportional hazard models were used to estimate the association of MSIMI and CSIMI with a composite endpoint of recurrent MI or cardiovascular (CV) death with adjustment for demographic, clinical and psychosocial risk factors. RESULTS/ANTICIPATED RESULTS: The mean age of the sample was 50 years (range, 22-61). MSIMI occurred in 16% of the patients, and conventional ischemia in 35%. Over a 3-year follow-up, 28 individuals had a recurrent MI and 2 died due to cardiovascular causes. The incidence of the composite endpoint of MI or CV death was more than doubled in patients with MSIMI (20%) than those without MSIMI (8%), HR 2.6, 95%CI, 1.2-5.6. Further adjustment for demographic and clinical risk factors and depressive symptoms did not substantially change the relationship. In contrast, conventional stress ischemia was not significantly related to the outcome (HR 1.4, 95%CI, 0.6-3.0). DISCUSSION/SIGNIFICANCE OF IMPACT: Young and middle-aged individuals with MSIMI after MI have a >2-fold higher likelihood of recurrent MI and CV mortality compared with those without MSIMI. In this patient group, MSIMI is a better risk indicator than ischemia with a conventional stress. These findings point to psychological stress as an important determinant of risk in this patient population. Ischemia induced by mental stress is a potent risk indicator in young post-MI patients. Stress-reduction interventions may be especially beneficial in patients who show this abnormal response.

OBJECTIVES/SPECIFIC AIMS: Systemic sclerosis (SSc) is a connective tissue disease of unknown etiology characterized by progressive fibrosis of the skin and multiple visceral organs. Effective therapies for SSc are needed. Lysyl oxidase (LOX) is a copper-dependent amide oxidase that plays a critical role in the crosslinking of the extracellular matrix (ECM). In this study, we investigated the role of LOX in the pathophysiology of SSc. METHODS/STUDY POPULATION: LOX expression and protein levels were measured in lung tissues and primary fibroblasts from patients with SSc and healthy controls. The effects of recombinant LOX (rLOX) were measured in vitro in primary fibroblasts, ex vivo in human lung tissues and in vivo in mice given bleomycin in combination with rLOX. LOX levels and activity were evaluated in lung fibroblasts treated with an endostatin-derived peptide that ameliorates fibrosis and in mice treated with bleomycin in combination with the peptide. Further, to differentiate the crosslinking activity of LOX from other potential effects, primary human fibroblasts were cultured with rLOX in the presence of the inhibitor, beta-aminopropionitrile. The expression levels of ECM (collagen and fibronectin), pro-fibrotic factors (IL-6 and TGF-beta), and transcription factor (c-Fos) were examined by real-time PCR, ELISA, immunoblotting, or hydroxyproline assay. RESULTS/ANTICIPATED RESULTS: LOX mRNA was increased in lung tissues and matching fibroblasts of SSc patients. rLOX-induced ECM production in vitro and ex vivo in lung fibroblasts and in human lung tissues maintained in organ culture, respectively. Additionally, TGF-beta and bleomycin induced ECM production, LOX mRNA expression and activity. Endostatin peptide abrogated these effects. In vivo, rLOX synergistically exacerbated pulmonary fibrosis in bleomycin-treated mice. The inhibition of LOX catalytic activity by beta-aminopropionitrile failed to abrogate LOX-induced ECM production. LOX increased the production of IL-6. IL-6 neutralization blocked the effects of LOX. Further, LOX induced c-Fos expression and its nuclear localization. DISCUSSION/SIGNIFICANCE OF IMPACT: LOX expression and activity were increased with fibrosis in vitro, ex vivo, and in vivo. LOX induced fibrosis via increasing ECM, IL-6 and c-Fos translocation to the nucleus. These effects were independent of the crosslinking activity of LOX and mediated by IL-6. Our findings suggest that inhibition of LOX may be a viable option for the treatment of lung fibrosis. Further, the use of human lung in organ culture establishes the relevance of our findings to human disease.

OBJECTIVES/GOALS: Murine and clinical data suggest that antibody-dependent cellular cytotoxicity (ADCC) is associated with greater protection against disseminated neonatal HSV disease. To quantify the relative transfer of Abs with different functions and targets, we conducted a prospective study of mother-infant term and preterm dyads pre and during COVID-19 METHODS/STUDY POPULATION: Total and HSV lysate, glycoprotein D (gD) and glycoprotein B (gB)-specific IgG, IgG1 and IgG3 as well as HSV neutralizing Abs (nAbs) and ADCC were quantified in paired 3rd-trimester pregnant women and their newborns (cord) blood. Transfer ratios (TR) were defined as cord:maternal Ab levels. IgG1 and IgG3 subclass and gD or gB-specific Abs were isolated by column purification and glycan profiles were assessed by mass spectrometry. The study population included 21 term and 15 preterm dyads who were HSV-1 (+/- HSV-2) seropositive enrolled between 2018-2019 (pre-COVID) and 25 additional HSV-1 (+/- HSV-2) seropositive term dyads whose mothers were SARS-CoV-2 PCR and COVID Ab+ at delivery; 14 were asymptomatic and 11 had mild-moderate COVID disease. None of the mothers had active genital HSV lesions during delivery RESULTS/ANTICIPATED RESULTS: Anti-HSV IgG, IgG1 and IgG3 TR were higher in term vs. preterm dyads (p<0.05). The nAb TR was 2.4 in term vs. 0.8 in preterm (p<0.001) but the ADCC TR was < 1.0 for both. To determine if the latter reflected antigenic target, subclass or glycans, we enriched for gD and gB specific and IgG1 and IgG3 Abs. The gD Abs were IgG1 and had only neutralizing activity. In contrast, gB Abs were polyfunctional and included IgG1 and IgG3 but only the IgG1 Abs had ADCC activity. The gD Abs were enriched for glycans associated with an affinity for the neonatal Fc receptor (FcRn); gB Abs expressed glycans associated with both FcRn and FcγRIIIa binding. There was no significant difference in total HSV-specific IgG TR in pre-COVID vs post-COVID dyads but the nAb TR was lower (p=0.018) and ADCC TR higher (p<0.001) in the COVID compared to pre-COVID cohort DISCUSSION/SIGNIFICANCE: HSV ADCC Abs, which may provide greater protection than nAbs against neonatal disease, transfer poorly particularly to preterm newborns. However, in the setting of SARS-CoV-2, the TR of HSV ADCC is significantly higher. This may reflect alterations in the placental architecture and/or glycan composition which is currently being investigated.

OBJECTIVES/GOALS: To describe how the UCLA Clinical and Translational Science Institute (CTSI) assembled and deployed a science team in support of a local jurisdictions effort to manage and control COVID-19 outbreaks in one of the nations largest metropolitan regions, Los Angeles County (LAC). METHODS/STUDY POPULATION: During the COVID-19 pandemic (2020-21), building an efficient data infrastructure to support outbreak management became a priority for the local health department. In response, the UCLA CTSI assembled a science team with expertise across the translational continuum: epidemiology, laboratory and microbiology, machine learning, health policy, medicine and clinical care, and community engagement. The team partnered with a new LAC Data Science Team to foster a collaborative learning environment for scientists and public health personnel, employing improvement and implementation science to help mitigate COVID-19 outbreaks in sectors including healthcare, skilled nursing facilities, and K-12 education. The goal was a public health workforce that is prepared to problem-solve complex, evolving outbreaks. RESULTS/ANTICIPATED RESULTS: The science team created a learning environment with data modeling and visualization, problem-based learning, and active knowledge and skills acquisition. First, control charts and time series methods were used to visualize COVID-19 data and find signals for action. Second, a series of 16 Grand Rounds offered interactive sessions on problem-solving of outbreak challenges in different sectors. Third, a biweekly Public Health Digest provided fieldworkers with the latest scientific studies on COVID-19. All three elements guided and empowered the workforce to implement timelier, efficient outbreak mitigation strategies in the field. The partnered team also identified barriers to adoption of selected new data and management techniques, revealing areas for further skill-building and data-driven leadership. DISCUSSION/SIGNIFICANCE: The UCLA CTSI science team offered a backbone science infrastructure for helping public health and other sector agencies manage COVID-19 outbreaks and mitigation. It showed promise in bringing and translating science into public health practice. It revealed future priorities for CTSI innovation and scientific support of public agencies.

OBJECTIVES/SPECIFIC AIMS: The current study examined hangover following IV alcohol self-administration (IV-ASA) using the Computer-Assisted Infusion System. The goal of the study was to identify predictors of hangover, including drinking history, alcohol sensitivity, family history, expectancies, and sex differences in nondependent drinkers. METHODS/STUDY POPULATION: The study sample included 89 healthy, nondependent drinkers aged 21–45 years. After a screening to exclude any medical illness or psychiatric disorders, participants completed an IV-ASA session. Each session consisted of a 25-minute priming phase, during which participants were prompted to press a button to receive individually standardized alcohol infusions, followed by a 2-hour “open bar” phase, during which they were instructed to recreate a typical drinking experience. Results from the IV-ASA included peak and average BrAC. Drinking patterns were assessed using the Alcohol Use Disorders Identification Test, which provided 3 subscales: consumption (AUDIT-C), dependence (AUDIT-D), and harmful drinking (AUDIT-H). Subjective response to alcohol was measured using the Drug Effects Questionnaire (DEQ). The Alcohol Hangover Scale (AHS) was used to assess hangover for the period between participants’ departure from the study unit and 10 am the next morning. The Alcohol Effects Questionnaire (AEFQ) is a measure which includes 40 true/false statements about how alcohol typically makes respondents feel, and was used to measure alcohol expectancies. RESULTS/ANTICIPATED RESULTS: Results showed that 78% of participants endorsed having at least 1 hangover symptom following IV-ASA. The most commonly reported items were tired, thirsty, headache, and hangover. There was no association between hangover scores and the AUDIT-C or IV-ASA. Because alcohol consumption was not related to hangover symptoms, risky drinking behavior was examined. Results indicated that participants endorsing 4 or more items on the AUDIT-D plus AUDIT-H subscales showed significantly higher average hangover scores. Linear regression analyses indicated that alcohol hangover scores were associated with DEQ items feel, high, and intoxicated. Ongoing analyses are examining additional predictors of hangover including family history, alcohol expectancies, sex differences, and other alcohol sensitivity measures. DISCUSSION/SIGNIFICANCE OF IMPACT: The results indicated that risky drinking patterns and alcohol response measures were positively associated with hangover symptoms in non-dependent drinks, while no correlation between consumption and hangover symptoms were found. Since previous research has shown than greater subjective response is associated with heavy drinking and predictive of alcohol use disorder, it is possible that hangover symptoms is a marker of this relationship. Since the role of hangover in the transition from heavy drinking to disorder still remains unclear, it will be important to characterize this relationship between alcohol sensitivity and hangover as a function of drinking patterns. This understanding may help to prevent this transition from at-risk drinking to alcohol dependent drinking.

OBJECTIVES/GOALS: Writing self-efficacy & self-regulation are tied to publication & grant submission” writers high in these traits are more productive. We investigated if participating in Shut Up & Write! would produce statistically significant gains in self-efficacy & self-regulation when comparing pre/post surveys. METHODS/STUDY POPULATION: 47 US med students, TL1/KL2, & early-career faculty expressed interest in participating. We conducted a 12-week, 1 hr/week Shut Up & Write!-style (SUAW) activity, held on Zoom, and measured the effect using a pre-post survey adapted from the Writer Self-Perception Scale. Matched pairs t-tests (I2=0.05) to test for significant differences between pre- and post-test means were conducted on 3 subscales. The 3 subscales reflected writing attitudes (self-efficacy), writing strategies (self-regulation), and avoiding writing distractions (self-regulation). Subscales showed acceptable internal consistency with Cronbach’s alphas of 0.80, 0.71, and 0.72. RESULTS/ANTICIPATED RESULTS: 24/37 (65%) completed pre-post surveys. 81% presented as female. 60% were NIH-defined URB and/or were from MSIs. 60% previously participated in an activity similar to SUAW. Sum scale statistics for those who previously participated were significant for the self-efficacy subscale (p=0.020) and writing strategies subscale (p=0.041). Sum scale statistics for those who had not previously participated were significant for the writing strategies subscale (p=0.002). We saw no difference in the avoiding writing distractions subscale. 80% were very satisfied/satisfied with SUAW (I really loved these sessions” they helped me to identify a writing goal that could actually be accomplished in an hour.) DISCUSSION/SIGNIFICANCE: We found significant differences in self-efficacy & self-regulation, building upon findings from a 2021 pilot, and providing evidence that regular participation in an activity like SUAW produces increased self-efficacy and self-regulation and may increase manuscript and grant-writing productivity

OBJECTIVES/SPECIFIC AIMS: Transforming growth factor-beta (TGFβ) is a genetic modifier of cystic fibrosis (CF) lung disease. TGFβ’s pulmonary levels in young CF patients and its mechanism of action in CF are unknown. We examined TGFβ levels in children with CF and investigated responses of human airway epithelial cells (AECs) and mice to TGFβ. METHODS/STUDY POPULATION: TGFβ levels in bronchoalveolar lavage fluid from CF patients (n=15) and non-CF control patients (n=21)<6 years old were determined by ELISA. CF mice and non-CF mice were intratracheally treated with an adenoviral TGFβ1 vector or PBS; lungs were collected for analysis at day 7. Human CF and non-CF AECs were treated with TGFβ or PBS for 24 hours then collected for analysis. RESULTS/ANTICIPATED RESULTS: Young CF patients had higher bronchoalveolar lavage fluid TGFβ than non-CF controls (p=0.03). Mouse lungs exposed to TGFβ demonstrated inflammation, goblet cell hyperplasia, and decreased CFTR expression. CF mice had greater TGFβ-induced lung mechanics abnormalities than controls; both CF human AECs and CF mice showed higher TGFβ induced MAPK and PI3K signaling compared with controls. DISCUSSION/SIGNIFICANCE OF IMPACT: For the first time, we show increased TGFβ levels very early in CF. TGFβ drives CF lung abnormalities in mouse and human models; CF models are more sensitive to TGFβ’s effects. Understanding the role of TGFβ in promoting CF lung disease is critical to developing patient specific treatments.

OBJECTIVES/GOALS: 1. Standardize pathways, training and evaluations 2. Expose apprentices to a variety of research experiences 3. Remove barriers to hiring early talent 4. Expand opportunities for underrepresented minority applicants to obtain clinical research professional positions METHODS/STUDY POPULATION: Collaborators connected by the Clinical Research Professional Taskforce ACTS SIG conducted a landscape analysis survey to identify aspects of CRP Apprentice models and formed a Subgroup. Members will share plans for multiple apprenticeship programs, including specific training modalities and skill sets used to prepare apprentices for a successful clinical research professional career. Methods across institutions include: • Increasing awareness of the profession • Facilitating talent identification for managers • Making the business case for funding and staffing • Implementing work-based learning for fundamental competency development Survey results from CRP institutions demonstrated apprenticeships are value added to teaching how to conduct research. RESULTS/ANTICIPATED RESULTS: The landscape survey of Apprentice programs revealed multiple models in use. The newly formed Apprentice subgroup is engaging in analysis and actively working to build a standardized repository of competency-aligned, research courses and experiences for apprentices. Results will help make the business case for starting or growing programs. Subgroup members have focused on a shared goal of expanding opportunities for underrepresented minority applicants, with current outreach efforts that are extending awareness of the CRP profession. We anticipate a continuous strengthening of connections between institutions to share a variety of models to implement, develop shared tools (e.g., proficiency tests), and share existing tools to standardize pathways and training for CRP apprenticeships. #_msoanchor_1 DISCUSSION/SIGNIFICANCE: Academic Medical Centers (AMCs) need novel strategies to support clinical research portfolios.Innovative Apprenticeship Models improve efficiency and sustainability of the clinical research professional (CRP) workforce to train the next generation of CRPs in an effective and timely way.

OBJECTIVES/GOALS: Previous literature has suggested that the lack of participation of minorities and women in the science field affects research centers’ quality and competitiveness. This study assessed perceived self-efficacy, obstacles and factors that influenced the development of Hispanic women in a STEMM career. METHODS/STUDY POPULATION: This project followed a mixed-method approach with a convergent design. It included two simultaneous phases: an electronic survey as the quantitative phase and semi-structured interviews as the qualitative phase. Data was collected from an academic-research institution in Puerto Rico. A total of 71 questionnaires were collected applying the Baessler & Schwarzer General Self-efficacy Scale. Five women were interviewed using a semi-structure questionnaire. These women hold a tenure-track faculty position, have held a leadership position, had more than ten publications in peer-reviewed journals, and have received grants for research projects. The data was analyzed by triangulation. RESULTS/ANTICIPATED RESULTS: The General Self-efficacy scale sum scores ranged from 26 to 40 and a mean score of 34.25 (SD=3.79). Items where all answered the higher score stated, ’Thanks to my qualities and resources I can overcome unforeseen situations’ and ’I can solve most problems if I put in the necessary effort.’ Age was the only variable showing statistically significant relationship with general self-efficacy. The most relevant factor facilitating the development in STEMM fields’ careers was the family support. Other contributing factors were the opportunities of employment, positions, professional development, financial status, early experiences during middle and high school, and good mentors, among others. DISCUSSION/SIGNIFICANCE: Results identified some contributing factors and barriers for Hispanic women’s career development in STEMM. Strategies must be implemented as early as middle school for girls showing interest in this field and should include administrative resources to support the career development of women.

OBJECTIVES/SPECIFIC AIMS: In a randomized controlled trial in participants with HIV infection, recombinant human growth hormone (rhGH) reduced visceral adipose tissue (VAT); addition of rosiglitazone to rhGH prevented the accompanying decline in insulin sensitivity (SI). Within this parent RCT, we sought to determine the effect of rosiglitazone and rhGH intervention on alpha-1-acid glycoprotein (AGP), a biomarker of inflammation. We also investigated AGP as an independent risk factor for SI and VAT changes along with any potential effect modification by AGP of the intervention. METHODS/STUDY POPULATION: Participants with HIV-infection (n = 72) with abdominal adiposity and insulin resistance were randomized to rosiglitazone, rhGH, combination, or placebo for 12 weeks (NCT00130286). SI was determined by frequently sampled intravenous glucose tolerance test, and VAT by whole body MRI. AGP concentrations were determined by immunoturbidimetric assay in available serum samples at baseline (time 0), 4, and 12 weeks (n = 41 participants with samples at all 3 time points). A linear mixed model was used to assess the impact of intervention over time on AGP concentrations. General linear models were used to assess baseline AGP concentrations as an independent predictor of SI and VAT changes by treatment group with the model initially including age quartile, gender, race, ethnicity, BMI, HIV RNA <400 copies/mL, antiretroviral regimen, CD4 count, Stavudine use, and zidovudine use with step-by-step removal of least significant predictors. Effect modification was assessed by adding an interaction term between AGP and assigned intervention. RESULTS/ANTICIPATED RESULTS: AGP did not differ among treatment groups at baseline; overall median (Q1, Q3): 0.608 (.526,.727) g/L, P = 0.92. Treatment with rosiglitazone, rhGH, or the combination significantly reduced AGP concentrations from baseline to week 12, compared to placebo (time by treatment interaction, P = 0.0038). Baseline AGP was not a significant predictor or effect modifier of SI change in response to treatment (P ≥ 0.50). Baseline AGP (g/L) was an independent predictor of VAT change (L) (β = 1.91, SE = 0.89, P = 0.038) in addition to a treatment effect (P < 0.001) and age quartile effect (P < 0.001). No other predictors or interactions were significant, including effect modification of AGP (AGP by treatment interaction P = 0.50). DISCUSSION/SIGNIFICANCE OF IMPACT: It is known that immune and metabolic pathways are highly integrated, and biomarkers of inflammation have predictive abilities for cardiovascular and metabolic disease outcomes. This analysis provides data showing that treatment with rosiglitazone or rhGH in the context of HIV reduces AGP concentrations, indicating efficacy in reducing systemic inflammation. Baseline AGP was an independent risk factor for VAT changes as those with lower AGP at baseline showed a greater reduction in VAT in response to treatment. Biomarkers of inflammation may provide prognostic information for individualized patient outcomes to help guide treatment and follow-up.

OBJECTIVES/GOALS: Minority faculty have inequitable access to information, professional development, and research resources. A structured research-mentoring program could help strengthen the research acumen of underrepresented (UR) faculty, provide a community, and support to ensure their success in becoming independent investigators. METHODS/STUDY POPULATION: The Translational Research Institute (TRI) STARs program aims to build a peer support community of UR in biomedical, clinical, behavioral and social sciences to support career development and research success. The program provides a structured peer support group with a 3-month grant training and development program and addresses issues of isolation often felt by UR faculty in academic settings. It encourages the development of innovative research ideas in a safe environment. This peer support group can also help improve confidence and self-efficacy in clinical and translational research development and execution by UR faculty. At the didactic program’s conclusion and seed grant application submission, STARs provides $10,000 as a TRI DEI Equity, Diversity, and Grantsmanship Expertise project. RESULTS/ANTICIPATED RESULTS: Since its launch in 2021, 11 scholars have enrolled in the program;three have fully completed the program, and all three have received subsequent grant funding. Four scholars have completed the didactic program and are in the process of using seed funding to collect initial data and working on initial publications. The remaining scholars are currently in the didactic program. Initial scholar satisfaction with the program is high: 100% reported satisfaction with their participation (Very Satisfied/Satisfied), and 100% agree the program provides adequate support to their research project (Strongly Agree/Agree). Overall, scholars reported an average increase in confidence of 7.9% in grantsmanship skills (Scale 0-10). The return on investment is 3106%, with over $1.9 million in subsequent funding. DISCUSSION/SIGNIFICANCE: Research shows diverse teams working together, capitalizing on innovative ideas, and distinct perspectives outperform homogenous teams. Our preliminary experience demonstrates success for the model. Additional, long-term support will be furthered developed to address additional challenges experienced by UR faculty across their careers.

OBJECTIVES/SPECIFIC AIMS: The objective of our collaboration is to develop a strong transdisciplinary team consisting of microfluidics engineers, cancer biologists, and clinicians, to identify cell surface markers capable of detecting circulating osteosarcoma cells (COC) using microfluidic devices. Our goals are 3-fold: (1) Identify cell surface markers unique to osteosarcoma (OS) for COC isolation, (2) develop a Geometrically Enhanced Mixing (GEM) device to isolate COCs, and (3) Evaluate the efficacy of GEM device to detect COCs in OS patients under treatment. The long-term goal is to utilize this cell detection approach to correlate the presence of COC with metastatic incidence. METHODS/STUDY POPULATION: To identify a marker to capture COCs we are utilizing flow cytometry and microfluidic capture devices. Flow cytometry will be used to evaluate the relative expression of epithelial cell adhesion molecule (EpCAM), CD45, cell surface vimentin (CSV), insulin-like growth factor 2 (IGF2R), interleukin 11 receptor subunit alpha (IL-11Rɑ), ganglioside 2 (GD2), and receptor activator of nuclear factor κ-B (RANK) on a panel of OS cell lines. These cell surface markers were selected based on an extensive review of OS cell surface markers. OS cell capture efficacy will be assessed by passaging a known concentration of OS cells through a GEM microfluidic device coated with antibodies targeting the selected marker, as indicated by flow cytometry. Once captured, COCs on the device will be analyzed and the capture efficiency for the indicated marker will be measured. ANOVA will be used to determine any significant difference in capture efficiency between marker types. Once an optimal marker or panel of markers has been selected we will conduct capture studies using OS cell spiked blood samples followed by clinical samples obtained from OS patients. In clinical samples, COC detection will be validated using the FDA approved triple immunocytochemistry technical definition of a circulating tumor cell (CTC). This will enable COCs to be differentiated from the normal whole blood cell population by selecting for CD45−, EpCAM+, and cytokeratin+ cells. RESULTS/ANTICIPATED RESULTS: Our preliminary studies have shown that on our microfluidic device, EpCAM, a marker commonly used to identify circulating tumor cells in other cancer settings, has a poor capture efficiency (15.9%+7.7%) for HU09 OS cells while the same setup with EpCAM has a capture efficiency of 56.9%+2.7% for BXPc-3 pancreatic cells. We therefore anticipate our flow cytometry studies to show a low expression of EpCAM and CD45 for OS cell lines, while showing a moderate to high expression of CSV, IGF2R, IL-11Rɑ, GD2, and RANK. We expect to show a 60%–80% capture efficiency for markers selected for COC capture. Currently, CSV and GD2 are particularly promising as markers based on previously published studies. DISCUSSION/SIGNIFICANCE OF IMPACT: OS is the most common primary bone tumor and the third leading cause of pediatric cancer deaths. At diagnosis 80% of patients will present with metastasis, however only 20% of these cases are clinically detectable. Innovative strategies to identify patients at risk of metastasis would allow for stratification of intervention therapies. Currently, tumor recurrence and metastasis are primarily dependent on diagnostic-imaging modalities such as computerized tomography or positron emission tomography scans. Unfortunately, these imaging modalities can only detect tumor masses of significant size (106 tumor cells). Liquid biopsies are a novel alternative to current diagnostic imaging systems to monitor metastatic incidence and treatment efficacy. The detection of CTCs through routine blood sampling has the potential to be used clinically for earlier detection, monitoring the treatment of metastatic cancers and surveying the effect of therapeutic interventions on metastasis. To date, the majority of the studies on CTCs have evaluated their presence in carcinomas. Although sarcomas are rare, they generally have a poor prognosis. This study will address one of the unmet medical needs in the field of CTC detection; the identification of cell surface OS makers to improve binding specificity, increase purity, and maintain a high capture efficiency. This phase of our proposal will evaluate the most abundant and conserved markers across a panel of OS cell lines. Once a marker or panel of markers is selected, we will begin to develop a microfluidic device that can be used clinically to detect CTCs in this disease setting.

OBJECTIVES/GOALS: Damage to the sensorimotor cortex areas or/and motor/sensory pathways after a stroke could lead the motor system to a loss of controllability for joints. We investigate the loss of individual joint controllability called a loss of individualization during arm movement, which would provide an insight into abnormal motor coordination. METHODS/STUDY POPULATION: We recruit 12 chronic stroke survivors with Fugl-Meyer score between 26 and 50. A robotic exoskeleton with minimum mechanical resistance is equipped to measure the movements of the shoulder, elbow and wrist joints, respectively. Surface EMGs on muscles related to the joints are recorded using 11 wireless pre-amplified electrodes. Participants are asked to move the shoulder, elbow, or wrist joint individually throughout their range of motion, without moving the other joints voluntarily. RESULTS/ANTICIPATED RESULTS: It would be expected that participants show more difficulty in individualization of the distal joint in comparison with the proximal joint. A reduced joint range of motion would be observed in a descending order of the wrist, elbow and shoulder. These results are in line with the proximal-to-distal gradient of motor deficits after a stroke. Intention of moving the distal joint would induce a greater deviation in the position of the proximal joint than that of the distal joint when moving the proximal joint. A non-negative matrix factorization algorithm would reveal a decreased number of muscle synergies in the groups with a loss of individuation in comparison with the groups with no loss. DISCUSSION/SIGNIFICANCE OF IMPACT: We demonstrate that a stroke leads to a lack of individual joint controllability, with a greater deficits on the distal joint, and that it is related to a decreased number of muscle synergies across the corresponding joints. CONFLICT OF INTEREST DESCRIPTION: N/A.

ABSTRACT IMPACT: This research highlights Adverse Childhood Experiences as a potential risk factor and intervention target contributing to the disproportionate number of individuals with sickle cell disease affected by asthma, a well-established catalyst to the increased morbidity and mortality impacting this high-risk population. OBJECTIVES/GOALS: Adverse Childhood Experiences (ACEs) are strongly associated with asthma. A disproportionate number of individuals with sickle cell disease (SCD) also have asthma. Asthma is strongly associated with increased SCD morbidity and mortality. This study compared the prevalence of asthma among children and adolescents with SCD with and without ACEs. METHODS/STUDY POPULATION: This retrospective cohort study involved 45 children and 30 adolescents with SCD. ACEs were captured using the Center for Youth Wellness Adverse Childhood Experiences Child and Teen Questionnaires, which encompass the original 10 ACEs as well as 7 (child) and 9 (teen) expert-recommended ('expanded') ACEs. ACE exposures were categorized as: Original 0-1 vs. ≥2; Original + Expanded 0-1 vs. ≥2. Asthma prevalence was compared among ≥2 and 0-1 ACE groups using the chi-square (or Fisher’s exact) test. A binary logistic regression was performed to predict the likelihood of asthma while adjusting for characteristics (age, household income and gender) that were statistically different among ACE comparison groups at baseline. RESULTS/ANTICIPATED RESULTS: Among the 45 child participants, 64% had a history of asthma; whereas 50% of teens had a history of asthma. Asthma prevalence was higher among teens with ≥2 vs. 0-1 Original ACEs (89% v. 33%, p=0.014). A history of ≥2 ACEs remained significant (p=0.024) among teens after adjusting for age, household income and gender. There was no significance in asthma prevalence among child ACE comparison groups. DISCUSSION/SIGNIFICANCE OF FINDINGS: Adolescents with ≥2 ACEs had a higher prevalence of asthma compared to subjects with 0-1 ACE. This study, coupled with the cumulative nature of ACEs and the graded-dose response relationship between ACEs and poor health outcomes, highlight the need for larger, longitudinal studies examining the relationship between ACEs, asthma and SCD outcomes.

ABSTRACT IMPACT: Our work provides guidance on whether dietary fat intake influences serum cholesterol levels in response to ketogenic diet therapy in adults with epilepsy. OBJECTIVES/GOALS: The modified Atkins diet (MAD) is used in the management of drug-resistant epilepsy in adults. Some patients on MAD show an increase in serum levels of total cholesterol and low-density lipoprotein (LDL) cholesterol. We explored whether dietary fat composition predicts short-term elevations in serum lipid levels in diet-naive adults who begin MAD. METHODS/STUDY POPULATION: Participants self-reported their diet intake with 3-day food records at baseline, 1 month and 2 months. Food records were analyzed using Nutrition Data System for Research software. Fasting serum levels of total cholesterol (TC), high-density lipoprotein (HDL) cholesterol, and triglycerides were also collected and LDL level calculated at baseline, 1 month, and 2 months. RESULTS/ANTICIPATED RESULTS: 38 patients submitted complete food records at each study visit (baseline, 1 month, and 2 month). Compared to baseline diet intake, there was a significant reduction in daily carbohydrate intake at 1 and 2 months (p<0.001) and a significant increase in daily fat intake at 1 and 2 months (p<0.001). There was also a significant increase in daily saturated fatty acid (SFA) intake at 1 and 2 months (p<0.001), daily mono-unsaturated fatty acid (MUFA) intake at 1 and 2 months (p<0.001), and daily cholesterol intake at 1 month (p<0.05) and 2 months (p<0.001), but no change in daily poly-unsaturated fatty acid (PUFA) intake over time. Compared to baseline, there was a significant increase in serum LDL at 1 month (p<0.001) and 2 months (p<0.01) and an increase in serum TC at 1 month (p<0.01) but not 2 months. DISCUSSION/SIGNIFICANCE OF FINDINGS: Despite a significant increase in total fat, saturated fat and mono-unsaturated fat intake as well as an increase in total cholesterol and LDL levels following MAD initiation, dietary fat composition appears to minimally predict serum lipid values in the short term.