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: We investigate how the gut microbiome protects against arsenic toxicity, showing antibiotic perturbation increases toxicity and causes interindividual susceptibility to a sepsis-like disease state in mice. Here, we aim to understand how baseline microbiomes from various mouse vendors impact these outcomes and characterize the observed disease. Methods/Study Population: We developed a novel mouse model where mice are exposed to an antibiotic (cefoperazone) for 2 days, followed by co-exposure to the antibiotic and 100 ppm arsenate. So far, we have evaluated C57BL/6N mice from MSU’s in-house colony, Taconic Biosciences (TAC), and Jackson Labs (JAX), along with C57BL/6J mice from JAX. To determine if the baseline microbiome drives inter-vivarium differences, we established in-house breeding colonies of TAC- and JAX-origin mice at MSU. This allowed us to assess whether, when housed under identical conditions, these mice still show differences in mortality based on their original microbiomes. To characterize the arsenic-induced sepsis-like disease, we performed blood biochemistry assays to quantify the white blood cell populations, and sepsis biomarkers used in clinical settings. Results/Anticipated Results: We observed differences in survival rates between genetically identical mice from MSU (45%), TAC (30%), and JAX (2.5%) in our model. From this, we characterized the baseline composition of the gut microbiomes of these mice and found they were significantly different from each other. We are still awaiting results from our in-house TAC and JAX experiments but expect them to have similar gut microbiome compositions to those directly purchased from TAC and JAX and respond similarly. In our blood biochemistry analysis, we found sick mice presented with low WBC counts and notable biomarkers indicative of liver, heart, and kidney distress. We also anticipate that 16S sequencing results of cecal contents will further support findings by providing evidence of a bacterial infection in the ceca of sick mice. Discussion/Significance of Impact: Collectively, our work demonstrates that antibiotic perturbation of the gut microbiome induces an inter-individual and inter-vivarium susceptibility to an arsenic-induced sepsis-like disease state. This work highlights the importance of considering antibiotic use in the risk assessment of arsenic to better protect the health of those exposed.

Objectives/Goals: Immune checkpoint inhibitors (IO) have dramatically improved survival outcomes in patients with metastatic melanoma. Still, many patients do not respond these treatments, and others may experience harmful adverse events (irAEs). Thus, there an unmet need for biomarkers for real-time monitoring and management of patients exposed to IO therapies. Methods/Study Population: Serial serum samples were collected from patients with BRAFV600-mutant metastatic melanoma treated with ipilimumab/nivolumab (IO, n = 14) or dabrafenib/trametinib (TT, n = 10). Methylated cell-free DNA (cfDNA) was isolated and sequenced using enzymatic methyl-seq. We develop a robust computational pipeline to identify the top 250 cell-type specific regions of differential methylation (DMRs) across 24 cell-types. Using these differentially methylated regions, a deconvolution tool was developed to determine the abundance of cell type-specific cfDNA in patient serum, and changes in abundance were tracked over treatment time-course to assess response treatment and identify signals of adverse events. Results/Anticipated Results: We demonstrated improved precision in DMR detection evidenced by a higher area under the receiver operator characteristic curve (AUROC) of 0.85 on average. Pathway and functional annotation analysis revealed melanocyte-specific methylation marker regions regulated genes related to melanocyte development and differentiation, including MITF, SOX9/10, and FOXD3. We show these regions are conserved through the transformation to malignant melanoma, indicating melanocyte cfDNA abundance can be used as a marker for tumor burden. We characterize the dynamics of melanocyte-derived cfDNA over the course of treatment in responders and nonresponders to both IO and TT. We observe that changes in concentrations of cfDNA from other cell types correlate with clinically observed irAE-mediated damage to normal tissue. Discussion/Significance of Impact: We demonstrated the utility of decoding the origins of cfDNA fragments obtained from serial liquid biopsy samples. Using cell-specific methylation marks, we identified a signature from the primary melanoma to assess response to treatment, while also obtaining a signal from other tissues throughout the body to monitor immune related adverse events.

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/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/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: 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.

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.

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.

OBJECTIVES/SPECIFIC AIMS: Sleep disturbance is a common problem following military deployment. Insomnia is associated with other adverse psychiatric and medical health outcomes. There are specialized cognitive behavioral therapies that can effectively treat insomnia; however, these tend to emphasize dysfunctional beliefs about sleep rather than nocturnal vigilance. Deployment to a threatening environment can engender nocturnal vigilance, which appears to be a salient feature of sleep disturbance in formerly deployed veterans. The purpose of this analysis is to characterize sleep-interfering thoughts and behaviors observed in an ongoing pilot study of a novel 2-session intervention incorporating various cognitive techniques to improve sleep in veterans. METHODS/STUDY POPULATION: To date, 10 formerly deployed US veterans with disturbed sleep have been recruited from the greater DC area. Participants are assessed at baseline, receive 2 intervention sessions, and are again assessed in 3 months. Sleep-interfering thoughts and behaviors are evaluated via self-report forms including the Fear of Sleep Inventory (FoSI), interviews, and prospective diaries. A portion of both intervention sessions addresses vigilant behaviors and sleep-interfering thoughts by teaching participants 1 of 4 techniques that target nocturnal vigilance: cognitive defusion, body scan, self-guided pleasant imagery, and dream rescripting. RESULTS/ANTICIPATED RESULTS: All of the first 10 participants endorsed sleep-interfering thoughts on the Fear of Sleep Index (FOSI) at a severity level of at least “a few times per month” (rating of ≥1), including several regarding previous trauma (#5) and nightmares (#10 and #16). Other elicited thoughts included thoughts about their environment (n=6), sleep (n=5), social or occupational concerns (n=8), nightmares (n=5), and health (n=4). All of the first 10 participants endorsed vigilant behaviors, including being over-attentive to their environment (n=7), checking behaviors (n=6), and being “on-guard” (n=8). Cognitive technique was selected by the participant in collaboration with the facilitator. Customized recommendations were given as to the timing and duration of practice, but all participants were instructed to practice at least once daily. Three participants (n=3) were fully compliant with their cognitive technique recommendations (choosing a body scan or imagery), 5 were partially compliant, and 2 were not compliant (both chose cognitive defusion). There was a significant reduction in sleep onset latency and wake after sleep onset from baseline to post-treatment (p<0.05). DISCUSSION/SIGNIFICANCE OF IMPACT: The preliminary data suggests that veterans exhibit cognitive and behavioral patterns that involve vigilance and interfere with sleep and demonstrates the need for an intervention targeting the link between nocturnal vigilance and sleep disturbance. More veteran participants and feedback are needed to optimize the efficacy and effectiveness of this sleep training.

OBJECTIVES/GOALS: To determine if a multi-modal, interdisciplinary intervention delivered to a group of prediabetic patients will result in reduced rates of diabetes progression. This project is a retrospective evaluation that will exam the feasibility and possibly efficacy of this intervention. METHODS/STUDY POPULATION: We will evaluate outcomes of 50 participants for the clinic, aged 21-60 inclusive. Patients will have a Body Mass Index >25kg/m2 with a diagnosis of prediabetes. Patients must be non-pregnant, using approved contraception, and agree to not become pregnant for 1 year after enrollment. After enrollment, the initial treatment period is for 1 year and includes a 12 week low calorie diet plan, a 6-month intensive behavioral and lifestyle modification plan followed by a 6 month behavior reinforcement extension. Weight management medications may be used if appropriate for the patient from a clinical perspective during the 6-month intensive behavioral/lifestyle modification. RESULTS/ANTICIPATED RESULTS: It is anticipated that there will be decreased weight with a mean weight loss goal of approximately >10%. Furthermore, it is expect that there will be improvement of other markers of metabolic disease. These include improvement of lipid values (LDL-C, HDL-C, Triglycerides, Total Cholesterol) as well as blood pressure with expected blood pressures of below 130/80 in greater than 50% of participants. Finally, It is expected that 50% or greater participants will have improvement of glycemic control. It is anticipated that greater than 50% of participants will have improvement of glycemic control and achieve normoglycemia. These values will be determined based upon fasting glucose or A1c. DISCUSSION/SIGNIFICANCE OF IMPACT: The significance of this intervention is enormous. By demonstrating feasibility in this trial, we can work toward both assessing efficacy and possibly dissemination of this model program. If these interventions provide durable changes at scale, this could help slow the epidemic of obesity and obesity related comorbid conditions.

OBJECTIVES/SPECIFIC AIMS: Engagement in risky behaviors is not uncommon among adolescents. Two factors associated with risk taking are difficulty regulating emotions and impulsivity. Moreover, youth who exhibit higher scores on impulsivity-like personality traits (ie, negative urgency, positive urgency, sensation seeking, lack of premeditation, and lack of perseverance) are at even heightened risk. An effective intervention decreasing risk-taking behavior among adolescent populations in clinical settings is Dialectical Behavioral Therapy for Adolescents (DBT-A), which teaches skills on emotion regulation, distress tolerance, and mindfulness. However, DBT-A has yet to be tested as an intervention for youth in a nonclinical setting. The current study aimed to fill this gap in the literature. METHODS/STUDY POPULATION: A 9-week DBT-A skills group was implemented in a public high school classroom (7th-8th graders; N=41) and an alternative high school for at risk youth (7th-12th graders; n=21). Of the 41 youth from the public high school classroom participated, with preintervention and postintervention data provided by 30 participants (retention of 73%). RESULTS/ANTICIPATED RESULTS: Results found a significant increase in mindfulness skills and marginally significant increase in emotion regulation skills. Although there was not an overall change in risky behavior among participants, those who were higher on lack of premeditation and positive urgency showed steeper improvements on the skills. The second study at the alternative high school is currently underway, with no current results to report. DISCUSSION/SIGNIFICANCE OF IMPACT: This study will demonstrate that DBT-A skills training is feasible in a school-based setting and shows promising preliminary evidence of decreasing risk of engagement in risky health behaviors among adolescents, particularly among high-risk youth.

OBJECTIVES/SPECIFIC AIMS: Clostridium Difficile Infection (CDI), a prevalent cause of diarrhea, is the most notorious hospital-acquired infection, resulting in an alarming mortality and health care utilization rates. Herein, we investigate the impact of cannabis use, which is gaining significant legalization for recreational use, on the risk of CDI. METHODS/STUDY POPULATION: We selected adult records (age ≥ 18 years) from the Nationwide Inpatient Sample 2014, and identified cannabis users and other clinical conditions using ICD-9-CM codes. With multivariate logistic modeling, we generated propensity scores for cannabis users and matched them to non-users in a 1:1 ratio (104,936:104,936). We then estimated the adjusted relative risk (aRR) for having CDI using conditional Possion regression models with generalized estimating equations [SAS 9.4]. RESULTS/ANTICIPATED RESULTS: Among the matched hospitalizations (n=209,872), cannabis usage was associated with a reduced incidence of CDI (505.8[464.7-550.6] vs. 694.9[645.8-747.70] per 100,000 hospitalizations), resulting in a 27% reduced risk of CDI (aRR:0.73[0.65-0.81]; p-value:<0.0001). Non-dependent and dependent cannabis users respectively had 22% and 78% reduced likelihood of CDI when compared to non-cannabis users (0.78[0.69-0.90] & 0.22[0.12-0.40]). Furthermore, dependent users had less risk of CDI compared to non-dependent users (0.28[0.16-0.51]). Comparatively, abusive use of other substances like alcohol and tobacco was associated with increased risk for CDI (1.30[1.13-1.49] & 1.24[1.10-1.40]) DISCUSSION/SIGNIFICANCE OF IMPACT: Unlike alcohol and tobacco abuse which are associated with elevated risk for CDI, cannabis use, is related to a decreased risk of CDI amongst hospitalized patients. Further prospective and molecular mechanistic studies are required to elucidate how cannabis impacts CDI.

OBJECTIVES/GOALS: There is not much known on how to improve informed consent understanding and there are no effective interventions that have been identified to improve understanding rates of information. This study seeks to assess participants understanding of the informed consent. METHODS/STUDY POPULATION: We studied a non-probability sample of 245 participants, 57% female, with age range from 6 to 84, currently enrolled in clinical trials conducted at an urban city, multi CTSA institution. A self-administered questionnaire approved by IRB was utilized. Redcap database was utilized for data entry. The items in the questionnaire reflected understanding of the informed consent (e.g., purpose for the study, participants’ rights, risks, benefits). Participants completed the survey during their first visit to the research centers or on a follow-up visit. Data were collected from July 2018 to November 2019. Data were analyzed descriptively by summary statistics. RESULTS/ANTICIPATED RESULTS: African Americans were 44%, Non-Hispanic Whites were 36%, Hispanic 6%. Others 13%. 52% married, 12% completed High school, 74.8% completed College, 13% less High school. 91% read the form themselves. 99% knew the purpose of the study; 99% knew they could quit the study at any time. While (113) 47% indicated knowledge of the potential risk, only (12)10.6% could not list any associated risk. 98% stated they had information on who to call with questions regarding the study. (204)86% knew of a potential benefit, only (11)5% could not name some study benefit. 38% were unsure/did not know the total number of visits study required of them. 74% knew the duration of the study. DISCUSSION/SIGNIFICANCE OF IMPACT: Extended discussion and more time on one-one by the study teams in this CTSA tend to increase trust. This approach has been reported to be most effective in improving participant understanding of informed consent process and may result in the positive experience.

OBJECTIVES/GOALS: Clinical Research Professional (CRP) shortages and high turnover rates directly impact the conduct of studies. A strategy for improving CRP retention is using Stay Interviews (SI), which are intentional exchanges between CRPs and leadership to enhance transparency and trust. Importantly, SIs are entirely separate from performance evaluations. METHODS/STUDY POPULATION: Representatives from seven academic medical centers formed a collaborative workgroup (WG) as part of the Association for Clinical and Translational Science (ACTS) Clinical Research Professionals Taskforce (CRPT) Special Interest Group (SIG). The purpose of this workgroup was to develop a Stay Interview (SI) toolkit that will empower leadership to retain high quality staff by engaging employees in open and responsive conversations about motivations to stay in their current roles. Tools previously used by collaborating institutions were evaluated and aligned to establish a sharable guide for SI best practices. Training resources to support leaders in effectively using SI tools were also developed to accompany the toolkit. RESULTS/ANTICIPATED RESULTS: Discussions of current tool use among group members highlighted 2 categorical use cases: continuous integration to promote workforce engagement and retention; and targeted use to address acute workforce challenges. The WG identified the need for a standardized conversation guide as well as leadership tools to support effective use of the guide. From the examples collated, the group crafted a 14 question SI guide with additional probing questions that can be tailored to the work environment. Questions fell into 4 key themes: likes/dislikes, motivations, workplace influence on work life, and professional development barriers and opportunities. Anecdotally, SI use in other industries suggests that wide adoption is likely to reduce CRP workforce turnover and improve job satisfaction. DISCUSSION/SIGNIFICANCE: SIs are designed to build trust and strengthen relationships, fostering positive change by acknowledging issues, understanding motivations, and increasing engagement. Leaders can make immediate actions: clearing obstacles, providing new resources, and increasing recognition. Our next step is implementing a pilot to gather workforce metrics.

OBJECTIVES/GOALS: This poster summarizes the development and implementation of research exploring the relationship between sleep and brain health. METHODS/STUDY POPULATION: Three pilot studies and two secondary data analyses were conducted on 20 older adults with coronary artery disease and 30 older adults without major cardiovascular disease. They were recruited for 10 older adults with multiple chronic conditions. The study included interviews, magnetic resonance imaging, and sleep assessment of participants. Data were also gathered from two secondary sources on multiple chronic conditions, sleep, neuroimaging, cognition, and Alzheimers biomarkers. RESULTS/ANTICIPATED RESULTS: The multidisciplinary team was from nursing, medicine, cardiology, psychology, neuroscience, radiology, and data science to address the separate research aims. The pilot studies and secondary data analyses were successfully implemented. The University of Iowa Institute for Clinical and Translational Science, Center for Advancing Multimorbidity Science, and Iowa Neuroscience Consortium supported the collaboration. The teams have found that sleep and circadian rhythms characteristics may be associated with not only coronary artery disease, but also multiple chronic conditions. Having sleep disorders and reduced circadian amplitude can be associated with white matter microstructure and functional connectivity. These collaborations provided multiple funding and publication opportunities. DISCUSSION/SIGNIFICANCE: Interdisciplinary team research is important to enhance translational science. Although challenges were identified, using multiple methods and dataset sources with multidisciplinary team members enabled opportunities to explore multifaced topics related to sleep and brain aging.

OBJECTIVES/GOALS: In 2014, Johns Hopkins Pediatric ICU (PICU) implemented the PICU Up! early mobilization program. Subsequent studies have shown that these protocols increase mobility of PICU patients. Process improvement requires accurate documentation. Our aim is to evaluate differences in nurse documented and actual reported mobility of PICU patients. METHODS/STUDY POPULATION: A quality improvement project evaluating the impact of a simulation-based early mobility training program is being conducted, with initial analysis of pre-intervention data. Inclusion criteria includes children age 1 day to 17 years old admitted to the PICU for ≥ 3 days during a day shift and exclusion criteria includes specific mobility contraindications. Data on the number of daily mobilizations, highest level of mobility achieved during each mobilization, and occurrence of safety events is captured via direct query of the bedside nurse at the end of a 12-hour shift by a research team member using a standard data collection tool. We also recorded documented events in the electronic medical record (EMR). Paired t-tests were performed for continuous data and McNemar’s test for categorical data. RESULTS/ANTICIPATED RESULTS: In total, 50 patients were enrolled between February and April 2023. The nurses reported that patients participated in a median of 5 mobilizations (Interquartile range [IQR] 4-6) in a 12-hour shift, whereas nurses documented in the EMR that patients participated in a median of 1 mobilization (IQR 0-3; P <0.001). On direct query, the nurses stated that a total of 8 individual safety events occurred during mobility, representing a 3% (8/259) safety event rate. In the EMR, the nurses documented 1 individual safety event during mobility, representing a 1% (1/84) safety event rate (P = 0.008). Nurses reported that they mobilized 50% (25/50) of the patients out of bed; however, they documented that they mobilized only 32% (16/50) of the patients out of bed (P = 0.007). DISCUSSION/SIGNIFICANCE: Compared to EMR documentation, nurses report more mobilization of critically ill children during the day, including more out of bed mobilization and safety events. Future nurse education should focus on mobility documentation to ensure that it reflects mobility at the bedside to facilitate process improvement and optimize care for PICU patients.

OBJECTIVES/SPECIFIC AIMS: Aim 1: To determine the true incidence of trauma-related acute respiratory distress syndrome (ARDS) in India. We propose to perform a prospective observational study to determine the incidence of ARDS in India. Aim 2: To perform a preliminary assessment of risk factors for ARDS in the Indian trauma population. We will leverage these findings against the global ARDS data to provide a foundation for further interventional studies. Aim 3: To evaluate the current management strategies and patient outcomes from ARDS in trauma subjects admitted to the Jai Prakash Narayan Apex Trauma Center (JPNATC). These findings will identify areas in need of practice-based performance improvement in ARDS therapies in India. METHODS/STUDY POPULATION: This application proposes an observational study of trauma patients with ARDS, a population that continues to have substantial in-hospital mortality. The approximate number of ICU-admitted trauma cases for the study period is 1700. Specific data elements to be collected include patient demographics, comorbidities, mechanism of injury, Injury Severity Score, risk factors for ARDS, sequential organ failure and assessment scores, vital signs, laboratory values, and evidence-based treatments received, including mechanical ventilation and adjunctive therapies. Outcome data will include discharge location, ICU and hospital length of stay and all-cause mortality. Selection of Subjects: We will include all patients admitted to the JPNATC Trauma and Neurosurgical ICUs intubated and mechanically ventilated and meeting the definition of Berlin definition of ARDS8. We will collect data for a total of 12 months. RESULTS/ANTICIPATED RESULTS: Due to gaps in reporting, the incidence, mortality, and practice-based management algorithms applied in trauma patients suffering from ARDS in India is unknown. We hypothesize that the overall incidence of trauma-related ARDS is higher, and the fraction of patients managed with evidence-based therapies is lower than global reported averages. DISCUSSION/SIGNIFICANCE OF IMPACT: Although the true incidence of ARDS in trauma subjects in India is currently unknown, we suspect that it is much higher than reported. Such data are important in identification of resource allocation including ICU bed and mechanical ventilator availability, particularly in a resource-limited environment. This proposal will aid in the development of research infrastructure at JPNATC, contribute to capacity building, and the establishment of a Clinical Research unit at the Apex Institute. Finally, a provision to develop a consortium and trauma quality improvement program among the existing trauma centers in New Delhi to disseminate important research findings and guidance to the rest of India is a future benefit of the study.