OBJECTIVES/GOALS: Researchers include community health workers and promotoras (CHW/Ps) on research teams to increase community engagement; however, no formal training on research best practices exists for this group. Study objectives were to examine perceived relevance of a new culturally and linguistically appropriate CHW/P training and optimal delivery modes. METHODS/STUDY POPULATION: We conducted six focus groups (FGs), three each in English and Spanish, at three study sites, University of Florida, University of Michigan, and University of California Davis from February to August 2021. The CHWs/Ps were purposively selected to include diverse age, race/ethnicity, educational level, and work experience. Separate FGs were conducted for CHWs/Ps in English and Spanish as appropriate. All FGs were audio recorded, translated to English from Spanish, transcribed and analyzed using RADaR (Rigorous and Accelerated Data Reduction) technique. RESULTS/ANTICIPATED RESULTS: Forty CHWs/Ps (95% women, mean age 45) participated, with the majority (58%) identifying as Hispanic/Latino. Of the sample, most identified as White (50%) or Black (25%). The proposed training was mentioned as relevant and would help them to be confident, comfortable, knowledgeable and effective in the community. Online training, though advantageous due to its flexibility also reportedly had barriers such as internet access, computer availability and technological know-how of CHWs/Ps. A hybrid training approach, online plus peer-led, was recommended due to the importance of personal guidance by an experienced CHW/P’ especially for a newly recruited CHW/P. DISCUSSION/SIGNIFICANCE: Findings indicated that a culturally and linguistically appropriate CHW/P training that is flexible and easily accessible in its mode of delivery is relevant and useful. In-person guidance to a new CHW/P was reported as an important training component. Poster will include the detailed quotes on relevance, usefulness, and mode of delivery of training.

OBJECTIVES/SPECIFIC AIMS: This study aims to determine if a bronchiolitis specific microbiome exists and how it evolves through disease course. Objective 1. Determine the microbiome profile of the airway in virus induced bronchiolitis-associated respiratory failure. Objective 2. Identify changes in the airway microbiome profile through the course of virus induced bronchiolitis associated respiratory failure, and the relationship between microbiome composition and clinical respiratory status. Objective 3. Determine the impact of rhinovirus infection on the lung and stool microbiome in a murine asthma model. METHODS/STUDY POPULATION: Objectives 1 &2: We are conducting a single-center prospective case-control study of patients admitted to the Komansky - Weill Cornell Pediatric Critical Care Unit. Infants less than two years of age with a diagnosis of bronchiolitis requiring intubation and mechanical ventilation are enrolled as subjects. Infants less than two years of age intubated and requiring mechanical ventilation without primary lung pathology are enrolled as controls. To evaluate our primary objective, tracheal aspirates will be collected from both subjects and controls on the day of intubation. We will perform 16s RNA sequencing on the tracheal aspirate samples and compare the resulting microbiomes. To evaluate secondary objective, we will collect tracheal aspirates of our study population on a daily basis and map the microbiome in parallel with objective measures of respiratory status including oxygen index and successful extubation. Both subjects and controls are being enrolled as a convenience sample. Objective 3: Mice, heterozygous for the sptlc2 gene (Spltc2 +/−) demonstrate reduced de-novo sphingolipids and increased airway hyperresponsiveness with methacholine challenge. Airway hyper-responsiveness is a cardinal feature of asthma. This airway hyperresponsiveness is exacerbated in the setting of rhinovirus (Figure 1). Using 16s sequencing, we will examine the lung microbiome of Sptlc2 +/− ad Sptlc2 +/+ at 1- and 7-days following rhinovirus infection. RESULTS/ANTICIPATED RESULTS: This clinical study is currently IRB approved and enrollment is ongoing. We have enrolled 12 subjects and 5 controls. Sample analysis will begin following the 2018-2019 respiratory season, with an anticipated cohort of 20 subjects and 20 controls. With regards to the murine studies, we have demonstrated that the lung microbiome of Sptlc2 +/− and Sptlc2 +/+ mice is similar at baseline (Figure 2) and remains similar following 1-day infection with rhinovirus. We do however, see a distinct change in the microbiome profile of the stool of Sptlc2 +/− mice following rhinovirus infection (Figure 3). Lung analysis at day 7 post infection is pending. DISCUSSION/SIGNIFICANCE OF IMPACT: These studies will lay the groundwork for detailing the functional role of the airway microbiome in bronchiolitis, with the objective of developing new modalities for disease treatment and prevention. In addition our murine studies allow us to view the microbiome in the context of sphingolipid deficiency, providing a potential mechanistic link to rhinovirus and ORMDL3 associated asthma.

OBJECTIVES/SPECIFIC AIMS: CFTR is the mutant protein that causes cystic fibrosis (CF), a fatal respiratory diseases affecting 1 in 3500 children. CFTR modulators are small molecules that directly address mutant CFTR function. Improving correction of the F508del CFTR mutation (affecting 90% of CF patients) is one of the most pressing unmet needs in CF. Currently available F508del therapeutics only marginally improve CF, In vitro, we have identified a miRNA that impairs utility of CFTR directed therapies. miR-145 is upregulated by TGF-β (a genetic modifier of CF lung disease) with a direct binding site on the 3’-untranslated region of CFTR mRNA. Binding of miR-145 to CFTR destabilizes mRNA transcript and impedes protein translation. Overexpression of miR-145 abolishes benefit of F508del CFTR correction. Antagonists to miR-145 block TGF-β suppression of CFTR function and augment response to CFTR correction. This project evaluate in vivo impact of TGF-beta and miRNA manipulation on CFTR functional readouts including nasal potential difference (NPD) and short circuit current (Isc) across tracheal explants in addition to standard biochemical measures. METHODS/STUDY POPULATION: Wild-type Sprague-Dawley rats were inoculated with an adenoviral vector containing bioactive TGF-beta or sham at 1×109 pfu/animal placed in the left nares. Seven days post-inoculation, functional, and biochemical measures were conducted. NPD was measured with a microelectrode placed in the left nare and grounded the tail. The nare was sequentially perfused with standard Ringer’s solution, amiloride (to block the ENaC sodium channel), low chloride Ringer’s (to stimulate chloride efflux), forskolin (to open the CFTR channel) and CFTRinh-172 (to block the CFTR channel. Tracheal explants were harvested, microdissected, and placed on modified Ussing chambers. RESULTS/ANTICIPATED RESULTS: We have inoculated WT rats with bioactive TGF-β Versus sham delivered by intranasal inoculation of an adenoviral vector. Functional readout of CFTR function is by Isc across tracheal epithelia and NPD. Lung homogenates are analyzed for TGF-β signaling, miRNA expression, and CFTR transcripts. Both tracheal explants and NPD indicate TGF-β stimulation diminishes CFTR function in vivo. In tracheal explants, TGF-β exposure diminishes CFTR response to forskolin-stimulation by 75%. Loss of current after CFTR inhibition (CFTRinh-172) is halved. By nasal PD, TGF-β inoculation similarly halves the bioelectric response to low chloride and forskolin stimulation. Evaluation by qPCR reveals a strong increase in TGF-β signaling demarcated by PAI-1, prompting a reduction in CFTR mRNA. miR-145 is expressed highly in rat pulmonary tissue, but no change in overall miR-145 levels was detected between TGF-β and sham exposed rats. This finding reflects what we have observed in human lungs, with a localized increased miR-145 expression in CF epithelia, but similarly high levels of miR-145 in both CF and non-CF whole lung homogenates. Although expressed at lower levels than miR-145, we did find increased expression in TGF-β relevant miR-101, miR-494, and miR-144 that have a predicted binding site on rat 3’-UTR in TGF-β exposed Versus sham lungs. DISCUSSION/SIGNIFICANCE OF IMPACT: Our data indicate the relevance of TGF-β stimulation to suppress CFTR synthesis and function in vivo. Future work will evaluate whether these additional miRNA with CFTR binding sites may mediate TGF-β suppression of CFTR in the rat model, and the utility of miRNA manipulation to augment F508del CFTR correction.

OBJECTIVES/GOALS: Adoption of the Observational Medical Outcomes Partnership (OMOP) common data model promises to transform large-scale observational health research. However, there are diverse challenges for operationalizing OMOP in terms of interoperability and technical skills among coordinating centers throughout the US. METHODS/STUDY POPULATION: A team from the Critical Path Institute (C-Path) collaborated with the informatics team members at Johns Hopkins to provide technical support to participating sites as part of the Extract, Transform, and Load (ETL) process linking existing concepts to OMOP concepts. Health systems met regularly via teleconference to review challenges and progress in ETL process. Sites were responsible for performing the local ETL process with assistance and securely provisioning de-identified data as part of the CURE ID program. RESULTS/ANTICIPATED RESULTS: More than twenty health systems participated in the CURE ID effort.Laboratory measures, basic demographics, disease diagnoses and problem list were more easily mapped to OMOP concepts by CURE ID partner institutions. Outcomes, social determinants of health, medical devices, and specific treatments were less easily characterized as part of the project. Concepts within the medical record presented very different technical challenges in terms of representation. There is a lack of standardization in OMOP implementation even among centers using the same electronic medical health record. Readiness to adopt OMOP varied across the institutions who participated. Health systems achieved variable level of coverage using OMOP medical concepts as part of the initiative. DISCUSSION/SIGNIFICANCE: Adoption of OMOP involves local stakeholder knowledge and implementation. Variable complexity of health concepts contributed to variable coverage. Documentation and support require extensive time and effort. Open-source software can be technically challenging. Interoperability of secure data systems presents unique problems.

OBJECTIVES/GOALS: High blood-pressure (BP) is a common adverse effect of erythropoietin (EPO) therapy among patients with chronic kidney disease on hemodialysis, and even among otherwise healthy individuals who receive EPO. In human genetics, EPO is associated with not only red blood cell traits, but hypertension (HTN) as well. Currently, there is no vascular gene expression data available in the setting of EPO-induced HTN that may explain precise role of key cellular players in its hypertensive etiology. Our aim is to characterize vascular transcriptome to identify key cellular players in EPO-induced HTN. METHODS/STUDY POPULATION: 10-12 week C57BL/6 male and female mice were randomly divided into two groups 1. Vehicle (0.9% saline-VEH), 2. EPO, (N = 4). VEH and EPO were intraperitoneal administered (EPO 75U/30g, 3 times/week) for 20 days. Blood-pressure was measured non-invasively via tail-cuff plethysmography. We characterized in-vivo transcriptome response of mouse descending aorta to EPO-HTN and vehicle control group by high-throughput RNA-sequencing. RESULTS/ANTICIPATED RESULTS: Systolic blood pressure (SBP) was significantly higher in EPO treatment, compared to vehicle (males and females combined SBP VEH 116.29 ± 6.21, EPO 129.57 ± 4.59, mean ± s.d., adjusted P = 0.0012). Comparison of in-vivo transcriptional differences between vehicle and EPO-treated reveal statistically significant changes in cellular pathways consistent with hypertension such as upregulation of RAS signaling pathway and oxidative stress. In-vitro mouse aortic smooth muscle cells, EPO markedly increased phosphorylated-ERK activity, suggesting increased RAS activity. DISCUSSION/SIGNIFICANCE OF IMPACT: This study highlights the importance of previously unknown vascular key players and advances our understanding of the transcriptional events associated with EPO-induced hypertension.

ABSTRACT IMPACT: Predicting therapeutic responses in GBM. OBJECTIVES/GOALS: The goal of this team approach is to integrate mathematical models of glioblastoma (GBM) infiltrating myeloid cells that contribute to the immunosuppressive phenotype in glioma with experimental data to predict therapeutic responses to combined chemokine receptor and immune checkpoint blockade. METHODS/STUDY POPULATION: Orthotopic murine KR158-luc gliomas were established in fluorescent reporter CCR2WT/RFP CX3CR1WT/GFP mice. Subsequently, an anti-CD31 injection was administered to label the vasculature. Fluorescent imaging and quantification of anti-CD3 stained sections were performed on a range of tumor sizes to acquire vasculature, tumor, T cell, and myeloid cell densities. In parallel, a system of ordinary differential equations was formulated based on biological assumptions to evaluate the change over time of tumor cells, T cells, and infiltrating myeloid cells. The model was then refined and validated by experimental results. RESULTS/ANTICIPATED RESULTS: Fluorescent imaging and quantification revealed a correlation between tumor size and abundance of (CX3CR1+, CCR2-) and (CX3CR1+, CCR2+) myeloid cell populations in the tumor microenvironment. The density of these cell populations and vasculature remained constant as the tumors increased in size. Computer simulations of the mathematical model will predict tumor, myeloid, and T cell dynamics. These simulations will be particularly useful to uncover information regarding myeloid cell dynamics, such as cell entry time into the tumor microenvironment. Parameter sensitivity analysis of the model will inform us of the biological processes driving these tumor-immune cell dynamics. DISCUSSION/SIGNIFICANCE OF FINDINGS: GBM is a challenge as current intervention are ineffective. This study improves the understanding of glioma infiltrating myeloid cells and their impact on tumor progression. The data will serve as a basis for quantitatively predicting therapeutic responses of a novel combination treatment.

OBJECTIVES/SPECIFIC AIMS: To share lessons learned from implementing a health survey to a global sample of mTWs. METHODS/STUDY POPULATION: mTWs were paid $0.50 for taking a 15 minute survey to ascertain attitudes and intentions toward participating in genetic research. Two phases included: pilot survey targeting 7 global regions and a large-scale implementation in English in United States, India, and other countries and in Spanish in Spanish speaking countries. Administrative and descriptive information were collected and analyzed by region/country including: completions by location, demographics, time to complete, and survey satisfaction. RESULTS/ANTICIPATED RESULTS: There are 4 key lessons: (1) MTurk is fast. The US sample (n=505) accrual took <2 days and the Indian sample (n=505) took 11 days, while the response from other countries (n=118) generally exceeded 30 days. (2) Using Amazon country specification was the best way to ensure responses from specific countries and regions. (3) Demographic differences exist in mTWs between countries. For example, US mTWs were significantly more likely female (60.1%) compared with India (30.2%) and other countries (34.2%). (4) mTWs found the survey understandable/acceptable. mTWs reported high understandability and acceptability of the survey, which did not vary significantly across countries or by language. DISCUSSION/SIGNIFICANCE OF IMPACT: mTurk provides an efficient platform for survey research from diverse US and Indian samples. In other countries and in Spanish, the mTurk mechanism yielded a smaller sample more slowly but was still effective.

OBJECTIVES/GOALS: The Indiana CTSI is a partnership with Indiana University, Purdue University, University of Notre Dame, and Regenstrief Inst. IU’s Comprehensive Cancer Center is central to cancer research and education. A partnership between these critical entities ensures certain efficiencies. We provide a potential framework for community outreach efforts. METHODS/STUDY POPULATION: The Indiana CTSI’s partner institutions have long prioritized community outreach and engagement across the state. However, in environments with limited funding resources, efficiencies are critical to the sustainability of programs and efforts. All IN for Health, an initiative of the Indiana CTSI, has partnered with IU Simon Comprehensive Cancer Center in community outreach by evaluating current practices, aligning staffing, evaluating events, prioritizing outreach efforts, and strategic outcomes. A tool for evaluation was developed and the prioritization matrix along with a database of events now guide outreach efforts. The All IN for Health board continues to be highly engaged in providing feedback and developing strategies. RESULTS/ANTICIPATED RESULTS: This partnership has increased outreach to state-wide events, including urban and rural communities, as well as events contributing to the health of historically marginalized groups in the state. The challenge was our ability to be present at all community events that are critical to the success of all our partners, but most importantly the communities we serve. Opportunities to partner across non-academic and community health partners were evaluated with an assessment of All IN for Health efforts. The resulting approaches are used as an example or a potential framework from which to organize similar partnerships with the goal of advancing research and health equity. Through this partnership, we have extended outreach and added efficiencies, demonstrating creative implementation and strategies. DISCUSSION/SIGNIFICANCE: Unfortunately, limited funding prevents CTSAs and Cancer Centers from engaging everywhere they are needed. Translation research constantly encourages team science and collaboration. Our efforts are a reminder that the same approach applies to operations and synergizing the assets present within our community health and institutional partners.

OBJECTIVES/GOALS: Primary care practices struggle to identify which combination of care structures and processes need to be implemented to improve practice performance and subsequently, patient outcomes. The goal of this study is to develop and validate a tool to assess care structures and processes that are associated with better quality and patient outcomes. METHODS/STUDY POPULATION: Data from a scoping review, Delphi study, and qualitative interviews with high-performing primary care practices contributed to the development and content validation of the Tool for Advancing Practice Performance (TAPP). From these sources we identified 314 items representing care structures (e.g., care team makeup, use of electronic health records) and processes (e.g., care coordination, panel management). We developed criteria for deleting and rescuing items and received input from our expert panel to refine the pool of items. We eliminated items that were redundant and lacked clarity/specificity. The tool was further modified based on feedback from cognitive interviewing and pilot testing with practice managers, quality improvement leaders, and physicians from primary care practices. RESULTS/ANTICIPATED RESULTS: The pool of 314 items was winnowed to 188 after applying criteria for deleting and rescuing items. During the expert review, 70 items were eliminated and 8 new items were added, resulting in a working tool of 126 items. We conducted eight cognitive interviews with the 126-item tool and received feedback on the content, item structure, and language, which led to the elimination of 13 items that were poorly or incorrectly understood by respondents. We also modified the language of 23 items for clarity. After cognitive interviewing, the resulting tool comprised 113 items. Fifteen practices piloted the tool and no additional items were eliminated. We modified the instructions for completing the tool and resolved technical issues related to online administration. DISCUSSION/SIGNIFICANCE: TAPP is a novel tool for assessing care structures and processes that are associated with better quality and patient outcomes in primary care settings. The tool can be used by researchers and primary care clinicians to identify areas for improvement in practice performance and patient outcomes related to chronic disease prevention and management.

OBJECTIVES/GOALS: The Hispanic-In-Research Capability Endowment Phase (HiREC) Phase I Award supports research academic formation of researchers in Puerto Rico. Since 2011, HiREC provided critical infrastructure resources to develop the Clinical and Translational Research (CTR) workforce. We explored the awardees research career advancements and outcomes. METHODS/STUDY POPULATION: The target population of Phase I Award was young faculty and early-career investigators admitted to the Postdoctoral Master of Science in CTR Program. Eleven awardees were supported from 2011to 2020. A survey was administered via REDCap platform and all awardees were invited by email to participate. Data was collected from 03/15/21 to 05/15/21, including weekly follow-up by email and text messages to increase response rate and 100% responded. Descriptive analyses were performed in SPSS VS 27. The 63.6% was female and 54.6% MD. Most were Hispanic faculty and investigators (36.4%) or were at the clinical practice (36.4%). The 45.5% were affiliated to an academic institution or healthcare entrepreneur RESULTS/ANTICIPATED RESULTS: Awardees top career goals were: combine research & teaching-faculty (36.4%), clinical practice (36.4%) and Principal Investigator (PI) in a research institution (36.4%). The 81.8% of awardees achieved those goals and 18.2% are working-on it. A total of 57 grants were submitted (M=10, SD=10.9) and 42.1% funded. Most awardees submitted grants as PI (45.6%) or Co-I (42.1%). Additional outcomes were: 49 peer-reviewed publications, 67 posters and 21 oral presentations. Awardees expressed that this award: provided resources and necessary tools to support the research career pathway; served to develop essential research capabilities, knowledge and skills; and was a unique opportunity to protected-time for research. DISCUSSION/SIGNIFICANCE: HiREC provided resources, opportunities, and tools to remove barriers and facilitate career pathways of Hispanic investigators. Awardees achieved their career goals, their grant success rate was over 40%; received R-grants awards and published over 45 articles. This award model developed promising Hispanic researchers.

OBJECTIVES/GOALS: Obesity is associated with gut dysbiosis, inflammation, and increased intestinal permeability. Probiotic consumption may reverse these outcomes. The goal of this study is to evaluate the evidence linking probiotic consumption to changes in intestinal permeability in subjects with overweight or obesity. METHODS/STUDY POPULATION: Articles were searched in Pubmed, Web of Science, and CAB Direct through February 2022 using search terms: intestinal permeability, overweight or obesity, and probiotic supplementation. 694 articles were exported, and 289 duplicates were identified. Titles and abstract were screened in the 405 remaining references by two investigators to determine eligibility. Eligible studies had data extracted on study participant characteristics, probiotic strain used, probiotic dosage, length of intervention, and intestinal barrier outcomes. Results were summarized in tabular form based on intestinal permeability response to probiotics. Quality of the studies was assessed based on Cochrane–Risk of Bias’ tool (RoB2). RESULTS/ANTICIPATED RESULTS: Thirteen eligible studies were identified. Probiotic genera included Akkermansia, Bifidobacterium, Lactobacillus, Streptococcus, Lactococcus, and Bacillus. Single strain probiotics were used in 3 studies, while the other 10 used multi-strain formulas. Dosage and length of probiotic supplementation ranged from 2.4 x 10^7 to 5 x 10^10 CFU/person/day and 3 to 26 weeks, respectively. The most widely used gut permeability outcomes were serum lipopolysaccharide (LPS) (n=10) and mixed sugar solution consumption with urine analysis (n=6). Five of the 10 studies reported decreases in serum LPS following probiotic consumption, while the other 5 showed no effect. Urinary lactulose/mannitol ratio decreased in 1 of the 4 studies, and urinary lactulose percent decreased in 2 studies. DISCUSSION/SIGNIFICANCE: Probiotic supplementation may be remediating an obesity-induced increase in intestinal permeability as evidenced from the effect on serum LPS and mixed sugar solution assays. However, additional studies are needed to further clarify which strain of probiotic bacteria is most effective and the optimal intervention length in subjects with obesity.

OBJECTIVES/GOALS: The role of the vaginal microbiome (VM) in HIV disease progression is poorly understood. We examined VMs of HIV+ Elite Controllers (ECs) and HIV+ Long-Term Non-Progressors (LTNPs) compared to controls: HIV-positive antiretroviral (ARV) treated (HIV+ATs) and HIV-negative women in the Women’s Interagency HIV Study (DC/Chicago/Atlanta sites). METHODS/STUDY POPULATION: VMs were surveyed via both V3/V4 region of 16S rRNA gene amplicon sequencing and metagenomics sequencing in 67 women across 4 study groups: 1) LTNPs (CD4 >500 cells/mL for 5+ years without ARVs) (n = 7) and 2) ECs (HIV RNA <80 copies/mL for 2+ years without ARVs) (n = 8), matched with 3) HIV+ ATs (on ARVs for ≥1 year with CD4 increase ≥100 cells/mm3) (n = 34), and 4) HIV- women (n = 18). Metagenomes were characterized from specimens collected at two time points: 1) vaginal swabs collected 2016-2017 (n = 62) and 2) cervicovaginal lavage collected 2002-2016 (n = 35; DC/Chicago only). We used VIRGO (human vaginal non-redundant gene catalog), a newly developed referencing framework to comprehensively catalog VM gene content, taxonomy and functions. RESULTS/ANTICIPATED RESULTS: Women were 89% African American with a mean age of 46 years (SD 8.8). The most prevalent species were Gardnerella vaginalis (predominant in 34%), Lactobacillus iners (predominant in 21%), and L. crispatus (predominant in 14%). 90% of LTNP and 45% of EC samples were Lactobacillus-dominant vs. 28% of HIV- and 30% of HIV+ATs. L. crispatus and L. iners in ECs/LTNPs had significantly different gene content and greater gene richness vs. controls. G. vaginalis-predominant communities were found in 66% of HIV- and 68% of HIV+ATs, compared to 46% of EC and 0% of LTNP. The G. vaginalis strains present in EC/LTNP also showed significantly lower gene richness and different gene content vs. controls. DISCUSSION/SIGNIFICANCE OF IMPACT: These results suggest unique VM communities among EC/LTNP, and led us to hypothesize that differential regulation of vaginal immunity drives the observed differences. The similarity between VMs of HIV- and HIV+ATs warrants further study. Larger longitudinal VM studies are needed to assess associated functional pathways and understand the etiology of VM association with HIV progression. CONFLICT OF INTEREST DESCRIPTION: The authors have no conflicts of interest to disclose.

OBJECTIVES/GOALS: Vaccination for COVID-19 is a primary public health strategy to control the pandemic. In this study, we examined how various sociodemographic variables influence rural residents intentions to receive the COVID-19 vaccine. We also examined the role of distrust in healthcare organizations in these intentions. METHODS/STUDY POPULATION: sing the electronic medical records of an academic healthcare institution in central Pennsylvania, we obtained names and addresses of patients who had been an inpatient or outpatient within the prior three years, were 18 years or older, and who resided in a community defined as rural by the Commonwealth of Pennsylvania. The survey included three statements about the intent to receive the COVID-19 vaccine, an open-ended question about concerns regarding the vaccine, and validated scales for general trust and for distrust in healthcare organizations. All study variables were summarized to determine their distributions, and then bivariate binomial logistic regression analyses were conducted. Responses to the open-ended question were coded and used as variables in the bivariate analysis. RESULTS/ANTICIPATED RESULTS: Respondents reporting conservative political views were more likely (compared to those liberal political views) to never want to be vaccinated for COVID-19. Those who expressed distrust in healthcare organizations were less likely to want to be vaccinated soon. Conversely those who were more trusting said they were more likely to want to be vaccinated soon. Respondents dominant concerns about the COVID-19 vaccine were that it was new and that the process for its development was rushed. Respondents who believed that COVID-19 was a hoax were unlikely to ever want to be vaccinated, while those who distrusted the process in some way (new vaccine or rushed vaccine) were more likely to want to wait to be vaccinated. DISCUSSION/SIGNIFICANCE: These findings confirm the impact of political orientation on COVID-19 vaccination intention and suggest that distrust in healthcare organizations may prevent people from being vaccinated. These data provide evidence that people delaying vaccination hold different beliefs than those who will never vaccinate.

OBJECTIVES/SPECIFIC AIMS: Scavenger receptor (SR) surface proteins are highly conserved motifs and are implicated in the uptake of nanotherapies. Gold nanoparticles functionalized with DNAzymes (DzNP) represent a promising novel nanotherapy for lung diseases such as asthma, particularly because they can be delivered directly to the lung. Our lab has been studying the therapeutic potential of a DzNP targeting GATA-3, a master transcription factor regulating Th2 inflammation. Although nanoparticle uptake through scavenger receptors has been described in macrophages in other models, the role of SRs in DzNP uptake in the lung is poorly understood. We hypothesize that scavenger receptors mediate DzNP uptake in alveolar macrophages. To begin examining this hypothesis, we examined whether DzNP exposure and uptake regulates gene expression of MARCO and MSR1, two class A scavenger receptors. METHODS/STUDY POPULATION: Using a silver stain, we measured dose dependent DzNP uptake in murine alveolar macrophages (MH-S). Using qRT-PCR, we measured gene expression of scavenger receptors MSR1 and MARCO in murine alveolar macrophages (MH-S) and after 24 hour exposure to 2251 DzNP, a DzNP targeting GATA-3, and dextran sulfate sodium (DSS), a known SR-A blocker. RESULTS/ANTICIPATED RESULTS: 2251 DzNP uptake in alveolar macrophages is dose dependent. MARCO gene expression levels significantly increase in murine alveolar macrophages when cultured with increasing concentrations of 2251 DzNP (10 pM-2 nM) or DSS 25-200 ug/ml) for 24 hours. However, MSR1 gene expression levels have minimal change when exposed to low concentrations of 2251 DzNP and DSS. At higher concentrations of 2251 DzNP and DSS, MSR1 expression levels are decreased. DISCUSSION/SIGNIFICANCE OF IMPACT: Alveolar macrophages exhibit a dose dependent increase in MARCO gene expression levels with increasing concentrations of 2251 DzNP and DSS, but MSR1 gene expression is not affected in a similar fashion. 2251 DzNP-induced increases in MARCO gene expression suggests that 2251 DzNP may facilitate its own uptake through MARCO. 2251 DzNP exposure negatively regulates MSR1 expression at higher doses and suggests that 2251 DzNP may inhibit its own uptake thought MSR1.

ABSTRACT IMPACT: This study examines gray matter volume differences resulting from the bilingual experience in children and adults allowing us to better understand the brains of over half of the world’s population that speaks more than one language. OBJECTIVES/GOALS: Literature is mixed regarding a bilingual advantage in executive control (EC). While it has been shown that young adult bilinguals have greater gray matter volume (GMV) than monolinguals in EC regions, there is behavioral evidence that suggests such difference would be more pronounced in children. METHODS/STUDY POPULATION: Using SPM12 to test this hypothesis, we used a whole-brain t-test to compare GMV in 35 English-speaking monolingual and 20 Spanish-English early (learned both languages before 6 years old) bilingual children. Next, we submitted both groups of children to an ANOVA with 42 English speaking monolingual and 26 Spanish-English bilingual adults to test for an interaction of Language Experience by Age Group at the level of the whole brain. RESULTS/ANTICIPATED RESULTS: e between-group comparison of bilingual and monolingual children, revealed more GMV in bilingual compared to monolingual children in regions associated with EC (right middle and inferior frontal gyri, superior parietal lobule, and precuneus). Our second analysis, an ANOVA comparing bilingual and monolingual children and adults, revealed an interaction in which bilingual>monolingual GMV in children was greater than any bilingual>monolingual GMV (or bilingual=monolingual GMV) in the adult groups in the right superior parietal lobule (BA1). No regions indicated that bilingual>monolingual GMV was more pronounced in adults. DISCUSSION/SIGNIFICANCE OF FINDINGS: These results provide further evidence for GMV differences in early bilinguals in regions associated with EC and indicate that more GMV differences exist between bilingual and monolingual children than adults.

OBJECTIVES/SPECIFIC AIMS: Cystic fibrosis (CF) is a life-shortening genetic disease that affects approximately 30,000 patients in the United States. CF patients suffer from chronic pulmonary infections that are associated with hyperinflammation and irreversible damage to the lower airways. As CF patients age, Pseudomonas aeruginosa (P.a.) is the predominant pathogen that infects the respiratory tract. The P.a. strains initially infecting the CF lung have a nonmucoid colony morphology, whereas, once chronic infection is established, these bacteria mutate leading to the emergence of mucoid P.a. variants with heightened resistance to both antibiotics and host immunity. Both nonmucoid and mucoid P.a. variants are often co-isolated on microbiological cultures of sputum collected from CF patients. However, the CF lung is known to exhibit heterogeneity in inflammation and infecting microbes across different lung regions that cannot be studied using routine sputum collection alone. Here, using a standardized bronchoscopic protocol, bronchoalveolar lavage (BAL) fluid was prospectively collected from each lobe of a CF cohort undergoing clinically indicated surgical procedures. We sought to investigate if there is an association between infecting P.a. variants (nonmucoid, mucoid, or mixed populations), the lung lobes in which these variants are found, and regional proinflammatory cytokine production. METHODS/STUDY POPULATION: We performed BAL on 16 CF patients with clinically stable disease. For each patient, we obtained BAL fluid from the right upper lobe, right middle lobe, right lower lobe, left upper lobe, lingula, and left lower lobe. We plated BAL fluid on nonselective and P.a.-selective medium to quantitate bacteria and to identify P.a. colony subtypes (nonmucoid, mucoid, or mixed). We further used a V-PLEX human cytokine array to quantitate inflammatory cytokine concentrations (IL-1β, TNF-α, IL-6, IL-8, and IL-10) within BAL fluid specimens. Our specimen collection was approved by the local IRB with informed consent and assent obtained from patient volunteers. RESULTS/ANTICIPATED RESULTS: Based on microbiological analysis, each lobar BAL specimen was classified as uninfected with P. a. or infected with nonmucoid, mucoid, or mixed (both nonmucoid and mucoid) P.a. variants. There was no observed propensity of mucoid or nonmucoid variants to be confined to certain lung lobes in our cohort. However, infection with mucoid P.a. variants was associated with higher concentrations of IL-1β (p<0.001), TNF-α (p<0.001), IL-8 (p<0.001), and IL-10 (p<0.001) within lobar BAL fluid compared with P.a.-free specimens. Specimens with mucoid variants also had greater concentrations of TNF-α (p<0.01), IL-8 (p<0.001), and IL-10 (p<0.05) compared with specimens with only nonmucoid P.a. variants. Patients infected with mixed mucoid and nonmucoid variants showed higher concentrations of TNF-α and IL-10 (p<0.05) as well as nonsignificant trends for higher concentrations of IL-1β and IL-6 compared to P.a.-free samples. Interestingly, the presence of nonmucoid P.a. variants was inversely correlated with IL-6 (p<0.05). Total bacterial burden (both P.a. and non-P.a. species) within BAL fluids was positively correlated with higher proinflammatory cytokine concentrations. Additionally, independent of bacterial colonization, the upper lobes (right upper lobe and left upper lobe) of the lungs showed trends towards higher proinflammatory cytokine concentrations compared with the lower lobes (right lower lobe and left lower lobe). DISCUSSION/SIGNIFICANCE OF IMPACT: Our results demonstrate that P.a. variants (mucoid or nonmucoid) appear not to be geographically restricted in ability to colonize any lobe of the CF lung. Moreover, infection with mucoid P.a. (either alone or in mixed populations with nonmucoid variants) is associated with higher inflammatory cytokine concentrations in the CF lung. Given that infection with mucoid P.a. predicts deterioration in pulmonary function, this study provides a rationale for further investigation of cytokines as diagnostic/prognostic correlates of infection and lung disease in CF.

OBJECTIVES/SPECIFIC AIMS: To create a searchable public registry of all Quality Improvement (QI) projects. To incentivize the medical professionals at UF Health to initiate quality improvement projects by reducing startup burden and providing a path to publishing results. To reduce the review effort performed by the internal review board on projects that are quality improvement Versus research. To foster publication of completed quality improvement projects. To assist the UF Health Sebastian Ferrero Office of Clinical Quality & Patient Safety in managing quality improvement across the hospital system. METHODS/STUDY POPULATION: This project used a variant of the spiral software development model and principles from the ADDIE instructional design process for the creation of a registry that is web based. To understand the current registration process and management of quality projects in the UF Health system a needs assessment was performed with the UF Health Sebastian Ferrero Office of Clinical Quality & Patient Safety to gather project requirements. Biweekly meetings were held between the Quality Improvement office and the Clinical and Translational Science – Informatics and Technology teams during the entire project. Our primary goal was to collect just enough information to answer the basic questions of who is doing which QI project, what department are they from, what are the most basic details about the type of project and who is involved. We also wanted to create incentive in the user group to try to find an existing project to join or to commit the details of their proposed new project to a data registry for others to find to reduce the amount of duplicate QI projects. We created a series of design templates for further customization and feature discovery. We then proceed with the development of the registry using a Python web development framework called Django, which is a technology that powers Pinterest and the Washington Post Web sites. The application is broken down into 2 main components (i) data input, where information is collected from clinical staff, Nurses, Pharmacists, Residents, and Doctors on what quality improvement projects they intend to complete and (ii) project registry, where completed or “registered” projects can be viewed and searched publicly. The registry consists of a quality investigator profile that lists contact information, expertise, and areas of interest. A dashboard allows for the creation and review of quality improvement projects. A search function enables certain quality project details to be publicly accessible to encourage collaboration. We developed the Registry Matching Algorithm which is based on the Jaccard similarity coefficient that uses quality project features to find similar quality projects. The algorithm allows for quality investigators to find existing or previous quality improvement projects to encourage collaboration and to reduce repeat projects. We also developed the QIPR Approver Algorithm that guides the investigator through a series of questions that allows an appropriate quality project to get approved to start without the need for human intervention. RESULTS/ANTICIPATED RESULTS: A product of this project is an open source software package that is freely available on GitHub for distribution to other health systems under the Apache 2.0 open source license. Adoption of the Quality Improvement Project Registry and promotion of it to the intended audience are important factors for the success of this registry. Thanks goes to the UW-Madison and their QI/Program Evaluation Self-Certification Tool (https://uwmadison.co1.qualtrics.com/SE/?SID=SV_3lVeNuKe8FhKc73) used as example and inspiration for this project. DISCUSSION/SIGNIFICANCE OF IMPACT: This registry was created to help understand the impact of improved management of quality projects in a hospital system. The ultimate result will be to reduce time to approve quality improvement projects, increase collaboration across the UF Health Hospital system, reduce redundancy of quality improvement projects and translate more projects into publications.

OBJECTIVES/GOALS: Serious video games are designed for skill-building and are increasingly being used for healthcare interventions with adolescents and young adults (AYAs). The study goal was to identify AYAs’ preferred game features, by demographic groups, to inform the development of a game to improve AYA’s engagement in their congenital heart disease (CHD) care. METHODS/STUDY POPULATION: Pediatric patients, 12-18 years old, completed surveys at a routine CHD care visit. Participants rated their likelihood of using games to learn CHD management skills (5-point Likert) and preferences for ten game features commonly used, such as: personalization (make your own avatar) and levels (unlock new, advanced stages as you do better). Participants selected one of three response options: 1=would make me less interested in the game, 2=doesn’t matter, 3=would make me more interested in the game. Descriptives and frequencies assessed interest in different game features. Chi-square tests were used to identify potential differences in game feature preferences by gender identity, age group (early/mid-adolescence vs. late adolescence), and race and ethnicity. RESULTS/ANTICIPATED RESULTS: Of 83 participants who completed surveys, the mean age was 15 years old (12-18; SD=1.73), 55% were male, 79% were Non-Hispanic White, and 70% were interested in video games for gaining CHD management skills. The top-rated game features were: levels (78%; unlock advanced stages), conflict (74%; face challenges), personalization (70%; create avatar), and story (70%; journey-based). The three lowest-ranked features were: time (29%; restricted time to complete challenge), competition (47%; score/play against others), strategy (53%; plan to reach goal). No significant differences in game feature preferences were found by demographic characteristics. DISCUSSION/SIGNIFICANCE: Most AYAs with CHD were interested in games, offering a promising avenue for future healthcare interventions. Given no significantly different preferences by demographics, the game may not require tailoring game features for certain groups. However, additional research with diverse participants is needed to fully inform game development.