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/Goals: The objective of this project was to develop electronic health record-based algorithms, or computational phenotypes, for carbapenem-resistant Enterobacterales (CRE) and extended-spectrum beta-lactamase-producing Enterobacterales (ESBL) infections based on public health surveillance case definitions. Methods/Study Population: We analyzed electronic health records (EHR) from a retrospective cohort of patients within the University of Colorado Health System. We detected CRE and ESBL infections using a set of string-matching algorithms for bacterial organism, specimen source, and antibiotic susceptibility test results. Infections were limited to the first case of each organism per patient in a 30-day period. We conducted manual chart review on a subset of cases and non-cases for validation purposes. We then performed a descriptive analysis to examine demographic characteristics of patients with CRE and ESBL infections including sex, age, infection type, and organism. All analyses were conducted using R statistical software. Results/Anticipated Results: There were 634 CRE infections from 2013 to 2023, with the majority from urine (n = 448, 70.7%) or blood specimens (n = 56, 8.8%). Over half of CRE case-patients were female (n =  362, 57.1%). The mean age was 65.0 years. Most isolates were identified as Enterobacter cloacae complex species (n = 353, 55.6%). There was a total of 4,363 ESBL infections from 2019 to 2023, and most were urine (n = 3,550, 81.4%) or blood specimens (n = 457, 10.5%). Most ESBL case-patients were female (n = 3,030, 69.4%) with a mean age of 61.2 years. The majority of isolates were identified as Escherichia coli (n = 2,932, 67.2%), followed by Klebsiella pneumoniae (n = 368, 8.4%). Validation results are pending, including sensitivity and specificity of the algorithms. Discussion/Significance of Impact: Demographic characteristics of CRE and ESBL case-patients were similar to findings from traditional public health surveillance. The algorithms will supplement traditional surveillance methods, inform future epidemiological research using EHR data, as well as contribute to the development of standardized EHR-based definitions for CRE and ESBL.

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: 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.

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.

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.

The Implementation Science Centers in Cancer Control (ISC3) initiative, funded by the National Cancer Institute, called for the development of implementation laboratories to bolster implementation science, create research-ready environments, and expedite adoption and implementation of evidence-based interventions (EBIs) into practice. The Building Research in Implementation and Dissemination to close Gaps and achieve Equity in Cancer Control (BRIDGE-C2) Center is one of seven ISC3 centers. BRIDGE-C2 aims to identify strategies to improve implementation of cancer prevention EBIs and conduct research / develop pragmatic methods to tailor, enhance, and support the adoption and sustainability of these strategies; advance implementation science; and build capacity and training opportunities. Since its inception, the BRIDGE-C2 Center has been conducting research and training activities to advance knowledge on how to effectively implement strategies to improve cancer prevention EBIs in primary care clinics serving socioeconomically disadvantaged patients. The translational science benefits model (TSBM) provides a useful framework for organizing a description of the BRIDGE-C2 Center’s activities. In this paper, we describe examples of BRIDGE-C2 activities and the specific impact indicators within each relevant domain/subdomain of the TSBM, demonstrating that a single activity or project has multiple impacts on methods and capacity building, clinical domains, and community health.

The pervasive problem of irreproducibility of preclinical research represents a substantial threat to the translation of CTSA-generated health interventions. Key stakeholders in the research process have proposed solutions to this challenge to encourage research practices that improve reproducibility. However, these proposals have had minimal impact, because they either 1. take place too late in the research process, 2. focus exclusively on the products of research instead of the processes of research, and/or 3. fail to take into account the driving incentives in the research enterprise. Because so much clinical and translational science is team-based, CTSA hubs have a unique opportunity to leverage Science of Team Science research to implement and support innovative, evidence-based, team-focused, reproducibility-enhancing activities at a project’s start, and across its evolution. Here, we describe the impact of irreproducibility on clinical and translational science, review its origins, and then describe stakeholders’ efforts to impact reproducibility, and why those efforts may not have the desired effect. Based on team-science best practices and principles of scientific integrity, we then propose ways for Translational Teams to build reproducible behaviors. We end with suggestions for how CTSAs can leverage team-based best practices and identify observable behaviors that indicate a culture of reproducible research.

Research articles in the clinical and translational science literature commonly use quantitative data to inform evaluation of interventions, learn about the etiology of disease, or develop methods for diagnostic testing or risk prediction of future events. The peer review process must evaluate the methodology used therein, including use of quantitative statistical methods. In this manuscript, we provide guidance for peer reviewers tasked with assessing quantitative methodology, intended to complement guidelines and recommendations that exist for manuscript authors. We describe components of clinical and translational science research manuscripts that require assessment including study design and hypothesis evaluation, sampling and data acquisition, interventions (for studies that include an intervention), measurement of data, statistical analysis methods, presentation of the study results, and interpretation of the study results. For each component, we describe what reviewers should look for and assess; how reviewers should provide helpful comments for fixable errors or omissions; and how reviewers should communicate uncorrectable and irreparable errors. We then discuss the critical concepts of transparency and acceptance/revision guidelines when communicating with responsible journal editors.

ABSTRACT IMPACT: We devised a new method to produce highly potent SARS-CoV2-specific that can be used to treat severely ill patients with Covid-19. OBJECTIVES/GOALS: Neutralizing antibodies against SARS-CoV-2 are thought to offer the most immediate and effective treatment for those severely afflicted by Covid-19. We devised an approach for rapid and efficient generation of human monoclonal antibodies with neutralizing activity against SARS-CoV-2. METHODS/STUDY POPULATION: SARS-CoV-2 S1 spike protein-specific memory B cells were isolated from 12 subjects recovering from infection with that virus. Paired end single index sequencing was performed using up to 10,000 antigen-specific B cells per subject. Antigen-specific B cell clones were identified by unique diversity and joining gene V(D)J rearrangements and the CDR3 regions. VH and VL regions were cloned and the products expressed in 293T/17 cells to generate spike-specific human monoclonal antibodies. RESULTS/ANTICIPATED RESULTS: Forty-three human monoclonal antibodies were produced. Every monoclonal antibody so generated neutralized viruses pseudotyped with Spike protein of the Wuhan-1 strain. Eighteen monoclonal antibodies neutralized pseudotyped viruses with half-maximal inhibitory concentration (IC50s) between 1 pg/mL and 1 ng/mL (6.7 x 10E-15 M to 6.7 x 10E-12 M), exceeding by 10-100-fold the potency of previously reported anti-SARS-CoV-2-neutralizing monoclonal antibodies. Eight monoclonal antibodies neutralized viruses pseudotyped with mutant spike proteins previously identified in clinical isolates, including receptor binding domain mutants and the C-terminal D614G mutant with IC50<6.7 x10E-12M. DISCUSSION/SIGNIFICANCE OF FINDINGS: We show that SARS-CoV-2 evokes high affinity B cell responses. Some B cells produce antibodies that are broadly neutralizing; others produce strain-specific antibodies. However, antigenic variants that would potentially escape control by immunity or vaccination were nonetheless identified.

OBJECTIVES/SPECIFIC AIMS: Parkinson’s disease (PD) is a condition that affects over a million Americans, and despite current medical therapies, the progression of the disease results in impaired generation of internally timed or guided (IG) movements. To address this loss of motor function, previous rehabilitation therapies have focused on remediating the affected striatal-thalamic-cortical circuits (STC), primarily thought to be responsible in generating timed motor patterns. However, given the disease leads to the cell death of dopaminergic cells that are essential for proper STC function, we propose a motor therapy aimed at utilizing a compensatory parallel cerebellar-thalamic-cortical (CTC) pathway, recruited to perform externally guided (EG) movements, in which gait initiation is driven from sensory input. Our previous study has shown efficacy in our novel argentine tango therapy and improves behavioral measures above the relevant MCID threshold, but it has not been established that the CTC are in the causal pathway that are responsible for these changes. Using neural measures from task fMRI, we have begun to characterize networks that have changed and quantify any associations with behavioral metrics. METHODS/STUDY POPULATION: Patients were randomly assigned to an IG (n=18), EG (n=18), or education contact control (n=14). Participants were assessed preintervention and postintervention for behavioral motor and cognitive measures and neurophysiologically with task based fMRI. In the task, participants performed a foot tapping task under both IG (tap their foot in previously learned rhythm) or EG (tap immediately after receiving a tactile cue on their hand) conditions. The fMRI data were preprocessed using AFNI and registered to MNI standard space. The brainnetome atlas was applied and the average time series of each region of interest (ROI) was used to increase the signal to noise ratio. The activation of these ROI with respect to the stimulus was modeled using GLM, and we estimated the area under the curve during the task blocks. A 1-way ANOVA analysis on these betas were performed between the pre and the post intervention time points and the ROIs that were above a significance of 0.95 were identified and corrected for multiple comparisons. The change in beta in all ROIs for each individual were calculated and then correlated with the changes in the behavioral data, to see which changes in ROI areas matched the best with the behavioral changes. RESULTS/ANTICIPATED RESULTS: The EG group showed significant changes only in the EG task in 2 areas—inferior frontal gyrus and inferior temporal sulcus. Correlating to the cognitive behavioral measures show reduced error from the Inferior frontal gyrus (corr>0.5) best reflect changes in observed. There were no changes to either the STC or the CTC pathways. The IG group showed no changes behaviorally and showed no changes neurally as well. The control group showed no changes behaviorally, but neuronally certain DMN nodes, such as the precuneus and inferior temporal regions showed a significant change for both tasks. DISCUSSION/SIGNIFICANCE OF IMPACT: Addressing the damaged STC pathway directly through IG therapy may not be effective. The EG therapy may not be able to enhance the STC pathway. However, the therapy appears to utilize new areas in the frontal regions and correlates with positively with changes in spatial memory and balance tasks. Contrary to our hypothesis the CTC circuit was not upregulated for performance of the IG or EG task, but therapy may have enhanced recruitment of other cognitively engaged areas. The educational control group interestingly showed changes in the DMN network, which has been shown to be linked to attention during tasks blocks.

ABSTRACT IMPACT: Development of our animal model of moyamoya will provide a meaningful assessment of therapeutic efficacy of interventions applicable to the clinical setting. OBJECTIVES/GOALS: Moyamoya is a cerebrovascular condition with progressive stenosis of the internal carotid arteries (ICA) and formation of abnormal vascular collaterals at the base of the brain, all of which result in ischemic and hemorrhagic strokes. We aim to develop a needed animal model of this condition in order to develop new therapeutics. METHODS/STUDY POPULATION: Male and female C57Bl/6J mice (4 months old) underwent surgery for the unilateral placement of a microcoil (0.16 mm ID) onto the proximal ICA or sham control. After 30 and 60 days (N = 6-8/time point), the brain blood vessels were examined for changes in diameter, number of anastomoses, and development of new collaterals using DiI stain. Brain tissue was examined for micro-hemorrhages using Prussian blue stain, and cross-sections of blood vessels were examined for intimal thickening using H&E and smooth muscle actin. Expression of vascular endothelial growth factor (VEGF), which is associated with angiogenesis and moyamoya syndrome, was quantified by qPCR. Blood samples were also analyzed for inflammatory biomarkers using ELISA. RESULTS/ANTICIPATED RESULTS: Within 30 days, the distal ICA and anterior cerebral artery (ACA) had significantly decreased diameters at the Circle of Willis, with an initial decrease in the number of cortical anastomoses. Histology demonstrated smaller lumen diameter and alterations to in the various layers of the blood vessels, indicating intimal thickening and stenosis of the affected blood vessels. There was also a significant increase in the number of intracranial micro-bleeds, suggesting a compromised vascular integrity. This may be due, in part, to a significant upregulation in VEGF gene expression within the striatum, a region of hemorrhagic occurrence in moyamoya patients. DISCUSSION/SIGNIFICANCE OF FINDINGS: We report the development of an animal model with vasculopathies that mimic those observed in patients with moyamoya syndrome. With further characterization, this animal model will have a positive impact as a meaningful assessment of therapeutic efficacy of interventions applicable to the clinical setting.

OBJECTIVES/GOALS: Aim 1: To explore methodologies to identify gender minorities, and Filipinos, separately, in an electronic health record system. Aim 2: To characterize the similarities and differences in demographic, socioeconomic, and mental health of Filipinos gender minorities compared to Native Hawaiian Pacific Islanders, and White/ European Americans. METHODS/STUDY POPULATION: This study was approved by the University of Hawaii Institutional Review Board. Cross-sectional retrospective data were obtained from a collaborative community clinic’s electronic health record system. Patients were age 18 and older with a clinical diagnosis for gender dysphoria and from Native Hawaiian Pacific Islander, White/ European American, and Filipino backgrounds. RESULTS/ANTICIPATED RESULTS: Preliminary data revealed that 11% of the clinical population were diagnosed with gender dysphoria (N=373) with 57.6% (n=215) who met the inclusion criteria with complete health registration forms. Patients were from Filipino (21.8%), Native Hawaiian Pacific Islander (23.3%), White/ European American (31.6%), and multiethnic (23.3%) backgrounds. Most patients reported mental health (e.g., depression) conditions (50.6%-64.7%). Further statistical analyses will reveal if Filipinos have higher or lower levels of anxiety, depression, and suicide risks than Native Hawaiian Pacific Islander, and White/ European American gender minority individuals. DISCUSSION/SIGNIFICANCE: In Hawaii, one person dies by suicide every two days; suicide is the lead cause of fatal injuries. Study findings can inform future methodology studies to identify gender minorities and develop culturally relevant gender affirming mental health programs for gender minorities.

OBJECTIVES/SPECIFIC AIMS: The objective of this project is the application of complex fusion models, which combine observed and modeled data, to areas with sparse monitoring networks with multiple chemical components is under-developed. Such models could provide improved accuracy and coverage for air quality measurement predictions, an area greatly limited by the amount of missing data. METHODS/STUDY POPULATION: This project focuses on the development of methods for improved estimation of pollutant concentrations when only sparse monitor networks are found. Sparse monitoring networks are defined as areas where fewer than three criteria air pollutants (based on EPA standards) are monitored. Particularly, a multivariate air pollutant statistical model to predict spatio-temporally resolved concentration fields for multiple pollutants simultaneously is developed and evaluated. The multivariate predictions allow monitored pollutants to inform the prediction of nonmonitored pollutants in sparse networks. RESULTS/ANTICIPATED RESULTS: Daily, ZIP code level pollutant concentration estimates will be provided for 8 pollutants across South Carolina, and goodness of fit metrics for model variants and previously established methods will be compared. DISCUSSION/SIGNIFICANCE OF IMPACT: These methods utilize only widely available data resources, meaning that the improved predictive accuracy of sparsely monitored pollutant concentrations can benefit future studies in any US area by improving estimation of health effects and saving resources needed for supplemental air pollutant monitoring campaigns. Our method for estimation attempts to improve predictive accuracy and data availability for sparsely monitored pollutants and areas.