OBJECTIVES/GOALS: To explore the severity of posttraumatic stress disorder (PTSD) symptoms in association with hippocampal and amygdala volumes in ICU survivors. We hypothesize that the severity of posttraumatic stress symptoms in ICU survivors is associated with lower volumes of both the hippocampus and amygdala. METHODS/STUDY POPULATION: Secondary analysis of the VISIONS study, a prospective sub-study of the BRAIN-ICU cohort, which included survivors of critical illness. Patients were screened for preexisting PTSD before discharge. The PTSD Checklist Specific (PCL-S) was used at 3 and 12 months to evaluate the ICU as a traumatic experience. A score of >30, indicated significant symptoms of PTSD. A Philips Achieva 3T MRI scanner was used to scan patients at both discharge and 3-month follow-up. To compare median brain volumes at discharge and 3 months for those with and without significant PTSD symptomatology (PCL-S ≥30) at 3 and 12 months, we used a Kruskal-Wallis (KW) equality-of-populations rank test. RESULTS/ANTICIPATED RESULTS: The median age for our sample was 58.5 (52.6, 63.7). One-third of the sample was female, and 90% were Caucasian. Fifty-seven percent of individuals (N = 12) had at least one prior mental health diagnosis, with two having a prior history of PTSD. One third of individuals experienced delirium during their critical illness. At 3-month follow up, there were three patients with PTSD symptomatology and one at 12-month follow up. Median brain volumes (hippocampus or amygdala) did not differ between individuals with or without PTSD symptomatology at either 3 or 12 months (p-values for all tests >0.05). DISCUSSION/SIGNIFICANCE OF IMPACT: Although our study did not reveal significant differences in brain volumes between PTSD patients and non-PTSD patients, sample size is a major limitation and larger scale studies should be undertaken to elucidate possible neurobiological markers of PTSD in ICU survivors. CONFLICT OF INTEREST DESCRIPTION: Dr. Wilson would like to acknowledge salary support from the Vanderbilt Faculty Research Scholars Program (1KL2TR002245), HL111111 and GM120484. Drs. Ely and Jackson as well as Mrs. Kiehl all receive funding for their time working on this investigation from AG035117 and HL111111. Dr. Ely would additionally like to acknowledge salary support from the Tennessee Valley Healthcare System Geriatric Research Education and Clinical Center (GRECC). Dr. Ely will also disclose additional funding for his time from AG027472 and having received honoraria from Orion and Hospira for CME activity; he does not hold stock or consultant relationships with those companies. The authors would like to acknowledge the following: this work was conducted in part using the resources of the Center for Computational Imaging at Vanderbilt University Institute of Imaging Science and the Advanced Computing Center for Research and Education at Vanderbilt University, Nashville, TN, and study data were collected and managed using REDCap electronic data capture tools hosted at Vanderbilt University.

OBJECTIVES/GOALS: To reduce hospitalizations, health care systems are studying ways of improving social determinants of health (SDoH) in patients with chronic disease such as diabetes (DM). Our goal was to better characterize the SDoH of a cohort of DM patients by using socio-economic information from census data. METHODS/STUDY POPULATION: Our study population included DM patients seen in primary care practices of a large health care system in 2013-2017. We integrated socio-economic status (SES) information from the American Factfinder to data extracted from the electronic health record (EHR). Addresses for the cohort were geocoded using ArcMap to obtain the census tract information for median income, poverty status, educational level, and supplemental food benefits using American Community Survey 5-Year estimates. We used multivariable logistic regression to calculate odds ratio (OR) and 95% confidence intervals [], with 3+ comorbidities as the dependent variable and demographic and SES variables as independent variables. RESULTS/ANTICIPATED RESULTS: Our study population included 13,782 patients: 53% were female, 65% white, 28% Black, 27% were on Medicare, 3% on Medicaid, median age was 60, 53% had 3+ comorbidities. Median income was $66,243, poverty level 6%, receiving food benefits 8%, no high school degree 8%, and bachelor’s degree or higher 30%. After evaluating collinearity, our multivariable analysis showed that patients with 3+ comorbidities were more likely to have income < $52,000 (lower quartile) versus $84,001 (upper quartile), OR = 1.2 [1.0-1.4]; be female, OR = 1.6 [1.4-1.7]; divorced or widowed versus married, OR = 1.5 [1.3-1.7], 1.4 [1.3-1.6]; and be on Medicare, Medicaid or both, OR = 2.4 [2.2-2.6], 2.2 [1.8-2.6], 6.0 [4.5-8.3]. DISCUSSION/SIGNIFICANCE OF IMPACT: Census tract-based SES could provide invaluable information to health care providers when associated to the EHR. We found that median income, which is not collected in the EHR, was significantly associated with a higher burden of disease. Census tract SES could serve as a proxy for evaluating SDoH.

ABSTRACT IMPACT: Neighborhood disadvantage was significantly associated with brain structure and function in trauma-exposed adults, providing evidence that contextual factors should be assessed in mental health research, particularly in high-risk populations. OBJECTIVES/GOALS: Over 13 percent of Americans live in a socioeconomically disadvantaged neighborhood. Previous work has linked lower individual socioeconomic position to alterations in brain structure and function. However, the neural effects of area-level socioeconomic factors, such as neighborhood disadvantage, are unclear. METHODS/STUDY POPULATION: We recruited two-hundred and fifteen traumatically-injured participants from an Emergency Department in southeastern Wisconsin. An Area Deprivation Index (ADI) score, a national measure of neighborhood socioeconomic disadvantage, was derived from each participant’s home address. Two-weeks post-trauma, participants underwent a battery of self-report measures and functional magnetic resonance imaging (fMRI) scans. Using a multi-modal approach, we investigated the impact of ADI on brain structure as well as neural activation during rest and during an emotional uncertainty task. We sought to disentangle the relationship between neighborhood and individual socioeconomic position and neural activity in the context of trauma. RESULTS/ANTICIPATED RESULTS: We demonstrated that neighborhood disadvantage is associated with decreased volume and alterations of resting state functional connectivity of structures implicated in affect processing, including the hippocampus, amygdala, and ventromedial prefrontal cortex. These results held even after controlling for relevant individual variables, including acute post-traumatic stress symptoms and years of education. Moreover, individuals from disadvantaged neighborhoods exhibited heighted activation of these same structures in response to aversive stimuli. Thus, brain regions critical for recognizing and processing negative stimuli are susceptible to the effects of area-level socioeconomic factors. DISCUSSION/SIGNIFICANCE OF FINDINGS: The results offer additional evidence that neurobiological mechanisms clarify how stress ‘gets under the skin’. Changes to key brain regions may explain why those living in disadvantaged neighborhoods are at a heighted risk of PTSD. Broadly, these findings should inform future policies and community-driven interventions aimed at reducing poverty.

OBJECTIVES/SPECIFIC AIMS: This study aims to first describe the unique cytokine profile and TGFbeta levels of young children with CF, then understand the pathologic effects of TGFbeta on lung function in a CF animal model. These powerful translational studies linking observations in clinical disease with a transgenic mouse model allow us a unique opportunity to investigate the role of TGFbeta in early CF lung disease. METHODS/STUDY POPULATION: Cytokine levels (TGFbeta, TNFalpha, IL-8, IL-6, HNE, and IL-1beta) in bronchoalveolar lavage fluid (BALF) from CF patients (n = 15) and non-CF control patients (n = 21) under 6 years old were determined by ELISA and Luminex assay. Tracheotomized patients without significant underlying lung disease were chosen as non-CF inflamed control patients, as they had similar levels of neutrophilic inflammation and infection as CF patients. The percentage of BAL neutrophils (% PMNs) in each sample was assessed. The relationships between cytokines were analyzed using linear regression and principal components analysis. In animal studies, CF and non-CF mice (n = 4-5 per group) were treated with intratracheal adenoviral TGFbeta1 vector, an empty vector control, or PBS. After one week, animals were collected; lung function, response to the bronchoconstrictor methacholine, and rescue with albuterol were measured utilizing a FlexiVent machine. Lungs were collected for histology. Immunohistochemistry for alpha-SMA was performed and pictures of all cross-sectional airways were obtained. Burden of ASM was assessed by dividing the square root of alpha-SMA stained airway smooth muscle by the basement membrane perimeter length of each airway. RESULTS/ANTICIPATED RESULTS: Patient characteristics of CF and non-CF inflamed control patients were similar in terms of age (3.6 yrs vs 3.3 yrs respectively, p = 0.49), positive BAL culture (13% vs 14%, p = 0.94), and % PMNs (65% vs 56%, p = 0.64). Despite these similarities, TGFbeta levels were 2-fold higher in CF versus non-CF BAL (p = 0.034). Analysis of BAL cytokines from both patient groups showed that three principal components describe 86% of total variance across the cytokine variables. These components represent different contributions from the cytokines, with TGFbeta, IL6, and % PMNs comprising one component of similarly regulated inflammatory markers. These components can distinguish CF versus non-CF patients with 77% accuracy (area under the curve: 0.77). TGFbeta concentrations were uniquely associated with increased IL-6 in CF samples (r = 0.74; p = 0.0015) but did not demonstrate association with other cytokines. After TGFbeta exposure, CF mice demonstrated greater abnormalities in airway resistance than non-CF mice, with heightened response to methacholine. Importantly, this increase in airway obstruction in CF mice was reversible with albuterol treatment, indicating airway smooth muscle dysfunction as a principal driver of lung function abnormalities. Furthermore, TGFbeta induced an increased ASM burden on lung histology in both CF and non-CF mice (p<0.05). IL-6 levels in the BAL of CF mice showed greater increases after TGFbeta treatment compared to non-CF mice (p<0.05). Empty vector control treatment did not cause lung pathology. DISCUSSION/SIGNIFICANCE OF IMPACT: Young children with CF have a unique pattern of pulmonary inflammation compared to inflamed non-CF control patients. In CF, TGFbeta pulmonary levels are uniquely associated with IL-6, a driver of ASM dysfunction in other pulmonary diseases. We followed up this clinical observation study by investigating the effect of TGFbeta on pulmonary disease in a mouse model. CF mice demonstrate increased pulmonary IL-6, airway obstruction, and ASM dysfunction after TGFbeta exposure. This study provides evidence that TGFbeta is associated with a distinct cytokine pattern that may promote ASM dysfunction in early CF lung disease. Understanding the mechanism of early CF pathophysiology will be critical in developing targeted therapeutics that can prevent early lung damage.

OBJECTIVES/GOALS: Recent NCATS funding announcements emphasize pursuing domain-agnostic translational science projects that seek to transform the system of science. We aimed to articulate the social responsibility of translational science, defined as prioritizing improved health outcomes and decreased disparities. METHODS/STUDY POPULATION: We focused on the framing of social responsibilities of translational science and distinctions between (a) domain-agnostic translational science that aims to transform the system of science and (b) translational research that takes place within a specific therapeutic area. We reviewed CTSA funding calls, translational research ethics papers, and statements by leaders in the field of translational science. We integrated the social responsibilities of improving health outcomes and decreasing disparities with the values of translational science, which prioritize the relevance, usability, and sustainability of translational interventions. RESULTS/ANTICIPATED RESULTS: We drew on our review of the literature and case studies to offer guidance aimed at helping to ensure that differently positioned actors and entities within the translational ecology can advance the values of translational science while also fulfilling the social responsibilities of translational science. We specify how (a) Funders and policymaking institutions, (b) Organizations such as research universities and CTSA institutes, (c) Translational health science teams working on innovative translational science projects, and (d) Individual translational scientists can all contribute to ensuring that translational science fulfills its ethical obligations and social responsibilities. DISCUSSION/SIGNIFICANCE: The social responsibility of translational science can be fulfilled by centering its efforts to develop useful, sustainable, and relevant innovations. These criteria clarify how social responsibilities manifest in practice and can help funders shape and guide the next era of translational discovery.

OBJECTIVES/SPECIFIC AIMS: The objective of this research is to determine under what conditions endpoints based on estimated glomerular filtration rate (eGFR) slope or on relatively small declines in eGFR provide valid and useful surrogate endpoints for pivotal clinical trials in chronic kidney disease (CKD) patients. METHODS/STUDY POPULATION: We consider 2 classes of surrogate endpoints. The first class includes endpoints defined by the average rate of change in eGFR during defined portions of the follow-up period of the trial, following initiation of the randomized treatment interventions. The second class includes composite endpoints defined by the time from randomization until the occurrence of a designated decline in eGFR or kidney failure. The true clinical endpoint is considered to be the time from randomization until kidney failure, irrespective of the trajectory in eGFR measurements prior to kidney failure. We apply statistical simulation to determine conditions under which alternative endpoints within the 2 classes are (1) valid surrogate endpoints, in the sense of preserving a low probability of rejecting the null hypothesis of no treatment effect on the surrogate endpoint when there is no treatment effect on the clinical endpoints and are also (2) useful surrogate endpoints, in the sense of providing increased statistical power that allows significant reductions in sample size and/or duration of follow-up. Input parameters for the simulations include (a) characteristics of the joint distribution of the longitudinal eGFR measurements and the time to occurrence of renal failure, (b) characteristics of the short-term and long-term effects of the treatment, and (c) design parameters, including the duration of accrual and follow-up and the spacing of eGFR measurements during the follow-up period. We use joint analyses of 19 treatment comparisons across 13 previous clinical trials of CKD patients to guide the selection of input parameters for the simulations. We apply longitudinal mixed effects models for analysis of endpoints based on eGFR slope, and Cox regression for analyses of the composite time-to-event endpoints. RESULTS/ANTICIPATED RESULTS: We have previously shown that surrogate endpoints defined by eGFR declines of 30% or 40% can provide valid and useful alternative endpoints in CKD clinical trials for interventions that do not produce short-term effects on eGFR which differ from the longer-term effects of the interventions. Other factors influencing the validity and utility of these endpoints include the average baseline eGFR, the mean rate of change in eGFR, and the extent to which the size of the treatment effect depends on the patient’s underling rate of eGFR decline. We will extend these results by presenting preliminary results describing conditions under which outcomes based on eGFR slope provide valid and useful alternatives to the clinical endpoint of time until occurrence of kidney failure. DISCUSSION/SIGNIFICANCE OF IMPACT: The statistical simulation strategy described in this research can be used during the design of clinical trials of chronic kidney disease to assist in the selection of endpoints that maximize savings in sample size and duration of follow-up while retaining a low risk of producing a false positive conclusion in the absence of a true effect of the treatment on the time until kidney failure.

Objectives/Goals: This project investigates how early childhood neighborhood factors influence attention-deficit/hyperactivity disorder (ADHD) outcomes in adolescence. Poor neighborhood conditions have been linked to higher ADHD rates; however, the effects of these factors on academic achievement, social relationships, and risk-taking behaviors remain understudied. Methods/Study Population: A large, diverse, harmonized, cleaned dataset from a national multisite research program (Environmental Influences on Child Health Outcomes (ECHO)) will be used. Neighborhood factors will be measured using geocoded, census-level indices of neighborhood quality: the Child Opportunity Index 3.0. Adolescent outcomes include self and caregiver-reported measures of comorbid psychopathology, risk-taking behavior, and academic and social functioning. A series of regression analyses will be conducted to examine the relationship between these variables. An estimated 6000 children are expected to be included in the analyses. Results/Anticipated Results: We expect that poorer neighborhood conditions, particularly low social and economic resources, will be associated with lower overall functioning in adolescence, and that this relationship will be stronger among adolescents with ADHD relative to those without ADHD. Discussion/Significance of Impact: By identifying risk and protective factors, this project will help identify potential prevention and treatment targets for a substantial number of youth and may inform policy efforts to improve resource equity and reduce existing disparities.

OBJECTIVES/GOALS: Last year, we reported on storytelling events highlighting the experiences of Black biomedical professionals. The goal of this continuation study was to put Black Voices into action through a companion curriculum, instructional materials, and facilitated meaningful discussions around racial justice. METHODS/STUDY POPULATION: We developed instructional materials and pilot-implemented the curriculum in a 250-student Healthcare Leadership class. Prior to viewing the video, participants were asked to provide a definition of diversity, inclusion, and equity. Afterwards, the students reflected and suggested changes to the definition. Throughout the class, instructors facilitated group discussions about the role of diversity. As a culminating activity, students submitted a written reflection summarizing their understanding of the role of diversity, inclusion, and equity in healthcare. RESULTS/ANTICIPATED RESULTS: Instructional materials included videos, a facilitators guide, a reflection worksheet, and online course modules. The facilitators guide included an overview of best practices in the facilitation of group discussions, common rules groups can establish for open dialogue, and discussion questions specific to the Black Voices videos. In-class and lab discussions brought up several prominent metaphors for diversity and inclusion. For example, students first defined diversity as pieces of the puzzle that somehow fit together, but later surmised that the pieces do not need to fit, or be from the same puzzle. DISCUSSION/SIGNIFICANCE: Our pilot showed that participants experienced the power of storytelling, particularly for amplifying Black voices, gained a vehicle to expand awareness and consciousness of diversity, equity, and inclusion, and identified plans to promote and facilitate stories of race, equity, and diversity.

OBJECTIVES/SPECIFIC AIMS: Diabetic retinopathy is an increasingly prevalent disease, difficult to screen for across the globe. We have developed and began optimizing an innovative technique to visualize and quantify retinal blood flow, to elucidate the role of the choroid in retinal pathologies such as diabetic retinopathy or choroidopathy. METHODS/STUDY POPULATION: Preliminary retinal was obtained from a surgical retina video library (Truvision, Goleta, CA, USA). Videos of different organs were recorded while vessels were occluded via a blood pressure cuff, using consumer-grade digital video cameras (NEX-5T, a7sii; Sony, New York, NY, USA). All other retinal videos were taken using a fundus camera (50×; Topcon, Oxland, NJ, USA) modified to support the above digital video cameras. All videos were processed using experimental software (MATLAB, Mathworks, Natick, MA, USA). RESULTS/ANTICIPATED RESULTS: Video imaging of the retina was optimized for lighting conditions and software requirements. Parameters were defined for the software imaging pipeline, such as frequency range of interest, sampling rate, and noise minimization. Software was developed to stabilize frames, accounting for eye saccades. Use of a biosensor enabled accurate measurement of pulse waveform, increasing signal-to-noise ratio. The optimal light requirements were determined such that adequate exposure of the retina is reproducible yet still comfortable for use in human subjects. DISCUSSION/SIGNIFICANCE OF IMPACT: This novel technique allows for an inexpensive, noninvasive, and reproducible ocular blood flow imaging platform. By optimizing this technique, we can proceed with our future plans for a pilot study to compare our imaging technique with the current standard, paving the way for future clinical studies.

OBJECTIVES/GOALS: Developing pharmacokinetic (PK) models to guide selective serotonin reuptake inhibitor (SSRI) dosing in youth is costly, time-intensive, and requires large numbers of participants. We evaluated the use of remnant blood samples from SSRI-treated youth and developed precision PK dosing strategies. METHODS/STUDY POPULATION: Following IRB approval, we used a clinical surveillance platform to identify patients with routine phlebotomy within 24 hours of escitalopram or sertraline dosing. Remnant blood samples were obtained from youth aged 5–18 years, escitalopram and sertraline concentrations were determined, and clinical characteristics (e.g., age, sex, weight, concomitant medications that inhibit sertraline or escitalopram metabolism) and phenotypes for CYP2C19, the predominant enzyme that metabolizes these SSRIs, were extracted from the electronic medical record (EMR). A population PK analysis of escitalopram and sertraline was performed using NONMEM. The influence of clinical variables, CYP2C19, and dosing was evaluated from simulated concentration-time curves. RESULTS/ANTICIPATED RESULTS: Over 21 months, we collected315 samples from escitalopram-treated patients (N=288) and 265 samples from sertraline-treated patients (N=255). In youth, escitalopram and sertraline exposure (concentrations over time) and specific pharmacokinetic parameters (e.g., clearance) were influenced by CYP2C19 phenotype, concomitant CYP2C19 inhibitors, and patient-specific characteristics. Escitalopram and sertraline concentrations from remnant blood samples were 3.98-fold higher and 3.23-fold higher, respectively, in poor metabolizers compared to normal metabolizers (escitalopram, p<0.001) and compared to normal, rapid, and ultrarapid metabolizers combined (sertraline, p<0.001). DISCUSSION/SIGNIFICANCE: Combining remnant blood sampling with pharmacogenetic-integrated EMR data can facilitate large-scale population PK analyses of escitalopram and sertraline in youth. This real-world approach can be used to rapidly develop precision SSRI dosing strategies, including slower titration and reduced target doses in CYP2C19 poor metabolizers.

Effectively addressing public health crises requires dynamic and nimble interdisciplinary collaborations across the translational spectrum, from bench to clinic to community. The Clinical and Translational Science Award (CTSA) Program hubs are uniquely suited to facilitate interdisciplinary collaborations across universities and academic medical centers. This paper describes the activities at the Columbia University CTSA Program hub to address a current public health crisis, the opioid epidemic. Columbia’s CTSA Program hub led a three-phase approach, based on the Conceptual Model of Transdisciplinary Scientific Collaboration as described by Stokols et al.: (1) a university-wide planning and brainstorming phase to identify key leaders across many domains who are influential in addressing the opioid epidemic, (2) a campus-wide and community outreach to identify all interested parties, and (3) ongoing targeted support for collaboration development. Preliminary metrics of success are interdisciplinary collaborations and grant funding. We describe recent examples of how interdisciplinary collaboration, academic-community partnership, and pilot funding contributed to the development and funding of innovative interdisciplinary research, including the New York site of the HEALing Communities initiative. The processes are now being used to support interdisciplinary approaches for other translational public health issues.

The critical processes driving successful research translation remain understudied. We describe a mixed-method case study protocol for analyzing translational research that has led to the successful development and implementation of innovative health interventions. An overarching goal of these case studies is to describe systematically the chain of events between basic, fundamental scientific discoveries and the adoption of evidence-based health applications, including description of varied, long-term impacts. The case study approach isolates many of the key factors that enable the successful translation of research into practice and provides compelling evidence connecting the intervention to measurable changes in health and medical practice, public health outcomes, and other broader societal impacts. The goal of disseminating this protocol is to systematize a rigorous approach, which can enhance reproducibility, promote the development of a large collection of comparable studies, and enable cross-case analyses. This approach, an application of the “science of translational science,” will lead to a better understanding of key research process markers, timelines, and potential points of leverage for intervention that may help facilitate decisions, processes, and policies to speed the sustainable translational process. Case studies are effective communication vehicles to demonstrate both accountability and the impacts of the public’s investment in research.

OBJECTIVES/SPECIFIC AIMS: The objective of this study is to develop patient-derived “personalized” organotypic models of human distal airways, in which basal stem cells (BCs) isolated from the pre-/terminal conducting airway region are co-cultured with autologous stromal cells from the same region to reproduce patient-specific distal airway epithelial-stromal units and their remodeling in COPD. METHODS/STUDY POPULATION: We established a protocol to isolate and propagate epithelial BCs, fibroblasts, and endothelial cells from the distal airways of normal and COPD lung donors. Heterogeneous cellular and molecular phenotypes in the human distal airways were characterized using immunofluorescence and single-cell RNA sequencing. Patient-specific distal airway epithelial-stromal units were reconstructed by co-culturing BCs and autologous stromal cells using an air-liquid interface-based airway wall model and a bronchosphere-based 3D distal airway organoid assay. RESULTS/ANTICIPATED RESULTS: Histologic analysis of derived epithelial-stromal units revealed heterogeneous patient-specific phenotypes characterized by hypo-/hyper-/metaplastic lesions (hypo-regenerative phenotype, mucous cell hyperplasia, squamous metaplasia, distal-to-proximal repatterning) in the epithelial compartment, accompanied, in some samples, by stromal remodeling. Candidate epithelial-stromal cross-talk mechanisms were identified using quantitative real-time RT-PCR analysis of autologous epithelial and stromal compartments of established patient-specific distal airway unit models. DISCUSSION/SIGNIFICANCE OF IMPACT: Epithelial and stromal cells isolated from distal airways of subjects with and without COPD can be assembled into functional, organ-level tissue which mimics the architecture of human distal airways and, in patients with COPD, reproduces several distal airway remodeling phenotypes. Patient-specific models of distal airway epithelial-stromal cross-talk established in this study can be used to identify candidate pathways that mediate disease-relevant airway remodeling and potentially utilized as pre-clinical platforms for developing personalized therapeutic approaches to suppress the progression of distal airway remodeling in chronic lung diseases, including COPD.

ABSTRACT IMPACT: This research takes a transcriptomic approach to parse genes and molecular pathways that underlie the fear memory circuitry and, in doing so, identifies therapeutic targets that can further be developed into treatments for fear disorders, such as post-traumatic stress disorder. OBJECTIVES/GOALS: Normal fear learning produces avoidance behavior that promotes survival, but excessive and persistent fear after trauma can lead to development of phobias and post-traumatic stress disorder (PTSD). Our goal is to understand the mechanism and identify novel genetic targets underlying fear responses. METHODS/STUDY POPULATION: Involvement of the basolateral amygdala (BLA) in fear acquisition is well established and requires activation of N-methyl-D-aspartic acid receptors (NMDARs). At a cellular level, NMDAR activation leads to production of nitric oxide (NO) by a process mediated by interaction between postsynaptic density protein 95 (PSD95) and neuronal nitric oxide synthase (nNOS). To elucidate mechanisms underlying the role of the PSD95-nNOS-NO pathway in conditioned fear, here we use rodent behavioral paradigms, pharmacological treatment with a small molecule PSD95-nNOS inhibitor, RNA-sequencing, and an AAV-mediated knockdown of the nNOS gene in the BLA. RESULTS/ANTICIPATED RESULTS: We show that treatment with ZL006 attenuates rodent cued-fear consolidation. Additionally, with RNA-sequencing, expression of 516 genes was altered in the BLA following fear expression; of these genes, 83 were restored with systemic ZL006 treatment. Network data and gene ontology enrichment analyses further revealed that cGMP effects, insulin-like growth factor binding, and cognition-related pathways were significantly altered. Finally, we show that a BLA-specific knockdown of nNOS attenuates cued fear consolidation, without adverse effects on other memory and motor behaviors. DISCUSSION/SIGNIFICANCE OF FINDINGS: Via a model of NMDAR-mediated fear consolidation, our results reveal novel pathways and genetic targets that underlie plasticity of fear memory circuitry. Importantly, these results will inform future therapeutic strategies for targeting fear related disorders like PTSD.

OBJECTIVES/GOALS: The objective of this study is to assess the feasibility and preliminary impact of a physiotherapy protocol for developing an individualized home-based physical activity program to increase physical activity (PA) levels in sedentary older adults with Type II Diabetes Mellitus (T2DM) living in Puerto Rico (PR). METHODS/STUDY POPULATION: This will be a pilot study with two phases. In phase 1, we will design a novel patient-centered home-based PA program protocol for adults ≥65 years with T2DM based on the Information-Motivation-Behavioral Skills model. Its content validity will be assessed through focus groups with 10 experts and 10 older adults and analyzed using a directed content analysis. Phase 2 we will be program implementation using a one-group, repeated measures design with 12 adults ≥65 years with T2DM. PA levels will be assessed by recording active minutes with a Fitbit. Risk of falls, balance, strength, and physical function will be assessed through standardized tests validated for this population. Statistical analysis will include descriptive statistics, comparisons via chi-square/Fisher’s exact test, and non-parametric tests. RESULTS/ANTICIPATED RESULTS: We expect to recruit a minimum of 12 participants and to administer the program for 12 weeks at a frequency of two visits per week. We anticipate that implementing and supervising the home-based PA protocol will be feasible as determined by recruitment and retention rates, patients’ satisfaction, and compliance with the program. We also expect that this protocol will increase physical activity levels, improve general strength, balance, physical function, and reduce the risk of falls in sedentary older adults with T2DM. DISCUSSION/SIGNIFICANCE: As the third cause of death in PR, T2DM represents a public health challenge. An effective home-based PA program may decrease morbidity and mortality rates in older adults by increasing PA and functional health. This study will provide data for planning a randomized controlled trial to assess its effectiveness in the outcomes of interest.

Objectives/Goals: (1) Conduct a pilot study documenting prevalence of tinnitus in a sample of Puerto Rican adults at the Audiology Clinic of the Medical Sciences Campus-University of Puerto Rico, (2) categorize patterns of tinnitus, (3) document intervention received for tinnitus, and (4) study sociodemographic characteristics of Puerto Rican adult participants with tinnitus. Methods/Study Population: A descriptive retrospective study was performed reviewing 121 clinical records of patients seen at the Audiology Intramural Clinic of the Medical Sciences Campus of the Universidad de Puerto Rico between 2022 and 2023. They were analyzed to determine the prevalence of tinnitus among this cohort. The study was submitted to the Office of Human Participants for revision and approval under the exempt category. The data were used to categorize the type of tinnitus, episodic versus constant, tonal versus non-tonal and the sociodemographic description of the sample. Results/Anticipated Results: From these 121 records, 70.2% (n = 85) were females and 29% (n = 29.8) were males. Subject ages ranged between 21 and 65 years. About 30% reported being single 30.6% (n = 37), followed by 21.5% (n = 26) reporting being married. From the 62 revised clinical records of subjects that reported tinnitus, 24% (n = 29) classified their tinnitus, in terms of how long they experience its presence, as constant, while 14% (n = 17) classified their tinnitus as intermittent. From the 62 revised clinical records, 44 participants (36.4%) described their tinnitus as tonal and 64.6 % as a complex sound of those patients 38 (31.4%) reported the tinnitus as a high-frequency pitch sound. Of the 62 patient records, the majority (98.4%) informed that they never received the treatment for tinnitus. Discussion/Significance of Impact: The results indicate that more than half of adults evaluated in the UPR Audiology Intramural Clinic (51%) had tinnitus. Age range was broad developing at any age but most prevalent in middle-aged females. Manifested permanent as a tonal or a complex sound. About 98.4% informed that they never received treatment, therefore, there is a need to ensure intervention.