OBJECTIVES/GOALS: Atopic Dermatitis (AD) affects 10% of people globally and is studied widely in clinical trials. However, clinical outcomes assessment (COA) for AD are not standardized, hindering easy comparisons across different studies. This study examines AD studies to identify the most used COAs. METHODS/STUDY POPULATION: Clinicaltrials.gov was searched to identify AD trials conducted between 2011 and 2021. Ongoing and completed trials were classified according to the therapeutic modality: Biologic, Immunosuppressive, or Other (botanicals and antibiotics). Further, AD trials were examined to determine which of the COAs listed in the FDA compendium issued in 2021 were included: the Investigators Global Assessment (IGA), the Eczema Area and Severity Index (EASI-75), and the Pruritus Numeric Rating Scale (NRS). The results were analyzed to determine which COA is most frequently used and if there were differences across therapeutic modalities or trial phases. RESULTS/ANTICIPATED RESULTS: Across a total of 50 AD trials registered in clinicaltrials.gov, EASI-75 was the most used COA; the item was included in 12 of 16 biologics, 5 of 14 immunosuppressives, and 7 of 20 other products. Moreover, AD trials of biologics included more of the FDA-suggested COAs than those studying other modalities. There were also differences across the clinical trial phases in that most phase III AD trials (83.33%, n=8) included two of the three COAs listed (IGA and EASI-75) compared to less than half of phase I/II trials (vs. 43.75%, n=32). DISCUSSION/SIGNIFICANCE: Findings from this study indicate a lack of COA harmonization across AD trials, impeding comparative analysis of the trial results. Establishing a common standard for COA would foster communication and transparency among key stakeholders including researchers, healthcare providers, and patients.

OBJECTIVES/GOALS: The University of Michigan Frankel Cardiovascular Center (FCVC) Innovation Challenge is an annual competition offering funding for innovative ideas to improve cardiovascular care. Due to the COVID-19 pandemic, administrators converted the recruitment process and pitch event to fully virtual. METHODS/STUDY POPULATION: We detail the process of converting the event from a hybrid process (virtual and in-person recruiting and in-person event) to a fully virtual one. Changes to the event included implementing a virtual recruiting process utilizing short video recordings as submission format; a new tool for storing and displaying submissions; fully virtual finalist selection and coaching; and a fully virtual pitch and judging event. The submission process tracked information about submissions that include the type of idea (process or product), role of team lead, and department of team lead. RESULTS/ANTICIPATED RESULTS: The FCVC Innovation Challenge was successfully converted to a fully virtual event. Methods and tools will be shared to allow similar institutions to replicate a successful virtual pitch event. These include methods and tools utilized to allow participants to describe their ideas, strategies to select and coach finalists, and to host a virtual pitch event. Data will be shared on the number of ideas and category (product/process) of projects submitted, and number and category of finalists selected. DISCUSSION/SIGNIFICANCE: This case review can demonstrate how institutions can use a similar virtual idea submission and pitch process to (1) catalyze innovative ideas that can impact patient care by accessing its communitys ideas and (2) fund innovative ideas that do not fit traditional mechanisms.

OBJECTIVES/GOALS: This presentation reports activities of a NCATS-funded collaborative working group created to promote dissemination and implementation (D&I) research within the CTSA landscape. Our working group seeks to meet both the conceptual as well as practical challenges to advancing the utilization of D&I across the translational science spectrum. METHODS/STUDY POPULATION: A fundamental focus of D&I is supporting the movement of effective health interventions into real-world use so that they benefit population health. Yet, this process remains unpredictable, with some interventions receiving widespread uptake in practice and others (of similar potential benefit) failing to translate. The value of research efforts is wasted when directed toward the “wrong” interventions. Recent discussion and experience amongst investigators in our collaborative working group has resulted in new ways of addressing this problem. Specifically, tools borrowed from business and management have shown promise in predicting which health interventions have the highest potential for commercialization and dissemination. RESULTS/ANTICIPATED RESULTS: We will conduct an environmental scan of CTSA hubs to understand their approaches to supporting commercialization and business development around research products, identifying the most promising and effective methods and processes. We will compile various tools for identifying and supporting interventions with the highest potential for commercialization, including how to form the multidisciplinary and stakeholder-engaged teams necessary to make these determinations. Finally, we will further explore the differences between patentable and non-patentable innovations and make recommendations for CTSAs in supporting the latter. DISCUSSION/SIGNIFICANCE: Commercialization of non-patentable interventions is an essential and underexplored element of the translational science spectrum. The perspectives and methods of D&I should not be relegated to late-stage translational steps, but rather inform the conduct of translational science writ large.

OBJECTIVES/GOALS: The clinical management of differences of sex development (DSDs) aims to guarantee best practices in medical care while addressing concerns related to non-reversible surgeries. Rhetorical analysis was conducted to study the balance between science policy and patient advocacy related to DSD surgeries as depicted in medical education materials METHODS/STUDY POPULATION: Unrestricted transcripts of two educational videos and text from all chapters of a handbook addressed to medical learners and faculty by the Association of American Medical Colleges (AAMC) were submitted to automated word cloud analysis (NVivo, QSR International®). Words with a weighted percentage > 0.19% from total words of a given source were defined as words of frequent use and were selected for further analysis after exclusion of words as conjunctions, prepositions, pronouns, or conversational fillers. Words sharing noun, adjective and adverb forms were coded and weighed as a single word following the Oxford dictionary. Discrepancies on word selection, exclusion or coding were resolved between four raters. The rhetorical context of most frequent words was identified. RESULTS/ANTICIPATED RESULTS: The word cloud analysis of the video resource intended for medical learners (n=104 words of frequent use) and the video intended for medical faculty (n= 94 words of frequent use) depicts a patient-centered approach (word people’) that is based on expert opinion (word [I] think’). The handbook (n= 998 words of frequent use) makes reiterated reference to patients’; lgbt’; gender’; health’; and caring’ while underscoring health concerns that are unrelated to genital variance (health’; caring’ and medical’). The noun surgery’ did not figure among the most frequent words in spoken language nor in written text even when summing its adjective and adverb forms. DISCUSSION/SIGNIFICANCE: Educational materials by the AAMC on DSDs accentuate patient-centered care within a medical humanism framework. However, the lack of discussion of DSD surgeries is an educational gap that should be addressed by key science policy and patient advocacy stakeholders.

OBJECTIVES/GOALS: The present study sought to investigate kratom use motives among the U.S population with the goal of discovering previously unknown health-related reasons for kratom use by the general public. METHODS/STUDY POPULATION: To guide decisions regarding kratom regulation, the FDA solicited comments from the public regarding the abuse potential of these substances, medical usefulness, and impact of scheduling changes from July 2021 until August 2021. Comment participation was open to the public. The first 6,353 consecutive comments posted on the Federal Register website were retrieved and analyzed. Duplicate comments and comments not pertaining to kratom were excluded from the analysis. The comment submissions were reviewed and categorized using an inductive approach via thematic content analysis. RESULTS/ANTICIPATED RESULTS: Respondents reported over 108 independent health-related reasons for kratom self-medication. Most often fell under the categories of mental health (1911 counts), pain management (1873 counts), substance use disorder (1635 counts), rheumatic diseases (613 counts), and degenerative spine diseases (247 counts). Many comments (701 counts) reported use for miscellaneous purposes, which included to increase focus (212 counts), treat insomnia (127 counts), and decrease fatigue (99 counts). Neurological diseases (e.g., migraines, restless legs syndrome, and multiple sclerosis) and digestive disorders (e.g., irritable bowel syndrome and Crohns Disease) were also reported (147 and 96 counts, respectively). Respondents also reported on the abuse potential and adverse effects of Kratom (122 counts). DISCUSSION/SIGNIFICANCE: This is the first study to delineate and classify motives for kratom use among Americans. Individuals reported using kratom for a wide spectrum of health-related reasons. Though these results may be influenced by the placebo effect, they suggest that kratom alkaloids may possess therapeutic activity for previously unknown applications.

OBJECTIVES/GOALS: The present work aims to use baseline data to identify demographic, clinical, and psychosocial factors associated with patients who receive analgesic benefit from temporary neurostimulation. METHODS/STUDY POPULATION: This study presents baseline data from our descriptive, prospective, longitudinal study. Consecutive patients who present to the University of Arkansas for Medical Sciences Interventional Pain Management Clinic for implantation of a neurostimulation device, have met clinical criteria for implantation of a neuromodulation device, and are able to speak and understand English are invited to participate. Prior to the placement of the temporary stimulator, each patient completes demographic and symptom-related questionnaires. Clinical characteristics are obtained through medical record review. RESULTS/ANTICIPATED RESULTS: We anticipate enrolling 50 participants in order to have 30 patients that report analgesic benefit from temporary neurostimulation. Variability in demographics, clinical characteristics, and psychosocial factors will be reported between patients who receive and those who do not receive analgesia following temporary neurostimulation. Gender differences will also be reported. DISCUSSION/SIGNIFICANCE: Despite the use of varying outcome measures, studies to date have not incorporated validated patient reported outcomes or controlled for key demographic and clinical characteristics. Our analysis evaluates clinical and psychosocial variables associated with successful temporary neurostimulation.

OBJECTIVES/GOALS: Childhood survivors of ALL are at considerable risk for late effects which are exacerbated by excess weight. The proposed study involves the adaption and evaluation of the first empirically supported intervention for childhood survivors of ALL that is consistent with all national recommendations for the treatment of childhood obesity. METHODS/STUDY POPULATION: The proposed intervention will be adapted from family-based behavioral weight loss treatment (FBT) a multicomponent intervention which targets diet, activity, behavioral skills, parenting, and social facilitation among children and their parents. The Framework for Reporting Adaptations and Modifications-Enhanced structure (FRAME), a dissemination and implementation framework, will guide the adaptation, allowing for the incorporation of feedback previously gathered from key stakeholders. A single-arm, non-randomized trial of the adapted intervention will then be conducted to evaluate its acceptability, feasibility, and preliminary indications of efficacy including measures of relative weight change and associated health-related behaviors among 40 childhood ALL survivors and their families. RESULTS/ANTICIPATED RESULTS: Self-reported feedback from families at the end of treatment (EoT) is anticipated to demonstrate that this intervention will be regarded as both acceptable and feasible. Other measures of feasibility will include attendance and retention rates, which are expected to reflect to those of previous FBT trials (92% and 85%, respectively). Preliminary indications of the efficacy of the adapted intervention will be investigated through the comparison of a series of measurements taken at both baseline and EoT. Changes in relative weight will be assessed and are expected to meet a previously established range of clinically meaningful reduction in child percent overweight of 9 units or more. Improvements in dietary intake, physical activity, and health related quality of life are also anticipated. DISCUSSION/SIGNIFICANCE: Knowledge gained from the implementation of the first evidence-based intervention adapted for childhood survivors of ALL will be critical to the justification of a larger-scale, randomized controlled trial and holds promise to effectively modify the risk for chronic disease among a vulnerable population.

OBJECTIVES/GOALS: Minimal human investigations have assessed the effect of synbiotics (combination of pre- and probiotics) on anxiety symptoms, despite evidence from preclinical research. Our study aimed to determine the feasibility of a randomized, placebo-controlled trial utilizing synbiotics to reduce anxiety symptoms in older female breast cancer survivors. METHODS/STUDY POPULATION: We aimed to recruit older female breast cancer survivors experiencing anxiety symptoms to a 4-week randomized, placebo-controlled clinical trial. At commencement of the project, participants were eligible if they : 1) were 50-75 years old; 2) completed primary treatment for breast cancer; 3) were experiencing clinical anxiety symptoms 4) agreed to not change dietary supplements 5) were willing to comply with daily supplement regimen; and 6) were able to read and speak English. Use of anxiolytic or microbiome-altering medications, or changes to anxiety treatment within 4 weeks of enrollment were criteria for exclusion. Due to budgetary limitations, we were unable to recruit from state cancer registries, and instead recruited via newspaper advertisements and flyer distribution. RESULTS/ANTICIPATED RESULTS: One participant has successfully been recruited and completed the duration of the clinical trial, and two others have expressed interest but were deemed ineligible. Barriers in recruitment resulted in the following modifications to protocol: we expanded our eligibility criteria by removing the upper age limit (now 50+ years old) and now are recruiting females with a personal or family history of breast cancer. We partnered with the Spencer Cancer Center of East Alabama Health to aid in recruitment. DISCUSSION/SIGNIFICANCE: Integrative approaches to improved patient outcomes are needed, however, recruitment remains a paramount barrier for clinical trials. Addressing our issues for recruitment has opened eligibility to more individuals and allows us to continue our investigations, answer our research questions, and advance translational science.

OBJECTIVES/GOALS: Vascular cognitive impairment (VCI) is the leading cause of dementia behind Alzheimers Disease (AD) and is often the result of brain hypoxia. Diets rich in polyunsaturated fatty acids (PUFAs) can lower cognitive decline and AD incidence in human patients. Therefore, our goal is to determine the mechanisms that PUFAs influence in a mouse model of VCI. METHODS/STUDY POPULATION: We hypothesize that hypoxia promotes endothelial P-tau accumulation and vasotrophic uncoupling, impairing endothelial integrity. Additionally, we believe that a preventative PUFA-enriched diet blocks this uncoupling and subsequently prevents/delays neurovascular dysfunction and cognitive decline. Male and female mice will be administered a control or novel PUFA-enriched dietary intervention 1 month prior to hypoxic injury using the bilateral carotid artery stenosis model. Mice will continue their diet and be assessed for cerebral blood flow, cognitive function, and motor function at 1- & 3-month time points. Following euthanasia, tissue samples from deep cortical regions and microvasculature will be examined for endothelial- & neuronal-specific P-tau accumulation, inflammation, and cell death. RESULTS/ANTICIPATED RESULTS: Preliminary data in our lab indicates that hypoxia leads to a two-fold increase in endothelial P-tau accumulation and lowered mature BDNF (mBDNF) in brain microvascular endothelial cells (BMECs) compared to controls. Further, BMECs cultured in media with the PUFA docasahexaenoic acid (DHA) had lowered P-tau and increased mBDNF after hypoxia compared to controls. Based on this data and past research, we anticipate that mice on the PUFA-enriched diet will have enhanced cognitive and motor function alongside improved cerebral blood flow compared to controls. Also, we expect that mice on our PUFA-enriched diet will have lowered tau pathology, cell death, and neuroinflammation alongside increased blood brain barrier integrity and altered fatty acid composition in brain and vascular tissue samples. DISCUSSION/SIGNIFICANCE: An AHA Presidential Advisory identified cognitive function as modifiable through the management of cardiovascular risk factors, like diet. However, the mechanisms underlying the benefits of PUFA-enriched diets are unknown. Successful completion of these studies will provide insight into the vaso-neuronal protective effects of PUFAs in VCI.

OBJECTIVES/GOALS: The SARS-CoV-2 (Severe Acute Respiratory Syndrome CoronaVirus-2), which underlies the current COVID-19 pandemic, among other tissues, also targets the central nervous system (CNS). The goal of this study is to investigate mechanisms of neuroinflammation in Lipopolysaccharides (LPS)-treated mouse model and SARS-CoV-2-infected hamsters. METHODS/STUDY POPULATION: In this research I will assay vascular reactivity of cerebral vessels to assess vascular dysfunction within the microcirculation. I will determine expression of proinflammatory cytokines, coagulation factors and AT1 receptors (AT1R) in isolated microvessels from the circle of Willis to assess inflammation, thrombosis and RAS activity in the microvasculature. LPS and SARS-CoV-2, are both associated with coagulopathies and because of that I will measure concentration of PAI-1, von Willebrand Factor, thrombin and D-dimer to assess the thrombotic pathway in the circulation. Histology and immunohistochemistry will assess immune cell type infiltration into the brain parenchyma, microglia activation and severity of neuroinflammation and neural injury. RESULTS/ANTICIPATED RESULTS: We hypothesize that under conditions of reduced ACE2 (e.g., SARS-CoV-2 infection), AT1R activity is upregulated in the microvasculature. In the presence of an inflammatory insult, these AT1Rs promote endothelialitis and immunothrombosis through pro-thrombotic pathways and pro-inflammatory cytokine production leading to endothelial dysfunction in the microvasculature, blood brain barrier (BBB) injury, deficits in cognition and increased anxiety. We will test this hypothesis through 2 aims: Aim 1: Determine the role of the pro-injury arm of the RAS in the pathophysiology of the brain in animal models of neuroinflammation and COVID-19. Aim 1: Determine the role of the protective arm of the RAS in the pathophysiology of the brain in animal models of neuroinflammation and COVID-19. DISCUSSION/SIGNIFICANCE: This study will provide insights that will complement on-going clinical trials on angiotensin type 1 receptor (AT1R) blockers (ARBs) in COVID-19. This research is a necessary first step in understanding mechanisms of brain pathogenesis that can set the groundwork for future studies of more complex models of disease.

OBJECTIVES/GOALS: The complex health profile of those supervised by community corrections places them at a greater risk of morbidity and mortality from social disruption, infection, and of substance misuse, relapse, and overdose. This study aims to explore individual and social determinants of SUD and treatment utilization for this population. METHODS/STUDY POPULATION: A secondary data analysis was conducted using an administrative database from the Department of Corrections of Puerto Rico (DoC-PR) that included individuals under community supervision between 2015 and 2018 (N=13,163). Two logistic regression analysis were performed to predict SUD and SUD treatment utilization. The predictors included in the models were: a) age, b) gender, c) civil status, d) education level, e) employment status, f) income, g) number of dependents, h) housing zone, I) type of crime, j) offender classification k) community sentence classification, and k) sentence duration. RESULTS/ANTICIPATED RESULTS: A total of 4,531 cases (34.4%) were identified with an SUD and of these 79.2% were enrolled in treatment. Significant predictors of SUD included a parole sentence (vs probation), commission of felony, decreasing sentence duration and recidivism. Significant demographic variables include, male, single, younger age, unemployed, residing in an urban zone and decreasing income. Significant predictors for SUD treatment utilization were a probation sentence, older age and residing in a rural zone. Mandated treatment may explain a higher likelihood of treatment utilization, yet prevalent modalities consist of residential abstinence based, non-professionalized services known to have poor treatment outcomes. The current data set does not include follow up data to assess changes in treatment utilization. DISCUSSION/SIGNIFICANCE: We should aim to prevent health and social disparities and risk of sentence revocation associated with interventions that lack evidence to support their effectiveness. Next steps should address challenges and opportunities for the adoption of EBPPs for individuals with an SUD under community corrections supervision.

OBJECTIVES/GOALS: To evaluate whether a low cost, functional dynamic 3-D printed upper extremity (UE) orthosis could be designed, fabricated and used by children with cerebral palsy (CP) with severe unilateral UE involvement and the ability of common standardized instruments to assess its effectiveness. METHODS/STUDY POPULATION: Five patients, ages 13-17 years, with CP and unilateral UE involvement, MACS levels III-IV, were enrolled. Custom forearm thumb opponens orthosis and dynamic upper extremity orthosis (DUEO) were designed and fabricated by a multidisciplinary team for use during 8 one-hour occupational therapy sessions targeting bimanual UE training. Pre- and post-assessments included Assisting Hand Assessment(AHA), Melbourne-2(MA-2), Pediatric Motor Activity Log-Revised(PMAL-R), and PedsQL Measurement Model for the Pediatric Quality of Life Inventory: CP Module(Peds-QL). RESULTS/ANTICIPATED RESULTS: The 3D printed orthotic device is custom fit to the patient based on scans of their arm and is designed with a tensioning system that allows for functional usage of the arm. It incorporates a rigid polymer to provide support and flexible material for comfort where appropriate. Overall, higher post-treatment scores were found for the majority of participants using the custom orthotic. Four made minimal clinically important differences (MCID) in the AHA. Three met MCID scores in subtests of MA-2 (two positive and one negative). Patient-reported outcome improvements were found for PMAL-R for four, but only one met MCID and at least three reported improvements in more than one domain of PedsQL. DISCUSSION/SIGNIFICANCE: Children with CP may often present with UE impairment, yet current therapeutic orthoses only target the progression of contractures and are still limited by cost and discomfort. Our team designed and fabricated a functional, low cost, 3D printed orthosis that showed significant gains in UE function.

OBJECTIVES/GOALS: Perioperative surgical care is team-based with close partnership between surgeons, residents, advanced practice professionals (APPs), and others. The objective is to develop an understanding of the current state and implementation needs required for APPs to engage surgical patients in advanced care planning (ACP) to promote goal concordant care. METHODS/STUDY POPULATION: We will conduct a mixed methods evaluation of ACP knowledge, attitudes, and beliefs amongst surgical APPs to identify barriers and facilitators of APPs engaging in a team-based approach to engaging surgical patients in ACP. We will conduct an online survey and qualitative interviews in the following 4 domains: 1) knowledge, skills, and attitudes about engaging in ACP with a patient or their surrogate decision maker during their perioperative care; 2) prior ACP-specific education; 3) experiences conducting ACP discussions with patients; and 4) perceived training needs to increase ACP uptake and documentation. The findings will provide the foundations to design team-based interventions focused on addressing the barriers and inform training and coaching needs to develop expertise and comfort in the ACP process. RESULTS/ANTICIPATED RESULTS: We expect variability in the knowledge, skills, attitudes, and experiences with the ACP process. We anticipate gaining a better understanding of the educational materials best suited to support APPs as they begin engaging patients in ACP. Possible barriers to APP-led ACP discussions include inconsistent role delineation, uncertainty about the value of pre-operative vs. post-operative ACP discussions, lack of experience engaging in ACP discussion, and lack of familiarity with electronic health records ACP tools. Possible facilitators of APP-led ACP discussions may be related to past work experience settings, exposure to ACP in educational preparation, hands-on observation of value of ACP in surgical patients and influences from attending and residents. DISCUSSION/SIGNIFICANCE: While current ACP research in surgery focuses on physician-led patient engagement in ACP discussions, there is a paucity of research focusing on how to develop a team-based approach to ACP discussions in surgery. This study will provide information necessary for the development of interventions that increase team-based ACP for surgical patients.

OBJECTIVES/GOALS: Fragile X-associated tremor/ataxia syndrome (FXTAS) is a devastating rare neurological disorder that negatively impacts movement and cognition. To date, there are no effective pharmacological treatments for FXTAS. Our goal was to develop a cell culture model of FXTAS to investigate promising therapeutics. METHODS/STUDY POPULATION: To establish mitochondrial dysfunction, normal human cell lines and human-induced pluripotent cells were treated with multiple concentrations of glucose/ glucose oxidase (GluOx) at 2,12, and 24 hour time points to induce varying intensities of oxidative stress. The degrees of oxidative stress were measured by apoptosis and mitochondrial reactive oxygen species (ROS) production. Curcumin and MSKE compounds effective against oxidative damage in mitochondria were used to rescue glucose oxidase-induced oxidative damage in both cell lines. To test the ability of these drugs to restore mitochondrial health, cell viability and cellular superoxide production were assessed by propidium iodide and the MitoSox fluorescence assay, respectively. RESULTS/ANTICIPATED RESULTS: We anticipated that GluOx at varying concentrations and time points would proportionally increase levels of apoptosis and mitochondrial ROS, reflective of mitochondrial damage, with the most severe dysfunction occurring at a dose of 25 nM and the longest duration of 24-hr exposure. Administration of MSKE in concentrations ranging from 10-8 to 10-5 M in half log increments, did not reverse the oxidative defects induced in the cell lines. However, curcumin concentrations increased cell viability at the 2, 12, and 24 hour time period. Results indicate that the research design should be modified by increasing the concentration of both glucose and MSKE to provide a reliable test of the hypothesis. DISCUSSION/SIGNIFICANCE: These studies illustrate the usefulness of this in vitro model to test novel therapeutics in neuronal FXTAS models and expand the discovery of mitochondrial markers for the syndrome.

OBJECTIVES/GOALS: We aim to determine whether non-neuronal, non-synaptic glutamate signaling mechanisms can be targeted to produce highly specific, narrow changes in brain function that would benefit CNS disorders. To do this, we investigated cognitive changes produced through manipulating the activity of the astrocytic glutamate release mechanism system xc-. METHODS/STUDY POPULATION: System xc- (Sxc) activity was eliminated by mutating the gene Slc7a11 through pronuclear injection of zinc-finger nucleases into Sprague Dawley rat embryos to create a line of rats lacking Sxc (MSxc rats). To confirm a lack of Sxc activity, we verified that tissue from MSxc rats had a complete lack of xCT, which is the regulatory subunit of Sxc that is encoded by Slc7a11. We also verified that astrocyte cultures generated from MSxc tissue lacked cystine-evoked glutamate release. Next, we measured development (body weight), CNS regulation of metabolism, and other indicators of generalized, non-specific brain function as well as behaviors that are reliant on executive function, such as cognitive flexibility, impulse control, decision-making, and response inhibition. RESULTS/ANTICIPATED RESULTS: Eliminating Sxc was not lethal and did not impair development or produce widespread changes in brain function as is commonly observed when deleting other glutamate mechanisms. MSxc rats did not differ from wildtype in growth rate, central regulation of metabolism as reflected by absolute or diurnal changes in core body temperature, locomotor activity in a familiar or novel environment, or simple forms of cognition such as novel object recognition, or operant responding (food and cocaine-reinforced). In contrast, behaviors that rely on executive function were impaired. MSxc rats displayed deficits in cocaine reinstatement and attentional set-shifting. We anticipate MSxc rats to also show impairments in decision-making in the rat gambling task and response inhibition in the stop-signal reaction time task. DISCUSSION/SIGNIFICANCE: Eliminating Sxc activity in rats produced deficits in behaviors reliant on executive function without impacting development or simple brain function. These results highlight the potential of targeting Sxc to enhance cognition without generating therapeutically limiting adverse effects resulting from non-specific changes in brain function.

OBJECTIVES/GOALS: Our hypothesis is that microneedle array (MA) extraction of interstitial fluid (ISF) will enable minimally invasive quantitation of heavy metal (HM) exposure. We aim to establish analytical parameters for ICP-MS analysis of HMs, quantify baseline HM content in ISF vs other fluids, and characterize a mixed HM exposure model. METHODS/STUDY POPULATION: Ten healthy human volunteers were recruited into the study, approved by the UNM Human Research and Resource Committee Each subject had blood and urine collected. ISF was also collected using 3D-printed MAs inserted into the forearm. Additionally, twelve Sprague Dawley rats were unexposed (n=6) or exposed (n=6) to ad libitum water containing a mixture of uranium (U), cadmium (Cd), vanadium (V), and arsenic (As), each at 5X the maximum contaminant level (MCL) for drinking water under a protocol approved by the UNM animal care and use program. Human and animal fluids were analyzed, using ICP-MS, to quantify the levels of U, Cd, V, and As. RESULTS/ANTICIPATED RESULTS: Recent advances in ISF extraction and analysis suggest a minimally invasive method that can be adapted to monitor HM exposure and biological loads longitudinally in both localized and dispersed communities. ISF can be collected with MAs and is a rich source of disease and exposure biomarkers. However, determining biological loads remains challenging due to the need to collect blood or urine from a dispersed rural population over time. Our preliminary results suggest similar HM concentrations in ISF, compared with blood in small unexposed human and animal populations. All four metals can be successfully quantified in tandem using ICP-MS. DISCUSSION/SIGNIFICANCE: Chronic exposure to heavy metals (HM) is associated with detrimental health effects. Exposure to multiple HMs is suspected to have additive or synergistic harmful effects. We envision a wearable microneedle patch that could be mailed to individuals or distributed through community centers, worn for a few hours, and returned to a central laboratory.

OBJECTIVES/GOALS: Supervision is a promising strategy for supporting high-fidelity implementation of evidence-based practices (EBPs) in substance use disorder (SUD) settings. In this study, we explore current supervision practices in community SUD programs and identify organizational and environmental factors that shape them. METHODS/STUDY POPULATION: We interviewed 25 leaders and counselors at 8 community SUD programs in Arkansas, and 16 leaders at external stakeholder organizations (e.g., regulators, payers, licensing boards). Interview guides were based on the i-PARIHS framework. Interviews were conducted on Zoom or phone, lasted ~1 hour, and were recorded and transcribed. Below we outline findings based on preliminary analyses; full thematic analyses will be completed before presentation. RESULTS/ANTICIPATED RESULTS: Participants generally recognized the importance of utilizing EBPs and supporting their use through supervision. Counselors professional backgrounds and training vary substantially, necessitating continuing education and supervision. However, different professional, licensing and regulatory standards create a complex web of requirements and practices. Supervisors typically require clinical experience, but are rarely trained in supervision. They are internal or external to the organization, and provide individual or group supervision. Supervisors most often rely on case summaries and chart reviews, sometimes on direct observations and role-playing, and rarely (one program) on session recordings. Supervision goals are broad, and while EBP use is encouraged, it is rarely the focus of supervision. DISCUSSION/SIGNIFICANCE: To enhance supervision in community SUD settings and improve implementation of EBPs, new supervision strategies need to fit the various norms, expectations, and standards (e.g., professional, regulatory) that characterize community SUD programs, their workforce, and their environments.

OBJECTIVES/GOALS: Studies using Medicare data indicate that physical therapists (PTs) and physical therapist assistants (PTAs) are not providing falls prevention to at-risk older adults in rehabilitation. We aimed to identify PTs and PTAs knowledge and use of the Centers for Disease Control and Prevention’s STEADI fall prevention toolkit. METHODS/STUDY POPULATION: We conducted a cross-sectional survey distributed to a convenience sample of PTs and PTAs in the United States through email blasts and social media. Descriptive statistics were used to summarize the demographic characteristics of the respondents. Some categorical variables were combined to provide more meaningful classifications or due to small frequencies. We used independent samples t-tests for continuous data, and chi-square and Fisher’s Exact tests for categorical data to compare characteristics between respondents that do and do not conduct falls risk screenings.Frequency counts and percentages were used to summarize survey responses related to falls risk screening and knowledge/use of STEADI. SAS® version 9.4 was used for all statistical analyses. RESULTS/ANTICIPATED RESULTS: PTs and PTAs (N = 425) who responded to the survey and worked in clinical settings with older adults were included. Eighty-nine percent of respondents reported conducting clinical falls risk screening, yet only 51% were 'familiar’ to 'very familiar’ with STEADI. Twenty-two percent of respondents were not familiar at all with STEADI. Of the respondents who were 'very familiar with the STEADI (n = 132, 31.1%), 84.1% (n = 111) reported using STEADI in clinical practice. Seventy-six percent of respondents who use the STEADI implemented it by choice even though the majority (52.1%, n = 63) did not have it embedded in their workflow or documentation. DISCUSSION/SIGNIFICANCE: PTs and PTAs in the United States have some familiarity with and use STEADI in clinical falls prevention, and those who are very familiar with it use it by choice. Further research is needed to address the knowledge gap of STEADI and support PTs and PTAs providing falls prevention to older adults attending rehabilitation.

OBJECTIVES/GOALS: To identify the role of sialoglycans in the mechanisms underlying cancer-associated fibroblast-derived exosomes (CAFEX) immuno-regulatory properties. The central hypothesis is that CAFEX manipulates the immune response to allow immunoevasion and glycomic approaches can identify the signaling factors involved. METHODS/STUDY POPULATION: Cancer associated fibroblasts (CAFs) were isolated from primary head and neck tumors, expanded, characterized and cryopreserved prior to experimentation and isolation of CAFEX. Sialoglycan expression was determined using lectin-specific staining of cells and bead-captured CAFEX in combination with flow cytometry analysis. Siglec expression and expression of M2-macrophage markers were also determined by flow cytometry analysis and cytokine bead arrays. Inhibition studies involved either the enzymatic removal of cell-surface sialoglycans or alternatively, a specific small molecule analog inhibitor of sialoglycan transferases. RESULTS/ANTICIPATED RESULTS: Both CAFs and CAFEX expressed abundant levels of cell-surface sialoglycans. CAFEX induced an M2-macrophage phenotype in primary monocytes, based on surface marker expression and cytokine secretion profiles. The induction of the M2 phenotype in macrophages was attenuated upon the removal of sialoglycans from the surface of CAFEX either by enzymatic removal or via a small molecule inhibitor. CAFEX were also able to directly bind members of the Siglec family, which are sialoglycan specific lectin receptors expressed on immune cells, including macrophages. DISCUSSION/SIGNIFICANCE: Collectively, these studies suggest that surface presentation of sialoglycans by CAFEX may induce an immunosuppressive phenotype in monocytes/macrophages. Consequently, this may be a novel mechanism by which cells within the tumor bed facilitate immunoevasion during tumor progression.

OBJECTIVES/GOALS: Inflammation-promoting exposures, including hyperglycemia and inflammatory cytokines, are common in the development and progression of multiple cancers. Here, we determine if high glucose drives the dysregulation of XRCC1, changing BER/SSBR capacity, and increasing resistance to DNA damaging agents. METHODS/STUDY POPULATION: Here, we acutely exposed cell lines models with 30 mM glucose to mimic hyperglycemic changes and monitored the gene and protein expression of STAT3 and XRCC1. We selected the osteosarcoma cell line U2OS, with high STAT3 activation before glucose challenge, and the non-tumorigenic human embryonic kidney cell line HEK293T, with low STAT3 activation before glucose challenge, to dissect the role STAT3 plays in dysregulating DNA repair. We also examined changes in STAT3 occupancy at the XRCC1 promoter following glucose challenge using chromatin immunoprecipitation (ChIP). Finally, we measured changes in the sensitivity to the alkylating agent methyl methanesulfonate (MMS) induced by the glucose challenge using cell survival and DNA strand break analysis. RESULTS/ANTICIPATED RESULTS: High glucose challenge increases the phosphorylation and activation of STAT3 and subsequently increases XRCC1 gene and protein content in the cell. Acute high glucose activated STAT3, driving the subsequent expression of XRCC1 in U2OS and HEK293T cells through increased STAT3 occupancy at the XRCC1 promoter. High glucose also reduced sensitivity to MMS, increasing cell survival. The most significant increase in resistance to MMS occurred in the HEK293T, which also showed the largest increase in XRCC1 protein expression following the glucose challenge. Increased survival correlated with the faster resolution of DNA strand breaks in glucose-challenged cell lines. DISCUSSION/SIGNIFICANCE: This work has identified a novel regulatory mechanism by which high glucose drives the expression of XRCC1 through STAT3 activation, increasing DNA repair and resistance to the DNA damaging agent MMS. These data suggest dietary choices induce sustained XRCC1 expression and may contribute to chemoresistance and poor survival outcomes in cancers.