OBJECTIVES/GOALS: The goals of this project are to: i) investigate the cargo such as immune mediators (cytokines) and small non coding RNAs (sncRNAs) of EVs derived from nasopharyngeal secretions (NPS) of children with episodes of viral infections and exposed to SHTS; ii) test the biological activity of EVs released from upper airway mucosa on target/recipient lung cells METHODS/STUDY POPULATION: EVs were isolated from ten NPS samples of children with episodes of acute respiratory infections. EVs were characterized by particle sizing (size and concentration), EV markers, and protein arrays for interferons, cytokines, and other immune mediators content. RNA was extracted from ten samples of NPS-derived EVs by column for next generation high throughput sequencing (NGS) to identify sncRNAs in EVs. In studies currently in progress, EVs will be isolated from RSV-infected human nose organoids (HNOs) cells in air liquid interface (ALI) culture, with or without pre-exposure to tobacco smoke. EVs will be then tested for antiviral activity on recipient RSV-infected lower airway cells. Viral titers will be measured in recipient infected lung cells. RESULTS/ANTICIPATED RESULTS: We isolated EVs from NPS samples and confirmed by immunoblot EV markers CD63 and Alix. The average size of NPS-derived EVs of virus positive and negative patients was 170 nm and 145 nm, respectively. We determined the particles number of EVs, concentrations of IFN-βand IFN-λin NPS and NPS-derived EVs of these children. While IFN-βlevels were below the limit of detection in both NPS and NPS-derived EVs of all children, IFN-λwas detected in NPS and NPS-derived EVs from infected patients, except for the two patients with no viral infections. We extracted RNA from control-, virus infected- and/or SHTS- EVs and found that piR-36511, piR-40926, piR-49645, piR-32679 and piR- 53263 were all significantly enriched in EVs derived from NPS of children exposed to TS compared to those not exposed. DISCUSSION/SIGNIFICANCE: RSV leads to approximately 22,000 hospitalizations of children due to second-hand smoke. A vaccine is not currently available for RSV infection. EVs represent a novel translational approach to target undruggable. Airway mucosa EVs in viral respiratory infection function as antiviral messengers and tobacco smoke impairs the EV antiviral activity.

OBJECTIVES/GOALS: A particularly debilitating consequence of stroke is alexia, an acquired impairment in reading. Cognitive models aim to characterize how information is processed based on behavioral data. If we can concurrently characterize how neural networks process that information, we can enhance the models to reflect the neuronal interactions that drive them. METHODS/STUDY POPULATION: There will be 10 unimpaired adult readers. Two functional localizer tasks, deigned to consistently activate robust language areas, identify the regions of interest that process the cognitive reading functions (orthography, phonology, semantics). Another task, designed for this experiment, analyses the reading-related functional-connectivity between these areas by presenting words classified along the attributes of frequency, concreteness, and regularity, which utilize specific cognitive routes, and a visual control. Connectivity is analyzed during word reading overall vs. a control condition to determine overall reading-related connectivity, and while reading words that have high vs. low attribute values, to determine if cognitive processing routes bias the neural reading network connectivity. RESULTS/ANTICIPATED RESULTS: The localizer analysis is expected to result in the activation of canonical reading areas. The degree of functional connectivity observed between these regions is expected to depend on the degree to which each cognitive route is utilized to read a given word. After orthographic, phonologic, and semantic areas have been identified, the connectivity analysis should show that there is high correlation between all three types of areas during reading compared to the control condition. Then the frequency, regularity, and concreteness of the words being read should alter the reliance on the pathways between these area types. This would support the hypothesized pattern of connectivity as predicted by the cognitive reading routes. Otherwise, it will show how the neural reading network differs from the cognitive model. DISCUSSION/SIGNIFICANCE OF IMPACT: The results will determine the relationship between the cognitive reading model and the neural reading network. Cognitive models show what processes occur in the brain, but neural networks show how these processes occur. By relating these components, we obtain a more complete view of reading in the brain, which can inform future alexia treatments.

ABSTRACT IMPACT: Implementation of proteomics as a diagnostic tool for Nontuberculous mycobacteria (NTM) infection can provide a more accurate, efficient and cost-effective means for effectively diagnosing disease and enacting timely management decisions which can revolutionize patient care. OBJECTIVES/GOALS: Proteomic analysis is a proven diagnostic modality enabling rapid identification of microorganisms. We sought to apply proteomics to detect proteins unique to the most clinically relevant NTM. We then determined whether these unique proteomes could be used to successfully identify NTM species from in vitro cocktail preparations. METHODS/STUDY POPULATION: NTM reference strains for M. avium, m. intracellulare, m.chimaera, m. abscessus abscessus, m. abscessus massiliense and m. abscessus boletti were cultured in vitro and subjected to proteomic analysis using Liquid Chromatography tandem-Mass Spectrometry (LCMS). Tandem Mass Tag (TMT) data acquisition utilized an MS3 approach for data collection using Proteome Discoverer 2.4.

A comparative analysis of the proteome of each of these six species was performed quantitatively using LCMS. The process was repeated for three technical replicates and analyzed using the SEQUEST algorithm. Only high scoring peptides were considered utilizing a false discovery rate (FDR) of 1%. Once species-specific proteins were identified, we validated detection in individual and mixed samples of the six reference strains. RESULTS/ANTICIPATED RESULTS: The proteomic profiling of the six NTM reference strains successfully demonstrated proteins unique to each of the MAC species and MABC subspecies. Proteomic MAC species analysis produced between 327 to 2,540 unique peptides for each of the 3 species. MABC proteomic analysis identified between 17-74 unique peptides for each of the 3 subspecies. Fifteen different mixed preparations of MAC and MABC were then subjected to LCMS analysis and compared against the proteome profiles already curated for the six strains. We accurately identified at least one NTM in the majority of the samples (10/15). In three samples (3/15), the NTM was not correctly identified; in two of the samples (2/15) we were unable to determine the identity of NTM within the preparation. Further database curation will be performed to hone these results. DISCUSSION/SIGNIFICANCE OF FINDINGS: Proteomic analysis of in vitro reference strains successfully demonstrated protein fingerprints specific to six common disease-causing strains of NTM. Such findings can be used to evaluate clinical samples enabling more efficient diagnostic specificity. Further research will focus on identification of NTM in sputum samples of infected patients.

OBJECTIVES/SPECIFIC AIMS: The objective of this study was to prospectively assess caregiver-perceived barriers to accessing post-acute care for their injured child and determine if caregivers report ongoing, unmet health needs for their children after trauma. METHODS/STUDY POPULATION: This was a prospective cohort study that followed 50 participants for 6 months and administered surveys to parents of children who are admitted to a pediatric level 1 trauma center for injury. Surveys were given bi-weekly regarding care children received after hospital discharge. At 3 months, parents were surveyed over the phone on whether they were able to access all needed health services and if there were any perceived barriers to obtaining or providing at-home care. At 6 months, parents were given the Child & Family Follow-up Survey to assess ongoing physical, mental, social, and scholastic needs. Free responses and transcribed interviews were analyzed using thematic content analysis and frequencies are reported for discrete data. RESULTS/ANTICIPATED RESULTS: Out of 50 families recruited, 47 completed follow-up assessments. At 3 months, common themes regarding challenges after hospital discharge included difficulty scheduling specialist care; uncertainty in managing their child’s pain; transitioning home without enough knowledge to meet their child’s medical needs; lack of communication between multiple providers; distress at having providers release children to full activities before caregivers were comfortable. At 6 months, approximately 24% of parents reported children had ongoing cognitive limitations, 29% reported emotional problems, 19% reported physical limitations, 33.3% reported difficulty in school, and 15% reported play/social difficulties. DISCUSSION/SIGNIFICANCE OF IMPACT: Evidence suggests families face significant barriers in accessing follow-up care, despite nearly universal health insurance coverage for children. Further, a large percentage of parents report ongoing health needs, despite the majority of the cohort having only mild or moderate severity injuries. Making follow-up care more patient-centered for families of traumatically injured children may improve compliance with medical regiments and reduce the likelihood of future disability. Examples of this may be coordinating care among multiple specialty providers, so that patients with multiple injuries can schedule multiple follow-up appointments on the same day. Additionally, more caregiver education on administering pain medication, caring for wounds, and safe practices for returning to full activities would be beneficial for families.

OBJECTIVES/GOALS: IL-12 has potent immune effects but the presence of myeloid-derived suppressor cells (MDSC) can inhibit IL-12-induced NK cell cytotoxicity. Thus, we hypothesized that combining IL-12 with trabectedin, an immunosuppressive myeloid cell depleting agent, would improve its therapeutic efficacy in triple negative breast cancer (TNBC). METHODS/STUDY POPULATION: Combination IL-12 and trabectedin was tested in the 4T1 mouse model of TNBC. 4T1 cells were injected into the mammary fat pad of female BALB/cj mice. When tumors reached 50 mm3, mice were randomly divided into 4 groups and treated with PBS, IL-12 (0.5 μg/mouse 3x/wk), 0.15 mg/kg trabectedin weekly or the combination. Tumor volumes were measured by calipers. Mass cytometry was performed on spleens and tumors using a 35-antibody panel. Plasma IFN-γ levels were measured by ELISA. The role of NK cells was evaluated via depletion with anti-asialo-GM1. The Luminex Discovery Assay was used to measure plasma cytokines and immunohistochemistry was performed for CD4 and CD8a. Linear/nonlinear mixed effects modeling was used for in vivo data analysis and applicable t- or ANOVA tests were used for in vitrodata analysis. RESULTS/ANTICIPATED RESULTS: Combination IL-12 and trabectedin led to a significant reduction in tumor burden compared to single-agent IL-12, trabectedin and control treatments (all p<0.001), as well as higher levels of IFN-γ (all p<0.04). One combination treated mouse had complete tumor regression. Splenic MDSC were significantly decreased in combination treated mice. NK depletion abrogated the effects of combination therapy. Compared to mice receiving a control antibody, NK depletion of combination treated mice led to lower levels of CCL5 (p<0.01) and CXCL10 (p<0.001) and significantly higher tumor burden (p=0.001). CD8+T cell levels were significantly higher in combination treated mice compared to those receiving IL-12 (p<0.01), and these levels were decreased when mice were depleted of NK cells (p=0.01). DISCUSSION/SIGNIFICANCE: TNBC represents 15% of all breast cancer diagnoses and is associated with a worse prognosis compared to other subtypes. Black women are twice as likely to be diagnosed with TNBC and more likely to die from disease than White women. Thus, there is an increasing need to develop additional therapeutic options for this disease.

Project management expertise is employed across many professional sectors, including clinical research organizations, to ensure that efforts undertaken by the organization are completed on time and according to specifications and are capable of achieving the needed impact. Increasingly, project leaders (PLs) who possess this expertise are being employed in academic settings to support clinical and preclinical translational research team science. Duke University’s clinical and translational science enterprise has been an early adopter of project management to support clinical and preclinical programs. We review the history and evolution of project management and the PL role at Duke, examine case studies that illustrate their growing value to our academic research environment, and address challenges and solutions to employing project management in academia. Furthermore, we describe the critical role project leadership plays in accelerating and increasing the success of translational team science and team approaches frequently required for systems biology and “big data” scientific studies. Finally, we discuss perspectives from Duke project leadership professionals regarding the training needs and requirements for PLs working in academic clinical and translational science research settings.

Neonatal sepsis is a major cause of death and disability in newborns. Commonly used biomarkers for diagnosis and evaluation of treatment response lack sufficient sensitivity or specificity. Additionally, new targets to treat the dysregulated immune response are needed, as are methods to effectively screen drugs for these targets. Available research methods have hitherto not yielded the breakthroughs required to significantly improve disease outcomes, we therefore describe the potential of zebrafish (Danio rerio) larvae as preclinical model for neonatal sepsis. In biomedical research, zebrafish larvae combine the complexity of a whole organism with the convenience and high-throughput potential of in vitro methods. This paper illustrates that zebrafish exhibit an immune system that is remarkably similar to humans, both in terms of types of immune cells and signaling pathways. Moreover, the developmental state of the larval immune system is highly similar to human neonates. We provide examples of zebrafish larvae being used to study infections with pathogens commonly causing neonatal sepsis and discuss known limitations. We believe this species could expedite research into immune regulation during neonatal sepsis and may hold keys for the discovery of new biomarkers and novel treatment targets as well as for screening of targeted drug therapies.

There is increasing recognition of the crucial need for robust community engagement in health research and clinical trials. Despite this awareness, challenges persist in bridging the gap between researchers and communities. Much of the current discourse focuses on addressing issues such as cultural humility and equitable partnerships. To expand this conversation, we conducted community engagement studios, following the model by Joosten et al. We wanted to gather perspectives on research involvement across New Mexico. This process and resultant findings offer valuable insights into effective community engagement practices and advance clinical and translational science by amplifying community voices and needs.

OBJECTIVES/SPECIFIC AIMS: The aim of this study is to understand how porosity and collagen filling impact cell proliferation and differentiation in 3D printed scaffolds. METHODS/STUDY POPULATION: 3 groups of scaffolds will be 3D printed using FDM: solid scaffold, porous scaffold and porous scaffold with collagen gel (n=10 for each group) Internal geometries and surface structure will be analyzed using micro CT and Scanning Electron Mi RESULTS/ANTICIPATED RESULTS: We hypothesize that porosity and collagen filler will increase signal from Picogreen assay and ALP assay when normalized to scaffold surface area, indicating enhanced cell proliferation and differentiation. DISCUSSION/SIGNIFICANCE OF IMPACT: 3D printing PCU is a relatively new technique with very little published in the literature. Previous work has focused on the mechanical properties and not the biological response to the polymer. Understanding how to optimize cellular proliferation and differentiation can lead to the development of better implants that will integrate into the host’s structure and facilitate tissue regeneration.

OBJECTIVES/GOALS: The need for mechanisms of ethical discourse and guidance has increased as translational research collaborations become more complex. The goal of this project is to analyze the stakeholder engagement and ethical issues our research ethics consultation service (RECS) conducted over a two year period. METHODS/STUDY POPULATION: We conducted a retrospective review of our RECS database from 2020 to 2022. We examined the nature of the research and ethical issues of concern from consult requestors, including whether or not consults were preventative. In addition, we assessed the educational outreach conducted during that timeframe as a measure of service awareness. RESULTS/ANTICIPATED RESULTS: There was a total of 42 consults conducted over the previous year. There were a wide variety of issues related to informed IRB-related processes (31%), consent (24%), QA/QI determination (12%), authorship (10%), confidentiality (7%), diversity/inclusion (7%), grant preparation (7%). Many of the consults (n=28, 67%) included secondary issues. A few consults (n=4, 10%) were preventative, meaning that the consult was requested in anticipation or consideration of a potential ethical issue. Outreach efforts extended to a diverse array of institutional stakeholders and trainees. DISCUSSION/SIGNIFICANCE: The RECS serves numerous constituencies throughout our institution on ethical issues spanning nearly all aspects of research design, conduct, and analysis. These data highlight initiatives to increase study efficiency (in collaboration with institutional research oversight) and helps to direct educational efforts and outreach.

ABSTRACT IMPACT: Glucocorticoid steroids are commonly used despite known dose-dependent cardiovascular toxicity, yet little is known about a) how patients with other cardiovascular risk factors use glucocorticoids, and b) how risks of glucocorticoid treatment might vary depending on a patient’s baseline cardiovascular risk. OBJECTIVES/GOALS: Up to one-third of RA patients use long-term glucocorticoids (GCs) despite a known, dose-dependent association with increased risk of major adverse cardiovascular events (MACE). We aim to evaluate patterns of GC use among RA patients with other MACE risk factors (i.e. diabetes, smoking), and examine how GC use may potentiate these risk factors. METHODS/STUDY POPULATION: We used claims data from Veterans Health Administration to identify 6,090 RA patients with ≥1 rheumatology clinic visit during 2013-2017. We used logistic regression to evaluate associations between incident MACE between 2013-2018, recent long-term GC use, and 5 MACE risk factors: hypertension, diabetes, hyperlipidemia, smoking, and prior MACE. We included two-way interaction terms between GC use and each risk factor. We used a claims-based algorithm to define MACE as any of acute MI, ischemic stroke, TIA, sudden death, or coronary revascularization, between index date and 12/31/2018. We defined index date as first rheumatology visit after meeting RA diagnostic criteria, and recent long-term GC use as ≥90 days’ supply dispensed over 2 years prior to index date. RESULTS/ANTICIPATED RESULTS: Among 2,884 eligible patients,1,553 (54%) had MACE risk factors, and 97 (3%) had prior MACE (Table 1). Overall, 16% of patients recently used long-term GC, compared to 17% of patients with MACE risk factors, and 22% of patients with prior MACE. Incident MACE occurred in 308 (11%) patients, 24% of whom had recent long-term GC use. Recent long-term GC use was independently associated with increased incident MACE (Table 2). While no interaction term was statistically significant overall, differences in odds of incident MACE were seen across levels of recent GC use for several risk factors, particularly diabetes (OR 2.10, 95% CI [0.93-4.77]), tobacco use (OR 2.88, 95% CI [1.16-7.14]) and prior MACE (OR 2.41, 95% CI [0.73-7.95]). DISCUSSION/SIGNIFICANCE OF FINDINGS: Long-term GC use is common among RA patients with MACE risk factors. In this cohort, 25% of patients with incident MACE had recently used long-term GC. Long-term GC use may potentiate effects of comorbidities like diabetes and smoking, disproportionately increasing MACE risk in certain patients.

OBJECTIVES/GOALS: Pancreatic cysts are comprised of both precancerous mucinous lesions and non-mucinous lesions with minimal malignant potential. Our goal is to improve our ability to classify the type of cyst using a combination of novel radiomic features and cyst fluid proteolytic activity. METHODS/STUDY POPULATION: Preoperative pancreatic protocol CT images from 30 patients with proteolytic assay characterization, followed by surgical resection with a pathologically confirmed pancreatic cyst diagnosis between 2016-2019 will be used in this study. We will contour images using the widely available software 3D Slicer, and extract radiomic features using IBEX software. We will analyze area under the ROC curves to identify the radiomic features that best differentiate mucinous from non-mucinous cysts, and identify features to be cross validated. The predictive ability of identified radiomic features combined with proteolytic assay will be determined by performing multiple logistic regression analysis and comparing AUROC analysis. We will determine sensitivity and specificity for individual, as well as combinations of, analytes to determine the optimal classifier. RESULTS/ANTICIPATED RESULTS: We anticipate that the predictive ability, sensitivity, and specificity of utilizing radiomic features combined with proteolytic assay data will exceed the performance of any individual test. DISCUSSION/SIGNIFICANCE OF IMPACT: This work is designed to provide a predictive radiomic model that will enable us to better identify mucinous cysts that require further evaluation, and potentially prevent unnecessary surgery in other patients. Ultimately, we would like to improve the accuracy of noninvasive radiographic evaluation using radiomic markers. CONFLICT OF INTEREST DESCRIPTION: Dr. Charles Craik is a co-founder of Alaunus Biosciences, Inc.

OBJECTIVES/SPECIFIC AIMS: Using national Medicaid claims, this retrospective cohort study aims to compare the outcomes of levetiracetam (LEV) versus phenobarbital (PHB) as initial monotherapy in infants with epilepsy aged one month to one year. We primarily analyzed health services outcomes, as follows: (1) Emergency Department (ED) visits: proportion of those with at least 1 ED visit, time to first ED visit, total number of ED visits, proportion leading to inpatient admission (2) Inpatient Admissions: proportion of those with at least 1 admission, time to first admission, total number of inpatient admissions, total length of stay (3) Treatment discontinuation: proportion of those who discontinued medication, time to discontinuation, proportion of those with prescription of second antiepileptic drug (AED) (4) Cost: total cost at 1 year from first prescription. METHODS/STUDY POPULATION: The project is a retrospective cohort study utilizing Medicaid claims from 2009-2012 from all states and DC. Inclusion criteria were: (1) Epilepsy diagnosis (ICD9 345.x) at age 1 month to 1 year (2) 1 year of continuous Medicaid enrollment Exclusion criteria were: (1) Neither LEV nor PHB monotherapy as initial treatment (2) Neither LEV nor PHB prescription within 45 days of epilepsy diagnosis (3) Less than 1 year of follow-up after first LEV or PHB prescription (4) Infantile Spasms diagnosis (5) Brain Surgery (6) Death within 1 year of follow-up Using R for statistical analysis, we analyzed outcomes including ED visits, inpatient admissions, treatment discontinuation, and total cost. In this abstract, we present our preliminary bivariate analysis. RESULTS/ANTICIPATED RESULTS: Demographics and Etiology: Compared to infants prescribed PHB (n = 1954), infants prescribed LEV (n = 1248) were older (median 6 months [IQR 3-9] vs 3[2-6]; p < 0.001), more likely to be white (44.2% vs 38.9%; p < 0.05) and not Hispanic (63.5% vs 58.9%; p<0.05). There were also important differences in epilepsy etiologies (p < 0.05). For example, infants prescribed LEV were more likely to have a diagnosis of tuberous sclerosis (1% vs 0.15%) or traumatic brain injury (12.8% vs 0.56%). Health Services Outcomes: After 1 year, infants prescribed LEV had more ED visits (2 [0-4] vs 1 [0-3]; p < 0.001) but shorter inpatient length of stay when admitted (3 days [2-5] vs 3 [2-6]; p < 0.001). They were less likely to discontinue the medication (46.6% vs 64.3%; p<0.001) but more likely to have a second AED prescription (53.3% vs 43.4%; p < 0.001). Other outcomes, including total cost, were similar. DISCUSSION/SIGNIFICANCE OF IMPACT: This preliminary analysis suggests that the healthcare trajectory of infants treated with LEV and PHB differ in complex ways. In ongoing work, we are conducting a multivariable comparative effectiveness analysis of LEV versus PHB using propensity score weighting to account for observable selection bias and multiple imputation to account for missing data.

OBJECTIVES/GOALS: The COVID-19 pandemic has impacted nursing frontline professionals. The aims of this study were to explore experiences of nursing professionals in Puerto Rico during the pandemic, examine the impact on their health and provide research development opportunities enhance research capacity. METHODS/STUDY POPULATION: This interpretative phenomenological study recruited graduate nurses who participated in one in-depth semi-structured virtual interviews. Interviews were audio recorded and transcribed. The data analysis process was guided using the following steps: 1. Reading and re-reading, 2. Initial noting, 3. Developing emergent themes, 4. Searching for connections across emergent themes, 5. Moving to the next case, 6. Looking for patterns across cases, and 7. Writing up. In addition, Van Manen's thematic structure of four foundations was used as a complement to guide reflection and interpretation. Faculty and students participated throughout the process. RESULTS/ANTICIPATED RESULTS: Seven nursing professionals’lived experiences caring for Covid-19 patients were gathered. Their ages ranged from 31 to 45 and had worked between 2 and 14 years providing direct care. Themes that emerged from narrations include compassion fatigue, teamwork, working beyond clinical role, and gratification. Nurses expressed dealing with a very difficult situation, fear of being infected , or infecting my family , and working together to get through it and better help patients . Nurses also expressed feelings of anxiety and lack of institutional support. Additionally, the impact of working with patients, feeling good for being there, good or bad and support from families. DISCUSSION/SIGNIFICANCE: Nurses’narrations point to the complexities of their experiences working during the pandemic. They had to transcend usual demands even though they often lacked needed support. We must recognize the value of nursing and reflect upon changes in healthcare that are essential to move nursing forward.

OBJECTIVES/GOALS: Metabolic dysfunction-associated steatotic liver disease (MASLD) is a major public health concern due to its increasing prevalence and association with type 2 diabetes mellitus. Non-invasive magnetic resonance-based biomarkers can aid in the monitoring of disease progression and identification of patients at risk for chronic liver disease. METHODS/STUDY POPULATION: Over 600 subjects will be recruited from the San Antonio Mexican American Family Study and from a second study, which consists of (i) T2DM patients diagnosed with either MASLD or metabolic dysfunction-associated steatohepatitis (MASH) or (ii) metabolically healthy controls. Hydrogen-1 MRS and diffusion-weighted MRI (DW-MRI) will be used to measure liver fat fraction and liver stiffness biomarkers, respectively. Several potential biomarkers of liver stiffness will be evaluated in vivo using the intravoxel incoherent motion (IVIM) model for DW-MRI. To further improve the diagnostic accuracy of patients with liver fibrosis, we will integrate MRI/MRS data with relevant clinical indicators of hepatic metabolism. Results will be compared to biopsy samples to evaluate the model’s diagnostic accuracy. RESULTS/ANTICIPATED RESULTS: Based on preliminary data, we predict that IVIM will be able to accurately diagnose hepatic fibrosis in patients with MASLD, allowing it to be implemented in clinics with high-field MRI units easily. Previous studies have shown correlations between IVIM estimates and fibrosis stages, however, none included additional clinical indicators of liver disease in their models. We have already found significant differences in metabolic measurements such as fasting plasma glucose and HbA1c levels. Additionally, the use of machine learning in developing these models has shown improvements in the ability to extract features from the data. The aim is to achieve high accuracy and robustness in the staging of liver fibrosis. DISCUSSION/SIGNIFICANCE: Over 100 million people in the US are affected by MASLD. Without treatment, it progresses from hepatic steatosis to MASH, fibrosis (liver stiffening), and ultimately to hepatic cirrhosis and hepatocellular carcinoma (HCC). Continued research efforts and clinical implementation of MRI and MRS are vital in combating the growing burden of MASLD.

OBJECTIVES/GOALS: The study aims to utilize event-related potentials (ERPs) coupled with observable reports of symptoms to comprehensively understand neurological and symptomatic profile of individuals at risk for developing psychosis. The study is a short-term longitudinal design which allows for examination of course as well as structure of illness. METHODS/STUDY POPULATION: This study uses a combination of well-validated ERPs (P300, N400, ERN) and symptom data to predict variation in symptoms over time. We parse heterogeneity within a high-risk group to create innovative profiles and predict variation in course of symptoms. Data collection is ongoing (n = 35; target N = 100). Methods include a battery of ERP tasks tracking neural processes associated with attention, language processing, and executive function (P300, N400, ERN), along with assessment of symptom type and severity. Analyses include how ERPs correlate with severity of risk and symptom dimensions (positive, negative, disorganized). We examine whether individual versus global ERP aberrations (P300, N400, ERN) predict individual versus global symptom domain severity (positive, negative, disorganized), or vice versa. RESULTS/ANTICIPATED RESULTS: Symptom domain scores were elevated compared to general population on positive (M = 1.65, SD = .36), negative (M = 1.9 SD = .42), and depressive (M = 1.94, SD = .40) domains. Small to medium effect sizes emerged for P300 profile (r’s = −.001 to −.41) and ERN profile (r’s = −.03 to −.37), though small effect sizes for N400 profile (r’s = −.06 to .29). Analyses were run to determine the degree to which profiles of risk were similar: P300/ERN (r = −.09), ERN/N400 (r = −.39), and N400/P3 (r = −.20). Additional analyses suggest potential mediating effects of cognition on neural activity and symptoms. DISCUSSION/SIGNIFICANCE OF IMPACT: We use a combination of well-validated ERPs (i.e. P300, N400, ERN) with behavioral and symptom data to predict variation in symptoms over time. A “fingerprint” physiologic aberration may be exhibited within high-risk individuals and can be used as biomarkers to identify those at risk even before onset of observable symptoms.

OBJECTIVES/GOALS: To identify empiric neuropsychiatric symptom (NPS) clusters in behavioral variant frontotemporal dementia and to determine the role of early anxiety/depression on functional progression. METHODS/STUDY POPULATION: Analyses were conducted using data from the ARTFL-LEFFTDS Longitudinal Frontotemporal Lobar Degeneration (ALLFTD) study, an established consortium with an ongoing cohort study of FTD patients across 18 clinical sites which includes comprehensive cognitive, neuropsychiatric, and structural neuroimaging data. A polychoric cluster analysis was performed on subjects from the ALLFTD cohort [applewebdata%3A//044E463E-34DA-4677-9EDC-B8309D14C337#_msocom_1] with early-stage disease (N=145, male 61%, median age 62 years) in order to identify empiric NPS clusters. Cox proportional hazard regression was then used to examine the association between early affective symptoms in bvFTD and subsequent functional disabilities adjusted for age, sex, level of education, and FTLD CDR global score. RESULTS/ANTICIPATED RESULTS: We identified a four-factor model as the best fit for the data: (1) an affective cluster with prominent depression, anxiety, agitation, and irritability, (2) a disinhibited symptom cluster with prominent elation and disinhibition, (3) an obsessive symptom cluster with prominent obsessive/ritualistic behavior and hyperorality, and (4) a psychotic symptom cluster with prominent delusions and hallucinations. The hazard of developing impairments in transactions, language, self-care, meal preparation, and incontinence was significantly elevated in those with early affective symptoms (depression/anxiety). DISCUSSION/SIGNIFICANCE: In this study we show that, NPS cluster into four discrete groups: (1) affective symptoms, (2) disinhibited symptoms, (3) obsessive symptoms, and (4) psychotic symptoms. Anxiety and depression are prominent within the affective symptom cluster and are associated with accelerated functional decline in a number of domains.