bluuebunny/crossref_metadata_2025 huggingface dataset:
309 Effects of SARS-CoV-2 Variants on CD8+ T cell Epitope Diversity: Estimating Clinical Severity in the United States
Authors: Kim Grace , Elnaggar Jacob , Sevalia Maya , Nicholas Najah , Varnado Mallory , Crabtree Judy , Miele Lucio
Published: April 2024

OBJECTIVES/GOALS: Our goal was to distinguish SARS-CoV-2 CD8+ T cell epitopes of spike, membrane, and nucleocapsid products in 27 of the most frequent HLA-Aand -B alleles.We hypothesize that differences mediated by variation in SARS-CoV-2 and host HLA genetics affect the differential clinical severity and presentation of acute infection and PASC. METHODS/STUDY POPULATION: Genomic sequences of SARS-CoV-2 variants were blasted against the original Wuhan strain using Ensembl’s SARS-CoV-2 browser. We examined 16 COVID variants: 2 Alpha (B.1 and B.1.1.7), 5 Delta (AY.100, AY.25, AY.3, AY.3.1, and AY.44), and 9 Omicron (BA.1, BA.1.1, BA.2, BA.4, BA.5, BQ.1, BQ.1.1, XBB.1, and XBB.1.5), sequenced from the Louisiana patient population. cDNA sequences were translated using the Expasy tool. To predict MHC-I epitope binding, we used the Immune Epitope Database and Analysis Resource, via TepiTool utilizing the IEDB recommended default prediction and the 27 most frequent HLA-A and -B alleles. In silico peptide docking was conducted on FoldX, utilizing HLA-B*15:01 structures (n= 7) from the Protein Data Bank. RESULTS/ANTICIPATED RESULTS: CD8+ epitope conservation was estimated at 87.6-96.5% in S, 92.5-99.6% in M, and 94.6-99% for N. As the virus mutated, an increasing proportion of S epitopes experienced reduced predicted binding affinity: 70% of Omicron BQ.1- XBB.1.5 S epitopes experienced decreased predicted binding, as compared to ~ 3% and ~15% in Delta AY.100-AY.44 and Omicron BA.1-BA.5 respectively. Additionally, we identified several novel candidate haplotypes that may be susceptible to severe disease, notably HLA-A*32:01, -A*26:01, -B*58:01, and -B*53:01, and relatively protected from disease, such as -A*01:01, -A*02:01,-A*31:01, -B*15:01, -B*40:01, -B*44:03, and -B*57:01. In silico analysis of COVID peptides and HLA-B*15:01, a common allotype in the United States, largely matched predicted binding patterns. DISCUSSION/SIGNIFICANCE: To elicit long term COVID-19 immunity and prevent PASC, it is important to understand the relationship between T-cells, viral variants, and HLA genetics. This project is one of the first to explore the interaction between CD8+ epitope diversity and viral genetics for the majority of the United States population.
314 Large Language Model Approaches to Understand Differences Between Guidelines and Clinician Perception of Best Practices
Authors: Gold Carrie E. , Butler Jorie M. , Peltan Ithan D. , Facelli Julio C.
Published: April 2024

OBJECTIVES/GOALS: The Clinical Implementation stage in the translational pipeline is hampered by the tension between formal evidence and clinician perceptions. For instance, when guidelines are translated into electronic clinical decision support alerts, they are often ignored. Using advances in LLMs we present a framework to quantify these discrepancies. METHODS/STUDY POPULATION: We hypothesize that ignoring guideline-based alerts may be driven by discordances between clinical guidelines’ deterministic realities and clinician’ perception of clinical reality. Until now this has been very difficult to measure using quantitative methods. We argue that advances in Large Language Models (LLM) provide an avenue for exploring this quantitatively. Here we present the method and preliminary results comparing the responses of BioBERTT from a carefully designed set of questions when the LLM is fine-tuned using either formal guidelines or transcripts of clinicians discussing guidelines and clinical care in the parallel domain. The formal “distance” between the LLM responses is evaluated using quantitative metrics like the Hamming Distance. RESULTS/ANTICIPATED RESULTS: We present a description of the architecture used to prove or disprove our hypothesis. We will present results obtained when training the architecture with data that could be used to test the limits of our hypothesis, by fine-tuning BioBERT with diverse synthetic clinical views, either in agreement or disagreement with the formal guidelines. Results comparing sepsis guideline text with transcripts of interviews with Emergency Department clinicians discussing care practices for sepsis in the ED transcripts will also be considered. Our current emphasis is on securing a wider range of transcripts of clinicians interviewed from different clinical specialties and different clinical settings. While here we focus on clinical guidelines, the framework supports any intervention in the Clinical Implementation stage. DISCUSSION/SIGNIFICANCE: Leveraging recent advances in LLMs, we develop a framework that can quantitatively measure the differences between guidelines and clinician perception of best practices. We demonstrated the functionality of this approach using synthetic data and initiated the collection of clinician transcripts to test the framework in real clinical situations.
319 Development and Validation of an Artificial Intelligence Model to Accurately Predict Spinopelvic Parameters
Authors: Harake Edward S , Linzey Joseph R. , Jiang Cheng , Jones Jaes C. , Joshi Rushikesh , Zaki Mark , Wilseck Zachary , Joseph Jacob , Hollon Todd , Khalsa Siri Sahib S. , Park Paul
Published: April 2024

OBJECTIVES/GOALS: The correction of spinopelvic parameters is associated with better outcomes in patients with adult spinal deformity (ASD). This study presents a novel artificial intelligence (AI) tool that automatically predicts spinopelvic parameters from spine x-rays with high accuracy and without need for any manual entry. METHODS/STUDY POPULATION: The AI model was trained/validated on 761 sagittal whole-spine x-rays to predict the following parameters: Sagittal Vertical Axis (SVA), Pelvic Tilt (PT), Pelvic Incidence (PI), Sacral Slope (SS), Lumbar Lordosis (LL), T1-Pelvic Angle (T1PA), and L1-Pelvic Angle (L1PA). A separate test set of 40 x-rays was labeled by 4 reviewers including fellowship-trained spine surgeons and a neuroradiologist. Median errors relative to the most senior reviewer were calculated to determine model accuracy on test and cropped-test (i.e. lumbosacral) images. Intraclass correlation coefficients (ICC) were used to assess inter-rater reliability RESULTS/ANTICIPATED RESULTS: The AI model exhibited the following median (IQR) parameter errors: SVA[2.1mm (8.5mm), p=0.97], PT [1.5° (1.4°), p=0.52], PI[2.3° (2.4°), p=0.27], SS[1.7° (2.2°), p=0.64], LL [2.6° (4.0°), p=0.89], T1PA [1.3° (1.1°), p=0.41], and L1PA [1.3° (1.2°), p=0.51]. The parameter errors on cropped lumbosacral images were: LL[2.9° (2.6°), p=0.80] and SS[1.9° (2.2°), p=0.78]. The AI model exhibited excellent reliability at all parameters in both whole-spine (ICC: 0.92-1.0) and lumbosacral x-rays: (ICC: 0.92-0.93). DISCUSSION/SIGNIFICANCE: Our AI model accurately predicts spinopelvic parameters with excellent reliability comparable to fellowship-trained spine surgeons and neuroradiologists. Utilization of predictive AI tools in spine-imaging can substantially aid in patient selection and surgical planning.
331 Validation of a Novel CSF-Based Biomarker of Mitochondrial Function
Authors: Abeykoon Dhanushki , Wang Xiaowan , Novikova Lesya , Ranjan Amol , Gabrielli Alexander , Swerdlow Russell
Published: April 2024

OBJECTIVES/GOALS: Determine the exosome mitochondrial DNA (mtDNA) copy number in cerebrospinal fluid (CSF) as a measure of neuronal mitochondrial integrity in patients with subarachnoid hemorrhage (SAH). Determine the patterns of beta amyloid and tau protein biology in CSF of SAH patients and correlate those measures with the clinical status of the SAH patients. METHODS/STUDY POPULATION: The CSF is collected from SAH patients undergoing ventriculostomy-based continuous CSF drainage. Adults from all ethnicities and sex are included in this study. The exosomes are isolated from CSF samples using a precipitation method and particle count and size are measured using NanoSight. The DNA is extracted using an exosomal DNA isolation kit (XCF kit). The CSF mtDNA copy number is measured using digital drop PCR with mitochondrial DNA primers. The levels of beta-amyloid (a-beta-40 and -42) and tau protein in CSF are measured using a sensitive ELISA-based assay. A quantitative evaluation of mitochondrial DNA copy number, clinical status of the SAH patients and beta amyloid, and tau protein levels will be conducted and reported. RESULTS/ANTICIPATED RESULTS: Preliminary results of four CSF samples showed similar patterns in CSF exosome particle number, particle size and exosomal mtDNA copy number in relation to samples from the admission day. Particle number decreased with time while particle size increased. More patient samples will be analyzed to confirm the patterns. We anticipate that mtDNA copy number will correlate with brain beta-amyloid and tau protein levels. Moreover, we anticipate that the clinical status of the SAH patients will associate with the mtDNA copy number. We specifically predict that higher mtDNA copy number levels will correlate with better clinical outcomes. DISCUSSION/SIGNIFICANCE: Mitochondrial function is critical to brain health, but we lack effective ways to monitor this parameter. Here we focus on a CSF based biomarker, exosome-derived mtDNA, which is intended to reflect the integrity of brain mitochondria. As bioenergetic metabolism influences beta amyloid and tau biology, predicting those levels are important.
334 Identification of novel plasma protein biomarkers for diagnosis and prediction of Alzheimer’s disease in African Americans
Authors: Kuchenbecker Lindsey , Reddy Joseph S , Carrasquillo Minerva M
Published: April 2024

OBJECTIVES/GOALS: To identify novel panel of plasma protein biomarkers to improve prediction and diagnosis of Alzheimer’s disease (AD) for African Americans (AA), who are at greater risk of developing AD compared to non-Hispanic White individuals but are underrepresented in AD research. METHODS/STUDY POPULATION: Pre-existing plasma samples from 460 AA individuals with clinical diagnoses of AD, cognitively unimpaired (CU), mild cognitive impairment (MCI), or dementia with Lewy bodies (DLB) will undergo untargeted proteomics using the SomaScan assay, where modified single stranded DNA aptamers bind to protein targets which are quantified by DNA microarray. Protein expression levels will be compared between diagnostic groups to identify differentially expressed proteins. Additional clinical, genetic, and lifestyle factors will be compared with protein expression when available. Proteins of interest, identified by differential protein expression analysis results, will be included in receiver operating characteristic analyses to identify the optimal set of proteins for diagnostic classification. RESULTS/ANTICIPATED RESULTS: A pilot experiment utilizing plasma from 40 individuals identified multiple differentially expressed proteins (DEPs) between AD and non-AD groups. Eight proteins were nominated from the differential protein analysis into a receiver operating characteristic (ROC) analysis based on pvalue and previous implication in AD genome wide association studies. Proteins involved in microglial activation, neuronal adhesion, cell proliferation, and innate immunity were nominated. The ROC model achieved 100% classification accuracy of AD and CU groups using age, sex, and the eight nominated proteins. It is expected that there will be more significant associations when utilizing the full cohort of 460 AA and that DEPs between AD, CU, MCI, and DLB will be identified. DISCUSSION/SIGNIFICANCE: The nomination of a novel panel of plasma biomarkers developed from an AA cohort will directly serve the AA community by improving access to an early and accurate diagnosis of AD. Access to improved prediction and diagnosis will likely improve disease management, thus improving patient outcomes and decreasing burden on families and caregivers.
338 Non-occupational herbicide and VOC exposures detected in dogs with multicentric lymphoma: a model for human non-Hodgkin lymphoma
Authors: Tindle Ashleigh , Kreuger Lauren , Trepanier Lauren
Published: April 2024

OBJECTIVES/GOALS: The objective of this study was to determine whether pet dogs with multicentric lymphoma (ML), a spontaneous, immunocompetent model for human non-Hodgkin lymphoma (NHL), are exposed to higher concentrations of herbicides and volatile organic compounds (VOCs) compared to matched unaffected control dogs. METHODS/STUDY POPULATION: We are prospectively enrolling dogs with ML within a single high-risk breed, the boxer dog, along with age-matched control boxers sampled within the same season. We are measuring urinary concentrations of the herbicides glyphosate (in Roundup®) and 2,4-D, as well as stable urinary metabolites of the VOCs benzene and 1,3-butadiene. To assess the genotoxic potential of herbicide and VOC exposures, we are using reverse dosimetry to estimate plasma exposures, and exposing healthy canine PBMC’s to these concentrations of herbicides and VOCs in vitroto assess double stranded DNA damage using the Comet Chip assay. RESULTS/ANTICIPATED RESULTS: Preliminary data show significantly higher benzene exposures, measured by the stable benzene metabolite PHMA, and significantly higher 2,4-D exposures at the time of diagnosis in cases versus controls. All dogs had measurable exposures to 1,3-butadiene (measured as its stable metabolite DHBM) and glyphosate.In vitro results show significant genotoxicity thresholds of 0.1 uM for both glyphosate and 2,4-D in dog lymphoid cells. To date, these predicted plasma exposures have not been reached in vivo in boxer dogs with ML or unaffected control boxers. DISCUSSION/SIGNIFICANCE: Canine multicentric lymphoma resembles human NHL and is a potentially useful model of non-occupational chemical risk for NHL in people. The goal of this research is to identify potentially preventable non-occupational chemical risk for lymphoma and support evidence-based remediation strategies to decrease lymphoma risk in both humans and dogs.
340 Anti-CD20 attenuates lung humoral and cellular immune responses: implications for gene therapy in Cystic Fibrosis
Authors: Clark Robert , Rabito Felix , Munyonho Ferris , Remcho Parks , Kolls Jay K.
Published: April 2024

OBJECTIVES/GOALS: Re-administration of inhaled gene therapies has the potential to overcome low correction efficiencies and limiting immune responses observed in previous trials of gene therapy for Cystic Fibrosis. We therefore tested the hypothesis that pre-treatment with a B-cell depleting α ± CD20 antibody would permit vector re-administration. METHODS/STUDY POPULATION: We first selected Adenoviral (Ad) vectors to study initially due to their well-known ability to elicit potent immune responses. Mice were dosed with a depleting α ± CD20 antibody or isotype control 2 days prior to delivery of Ad-Luc. 4 weeks later, mice were euthanized to assess the development of anti-vector immune responses. Flow cytometry and single-cell RNA sequencing were used to evaluate the development of lung-resident memory cells. Serum and airway antibody responses were assessed by ELISA. After 4 weeks, mice were dosed with Ad-LacZ and euthanized 3 days later to assess efficiency of second-round gene transfer by Î²-galactosidase activity assay. Similar methods were used in a pilot experiment with Adeno-associated virus vector (AAV), but with euthanasia 3 weeks after secondary gene transfer. RESULTS/ANTICIPATED RESULTS: Delivery of Ad vectors leads to the development of lung-resident memory B and T-cells. The depletion of B-cells prior to first-round vector delivery attenuated airway T-cell infiltration and serum IgG production, abrogated mucosal IgG and IgA production, and completely rescued secondary gene transfer. Genetically modified mouse models suggest secreted antibodies are critical in prevention of vector redosing. AAV vectors were found to be less immunogenic than Ad vectors, with only partial reduction of second-round gene transfer. However, anti-CD20 provided no benefit for AAV redelivery. DISCUSSION/SIGNIFICANCE: Mucosal humoral immunity is critical in preventing re-administration of Adenoviral vectors. Impairment of B-cell responses by α ± CD20 treatment prior to vector delivery allows re-administration and may help overcome low efficiencies of CF gene therapy. AAV vectors may be less susceptible to neutralization by pre-existing immunity.
342 Impact of the Endothelial Nucleotidase on Thrombosis
Authors: Gordon Kyle , Blanco Oscar Bermeo , Covarrubias Roman , Gumina Richard J.
Published: April 2024

OBJECTIVES/GOALS: Contrary to current dogma, prior work has suggested that in humans, SNP-determined endothelial cell reduction in CD39 expression associates with a diminished risk for venous thromboembolism. Our objective was to examine the impact of endothelial cell (EC) CD39 expression on arterial thrombosis that replicates human data. METHODS/STUDY POPULATION: We generated a novel CD39 cell-specific conditional knockout mouse line for endothelial cells (EC-cKO: Tie2-Cre+; cd39flox/flox versus WT: Tie2-Cre-; cd39flox/flox). We validated the knockout of expression of CD39 on EC using FACS analysis and measured EC CD39 activity using the Kinase Glo ATP hydrolysis assay on magnetically sorted EC. We then used a standard FeCl3 carotid injury model to evaluate time to arterial thrombosis in vivo by measuring time to occlusion tracked via a Doppler flow probe on the exposed vessel. RESULTS/ANTICIPATED RESULTS: FACS analysis revealed a specific 97% knockout of CD39 expression on EC (p<0.001) but not on other cells within the vasculature. There was also significant reduction in ATP hydrolysis (81%; p=0.019) in EC-cKO mouse EC versus WT. We next examined the time to arterial thrombosis. EC-specific conditional knockout of CD39 exhibited a significant prolongation in time to thrombosis compared to WT (WT: 8.28 minutes +/- 0.82; EC-cKO: 11.92 minutes +/- 1.34; p=0.024). Analysis of carotid blood-flow revealed that EC-cKO and WT mice had similar baseline blood flow velocity (p=0.51), but after vessel injury with FeCl3, EC-cKO mice exhibited a 16% increase maximal flow velocity relative to baseline compared to WT (p<0.001), as well as a 19% increase at 2-minutes post-injury in comparison to EC-WT mice (p<0.001). DISCUSSION/SIGNIFICANCE: Our findings demonstrate that CD39 activity plays a role in modulating arterial thrombosis and blood-flow regulation within the vasculature. These findings exemplify the therapeutic potential of modulating endothelial CD39 activity, as well as the potential for using SNPs within the gene coding for CD39 as a cardiovascular disease marker.
346 Multimodal assessment of sleep in individuals with chronic post-concussive symptoms: A Pilot Study
Authors: Wong Caitlyn Emma , Koike Seiji , Luther Madison , Scatena Avery , Dennis Laura , Yamamoto Erin A , Khambadkone Seva , Garavatti Emily , Piantino Juan
Published: April 2024

OBJECTIVES/GOALS: Weaimed to compare subjective and objective sleep in individuals with chronic post-concussive symptoms. We hypothesized an association between self-reported sleep quality and objective sleep parameters, which is different for concussed and control cohorts. METHODS/STUDY POPULATION: 28 individuals with chronic post-concussive symptoms and 13 age-matched controls (no concussion history) completed the ISI, PSQI, PROMIS Depression, Anxiety, Stress and Cognitive questionnaires at enrollment. Objective sleep parameters were obtained for a minimum of 7 days and up to 30 days with a validated sleep monitoring device placed under the subject’s bed (Emfit). For each night, raw activity data per minute were analyzed to determinein-bed, sleep, wake, andout-of-bedtimes. These measures were used to calculate total sleep time (TST), sleep onset latency (SOL), and wake after sleep onset (WASO) for each night. RESULTS/ANTICIPATED RESULTS: Concussed individuals reported worse sleep with PSQI and ISI scores significantly higher than controls. They also showedsignificant associations between PSQI and Dep ression, ISI and Depression, and ISI and Anxiety scores. There was no difference between objective sleep parameters in the concussed and control cohorts (in-bed/sleep/wake/out-of-bed times, TST, SOL, and WASO). Instead, higher PSQI, ISI, Depression, Anxiety, and Stress scores (greater symptom burden) were all associated with later sleep times, where as higher Cognitive scores (greater cognitive function) were associated with earlier sleep times, regardless of group status. DISCUSSION/SIGNIFICANCE: Concussed individuals report worse subjective sleep but no differences to controls when objectively assessing sleep. Depression/anxiety, and not concussion status, determine objective sleep parameters. Psychiatric comorbidities should inform the treatment of post-concussive sleep disturbances.
348 Alexithymia impacts vulnerability for cognitive decline in healthy elders via frontal lobe connectivity during response inhibition, especially in women
Authors: Otteman Christian , Nielson Kristy A.
Published: April 2024

OBJECTIVES/GOALS: This project aimed to examine the impacts of biological sex and alexithymia on frontal lobe connectivity in executive functioning (EF)-related neural networks during successful inhibition as a means to index vulnerability for future cognitive decline. METHODS/STUDY POPULATION: Healthy, cognitively intact older adults (n=43, 33 female, Mage=79) completed the 20-item Toronto Alexithymia Scale (TAS-20) and the stop-signal task in this study. We used electroencephalography (EEG) source estimation to investigate EF-related frontal connectivity during successful inhibition in stop-signal task trials. Connectivity was measured in bilateral frontal ROIs relevant to inhibition using time series correlations over the N200 (186-350ms) and P300 (340-616ms) time windows, associated with the inhibitory subprocesses of conflict processing and performance evaluation, respectively. RESULTS/ANTICIPATED RESULTS: Those with higher alexithymia, as measured by the difficulty identifying feelings (DIF) facet of the TAS-20, had lower right anterior cingulate cortex (ACC)-left superior frontal gyrus (SFG) connectivity in the P300 window, suggesting impaired performance evaluation. Further, in females specifically, those with higher DIF had greater right inferior frontal gyrus (rIFG)-bilateral ACC connectivity in the N200 window than those with lower DIF, suggesting greater resources were allocated for conflict processing and inhibition. Right ACC-rIFG connectivity also correlated with better stop accuracy and faster stop-signal reaction time, supporting this network’s role in successful inhibition. DISCUSSION/SIGNIFICANCE: Overall, during successful inhibition, higher DIF was associated with reduced performance monitoring efficiency as well as greater resource allocation for conflict processing during motor stopping in women only. Thus, alexithymia (via DIF) may exacerbate age-related EF dysfunction and risk for future cognitive decline, especially for females.
354 Brain Structural Alterations in Metabolically Healthy and Unhealthy Obesity: A Quantitative Comparison Using Coordinate-Based Meta-Analysis
Authors: Abazid Leen F. , Kotkowski Eithan , Franklin Crystal G. , Woosley Mary D. , Garrett Amy S. , Fox Peter T.
Published: April 2024

OBJECTIVES/GOALS: The primary research goal was to identify brain alterations reliably associated with obesity using coordinate-based meta-analysis. A secondary goal was to compare brain alterations in metabolically healthy (MHO) and unhealthy (MUO) obesity. METHODS/STUDY POPULATION: Source data were peer-reviewed studies reporting locations of gray-matter alterations in group-average, case-control contrasts (obese vs. non-obese) cohorts, performed in a whole-brain, voxel-wise manner. Both voxel-based morphometry and voxel-based physiology studies were included. Three coordinate-based meta-analyses were performed: Pooled (MUO + MHO), MHO, and MUO. RESULTS/ANTICIPATED RESULTS: Thirty-two studies reporting a total of 50 case-control contrasts (MHO, 23; MUO, 27) met inclusion criteria, representing 3,368 participants (obese, 1,781; non-obese, 1587). The pooled analysis yielded 8 cerebral foci (3 nuclear, 5 cortical) in regions implicated in reward-seeking, cognitive, and interoceptive behaviors. MHO yielded 7 cerebral foci (4 nuclear, 3 cortical), partially overlapping Pooled results, with similar behavioral loadings. The MUO pattern was distinct, with 3 cerebellar and 1 occipital foci. DISCUSSION/SIGNIFICANCE: Brain alterations occurred reliably in obesity. The dominant pattern (Pooled & MHO) involved cerebral reward-system circuits, evident even in metabolically healthy obesity. Cerebellar alterations occurred exclusively in metabolically unhealthy obesity, a pattern previously reported in metabolic syndrome.
366 Implementation of COPD Clinical Practice Guidelines with Use of Telehealth
Authors: Raghavan Deepa , Sonya Sanders , Williams James , Bailey Danielle , Bartnik Mary , Kirchner JoAnn
Published: April 2024

OBJECTIVES/GOALS: Studies to improve uptake of Chronic Obstructive Pulmonary Disease Clinical Practice Guidelines (COPD CPG) have yielded inconsistent results. We hypothesized that using implementation science would facilitate rigorous site ‘diagnosis’, and promote effective contextual tailoring of COPD CPG, while piloting the use of telehealth for this. METHODS/STUDY POPULATION: The study was conducted in two Veterans Affairs primary care clinics located in a small sized city. A detailed formative evaluation was conducted using key informant interviews (with VA staff and veterans with COPD who received care at this location) and quantitative data. Multidisciplinary stakeholder group was engaged and strategies to address the determinants identified through the previous step were identified. Telehealth was strongly encouraged as the primary modality for implementing the COPD CPG and we are collecting pilot data on this. Tele-Facilitation, used as the meta-strategy was employed in conjunction with other strategies such as develop/distribute educational materials, tailor strategies, change record systems and revise professional roles. RESULTS/ANTICIPATED RESULTS: Primary Care at the VA is provided by Patient Aligned Care Teams (PACT-teams), where each team consists of multiple health professionals to provide collaborative care to the patient. Discussions with the multidisciplinary stakeholder team suggested that any implementation effort primarily focused on physician and nursing efforts was unlikely to succeed due to competing demands. A pharmacy-centric model that allowed for the PACT-team clinical pharmacist to address most of the COPD CPG (inhaler technique education/assessment, inhaler choice optimization, COPD-specific patient education, spirometry use, smoking and immunization) was developed and implemented with incorporation of telehealth (video visits and telephone). We will present pilot implementation outcomes using RE-AIM framework elements. DISCUSSION/SIGNIFICANCE: This use of implementation science to implement COPD CPG and novel use of telehealth has enormous potential for impact. Increasing reach/adoption by targeting primary care practices can help permeate quality care to the underserved population. This data will allow us to explore generalizability through wider scale implementation studies.
378 Hydroxypropyl beta cyclodextrin barrier prevents respiratory and eye viral infections
Authors: Asante Isaac , Lu Angela , Humayun Mark , Louie Stan
Published: April 2024

OBJECTIVES/GOALS: Susceptible mucocutaneous membranes of the eye and nasal cavity are easily infected by viruses leading to pink eye or respiratory infections whose direct cost has been estimated as $16 billion annually in the United States. We have developed a novel and effective barrier that will be agnostic to variants enveloped viruses like coronaviruses. METHODS/STUDY POPULATION: We evaluated the efficacy of hydroxypropyl cyclodextrin barrier in preventing respiratory coronavirus infections using 25 humanized angiotensin converting enzyme-2 receptor (hACE-2) mice under a BSL3 laboratory setting. We have shown the barrier is safe and efficacious in preventing coronavirus infections in in vitro respiratory cell lines. We instilled 10 uL aliquot of the barrier into the nostril of the mouse 30 minutes before exposing them to a 10uL titer containing 10,000 plaque forming units of the SARS-CoV-2 delta variant. The control mice received the SARS-CoV-2 infection but not the barrier. The mice were observed for 5 days after which they were sacrificed. We analyzed the lungs and nasal palates for viral load using reverse transcription-polymerase chain reaction. RESULTS/ANTICIPATED RESULTS: We observed our barrier to be effective in preventing SARS-CoV-2 delta variant infection in hACE2 mice models. The lungs and nasal secretions of treated mice were less infectious with lower viral load than the control mice. The lungs of treated mice showed decrease in IFN gene expression and many cytokines and chemokines that regulate virally induced inflammatory responses such as IL-1b, IL-8, CXCL9, CXCL10, and the CCLs. We observed the plasma Angiotensin I and Angiotensin II decreased with barrier treatment, correlating with the viral load observed in the lungs. These peptides may be useful biomarkers for monitoring viral load within the lungs of virally infected individuals. DISCUSSION/SIGNIFICANCE: This supports the barrier’s efficacy to reduce transmission and prevent infections of SARS-CoV-2. This easy to use barrier can augment the mucocutaneous layers of the eye and nasal cavity. Our agnostic barrier will reduce the economic and public health burden of seasonal respiratory and eye viral infections and their related deaths amongst the public.
388 Prototyping a mobile phone application for Chimeric Antigen Receptor (CAR) T-cell therapy patient monitoring and data collection post-discharge
Authors: Kay Shayna , Oyiliagu Chimaobi , Ali Amena
Published: April 2024

OBJECTIVES/GOALS: Research objectives include prototyping a mobile phone application that allows physicians to monitor CD19-directed CAR T-cell therapy patients remotely after discharge. This app will also enable standardized data collection across different centers that provide CAR-T cell therapy and allow for the harmonization of follow-up protocols. METHODS/STUDY POPULATION: Literature review and semi-structured interviews with patients, clinical coordinators, and other experts in the field will be used to determine what parameters must be included in the mobile application prototype to effectively monitor the side effects of CD19-directed CAR T-cell therapy. The mobile phone application will be designed using process mapping to integrate data from self-reporting and wearable technologies, including the Garmin smart watch. Figma will then be used to develop new screens based on an existing patient monitoring app for Allogeneic Stem Cell Transplant follow-up. Finally, a preliminary feasibility study will be conducted to collect feedback on the app prototype from CAR T-cell therapy patients, providers, and stakeholders. RESULTS/ANTICIPATED RESULTS: The anticipated results of this study include an app prototype that will include the functionalities required to monitor patients for adverse effects of CD19-directed CAR T-cell therapy. This will include the parameters that will be recorded or measured using a combination of self-reporting, a reliable body temperature sensor, and the Garmin watch which monitors basic vitals, activity, and sleep. Additional parameters may be added during the stakeholder co-design process. The app prototype will include a physician interface where doctors can monitor their patients and will be alerted if they require further physician assessment. It is expected that the app will provide standardized monitoring of patients when they are discharged from the hospital after receiving CAR T-cell therapy. DISCUSSION/SIGNIFICANCE: This app will allow physicians to monitor patients for general follow-up and adverse effects, including cytokine release syndrome and neurotoxicity. Future studies may utilize this app to develop best practices for harmonizing CAR-T follow-up protocols across Canada.
396 Unraveling the Immunological Basis of Lobular Involution in Breast Cancer Development
Authors: Lue Jaida , Radisky Derek , Sherman Mark , Degnim Amy , Windam Stacey , Curtis Morian , Stallings-Mann Melody
Published: April 2024

OBJECTIVES/GOALS: Reveal common immune mechanisms in dysregulated age-related lobular involution (ARLI) and post-partum lobular involution (PPLI) to understand their link to increased breast cancer risk, challenging the traditional view of their distinctiveness. Ultimately, to improve breast cancer risk assessment and personalized prevention METHODS/STUDY POPULATION: The Mayo Clinic Benign Breast Disease (BBD) cohort comprises of ~20,000 women with benign biopsies, including ~1000 women with sequential benign biopsies. Lobular involution (LI) status was assessed by selecting perimenopausal women, ages 45-55, with sequential biopsies, comparing acini number and lobule size between initial and subsequent biopsies. NanoString IO360/ BC360 RNA profiling identified differentially expressed genes associated with dysregulated LI. Using multiplex immunofluorescence (mIF), I'll analyze and spatially map immune biomarkers related to dysregulated ARLI and PPLI in BBD tissue from perimenopausal women who did or did not go on to develop breast cancer, assessing the commonality of ARLI and PPLI markers and exploring their potential as risk biomarkers for breast cancer. RESULTS/ANTICIPATED RESULTS: Preliminary findings link patients who display dysregulated ARLI with an increased breast cancer risk and identify vital PPLI biomarkers in perimenopausal women. I expect the biopsies of women who developed post-menopausal breast cancer (PMBC) and post-partum breast cancer (PPBC) to exhibit elevated levels of dysregulated ARLI immune biomarkers and PPLI biomarkers. Spatially mapping these markers promises to provide a more comprehensive understanding of their interactions, potentially revealing common immunological pathways. These findings could transform our current paradigm of ARLI and PPLI as distinct processes and demonstrate their interconnection in shaping breast cancer risk. DISCUSSION/SIGNIFICANCE: PMBC and PPBC dominate majority of breast cancer cases. Both involve activation of the understudied process of lobular involution, which has been shown to have pro-tumorigenic traits. Elucidating these mechanisms will aid more efficient risk stratification and personalized prevention to reduce incidence and mortality of breast cancer.
399 Source Localization of Burst Suppression in Post-Cardiac Arrest Patients
Authors: Velasquez Gerardo Hernandez , Kandhare Pravinkumar , Zhou Bo , Talbott Jason F , Calabrese Evan , Otero Matheus , Hemphill J. Claude , Westover M. Brandon , Lee Jong Woo , Amorim Edilberto
Published: April 2024

OBJECTIVES/GOALS: Burst suppression is a neurophysiological marker associated with severe hypoxic-ischemic injury following cardiac arrest. The goal of this study is to identify the anatomical regions of the brain associated with burst suppression post-cardiac arrest. METHODS/STUDY POPULATION: 86 comatose patients post-cardiac arrest admitted to the neurological-ICU from Massachusetts General Hospital and Brigham and Women’s Hospital were included in this study. EEG data after return of spontaneous circulation were preprocessed and artifact was rejected. Burst segments were extracted for source localization analysis from epochs with burst suppression. Four bursts for each patients were manually selected. The source of the bursts were obtained using the Champagne algorithm and mapped on the Desikan-Killiany atlas. The source for each burst was defined as any region of interest (ROI) with power > = 75th percentile relative to all ROIs. The power of the bursts at each source was correlated with the burden of brain injury measured using apparent diffusion coefficient (ADC) per ROI. RESULTS/ANTICIPATED RESULTS: 48 (56%) patients had burst suppression. 5 (10.4%) of patients with burst suppression were independent at the time of hospital discharge. Preliminary analyses was performed on 6 patients (24 bursts in total). ROI’s determined to be sources in a majority of the burst (>=13) were bilateral superior frontal, rostral middle frontal, parstriangularis precentral, superior parietal, inferior parietal, right post central, superior temporal, lateral occipital, and left middle temporal ROI. A lower mean ADC intensity was associated with a higher EEG power in the bilateral superior frontal (r = -0.80, p < 0.0001; r = -0.677, p < 0.001, respectively), left superior parietal (r = -0.53, p = 0.009), left middle temporal (r = -0.43, p = 0.042) ROI. DISCUSSION/SIGNIFICANCE: The source of bursts in patients post-cardiac arrest experiencing burst suppression is not well defined. This study will improve our understanding of how burst suppression is a measure of cortical injury, how it may relate to the burden of injury found on ADC imaging, and patient outcomes.
14 Characterization of Xylazine-Related Overdose Deaths in Maryland (2020-2022)
Authors: Wang Erin , Nestadt Paul
Published: April 2024

OBJECTIVES/GOALS: Xylazine is a strong sedative and fentanyl contaminant which has been increasingly detected in drug overdose deaths in Maryland. The goal of this project is to analyze the demographic characteristics and time trends of xylazine-related overdose deaths (XROD) in Maryland from 2020-2022. METHODS/STUDY POPULATION: This cross-sectional study utilizes the Maryland medical examiner's autopsy reports from 2020-2022. These reports include every death in the state that was investigated by the medical examiner, with demographic and toxicological data showing the presence of various substances at the time of death. An XROD was defined as someone who died from drug overdose and had a positive serum xylazine test at time of death. Demographic characteristics and time trends for XROD were analyzed. Multivariable logistic regression modeled associations between demographic variables and the presence of other substances with XROD. RESULTS/ANTICIPATED RESULTS: A total of 1,509 people died from XROD, of which the mean age was 44.4 years and 73.3% were male. The majority were White (57.6%), 39.2% were Black, and 3.2% identified as another race. Over 99.9% of individuals who died from XROD tested positive for fentanyl. XROD peaked in January 2021 and has been trending downwards since then. Adjusted multivariable logistic regression revealed that White individuals had greater odds of XROD relative to Black individuals (OR=1.22, 95% CI=1.07-1.37), and adults aged 30-45 years had higher odds of XROD relative to adults over age 60 (OR=1.26, 95%CI=1.04-1.54). Individuals who used fentanyl had higher odds of XROD relative to those who did not use fentanyl (OR=327.4, 95%CI=46.0-2331.3). DISCUSSION/SIGNIFICANCE: This study demonstrates that middle age, White race, and fentanyl use are associated with xylazine-related overdose deaths in Maryland. Efforts to reduce xylazine-related mortality in the state should address the unique social and geographic factors that influence substance use in this population.
406 A CTS Team Approach to Identifying Risk of Neonatal Hypoglycemia and its Relationship with Endothelial Dysfunction
Authors: Mahadevan Aditya Devidas , Pruitt Jennifer , Parker Leslie A. , Jones Helen
Published: April 2024

OBJECTIVES/GOALS: Neonatal hypoglycemia is seen in 65% of maternally diabetic pregnancies, and can lead to severe neurological damage. Neonatal glycemia may also be an indicator of placental function in these pregnancies. The purpose of this study is to identify patterns of neonatal glycemia, and associated endothelial dysfunction, by maternal diabetes subtype. METHODS/STUDY POPULATION: Pregnancies with maternal Type 1 (T1DM), Type 2 (T2DM), and gestational diabetes mellitus (GDM) are being enrolled. Maternal hemoglobin A1c (HbA1c) and umbilical cord insulin/glucose are being collected from 20 pregnancies in each group, 10 of which also undergo placental/umbilical cord tissue collection. Following delivery, neonatal blood glucose levels are also collected every 3-4 hours (4+ measurements) to determine rate of glycemic change. Linear regression modeling will be used to determine associations with placental and umbilical endothelial RNA expression, umbilical cord insulin levels, and maternal HbA1c within each diabetic subtype and between normoglycemic and hypoglycemic neonates. Endothelial gene expression will be compared using paired t-tests with Benjamini-Hochberg correction. RESULTS/ANTICIPATED RESULTS: Thus far, 5 T1DM, 10 T2DM, and 13 GDM samples have been collected. Gestational age at delivery and birth weight were similar between groups (38.1 ± 1.05 weeks; 3.6 ± 0.59 kilograms) and delivery method is evenly distributed (Cesarean section or vaginal delivery). Currently, with limited cohort size, no association is evident between maternal HbA1c and umbilical cord glucose/insulin (p=0.114) or neonatal hypoglycemia diagnosis (p=0.674) when controlled for gestational age and infant birthweight. We hypothesize that, with pending analyses, maternal HbA1c and umbilical cord insulin levels will correlate negatively with the rate of neonatal glycemic change, and positively with the level of inflammatory and angiogenic transcription identified in placental and umbilical endothelium. DISCUSSION/SIGNIFICANCE: Characterization of postnatal glucose control is key to prognosis and risk stratification of infants of diabetic mothers. Understanding placental response to glucose, as well as sequela in the fetal endothelium, is also critical to understanding the pathogenesis of neonatal hypoglycemia and other adverse outcomes of diabetic pregnancy.
409 Automated Prediction of Bone Volume Removed During Cortical Mastoidectomy Using Deep Learning
Authors: Nagururu Nimesh , Sahu Manish , Munawar Adnan , Barragan Juan Antonio , Ishida Hisashi , Galaiya Deepa , Taylor Russell , Creighton Francis
Published: April 2024

OBJECTIVES/GOALS: Patient-specific definition of extent of surgical excision is foundational to the safety offered by computer assisted interventions. Consequently, this study aims to develop a pipeline for automated segmentation of bone removed during cortical mastoidectomy, a technically complex otologic surgery. METHODS/STUDY POPULATION: A simulator, previously developed in our lab, allows fully immersive simulation of mastoidectomy using segmented temporal bones generated from CT data. Using the simulator, one attending surgeon will perform three trials of mastoidectomy on 20 different temporal bones. From the simulator we will obtain data on the volume of bone removed for a specific anatomy, averaged between trials. No new U-net (nnU-net), an open-source three-dimensional segmentation network, will then be trained to predict the volume of bone removed using segmented pre-operative CT imaging. Segmentation accuracy will be evaluated with the Dice coefficient, modified Hausdorff distance (mHD), sensitivity and specificity. RESULTS/ANTICIPATED RESULTS: We expect the mean pairwise Dice coefficient to be high indicating relative similarity of volume removed between trials. Moreover, we predict that following five-fold cross-validation the best model will result in a Dice coefficient, mHD, sensitivity, and specificity indicative of volume removed predictions consistent with surgeon-generated data. Finally, given that network training will penalize overlap of the predicted excised bone segment and previously segmented anatomic structures, we expect that no critical anatomical structures will be marked as tissue removed. DISCUSSION/SIGNIFICANCE: We hope to show that deep learning architectures can accurately predict bone removed during mastoidectomy. These predictions can be used for preoperative planning, as clinical endpoints in surgical simulators, or be used in conjunction with surgical robots, all ultimately improving patient safety.
412 Synergistic Targeting of Lysine-specific demethylase 1 (LSD1) and MAPK Signaling: A Mechanism-Guided Therapeutic Approach for Glioblastoma (GBM)
Authors: Stitzlein Lea , Adams Jack , Luetzen Matthew , Singh Melissa , Su Xioaping , Lu Yue , Gumin Joy , Lang Frederick , Chandra Joya
Published: April 2024

OBJECTIVES/GOALS: LSD1 is a histone demethylase important in GBM regulation. Our goal is to design a therapeutic strategy for LSD1 inhibitors to meet clinical needs in GBM. Despite the abundance of LSD1 inhibitors, resistance emerges in GBM mouse models. We aim to understand the relevance of proliferative signaling pathways, such as MAPK, in LSD1 inhibitor resistance. METHODS/STUDY POPULATION: Following LSD1 knockdown in GBM cells, we determined differentially expressed genes using RNA-seq and gene set enrichment analysis (GSEA). Kinase signaling processes enriched for LSD1 expression were identified. Utilizing western blot, we assessed LSD1’s impact on MAPK signaling in patient-derived GBM stem cells (GSCs) and pediatric high-grade glioma cell models. Pharmacological evaluation of LSD1 involved five inhibitor candidates. Additionally, we explored LSD1 inhibition in combination with brain penetrant kinase inhibitors, osimertinib and ulixertinib, directed against the epidermal growth factor receptor (EGFR) and MAPK, respectively. The treatment combinations were assessed at multiple concentrations and analyzed using SynergyFinder. RESULTS/ANTICIPATED RESULTS: Pharmacological LSD1 inhibition after 24 hours induced increased phosphorylated ERK1/2 across multiple glioma cell lines. Concurrent LSD1 and EGFR/MAPK inhibition demonstrated improvedin vitro efficacy compared to individual agents. Notably, the combination of Iadademstat (ORY-1001) and osimertinib demonstrated the highest synergy score of 37.2 using the bliss synergy model in the GSC17s. Furthermore, 11 out of the 12 combination treatments tested had a synergistic relationship, with bliss synergy scores greater than 10. DISCUSSION/SIGNIFICANCE: Our study addresses the pressing need for novel therapeutic strategies in GBM. We leveraged pharmacological tools of LSD1 inhibition to determine how they could be used most effectively, revealing kinase inhibition as a promising strategy with demonstrated in vitro efficacy. Future efforts will focus on validating these findingsin vivo.