bluuebunny/crossref_metadata_2025 huggingface dataset:
215 Impact of Maternal Diabetes on Neonatal Body Composition, Energy Homeostasis and Hypothalamic Salivary Gene Expression
Authors: Azuma Dara , Penner Yvette , Kaneko-Tarui Tomoko , Mahmoud Taysir G. , Catalano Patrick , OTierney-Ginn Perrie , Maron Jill L.
Published: April 2022

OBJECTIVES/GOALS: Infants of diabetic mothers (IDMs) may exhibit decreased oral intake, requiring nasogastric feedings and prolonged hospitalization. We hypothesize that increased insulin exposure and resulting overgrowth in utero disrupts hypothalamic regulation of food intake, correlates to body composition and impacts feeding in IDMs. METHODS/STUDY POPULATION: Infants born at â‰¥ 35 weeks gestation to mothers with gestational or type II diabetes (IDM cohort), and normoglycemic mothers (control cohort) were recruited. Infants born to mothers with Type I DM or preeclampsia and with a history of intrauterine growth restriction, opioid exposure, or major congenital anomalies were excluded. Salivary expression of known hunger signaling genes 5AMP-activated protein kinase (AMPK), Neuropeptide Y receptor Y2 (NPY2R), leptin (LEP), ghrelin (GHRL), proopiomelanocortin (POMC), and adiponectin (ADIPOQ) were quantified using RT-qPCR. Body composition was assessed via skinfold measurements and compared and correlated between cohorts. Feeding outcomes were recorded. RESULTS/ANTICIPATED RESULTS: 23 infants were recruited in each cohort. POMC and AMPK were expressed by 71% and 88% of infants respectively in both cohorts. NPY2R was expressed by 79% and 83% of the diabetic cohort and normoglycemia cohort respectively, while GHRL was expressed by 75% and 79% of the diabetic cohort and normoglycemia cohort, respectively. LEP and ADIPOQ were not reliably expressed in either cohort. Infants with a higher body fat percentage were less likely to express NPY2R (OR= 0.76). There was no significant association between body fat percentage and expression of AMPK, POMC, or GHRL. Only 3 IDMs were noted by providers to exhibit poor oral intake, limiting our ability to correlate gene expression and body composition with feeding outcomes. DISCUSSION/SIGNIFICANCE: Non-invasive assessment of hunger signaling gene expression is possible through salivary analysis of AMPK, POMC, NPY2R, and GHRL. Given the paucity of IDMs with poor feeding in our study, future studies should target IDMs requiring feeding support to understand mechanisms driving aberrant feeding behavior.
235 Analysiss of TNBC Cell Lines Cultured a Novel Translational Breast Cancer Microphysiological System (BC-MPS)
Authors: Hebert Katherine L. , Nguyen Khoa , Cheng Thomas , Alzoubi Madlin , Elliott Steven , Collins-Burow Bridgette , Martin Elizabeth , Lau Frank , Burow Matthew
Published: April 2022

OBJECTIVES/GOALS: Current approaches to drug development for the aggressive triple negative breast cancer rely on current 2D and 3D in vitro models which have limited capabilities. We have developed a translational microphysiological system that can maintain the human breast microenvironment to capture the complex interaction with the tumor microenvironment. METHODS/STUDY POPULATION: Three different TNBC cell lines were seeded in BC-MPS: MDA-MB-231 parental cell line, MDA-MB-231wiht the gene, LKB1 overexpressed, which is a tumor suppressor, and MDA-MB-231 with the enzyme, ERK5, an enzyme associated with increased metastasis and drug resistance, knocked out. These three TNBC cell lines were cultured in a standard 2D 96-well plate and in BC-MPS. Time-lapse videos were taken to track cellular mobility. RNA-sequencing was performed to compare different expression levels of various cancer related genes of the cell lines cultured in standard 2D and BC-MPS. RESULTS/ANTICIPATED RESULTS: The LKB1 overexpressed MDA-MB-231 and the ERK5-ko MDA-MB-231 cell lines are expected to have decreased mobility compared to the parental cells. The cell lines are expected to have increased expression of cancer related genes when cultured in BC-MPS than when cultured in standard 2D due to the presence of human breast tissue. DISCUSSION/SIGNIFICANCE: BC-MPS is a promising new translational MPS that facilitates studying long term interactions between real human breast tissue and cancer cells. The BC-MPS systems ability to support the growth of established cell lines has been demonstrated. Future studies will focus on developing the model for personalized medicine.
241 Utilizing real-world evidence to increase efficiency of randomized controlled trials with application to repurposed therapeutics for COVID-19
Authors: Haine Lillian M F , Murray Thomas A , Koopmeiners Joseph S
Published: April 2022

OBJECTIVES/GOALS: We aim to extend a novel statistical method called the Semi-Supervised Mixture Multisource Exchangeability Model (SS-MIX-MEM) and to implement the SS-MIX-MEM approach to supplement ALPS-COVID data with N3C data to achieve analyses with greater precision and actionable conclusions. METHODS/STUDY POPULATION: We will apply the SS-MIX-MEM to supplement the Angiotensin receptor blocker-based Lung Protective Strategy for COVID-19 (ALPS-COVID) RCTs with the National COVID Cohort Collaborative (N3C) database. ALPS-COVID includes both an inpatient and outpatient trial, which investigate losartan as a treatment for COVID-19. The outpatient trial sought to randomize 580 individuals but only enrolled 117, whereas the inpatient trial met its enrollment target and randomized 205 individuals. The N3C database has 3,237,344 COVID-19 cases alongside demographics, lab values, and more. RESULTS/ANTICIPATED RESULTS: In simulation studies, the proposed SS-MIX-MEM approach effectively leveraged a subgroup of supplemental real world data for RCT analyses, improving trial efficiency by increasing precision of treatment effect estimates, decreasing necessary sample size, and introducing minimal bias. In an influenza trial real world data application, the SS-MIX-MEM approach was able to effectively provide insight into treatment effect heterogeneity found in an RCT analogous to incorporating around 80 individuals into a subgroup analysis. We anticipate that leveraging external real world data in a re-analysis of the ALPS-COVID RCTs could provide new insights into losartan, a readily available, potentially beneficial therapeutic for COVID-19. DISCUSSION/SIGNIFICANCE: The high blood pressure drug, losartan, is readily available, has an established safety profile, and might be effective as a treatment for COVID-19. Given that we have very few effective treatment options and are still in the midst of a global pandemic, patients with COVID-19 would greatly benefit from a repurposed, readily available treatment.
244 A Systematic Review of Epileptiform Changes During Sevoflurane Anesthesia In Infants and Children
Authors: Gizzo Luciana , Ivenitsky Daria , Ferrera Alexander , Tam Matthew , Legatt Alan D. , Yozawitz Elissa G. , Lo Yungtai , Li Guohua , Shinnar Shlomo , Chao Jerry
Published: April 2022

OBJECTIVES/GOALS: Early clinical case reports have described incidental epileptiform changes during electrophysiological monitoring. The objective of this study was to perform a systematic review of all existing investigations of epileptiform activity during sevoflurane use in pediatric anesthesia. The heterogenous EEG data will be analyzed in a meta analysis METHODS/STUDY POPULATION: A targeted, PICO-based clinical question was crafted and registered a priori on PROSPERO on 3/19/21. Under the guidance of a librarian from the Albert Einstein College of Medicine, a boolean search string was generated to search articles and gray literature for terms such as pediatric, sevoflurane and electroencephalogram in PubMed, OVID, Cochrane, Google Scholar, etc. We utilized the software platform tool COVIDENCE to manage our review. 495 references were imported for initial screening. 56 English-language, full-text studies were included for further review. The final 13 references were included in data extraction and Newcastle-Ottawa bias assessment. The characteristics of the studies and their primary outcomes were collected in tabular form. Strategies for data synthesis were discussed weekly. RESULTS/ANTICIPATED RESULTS: Epileptiform changes reported in the literature during pediatric sevoflurane anesthesia ranged from 0 - 95%. EEG data were acquired using a variety of recording systems with variable number of leads and heterogeneous outcomes reported. The periods of anesthesia monitoring were also heterogeneous. Characteristics of the studies are presented in Table 1. 495 references were imported for screening with 13 final references for data extraction. EEG abnormalities were reported in 204/649 (31.4%) subjects ranging in age from neonate to 18 years; the majority of studies utilized less than 16 channels of (10/13, 76.9%) (Table 1). There was variability in sevoflurane dosing, premedication (e.g., midzolam, hydroxyzine), and periods of anesthesia monitored. DISCUSSION/SIGNIFICANCE: There was heterogeneity noted across reviewed literature including study design, phases of anesthesia, ventilation methods, number of EEG leads recorded and adjuvant anesthetics administered. Nevertheless, this review rigorously classified epileptiform activity during Sevoflurane thereby influencing modern anesthesia.
250 Defining Developmentally High-Risk Full Term and Late Preterm Infants in the Neonatal Intensive Care Unit
Authors: Carlton Katherine , Cabacungan Erwin , Cohen Susan
Published: April 2022

OBJECTIVES/GOALS: We aim to describe the preschool age developmental outcomes of children born full term or late preterm requiring care in the Childrens Wisconsin (CW) neonatal intensive care unit (NICU). Our objective is to develop a model to predict which NICU infants are at high risk for abnormal preschool age development based on their neonatal characteristics. METHODS/STUDY POPULATION: Retrospective cohort study including full term (â‰¥ 37 weeks) and late preterm (34-36 weeks) infants admitted to the CW NICU between 1/1/2013-12/31/2015 with a developmental assessment by a general practitioner or neuropsychology evaluation between 3-6 years of age. Data extraction will include maternal history, neonatal and delivery factors, and developmental evaluation results. Descriptive statistics will be used to determine the proportion of patients with abnormal preschool development stratified by NICU diagnoses, surgical procedures, subspecialty involvement, and patient demographics. Decision tree analysis will be performed to generate a prediction model identifying those infants at higher risk than their peers for an abnormal developmental assessment at 3-6 years of age. RESULTS/ANTICIPATED RESULTS: Analysis to date reveals 1360 patients â‰¥ 34 weeks gestation admitted to the CW NICU between 1/1/2013-12/31/2015. 80 patients received neuropsychology evaluations and the remaining 1280 patients had general practitioner developmental assessments. We anticipate that those infants referred for neuropsychology evaluation will have a higher proportion of abnormal developmental assessments when compared to those patients with routine general practitioner assessments. In addition, we hypothesize that common neonatal factors among the infants who received neuropsychology assessments will best predict abnormal preschool development within the decision tree analysis. DISCUSSION/SIGNIFICANCE: This study is one of the first to define risk factors in full term and late preterm infants cared for in the NICU associated with abnormal preschool age development. Identifying these infants at high-risk early on will allow providers to initiate early developmental interventions, schedule close NICU follow-up care, and provide parental counseling.
270 Neonipple Formation After Implantation Of Acellular Ovine Xenograft
Authors: Vernice Nicholas Andrew , Caughey Sarah , Berri Nabih , Dong Xue , Harris Jason , Bender Ryan J. , Spector Jason A.
Published: April 2022

OBJECTIVES/GOALS: To determine if decellularized costal cartilage (DCC), which could theoretically be obtained “off the shelf,” would provide similar results to autologous cartilage grafts previously studied in this lab, thereby widening the application of our novel nipple engineering approach to all patients undergoing nipple reconstruction. METHODS/STUDY POPULATION: PLA scaffolds (diameter: 1.0 cm, height: 1.0 cm) were printed using a PRUSA 3D printer and sterilized. Lamb costal cartilage was minced (1 mm3) or zested (<0.2 mm3) and then decellularized. The quality of decellularization was assessed using DNA quantification and histological analysis. DCC was then packed into PLA scaffolds and implanted subcutaneously into immunocompetent Sprague Dawley rats using a CV flap technique. The constructs were explanted and evaluated up to 6 months after implantation. RESULTS/ANTICIPATED RESULTS: All nipple reconstructions showed well-preserved diameter and projection due to persistence of the external scaffolds at 1, 3, and 6 months. Mass and volume of engineered tissue was well-preserved over all timepoints. Compared to implantation values, engineered zested nipples demonstrated a 12% mass increase and a 22% volume increase at 6 months. Minced nipples illustrated a similar mass and volume gain with a 21% increase in mass and a 13% increase in volume at 6 months secondary to infiltration of fibrovascular tissue and growth through scaffold wall pores, respectively. Histologic analysis demonstrated a mild inflammatory infiltrate 1 month after implantation which was replaced by fibrovascular tissue by 3 months that remained stable through 6 months. The processed DCC structure remained unchanged over time. DISCUSSION/SIGNIFICANCE: Using acellular ovine xenograft within bioabsorbable scaffolds, we have engineered neonipples that maintain their volume for at least 6 months. DCC architecture is well-preserved with minimal evidence of immune-mediated degradation. By using DCC, this novel approach to nipple engineering may be applied to any patient requiring reconstruction.
281 A Knowledge Chasm: Preoperative Mental Illness Impact on New Onset Postoperative Atrial Fibrillation
Authors: Kolba Natalie , Shroyer Anne Laurie W. , Bilfinger Thomas
Published: April 2022

OBJECTIVES/GOALS: To date, no reviews have been published combining the topics of pre-operative mental illness and postoperative atrial fibrillation (POAF). For adult cardiac surgical procedures, therefore, this scoping review provides an overview of the published evidence for this novel topic. METHODS/STUDY POPULATION: A combination of MESH terms and text words were used to capture all papers analyzing any risk factors for atrial fibrillation development after any cardiovascular surgery, and thus, 4,203 search results were initially identified. Papers were manually screened for relevance, resulting in 69 articles that analyzed risk factors associated with POAF. As a separate analytical step, all included publications were used to identify additional citations and Web of Science PubMed identification number-based (i.e., PMID) specific backwards citation searches were performed to ensure comprehensive coverage. For each of the three articles analyzed, moreover, these PMIDs were further evaluated using individual google scholar searches to assure no other relevant literature was inadvertently found. RESULTS/ANTICIPATED RESULTS: For the 69 relevant articles found with multivariable risk models predicting new onset POAF during this scoping review, a wide variety of patient risk factors were reported. The statistically significant predictors reported for new POAF included age (57 papers), hypertension (22 papers), tobacco use (15 papers), LVEF-related characteristics (14 papers), COPD (13 papers), and patient gender (13 papers). All other factors were found to be significant 10 times or fewer; importantly, no multivariable models evaluated mental illness-related factors as predictors of new onset POAF. Only 3 articles even considered the potential influence of mental illnesses. DISCUSSION/SIGNIFICANCE: Mentally ill patients have been documented to have increased incidence of cardiovascular disease and adverse cardiac outcomes, however, very few articles evaluated mental illness associations with POAF. This comprehensive review has demonstrated the urgent need for new research focused on mental illness impacts.
282 Follow-up for Evaluation for Nerve Repair Surgery: The Role of Socioeconomic and Geographic Factors in Victims of Violent Nerve Injury
Authors: Liebendorfer Adam , Marsh Evan , Finnan Michael , Bruce Jordan , Skolnick Gary , Kells Amy
Published: April 2022

OBJECTIVES/GOALS: We have created the first-ever database for all patients who have acute peripheral nerve injuries. Our goal is to use this data to construct two clinical scoring systems to facilitate triaging by emergency room physicians by 1.) identifying patients who would likely benefit from a nerve surgery, and 2.) identify patients who are less likely to follow up. METHODS/STUDY POPULATION: Based on 2,606 consult emails from the Washington University in St. Louiss Division of Plastic and Reconstructive Surgery, from 2013 to 2019, we have identified more than 600 patient cases in which patients presented after an injury with any kind of nerve deficit. We have defined nerve deficits as any abnormality on motor or sensory exam of the extremities, excluding those younger than 18 years old and those with other disease processes that may affect function of the peripheral nervous system. Data for these patients was collected from electronic medical records and include handedness, nerve injured, operative details, area deprivation index decile, mechanism of injury, insurance and employment status, and whether injuries were a result of work or violence. RESULTS/ANTICIPATED RESULTS: Our plan is to provide the first descriptive statistics for such a population, as well as multiple regression analysis for our two endpoints, loss to follow up and need for surgery. Following regression analysis, we also look to perform cluster analysis to further study co-occurring clinical and social features that affect our endpoints. We are excitedly awaiting to perform the analysis after the final data has been collected. DISCUSSION/SIGNIFICANCE: Nerve injury is a common symptom of penetrating injuries. Many of the socioeconomic factors associated with increased risk for violent injury are also risk factors for decreased rates of follow up. This problem is particularly worrisome given that nerve repair surgery is only viable as an option to return function within the first year after injury.
283 The Impact of Minimum Volume Thresholds on Geographic Access to Stroke Thrombectomy
Authors: Leykina Liza , Guterman Elan , Hsia Renee , Kim Anthony
Published: April 2022

OBJECTIVES/GOALS: At least 15 stroke thrombectomies per proceduralist per year are required for Thrombectomy-Capable and Comprehensive Stroke Centers. We sought to estimate the potential impact of these minimum volume thresholds on geographic access to stroke thrombectomy (ST). METHODS/STUDY POPULATION: Using the Florida State Inpatient Database, we will identify patients discharged with acute ischemic stroke from 2015-2019 and all non-federal facilities that performed ?1 ST procedure per year. We will then calculate the proportion of stroke patients who live within 20, 65, 115, 165 and 200 miles (correlating with estimated ground transport times of 1, 2, 3, 4 and 5 hours, respectively) of centers that perform ST using ArcGIS software and evaluate the impact of varying the threshold ST volume required by each facility on this proportion. We will then perform multiple two proportion z-tests to compare proportions of patients within driving distance over time. RESULTS/ANTICIPATED RESULTS: We hypothesize that over time, and particularly after the pivotal trials of ST were published in 2015, that the number of facilities that perform ST have increased over time, which would increase the geographic access to ST. We also hypothesize that since the Joint Commission set the minimum procedural volume for proceduralists at Thrombectomy-Capable and Comprehensive Stroke Center to 15 per year, this would work to increase regionalization and could work to decrease geographic access to ST. However, we hope to elucidate the net impact of the interplay between these two opposing factors on regionalization of care over time which is currently unclear. DISCUSSION/SIGNIFICANCE: Current ST volume thresholds have focused on technical proficiency but may impact regionalization of care and geographic access to ST. Since access to ST is time-sensitive, a data-driven approach and better coordination on a regional level may be necessary to ensure timely access to ST.
300 Improving the diagnosis and classification of facial pain conditions with MRI-based features
Authors: Mulford Kellen , Moen Sean , Grande Andrew W. , Nixdorf Donald R. , Van de Moortele Pierre-Francois
Published: April 2022

OBJECTIVES/GOALS: Trigeminal Neuralgia (TN) is a debilitating neuropathic condition characterized by electric-shock-like pain attacks. TN is considered a clinical diagnosis, and few proposed objective markers exist. This work studies the ability of advanced MRI techniques to diagnose and classify TN. METHODS/STUDY POPULATION: Anatomical MRI data from patients undergoing radiosurgery to treat TN was collected. A custom deep-learning UNet algorithm was trained to segment trigeminal nerves from the pons to the anterior wall of Meckels cave using segments drawn by an expert in neuroanatomy. 108 radiomics features related to nerve shape, voxel intensity, and image texture were extracted from the segmented nerves. A 2 layer neural network was trained to distinguish TN affected nerves from the pain-free contralateral nerves. Feature selection was performed within a cross-validation scheme to prevent model overfitting. Mean model performance over the validation sets was used to estimate model generalizability. RESULTS/ANTICIPATED RESULTS: 134 patients and 268 nerves were included. The average number of years with TN was 8. The average validation set accuracy was 78% [range: 75-80%]. The average validation set sensitivity and specificity were 0.82 [range: 0.79-0.84] and 0.76 [range: 0.70-0.79]. 34% of patients had undergone a prior invasive procedure to treat their TN. To evaluate whether the model detected signal changes relating to the previous treatment, those patients were excluded and the model was retrained on the surgically naive patients. Model performance in a reduced cohort of patients was similar to the model trained on all the patients, with accuracy of 77% [range: 73-82%]. DISCUSSION/SIGNIFICANCE: This study suggests that radiomics features calculated from MRIs of trigeminal nerves correlate with anatomical changes in TN affected nerves. This technique will need to be verified in a larger, more heterogeneous cohort of TN patients with a range of MRI acquisition parameters.
318 Building A Better CAR: Improving CAR-T Trafficking in Cancer Therapy
Authors: Hong Yeonsun , Kim Hye-Ran , Walling Brandon L. , Lozada John , Amitrano Andrea M. , Sailer Cooper J. , Lim Kihong , Mongre Raj Kumar , Kim Kyun-Do , Capece Tara , Lomakina Elena B. , Reilly Nicholas S. , Waugh Richard E. , Reagan Patrick M. , Kim Minsoo , Oakes Patrick W. , Jun Chang-Duk
Published: April 2022

OBJECTIVES/GOALS: #NAME? METHODS/STUDY POPULATION: Cell culture & protein identification: human T cells were purified from healthy blood, then activated & cultured for 5d. CAR-T cells were collected from infusion bags of cancer patients undergoing CAR-T. Silver staining of naive & activated healthy T-cell lysates was compared; B-II spectrin was upregulated and confirmed by Western blot. Migration assays: naive & activated T-cells were imaged during migration on ICAM-1 and ICAM-1 + CXCL12 coated plates. T-cells were transfected with BII-spectrin cDNA & the chemokine dependence of migration was compared with controls. In-vivo studies: in a melanoma mouse model, BII-spectrin transfected or control T-cells were injected; tumors were followed with serial imaging. Human patient records were examined to correlate endogenous BII-spectrin levels and CAR-T response. RESULTS/ANTICIPATED RESULTS: Activated T-cells downregulate the cytoskeletal protein B-II spectrin compared to naive cells, leading to chemokine-independent migration in in vitro assays and off-target trafficking when CAR-T cells are given in vivo. Restoration of B-II spectrin levels via transfection restores chemokine-dependence of activated T-cells. In a mouse melanoma model, control mice injected with standard activated T-cells showed fewer cells in the tumor site and more cells in the off-target organs (spleen, lungs) when compared to mice injected with B-II spectrin transfected cells. Furthermore, among 3 human patients undergoing CAR-T therapy, those with higher endogenous B-II spectrin levels experienced fewer side-effects, measured by the neurotoxicity and cytokine release syndrome grades. DISCUSSION/SIGNIFICANCE: A major hurdle to widespread CAR-T therapy for cancer is significant, often fatal side-effects. Our work shows that the protein B-II spectrin is downregulated during CAR-T production, and that restoring B-II spectrin levels decreases side-effects while increasing tumor clearance--hopefully translating to better CAR-T regimens for the future.
321 Impact of CDK4/6 inhibitor-induced cellular senescence on stromal responses to breast cancer
Authors: Gallanis Gregory T. , Wellstein Anton , Riegel Anna T.
Published: April 2022

OBJECTIVES/GOALS: The objective of my study is to test my overall hypothesis that CDK4/6 inhibitor-induced cellular senescence has long-lasting consequences on the tumor microenvironment that affect tumor growth and disease progression. METHODS/STUDY POPULATION: There is a lack of mechanistic understanding of the crosstalk of drug effects on different cellular pathways. One less-studied consequence of CDK4/6 inhibitors (CDKI) is their ability to induce cellular senescence in the non-malignant host cells (“stroma”), which causes secretion of a plethora of proteins that have lasting effects on the tumor microenvironment. Stromal cells are known to be able to enter senescence and promote a state of chronic inflammation that contributes to disease progression. I propose to study the stromal contribution to the tumor microenvironment and how this affects tumor growth and disease progression by using genetically engineered mouse models of breast cancer. RESULTS/ANTICIPATED RESULTS: We have found that palbociclib, a CDKI, inhibited growth and induced senescence of breast cancer cell lines xenografted into mice, and that while most on-target effects of palbociclib were reversed after treatment cessation, there was long-lasting down-regulation of interferon-Î³ and inflammatory response signaling pathways in the mouse host stroma that persisted after cessation of palbociclib treatment. In syngeneic models, we have found that palbociclib treatment influenced migration of T cells into CDKI-resistant tumors. DISCUSSION/SIGNIFICANCE: These studies will shed light on how senescence-inducing anticancer therapies, such as CDKI, affect tumor growth and disease progression. My studies will help elucidate new therapeutic avenues in combination with CDKI to combat therapeutic resistance in advanced breast cancer.
329 Longitudinal immune profiling reveals unique myeloid and T cell phenotypes associated with spontaneous immunoediting in a novel prostate tumor model
Authors: Ager Casey , Obradovic Aleksandar , Arriaga Juan M. , Chaimowitz Matthew G. , Abate-Shen Cory , Califano Andrea , Drake Charles G.
Published: April 2022

OBJECTIVES/GOALS: Few preclinical models exist to study how tumors transition from prolonged stable disease ('equilibrium') to progressive disease ('escape'). We characterized a new murine tumor model the exhibits such behavior, and sought to identify and validate the role of unique tumor-infiltrating immune cell subsets in this process. METHODS/STUDY POPULATION: We evaluated growth of NPK-C1 (originally LM7304; received from Dr. Cory Abate-Shen at Columbia University), a cell line developed from spontaneous prostate cancer lung metastases in NPK mice (Nkx3.1CreERT2/+Ptenflox/floxKrasLSL-G12D/+R26R-LSL-YFP/+), in immune competent (C57BL/6) and immune deficient mice (J/Nu). We determined the role of CD4 and CD8 T cells in regulating the 'equilibrium to escape’ growth dynamics of NPK-C1 via in vivo cell depletions at key inflection points of tumor growth. To deeply profile the immune contexture of NPK-C1 at these inflection points, we developed a 28-color immunophenotyping panel for use on a Cytek Aurora spectral flow cytometer. We performed dimensionality reduction and clustering analyses on these data using tSNE and FlowSOM algorithms within FlowJo (v10.6). RESULTS/ANTICIPATED RESULTS: We found that activated CD4 effector T cells are enriched in regressing NPK-C1 tumors, highlighting a role for CD4 T cells in antitumor immunity. CD8 T cells are also important for NPK-C1 control; specifically central memory-like cytotoxic CD8 T cells. Depletion of either CD4 or CD8 T cells during the equilibrium phase of NPK-C1 growth confirmed the role of these cells in antagonizing NPK-C1 escape. Tregs as a whole were counterintuitively enriched in regressing tumors, however high dimensional analysis reveals their significant phenotypic diversity, with a number of Treg subpopulations enriched in progressing tumors. In the myeloid compartment, we found that iNOS+ DC-like cells are enriched in regressing tumors, while CD103+ DCs are associated with late stage tumor progression. DISCUSSION/SIGNIFICANCE: In total, these analyses of the NPK-C1 model provide novel insights into the roles of lymphoid and myeloid populations throughout key phases of tumor/immune co-evolution, and highlight a role for multi-dimensional flow cytometry-based analyses to more deeply understand immune cell dynamics in the tumor microenvironment.
345 Interest in and Perceived Effectiveness of Contingency Management Among Alcohol Drinkers Using Behavioral Economic Purchase Tasks
Authors: Traxler Haily , Traxler Haily , Kaplan Brent , Rzeszutek Mark , Koffarnus Mikhail
Published: April 2022

OBJECTIVES/GOALS: The purpose of this study was to develop behavioral economic purchase tasks to assess interest in CM as a function of treatment cost and perceived effectiveness of CM as a function of abstinence incentive size in alcohol drinkers. Additionally, these purchase tasks are being assessed for their clinical utility in an ongoing clinical trial. METHODS/STUDY POPULATION: Alcohol drinkers recruited from Amazon Mechanical Turk completed behavioral economic purchase tasks measuring demand for CM based on targeted abstinence intervals and treatment effectiveness and alcohol use disorder severity assessments. Nonlinear mixed effects modeling was used to fit demand curves and assess the relationship between individual characteristics and demand metrics for CM. Ongoing analyses involve administering the same behavioral economic purchase tasks in heavy alcohol users in the ongoing clinical trial, which is aimed at reducing alcohol use through remotely implemented CM. RESULTS/ANTICIPATED RESULTS: Mechanical Turk participants reported higher probability of abstinence when offered larger incentives and required larger incentives when duration of abstinence required to earn the incentive was increased. Additionally, willingness to pay for treatment increased as effectiveness of treatment increased. It is anticipated that these patterns will be observed in the clinical trial participants. DISCUSSION/SIGNIFICANCE: Abstinence interval and treatment effectiveness are important features to consider when developing effective CM for widespread use, as these variables affected participants likelihood of being abstinent and their interest in treatment. We are currently working on verifying the results of these assessments in clinical trial participants.
353 MFM Provider Adherence to USPSTF Low Dose Aspirin Guidelines for Preeclampsia Prevention in Nulliparous Patients
Authors: Myers Sabrena O. , Myers Sabrena O. , Luke Shauntell , Ramey-Collier Khaila , Truong Tracy , Weaver Kristin , Swamy Geeta , Wheeler Sarahn
Published: April 2022

OBJECTIVES/GOALS: Recent studies suggest nulliparous patients benefit from low dose aspirin (LDA), yet there are limited studies examining MFM providers adherence to USPSTF guidelines and predictors of adherence. We identified demographic, obstetric, and clinical characteristics associated with guideline concordant counseling on LDA in nulliparous women. METHODS/STUDY POPULATION: Retrospective cohort study of pregnant nulliparous patients who received MFM prenatal care at a single tertiary center (1/1/2019-6/30/2020). Multiple gestations, > 2 spontaneous or therapeutic abortions, and contraindications to LDA were excluded. Maternal demographic and clinical characteristics were abstracted from the electronic medical record. The primary outcome was documented LDA counseling based on USPSTF guidelines. Data were analyzed using bivariate analysis and logistic regression using R 4.1.0 (R Core Team, 2021). RESULTS/ANTICIPATED RESULTS: Among 394 records in the analysis cohort, 316 met USPSTF guidelines for LDA. 164 (51.9%) meeting guidelines had documented LDA counseling. 67.4% of individuals with ?1 major USPSTF risk factors were counseled and 50.7% with ?2 moderate risk factors were counseled (Table 1). Age at the estimated due date (EDD), Black or Other race, chronic hypertension, and obesity are significantly associated with higher odds of aspirin counseling (Table 2). Patients with chronic hypertension had 4.15 higher odds of receiving low dose aspirin counseling compared to non-hypertensive patient (Table 2). DISCUSSION/SIGNIFICANCE: Our results suggest that only 51.9% of patients eligible for LDA received counseling despite MFM care. Increasing MFM provider awareness about the USPSTF guidelines and creating tools that facilitate guideline concordant counseling may increase the number of eligible patients who are counseled about LDA.
364 Identification of Symptom-Based Phenotypes in PASC Patients through Bipartite Network Analysis: Implications for Patient Triage and Precision Treatment Strategies
Authors: Bhavnani Suresh K. , Zhang Weibin , Hatch Sandra , Urban Randall , Tignanelli Christopher
Published: April 2022

OBJECTIVES/GOALS: Approximately 10% of COVID-19 patients experience multiple symptoms weeks and months after the acute phase of infection. Our goal was to use advanced machine learning methods to identify PASC phenotypes based on their symptom profiles, and their association with critical adverse outcomes, with the goal of designing future targeted interventions. METHODS/STUDY POPULATION: Data. All COVID-19 outpatients from 12 University of Minnesota hospitals and 60 clinics. Independent variables consisted of 20 CDC-defined PASC symptoms extracted from clinical notes using NLP. Covariates included demographics, and outcomes included New Psychological Diagnostic Evaluation, and Number of PASC Hospital Visits (>=5). Cases (n=3235) consisted of patients with at least one symptom, and controls (n=3034) consisted of patients with no symptoms. Method. (1) Used bipartite network analysis and modularity maximization to identify patient-symptom biclusters. (2) Used multivariable logistic regression (adjusted for demographics and corrected through Bonferroni) to measure the odds ratio of each patient bicluster to adverse outcomes, compared to controls, and to each of the other biclusters. RESULTS/ANTICIPATED RESULTS: The analysis identified 6 PASC phenotypes (http://www.skbhavnani.com/DIVA/Images/Fig-1-PASC-Network.jpg), which was statistically significant compared to 1000 random permutations of the data (PASC=.31, Random Median=.27, z=11, P<.01). Three of the clusters (Cluster-1, Cluster-4, and Cluster-5 encircled with ovals in Fig. 1) contained CNS-related symptoms, which had statistically significant risk for one or both of the adverse outcomes. For example, Cluster-1 with critical CNS symptoms (depression, insomnia, anxiety, brain-fog/difficulty-thinking), had a significantly higher OR compared to the controls for New Psychological Diagnostic Evaluation (OR=6.6, CI=4.9-9.1, P-corr<.001), in addition to having a significantly higher ORs for the same outcome compared to all the other clusters. DISCUSSION/SIGNIFICANCE: The results identified distinct PASC phenotypes based on symptom profiles, with three of them related to CNS symptoms, each of which had significantly higher risk for specific adverse outcomes compared to controls. We will test whether these phenotypes replicate in the N3C data, and explore their translation into triage and treatment strategies.
392 Adapting a randomized, placebo-controlled pilot study aimed to reduce anxiety symptoms to overcome recruitment barriers
Authors: Smith Kristen S , Frugé Drew
Published: April 2022

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.
395 Vascular Cognitive Impairment: Novel Endothelial Mechanisms and the Impact of Dietary PUFAs
Authors: Tomberlin Jensen , Albayram Onder , Kurtz John , Karakaya Eda , Ergul Adviye
Published: April 2022

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.
403 Can supervision support implementation of evidence-based practices in substance use disorder treatment programs? A qualitative analysis of organizational and environmental contexts in Arkansas
Authors: Baloh Jure , Curran Geoffrey M.
Published: April 2022

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.
405 Role of Glycans in Cancer Associated Fibroblast-Derived Exosome Immunoregulation
Authors: Anderson Johnathon D. , Nolta Jan , Farwell D. Gregory , Birkland Andrew , Van de Water Judy , Carraway Kermit , Lebrilla Carlito
Published: April 2022

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.
410 Mild Maternal Undernutrition Results in a Premature Neonatal Leptin Surge and Resistance in Male Offspring to a High Fat Diet
Authors: Miles Tiffany K. , Allensworth Melody L. , Silva Moreira Ana Rita , Odle Angela K. , Haney Anessa , Lagasse Alexandra , MacNicol Angus M. , Childs Gwen V.
Published: April 2022

OBJECTIVES/GOALS: Maternal undernutrition, a form of malnutrition, can alter neonatal leptin signaling and result in metabolic dysfunction in adulthood. We developed a mild undernutrition model to relate more to societys nutritional challenges and to test the hypothesis that a shift in the neonatal leptin surge would result in sex-specific metabolic changes. METHODS/STUDY POPULATION: We studied pups from undernourished dams which were calorically restricted by 20% (CR20) from embryonic day 15 until postnatal day (PND) 21. We tested 216 offspring from 11 Fed dams and 13 undernourished dams (CR20), detecting a leptin surge in control fed progeny at PND11. At 3 months of age, offspring from 3 dams per maternal nutrient status were either exposed to a 45% high fat diet (HFD) or control diet (10% fat) for 16 weeks. Anterior pituitary hormones were analyzed in the pituitary and serum of neonates and adults. To determine the mechanism of the phenotype observed in male adult offspring on the HFD, single cell RNA sequencing was used to analyze the pituitary, fat and liver. RESULTS/ANTICIPATED RESULTS: Offspring of CR20 dams had an early leptin surge peaking at PND8 and GH levels at PND1 were higher in CR20 progeny. Weights of both male and female CR20 offspring were lower and body lengths were shorter than controls. As adults, Fed mice from both sexes had increased weight gain with HFD. However, although CR20 females gained weight on the HFD, male progeny from CR20 dams did not gain weight on the HFD and appeared protected from impact. We found sex-specific changes in pituitary Gh, Ghrhr, and Ghsr mRNA levels. Single cell RNA sequencing of pituitary, fat and liver of male offspring showed significant regulation of transcripts in fat of male offspring from Fed dams that was not found in CR20 males when compared to control fed mice. DISCUSSION/SIGNIFICANCE: Mild undernutrition causes a prematurely high leptin surge and sex-specific growth responses to a HFD, including resistance to a HFD in underfed males. Transcript analysis in fat of males resistant to HFD induced obesity may reveal mechanisms that provide protection against HFD induced weight gain.