bluuebunny/crossref_metadata_2025 huggingface dataset:
294 Identification of MCAK Inhibitors that Induce Aneuploidy in Triple Negative Breast Cancer Models
Authors: Smith John , Husted Stefan , Pilrose Jay , Kuchangi Disha , Ems-McClung Stephanie C. , Carpenter Richard L. , Walczak Claire E.
Published: April 2023

OBJECTIVES/GOALS: Microtubule poisons, like Taxol, are used to treat triple negative breast cancer (TNBC) and may induce lethal aneuploidy in cancer cells. Patients initially respond, but often develop drug resistance. New targeted drugs that cause aneuploidy may be a valuable approach to therapy. One potential target is the Kinesin 13 MCAK, which limits aneuploidy. METHODS/STUDY POPULATION: TCGA and GSE47561 databases were probed for MCAK expression, and data was stratified by subtype and survival statistics. Knockdown studies were performed to test whether MCAK knockdown sensitizes cells to taxanes for cell proliferation and for induction of aneuploidy. FRET and image-based screens were used to identify MCAK inhibitors from small molecule inhibitor libraries. Inhibitors were then tested for functional effects in multiple cell-based assays and for clonal growth in colony formation assays. RESULTS/ANTICIPATED RESULTS: MCAK expression is upregulated in TNBC and associated with reduced overall survival. Knockdown of MCAK caused a two-to-five-fold reduction of the IC50 for Taxol in cancer cell lines, with no change in normal cell lines. Taxol treatment or MCAK knockdown increased aneuploidy induction, with no additive effect between the two. Our small molecule screen identified three putative MCAK inhibitors, which induced aneuploidy in both taxane-sensitive and taxane-resistant cells. These inhibitors also reduced clonogenic growth, and the most potent inhibitor, C4, caused an approximate five-fold reduction in the IC50 for Taxol in cell proliferation assays. DISCUSSION/SIGNIFICANCE: MCAK can serve as a biomarker of breast cancer prognosis. MCAK knockdown or inhibition sensitizes cancer cells to Taxol without affecting normal cells, making it a potential target in combination therapy. MCAK inhibitors also reduce growth as single agents in taxane resistant lines, giving them potential use as therapies in resistant disease.
304 Let Kids Play: Using Virtual Reality as a Substitute for General Anesthesia for Minor Procedures in Children
Authors: Chao Stephanie , Jazi Fereshteh Salimi , Sabapaty Akanksha , Rafeeqi Talha , Dalusag Kyla Santos
Published: April 2023

OBJECTIVES/GOALS: Minor procedures are anxiety-provoking and/or painful for children. Virtual reality (VR) is an emerging technology in medicine and largely has been used as an adjunct to analgesia and opioids. This study reviews the institutional use of VR in lieu of pharmacology and general anesthesia (GA) to perform minor procedures in a pediatric population. METHODS/STUDY POPULATION: A retrospective chart review was performed on all patients that presented to our institution from 2019 to 2022 for hormone implant placement, exchange, or removal with VR distraction. Demographic and procedure information was recorded. The primary outcome was successful procedure completion without requiring pharmacologic sedation or analgesia. RESULTS/ANTICIPATED RESULTS: A total of 111 patients underwent the following minor procedures with VR only. 14 patients underwent more than one procedure resulting in a total of 126 procedures. The mean age was 11.3 ± 3.6 years. 43 patients were female, 23 were female to male, 6 were non-binary, 7 were male, and 32 were male to female. 58% had private insurance. The most common diagnosis was precocious puberty (54%) followed by gender dysphoria (46%). The most common procedure was implant placement (72%). 69% of procedures were performed in the clinic and 31% in a procedural room. All procedures were completed without requiring sedation or GA. None of the patients required intravenous catheter placement for the procedure. No intra-procedural complications were recorded. DISCUSSION/SIGNIFICANCE: Despite the current trend toward minimizing GA and sedation in children, there is no widely accepted substitute. VR is a feasible option that can spare children from sedation or GA for minor procedures. This can enable procedures to be transitioned into more resource efficient settings and improve pediatric patient experience.
317 Omics based analysis to identify novel markers of TMZ response in Glioblastoma Multiforme
Authors: Reed Megan R. , De Loose Annick , Guzman Grace , Lyle A. Geoffrey , Learned Katrina , Vaske Olena M. , Rodriguez Analiz
Published: April 2023

OBJECTIVES/GOALS: MGMT methylation status is used to predict the response to TMZ. However, a subpopulation of patients lacking MGMT methylation still respond to TMZ. We applied omics approaches and functional studies to a cohort of GBM patients to identify novel markers that may more accurately predict TMZ response. METHODS/STUDY POPULATION: We applied a combination of omics approaches and functional studies to a cohort of GBM patients to search for novel markers that would predict the response to TMZ treatment more accurately than traditional markers. Using a set of 47 primary and secondary GBM tumor samples, we employed comparative transcriptomics, whole exome sequencing, data independent acquisition (DIA) proteomics, and phosphoproteomics to look for DNA mutations and changes in gene expression and/or protein expression that correlated to response or non-response to TMZ. Subsequently, we performed functional studies and analyzed patient treatment data to validate our results. RESULTS/ANTICIPATED RESULTS: This study is in early stage, but we anticipate that our combination of methods may allow us to identify and validate at least one novel biomarker for TMZ response in patient GBM. For example, comparative transcriptomics or phosphoproteomics may identify a previously unrecognized gene or protein over/under-expression in a subset of patients. In this case we will validate findings using western blotting, IHC staining, and through siRNA of target gene/protein on patient derived GBM cells to examine if removal of this marker leads to TMZ sensitivity. We will further confirm prediction of marker correlation through comparison with matched patient treatment data. DISCUSSION/SIGNIFICANCE: The identification of improved biomarkers to predict response to TMZ treatment is a discovery that could rapidly become standard of care for GBM patients. It would ensure that all responders receive TMZ and avoid exposing nonresponders unnecessarily to TMZ and its potential side effects.
325 Predictors of Substance Use Initiation by Late Childhood: Findings from the Adolescent Brain Cognitive Development (ABCD) Study
Authors: Green ReJoyce , Kirkland Anna E. , Browning Brittney D. , Bryant Brittany E. , Garcia Alexis M. , Tomko Rachel L. , Gray Kevin M. , Mewton Louise , Wolf Bethany J. , Ferguson Pamela L. , Squeglia Lindsay M.
Published: April 2023

OBJECTIVES/GOALS: Adolescence represents a critical period for substance use initiation. Various factors may contribute to trying a sip or single puff of a substance, that could lead to more frequent use. However, less is known about how predictors from multiple domains converge to impact risk for general substance use initiation. METHODS/STUDY POPULATION: The Adolescent Brain Cognitive Development (ABCD) study is a multi-site longitudinal study following youth into early adulthood. The present study included 7,644 ABCD children who reported no lifetime substance use (including any experimentation) at baseline (ages 9–10). Our primary aim was to use a random forest classification model to predict binary substance use initiation, defined as trying any non-prescribed substance (e.g., alcohol, tobacco, cannabis, non-prescribed medications), during a 2-year follow-up after baseline. A total of 402 variables from the following categories were examined as predictors: demographics, peer substance use and availability, mental and physical health, culture and environment, biospecimens, neurocognitive functioning, and structural neuroimaging variables. RESULTS/ANTICIPATED RESULTS: Over a two-year follow-up, 751 (9.8%) of substance-naÃ¯ve children reported trying a substance by age 11. The most common substance was alcohol, followed by cannabis and tobacco. Mean Decrease Accuracy (MDA) values were used to assess the relative importance of each predictor. The overall accuracy of the model in accurately predicting group membership (no substance use initiation vs. substance use initiation) was 57.66%. Of the top 5 predictors, the most important predictor was intent to use alcohol (MDA = .002). The following top predictors were structural neuroimaging variables: volume and surface area of right lateral occipital lobe (MDA = .0009 and .0008, respectively), surface area of right inferior temporal lobe (MDA = .0007), and surface area of left superior frontal lobe (MDA = .0007). DISCUSSION/SIGNIFICANCE: A combination of intent to use alcohol and structural neuroimaging indices were among the top predictors of substance use initiation. Understanding predictors of early substance use experimentation is important for identifying at-risk youth that may require targeted intervention approaches.
349 Translation of novel multidisciplinary health technologies in the Ontario healthcare system: A case study of pharmacogenomic testing
Authors: Neumark Samuel , Schmitz Mary , Foty Richard , Ferenbok Joseph
Published: April 2023

OBJECTIVES/GOALS: There is a need for high-quality and efficient translation of health technologies in the Ontario healthcare system. The goal of this project is to understand the decision-making processes of government expert groups developing recommendations for the system-level implementation of pharmacogenomic testing. METHODS/STUDY POPULATION: This prospective observational case study includes the Ontario Health Pharmacogenomics (PGx) Working Group focused on developing recommendations for a PGx testing implementation strategy in the province. Ontario Health is the government agency that oversees provincial healthcare planning and service delivery. Using qualitative ethnographic methods, we will observe and document the working group’s activities over a 10-month period. Data collection involves meeting recordings, correspondences, researcher field notes, decision-making processes, and group characteristics. Using descriptive statistics and inductive qualitative analyses, the data will be examined to build theory and frameworks for knowledge translation. RESULTS/ANTICIPATED RESULTS: The results will be presented through a case report, process maps, decision milestones, visualizations, and procedural recommendations for future expert groups. This study will contribute to the body of foundational knowledge about translational sciences and support the National Center for Advancing Translational Sciences’guiding principles. To enhance translational processes and train the future translational workforce, this research can be used for educational initiatives. In addition, the observed processes will inform a theory about how expert recommendations are developed in public healthcare systems. DISCUSSION/SIGNIFICANCE: This research addresses a current gap in understanding around translational processes, government decision-making, and the development of recommendations for the adoption, implementation, and dissemination of the novel health technologies transforming public healthcare in Canada.
364 Beneficial Actions of SGLT2 Inhibition and H2S Therapy in Heart Failure with Preserved Ejection Fraction
Authors: Doiron Jake , Li Zhen , Xia Huijing , Lapenna Kyle B. , Allerton Timothy D. , Kapusta Daniel R. , Lefer David J.
Published: April 2023

OBJECTIVES/GOALS: SGLT2i therapy is currently a cornerstone in heart failure with preserved ejection fraction (HFpEF) therapy. Similarly, H2S has been shown to be beneficial in preclinical models of heart failure. With this in mind, we sought to investigate the effects of the SGLT2i and H2S donor therapy alone or in combination in a rodent model of cardiometabolic HFpEF. METHODS/STUDY POPULATION: Male C57BL/6N mice (9 weeks of age) were fed a high fat, Western diet (HFD) and received L-NG-Nitro arginine methyl ester (L-NAME) in the drinking water (0.5 g/L) to induce HFpEF. At 5 weeks, animals were randomized to either control, H2S donor (SG-1002, 90 mg/kg/d, P.O), Empagliflozin (155 mg/L, P.O), or the combination of SG-1002 and Empagliflozin for an additional 5 weeks while being maintained on HFD and L-NAME. Echocardiography, left ventricular invasive LV and systemic hemodynamics, and exercise capacity testing were performed to assess cardiovascular disease severity. Fasted glucose, circulating triglyceride and cholesterol content were similarly measured to quantify key clinical metabolic parameters. H2S and its metabolite, sulfane sulfur, were quantified to assure adequate H2S donation. RESULTS/ANTICIPATED RESULTS: Administration of SG-1002 restored H2S and sulfane sulfur to normal circulating levels. All treatment groups exhibited similar improvements in LV diastolic dysfunction as measured by E/E’and LVEDP. Combination therapy significantly improved exercise capacity whereas the monotherapy groups did not. Treatment with SG-1002 decreased fasting glucose and circulating cholesterol while all treatment groups displayed decreased circulating triglycerides and body weight compared to HFpEF control. DISCUSSION/SIGNIFICANCE: These data indicate that restoring H2S or treatment with an SGLT2i in this preclinical HFpEF model attenuates pathology. Combination of both drugs exhibited greater benefit than either monotherapy in important HFpEF parameters such as exercise capacity. Further studies are underway to characterize the benefits observed from combination therapy.
370 Epicardial adipose tissue and cardiometabolic health in youth-onset type 2 diabetes undergoing vertical sleeve gastrectomy
Authors: Dobbs Tyler , Kelsey Megan , Cree-Green Melanie , Baumgartner Amy , Bailey Alex , Gross Susan , Pyle Laura , Inge Thomas , Bjornstad Petter , Nadeau Kristen
Published: April 2023

OBJECTIVES/GOALS: The goal of this study is to investigate the potential independent relationship between epicardial adipose tissue (EAT) and cardiometabolic health in youth-onset type 2 diabetes (T2D) and explore changes in EAT as a potential mediator of changes in cardiometabolic health in response to vertical sleeve gastrectomy (VSG). METHODS/STUDY POPULATION: We will assess glycemic control, insulin sensitivity and secretion in youth with T2D before and 3 months after VSG. Fasting labs, anthropometrics, and a 4-hour, frequently sampled liquid mixed meal tolerance test (45g carbohydrates, 14g fat, and 14g protein) were performed. Calculations included glucose, insulin, and GLP-1 area under the curve (AUC), Matsuda Index, HOMA-IR, and oral disposition index (DI). These cardiometabolic outcomes will then be assessed for associations between total EAT volume, measured from cardiac MRI. RESULTS/ANTICIPATED RESULTS: Previous studies have shown that individuals with obesity have higher EAT than lean controls, and adults with T2D have even higher EAT than obese controls. Therefore, we anticipate that our participants will have higher volume of EAT than what has been reported in the literature and that they will have worsening cardiometabolic outcomes without MBS. Our anticipated results will include: Weight and BMI, hemoglobin A1c, diabetes medications, Matsuda Index, HOMA-IR, DI, and glucose and insulin AUC during an MMTT. Cardiac MRI's are being analyzed and will give total EAT volume and will be analyzed for correlations with the cardiometabolic outcomes of body composition, aortic stiffness, blood pressure, cardiac structure and function, as well as lipid panel and insulin sensitivity. DISCUSSION/SIGNIFICANCE: This study is the first to specifically assess EAT in adolescents with T2D. The assessment of EAT will be done with gold-standard MRI and correlated with cardiometabolic health assessed by gold-standard methods. Together, the results will give insight into EAT as a potential independent cardiometabolic risk factor in adolescents undergoing VSG.
390 A Mixed Methods Review of Endpoint Measures Utilized in Adult ADHD Medication Clinical Trial Design
Authors: Davies Claire , Church Terry David
Published: April 2023

OBJECTIVES/GOALS: To evaluate the clinical trial designs of stimulant medications approved for the treatment of adult Attention-Deficit/Hyperactivity Disorder (ADHD) and identify discrepancies between safety and efficacy outcome measure criteria. METHODS/STUDY POPULATION: There are 24 stimulants currently approved and marketed for the treatment of adult ADHD: 12 amphetamine (AMP)-based and 12 methylphenidate (MPH)-based formulations. A mixed methods review of clinical trials that have evaluated the safety and efficacy of these drugs will be performed using qualitative and quantitative data, including inclusion and exclusion criteria, primary and secondary outcome measures, manufacturer recommendations, and package inserts, as well as FDA recommendations for industry. After compiling a list of clinical trials for each of the 24 drugs, this information will be analyzed for themes and trends in the assessment of safety and efficacy with particular attention paid to differing criteria between individual drugs and/or the two subclasses of prescription stimulants. RESULTS/ANTICIPATED RESULTS: The FDA has published recommendations for ADHD medication clinical trial design, including suggested outcome measures utilizing validated assessment tools. However, the FDA notes that these assessments may be altered to fit study populations and indicate that other primary and secondary endpoint measures may be acceptable. As such, efficacy assessments for these drugs may vary greatly, and the specific criterion selected for each drug’s study population and outcome measures may suggest more specific indications and usage conditions for optimal efficacy. DISCUSSION/SIGNIFICANCE: Regulatory officials consider AMP- and MPH-based stimulants equivalent therapeutic options for the treatment of ADHD. A study of clinical trial criteria reveals that differing mechanisms have been used to evaluate safety and efficacy. This discrepancy may have implications that affect clinical decision-making and patient experiences.
429 Eribulin Synergizes with STING Agonists by Enhancing Type 1 Interferon Expression and Improves Antitumor Efficacy as Combination Treatment
Authors: Takahashi-Ruiz Leila , Fermaintt Charles S. , Wilkinson Nancy , Chan Peter Y. , Mooberry Susan L. , Risinger April L.
Published: April 2023

OBJECTIVES/GOALS: Triple-negative breast cancer (TNBC) is a subtype of breast cancer that lacks effective targeted treatment options. TNBC's greater degree of immunogenicity than other breast tumors makes immunotherapy a viable strategy. Strategies to improve the immunotherapy response includes targeting the cGAS-STING innate immune pathway with STING agonists. METHODS/STUDY POPULATION: We have previously shown in vitro that eribulin, a microtubule destabilizer currently used in the treatment of TNBC, functions as an indirect STING agonist because it promotes the release of mitochondrial DNA into the cytoplasm. Separately, eribulin also significantly enhances type I interferon expression induced by STING agonists measured by qRT-PCR through a second TBK1-dependent mechanism downstream of STING activation through detecting higher amounts of phosphorylated IRF-3 by western blot protein analysis. Mechanisms of eribulin-mediated interferon expression occur in immune and TNBC cells and are shared with other microtubule destabilizers but not with the microtubule stabilizing agent paclitaxel. RESULTS/ANTICIPATED RESULTS: We determine that the enhancement of type I interferon expression by eribulin is pharmacologically synergistic with multiple STING agonists. The significant enhancement by eribulin led us to evaluate the antitumor efficacy of eribulin in combination the STING agonist ADU-S100 in a challenging spontaneous mammary tumor model MMTV-PyVT. We show that the combination treatment significantly decreased tumor growth which allowed for longer survival compared to other groups. This is particularly interesting because of our previous studies showing that eribulin alone, but not paclitaxel, promotes the activation of CD4+ T-cells in the spleen and draining lymph nodes of BALB/c mice with 4T1 tumors through flow cytometric analysis. DISCUSSION/SIGNIFICANCE: These data contribute to accumulating evidence that there are important mechanistic differences between the microtubule targeted chemotherapeutics currently used in the treatment of TNBC and suggest that eribulin can act as an immune adjuvant in addition to its anti-mitotic effect.
443 Team Science
Authors: Miah Sayem , Acharya Baku , Huda Md Nazmul , Frett Brendan
Published: April 2023

OBJECTIVES/GOALS: T develop a novel PROTAC to deplete BRK to inhibit tumorigenesis and metastasis, which is unattainable by using tradition kinase inhibitors. METHODS/STUDY POPULATION: We will design, synthesize, and evaluate BRK-specific PROTACs to study both catalytic and scaffolding properties of BRK that contribute to cancer progression. Optimal PRTOAC concentrations will be used to treat MDA-MB-231 cells to determine effects on cell proliferation, cell migration, invasion, metastatic potential, and colony formation. Immunoblotting will be used to determine target protein degradation and to evaluate if PROTAC mediated degradation of BRK allows SMAD4 to form complex with SMAD2 and SMAD3. Finally, a metastatic xenograft mouse model (MDA-MD-231) will be injected subcutaneously or via tail vein into NU/J 002019 mice8, 17 and treated with the BRK PROTAC to evaluate inhibition of tumorigenesis and metastasis. RESULTS/ANTICIPATED RESULTS: We expect that the PROTAC will specifically degrade BRK and restore the anti-tumorigenic and anti-metastatic function of SMAD4 in metastatic breast cancer. In turn, we anticipate that cell proliferation, invasion, and colony formation properties of metastatic breast cancer will be restricted and SMAD4 will form complex with SMAD2 and SMAD3. Additionally, mice treated the BRK targeted PROTAC should experience reduced tumor growth and metastasis compared to placebo. DISCUSSION/SIGNIFICANCE: This substantially new approach will inhibit oncogenic nRTKs to restore the antitumor function of TGFβ/SMAD signaling is expected to bring metastatic cancer under therapeutic control. Thus, the expected outcomes are likely to have a significant impact because it will provide a new targeted therapy to improve metastatic TNBC patient survival.
445 The effects of dietary fiber based on fermentability and viscosity on phosphorus absorption and the gut microbiome in chronic kidney disease-mineral and bone disorder
Authors: Biruete Annabel , Chen Neal X. , Srinivasan Shruthi , O'Neill Kalisha , Nelson David , Hill Gallant Kathleen M. , Moe Sharon M.
Published: April 2023

OBJECTIVES/GOALS: To compare the effects of dietary fiber supplementation based on fermentability and viscosity on phosphorus fractional absorption and the gut microbiome in a rat model of chronic kidney disease-mineral and bone disorder (CKD-MBD). METHODS/STUDY POPULATION: 25-week-old Cy/+ male rats (CKD hereafter) will be randomly assigned to receive one of four fiber treatments (10% w/w each) based on fermentability and viscosity: 1) Cellulose (-fermentability, -viscosity), 2) inulin (+fermentability, -viscosity), 3) psyllium husk (-fermentability, +viscosity), or 4) pectin (+ fermentability, +viscosity). Diets will be formulated with a semipurified diet containing 0.7% phosphorus. Treatments will last for 10 weeks, and rats will be euthanized at 35 weeks of age, where animals have reached kidney failure. Intravenous and oral 33P will be used for intestinal phosphorus fractional absorption and cecal/fecal samples will be obtained at euthanasia for microbiome assessment using shotgun metagenomics. RESULTS/ANTICIPATED RESULTS: Our preliminary data show that fermentable dietary fiber (inulin) impacted phosphorus homeostasis by increasing the circulating levels of fibroblast growth factor-23 (a bone-derived hormone that increases phosphorus excretion in urine) and lowering circulating levels of phosphorus in the Cy/+ male rat model of progressive chronic kidney disease. We hypothesize that dietary fiber impacts phosphorus absorption in gut microbiome-dependent and independent mechanisms. For example, fermentable fiber enhances the production of short-chain fatty acids, lowering the intraluminal pH, and enhancing mineral solubility and absorption. Meanwhile, viscous fibers may encapsulate minerals limiting their absorption if these fibers are non-fermentable. DISCUSSION/SIGNIFICANCE: Hyperphosphatemia, or high circulating phosphorus, is a major factor in the pathogenesis of CKD-MBD. Treatment of hyperphosphatemia is focused on reducing intestinal absorption. However, available therapies vary in their efficacy and focus on phosphorus absorption in the small intestine, ignoring the possible impact of the large intestine.
464 Chronic HIV infection influences the immune response during acute COVID-19 and long COVID
Authors: Opsteen Skye , Fram Tim , Long Dustin , Erdmann Nathan
Published: April 2023

OBJECTIVES/GOALS: Despite highly effective antiretroviral therapy, people living with HIV (PLWH) experience chronic immune activation and inflammation which may influence the progression of infections such as SARS-CoV-2. Here, we explore the immune response and clinical outcomes in HIV(+) and HIV(-) individuals experiencing acute COVID-19 and long COVID (LC). METHODS/STUDY POPULATION: We performed flow cytometric analyses on peripheral blood mononuclear cells from the following: 1) HIV(-) individuals experiencing acute COVID-19, 2) PLWH experiencing acute COVID-19, and 3) pre-COVID-19 pandemic PLWH. Additionally, we will perform similar analyses for the following: 1) PLWH experiencing LC, 2) PLWH previously infected with SARS-CoV-2 who recovered, 3) pre-COVID-19 pandemic PLWH, and 4) HIV(-) individuals experiencing LC. Flow cytometry panels include surface markers for immune cell populations, activation and exhaustion surface markers (with and without SARS-CoV-2-specific antigen stimulation), and intracellular cytokine staining. We will also analyze how chronic HIV infection and other clinical and demographic factors (e.g., age, CD4 %) impact persistent symptomatic burden. RESULTS/ANTICIPATED RESULTS: Acute COVID-19 results–Overall, PLWH had higher baseline expression of activation markers OX40 and CD137 on CD4+ and CD8+ T cells, along with increased levels of TNFa producing CD8+ T cells. Interestingly, PLWH had increased expression of exhaustion markers PD1 and TIGIT but decreased expression of TIM3 on CD4+ and CD8+ T cells. Additionally, PLWH had decreased levels of IL-2 and IFNg producing CD4+ T cells which suggests functional exhaustion. Long COVID-19 expected results–we hypothesize that the activation and inflammation seen in chronic HIV infection will lead to more immune dysregulation and subsequently worsened symptomatic burden. Additionally, we hypothesize that PLWH may have different frequencies of certain LC manifestations, such as increased rates of neurocognitive impairment. DISCUSSION/SIGNIFICANCE: Our findings suggest that chronic HIV infection influences acute immune response during SARS-CoV-2 infection, and that PLWH have variable expression of exhaustion markers which warrants further study. Additionally, our findings in the LC cohort will aid in characterizing clinical manifestations and immunologic mechanisms of LC in PLWH.
485 Modeling gastric mucus layer physiology using human organoids
Authors: Lyon Katrina , Sidar Barkan , Dudiak Cam , Wilking Jim , Bansil Rama , Bimczok Diane
Published: April 2023

OBJECTIVES/GOALS: Our goal is to explore the extent to which organoids can serve as models for the protective mechanisms of the stomach–the mucus barrier and the pH gradient across it. We aim to first optimize and validate an organoid-based model of the gastric mucus layer, and then define the cellular mechanisms by which the gastric pH gradient is maintained across it. METHODS/STUDY POPULATION: We have developed a method for the in vitro engineering of gastric mucus by growing epithelial cells at the air-liquid interface (ALI). We use microrheology with fluorescent microspheres to define and compare the biophysical and viscoelastic properties of our lab-grown mucus to those of native mucus. We will perform CryoFE-SEM to compare the internal heterogeneity of our lab-grown mucus to fresh mucus obtained from patient tissue. For our mechanistic studies, we will use a pH-sensitive dye (methyl red) to assess the ability of our lab-grown mucus to maintain an artificial pH gradient in a microfluidic device. Next, we will use a pH microelectrode to measure proton flux through our mucus in vitro, investigating the potential for a physiological gradient in both 2D and 3D organoid models. RESULTS/ANTICIPATED RESULTS: Here we show that gastric organoids and their corresponding epithelial monolayers produce a mucus gel that does indeed mimic in vivo functions. Immunohistochemical staining, electron microscopy, microrheology, and particle tracking analyses revealed that our gastric organoid mucus is viscoelastic and structurally heterogeneous–both properties that are crucial to the stomach’s mucosal first line of defense. Mechanically similar mucus was also engineered using two-dimensional air-liquid interface cultures of the same epithelia. Lastly, live confocal imaging revealed that H. pylori motility–an important virulence factor–was drastically hindered by our lab- grown mucus. DISCUSSION/SIGNIFICANCE: We describe a novel method for the in vitro engineering of gastric mucus and highlight biophysical properties that contribute to our stomach’s defense against pathogens. This work will lead to an improved understanding of gastric physiology and may contribute to the development of novel drug delivery systems to tackle diseases of the gastric mucosa.
491 Outcomes of an Integrated Research Ethics Consultation Service
Authors: Smith Elise , Farroni Jeffrey S. , McNamara Victoria H.
Published: April 2023

OBJECTIVES/GOALS: The need for mechanisms of ethical discourse and guidance has increased as translational research collaborations become more complex. The goal of this project is to analyze the stakeholder engagement and ethical issues our research ethics consultation service (RECS) conducted over a two year period. METHODS/STUDY POPULATION: We conducted a retrospective review of our RECS database from 2020 to 2022. We examined the nature of the research and ethical issues of concern from consult requestors, including whether or not consults were preventative. In addition, we assessed the educational outreach conducted during that timeframe as a measure of service awareness. RESULTS/ANTICIPATED RESULTS: There was a total of 42 consults conducted over the previous year. There were a wide variety of issues related to informed IRB-related processes (31%), consent (24%), QA/QI determination (12%), authorship (10%), confidentiality (7%), diversity/inclusion (7%), grant preparation (7%). Many of the consults (n=28, 67%) included secondary issues. A few consults (n=4, 10%) were preventative, meaning that the consult was requested in anticipation or consideration of a potential ethical issue. Outreach efforts extended to a diverse array of institutional stakeholders and trainees. DISCUSSION/SIGNIFICANCE: The RECS serves numerous constituencies throughout our institution on ethical issues spanning nearly all aspects of research design, conduct, and analysis. These data highlight initiatives to increase study efficiency (in collaboration with institutional research oversight) and helps to direct educational efforts and outreach.
The Extent and Duration of Primary Schooling in Eighteenth-Century America
Authors: Shammas Carole
Published: April 2023

The educational attainment literature has brought back interest in early American primary schools, and much current research views those schools as superior to their European peers in the education offered to youth. Its emphasis, though, on using school enrollment as the prime indicator of attainment conflicts with the revisionist view of a previous generation of historians who argued that education in the heavily rural and agricultural society of the time should be considered as a process of social reproduction delivered by households, with schools being peripheral for most youth. This article, relying on evidence from statutes, indentures, and a 1798 New York State school survey, finds increased resort to primary schooling over the eighteenth century, attributable not to American exceptionalism but to a transatlantic movement away from scribal-dominated literacy and numeracy toward common use of a standardized written vernacular and “arithmetic by pen.” However, the dependence of households on child labor meant that the Three Rs did not get distributed in either an egalitarian or compact fashion. Small doses spread over a number of years—educational sprawl—best describes the system, and it lasted through much of the nineteenth century.
Turbulent thermal convection in mixed porous–pure fluid domains
Authors: Hamtiaux Victoria , Papalexandris Miltiadis V.
Published: April 2023

In this paper, we report on a direct numerical simulation (DNS) study of turbulent thermal convection in mixed porous–pure fluid domains. The computational domain consists of a cavity that contains a porous medium placed right above the bottom wall. The solid matrix is internally heated which, in turn, induces the convective motions of the fluid. The Rayleigh number of the flow in the pure fluid region above the porous medium is of the order of

$10^7$

. In our study, we consider cases of different sizes of the porous medium, as well as cases with both uniform and non-uniform heat loading of the solid matrix. For each case, we analyse the convective structures in both the porous and the pure fluid domains and investigate the effect of the porous medium on the emerging flow patterns above it. Results for the flow statistics, as well as for the Nusselt number and each of its components, are also presented herein. Further, we make comparisons of the flow properties in this pure fluid region with those in Rayleigh–Bénard convection. Our simulations predict that, depending on the area coverage, the large-scale circulation above the porous medium can be in a single-roll, dual-roll or intermediate state. Also, when the area coverage increases, the temperatures inside it increase due to reduced fluid circulation. Accordingly, when the area coverage increases, then the Nusselt number becomes smaller whereas the Rayleigh number is increased.
Turbulent disruption of density staircases in stratified shear flows
Authors: Petropoulos Nicolaos , Mashayek Ali , Caulfield Colm-cille P.
Published: April 2023

The formation of step-like ‘density staircase’ distributions induced by stratification and turbulence has been widely studied, and can be explained by the ‘instability’ of a sufficiently strongly stably stratified turbulent flow due to the decrease of the turbulent density flux with increasing stratification via the ‘Phillips mechanism’ (Phillips, Deep Sea Research and Oceanographic Abstracts, vol. 19, 1972, pp. 79–81. Elsevier). However, such density staircases are not often observed in ocean interiors, except in regions where double-diffusion processes are important, leading to thermohaline staircases. Using reduced-order models for the evolution of velocity and density gradients, we analyse staircase formation in stratified and sheared turbulent flows. Under the assumption of inertial scaling

$\epsilon \sim U^3/L$

for the kinetic energy dissipation rate

$\epsilon$

, where

$U$

and

$L$

are characteristic velocity and length scales, we determine ranges of bulk Richardson numbers

${Ri}_{b}$

and turbulent Prandtl numbers

${Pr}_{T}$

for which staircases can potentially form and show that the Phillips mechanism only survives in the limit of sufficiently small turbulent Prandtl numbers. For relevant oceanic parameters, a range of turbulent Prandtl numbers above which the system is not prone to staircases is found to be

${Pr}_T \simeq 0.5\unicode{x2013}0.8$

. Since several studies indicate that the turbulent Prandtl number in stably stratified turbulence and in ocean interiors is usually above this threshold, this result supports the empirical observation that staircases are not favoured in ocean interiors in the presence of relatively homogeneous and sustained turbulence. We also show that our analysis is robust to other scalings for

$\epsilon$

(such as the more strongly stratified scaling

$\epsilon \sim U^{2}N_{c}$

, where

$N_{c}$

is a characteristic value of the buoyancy frequency), supporting our results in both shear-dominated and buoyancy-dominated turbulent regimes as well as in weakly and strongly stratified regimes.
Recovery of steady rotational wave profiles from pressure measurements at the bed
Authors: Clamond Didier , Labarbe Joris , Henry David
Published: April 2023

We derive equations relating the pressure at a flat seabed and the free-surface profile for steady gravity waves with constant vorticity. The resulting set of nonlinear equations enables the recovery of the free surface from pressure measurements at the bed. Furthermore, the flow vorticity (unknown a priori) is determined solely from the bottom pressure as part of the recovery method. This approach is applicable even in the presence of stagnation points and its efficiency is illustrated via numerical examples.
Usual intake of one-carbon metabolism nutrients in a young adult population aged 19–30 years: a cross-sectional study
Authors: Jindasereekul Phachara , Jirarattanarangsri Wachira , Khemacheewakul Julaluk , Leksawasdi Noppol , Thiennimitr Parameth , Taesuwan Siraphat
Published: April 2023

One-carbon nutrients play an important role in epigenetic mechanisms and cellular methylation reactions. Inadequate intake of these nutrients is linked to metabolic perturbations, yet the current intake levels of these nutrients have rarely been studied in Asia. This cross-sectional study surveyed the usual dietary intake of one-carbon nutrients (folate, choline and vitamins B2, B6 and B12) among Thai university students aged 19–30 years (n 246). Socioeconomic background, health information, anthropometric data and 24-h dietary recall data were collected. The long-term usual intake was estimated using the multiple-source method. The average usual intake levels for men and women were (mean ± sd) 1⋅85 ± 0⋅95 and 2⋅42 ± 8⋅7 mg/d of vitamin B2, 1⋅96 ± 1⋅0 and 2⋅49 ± 8⋅7 mg/d of vitamin B6, 6⋅20 ± 9⋅5 and 6⋅28 ± 12 μg/d of vitamin B12, 195 ± 154 and 155 ± 101 μg dietary folate equivalent/d of folate, 418 ± 191 and 337 ± 164 mg/d of choline, respectively. Effect modification by sex was observed for vitamin B2 (P-interaction = 0⋅002) and choline (P-interaction = 0⋅02), where every 1 mg increase in vitamin B2 and 100 mg increase in choline intake were associated with a 2⋅07 (P = 0⋅01) and 0⋅81 kg/m2 (P = 0⋅04) lower BMI, respectively, in men. The study results suggest that Thai young adults meet the recommended levels for vitamins B2, B6 and B12. The majority of participants had inadequate folate intake and did not achieve recommended intake levels for choline. The study was approved by the Ethics Committee at the Faculty of Medicine, Chiang Mai University. This trial was registered at www.thaiclinicaltrials.gov (TCTR20210420007).
Characterization theorems for pseudo cross-variograms
Authors: Dörr Christopher , Schlather Martin
Published: April 2023

Pseudo cross-variograms appear naturally in the context of multivariate Brown–Resnick processes, and are a useful tool for analysis and prediction of multivariate random fields. We give a necessary and sufficient criterion for a matrix-valued function to be a pseudo cross-variogram, and further provide a Schoenberg-type result connecting pseudo cross-variograms and multivariate correlation functions. By means of these characterizations, we provide extensions of the popular univariate space–time covariance model of Gneiting to the multivariate case.
20 YEARS OF AMS 14C DATING USING THE ARTEMIS FACILITY AT THE LMC14 NATIONAL LABORATORY: REVIEW OF SERVICE AND RESEARCH ACTIVITIES
Authors: Beck L , Caffy I , Delqué-Količ E , Dumoulin J-P , Goulas C , Hain S , Moreau C , Perron M , Setti V , Sieudat M , Thellier B
Published: April 2023

In 2001, five French public organizations (CNRS, CEA, IRD, IRSN, and the Ministère de la Culture) signed an agreement to purchase a new accelerator mass spectrometer to provide radiocarbon dating services at the national level. The Laboratoire de Mesure du Carbone 14 (LMC14) was set up in Saclay (France) around ARTEMIS, an AMS system based on a 3MV Pelletron from NEC and installed in early 2003. In 2015, the LMC14 joined the Laboratoire des Sciences du Climat et de l’Environnement, making it possible to develop research projects in addition to the service activity and since 2021, the LMC14 has been a member of the IAEA Collaborating Centre “Atoms for Heritage” at the Université Paris-Saclay. Since 2003, 70,000 samples have been measured. Two-thirds of the samples have been prepared on site and one-third in two associated laboratories in Paris and Lyon. Over the past years, the LMC14 has participated in several international inter-comparisons and has continuously improved its capabilities by developing new protocols for preparation and measurement. In this paper, the radiocarbon dating services of the last 20 years for research institutions, museums and environmental monitoring are reviewed and recent results from environmental and archaeological research programs are highlighted.
Lexical Ambiguity in Political Rhetoric: Why Morality Doesn't Fit in a Bag of Words
Authors: Kraft Patrick W. , Klemmensen Robert
Published: April 2023

How do politicians use moral appeals in their rhetoric? Previous research suggests that morality plays an important role in elite communication and that the endorsement of specific values varies systematically across the ideological spectrum. We argue that this view is incomplete since it only focuses on whether certain values are endorsed and not how they are contextualized by politicians. Using a novel sentence embedding approach, we show that although liberal and conservative politicians use the same moral terms, they attach diverging meanings to these values. Accordingly, the politics of morality is not about the promotion of specific moral values per se but, rather, a competition over their respective meaning. Our results highlight that simple dictionary-based methods to measure moral rhetoric may be insufficient since they fail to account for the semantic contexts in which words are used and, therefore, risk overlooking important features of political communication and party competition.
Bordering the GDR: Everyday Transnationalism, Global Entanglements and Regimes of Mobility at the Edges of East Germany
Authors: Richardson-Little Ned , Stokes Lauren
Published: April 2023

No state has ever been as identified with its borders as the German Democratic Republic (GDR). The guest editors’ introduction to this special issue analyzes the development of the historiography of the borders of the GDR, showing how new approaches to the country's history have also impacted scholarship on the everyday history of the border. We argue for approaches that understand the border simultaneously as a site of conflict and cooperation and that situate the border not just alongside its geographical neighbors, but within broader flows of natural resources, pollution, narcotics, migration, and disease. Drawing on the interdisciplinary field of border studies, we argue that global approaches can help contextualize the exceptional and encourage scholars to ask new questions about which elements of GDR bordering practices were part of the globally emerging normalcy of border regimes, and which were unique to East Germany. In these ways, this special issue seeks to reveal new aspects of East German history and, in turn, make the GDR more legible within border studies.
Racial Profiling on the U-Bahn: Policing the Berlin Gap in the Schönefeld Airport Refugee Crisis
Authors: Stokes Lauren
Published: April 2023

The German Democratic Republic (GDR) treated the Berlin Wall as an official state border, but the Federal Republic of Germany (FRG) did not recognize it as an official state border and thus did not impose entry controls. This asymmetric recognition opened up a gap in the regime of border policing and turned divided Berlin into one of the most significant sources of unauthorized migration into the FRG, creating tensions in Berlin, West Germany, and western Europe more broadly. Countries including France, Denmark, and the Netherlands all pressured the FRG to shut the open border in Berlin. This article examines how West German authorities sought to respond to their demands without recognizing the Berlin Wall as a state border. West German authorities pursued two broad strategies. The first involved internalizing the border through institutionalized racial profiling in West Berlin. The second entailed externalizing the border by asking the GDR to enforce FRG visa and passport requirements. Although both forms of border policing have often been associated with the end of the Cold War, this article shows that they were adopted earlier, and in response to Cold War imperatives.
Multiple datasets confirm range extension of the sicklefin devil ray Mobula tarapacana in the western North Atlantic Ocean off the eastern USA
Authors: Pate Jessica H. , Wilmott Julia R. , Jones Christian , Horn Calusa , Farmer Nicholas A.
Published: April 2023

The sicklefin devil ray (Mobula tarapacana) is a large, pelagic ray which is listed as Endangered by the International Union for Conservation of Nature (IUCN). Mobula tarapacana is thought to have a circumglobal, yet patchy distribution, and has not been verified extant off the eastern USA. Here, we report 180 sightings of M. tarapacana with a total of 361 individuals, collated across five datasets from aerial survey operations and incidental sightings in the waters off the US East Coast and Gulf of Mexico, between 1996 and 2022. This study extends the northern range of M. tarapacana in the Gulf of Mexico to 29°N, and in the Atlantic to 40°N. Seasonal trends were observed off the north-eastern coast of the USA, with M. tarapacana only present in the summer months. Measurements from high resolution digital aerial imagery found M. tarapacana off the New York coast to be adults and subadults with an average disc width of 268 cm (±25, range 232–316 cm). This study provides important spatial and temporal data for management, as well as informing areas for future research on M. tarapacana in the western Atlantic.
‘East Punjab must not lag behind’: Partition, museums, and identity in independent India
Authors: Venkateswaran Mrinalini
Published: April 2023

This article foregrounds the postcolonial museum as a new source, and site, from which to write South Asian histories of partition and its aftermath. It focuses on collecting practices in India within East Punjab, following the partition of the British-era Punjab province in 1947 between India and Pakistan. Tapping hitherto-unused archival sources, it reveals the considerable financial investment and drive to collect at this time, belying the idea of museums being ‘dead’ colonial assets, and demonstrates their centrality to how citizenship and belonging were articulated (or withheld) in independent India. Some discoveries have far-reaching implications for both historians and museum professionals. The article also shines a light upon a new range of actors—both named and nameless, professional and citizen—who have been marginal to historical enquiry thus far. Moving beyond the familiar colonial templates within which museums in the region have until now been studied, it asks critical questions of thepostcolonialmuseum in South Asia by interrogating the relationship between collections, and the Indian nation-state and its subsidiaries.