bluuebunny/crossref_metadata_2025 huggingface dataset:
222 Stress and Coping Efforts Among Caregivers of Children with Down Syndrome: Application of The Transactional Model of Stress and Coping
Authors: Brantley Caroline , Knol Linda L. , Douglas Joy W. , Hernandez-Reif Maria , Lawrence Jeannine C. , Wind Stefanie A.
Published: April 2023

OBJECTIVES/GOALS: Stress is amplified in parents of children with Down syndrome (DS). However, little is known about caregivers’use of coping strategies in relation to stress-related eating and their child with DS. The purpose of this study is to explore stressors and coping resources that increase or decrease stress-related eating among caregivers of children with DS. METHODS/STUDY POPULATION: This descriptive qualitative study sought to understand and describe the lived experiences of caregivers of children with DS. Fifteen caregivers of children, ages 2-6-years with DS, completed an online survey and individual interview. The Transactional Model of Stress and Coping was used to develop the interview guide and to assess related constructs using the Mindful Eating Questionnaire (MEQ), Perceived Stress Scale (PSS), Salzburg Stress Eating Scale (SSES), and a subscale of the Eating and Appraisal Due to Emotions and Stress Questionnaire (EADES). Interviews were audio-recorded, transcribed verbatim, coded using NVivo 12, and analyzed using deductive thematic analysis based on theoretical constructs and content analysis approaches. RESULTS/ANTICIPATED RESULTS: Thirteen of the 15 participants reported moderate to high perceived stress on the PSS. Stressors identified by caregivers included caring for a child with DS, feeding difficulties, physical and mental health, family and social interactions, and finances. Caregivers utilized various interpersonal resources to cope, including family, friends, and social media groups. In the interviews, stress-related eating was revealed as a coping mechanism for 9 caregivers. Also, 13 caregivers reported eating more or eating less when stressed on the SSES and 7 caregivers reported increased emotional eating as measured on the sub-scale of the MEQ. Only two caregivers described using mindfulness strategies throughout the interview. DISCUSSION/SIGNIFICANCE: Overall, caregivers of children with DS report increased perceived stress related to their role as a caregiver of a child with DS, which may be related to changes in caregivers’dietary intake. Dietary interventions developed for these families should include mindful eating and stress management strategies.
224 The effect of housing status (homelessness vs. housed) on naloxone administration among patients with opioid overdose assessed by emergency medical services: A prehospital cross-sectional review
Authors: Abramson Tiffany M. , Abramson Corey M. , Sanko Stephen , Eckstein Marc , Wenzel Suzanne , Burner Elizabeth
Published: April 2023

OBJECTIVES/GOALS: People experiencing homelessness vs. housed peers have higher rates of substance use disorders as well as increased emergency medical services (EMS) use. However, it is unknown if EMS administers naloxone at different rates to people experiencing homelessness. We address if patient housing status impacts naloxone administration by EMS providers. METHODS/STUDY POPULATION: We conducted a retrospective, cross-sectional analysis of electronic patient care reports (ePCRs) for all 911-incidents attended by the Los Angeles Fire Department (LAFD) during the study period, January to December 2018. Individuals who were medically assessed by the LAFD and who were administered naloxone by LAFD EMS were included. Exclusion criteria was incomplete ePCRs. The primary outcome was prevalence of EMS naloxone administration by housing status in the city of Los Angeles. We used descriptive statistics and a logistic regression model to examine differences in care. RESULTS/ANTICIPATED RESULTS: Naloxone was administered in 2,438 of the 345,190 incidents that occurred during the study period. 608 (25%) incidents involved people experiencing homelessness. Top indications for naloxone administration were similar in both groups: overdose, altered consciousness and cardiac arrest. Of those who received naloxone, people experiencing homelessness were more likely to be male (82% v 67%) and younger (41 v 46 years). People experiencing homelessness were more likely to receive naloxone (OR 2.6, 95% CI 2.4-2.9). People experiencing homelessness received naloxone at a rate of 44 times that of housed peers. A logistic regression model adjusting for gender, age, respiratory depression and transport status showed people experiencing homelessness remained more likely to receive naloxone (OR 2.3, 95% CI 2.0-2.5). DISCUSSION/SIGNIFICANCE: Emergency medical services are more likely to administer naloxone to people experiencing homelessness than housed peers. There is a need to identify bias and factors that impact prehospital care and patient outcomes of people experiencing homelessness. New care pathways for people confronting homelessness and opioid use disorders are needed.
225 The Feasibility and Acceptability of a Remote Glucose Monitoring Program for Pregnant Marshallese Women with Pre-Gestational and Gestational Diabetes
Authors: Andersen Jennifer A. , McElfish Pearl , Su Dejun , Selig James , Schootman Mario , Eswaran Hari
Published: April 2023

OBJECTIVES/GOALS: The objective of the study is to test the feasibility and acceptability of a remote glucose monitoring program for pregnant Marshallese women with pre-gestational (PGDM) or gestational diabetes (GDM) without excluding women with limited English proficiency. The study will explore changes in patient-physician communication and self-efficacy. METHODS/STUDY POPULATION: Twenty Marshallese pregnant women with PGDM or GDM will be identified for recruitment. The patient will meet with a bilingual staff member to explain the study and provide consent. At enrollment, patients will complete an enrollment survey and be provided with the iGlucose monitor and receive training its use. Their provider(s) will receive access to their patient’s online portal for monitoring. At 14 days postpartum patients will receive a post-intervention survey and invitation to participate in interview, and their provider(s) will receive a satisfaction survey for the remote monitoring program. RESULTS/ANTICIPATED RESULTS: We hypothesize that the use of a remote glucose monitoring intervention is feasible and acceptable for Marshallese women with PGDM or GDM. Further, we hypothesize that the remote glucose monitoring program will increase satisfaction with physician-patient communication and will increase self-efficacy with glucose management. We anticipate that providers will find the program feasible and acceptable for use with the Marshallese population. DISCUSSION/SIGNIFICANCE: RPM technology can transform the way PGDM and GDM are managed. RPM may have greater benefits when there is a lack of language concordance by providing more time for patient-provider communication, thereby improving patient satisfaction and decreasing the risk of negative outcomes for Marshallese women.
233 Uganda-based Survey of Challenges in Breast Cancer Detection in Low and Middle Income Countries
Authors: Sathi Krishna Tejaswini , Yeo Kim Hwang , Naicker Pav , Pichay Leanne , Fuleihan Antony , Waiswa Peter , Yazdi Youseph
Published: April 2023

OBJECTIVES/GOALS: Low and middle income countries (LMICs) face challenges in early detection of breast cancer resulting in high breast cancer mortality. This study serves to identify gaps and opportunities for innovators seeking to address problems in early detection of breast cancer in Uganda and other LMICs. METHODS/STUDY POPULATION: Two methods were used: 1) Three weeks of ethnographic research in Uganda through primary stakeholder interviews and clinical observations. Interviews were conducted with patients, clinicians, NGOs, and key opinion leaders from the Uganda Cancer Institute, Makerere University, and JHPIEGO. Clinical observations were performed to note the workflow and availability of resources across diverse health centers ranging from village health teams in rural settings to the national referral hospital in the urban center. 2) A targeted literature search focused on breast cancer detection in LMICs. Keywords included breast cancer’, screening’, and diagnosis’. Identified challenges were validated through stakeholder interviews and categorized. Potential solutions to each challenge were explored. RESULTS/ANTICIPATED RESULTS: Three broad categories of challenges and suggested innovation targets were identified. 1) Ineffective clinical processes: deskilling and improving training around the process of clinical breast examinations, imaging operation and interpretation, and pathology preparation and interpretation; 2) Accessibility: increasing screening throughput, improving rural community access to breast cancer care, and increasing opportunistic screening; 3) Sensitization: increasing patient and health worker awareness of clinical presentations of breast cancer, reducing cultural barriers, and improving trust in the medical community. DISCUSSION/SIGNIFICANCE: Innovators seeking to solve problems in early breast cancer detection in LMICs should focus on ineffective clinical processes, accessibility, and sensitization. In conjunction with prompt treatment, there is potential to reduce breast cancer mortality rates in line with the Global Breast Initiative.
245 Do they REALLY Trust Us?: The Importance of Trust and Trustworthiness in All IN for Health
Authors: Sotto-Santiago Sylk , Hudson Brenda , Claxton Gina , Delp Lynsey , Lynch Dustin , Wiehe Sarah , Moe Sharon
Published: April 2023

OBJECTIVES/GOALS: The critical role that trust plays continues to be documented and highlights the gap in understanding the extent to which initiatives such as All IN for Health from the Indiana Clinical and Translational Institute (I-CTSI) can improve trust between the community and researchers. We discuss measures of trust and evaluate how we may improve recruitment. METHODS/STUDY POPULATION: In this study, we invited over 13000 volunteers from the All IN for Health research participant registry. Six hundred and sixty-three (663) respondents participated in the survey. The Relationship of Trust and Research Engagement Survey included three validated surveys: Distrust in Healthcare Organizations, the Trust in Medical Researchers scale by Hall et. al, and Patient Trust in Medical Researchers by Mainous et al.. The 36-item survey also included open-text questions. RESULTS/ANTICIPATED RESULTS: Based on preliminary results 74% agreed it’s safe to be in medical research, yet 79% had never been asked to participate in medical research by their doctor. Sixty percent believed that HCOs put money above patients’ needs. Forty percent agreed that doctors do medical research for selfish reasons. Fifty percent disagree that patients get the same medical treatment regardless of race/ethnicity. Moreover, 28 % agree that medical researchers act differently toward minorities. Between 9 and 11% believe that researchers select minorities for their most dangerous studies and some projects are secretly designed to expose minoritized groups to diseases. Our next step is to disaggregate the data by race and ethnicity and evaluate these answers. DISCUSSION/SIGNIFICANCE: This study's population willingly engaged in a research registry making their diminished trust quite alarming. Amongst the general population, trust in scientists is now below pre-pandemic levels. We must critically assess our own trustworthiness, and critically reflect on the authenticity of our efforts.
267 Defining the single-cell transcriptomes of splenic adaptive Natural Killer cells in donors with latent human cytomegalovirus infection
Authors: Khalil Mohamed , Terhune Scott , Malarkannan Subramaniam
Published: April 2023

OBJECTIVES/GOALS: The primary objective of this study was to define the transcriptomes and transcriptional regulatory network required for the development and function of adaptive Natural Killer (NK) cells in donors with latent human cytomegalovirus (HCMV) infection. METHODS/STUDY POPULATION: Eight healthy adult human spleens were obtained from four HCMV seropositive and four HCMV seronegative donors. Spleens were provided by the Versiti Organ Donor Center of Wisconsin and were processed to a single cell suspension. CD7+ CD3E- CD14- CD19- CD20- NK cells were isolated, using the BD FACSAria sorter. Following cell sorting, single-cell RNA sequencing (scRNA-seq) was performed, and cDNA libraries were constructed and sequenced via NextSeq 550. Cell Ranger was then used to algin the cDNA reads and the Seurat R package was used to analyze the transcriptional data. Cells were filtered and clustered based on the number of uniquely expressed genes. The monocle software was used for single cell trajectory analysis and the SCENIC software was used to decipher gene regulatory networks. RESULTS/ANTICIPATED RESULTS: Eight healthy spleens from four HCMV seropositive and four HCMV seronegative donors were obtained and their NK cells were sorted and captured for scRNA-seq. Donor median age was 59 [IQR 48.5-56.5], 50% (n=4) were female and all donors were not experiencing any acute or chronic symptoms. Using scRNA-seq, we observed elevated numbers of NKG2C+ adaptive NK cells in HCMV seropositive individuals when compared to HCMV seronegative individuals. In addition, we identify a set of transcription markers and regulators that are responsible for the development and function of adaptive NKG2C+ NK cells. Finally, our trajectory analysis of adaptive NKG2C+ NK cells revealed a unique developmental pathway. DISCUSSION/SIGNIFICANCE: Here, we demonstrate that HCMV infection can induce the formation of adaptive NKG2C+ NK cells that display a unique transcriptional and developmental profile. These findings have the potential to influence the future application of adaptive NK cells in cellular immunotherapies.
268 DEGAS: Deep transfer learning reveals cancer-like transcriptional signatures in histologically normal prostate tissue and adjacent-normal tissues in pancreatic cancer
Authors: Couetil Justin , Liu Ziyu , Zhang Jie , Huang Kun , Johnson Travis
Published: April 2023

OBJECTIVES/GOALS: Single-cell and spatial transcriptomics have revealed high heterogeneity in the tumor and microenvironment. Identifying populations of cells that impact a patient’s prognosis is an important research goal, so researchers can generate hypotheses and clinicians can provide targeted treatment. METHODS/STUDY POPULATION: DEGAS uses deep-transfer-learning to identify patterns between patient tumor RNA-seq and clinical outcomes and map these associations on to higher-resolution data like spatial and single-cell transcriptomics. We apply DEGAS to prostate and pancreatic cancer spatial transcriptomics samples, as well as one normal sample of prostate tissue. We used the TCGA prostate cancer cohort to with the accompanying survival information and publicly accessible prostate cancer ST data from 10X Genomics to predict survival associations in the ST slides derived from the TCGA patients. Based on these survival associations, we identify higher risk subsections of ST slides which can be further studied. RESULTS/ANTICIPATED RESULTS: We were able to validate our method by comparing it to Scissor and were able to show that the number of high-risk regions in prostate cancer slides increased with the stage of disease. Furthermore, we identify transcriptomic signatures enriched for ontology terms associated with growth regulation and apoptosis, inflammation, immune signaling, and autophagy in histologically normal prostate tissues and adjacent normal pancreatic cancer tissues that were identified as high-risk by DEGAS. The regions highlighted by DEGAS could reflect transcriptional precursors to intraepithelial neoplasia–a well-recognized premalignant morphological change in glandular epithelium. DISCUSSION/SIGNIFICANCE: Identifying biomarkers of tissue stress that precede morphologic diagnosis of high-grade pre-malignant lesions by a pathologist may help triage patients at high risk for future development of cancer, or aid in better understanding whether histologically normal pre-malignant tissues at tumor margins contribute to recurrence.
271 Diagnosis and Detection of Thrombosis in PCOS
Authors: Miller Mariel , Raymond Caitlin , Kavuri Bharathi , Zahner Christopher
Published: April 2023

OBJECTIVES/GOALS: Identify and stratify clinical presentations of thrombotic pathology in PCOS patients. This will be accomplished by 1) evaluating clinical assays for the detection of hypofibrinolysis, and 2) analyzing clinical symptomology of thrombosis in PCOS via Symptom-Disease Pair Analysis of Diagnostic Error (SPADE). METHODS/STUDY POPULATION: Preliminary study populations include n=3 for each of the following groups PCOS with thrombotic complications, PCOS without thrombotic complications, healthy controls, and healthy control samples treated ex vivo with PAI-1. Coagulation assays include ACL coagulation panel, antiphospholipid antibody assays, viscoelastic testing with TEG and Quantra devices, and global fibrinolytic capacity. Coagulation assays will be performed on three samples taken at 4-week intervals. The SPADE techniques will be used to evaluate symptomology of thrombosis in the study period and patient electronic medical history. Molecular testing will be performed for pro-thrombotic polymorphisms (PAI-1 and Apo E) and mutations (Factor V Leiden and Prothrombin). RESULTS/ANTICIPATED RESULTS: We anticipate PCOS and ex vivo PAI-1 samples to show signs of hypofibrinolysis on TEG and Quantra devices outside of reference ranges and with statistical significance. We also anticipate seeing a statistical significance with ACL coagulation panels, however, these results are expected to still be within the reference ranges, as seen in previous studies. We believe the SPADE method will identify clinical presentations of hypofibrinolysis in PCOS patients coinciding with patterns in laboratory tests that were misdiagnosed due to being within reference ranges. We hope to stratify clinical presentations predictive of thrombosis in PCOS patients with standard clinical assays and with increased precision using viscoelastic assays. DISCUSSION/SIGNIFICANCE: The incidence rate of thrombosis is 40x higher in PCOS compared to healthy populations. However, the mechanisms of thrombosis in PCOS are unknown and are undetectable with current clinical assays. We hypothesize that chronic cellular stress in PCOS disrupts the regulation of interconnected immune pathways, causing hypofibrinolysis.
274 Early life antibiotic exposure and genetic risk in neurodevelopmental disorders: effects on neurogenesis, the gut microbiome, and behavior
Authors: McDermott Courtney R. , Mirmaglesi Anya , Gao Zhan , Kimbark Katherine , Ntim Christiana , Zhang Xuesong , Zhou Xiaofeng , Millonig James H. , DiCicco-Bloom Emanuel
Published: April 2023

OBJECTIVES/GOALS: Our long-term goal is to understand how both genetic and environmental (GxE) factors contribute to neurodevelopmental disorders (NDDs) so that we may potentially intervene in disease pathogenesis and design therapies to address functional deficiencies. METHODS/STUDY POPULATION: Our studies use a novel GxE model to determine how cephalosporin antibiotic exposure alters the gut microbiome, hippocampal neurogenesis, and behavior in the genetically vulnerable 16p11.2 microdeletion (16pDel) mouse. This mouse models one of the most frequently observed genetic risk variants implicated in NDDs, including ~1% of autism diagnoses. Wildtype and 16pDel littermates were exposed to saline or the cephalosporin, cefdinir, from postnatal days 5-9. We quantified changes in gut microbiota composition using 16S rRNA gene sequencing and utilized immunoblotting, immunohistochemistry, and bulk RNA gene sequencing to assess changes in hippocampal neurogenesis. An additional cohort of saline or cefdinir-exposed mice were subjected to a behavioral battery to assess changes in sociability and anxiety. RESULTS/ANTICIPATED RESULTS: We leveraged the next-generation microbiome bioinformatics platform, Quantitative Insights Into Microbial Ecology 2 (QIIME2) to analyze 16S rRNA gene sequencing datasets of P13 cecal samples from saline- and cefdinir-exposed mice. We found successful perturbations to the gut microbiome following early life cefdinir exposure. Further, we found a robust 50% reduction in hippocampal cyclin E protein in cefdinir-exposed 16pDel male mice, which was replicated in a second independent experiment. This reduction extended to the S-phase cell entry and general stem cell population, quantified by EdU+ and Ki67+ cell numbers, respectively. Lastly, in our first cohort of mice for behavioral studies, we found reduced sociability and increased anxiety-like behaviors in cefdinir-exposed mice. DISCUSSION/SIGNIFICANCE: The findings from this GxE model will provide mechanistic insights into the causes of NDDs; they may inform practice guidelines so as to reduce this environmental exposure; and may suggest interventions like probiotics for those at risk in order to overcome altered gut microbiome composition and restore hippocampal neurogenesis defects.
283 Gait: a biomarker for pain and function in chronic low back pain
Authors: Bailes Anna H , Johnson Marit E. , Redfern Mark , Perera Subashan , Greco Carol , Sowa Gwendolyn , Cham Rakie , Bell Kevin
Published: April 2023

OBJECTIVES/GOALS: Chronic low back pain (cLBP) is associated with gait impairments. Gait may serve as an important biomarker for improvement following therapy interventions; however, gait has not been sufficiently studied in relationship to pain and function in cLBP. METHODS/STUDY POPULATION: Adults with cLBP completed a two-minute-walk-test around a 37.5 m oval track while wearing an inertial measurement unit (IMUâ€”Lifeware LLC, Pittsburgh, PA) over the L5 spinous process. Step time average, step time variability, step length, and symmetry (harmonic ratio) were calculated based on linear trunk accelerations, and gait speed was calculated based on distance walked. Participants completed the PEG tool (Pain, Enjoyment, General activity; scores closer to 10 indicate worse pain) to quantify pain intensity/interference and the PROMIS Physical Function SF-6b tool (mean t-scores=50 +/- 10; higher scores indicate better function) to quantify physical function. Pearson correlation coefficient (r) was used to determine strength of associations between gait and pain/physical function. RESULTS/ANTICIPATED RESULTS: Eleven adults (8 female, age 40 +/- 17, pain duration≥3 months) with cLPB participated in this study after completing an informed consent process approved by the University of Pittsburgh Institutional Review Board. Participants with a history of cancer, spinal cord compression, discitis, or activity restrictions prohibiting them from protocol completion were excluded. The mean PEG scores and PROMIS Physical Function t-scores were 2.8 +/- 1.8 and 47.8 +/- 8.2 respectively. There was a moderate-strong correlation between step time average and PEG (r=0.67, p=0.02), and a moderate-strong correlation between gait speed and PROMIS Physical Function (r=0.62, p=0.04). There were no other significant associations. DISCUSSION/SIGNIFICANCE: Gait speed and step time may be important movement biomarkers to consider when evaluating patients with cLBP. Generalizability of results are limited by the small study cohort and this cohort's relatively low pain burden and high level of physical function.
303 Investigation of the antibacterial and regenerative properties of a novel AHA dental coating for the treatment of deep caries
Authors: Asby Sarah , Binne Erin , Kehe Gannon , Kadur Pinky , Dharmala Devika , Schurr Michael , Puranik Chaitanya , Nair Devatha P.
Published: April 2023

OBJECTIVES/GOALS: Our objective is to investigate the antibacterial and regenerative properties of a novel AHA dental coating for the prevention and treatment of deep caries (cavities). Further, we aim to investigate and compare these properties through in vivo murine models and assessment on human saliva samples and human pulpal cells collected from clinical samples. METHODS/STUDY POPULATION: In vitro antibacterial studies were performed by collecting and culturing human salivary bacteria with AHA substrates and quantifying survival of cariogenic Sm (S. mutans). In vivo, C57BL/6 mice were treated with AHA composite fillings, infected with Sm clinical isolates, and fed a high sucrose diet with cavity formation assessment after 6 weeks. To evaluate regeneration, mice were similarly given composite with AHA or MTA (standard of care) upon pulpal exposure with regeneration quantified by microCT and histological analysis of dentin bridge formation, ALP production, and odontoblast migration. In vitro, AHA substrates were cultured with MC3T3-E1 pre-osteoblast cells and dental pulp stem cells obtained from clinical samples over 21 days, with mineralization and ALP assessed indicating osteogenesis. RESULTS/ANTICIPATED RESULTS: In vivo studies have shown the reduction of cavity formation in mice treated with AHA as well as dentin regeneration upon pulpal exposure using microCT and histological image analysis. Coinciding with these findings, AHA substrates eradicated cariogenic Sm in human saliva samples and single species cultures in vitro. Further, preliminary results in vitro have shown increased mineralization of MC3T3-E1 cells when cultured with AHA substrates in comparison to uncoated substrates. We anticipate similar increased mineralization as well as increased ALP production of human pulpal stem cells from clinical samples when cultured with AHA substrates, suggesting osteogenesis. Further, we anticipate increased odontoblast migration and ALP production upon additional analysis of in vivo tissue samples. DISCUSSION/SIGNIFICANCE: This work will elucidate the antibacterial and regenerative properties of AHA dental coatings. These results further support the translation of AHA coatings into the clinic as a novel therapeutic for the prevention and treatment of dental decay, which may overcome the limitations associated with the current treatments.
310 Mechanisms of progression of myelodysplastic syndrome to fatal secondary acute myeloid leukemia
Authors: Chang Daniel , Noble-Orcutt Klara , Lue Antony Marie , Largaespada David , Myers Chad , Sachs Zohar
Published: April 2023

OBJECTIVES/GOALS: Aim 1: Define the genetic events that cooperate with Trp53 mutations to mediate the transformation of MDS to sAML at the stem cell level. Aim 2: Use Bayesian networks to model the signaling pathway activation states identify aberrant regulators of self-renewal in LSCs. METHODS/STUDY POPULATION: To model the transformation of MDS, I will utilize a mouse model of MDS and sAML. The Trp53 mutated mouse mimics the genetics of human MDS and develops MDS. To discover how additional mutations contribute to disease progression, I use the Sleeping Beauty transposon system which induces random mutations. I propose to use this mouse model to define the molecular mechanisms of transformation of MDS to sAML. I will also measure levels of activated signaling intermediates in sAML using CyTOF. CyTOF is a method of flow cytometry that measures up to 40 proteins simultaneously at single-cell resolution. I will also use Bayesian network to model the signaling architecture of the MDS/sAML stem cells. RESULTS/ANTICIPATED RESULTS: I will identify putative mediators of self-renewal in gene expression data and the corresponding Bayesian networks model. I will prioritize genes and signaling intermediates that act in pathways that rank highly in both methods and those that have previously published roles in self-renewal. Based on prior work, I will focus on EZH2, NFï «B, and mTOR pathways in addition to candidate pathways revealed by these studies. Small molecule inhibitors that target candidate signaling intermediates will be used for experimental validation. I will test the impact of these small molecule inhibitors on LSC self-renewal using serial colony forming assays as well as in vivo mouse reconstitution assays. DISCUSSION/SIGNIFICANCE: We focus on the role of stem cells in the transformation of MDS to sAML. Understanding the mutations and signaling relationships that mediate self-renewal in Trp53 mutant stem cells will allow for precise therapeutic target of this disease and improve outcomes for these patients.
312 Metabolomics Approach to Cellular Senescence: Characterizing the Secretory Phenotype and Metabolism of Irradiation-Induced Senescent Human Pre- Adipocytes
Authors: Ortiz-Correa Zinnarky Kiani , Lanza Ian R.
Published: April 2023

OBJECTIVES/GOALS: The study aims to identify and measure metabolites in irradiation-induced senescent pre-adipocytes induced by the increased secretion of pro-inflammatory cytokines. Through untargeted metabolomics, we seek to understand the alterations to metabolism and mitochondrial activity that occur during irradiation-induced senescence. METHODS/STUDY POPULATION: First, commercially available human primary subcutaneous pre-adipocytes were cultured in vitro, and irradiated to induce senescence. To confirm senescence, cells were stained for beta-galactosidase, which was positive in senescent pre-adipocytes. The cells and their conditioned cultured media were then collected and frozen for untargeted metabolomics to profile metabolites. The sample analysis is currently underway and will be conducted using central carbon isotope tracing and chromatograph mass spectrometry. Principal Component Analysis, fold change analysis, and heat maps will be used to detect and report the changes in metabolite signals. Oxygen consumption rate and extracellular acidification rate measurements are in progress at present using the Agilent Seahorse XF Cell Mito Stress Test. RESULTS/ANTICIPATED RESULTS: We expect that metabolites of central carbon metabolism and oxidative phosphorylation will be upregulated. The concentrations of metabolites of pathways altered by the pro-inflammatory cytokines that have been identified to be secreted by senescent cells are expected to be altered. The metabolites measured in conditioned culture media will provide insight into the changes to the cellular microenvironment caused by senescence. Finally, we anticipate that mitochondrial function, and both aerobic and anaerobic metabolism will be altered in senescent pre-adipocytes compared to controls. Through the present study, we will achieve a better understanding the metabolic alterations that occur as part of cell senescence in pre-adipocytes. DISCUSSION/SIGNIFICANCE: Adipose tissue dysfunction due to the accumulation of senescent cells has been linked to chronic diseases such as diabetes and insulin resistance, and inflammaging. Through this study, we expect to provide new insight into the metabolic alterations of senescence and offer a backbone for prospective translational human studies and clinical trials.
335 Sources of Sound Exposure in Pediatric Critical Care
Authors: Kalvas Laura Beth , Harrison Tondi M.
Published: April 2023

OBJECTIVES/GOALS: Sleep is critical for healing, however pediatric intensive care unit (PICU) sound is above recommended levels (i.e., 45 A-weighted decibels [dBA]). This observational study identifies sources of PICU sound and compares sources between times of high (i.e., dBA≥45) and low (i.e., dBA < 45) levels. METHODS/STUDY POPULATION: The sound environment of 10 critically ill children 1 to 4 years of age was monitored via a bedside dosimeter and video camera for 48 hours, or until PICU discharge. Dosimeter and video data were uploaded to Noldus Observer XT and time synchronized. A reliable, previously published coding scheme developed to identify sound sources in the adult ICU was modified for the pediatric population. Sound sources (e.g., clinician/family/child [verbal vs. non-verbal] vocalization, patient care, medical equipment) were identified via instantaneous sampling of video data at each minute of recording. The proportion of sampling points with each sound source are compared between times of high and low sound levels, and between day (7:00-18:59) and night (19:00-6:59) shift. RESULTS/ANTICIPATED RESULTS: Video coding is ongoing, with high inter-rater reliability (ÎºÌ…=0.99, SD DISCUSSION/SIGNIFICANCE: Medical equipment sound is ubiquitous in the PICU. Clinicians should optimize the PICU sound environment for sleep, including minimizing equipment alarms, conversation, general activity, and screen media during child rest. Large-scale studies are needed to confirm findings from this small cohort.
338 The Alabama Genomic Health Initiative: Integrating Genomic Medicine into Primary Care
Authors: Limdi Nita A , Absher Devin , Asif Irf , Bateman Lori , Barsh Greg , Bowling Kevin M. , Cooper Gregory M. , Davis Brittney H. , East Kelly M. , Finnila Candice R. , Goff Blake , Hiatt Susan , Kelly Melissa , Kelley Whitley V. , Korf Bruce R. , Latner Donald R. , Lawlor James , May Thomas , Might Matt , Moss Irene P. , Nakano-Okuno Mariko , Osborne Tiffany , Sodeke Stephen , Stout Adriana , Thompson Michelle L.
Published: April 2023

OBJECTIVES/GOALS: Supported by the State of Alabama, the Alabama Genomic Health Initiative (AGHI) is aimed at preventing and treating common conditions with a genetic basis. This joint UAB Medicine-HudsonAlpha Institute for Biotechnology effort provides genomic testing, interpretation, and counseling free of charge to residents in each of Alabama’s 67 counties. METHODS/STUDY POPULATION: Launched in 2017, as a state-wide population cohort, AGHI (1.0) enrolled 6,331 Alabamians and returned individual risk of disease(s) related to the ACMG SF v2.0 medically actionable genes. In 2021, the cohort was expanded to include a primary care cohort. AGHI (2.0) has enrolled 750 primary care patients, returning individual risk of disease(s) related to the ACMG SF v3.1 gene list and pre-emptive pharmacogenetics (PGx) to guide medication therapy. Genotyping is done on the Illumina Global Diversity Array with Sanger sequencing to confirm likely pathogenic / pathogenic variants in medically actionable genes and CYP2D6 copy number variants using Taqman assays, resulting in a CLIA-grade report. Disease risk results are returned by genetic counselors and Pharmacogenetics results are returned by Pharmacists. RESULTS/ANTICIPATED RESULTS: We have engaged a statewide community (>7000 participants), returning 94 disease risk genetic reports and 500 PGx reports. Disease risk reports include increased predisposition to cancers (n=38), cardiac diseases (n=33), metabolic (n=12), other (n=11). 100% of participants harbor an actionable PGx variant, 70% are on medication with PGx guidance, 48% harbor PGx variants and are taking medications affected. In 10% of participants, pharmacists sent an active alert to the provider to consider/ recommend alternative medication. Most commonly impacted medications included antidepressants, NSAIDS, proton-pump inhibitors and tramadol. To enable the EMR integration of genomic information, we have developed an automated transfer of reports into the EMR with Genetics Reports and PGx reports viewable in Cerner. DISCUSSION/SIGNIFICANCE: We share our experience on pre-emptive implementation of genetic risk and pharmacogenetic actionability at a population and clinic level. Both patients and providers are actively engaged, providing feedback to refine the return of results. Real time alerts with guidance at the time of prescription are needed to ensure future actionability and value.
347 The tradeoff between kinematic and muscular control of reaching as a potential biomarker of motor performance in stroke
Authors: Brunfeldt Alexander T , Bregman Barbara S , Lum Peter S
Published: April 2023

OBJECTIVES/GOALS: Nearly 3 million Americans live with arm impairment following stroke. While as many as 20% of patients fully recover, individual differences in recovery make one-size-fits-all rehabilitation approaches suboptimal. The goal of this study was to use our custom rehabilitation platform to identify neuromuscular biomarkers of arm control in stroke. METHODS/STUDY POPULATION: Chronic stroke survivors (N = 10) reached for targets in a virtual reality environment using both hands. They completed 162 reaches divided into 3 blocks. Following baseline, we used our custom exoskeletons to provide 50% arm weight assistance to the impaired limb and 50% arm weight resistance to the non-impaired limb. We removed the exoskeletons during the retention block. We used electromyography to approximate muscle activity in the anterior deltoids. Relative contribution (RC) was calculated as the displacement of the impaired arm divided by the sum of displacements for both arms. Muscle contribution (MC) was calculated as the root mean square of impaired arm muscle activity divided by the sum of activity for both deltoids, normalized to maximum voluntary contraction. RESULTS/ANTICIPATED RESULTS: During baseline, RC of the impaired limb was 43%; patients reached significantly less with their impaired arm compared to their non-impaired arm (p = 0.02). MC of the impaired deltoid was 56% and was similar between arms (p = 0.5). During loading, RC did not change relative to baseline (p = 0.87), but MC tended to decrease by 11% (p = 0.12). These results suggest a tradeoff between kinematic and muscular control of reaching. This new finding closely matches our previous work in 12 healthy controls, where we found a 2% increase in RC and a 11% decrease in MC. Importantly, 4/10 patients exhibited an inverse tradeoff (i.e., decrease in RC and/or increase in MC). We will analyze neuroimaging data to determine the role lesion size and location play in predicting an individual’s response to gravity compensation. DISCUSSION/SIGNIFICANCE: Our tradeoff analysis serves as a potential neuromuscular biomarker of stroke survivors’ responsiveness to gravity compensation. This forms the basis for personalized technologies for stroke rehabilitation. With further development, clinicians can use our platform to fine-tune compensation levels based on the individual needs of the patient.
354 Unitary neural correlates of executive control in pediatric transdiagnostic psychopathology
Authors: Kaminski Adam , Xie Hua , You Xiaozhen , Flaharty Kathryn , Jeppsen Charlotte , Li Sufang , Merchant Junaid S. , Berl Madison M. , Kenworthy Lauren , Vaidya Chandan J.
Published: April 2023

OBJECTIVES/GOALS: Childhood psychiatric symptoms are highly comorbid. Their co-occurrence and association with negative life outcomes is partially explained by deficits in executive control, or processes enabling self-regulation. Here, we test a novel executive neural target in three fMRI tasks and its relevance to shared psychopathology. METHODS/STUDY POPULATION: We studied 60 children [15 F/45 M; mean age (SD)=11.6 years (1.62)] with diverse diagnoses including attention deficit disorder (n=26) and autism spectrum disorder (n=22). We extracted a latent general factor of psychopathology using principal component analyses applied to parent-report Child Behavior Checklist syndrome scores. Subjects completed 3 executive control fMRI probes, tapping adaptive control, working memory, and inhibition. Correlational psychophysiological interaction (cPPI) analysis measured correlations between executive control-related modulations of activity in 414 network-affiliated parcels. We selected parcels exhibiting control-related cross-network correlations as well as control-related activity across all tasks and tested them for association with psychopathology. RESULTS/ANTICIPATED RESULTS: cPPI connectivity matrices were thresholded and graphs were identified using the Network-Based Statistic toolbox (p90th percentile PC) as well as control-related activation (>10% activated voxels; p DISCUSSION/SIGNIFICANCE: Our results examine cross-network interactions between brain regions during 3 fMRI tasks and their role in explaining individual variation in psychopathology. As executive control links to both comorbidity and life outcomes, identifying the clinically-relevant neural correlates of controlled behavior may lead to transdiagnostic treatments.
357 Using Assessments to Create a Translational Pipeline at a Science-Based Inpatient Addiction Treatment Facility
Authors: Bourdon Jessica , Fields Taylor , Judson Sidney , Vadhan Nehal , Morgenstern Jon
Published: April 2023

OBJECTIVES/GOALS: Effective translation of data to inform real-time patient care is lacking in addiction inpatient settings. The current study presents the optimization of a Comprehensive Rehabilitation Assessment summary report that is used by clinicians to individualize treatment. METHODS/STUDY POPULATION: A multi-aim approach was taken that utilized aspects of various implementation science frameworks. Participants were clinical staff (N = 7; female = 71%). A quantitative survey was used for aims 1 and 2 to assess motives and context around the report as well as evaluate the design of it. Aim 3 focused on optimization via semi-structured interviews. Descriptive and modified content analyses were utilized appropriately for each aim. RESULTS/ANTICIPATED RESULTS: Five versions of the assessment report were created between February 2021 and August 2022, the most recent of was adapted into patients’electronic medical records based on study results. Each report version, participants’results/feedback, and researchers’perceived barriers to this translational process will be discussed. DISCUSSION/SIGNIFICANCE: The current study highlights a replicable approach for optimizing the translation of assessment data into treatment for patients with disorders of addiction.
372 HMGB1 Localization as a Driver of Carcinogenesis in RDEB-Associated Squamous Cell Carcinoma
Authors: Bui Kacey Guenther , Bielinsky Anja , Nguyen Hai Dang , Tolar Jakub
Published: April 2023

OBJECTIVES/GOALS: This study investigates whether localization of high mobility group box 1 (HMGB1) controls inflammatory signaling and DNA damage response in human keratinocytes, the cell of origin for squamous cell carcinoma (SCC). SCC is especially metastatic in chronic wounds, burns, and in patients with recessive dystrophic epidermolysis bullosa (RDEB). METHODS/STUDY POPULATION: We used CRISPR/Cas9 gene-editing to knock out HMGB1 in a keratinocyte line, p16INK4a-negative keratinocytes immortalized by ectopic hTERT expression (N/TERT-2G [46,XY]). Following gene editing, clonal keratinocyte populations were screened for knockout by PCR followed by TIDE analysis (Tracking of Indels by Decomposition) to identify indels that would result in a frameshift mutation. Total cell lysates for each clonal population were analyzed by immunoblot and immunofluorescence for HMGB1 protein. These cells will be used to assay for DNA damage sensitivity in the presence of genotoxic agents (etoposide, ultraviolet radiation, γ-irradiation). A lentiviral vector will then be used to express mutant forms of HMGB1 that localize to the nucleus (C23/45S) or cytoplasm (C106S) and DNA damage assays repeated. RESULTS/ANTICIPATED RESULTS: We have confirmed by sequencing, immunoblot, and immunofluorescence that HMGB1 is knocked out in a clonal population of N/TERT-2G human keratinocyte cells. We anticipate that cells with a complete absence of HMGB1 will have high sensitivity to DNA damaging agents, but little change in inflammatory signaling. We also expect that cells expressing mutant HMGB1 that localizes exclusively to the cytoplasm will demonstrate an increased sensitivity to DNA damage relative to wild-type controls, while mutant HMGB1 that localizes exclusively to the nucleus will be protected from DNA damage caused by exposure to genotoxic agents. DISCUSSION/SIGNIFICANCE: HMGB1 is a nuclear protein and damage associated molecular pattern that is elevated systemically in patients with RDEB, many of whom will go on to develop metastatic squamous cell carcinoma. The experiments described here investigate whether HMGB1 plays a mechanistic role in skin carcinogenesis via regulation of the DNA damage response.
383 A Pilot Study of Tear Cytokine Profiling in a Patient with Ocular Graft versus Host Disease
Authors: Sunshine Sarah B. , Li Andrew , Gausch Fernando Martinez , Cao Xuefang , Atanackovic Djordje
Published: April 2023

OBJECTIVES/GOALS: Ocular graft versus host disease (oGVHD) affects ~50% of individuals after an allogeneic hematopoietic stem cell transplant for treating blood cancers. OGVHD results in severe dry eye disease and decreased vision. Quantifying specific cytokine changes in tears may reveal biomarkers and future treatment targets for patients with oGVHD. METHODS/STUDY POPULATION: The goal of this study is to determine if cytokines can be measured in tears with the Isoplexis platform. This pilot validation study evaluates the tears of a patient with oGVHD utilizing the Isoplexis platform. The Isoplexis has specific advantages for tear samples including high-throughput analysis, and small sample requirements, but has yet to be validated in tears. A sample from normal and oGVHD patient tears were collected for comparison. Samples were analyzed on two separate backgrounds–standard Bovine Serum Albumin (BSA) background and artificial tears (ATs). The negative control was ATs and positive control was a concentrated cytokine solution. Analysis of 22 cytokines was performed. RESULTS/ANTICIPATED RESULTS: Analysis of 22 cytokines was performed. As expected, the cytokine levels of the ATs alone were below the limit of detection (LOD). The oGVHD patient tears showed elevated TNF-alpha, TNF-beta, perforin, MIP-1a, MIP-1β, MCP-1, IL2, IL4, IL5, IL-7A, IL9, IL-13, IL-15, IFN-γ, granzyme B, and GM-CSF with ATS background, but no cytokines above the LOD in the BSA background plate. The control tears had elevated IP-10. The elevated cytokines for the oGVHD patient corresponded to symptom severity and clinical findings. DISCUSSION/SIGNIFICANCE: These results suggest that using ATs as the background with the Isoplexis platform improves the sensitivity to detect tear cytokines. Findings of elevated IL-7A and GM-CSF in tears parallels literature findings for oGVHD. Further evaluation of samples will continue to validate the Isoplexis multiplex assay for tear cytokine analyses.
392 Basic Researcher Interviews to iDentify Gaps to Enabling Translation (BRIDGE Translation)
Authors: Thepmankorn Parisorn , Tafuto Barbara , Gonzalez Anthony , Anwar Farah , Gelinas CÃ©line , Fiedler Nancy
Published: April 2023

OBJECTIVES/GOALS: Despite expanded interest in translational research, barriers in funding, infrastructure, staffing, training opportunities, and interdisciplinary collaboration still remain. Our goal was to interview basic science researchers to identify research barriers and potential areas for improvement. METHODS/STUDY POPULATION: After receiving an IRB determination for a quality improvement study, 15 New Jersey-based principal investigators (PIs) from various departments and at various stages of their careers were virtually interviewed one-on-one by a trained medical student and asked a series of standardized questions about their subjective experiences with their institutions’ research processes, training and mentoring, interdisciplinary collaboration, and intellectual property. The interview was then transcribed to complete an anonymous, standardized REDCap form. Qualitative data analysis was performed to identify common themes, barriers, and gaps in conducting translational research as reported by the PIs. RESULTS/ANTICIPATED RESULTS: Of the 15 PIs, 4 (27%) were assistant professors, 6 (40%) associate professors, and 5 (33%) professors. 5 (33%) joined the institution less than 5 years ago. The most common barrier was translational research funding. Time needed to navigate administrative and regulatory processes and access to clinical collaborators were other common barriers. One PI noted leaving the institution due to difficulty accessing clinicians and patient samples. PIs with extensive training or who reported successfully conducting translational research noted fewer barriers. Suggested solutions included programs and grants to link basic scientists with clinicians, a streamlined IRB process, and better staffing to support research. DISCUSSION/SIGNIFICANCE: The findings suggest a need to increase grant funding for translational research. Improving support staffing and minimizing administrative barriers would also be helpful. Improving the dissemination of available resources, grants, and guidance on administrative processes may further decrease barriers.