bluuebunny/crossref_metadata_2025 huggingface dataset:
4396 Immunoglobulin administration and hypogammaglobulinemia during pediatric acute leukemia therapy
Authors: Edington Holly , Chandrakasan Shanmuganathan , Miller Tamara , DeGroote Nicholas , Mertens Ann , Castellino Sharon
Published: July 2020

OBJECTIVES/GOALS: Intravenous immunoglobulin (IVIG) is used for infection prevention in pediatric B-cell acute lymphoblastic leukemia (B-ALL), but evidence for this is lacking. We describe the prevalence of hypogammaglobulinemia in pediatric B-ALL, predictors of IVIG use and its efficacy for infection prevention. METHODS/STUDY POPULATION: We will conduct a retrospective review of children age 1-21 years with B-ALL treated at Aflac Cancer and Blood Disorders Center from 2010 to 2017. The cohort was identified through the cancer registry. Demographics, disease factors, laboratory values, medications and infection outcomes were linked between the electronic medical record and an institutional database. Outcomes of interest include emergency department (ED) visits, hospitalization days, and episodes of infection. Descriptive statistics will be performed. Outcomes will be compared between IVIG recipients and non-recipients. Univariate and multivariate logistic regression models will assess predictors of IVIG administration. RESULTS/ANTICIPATED RESULTS: We identified 443 patients with B-ALL during the study period who met inclusion criteria. Exclusion criteria included receipt of IVIG or hematopoietic stem cell transplant prior to diagnosis. The average age at diagnosis is 6.5 years (standard deviation 4.8 years); 52.6% are male; 61.6% are white; 61.0% are standard risk per National Cancer Institute criteria. Among eligible patients, 137 (31.1%) received IVIG. We hypothesize that IVIG initiation is associated with hypogammaglobulinemia and history of severe infection. We also anticipate that frequency of emergency department visits, hospitalization days, and episodes of infection will decrease after IVIG initiation. DISCUSSION/SIGNIFICANCE OF IMPACT: The immunological profile of children with B-ALL and factors influencing their susceptibility to infection are still incompletely understood. The benefits of IVIG are unknown. This study will provide evidence for IVIG prophylaxis recommendations in pediatric leukemia patients.
4077 Listening with the HEAR-QL: Quality of Life in Children with Hearing Loss
Authors: Wahba Brandon Malik , Lieu Judith
Published: July 2020

OBJECTIVES/GOALS: This study evaluates the utility of self-reported quality of life measure in children with hearing loss. We will compare self-reported HEAR-QL scores with parent-reported HEAR-QL scores. We will then test the relationship between HEAR-QL scores and scores on a standardized assessment of cognition, the NIH Cognition Battery. METHODS/STUDY POPULATION: We will administer the HEAR-QL questionnaire to children with hearing loss and their parents. We will then administer the NIH Cognition Battery to the child. We will include in our population children ages 7 to 14 with hearing loss of any severity or side. We will exclude those with intellectual disability, disorders of speech or language, or those who would be unable to complete the questionnaires for any reason. Children will be recruited from Otolaryngology clinics at St. Louis Children’s Hospital based on ICD diagnosis of sensorineural hearing loss between 01/2015 – 03/2020. RESULTS/ANTICIPATED RESULTS: We will aim to recruit 44 patients in total, which is the sample size needed to detect a moderate correlation (r = 0.4) with a 1-sided α = 0.05 and 1-β = 0.8. HEAR-QL scores and NIH Cognition Battery scores will be reported using descriptive statistics. Linear regression as well as correlation analysis between HEAR-QL scores and cognitive testing scores will be performed using a 1-sided α = 0.05, with 1-β = 0.8. If recruitment is sufficient, we will adjust for demographics that are significantly correlated with the outcome on multivariate analysis. Finally, we will test for agreement between parent report and child report by calculating a Kappa statistic. DISCUSSION/SIGNIFICANCE OF IMPACT: There is little clarity on the necessity of amplification in children with hearing loss, yet the child’s perspective is not routinely assessed in clinical practice. This study employs self-report in a pediatric population with hearing loss to find out if children provide new and reliable information.
4379 MARCKS protein is altered in naturally occurring model of asthma in horses
Authors: Davis Kaori , Sheats Mary
Published: July 2020

OBJECTIVES/GOALS: Asthma is a significant health concern that affects people of all ages worldwide. EAS demonstrates many of the pathophysiological characteristics of nonatopic human asthma, which has led EAS to be used as naturally occurring model. Previous work from our lab determined that MARCKS (Myristoylated Alanine Rich C Kinase Substrate) protein is an essential regulator of cellular inflammatory functions. In the current study, we hypothesized that MARCKS levels would be increased in BAL cell lysates from horses with EAS, and that inhibition of MARCKS in zymosan-stimulated BAL cells (ex vivo) would diminish respiratory burst. METHODS/STUDY POPULATION: Lysates were prepared from BAL cells isolated from horses with no, mild/moderate and severe EAS. Relative MARCKS protein levels were determined using equine specific MARCKS ELISA (MyBioSource). Cultured BAL cells were pretreated with a MARCKS inhibitor peptide (MANS), control peptide (RNS) or vehicle control and stimulated with zymosan for 5 hours. Reactive oxygen species levels were determined by luminescence to evaluate respiratory burst. Data were analyzed by One-way ANOVA (p<0.05). RESULTS/ANTICIPATED RESULTS: We determined that normalized MARCKS protein expression is significantly increased in BAL cell lysates from horses with mild/moderate or severe EAS, compared to horses with normal BAL cytology. Preliminary findings also suggest that MANS treatment of zymosan-stimulated equine BAL cells ex vivo attenuates levels respiratory burst. DISCUSSION/SIGNIFICANCE OF IMPACT: These findings point to a possible role for MARCKS protein in the pathophysiology of EAS and support MARCKS inhibition as a potential therapeutic strategy.
4377 Missed Opportunities to Prevent Homicide: An Analysis of the National Violent Death Reporting System
Authors: Cirone Justin , Cone Jennifer , Williams Brian , Hampton David , Prakash Priya , Zakrison Tanya
Published: July 2020

OBJECTIVES/GOALS: The goal of this study is to better understand the homicide victim population who were institutionalized within 30 days prior to their death. Improved knowledge of this population can potentially prevent these future homicides. METHODS/STUDY POPULATION: A retrospective analysis of the 36 states included in the 2003-2017 National Violent Death Reporting System was performed. Demographics of recently institutionalized homicide victims (RIHV) in the last 30 days were compared to homicide victims who were not recently institutionalized. Circumstances of the homicide, such as suspected gang involvement, were also compared. Parametric and non-parametric statistical analyses were performed. Significance was set at p<0.05. RESULTS/ANTICIPATED RESULTS: There were 81,229 homicides with 992 (1.2%) RIHV. The majority of RIHV were Black (49.6%) and older than victims who were not recently institutionalized (37.2 vs. 34.8, p<0.001). RIHV had a high school degree or higher in 54.8% of cases and the primary homicide weapon was a firearm in 67% of the deaths. They were more likely to be homeless (3.1% vs. 1.5%), have a mental health diagnosis (9.2% vs. 2.3%), abuse alcohol (6.1% vs. 2.2%), or abuse other substances (15.2% vs. 5.8%) [all p <0.001]. These victims were most commonly institutionalized in a correctional facility or a hospital compared to other facilities such as nursing homes. Homicide circumstances for RIHV were more likely to involve abuse/neglect (4.3% vs. 2.2%, p<0.001), gang violence (7.6% vs. 5.6%, p = 0.002), or a hate crime (1.0% vs. 0.1%. p<0.001). DISCUSSION/SIGNIFICANCE OF IMPACT: Contact with an institution such as a hospital or prison provides high-risk patients the opportunity to potentially participate in violence intervention programs. These institutions should seek to identify and intervene on this population to reduce the risk of violent homicides.
4141 Molecular Signatures of Cocaine Toxicity in Postmortem Human Brain and Neurons
Authors: Mendez Emily Frances , Stertz Laura , Fries Gabriel , Hu Ruifeng , Meyer Thomas , Zhao Zhongming , Walss-Bass Consuelo
Published: July 2020

OBJECTIVES/GOALS: The goal of this project is to identify new therapeutic targets and biomarkers to treat or prevent cocaine toxicity by investigating proteomic, transcriptomic and epigenetic signatures of cocaine exposure in human subjects. METHODS/STUDY POPULATION: Cocaine is a highly addictive neurotoxic substance, and it is estimated that 1.9 million Americans are current users of cocaine. To study the molecular effects of cocaine, we generated preliminary proteomics and next-generation RNA sequencing (RNAseq) data from human postmortem dorsolateral prefrontal cortex (Broadmann area 9 or BA9) of 12 cocaine-exposed subjects and 17 controls. Future directions for this project include RNAseq and DNA methylation analysis of neuronal nuclei sorted from human postmortem BA9 and a human induced pluripotent stem cell-derived neuron (hiPSN) model of cocaine exposure from the same postmortem subjects from whom we have brain samples. RESULTS/ANTICIPATED RESULTS: We found alterations in neuronal synaptic protein levels and gene expression, including the serotonin transporter SLC6A4, and synaptic proteins SNAP25, SYN2, SYNGR3. Pathway analysis of our results revealed alterations in specific pathways involved with neuronal function including voltage-gated calcium channels, and GABA receptor signaling. In the future, we expect to see an enhancement in neuron-specific gene expression signatures in our sorted neuronal nuclei and our hiPSN model of cocaine exposure. The hiPSN model will help elucidate which effects are due to acute versus chronic exposure of cocaine. DISCUSSION/SIGNIFICANCE OF IMPACT: Neuronal signatures found with this analysis can help us understand mechanisms of cognitive decline in long-term cocaine users as well as the acute effects on the brain of cocaine taken in overdose. With this work and future proposed studies, we can discover novel clinical biomarkers for cocaine neurotoxicity in patients with cocaine use disorder and determine readouts for future therapeutic development on cocaine addiction and overdose.
4226 Poor provider-patient communication, lack of readiness for discharge, and perceived illness threat are associated with quality of life after survival from cardiac arrest
Authors: Presciutti Alex , Shaffer Jonathan , Newman Mary , Perman Sarah
Published: July 2020

OBJECTIVES/GOALS: Studies have shown that cardiac arrest survivors have poor quality of life (QoL) secondary to neurologic injury. We hypothesized that poor provider-patient communication, lack of readiness for discharge, and perceived illness threat would be associated with QoL in cardiac arrest survivors. METHODS/STUDY POPULATION: We distributed an online survey to the Sudden Cardiac Arrest Foundation listserv. Survivors completed the Questionnaire for the Quality of Provider-Patient Interactions (QQPPI), Readiness for Hospital Discharge Scale (RHDS), and the Brief Illness Perception Questionnaire (B-IPQ). When completing the QQPPI and RHDS, survivors were asked to think back to their hospitalization and discharge. QoL domains (physical, psychological, social) were measured via the WHO-QOL BREF. Three multiple regression models examined associations between QQPPI, RHDS, and B-IPQ scores with QoL domains, adjusted for age, sex, months since arrest, and understanding of arrest and post-arrest symptoms at discharge. RESULTS/ANTICIPATED RESULTS: A total of 163 survivors (mean age 50.1 years, 50.3% women) provided complete survey data. Greater perceived illness threat (β: −.45, p < .001) and lower readiness for discharge (β: .22, p = .01) were associated with worse physical QoL; greater perceived illness threat (β: −.45, p < .001) was associated with worse psychological QoL; and greater perceived illness threat (β: −.3, p < .001) and poor provider-patient communication (β: .35, p < .001) were associated with worse social QoL. Our models explained 48%, 43%, and 30% of the variance in physical, psychological, and social QoL, respectively (p < .001). DISCUSSION/SIGNIFICANCE OF IMPACT: In-hospital interactions and perceived illness threat have important ramifications for cardiac arrest survivors attempting to return to daily life. Discussions regarding cardiac arrest sequelae, expectations, and specific treatment options during hospitalization could impact future QoL.
4336 Renal Tubular Complement C9 Deposition is Associated with Renal Tubular Damage and Fibrosis in Lupus Nephritis
Authors: Wang Shudan , Wu Ming , Chiriboga Luis , Putterman Chaim , Broder Anna , Belmont H. Michael
Published: July 2020

OBJECTIVES/GOALS: Tubulointerstitial damage in lupus nephritis (LN) is a strong predictor of progression to chronic kidney disease and end stage renal disease (ESRD). While complement activation mediates glomerular injury, the role of complement in renal tubular damage has not been evaluated. We investigated the association between complement activation and tubulointerstitial fibrosis. METHODS/STUDY POPULATION: Patients with LN were selected randomly between July 2014 - July 2016. Chromogenic immunohistochemistry was performed on formalin-fixed, paraffin-embedded, 4-µm human renal biopsy sections using unconjugated, murine anti-human Complement C9 (Hycult Biotech, clone X197) as a marker of the terminal complement activation. Positive control is C3 glomerulopathy and negative control is normal kidney. Tubular basement membrane C9 staining intensity were analyzed on semiquantitative scale 0 to 3 by a renal pathologist. Interstitial fibrosis/tubular atrophy were categorized into low (0–10%), medium (11–20%), or high (≥21%). Clinical parameters were assessed at time of biopsy and 6 months post biopsy. Bivariate associations were assessed between presence of tubular C9 (C9+) and other covariates. RESULTS/ANTICIPATED RESULTS: Renal biopsies from 30 LN studied, 23 (77%) of which had proliferative LN. There were 24 (80%) women, mean (SD) age 33 (12) years. Positive tubular C9 staining was observed in 7/30 (23%) biopsies. At time of renal biopsy, C9+ patients had significantly higher urine protein, compared to C9- patients: median (IQR) 6.2g (3.3-13.1) vs. 2.4g (1.3-4.6), p<0.01. The differences persisted at 6 months after induction therapy: 1.08g (1.0-8.3) in C9+ vs. 0.68g (0.2-2.1) in C9- patients, p = 0.06. There was no significant difference in creatinine at renal biopsy between the two groups. Tubular C9 deposition was associated with interstitial fibrosis: 49% had severe interstitial fibrosis vs. none in the C9- group, p = <0.01. Higher proportion of C9+ patients had moderate NIH Chronicity index: 42.9% vs 8.7% in the C9- group, p = 0.07. DISCUSSION/SIGNIFICANCE OF IMPACT: Tubular C9 deposition is significantly associated with proteinuria, interstitial fibrosis and increased chronicity which predict progression to ESRD and high mortality. This finding suggests that complement activation in the tubules may be linked to proteinuria and contribute to mechanism in tubulointerstitial damage in LN.
4444 The effect of early life antibiotics on gut microbiome and fecal bile acid concentrations in children
Authors: Benitez Alain Jesus , Gerber Jeffrey S. , Tanes Ceylan , Bittinger Kyle , Friedman Elliot S. , Li Hongzhe , Bushman Frederic D. , Wu Gary D. , Zemel Babette S.
Published: July 2020

OBJECTIVES/GOALS: The current proposal seeks to investigate the effect of early life antibiotic use in the development of functional gastrointestinal (GI) disorders. We propose that infants exposed to antibiotics will present with gut microbial dysbiosis, changes in fecal bile acid concentrations and develop more GI symptoms compared to unexposed children. METHODS/STUDY POPULATION: We analyzed fecal samples from 174 subjects at 12 months of age, of whom 52 were exposed to antibiotics in their first year of life. Of these, 33 subjects were sampled again at 24 months of age. DNA from 200mg of frozen stool (−80C) was isolated with the Qiagen DNeasy PowerSoil kit. Shotgun libraries were generated using the NexteraXT kit and sequenced on the Illumina HiSeq 2500 using 2x125 bp chemistry. Sequence data were analyzed using the Sunbeam metagenomics pipeline. The abundance of bacteria was estimated using Kraken version 2.0.8. Fecal bile acids will be quantified by liquid chromatography–mass spectrometry (LC-MS). RESULTS/ANTICIPATED RESULTS: Overall bacterial community composition at 12 or 24 months was not associated with antibiotic exposure (PERMANOVA test, Bray-Curtis distance). An increase in Enterobacteriaceae, in particular Escherichia coli, is a signature of antibiotic-induced dysbiosis, but also of early infant gut. Children with antibiotic exposure had slightly higher abundance of Escherichia coli compared to those with no exposure (p = 0.03). At 24 months, the abundance of Bacteroides caccae, a commensal gut species, was decreased for children exposed to antibiotics in the first year of life (fdr = 0.02). We will perform further analysis of bile acid modifying bacteria, fecal bile acid concentrations and correlate to GI symptoms. DISCUSSION/SIGNIFICANCE OF IMPACT: Our findings suggest a significant but nuanced impact of early life antibiotic use on the composition of the gut microbiota. The association of antibiotic exposure with B. caccae and E. coli warrant further attention in the context of the rapidly developing early-life microbiome. CONFLICT OF INTEREST DESCRIPTION: The authors declare no conflicts of interest relevant to this work.
4566 The Impact of Migration on Viral Hepatitis Prevalence and Elimination Among 30 European Nations: A modeling approach
Authors: Valles Kenneth , Inzunza Andres , Prasai Kritika , Roberts Lewis R
Published: July 2020

OBJECTIVES/GOALS: Hepatitis B and C virus causes inflammation of the liver and can lead to cirrhosis, liver failure, and hepatocellular carcinoma. The aim of this study is to generate a modeled estimate of changes in hepatitis B and C prevalence, and future sequelae, that accounts for recent mass migration to the European Union stemming from 50 high-emigration countries. METHODS/STUDY POPULATION: Total migrant population from 2013-2017 was obtained from the Eurostat population database. Demographics including country-of-origin, sex, and age distributions were used to determine migrant contributions to HBV and HCV prevalence where available. Undocumented migration estimates were obtained from the Institute of Migration database. Country-of-origin HBV and HCV prevalences were obtained for the select 50 country-of-origin nations from the Polaris Observatory and from systematic reviews. Disease progression was estimated using HBV and HCV outcome data for total populations from treatment guideline publication from the European Association for the Study of the Liver. RESULTS/ANTICIPATED RESULTS: Between 2013 and 2017, a total of 11,030,786 documented migrants born outside the EU arrived to the 30 nations. Germany, United Kingdom, and Spain received the greatest influx of persons and the majority of migration stemmed from countries in West Asia, the Middle East, and Africa. A significant proportion of total migration was driven by conflict-related crisis in Syria, and East and North Africa. The most significant increases in estimated total hepatitis case numbers, national prevalence increases, and future sequelae were seen in Germany and Sweden. DISCUSSION/SIGNIFICANCE OF IMPACT: Mass migration has significantly changed HBV and HCV disease burden in Europe over the past 5 years. Consequently, long-term outcomes of cirrhosis and HCC are also expected to increase. These increases are likely to disproportionally impact individuals of the migrant and refugee communities. HBV and HCV surveillance and management programs must strategically focus on individuals from high-burden age cohorts and nations. Screening and treatment would aid WHO elimination efforts while benefiting both the vulnerable individuals and host nations through reduction of morbidity, mortality, and associated healthcare expenses.
4112 Tools for Self-Management of Obstetric Fistula in Low and Middle-income Countries: A Qualitative Study Exploring Pre-implementation Barriers and Facilitators among Global Stakeholders
Authors: Ryan Nessa E , Boden-Albala Bernadette , Ganyaglo Gabriel , Ayadi Alison El
Published: July 2020

OBJECTIVES/GOALS: Insertable devices (IDs) for obstetric fistula (OF) management are feasible, acceptable, but not accessible; implementation determinants in low and middle-income countries (LMICs) are unknown. Thus, the purpose of this study was to understand pre-adoption facilitators and barriers among global stakeholders for a therapeutic ID for OF in LMICs. METHODS/STUDY POPULATION: Stakeholders, including researchers (n = 11), clinicians (n = 4), government officials (n = 2), and administrators (n = 4), were purposefully identified from various sectors involved in understanding and addressing the needs of women with OF: clinical care, academia, international health, civil society, and government. Twenty-one individuals were interviewed about their perceptions of IDs for OF self-management and their implementation. Interviews were audio-recorded and transcribed. The Consolidated Framework for Implementation Research (CFIR) guided data collection and analysis. Thematic analyses were carried out within Nvivo v.12. RESULTS/ANTICIPATED RESULTS: Determinants of implementation of an ID for OF self-management (by CFIR domain) include: (1) intervention characteristics—relative advantage and cost; (2) individual characteristics—knowledge and beliefs about the innovation; (3) inner setting-- organizational culture, implementation climate, tension for change, and compatibility; (4) outer setting-- patient needs and resources and external policy and incentives; (5) process—opinion leaders and collaboration. Facilitators include: tension for change for low-cost, accessible IDs; relative advantage over existing tools; development of partnerships; and identification of implementation champions. Barriers include: need for educational strategies to encourage clinical provider acceptability; lack of evidence of the optimal beneficiary. DISCUSSION/SIGNIFICANCE OF IMPACT: Tools for therapeutic OF self-management could be integrated into comprehensive OF programming. Employing the CFIR as an overarching typology allows for comparison across contexts and settings where OF care occurs and may be useful for clinicians, researchers, and policy-makers interested in implementing IDs for OF self-management in LMICs. CONFLICT OF INTEREST DESCRIPTION: I am working with colleagues at the non-profit Restore Health on developing an insertable cup for therapeutic self-management of obstetric fistula in LMICs
4317 Using Failure Modes and Effects Analysis to Guide Adaptation of an Evidence-Based Parenting Program for Mothers with Substance Use Disorders
Authors: Peacock-Chambers Elizabeth , Friedmann Peter , Byatt Nancy , Suchman Nancy , Feinberg Emily
Published: July 2020

OBJECTIVES/GOALS: To identify possible failures that could occur in the delivery of an evidence-based parenting program for mothers with substance use disorders (SUD) through existing home-visiting services, and to develop solutions to the most significant failures. METHODS/STUDY POPULATION: Using failure modes and effects analysis (FMEA) methodology, we conducted two 2-hour advisory panel discussions with 15 people from a variety of disciplines and life experiences related to SUDs. The intervention delivery process included five steps: (1) Recruitment, (2) Screening, (3) Matching, (4) Enrollment in person, and (5) Intervention delivery. Participants collectively determined possible failures, causes, and consequences. Participants then agreed on three scores (Likert Scale 0-10) for the likelihood of occurrence, detection, and severity of the failure, with 10 being the highest likelihood, difficulty detecting, or severity. A risk priority number (RPN) was calculated as the product of the 3 scores (maximum RPN = 1,000). The group then identified possible solutions for failures with higher RPNs. RESULTS/ANTICIPATED RESULTS: For each step in the process we identified the following number of failure nodes and RPN scores: (1) recruitment: 13 failures; RPN = 800, (2) screening: 102 failures; RPN = 10, (3) matching: 4 failures: RPN = 490, (4) enrollment: 6 failures; RPN = 80, (5) delivery: 11 failures; RPN = 80. The most critical failures related to recruitment and were perceived as being caused by potential development of mistrust in the community. Participants strongly encouraged the use of “strengths-based language,” clear referral plans for mothers that did not qualify, and inclusion of mothers that did not have custody of their children. These findings resulted in changes to the screening script, enrollment procedures, and inclusion criterial for the program. DISCUSSION/SIGNIFICANCE OF IMPACT: FMEA methodology was particularly effective in identifying possible failures for the integration of an evidence-based parenting program into existing home-visiting services as they related to the psychological safety of mothers with SUDs. The process resulted in direct changes to procedures for the anticipated program integration and study.
4053 A TL1 Team Approach to CNS-Localized Delivery of Glial Cell-Derived Neurotrophic Factor for Treatment of Parkinson’s Disease
Authors: Farhadi Shaheen , Gopinath Adithya , Streit Wolfgang , Hudalla Gregory A , Khoshbouei Habibeh
Published: July 2020

OBJECTIVES/GOALS: Develop a strategy to restrict GDNF diffusion at an injected CNS tissue site for dopamine neuron rescue by endowing it with binding affinity for carbohydrates that are abundant on the cell surface and extracellular matrix. METHODS/STUDY POPULATION: GDNF will be fused to galectin-3 (G3), a human protein that binds to β-galactoside residues of cell surface and matrix glycoproteins. We characterized the binding of G3 fusion proteins to various glycoproteins and primary human myeloid cells. We incubated G3 fusions with CNS tissue ex vivo to measure their binding and depth of penetration via diffusion. We next plan to administer GDNF-G3 via CNS intracranial infusion in a murine PD model and then conduct behavioral PD phenotype testing via rotarod and pole descent to compare to non-parkinsonian controls. We will further examine the effects of GDNF-G3 on degeneration using immunohistochemical examination of post-mortem brain tissue. RESULTS/ANTICIPATED RESULTS: Based on results from previous clinical trials of GDNF delivery, we anticipate that a successful intervention using GDNF-G3 will result in rescue of midbrain dopaminergic neurons in a murine PD model. In murine CNS tissue, we observed binding to glycans at the tissue surfaces when incubated with G3 fusion proteins ex vivo, suggesting GDNF-G3 will remain localized to the injection site. Next we will administer GDNF-G3 via CNS intracranial infusion in a murine PD model and assess efficacy by behavior and histopathology. GDNF-G3-mediated dopamine neuron rescue are expected to slow or reverse the progression of PD in these animal models. DISCUSSION/SIGNIFICANCE OF IMPACT: PD treatments focus on symptomatic relief. Standard therapies have not been efficacious in rescuing of dopaminergic neurons. GDNF-G3 administered at the site of neurodegeneration would represent a milestone on the path to treating PD pathology and address limitations of GDNF delivery.
4430 Coffee Shops and Fast-food Restaurants: Potential Neighborhood Resources for Cognitive Health and Wellbeing Among Aging Americans
Authors: Finlay Jessica , Esposito Michael , Tang Sandra , Gomez-Lopez Iris , Sylvers Dominique , Judd Suzanne , Clarke Philippa
Published: July 2020

OBJECTIVES/GOALS: Environmental factors may significantly increase the risk of or buffer against Alzheimer’s disease and related dementias, yet strategies to address cognitive decline and impairment to date largely overlook the role of neighborhoods. This mixed-methods study is the first to examine potential links between access to eateries and cognitive function. The goal is to inform place-specific interventions to help aging individuals reduce risk for cognitive impairment through neighborhood community and design. METHODS/STUDY POPULATION: Following an exploratory sequential mixed-methods design, seated and mobile interviews with 125 adults aged 55-92 (mean age 71) living in the Minneapolis (Minnesota) metropolitan area suggest that eateries, including coffee shops and fast-food restaurants, represent popular neighborhood destinations for older adults and sources of wellbeing. To test the hypothesis that these sites, and the benefits they confer, are associated with cognitive welfare, we analyzed data from urban and suburban dwelling participants in the REasons for Geographic And Racial Differences in Stroke (REGARDS) study, a national racially diverse sample of older Americans followed since 2003 (n = 16,404, average age at assessment 72 years). RESULTS/ANTICIPATED RESULTS: Qualitative thematic analysis of how older adults perceived and utilized local eateries include sites of familiarity and comfort; physical and economic accessibility; sociability with friends, family, staff, and customers; and entertainment (e.g., destinations for outings and walks, free newspapers to read). Quantitative results from multilevel linear regression models demonstrate a positive association between density of eateries and cognitive functioning. Taken together, these results complicate our understanding of fast-food settings as possible sites of wellbeing through social interaction and leisure activities. DISCUSSION/SIGNIFICANCE OF IMPACT: The results contribute new evidence towards an emerging ecological model of cognitive health. Understanding whether and how retail food environments can help buffer against cognitive decline among older adults provides novel opportunities to promote wellbeing in later life through community interventions and neighborhood design.
4414 Collagen Dermal Replacement Scaffold Mechanobiology in Treatment of Difficult-to-Heal Wounds
Authors: Sohutskay David Oleh , Tepole Adrian Buganza , Voytik-Harbin Sherry
Published: July 2020

OBJECTIVES/GOALS: Difficult-to-heal wounds of the skin are a common and costly medical problem. Dermal replacement strategies have emerged as a solution, but a challenge is identification of optimal scaffold parameters. We present a model for assessment of clinical potential of collagen scaffolds for wound healing. METHODS/STUDY POPULATION: In previous animal experiments, we evaluated dermal replacement scaffolds custom-fabricated from fibril-forming collagen oligomer with controlled fibril density (4, 20, 40mg/cm3) and spatial gradients in rat excisional wounds. Wound contraction and cellularization were monitored by gross and histological image analysis for comparison with model outcomes. We now parameterize the scaffold parameters for use in the mathematical model of wound healing with nonlinear curve fitting. A preliminary chemo-bio-mechanical finite element model including collagen, cells, and an inflammatory signal was adapted to simulate wound healing results. RESULTS/ANTICIPATED RESULTS: Collagen oligomer microstructure was quantified from scanning electron micrographs. A constitutive law for collagen mechanics was fit to experimental uniaxial tensile tests. We have conducted preliminary three-dimensional finite element model simulations to be validated against experimental wound contraction, recellularization, and collagen remodeling data collected from each experimental group. We show the effects of collagen density and stiffness on wound contraction by altering early wound mechanical properties. We anticipate future work to further improve the model of mechanotransduction, inflammation, and recellularization. DISCUSSION/SIGNIFICANCE OF IMPACT: This work represents the first step towards a computational model of wounds treated with collagen scaffold dermal replacements. In turn, the model will be used to explore cell-scaffold interactions for purposes of prediction and optimization of tissue regeneration outcomes.
4026 Dissecting the role of microenvironment heterogeneity on metastatic tumor cell phenotype at an engineered metastatic niche
Authors: Orbach Sophia , Brooks Michael D. , Bushnell Grace G. , Wicha Max S. , Jeruss Jacqueline S. , Shea Lonnie D.
Published: July 2020

OBJECTIVES/GOALS: Breast cancer metastases are stochastic and difficult to detect. Therapy is often ineffective due to phenotypic changes of tumor cells at these sites. We engineered a synthetic metastatic niche to study the role of phenotypic transitions in the microenvironment on tumor cell phenotype. METHODS/STUDY POPULATION: The engineered metastatic niche is composed of a porous polycaprolactone scaffold implanted subcutaneously in Balb/c mice. The mice received an orthotopic inoculation of 4T1 cells (murine triple negative breast cancer) in the fourth right mammary fat pad and the disease was allowed to progress for 7-21 days (pre-metastatic to overt metastatic disease). The scaffolds and lungs (native metastatic site) were explanted and analyzed by single cell RNA-seq via Drop-seq. Cell phenotypes were identified and tracked over time with the Seurat and Monocle3 pipelines. Assessment of the impact of these cell populations on tumor cell phenotype was conducted through Transwell co-cultures. RESULTS/ANTICIPATED RESULTS: Healthy scaffolds are primarily composed of macrophages, dendritic cells, and fibroblasts – consistent with a foreign body response. Despite differences in the lung and scaffold prior to tumor inoculation, both tissues were marked by >5-fold increase in neutrophils/MDSCs. Additionally, 79% of genes at the scaffold that significantly changed over time were also identified in the lung, indicating key similarities in niche maturation. However, many immune cells at the scaffold had distinct phenotypes, with pro-inflammatory/cytotoxic characteristics. These changes clearly impacted tumor cell phenotype, as cells from the scaffold increased tumor cell migration and apoptosis in vitro. DISCUSSION/SIGNIFICANCE OF IMPACT: Early phenotypic changes at the engineered metastatic niche can identify signs of metastasis prior to colonization of tumor cells. Furthermore, dynamics of immune and stromal cells change throughout niche maturation, influencing tumor cell phenotype and may suggest targeted therapies. CONFLICT OF INTEREST DESCRIPTION: Lonnie Shea, Jacqueline Jeruss, and Grace Bushnell are named inventors on patents or patent applications.
4241 Identification of small molecules that facilitate the efficient differentiation of stem cell derived β-cells
Authors: Min Yuhao , Clifford Chris , Peterson Quinn P.
Published: July 2020

OBJECTIVES/GOALS: In this study, we established a high-throughput chemical screening platform to identify small molecules that facilitates efficient differentiation of stem cells derived β (SC-β) cells. Using this platform, we identified several compounds that potentially increase the differentiation efficiency. METHODS/STUDY POPULATION: Differentiation of human embryonic stem cells (HUES8) into SC-β was carried out using previously published protocols in a 3D cell suspension. Single cells were replated in Matrigel-coated well plates at the start of different stages depending on experiments. Differentiation medium supplemented with small molecules at a final concentration of 2 M and 0.2 M was used throughout the stage. All the cells were then fixed and permeabilized. Immunocytochemical staining was performed. Images of each well were taken and analyzed. Numbers of the total cell, insulin-positive cell, NKX6.1-positive cell, and co-positive cell were recorded. Candidate compounds were validated using flow cytometry or ICC. RESULTS/ANTICIPATED RESULTS: We identified several hit compounds that significantly increase the NKX6.1 positive cell percentage compared to the DMSO-treated controls when treated at the PP1 cell stage. Follow up assays demonstrated that at least one of these putative hits reproducibly increased NKX6.1 expression. In addition, we identified other compounds that significantly increase the insulin and NKX6.1 copositive SC-β cell population when treated at the later PP2 cell stage during the differentiation. We expect a dosage-dependent response when the candidate hits are validated using more accurate assays. DISCUSSION/SIGNIFICANCE OF IMPACT: We established a high-throughput screening platform to identify small molecules that increase the efficiency of SC-β direct differentiation. Successful generation of SC-β allows cell replacement therapy in diabetes patients, and a better understanding of pancreatic biology and development.
4361 Interim Results of the Effect of Haptoglobin Phenotype on Inflammatory Cytokine Concentrations in Plasma and CSF After Aneurysmal Subarachnoid Hemorrhage
Authors: Smith Austin
Published: July 2020

OBJECTIVES/GOALS: Haptoglobin (Hp) phenotypes may affect inflammatory response after neurologic injury. We are investigating the relationship between patient Hp phenotypes and inflammatory cytokine concentrations in plasma and CSF after aneurysmal subarachnoid hemorrhage (aSAH), a severe form of hemorrhagic stroke. METHODS/STUDY POPULATION: Following IRB approval, all patients with angiographically-proven aSAH and who underwent extraventricular drain (EVD) placement were included. Patients were excluded if they were not expected to survive hospitalization or were on pre-existing anti-inflammatory agents. For all enrolled patients, plasma and CSF samples were taken on post-bleed days 3, 5, 7, and 10, processed and then frozen for later analysis. In this interim analysis, Hp phenotype was assessed through ELISA analysis of plasma samples and cytokine concentrations were determined using multiplex ELISA kits for both plasma and CSF samples. Hp phenotypes were dichotomized to either HP1 (Hp 1-1 or 1-2) or HP2 (Hp 2-2). RESULTS/ANTICIPATED RESULTS: To date, 23 aSAH patients have been enrolled in this IRB-approved study. An interim analysis of the first 13 patients has revealed eight Hp1 patients (Hp 1-1 n = 1, Hp 1-2 n = 7) and five Hp2 patients. CSF levels of IL-6 and TNF-a were greater than plasma levels in all patients at all time points. CSF levels of IL-6 appear to peak on PBD 5 (1890 ± 767 pcg/mL) and 7 (1612 ± 899 pcg/mL). The CSF IL-6 concentrations in the Hp2 group were lower (1792 ± 806 pcg/mL vs. 1952 ± 791 pcg/mL) on PBD5 but were higher (1635 ± 930 pcg/mL vs 1598 ± 943 pcg/mL) at PBD7; however, these differences did not reach statistical significance. DISCUSSION/SIGNIFICANCE OF IMPACT: This interim analysis demonstrated no statistically significant differences in plasma or CSF cytokine concentrations between patients with different Hp phenotypes. This may be due to the low number of samples or the potential confounding effect of disease-specific secondary neurological injuries.
4222 Synthesis and application of cyanuric chloride lipids for peptide presentation
Authors: Nardo David , Venditto Vincent J.
Published: July 2020

OBJECTIVES/GOALS: My long-term career objective is to become an established independent researcher focused on understanding and modulating immune responses to biologics in order to enhance their efficacy and understand the underlying mechanisms by which these interact with the immune system. METHODS/STUDY POPULATION: In this study we will evaluate the utility of cyanuric chloride based synthetic lipids in the presentation of peptide epitopes of the gene delivery vector, adeno-associated virus (AAV). The lipopeptide conjugates will be administered to mice via liposomal formulations to assess their ability to induce immune responses by ELISA as compared to mice treated with the AAV. The three-dimensional conformation of the peptides will be evaluated using nuclear magnetic resonance to determine their similarity with the natural conformation that these peptides adapt on the viral surface. Additionally, to assess the translatability of this conjugation strategy, we will test the ability of our lipopeptides to bind to serum antibodies from human subjects. RESULTS/ANTICIPATED RESULTS: We anticipate that peptide presentation using cyanuric chloride lipids will achieve a robust response in mice following immunization. Immunizations with our lipids should induce the production of antibodies targeting AAV, albeit a milder response that the virus itself, given the complexity of viral epitopes. Nuclear magnetic resonance will inform us on how to improve the synthetic lipids to optimize peptide presentation by altering the characteristics of the lipid anchors. Finally, by using human serum to test for the ability of our lipopeptides to bind to antibodies in serum from patients positive for AAV antibodies, we can become informed on whether our strategy has utility in human studies or whether our method is limited to animal models of human disease. DISCUSSION/SIGNIFICANCE OF IMPACT: The current work seeks to develop a strategy to present peptides from a well characterized biologic, AAV, on a liposome surface. Our ultimate purpose is to employ liposomal formulations as decoys that target AAV-specific lymphocytes to improve the in vivo efficacy of AAV.
Androgen receptor in satellite cells is not essential for muscle regenerations
Authors: Sakai Hiroshi , Sato Takahiko , Kanagawa Motoi , Fukada So-ichiro , Imai Yuuki
Published: July 2020

The anabolic effects of androgen on skeletal muscles are thought to be mediated by androgen receptor (AR). Although multiple studies concerning the effects of AR in males have been performed, the molecular mechanisms of AR in skeletal muscles remain unclear. Here we first confirmed that satellite cells from mouse hindlimb muscles express AR. We then generated satellite cell-specific AR knockout mice using Pax7CreERT2 and ARL2/Y mice to test whether AR in satellite cells is necessary for muscle regeneration. Surprisingly, we found that muscle regeneration was compromised in both Pax7CreERT2(Fan)/+ control mice and Pax7CreERT2(Fan)/+;ARL2/Y mice compared to ARL2/Y mice. However, Pax7CreERT2(Gaka)/+;ARL2/Y;R26tdTomato/+ mice showed no significant differences between control and mutant muscle regeneration. These findings indicate that AR in satellite cells is not essential for muscle regeneration. We propose that Pax7CreERT2(Fan)/+ control mice should be included in all experiments, because these mice negatively affect the muscle regeneration and show the mild regeneration phenotype.
Welfare Quality®for dairy cows: towards a sensor-based assessment
Authors: Maroto Molina Francisco , Pérez Marín Carlos C. , Molina Moreno Laura , Agüera Buendía Estrella I. , Pérez Marín Dolores C.
Published: July 2020

This Research Reflection addresses the possibilities for Welfare Quality®to evolve from an assessment method based on data gathered on punctual visits to the farm to an assessment method based on sensor data. This approach could provide continuous and objective data, while being less costly and time consuming. Precision Livestock Farming (PLF) technologies enabling the monitorisation of Welfare Quality®measures are reviewed and discussed. For those measures that cannot be assessed by current technologies, some options to be developed are proposed. Picturing future dairy farms, the need for multipurpose and non-invasive PLF technologies is stated, in order to avoid an excessive artificialisation of the production system. Social concerns regarding digitalisation are also discussed.
The Development of PRIMA - A Belgian Prediction Model for Patient Encounters at Mass Gatherings
Authors: Spaepen Kris , Haenen Winne AP , Hubloue Ives
Published: July 2020

Introduction:

Mass gatherings (MGs) grow in frequency around the world. With the intrinsic potential for significant health risks for all involved, MGs pose a challenge for those responsible for the provision of on-site medical care. Belgian law obliges local governments to identify and analyze the risks involving a MG. Though medical risk factors are long known, all too often, resourcing for in-event health services is based on anecdotal and previous experiences.

Problem:

Despite the fast-evolving science on MGs, the lack of reliable tools – based on empirical and analytical approaches – to predict patient presentation rates (PPRs) at MGs remains.

Methods:

A two-step method was followed to develop, update, and support a Plan Risk Manifestation (PRIMA) program. First, a continuous systematic literature review was conducted. Once developed, the model was run using data obtained from Belgian Federal Public Service (FPS; Brussels, Belgium) Health, Food Chain Safety, and Environment (HFCSE); event organizers; and municipalities.

Results:

In total, 231 studies and documents were included to form the program. With the data provided, three variables were computed to run the calculation model to predict the PPR. Three medical risk axes were defined for this model: (1) isolation risk; (2) population risk; and (3) risk at illness. A combined dataset was derived from the prediction of the PRIMA program combined with the actual data obtained after the MG. This proved a solid basis for the calculation model of the PRIMA program.

Conclusion:

Despite that validation is needed, the PRIMA program and its prediction model for PPRs at MGs carries the promise of a general, applicable prediction and risk analysis tool for a multitude of events.
Polarization-controlled optical holography using flat optics
Authors: Majumdar Arka , Colburn Shane
Published: July 2020

Due to the large number of degrees of freedom offered by nanoscale scatterers, a single flat optic can project different images at different distances depending on the polarization of the light, opening up opportunities for optical encryption and augmented reality systems.