At NERIC2023, we are excited to feature abstracts from IDeA students, target and pilot investigators, core facilities and those studying data science regardless of funding source.  Below are some of the abstracts which will be presented as posters at our poster sessions.  Please note that not all abstracts are available here on the website due to author requested embargo.  

*Indicates Presenting Author

Predicting Extension in Juvenile Idiopathic Arthritis
Megan Simonds* (Nemours Children’s Health), Kathleen Sullivan (The Children’s Hospital of Philadelphia), AnneMarie Brescia (Nemours Children’s Health)

Background: Juvenile idiopathic arthritis (JIA) is the most common rheumatic disease of childhood and carries a risk of permanent joint damage and disability when patients evolve from a persistent (P) course to a polyarticular course, termed extended (E) [1]. Our objective is to develop a robust panel of biomarkers to predict which patients will extend.
Methods: We performed ELISA on 7 biologically relevant markers: CD14, angiogenin, IL-5, VEGF, IL-1α, IL-6 and IL-3 on synovial fluid samples from our Nemours IRB-approved repository and Cincinnati Children’s Hospital Medical Center (CCHMC). Nemours BioBank collected blood serum samples from patients with JIA and we measured our biomarkers using Luminex in our synovial fluid and serum samples. scRNA-seq was performed by Genewiz using 10x Genomics Chromium protocols on fibroblast-like synoviocytes. 
Results: From our Nemours cohort, there was reasonable specificity for all the biomarkers with CD14 being the most specific marker (86%); however, IL6 (71%) was the most sensitive for predicting which synovial fluid came from patients who ultimately extended (Figure 1A). When we examined the CCHMC cohort, CD14 remained the most specific (65%) and IL6 the most sensitive (56%) (Figure 1B). This remained true when we combined the data from the two cohorts (Figure 1C). Next, we developed Luminex assays, a method for detecting multiple proteins in a single sample in a high-throughput manner and successfully detected our markers in both synovial fluid and serum using this method. Lastly, scRNA-sequencing data revealed differentially expressed genes that distinguish between subtypes of JIA [2]. 
Conclusions: Based on our results, these biomarkers are better for predicting who will remain persistent as opposed to predicting those who will extend. Proteins encoded by genes from our scRNA data can be used to expand our biomarker panel. Detecting these proteins in serum using Luminex allows for a more clinically applicable approach. 
1. Manners, P.J. and C. Bower, Worldwide prevalence of juvenile arthritis why does it vary so much? The Journal of rheumatology, 2002. 29(7): p. 1520-1530.
2. Simonds M, S.K., Rose C, Brescia A. . Heterogeneity of Juvenile Idiopathic Arthritis Synovial Fibroblasts Correlates to Disease Progression and Provides Compelling Diagnostic Data [abstract]. Arthritis Rheumatol 2022  [cited 73 May 19, 2022:[Available from: https://acrabstracts.org/abstract/heterogeneity-of-juvenile-idiopathic-arthritis-synovial-fibroblasts-correlates-to-disease-progression-and-provides-compelling-diagnostic-data/.
Chronic Caffeine Consumption Curbs rTMS-Induced Plasticity
Megan Vigne* (Brown University), Jamie Kweon(Brown University, Harvard University), Prayushi Sharma(Brown University, Harvard University), Benjamin D. Greenberg(Brown University), Linda L. Carpenter(Brown University), Joshua C. Brown(Brown University, Harvard University)

Background: Caffeine is a widely used psychostimulant. In the brain, caffeine acts as a competitive, non-selective adenosine receptor antagonist of A1 and A2A, which are known to modulate long-term potentiation (LTP), the cellular basis of learning and memory. Repetitive transcranial magnetic stimulation (rTMS) is theorized to work through LTP induction, and can modulate cortical excitability as measured via motor evoked potentials (MEP).
Methods: To evaluate the effect of chronic caffeine on brain plasticity, we conducted a post-hoc covariate analysis from two pharmaco-rTMS studies in healthy subjects using NMDA partial agonist, D-cycloserine (DCS). We sampled a total of 20 healthy individuals from two sites (ages 21-39, 10 female), 16 of them being regular caffeine users. Single pulse (SP) MEP data was collected over a 1-hour time course following a plasticity-inducing rTMS session.
Results: In this hypothesis-generating pilot study, we observed enhanced MEP facilitation in non-caffeine users compared to caffeine users and to both groups in the placebo condition.
Conclusion: These preliminary data highlight a need to directly test the effects of caffeine in prospective well-powered studies. In theory, they suggest that chronic caffeine use could limit plasticity, perhaps even including rTMS effectiveness.
Cell-type Dependent Differences in JC Polyomavirus Signaling Pathway Regulation
Melissa Maginnis* (Univ of Maine), Michael Wilczek (Univ of Maine), Francesca Armstrong (Univ of Maine), Colleen Mayberry (Univ of Maine), and Benjamin King (Univ of Maine)

JC polyomavirus (JCPyV) infects the majority of the population and causes an incurable persistent infection in the kidneys. In immunocompromised individuals, JCPyV can become reactivated in the central nervous system and infect glial cells, oligodendrocytes and astrocytes, which are critical for myelin production. The molecular mechanisms of JCPyV infection of astrocytes are poorly understood, in part due to most studies being limited to an immortalized cell model. To better understand the cellular and molecular basis of JCPyV infection in astrocytes, we developed and characterized a new infection model using normal human astrocytes (NHAs). We determined that viral infection was regulated differently in primary and immortalized cell types, due to viral factors, such as T antigen expression, and cellular factors, including activation of cellular signaling pathways. Using RNA Sequencing analysis and complementary molecular approaches, we defined cell-type specific differences in the regulation of the MAPK pathway through dual-specificity phosphatases (DUSPs) and also determined the importance of the PI3K/AKT signaling pathway in NHAs. Through this comparative genomic analysis, we elucidated how JCPyV orchestrates differential gene expression and regulation of cellular signaling pathways to mediate infection in primary astrocytes. Outcomes of this research provide an enhanced understanding of cell-type dependent differences in viral infection and can be applied to our broader understanding of cell signaling pathways and future development of antiviral therapies.
Small molecule intervention against Drosophila models of sporadic and familial Parkinson’s disease
Dionne Williams (Delaware State University), Angeline-Claudia Atheby (Delaware State University), Hakeem Lawal* (Delaware State University)

Parkinson’s disease (PD) is the second most common neurodegenerative disease. Decades of research has established key environmental and genetic factors as contributors to its etiology although the precise cause of most PD cases remains unknown. Moreover, despite the advances in our understanding of the possible causes of PD, a viable treatment remains elusive. Both rotenone, a potent laboratory model for sporadic PD, and alpha synuclein over or misexpression have been used to uncover important insights into the etiology of the disease. We are interested in developing potential new disease-modifying therapeutic strategies against PD. To this end, we are testing the neuroprotective capability of the small molecule dacarbazine, a chemotherapeutic that has been identified as a potential anti-PD agent. We investigated whether the compound is capable of conferring protection against rotenone toxicity. We report that dacarbazine protects against organismal and neuronal toxicity induced by rotenone exposure in both male and female Drosophila. We also report preliminary studies on the effect of this compound in alpha synuclein model of both neuronal and organismal toxicity. Together, these findings provide key evidence of a neuroprotective capability of a putative anti-PD agent against models of PD.
Engaging Community Partners to Co-Develop a Multi-Component Intervention to Increase Equitable Uptake of COVID-19 Vaccines
Paul Enlow* (Nemours Children’s Hospital, Delaware), Courtney Thomas (Nemours Children’s Hospital, Delaware), Marshala Lee (Christiana Care Health System), Jonathan Miller (Nemours Children’s Hospital, Delaware), Lavisha Pelaez (Nemours Children’s Hospital, Delaware), Anne Kazak (Nemours Children’s Hospital, Delaware), Thao-Ly Phan (Nemours Children’s Hospital, Delaware)

Introduction: The COVID-19 pandemic exacerbated health disparities across the United States. Although non-Hispanic Black (Black) and Latinx youth experienced higher rates of COVID-19 transmission, morbidity, and mortality, these same youth are less likely to receive the COVID-19 vaccine. In this presentation we describe the process of engaging community partners in co-developing an intervention to promote equitable uptake of the COVID-19 vaccine.
Method: Our process for engaging community partners was informed by the Promising Practices Guide developed by New Mexico’s Engage for Equity Team. With help from advisors with expertise in community engagement, we assembled a team of 11 stakeholders from Black or Latino communities. We co-developed intervention strategies over the course of eight meetings. We then used mixed-methods crowdsourcing approach with Black and Latinx youth (n=15; Mage=13.9+1.6 years) and caregivers (n=20; Mage=40.4+7.3 years) to elicit perspectives on the acceptability of these intervention strategies. Youth and caregivers viewed intervention strategies, indicated if they found the strategy acceptable, and explained their rating in open-ended responses. Strategies that <80% of participants rated as acceptable were revised based on open-ended responses.
Results: We co-developed five intervention strategies: (1) community-tailored handouts/flyers, (2) informational videos featuring local families, (3) family-centered language to offer vaccines, (4) communication-skills training for providers, and (5) community health workers. All strategies were rated as acceptable by ≥80% of participants, except for “family-centered language to offer vaccines.” Open-ended responses revealed that families disliked some of the wording and wanted to ultimately decide if their child received the COVID-19 vaccine.
Conclusions: Engaging community stakeholders yielded acceptable strategies to promote equitable uptake of the COVID-19 vaccine. Utilizing the empirically-supported methods found in the Promising Practices Guide can enhance the quality of stakeholder engagement. We will discuss lessons learned from engaging community partners in this project and how to apply this process to other populations.
Introducing the Intervention Methodology: Provision and Connection through Technology (IMPACT) Core: A Research Core supported by the Research Expanding Access to Child Health (REACH) Center
Michelle Goodreau* (Nemours), Kimberly Canter (Nemours)

The REACH Center is an NIH Center for Biomedical Research Excellence (COBRE P20GM144270) at Nemours Children’s Hospital, Delaware. The IMPACT Core is supported by the REACH Center and is a new resource for investigators who are interested in conducting innovative intervention research focused on reducing pediatric health disparities and improving child health. IMPACT has an emphasis on technology-enhanced intervention research and research utilizing novel methodological approaches. IMPACT is driven by several aims to further its development.  The first aim of the IMPACT Core is to create a robust infrastructure to facilitate the development, delivery and implementation of scalable, innovative, technology-enhanced interventions primed for delivery to patients and families impacted by health disparities and social inequities. A Core Internal Advisory Committee and a group of experts in various types of technology support the Core. Additionally, an intake form was developed in REDCap and disseminated to foster relationships in Delaware and log results from ongoing consultation meetings.
The second aim of IMPACT is to prepare and support investigators in the conduct of intervention research using technology to address equity in pediatric health care. In addition to supporting research and pilot projects funded by the REACH Center, the Core Director and other Core personnel consult with investigators across the state. These consultations have been used to assist with the development of pilot and research projects, manuscript preparations, and an R21 application.  Our final aim is to manage and sustain the IMPACT Core. This includes investments in training opportunities and incorporating cutting edge technological and methodological developments into interventions supported by REACH. To ensure continued success, we are tracking our progress with multiple projects and consultations and continuing to have regular meetings with our collaborators. IMPACT is facilitating a workshop in September 2023, in partnership with The Center for Health Delivery Innovation at Nemours.
Imaging of Neurofibromatosis Type 1 with Two Fluorine-18 Labeled Radiotracers
Shriya Boyapati (Nemours Children’s Health), Sigrid A. Langhans (Nemours Children’s Health), Thomas H. Shaffer (Nemours Children’s Health), Wenqi Xu (Nemours Children’s Health), Vinay V.R. Kandula (Nemours Children’s Health), Heidi H. Kecskemethy (Nemours Children’s Health), Lauren W. Averill (Nemours Children’s Health), Rahul M. Nikam (Nemours Children’s Health), Xuyi Yue* (Nemours Children’s Health)

Introduction: Neurofibromatosis type 1 (NF1) is a rare autosomal disorder. Eight to 16% of children with a benign form of NF1 develop malignant peripheral nerve sheath tumors (MPNSTs) with a dismal overall 5-year survival rate of approximately 30%. Dysregulated tryptophan metabolism has been associated with malignant progression and overall survival of human NF1. We aim to use two fluorine-18 labeled radiotracers, 1‐(2‐[18F]fluoroethyl)‐L‐tryptophan (L-[18F]FETrp) and F-18 Fluorodeoxyglucose (FDG), to perform dynamic scans in a transgenic NF1 animal model and compare the imaging results.
Methods: L-[18F]FETrp was radiosynthesized in-house. The mice subjected to one-hour dynamic PET scans were divided into four groups (n = 4/group). 3D regions of interest (ROIs) were drawn manually. Standardized uptake value (SUV) was plotted. The target to non-target ratios were calculated and compared. H&E staining was performed in wild-type and lesioned samples.
Results: L-[18F]FETrp was synthesized in 100 min with good yields. In the wild-type group, mice showed high uptake in the heart and the bladder for both tracers. Tumors typically develop at 3 – 5 months. Representative mean SUV showed the tumor had comparable uptake between L‐[18F]FETrp and FDG. The brain region had substantially less uptake for L‐[18F]FETrp. The PET scans with L‐[18F]FETrp and FDG showed significant differences in tumor-to-brain ratios (4.29 ±1.38 vs. 1.03 ± 0.75 at 5 min, 2.66 ± 0.95 vs. 1.00 ± 0.57 at 45 min, n = 4, p < 0.05). H&E staining showed morphological differences between the healthy and lesioned samples.
Conclusion: L‐[18F]FETrp and FDG showed statistically different kinetics in the heart of wild-type mice. L‐[18F]FETrp had significantly more favorable tumor-to-brain SUV ratios than FDG. The application of L‐[18F]FETrp in neuro-oncology is promising due to low tracer uptake in non-target mice brains.
Impact of SARS-COV-2 Infection in Children on Cardiac Involvement – a temporal trend assessment: preliminary pilot data from the N3C Data Enclave
Shubhika Srivastava* (Nemours), Julie Cowart (University of Delaware), Suzanne McCahan (Nemours), Jobayer Hossain (Nemours), Ran Zhang (Nemours), Sai Prashanthi Gumpili (Prashanthi Gumpili), Carol Prospero (Nemours), Cathy Wu (University of Delaware), Robert Akins (Nemours), Claudine Jurkovitz (Christiana Care Health System), on behalf of the N3C Consortium

Background: Data on prevalence and trends of cardiac involvement in children and outcomes following SARS- COV-2 infection are lacking. The aim of this study is to examine the temporal occurrence of cardiac outcomes within 6 months after the initial COVID-19 diagnosis.
Method: We identified patients age 0-20 years who had positive lab test or diagnosis for COVID-19 prior to October 1, 2021 within the National COVID Cohort Collaborative (N3C) Data Enclave, a repository of 73 sites across the U.S. with over 5,000,000 adult and pediatric COVID positive cases. Quarterly temporal trends of cardiac conditions were calculated during the 6 months after initial COVID-19 infection. Per N3C policy, groups with <20 patients cannot be reported. Instead, we calculated the percentages representing 20 patients and reported that the value is less than that percent. Congestive heart failure, cardiomyopathy, myocarditis, pericarditis, pericardial effusion, arrhythmias, and Kawasaki disease were identified using SNOMED/ICD codes concept sets defined in the N3C enclave. Patients with pre-existing cardiac conditions were not excluded from the analysis.
Results: The study population included 595,738 patients. Of those, 51% were female, 16% were age 0-4 years at the time of infection, 24% were 5-11, 33% were 12-17, 26% were 18-20, 53% were white, 13% were black, 18% were Hispanic or Latino. Of the patients, 2.2% were hospitalized within 1 day prior and 16 days after initial COVID-19 infection. Temporal trends of cardiac diagnosis are noted in Table 1 – ranging from less than 0.01% to 0.49% with a peak – in cardiac diagnoses in Q2 of 2020 with arrhythmia, congestive heart failure and cardiomyopathy being the most common overall.
Conclusion: These are preliminary pilot data that will allow assessment of population burden and temporal trends in cardiac diagnoses in children. This study is being designed to develop a risk prediction model to ascertain cardiac involvement and resource needs.
Association between Mediterranean diet and urine metal mixtures concentrations in pregnant women from the New Hampshire Birth Cohort Study.
Leyre Notario Barandiaran* (Dartmouth College), Jose A. Signes Pastor (Universidad Miguel Hernandez de Elche), Margaret R. Karagas (Dartmouth College)

During pregnancy, an adequate exposure to essential metals with reduced exposure to non-essential ones is key for proper fetal growth and development and maternal health. Diet is among the major source of metals exposure. However, little is known about the impact of exposure to real-life metal mixtures from dietary patterns such as Mediterranean diet, which is known for its established health benefits. The aim of this study was to cross-sectionally investigate the association between adherence to the Mediterranean diet and metals exposure individually and as a mixture. This study used a sample of 1,068 pregnant women from the New Hampshire Birth Cohort Study (NHBCS). We used a validated food frequency questionnaire to calculate one of the most widely used Mediterranean diet indexes, the relative Mediterranean Diet (rMED). This index is composed of 8 traditional Mediterranean components with a range of scores from 0-16 points. Additionally, we measured aluminum (Al), cadmium (Cd), cobalt (Co), copper (Cu), iron (Fe), mercury (Hg), molybdenum (Mo), nickel (Ni), lead (Pb), antimony (Sb), selenium (Se), tin (Sn), zinc (Zn), and arsenic (As) speciation in urine samples collected at 24-28 weeks of gestation as exposure biomarkers using inductively coupled plasma mass spectrometry. We performed multiple linear regression and bayesian kernel machine regression to analyze the association between rMED and exposure to the metal mixture. High adherence (11-16 points) to Mediterranean diet was associated with increased urinary Al and AsB with  = 0.34 (95%CI = 0.06; 0.61) and  = 1.24 (95%CI = 0.72; 1.76), respectively, compared to low adherence (0-6 points). Intake of fruits and nuts, fish, and olive oil were the rMED components responsible for the results obtained with the total rMED score. The Mediterranean diet is a healthy dietary pattern for pregnant women in the US. However, efforts should be focused on promoting and educating the population in the choice of healthy products within the Mediterranean diet, prioritizing fresh over canned products, as well as choosing quality vegetable oils, to avoid unnecessary exposure to metals.
Neonatal pain and its impacts on the cellular physiology and neural heterogeneity of the central nucleus of the amygdala (CeA)
Megan Tomasch* (University of Maine/University of New England), Michael Burman (University of Maine/University of New England)

Painful and traumatic experiences during development, such as time spent in the neonatal intensive care unit (NICU), have been shown to increase susceptibility to pain- and anxiety-disorders later in life. Using a rodent model of a typical NICU experience, our lab previously associated this pain-vulnerability with changes to cells expressing corticotropin releasing factor (CRF) in the central nucleus of the amygdala (CeA). The CeA is a hub of emotional responding comprised of a functionally heterogeneous population of GABAergic neurons defined by expression of biomarkers such as CRF, somatostatin (SOM), dynorphin (DYN), protein kinase C delta (PKC-δ). These cellular populations play distinct roles in anxiety, fear, and pain, but current research often fails to consider co-expression of multiple markers, including with CRF, and the impacts of neonatal trauma. We hypothesize that neonatal trauma alters the composition and function of biomarker-identifiable subpopulations within the CeA-CRF system, creating a pain-induced neural plasticity that primes the subjects for altered pain responses and anxiety-like behaviors in later-life. Using fluorescent in situ hybridization (fISH), we visualized expression of CRF, SOM, and DYN, on post-natal days (PD) 12, 24, and 48 in male and female rats that either experienced neonatal pain or were left undisturbed. Preliminary data suggest age-dependent changes in the number, biomarker-phenotype, and patterns of biomarker co-expression. To further understand these changes, we performed acute-slice patch-clamp electrophysiology in transgenic rats expressing TdTomato in CRF+ neurons. We find that CeA-CRF+ cells display at least three distinct firing patterns (e.g., regular spiking, late firing, and burst firing) that exhibit different cellular characteristics. Furthermore, data suggest that neonatal trauma may lead to an altered rheobase and current threshold. Future analysis using post-hoc immunohistochemistry of patch-clamped cells will link these characteristics with specific biomarkers and examine how trauma/pain and development impact these biomarker-identified subpopulations.
Effects of Maternal Triclosan Exposure on the Infant Microbiome
Aislinn Gilmour* (Geisel School of Medicine at Dartmouth College), Megan Romano (Geisel School of Medicine at Dartmouth College), Juliette Madan (Geisel School of Medicine at Dartmouth College), Margaret Karagas (Geisel School of Medicine at Dartmouth College), Hannah Laue (Geisel School of Medicine at Dartmouth College)

Background: Triclosan (TCS), an antimicrobial and endocrine-disrupting chemical, is used in many consumer products and may impact human health, including the microbiome. While certain uses of TCS were recently banned by the FDA, other uses remain unregulated. We aimed to investigate the effects of triclosan exposure during pregnancy on the infant microbiome.
Methods: We measured urinary TCS in samples from approximately the 12th to 13th week of pregnancy. The gut microbiome diversity and taxa of infants were analyzed using 16S rRNA V4-V5 sequencing. The study included 27 infants aged 6 weeks to 4 months and 13 infants aged 6 months to 1 year. Alpha diversity was assessed using Shannon’s Diversity Index. Associations between TCS and taxa abundance was evaluated using MaAsLin2. Models were adjusted for maternal age, maternal BMI, and mode of delivery.
Results: In early infancy, every 1% increase in pregnancy TCS was associated with a 0.010 unit increase in Barnesiella abundance and a 0.005 increase in Herbaspirillum abundance (p-value = 0.025, 0.031, respectively). Among older infants, every 1% increase in TCS related to a 0.025 increase in Barnesiella abundance, a 0.017 increase in Ignavibacterium abundance, a 0.009 increase in Vicingus abundance, and a 0.010 increase in Lachnoclostridium abundance (p-value = 0.030, 0.012, 0.008, 0.002, respectively). Maternal triclosan exposure appeared unrelated to the alpha diversity of the infant microbiome in either age group.
Conclusions: This study advances our understanding of the impacts of maternal triclosan exposure on the infant microbiome.
Impact: As one of the first studies to investigate the relationship between triclosan and the microbiome, this research provides a foundation for future studies. Further exploration of the link between maternal triclosan exposure and the formative period of the infant microbiome is crucial due to its potential lifelong impacts on human health.
Convergent Mechanisms of Nanobody-Mediated Neutralization of the CFTR Inhibitory Factor Cif
Adam R. Simard* (Dartmouth College), Noor M. Taher (Dartmouth College), Akaash K. Mishra (Dartmouth College), Kathryn S. Beauchemin (Dartmouth College), Natalia Vasylieva (University of California, Davis), Christophe Morisseau (University of California, Davis), Dean R. Madden (Dartmouth College).
Cif is a dimeric Pseudomonas aeruginosa epoxide hydrolase that inhibits airway clearance and facilitates more damaging airway infections in Cystic Fibrosis (CF) patients [1,2]. By diverting the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) to degradation pathways and destroying a pro-resolving signaling intermediate, Cif exacerbates CF lung pathophysiology while threatening to attenuate the benefit of therapies that restore CFTR-mediated airway homeostasis [2-4]. We are therefore pursuing strategies for the rapid detection and effective inhibition of Cif in therapeutic contexts. Previously, we developed a panel of high-affinity anti-Cif nanobodies and used them to establish an ELISA displacement assay to assess inhibitor potency [5]. Here, we present a mechanistic blueprint for this assay and investigate the structural relationships governing nanobody-mediated neutralization of Cif. We accomplished this by co-crystallizing Cif with a diverse subset of nanobodies and using X-ray crystallography to determine the structures of six different Cif:nanobody complexes as well as a tripartite structure of Cif simultaneously bound by two different nanobodies. A subset of Cif:nanobody structures share a preferred mechanism of neutralization where a bulky hydrophobic sidechain donated by complementarity determining region (CDR) 3 of the nanobody inserts into the Cif active site. Unexpectedly, this phenomenon is recapitulated by two nanobodies using CDR2 instead of CDR3 demonstrating convergent mechanisms of Cif neutralization. Following superposition by least-squares fitting of Cif co-crystallized with inhibitors or nanobodies, we conclude that the binding of inhibitors and nanobodies to Cif creates steric overlap, thereby validating the mechanistic basis for the ELISA displacement assay. Lastly, the tripartite structure highlights a pair of nanobodies that recognize non-overlapping Cif epitopes. This enabled the development of a cost effective and sensitive assay to quantitate native Cif in clinical samples, and permits exploration of intra- and inter-molecular conformational regulation of Cif activity.

1. Hvorecny KL, Dolben E, Moreau-Marquis S, Hampton TH, Shabaneh TB, Flitter BA, et al. An epoxide hydrolase secreted by Pseudomonas aeruginosa decreases mucociliary transport and hinders bacterial clearance from the lung. Am J Physiol Lung Cell Mol Physiol. 2018 01;314(1):L150–6.
2. Flitter BA, Hvorecny KL, Ono E, Eddens T, Yang J, Kwak DH, et al. Pseudomonas aeruginosa sabotages the generation of host proresolving lipid mediators. Proc Natl Acad Sci USA. 2017 03;114(1):136–41.
3. Bomberger JM, Ye S, MacEachran DP, Koeppen K, Barnaby RL, O’Toole GA, et al. A Pseudomonas aeruginosa Toxin that Hijacks the Host Ubiquitin Proteolytic System. PLoS Pathog [Internet]. 2011 Mar 24 [cited 2020 Jul 8];7(3).
4. Stanton BA, Coutermarsh B, Barnaby R, Hogan D. Pseudomonas aeruginosa Reduces VX-809 Stimulated F508del-CFTR Chloride Secretion by Airway Epithelial Cells. PLoS One. 2015 May 27;10(5):e0127742.
5. Vasylieva N, Kitamura S, Dong J, Barnych B, Hvorecny KL, Madden DR, et al. Nanobody-based binding assay for the discovery of potent inhibitors of CFTR inhibitory factor (Cif). Analytica Chimica Acta. 2019 Jan;S0003267019300273.
Fabrication of an in-vivo MRI-compatible knee loading device
Russell Perdue* (University of Delaware), Isabelle Larche (University of Delaware), Jamie Benson (University of Delaware), Alvin Su (Nemours Children’s Health), and Dawn Elliott (University of Delaware)

A common type of knee injury among youth is a torn meniscus. While the surgery for repairing a torn meniscus is relatively straightforward, there is no consensus on rehabilitation protocol post-surgery. The goal of this project was to create an in-vivo MRI-compatible device that (a) positions the knee at a range of angles and (b) applies a load to the knee between 0 and 1 body weight, so that meniscus and repair site mechanisms can be studied in vivo.  The base of the device sits within an inset of an MRI bed in place of an MRI head coil. Hinged to the base, there is plate that rotates between 0 and 90 degrees in increments of 10 degrees. Parallel to this plate, there are two plates that extend up to 2 inches using scissor jack mechanisms. The subject’s foot is strapped to the outermost of these plates. Between these plates, there is a load cell used to measure the applied load, and straps that secure the subject’s knee to the device.  The current design of the device addresses the challenges with the previous device, those being load dissipation during experimentation, uneven loading, poor ease of use, and the presence of non-MRI-compatible materials. This will be validated in the future with experimental testing, at which point the device can start to be used for meniscus research.
Mitochondrial CD36 expression is increased in endothelial cells exposed to elevated glucose in vitro
Erica J. Johnson* (University of Delaware), Dr. Ibra S. Fancher (University of Delaware), Thanh Nguyen (Mayo Clinic)

Obesity is a multifactorial disease encompassing several cardiovascular risk factors, including hyperglycemia. CD36 is a fatty acid translocase which was shown to be upregulated by obesity and hyperglycemia. Recent studies have identified the expression of CD36 in mitochondria of a variety of cell types which, following upregulation of mitoCD36 expression, impacts cellular respiration. Therefore, we aimed to determine the effect of hyperglycemia on mitochondrial CD36 expression in endothelial cells in vitro.
We treated human adipose microvascular endothelial cells (HAMECs) with high glucose (200 mg/dL) and mannitol (osmotic control; 200 mg/dL). Following treatments, CD36 expression was assessed via Western blot, qPCR, flow cytometry and immunocytochemistry. A CD36 antibody was used to identify membrane expression. Mitochondria were isolated from HAMECs using positive selection of TOM40 via magnetic associated cell sorting.
HAMECs exposed to elevated glucose exhibited an increase in CD36 expression in total cell lysates and in permeabilized cells after 72 hrs. No changes were detected in CD36 membrane expression or in CD36 gene expression following elevated glucose, suggesting that internal expression of CD36 protein is increased independent of transcription under these conditions. To determine if CD36 expression was elevated specifically in mitochondria following hyperglycemia, mitochondria were isolated and purified from HAMECs. CD36 expression was increased in the mitochondria of glucose treated cells, perhaps indicating alterations to mitochondrial respiration in the presence of sustained hyperglycemia.
Our findings reveal that mitochondrial CD36 expression is elevated in endothelial cells exposed to sustained increases in glucose in vitro, which may impact mitochondrial respiration in the presence of other macronutrients, like fatty acids. Ongoing studies in our lab are aimed at identifying the functional effects of increased mitoCD36 on endothelial cell respiration when challenged with fatty acids.
Disease-modifying Therapies for Sickle Cell Disease: Decisional Needs and Supports among Adolescents and Young Adults
Ke Ding, Ph.D.* (Nemours Children’s Health), Ben Bear, M.S. (Nemours Children’s Health), Erica Sood, Ph.D. (Nemours Children’s Health, Thomas Jefferson University), Melissa Alderfer, Ph.D. (Nemours Children’s Health, Thomas Jefferson University), Lori Crosby, Psy.D. (Cincinnati Children’s Hospital Medical Center, University of Cincinnati), Aimee K. Hildenbrand, Ph.D. (Nemours Children’s Health, Thomas Jefferson University)

Adolescents and young adults (AYAs) with sickle cell disease (SCD) often experience significant challenges with disease management. Hydroxyurea and several newer disease-modifying therapies have demonstrated health benefits; however, uptake of these therapies remains suboptimal. Shared decision-making is one promising solution to addressing barriers in uptake and to improve health outcomes for AYAs with SCD. A thorough understanding of AYAs’ decisional needs can guide the development of decisional supports and promote shared decision-making. Informed by the Ottawa Decision Support Framework, this study aimed to examine decisional needs and current supports reported by AYAs with SCD, their caregivers, and SCD healthcare providers. Data were obtained through qualitative interviews (14 AYAs, 11 caregivers) and online crowdsourcing (40 healthcare providers). Thematic and descriptive content analyses were used to summarize perspectives on decisional needs and supports. Adequate inter-rater reliability was achieved (Cohen’s Kappas > .80). Families reported needing six areas of support: resolving decisional conflicts, gaining knowledge, clarifying expectations, garnering supports and resources, navigating complexities in the decision, and addressing other personal concerns. Families reported receiving supports from providers that facilitated their decision-making by helping them: gain knowledge, clarify expectations, address other personal concerns, build rapport with providers, and resolve unreceptiveness or ambivalence to making a decision. Providers reported offering additional supports targeting: inadequate experience in implementing therapies (e.g., pill swallowing), inadequate health and social services and financial assistance, and inadequate instrumental support (e.g., transportation). This research highlights the needs AYAs and caregivers identify when making decisions about disease-modifying therapies for SCD and the decision supports offered by healthcare providers. Many decisional needs reported by families corresponded to supports offered by providers, though some needs appear to be unmet by current supports. Findings will inform the development of a shared decision-making intervention for AYAs with SCD, their caregivers, and healthcare providers.
Using Drosophila as a Model to Study Environmental Toxicity Relevant to Parkinson’s Disease
Angeline Claudia Atheby* (DSU), Lindsey Ruggiero (DSU) Hakeem Lawal (DSU)

Parkinson’s Disease (PD) is the second most common neurodegenerative disease characterized in part by the loss of dopaminergic neurons in the substantia nigra pars compacta. Epidemiological studies have established that exposure to certain pesticides and herbicides increase an individual’s likelihood of developing PD. Although the full catalog of pesticides that pose risks for PD is not currently known, some potentially toxic compounds are commonly used for agricultural or gardening purposes, and may be purchased at local stores. The objective of this study therefore, was to determine whether different combinations of these commercially-available pesticides and herbicides have either a synergistic or additive effect on toxicity in Drosophila. We first established a dose response curve in the following pesticides: acephate, atrazine, and diuron. We then exposed Drosophila to different combinations of the compounds at a concentration at which each alone was minimally toxic. We performed survival and locomotion ability analyses on each experimental group. Here, we report that combinations involving atrazine and diuron show strong decreases in survival that were greater than each single pesticide alone. We also present data on our analysis of the effects of these compounds on reactive oxygen species generation and measures of neurotoxicity. Together, our data suggest that exposure to combinations of commonly-used pesticides such as atrazine and diuron may elevate susceptibility to environment toxins.
F-Actin Bundles and Cross-linkers in Osteocyte Dendrites
Rosa Guerra* (University of Delaware), Megan Coffin(University of Delaware), Shannon Modla(University of Delaware), Velia Fowler(University of Delaware), Liyun Wang(University of Delaware)

Osteocytes are the master orchestrators of bone remodeling, and their extensive dendrite network enables them to sense the external mechanical signals and communication with other cells.1,2 With aging, the dendrites deteriorate with bone quality declined.3 Similar to epithelial microvilli and inner ear stereocilia4, osteocyte dendrites are thought to contain rigid tightly packed cross-linked F-actin bundles as in previous mechanosensing models.5 However, the dendrite cytoskeleton details have not been revealed experimentally due to their entombment in a mineralized bone matrix and the high resolution needed to resolve the F-actin fibers (5-7 nm diameter). The goal of this study is to optimize protocols for high-resolution STED imaging and transmission electron microscopy (TEM) of the dendrite cytoskeleton of cultured osteocytes.
MLO-Y4 cells were seeded at 4,000 cells/cm2 onto collagen-coated coverglass and cultured in alpha-MEM with 2.5% fetal bovine serum and 2.5% calf serum for 2-4 days. The F-actin cytoskeleton and selected cross-linkers were stained with SiR-Actin (in live cells), phalloidin and fimbrin and actinin antibodies (in 4% paraformaldehyde fixed cells), followed with STED (pixel ~16.5 nm) and confocal imaging (pixel ~100-200 nm). Additional TEM images (pixel ~0.55 nm) were also taken of the cells on coverslips.
In cultured osteocytes, there were a few long cellular protrusions containing linear or curved F-actin fibers in a semi-parallel fashion. These fibers were neither located at the center (under confocal) nor formed compact bundles (observed under STED and TEM). Under STED, fimbrin and actinin were found along the F-actin fibers and they were not fully co-localized with F-actin fibers in the dendrites as suggested in a previous study.6 Overall, the F-actin cytoskeleton of 2D cultured osteocytes is arranged differently than the previous linear actin core model. To understand osteocyte dendrites in vivo, better models are needed.
References: (1) Schaffler, Calcif Tissue Int, 2014 (2) Bonewald, JBMR, 2011 (3) Tiede-Lewis, AGING, 2017 (4) Pelaseyed, J Cell Sci, 2018 (5) Wang, PNAS, 2007 (6) Murshid, J Bone Miner Metab, 2007
The Influence of Extracellular Matrix Properties on Brain-Tropic Metastatic Breast Cancer Fate
Sam Freeman* (Univ of Delaware), John H. Slater (University of Delaware)

Tumor cell dormancy is regulated by many mechanisms including extracellular matrix (ECM) composition and degradability. Disseminated tumor cells (DTCs) from breast cancer can enter dormancy and remain there for months to decades before reactivation and new tumor development. Understanding how organ- specific ECM composition contributes to inducing dormancy is important for the potential development of new therapeutic approaches to prevent reactivation. Accordingly, we are investigating how organ-mimetic hydrogel formulations, with respect to differential integrin ligation, influences the dormancy process. To do so, we utilized two cell lines, the aggressive triple negative parental breast cancer cell line, MDA-MB-231 (231), and a brain-tropic sub line, MDA-MB-231-BrM2a (BrM2a). These cell lines were each encapsulated and cultured for 15 days in five poly(ethylene glycol) (PEG)-based hydrogel formulations composed of proteolytically degradable PEG, a generic or brain mimetic integrin ligating peptide formulation, and the non-degradable crosslinker N-vinyl pyrrolidone. Of these five formulations there are 3 classes of gels, a permissive formulation which is highly degradable and adhesive, a non-permissive formulation which is less degradable but still adhesive (NP1), and a non-permissive formulation which is highly degradable but non-adhesive (NP2). Comparing the results from the two permissive formulations demonstrated no peptide-specific influence on phenotype but a cell type dependence was observed with higher proliferation in the 231s in both the generic and brain-specific formulations. NP1 hydrogels indicated that the viability of BrM2a cells is supported by the brain-mimetic formulation with no significant difference observed in the 231s. NP2 hydrogels induced lower viability but higher proliferation in BrM2as compared to 231s again indicating a cell type dependence. These results demonstrate that the use of organ-mimetic, synthetic hydrogel formulations can better recapitulate the in vivo behavior of organotropic breast cancer compared to non-organ-specific generic formulations for modeling non-permissive environments.
Neuronal Autophagy Regulates Herpes Simplex Virus 1 Lifecycle Dependent on Autophagy Pathway Induction Mechanism
Paige Canova* (Dartmouth College), Hyung Suk Oh (Harvard University), David Knipe (Harvard University), David Leib (Dartmouth College)
Herpes simplex virus 1 (HSV-1) infects approximately two thirds of the world’s population and it establishes a lifelong infection through latency in the nervous system. Upon reactivation, the virus resumes lytic gene expression resulting in clinical manifestations such as cold sores, genital sores, ocular stromal keratitis, and encephalitis. One underexplored mechanism by which the host controls viral infections is through autophagy. Autophagy is a cellular mechanism used to recycle and remove misfolded proteins, damaged organelles, and intracellular pathogens. HSV-1 manipulates autophagy as a proviral mechanism to either enhance viral production or evade immune detection, but the role of autophagy in the control of HSV latency and reactivation is unknown. To examine this in a human in vitro model, we established a latency model in sensory neurons derived from inducible human pluripotent stem cells (iPSCs). Using iPSCs in conjunction with fluorescent viruses and a real time imaging system, we demonstrated that the impact of autophagy on the HSV-1 lifecycle is dependent on both the mechanism of autophagy induction, as well as the timing of autophagy induction relative to infection. HSV-1therefore likely utilizes multiple autophagy pathways to modulate latency and reactivation in response to different neuronal stress signals. Ongoing studies are elucidating the mechanisms of autophagy modulation that tilt the balance between lytic or latent gene expression. Current HSV-1 antiviral treatments only control replicating virus, but targeted manipulation of the autophagy pathway could allow control of HSV reactivation itself and lead to more effective treatments.
Changes in COVID-19 testing, vaccination, and preventative behaviors in Delaware’s underserved communities: An explorative study from a Longitudinal Survey 2021-2022
Kylie Trask*, Department of Neuroscience, Duke University
Xuanren Goodman, Department of Sociology and Criminal Justice, Delaware State University

Underserved communities, which often have a high proportion of residents identifying as Black or Latinx, have been disproportionately affected by Covid-19. From the beginning of the pandemic in March 2020 to the end in May 2023, policies and mandates were enacted by local, state, and national governments, resulting in varying attitudes and adherence to such interventions across communities. The present study investigates changes in COVID preventative measures among Delawareans, including vaccination, testing, and other protective behaviors from early 2021 to late 2022. We will also determine whether changes in COVID preventative measures differed by racial background. We use longitudinal surveys conducted in three waves between March 4th, 2021 to November 11, 2022. About half of the participants across the three waves identified as Black. Results show that Covid-19 vaccination increased from 43% to 62% to 70.6%, respectively, across the first, second, and third waves. In terms of engagement in preventative measures, there was a decrease in respondents that reported mask-wearing from 90% to 81% from wave one to wave two. During the collection of third-wave data, 79% of respondents reported mask-wearing when out in public. Across all waves, vaccination rates were lower among Black respondents compared to White respondents. However, Black respondents employed a higher level of preventative behaviors (i.e. hand washing and face mask wearing) compared to Whites. Scientific and policy implications will be discussed in the poster.
An integrative proteotranscriptomic approach identifies new substrates and downstream pathways for ADAM9 in colorectal cancer cells.
Congyu Lu (University of Delaware), Neha Sindhu*(University of Delaware), Jessica Rainey(University of Delaware), Xiaolu Xu(University of Delaware), Shawn W. Polson(University of Delaware), Jing Qiu(University of Delaware), Shuo Wei(University of Delaware)
ADAM9 (A disintegrin and a metalloprotease 9) is a single-pass type I transmembrane protein with proteolytic activity at its extracellular protease domain. Its overexpression is associated with several diseases including many solid tumors. For tumors, ADAM9 is reported to be associated with progression, aggressiveness, and metastasis. Juxtaposing with the fact that the loss of ADAM9 has little reported consequence aside from retinal degradation, the protein is potentially an excellent therapeutic target for multiple diseases including, but not limited to, solid tumors.

We use omics techniques to find the direct proteolytic substrates and indirect targets of ADAM9 in colorectal cancer cells. Transcriptome and Cellular Proteome of ADAM9 knockdown cells are integrated using the O2PLS technique. The integrated analysis of the differential gene and protein expression along with the direction in which they are corrected with ADAM9 differential expression is used to predict its transcriptional and post-transcriptional targets. Further, the post-transcriptional targets that are negatively correlated with ADAM9 which are also surface proteins are predicted to be candidate substrates that could be directly cleaved by ADAM9. Some of these in-silico predictions are further confirmed in our lab using western blotting.
Measuring microscale mechanical properties in synthetic hydrogels by combining experimentally measured microindentation induced hydrogel displacement fields with finite element modeling
Ryan Taitano* (University of Delaware), Omar Banda (University of Delaware), and John H. Slater (University of Delaware)

Due to their ease of tailorability, synthetic hydrogels have become a popular material used in investigating how extracellular matrix physical properties influence cell behavior. Because of this, characterizing hydrogel mechanical properties is of great importance. For soft hydrogels used in the context of cell-culture based applications (~1-30 kPa), performing standard tensile or compression testing is often infeasible. Indentation experiments are often used as an alternative, but they only measure the surface maximum indentation assuming Hertzian contact mechanics. Additionally, most microindentation experiments require an assumption that the hydrogel is incompressible and therefore Poisson’s ratio (v) is ~0.5.  In this study, we developed an approach to simultaneously estimate the Young’s modulus (E) and v via gravity-based microindentation experiments in poly(ethylene glycol) diacrylate (PEGDA) hydrogels. We photocoupled 3D arrays of fluorescent fiducial markers within PEGDA hydrogels using 2-photon laser scanning lithography (TP-LSL), and subsequently imaged the markers during microindentation using spheres of known diameters (150-700 um), to measure the 3D displacement fields. The 3D displacement fields were fit with finite element models (FEM) to estimate the hydrogel mechanical properties (E, v). Additionally, we employed a robust statistical approach, the Markov-Chain Monte Carlo (MCMC) method, to estimate the confidence bounds for the measured mechanical properties. Using this approach, we investigated the possibility that the micromechanical properties (E and v) change from the surface of the hydrogel deeper into the bulk hydrogel.
Our approach to generating a regularized grid array of fiducial markers within hydrogels combined with computational approaches to simultaneously measure both Young’s modulus and Poisson’s ratio provides a unique advantage in characterizing soft biomaterials and how changes in biomechanical properties alter cell behavior.
A Macrophage-specific Effect for Mycophenolate Mofetil in the Treatment of Systemic Sclerosis
Patrick Murphy* (Colby-Sawyer College), Em Morris (Dartmouth College), Patricia Pioli (Dartmouth college)

 The root cause of the autoimmune disease systemic sclerosis (SSc) is unknown. SSc causes life-threatening fibrosis in multiple tissues, including the skin, kidneys, lungs, and heart. Treatment of SSc patients with Mycophenolate Mofetil (MMF), an inhibitor of guanine nucleotide synthesis that targets the enzyme inosine monophosphate dehydrogenase (IMPDH), has been shown to relieve SSc-associated inflammation and fibrosis. Prior work has shown that MMF inhibits proliferation of B and T lymphocytes, which are white blood cells that mediate adaptive immune responses and require guanine synthesis to expand. While MMF is thought to exclusively target lymphocytes, the effect of this drug on other immune cells is unknown. An immune cell that has been implicated in our prior studies as regulators of fibrosis and inflammation in SSc are macrophages. We hypothesized that one potential mechanism by which MMF inhibits fibrosis in SSc is through direct effects on macrophages. To test this, monocytes from healthy human donors were cultured in the active metabolite of MMF, mycophenolic acid (MPA), and analyzed for effects on cell viability and caspase activity, as well as mRNA and surface marker expression and cytokine production. A subset of macrophages not killed by MPA showed attenuation of pro-inflammatory activation, as demonstrated by mRNA and surface marker expression, as a result of drug treatment. The addition of guanosine to cultures reversed the effect of MPA on macrophages, suggesting MPA-mediated changes in activation and viability are dependent on IMPDH. Collectively, these results implicate MMF as a modulator of macrophage viability and activation and suggest the clinical benefit of MMF in SSc may be mediated at least in part through effects on macrophages. Future work will utilize a novel 3D fabricated tissue model of SSc dermal fibrosis to test the effect of MPA on macrophage viability and activation in SSc skin.  
CD36 ablation restores endothelial function in mesenteric arteries of diet-induced obese mice.
Sabita Rokka, Emma Hudgins, Ibra S Fancher
Department of Kinesiology and Applied Physiology, College of Health Sciences, University of

The vascular endothelium is sensitive to obesity-induced stress resulting in endothelial dysfunction (ED), a precursor to cardiovascular disease. However, not all vasculature exhibits ED with obesity: visceral adipose arteries are impaired whereas subcutaneous adipose arteries retain endothelial function. We recently showed that impairment of the inwardly rectifying K+ channel, Kir2.1, in obesity mediates visceral adipose artery ED, however, the mechanisms governing Kir2.1 impairment remain unknown. Because Kir2.1 is inhibited by several intracellular fatty acid derivatives known to be elevated in obesity, we aimed to determine if ablation of the fatty acid translocase CD36 rescued endothelial function in visceral (mesenteric) arteries of obese mice. Mesenteric and subcutaneous adipose arteries were isolated from lean and obese WT or CD36-/- mice and flow-induced vasodilation was used to assess endothelial function ex vivo. Mesenteric arteries from obese WT mice exhibited ED as compared to lean counterparts and subcutaneous arteries. In contrast, mesenteric arteries from CD36-/- mice showed full recovery of the dilatory response to flow, an effect dependent on the functional recovery of Kir2.1. We next tested if distinct adipose depots mediate differential expression of endothelial Kir2.1 and CD36 in vitro. Primary adipose microvascular endothelial cells were incubated with subcutaneous or visceral adipose tissue from lean or obese mice. CD36 and Kir2.1 expression was assessed using qPCR and immunoblotting. Incubation with adipose did not induce changes in CD36 and Kir2.1 expression. Together, these findings point to an obesity-induced functional upregulation of CD36 as a key contributor to Kir2.1 impairment and ED in visceral adipose arteries.
Volumetric Changes Within a Two Hit Mouse Model of Neonatal Hypoxic Ischemic Encephalopathy
Victoria Langdon* (Nemours Children’s Health), Elise Lemanski (Nemours Children’s Health and University of Delaware), and Elizabeth Wright-Jin, MD, PhD (Nemours Children’s Health and University of Delaware).

With approximately 10,000 births per day, neonatal Hypoxic Ischemic Encephalopathy (HIE) affects 1.5 out of every 1000 infants and is the leading cause of cerebral palsy in children born at term, often resulting in long-term motor and cognitive deficits. The only currently available treatment is therapeutic hypothermia which must be applied within the first six hours of birth. Since maternal infection and fever are risk factors for HIE, our mouse model combines maternal immune activation through a peripheral LPS injection in late gestation (GD18) with hypoxia at P6 to better represent the injury seen in humans. In humans, there are patterns of injury that include deep gray matter damage in regions like the dorsal striatum and hippocampus, white matter damage, and watershed injury. This study explores volumetric differences of brain regions relevant to motor and cognitive deficits seen in human infants and animal models of HIE as a measure of injury. We conducted volumetric analysis of the dorsal striatum, dorsal hippocampal, ventral hippocampal, and whole brain regions through two methods: MRI and histological analysis. MRI brains were collected at P7 (neonates) and P75 (adults). Histological brains were collected at P7, P8, and P9 post hypoxia at P6 and stained with cresyl violet prior to imaging. While histological regional volumes were not different, ex vivo MRI results indicate a lower brain volume in neonatal (P7) HIE effected mice in the dorsal striatum and whole brain (p < 0.05). Adult (~P75) MRI analysis revealed that only males exposed to HIE have a volumetric decrease in the dorsal hippocampus (p < 0.05). The MRI volumetric changes are consistent with the deficits typically seen in HIE behavior in addition to volumetric decreases seen in human infants with HIE and point to a long-lasting sex-specific impact of HIE within our model.
Characterizing the effects of altered cholinergic synaptic transmission during aging
DaShan Osborne* (Delaware State University), Rohina Nemat (Delaware State University), Hakeem O. Lawal (Delaware State University).

Acetylcholine (ACh) plays a central role in the regulation of key life functions including locomotion and cognition. Once synthesized in the cytoplasm, ACh is transported to and stored in synaptic vesicles by the Vesicular Acetylcholine Transporter (VAChT). Despite a wealth of knowledge about the regulation of ACh synaptic transmission and its role in aging, much remains poorly understood regarding how cholinergic release is mediated late in the lifespan or its precise role in behavioral decline during aging. We are interested in determining how ACh synapses, and the behaviors they mediate, are altered during aging; and the role that changes in the expression or function of VAChT may play in that process. We seek to visualize age-related changes in the expression and localization of VAChT, and measure alterations in ACh-linked behaviors. We report that increases or decreases in VAChT function cause deficits in behaviors like locomotion that a regulated in part by ACh. We also show that alterations in VAChT impair a thigmotaxis-like behavior exhibited by flies. Moreover, we present preliminary reports on our study aimed at determining the subcellular localization of VAChT in aged flies. Future studies will focus on how changes in age and/or VAChT expression alter synaptic structure and physiology. Together, this work demonstrates the relevance of the central cholinergic signaling in the regulation of key behaviors during the lifespan.
Exploring the Interactions between Lytic Bacteriophages and Airway Epithelial Cells: Implications for Phage Therapy and Host Response
Braden C. Brassard* (Saint Anselm College), Paula F. Zamora (Dartmouth College), Jennifer M. Bomberger (Dartmouth College)

Phage therapy is emerging as a promising approach for treating multidrug-resistant (MDR) bacterial infections. While phages are considered safe and effective against bacteria, their interactions with the human host remain incompletely understood. In this study, we investigated the associations between lytic bacteriophages and airway epithelial cells. Given that bronchial epithelial cells serve as the primary target of biofilms within the airways of individuals infected with bacterial respiratory infections—such as Pseudomonas aeruginosa infections in patients with cystic fibrosis (CF)—they are anticipated to encounter the greatest phage exposure during phage therapy. Previous results from the laboratory demonstrated that the interaction between phages and airway epithelial cells depends on specific phage properties and the physicochemical environment. Although phage uptake by bronchial epithelial cells was limited, the cells detected phages through pattern recognition receptors (PRRs) and mounted a transcriptomic response enriched in immune genes. Furthermore, different phages induced distinct cytokine secretion profiles, indicating phage-specific immune responses. We aim to highlight the heterogeneity of cellular responses to phages and emphasize the importance of considering phage-mammalian interactions in phage therapy by further validating the involvement of the PRRs Toll-like receptor ½ (TLR½), 2 (TLR2), and 4 (TLR4), as well as Stimulator of Interferon Genes (STING), in phage-epithelial signaling. We established a model using known agonists and antagonists of these signaling pathways to which subsequent testing with phages will be compared in order to confirm the previous results that the immune response varies with different phages. Assessing these interactions during phage characterization could improve the safety and efficacy of phage-based treatments. The study expands our understanding of phage therapy and its effects on the host, providing insights into the complex interplay between phages, bacteria, and the human immune system.
Simultaneous TMS and fMRI for Cortical Inhibition of the Long-Latency Response in the Forearm
Cody A. Helm* (University of Delaware), Sheldon-Anthony Amofah (University of Delaware), Rebecca C. Nikonowicz (University of Delaware), Fabrizio Sergi (University of Delaware)

Reticulospinal tract (RST) knowledge is limited due to a lack of methodologies to directly quantify RST function [1]. Our lab has developed StretchfMRI [2] to quantify activity in the brainstem associated with a long-latency response (LLR), however it cannot decouple the contribution from the RST with the corticospinal tract (CST). TMS can decouple the contribution of the CST to the LLR [3], however TMS and fMRI have not yet been combined to study the causal role of the RST on the LLR. The overall objective of this project is combine TMS with fMRI to determine the causal role of the RST on the LLR in the forearm. For experiment 1, we combined surface electromyography (EMG), wrist perturbations, and subthreshold TMS to determine the effect of TMS intensity and timing on the LLR amplitude (LLRa) of the flexor carpi radialis (FCR). We tested two intensities (90% and 95% AMT) and three TMS timing levels (0, 20, and 50 ms prior to perturbation onset). For experiment 2, we combined surface EMG with an MRI-compatible TMS coil to quantify the effect of TMS on whole brain BOLD signal during isometric contractions of FCR. We tested the effect of subthreshold TMS with two modes, TMS and no TMS. In experiment 1, we found a significant effect of TMS timing on the LLRa (p<0.0001). From post-hoc analysis, 20 and 50 ms timings significantly reduced the LLRa compared to the control mode, PertOnly (p=0.0231; p<0.0001, respectively). For experiment 2, we found a significant increase in BOLD signal associated with TMS in the left motor cortex, the sensory cortex, and the auditory cortex. In conclusion, the ability of subthreshold TMS to reduce the LLRa significantly depends on TMS timing. Subthreshold TMS can modulate the BOLD signal in the primary motor cortex resulting in later cortical inhibition.
Liquid Metal Strain Sensors For Pediatric Patients
Claudia McCormick* (Univ of Delaware), Nathan Lazarus (Univ of Delaware)

Juvenile Arthritis (JIA) affects 1 in every 1,000 children in the US. Current medical devices for JIA patients’ are largely designed for and tested on adults, with slight modifications for pediatrics. These devices fail to adapt to a child’s anatomy, which can lead to misdiagnosis. Stretchable sensors are a common wearable device in biomechanics, used to measure volume expansion and motion changes in patients’ joints. These sensors can be used longitudinally as a child’s body develops. This project aims to understand how scaling down a stretchable strain sensor affects its properties. In this design, four stretchable strain sensors were developed that include an embedded channel of non-toxic liquid metal, eutectic gallium indium (EGaIn). The channel length of these sensors are 50 mm, 40 mm, 30 mm, and 20 mm. To understand how to scale down the sensors, we must understand how different sized channels vary during longitudinal studies. Therefore, the resistance values during loading were recorded up to 120% strain. Scaling down the channels reduced the resistance in each sensor; the 50 mm channel has a resistance of 2.593 ohms and the 20 mm channel has a resistance of 1.455 ohms at 120%. The sensitivity changes during scaling were also analyzed, which is done by calculating the rate of change in resistance of the channel sizes. These calculations showed a noticeable difference; the 40 mm channel had the greatest rate of change in resistance followed by the 50 mm channel, 30 mm channel, and lastly the 20 mm channel. Overall, for the sensors to be used longitudinally, it is important to understand how the properties of the sensor change when it is scaled. Based on these results, we can customize these sensors for a broad range of sizes to adapt to a child’s anatomy.
Blood vessel formation controlled by selective endothelial cell sorting
Bruno Esmeraldo*(University of Delaware), Tom Lee(University of Delaware), Brian Kwee(University of Delaware)

Vascularization of biomaterials with implanted cells has been regarded as a key challenge in engineering tissues for regenerative medicine, as blood vessels are required for delivering nutrients to implanted cells. The process of recruiting the host’s blood vessels by angiogenesis to the biomaterial is slow, and the time lag between biomaterial implantation and angiogenesis often results in cell death of implanted cells due to the initial lack of nutrient transfer to the cells. Therefore, in vitro pre-vascularization of biomaterials with endothelial cells (blood vessel cells) has been explored as a strategy for enhancing the viability of implanted cells. However, the underlying factors that contribute to de novo vasculogenesis, the self-assembly of endothelial cells into a vessel network in biomaterials, are still unclear. Understanding the role of receptors that mediate cell-to-cell, and cell-to-extracellular matrix (ECM) interactions, such as β1 integrin and VE-Cadherin, and the expression of these receptors is paramount to the formation of robust blood vessel networks. In the present work, we utilized fluorescence-activated cell sorting to sort human umbilical vein endothelial cells into four subpopulations with different combinations of “high” or “low” β1 integrin or VE-Cadherin expression. Vasculogenesis of the subpopulations was then evaluated in collagen and fibrin hydrogels, after which blood vessel network formation was measured by fluorescently staining the cells and quantifying lumen formation, number of branching vessels, and interconnectivity of the vessel networks. Initial data suggests that the “Low” VE-Cadherin/“High” β1 integrin cell group demonstrated the greatest degree of vascularization among the subpopulations. This initial result suggests that the “Low” VE-Cadherin/“High” β1 integrin group may have the greatest ability to pre-vascularize implantable biomaterials, to enhance the survival of implanted cells..
Deconstructing a Hot Spot of Advanced Breast Cancer Among Women in Wilmington: An Exploratory Study on Root Causes
Atif Bacchus* (INBRE, The University of Delaware, ChristianaCare, Helen F. Graham Cancer Center & Research Institute), Ross Budziszewski (ChristianaCare, Helen F. Graham Cancer Center & Research Institute), Yuchen Zhang(ChristianaCare, Helen F. Graham Cancer Center & Research Institute), Scott Siegel (ChristianaCare, Helen F. Graham Cancer Center & Research Institute)

Background: Preliminary research from the Helen F. Graham Cancer Center & Research Institute (HFGCCRI) identified an area in Wilmington as a ‘hotspot,’ or a geographical location with higher-than-average rates, for advanced breast cancer. The present study aims to understand the relationship between screening, clinical factors, and demographic characteristics and stage at diagnosis in the Wilmington hotspot.
Methods: Patient records (N = 68; 96% Black) were reviewed from the HFGCCRI Cancer Registry who resided in the Wilmington hotspot. Clinical, family, and social history, mode of diagnosis (breast cancer detected via clinical/symptomatic presentation or screening) and primary care engagement data were abstracted from patient’s electronic health records. Descriptive statistics and chi-square analyses were conducted, and two groups were compared: advanced (Stage 2a and above; n = 38) and non-advanced (Stage 1; n = 30).
Results: The advanced (Mage = 58.9, SDage = 13.5; 97% Black) and non-advanced (Mage = 64.6, SDage = 14.8; 93% Black) groups were not significantly different for variables including age, race, and BMI. The advanced group was significantly more likely to detect cancer clinically/symptomatically (57%) compared to the non-advanced group (33%): χ 2 (2, N = 68) = 6.625, p = 0.036. Compared to the non-advanced stage group, the advanced stage group had lower screening history (46% vs. 67%) and primary care engagement (22% vs. 37%). Non-significant trends in higher alcohol use for advanced patients compared to non-advanced patients (44% vs. 30%).
Conclusion: Preliminary findings suggest that patients with advanced breast cancer in the Wilmington hotspot may not be receiving mammograms as frequently or could possibly be experiencing barriers to access preventative care. Future work should aim to develop interventions to increase primary care engagement and educate patients about screening mammogram guidelines which may decrease the number of patients being diagnosed with advanced stage breast cancer.
Non-canonical inhibitors target CAL binding: new approaches to enhance mutant CFTR activity
Nicholas Gill* (Dartmouth), Pat Cushing (Dartmouth), Yu Zhao (Dartmouth), Luke Wallace (Dartmouth), Alex Pletnev (Dartmouth), Mark Spaller (Dartmouth), Sierra Cullati (Dartmouth), Scott Gerber (Dartmouth), Prisca Boisguerin (University of Montpellier, France), Dominick Casalena (Novartis), Doug Auld (Novartis), Siyu Wang (Duke), Bruce Donald (Duke), Dean Madden (Dartmouth)

Significant achievements have been made in the past decade towards correcting the fold of F508del-CFTR and enhancing its activity at the apical membrane. Despite this success, there are still no FDA-approved therapeutics that directly target the post-endocytic stability of F508del-CFTR. Upon internalization, it is rapidly degraded through binding the CFTR-associated ligand (CAL) PDZ domain (CALP), which triggers its degradation in the lysosome. Our group and others have shown that inhibiting CAL rescues F508del-CFTR at the cell-membrane and significantly enhances its channel activity, suggesting that ‘stabilizer’ compounds, especially in conjunction with current CF therapeutics, would augment treatment efficacy.  Unfortunately, previous efforts to design inhibitors of the CALP yielded high specificity peptides with weak affinity and a low potency small molecule inhibitor that covalently modifies Cys319 of CALP. To enhance target affinity without sacrificing specificity, we designed peptidomimetic inhibitors containing modified lysine scaffolds that were able to bind distal regions outside of the conserved peptide-binding cleft. We show by SAR that targeting the β1-β2 loop confers enhanced affinity for CALP while conserving specificity. We also identified a potent Cys319 modifier that modulates both the thermostability and peptide-binding affinity of CALP through a multi-step reaction involving multiple adducted states. Interestingly, the inhibitory effect is mediated through a non-covalent, allosteric interaction between two adducted CALP states. These data provide insight into CALP protein dynamic and offer a new strategy for designing CALP inhibitors.
New conformations of the hepatitis B virus capsid protein revealed by all-atom molecular dynamics simulations.
Carolina Perez Segura* (University of Delaware), Rasanjali L. Ranawaka (University of Delaware), Boon Chong Goh (Antimacrobial Resistance Interciplininary Research Group, Singapore- Massachusetts Institute of Technology Alliance for Research and Technology Centre), Jodi A. Hadden-Perilla (University of Delaware).

Hepatitis B virus (HBV) infection is a major global health concern, with over 250 million individuals affected worldwide. The viral capsid, composed of 120 core protein (Cp) homodimers, plays a crucial role in various stages of the HBV life cycle. Although the capsid is structurally well-characterized, having been widely investigated with X-ray crystallography and cryo-electron microscopy, all-atom
molecular dynamics (MD) simulations are revealing new Cp conformations and
quaternary relationships not yet captured by experimental methods. Here, we
present new insights into the conformational dynamics of the Cp spike and linker
domains, the quasi-sixfold capsid pore, and the capsid inter-dimer interface.
Our results are based on sampling the motion of the intact HBV capsid on the
microsecond timescale, in MD simulations that encompass six million atoms. The
new conformations we observe, along with characterizations of their likelihood to occur at equilibrium, are relevant to establishing the mechanisms of capsid function, as well as developing new capsid targeting antivirals.
Optimization of Scaffold Free Generation of Bioengineered Cartilage from Passaged Human Chondrocytes
Stephanie Richardson-Solorzano* (University of Delaware), and Justin Parreno (University of Delaware)

Articular cartilage is a specialized connective tissue found at the ends of bones that allows for frictionless movement of one joint surface over the other. Cartilage is made up of cells (chondrocytes) that reside in an extracellular matrix (ECM) mainly composed of proteoglycans (mostly aggrecan; Acan), collagens (mainly collagen type 2; Col2), and water. When damaged articular cartilage is incapable of repair.
A current gold standard therapy for repair of small focal defects is Autologous Chondrocyte Implantation (ACI). In ACI, a small portion of healthy cartilage is surgically removed, and chondrocytes are isolated, expanded, and reimplanted into the defect. Expansion of chondrocytes for cell number in monolayer culture results in chondrocyte dedifferentiation leading to reduced chondrogenic gene expression, Col2 and Acan increased fibroblast matrix expression (collagen I and tenascin C)
To stimulate cartilage matrix deposition by expanded chondrocytes, we developed a screening approach to examine the effects of a curated list of 23 pharmacological agents thought to promote chondrogenic differentiation or matrix deposition. In this screen, we seeded cells in 2D within a 96 well plate and treated the cells with each agent. After six days the cells were fixed and stained for cartilage and fibroblastic matrix molecules. We then tested the shortlist of agents on the stimulation of matrix production by human passage cells cultured in 3D. Cells cultured in 3D were exposed to agents for a period of twenty days and examined the effects on matrix thickness and the type of matrix deposited by RT-PCR, gene expression and tissue sectioning followed by immunofluorescence microscopy. Overall, four pharmacological agents appeared to be promising candidates to stimulate matrix deposition in passaged cells both grown in monolayer and 3D culture. Developing methodologies to redifferentiate passaged chondrocytes and improve cartilage matrix deposition will lead to more effective cell-based therapies for cartilage repair.
Transcriptional Profiling of Inflammatory Cytokines in a Novel Mouse Model of Hypoxic Ischemic Encephalopathy
Sudha Anilkumar* (Nemours Children’s Hospital, University of Delaware), Bailey Collins (Nemours Children’s Hospital, University of Delaware), Elizabeth Wright-Jin, MD-PhD (Nemours Children’s Hospital, University of Delaware)

Occurring in 1.5 per 1000 live births, hypoxic ischemic encephalopathy (HIE) is a leading cause of death and long term disability in neonates. HIE is denoted by prolonged lack of oxygen to the brain, triggering an inflammatory response mediated by microglia, the primary immune cells of the central nervous system. Neonatal HIE is heavily linked with maternal immune activation: newborns exposed to maternal infection during gestation are 8 times more likely to be diagnosed with HIE. At present, standard animal models are inconsistent with clinical HIE with respect to patterns of brain injury and adult outcomes. Our novel two-hit mouse model of HIE first exposes the fetus to maternal immune activation via lipopolysaccharide injections in late gestation followed by hypoxia at postnatal (P) day 6 to better mimic the pathophysiology of the condition.

In this study, the inflammatory phenotype of our HIE model was characterized at two time points (P7 and P14) via transcriptional analysis. Bulk RNA sequencing of microglia at P14 was performed on HIE and saline/normoxia controls. Gene set enrichment analysis revealed upregulation of TNF signaling via NF-kB in HIE microglia, indicating activation of pro-inflammatory cytokines within the model. Real time qPCR was conducted on brain hemispheres at P7 and P14 to establish system-wide expression patterns for selected pro and anti inflammatory cytokines. At P7, one day post hypoxic injury, pro and anti inflammatory cytokines were upregulated in comparison to the saline/normoxia control, representing the crosstalk between inflammatory signaling typical in HIE. By P14, both types of cytokines were downregulated, demonstrative of a recovery phase. Identified cytokine trends were comparable to those found in neonatal HIE patients, validating the clinical relevance of our model. Therefore, the study signifies the scope for the model’s utility in elucidating the disease mechanism and assessing possible remediations.
Passaged Articular Chondrocytes from the Superficial Zone and Deep Zone can be Redifferentiated to Regain Zone-Specific Properties
Thomas J. Manzoni* (University of Delaware), Elizabeth Delve (Lunenfeld-Tanenbaum Research Institute), Vanessa J Bianchi (Lunenfeld-Tanenbaum Research Institute), Joanna F. Weber (Toronto Metropolitan University), Suresh C. Regmi (University of Calgary), Stephen D. Waldman (Toronto Metropolitan University), Tannin A. Schmidt (University of Connecticut Health Center), Alvin W. Su (Nemours Alfred I. duPont Hospital for Children), Rita A. Kandel (Lunenfeld-Tanenbaum Research Institute), Justin Parreno (University of Delaware)

Purpose/Objective: Bioengineered cartilage is a promising therapeutic to heal cartilage defects. Bioengineered cartilage matrix must be rich in collagen type II (COL2) and aggrecan (ACAN), and be mechanically competent, withstanding compressive and shearing forces. One strategy is utilizing passaged full-thickness chondrocytes to generate repair tissue. Biomechanical properties in native articular cartilage depend on the zonal architecture consisting of three zones, superficial (SZ), middle (MZ), and deep (DZ). In articular cartilage the SZ produces lubricating proteoglycan-4 (PRG4), while DZ chondrocytes produce collagen type X (COLX) for integration into the subchondral bone. Zonal and chondrogenic expression is lost after cell number expansion. Here, we test the hypothesis that both SZ and DZ chondrocytes can form bioengineered cartilage rich in COL2 and ACAN, while only SZ-derived cells will express SZ-specific PRG4, and only DZ-derived cells will express DZ-specific COLX.
Methodology: SZ and DZ chondrocytes were isolated from bovine joints, and zonal subpopulations were separately expanded in 2D culture. At Passage 2, SZ and DZ chondrocytes were seeded, separately, in scaffold-free 3D culture within agarose wells and cultured in redifferentiation media.
Results: Monolayer expansion results in loss of expression for PRG4 and COLX in passaged SZ and DZ cells, respectively. By passage 2, both had similar expression for dedifferentiated molecule collagen type I (COL1). Redifferentiation of both SZ and DZ chondrocytes led to the expression of COL2 and ACAN in both passaged cell populations. However, only redifferentiated DZ chondrocytes expressed COLX, and only redifferentiated SZ expressed and secreted PRG4.
Conclusion/Significance: Zonal properties are only expressed by the original zonal cells after redifferentiation. To form bioengineered cartilage rich in COL2, ACAN, and COLX, while secreting PRG4, cultures must contain both SZ and DZ chondrocytes. Our ongoing work aims to develop strategies to stratify zonal chondrocytes in order to generate bioengineered cartilage with a zonal architecture.
Early Life Arsenic Exposure, Metabolomics, and Gestational Diabetes Mellitus
Ahlam Abuawad* (Geisel School of Medicine at Dartmouth; Columbia University Mailman School of Public Health), Anne Hoen (Geisel School of Medicine at Dartmouth), David Kirchner (University of North Carolina at Chapel Hill), Susan Sumner (University of North Carolina at Chapel Hill), Ana Navas-Acien (Columbia University Mailman School of Public Health), and Mary V. Gamble (Columbia University Mailman School of Public Health), and Margaret Karagas (Geisel School of Medicine at Dartmouth).

Background. Exposure to arsenic (As) in drinking water is associated with many adverse health outcomes such as gestational diabetes mellitus (GDM). Ingested inorganic arsenic (InAs) is methylated to mono-(MMAs) and di-methylated arsenic species (DMAs). One-carbon metabolism (OCM) is a biochemical process involved in As methylation that depends on micronutrients (e.g., choline). A study of three Bangladeshi cohorts found an inverse association between %MMAs and BMI was attenuated following adjustment for plasma choline.
Objectives. To evaluate if the association between urinary As exposure/methylation and GDM is mediated by OCM metabolites.
Methods. We leveraged data from participants of the New Hampshire Birth Cohort at 24-28 weeks-gestation, including maternal urinary As species, OCM metabolites, and GDM (n=410). Linear and logistic regression models were used to investigate the associations between As exposure/species and OCM metabolites, and between OCM metabolites and glucose intolerance/GDM, respectively. Four-way decomposition mediation analyses were used to evaluate if OCM metabolites mediated the association between As exposure/methylation and GDM.
Results. Total As (μg/L), %InAs, %MMAs, and %DMAs were 4.7, 10.9, 9.1, and 79.9, amongst participants with normal glucose levels, and 6.5, 12.5, 9.2, and 78.3 amongst participants with glucose intolerance/GDM, respectively. Total As was negatively associated with glycine, while As species were positively associated with several OCM metabolites: both %InAs and %DMAs with phosphatidylcholines (PC) such as lysoPC(16:0) and PC aa 32:1, and %MMAs with glutamate and valine. Glutamate, lysoPC(16:0), and PC aa 32:1 were positively associated with occurrence of glucose intolerance/GDM. Mediation of the association between %InAs and GDM by acylcarnitine was observed in participants with overweight and obese BMIs.
Conclusions. These results suggest that OCM-related nutrients may be involved in the association between As exposure/methylation and diabetes-related outcomes. This research may contribute to the weight of evidence used to inform effective policies to reduce As exposure and improve health.
Regulation of Tenocyte Gene Expression through Actin Binding Myocardin-related Transcription Factors
Valerie West (Univ of Delaware), Kameron Inguito* (Univ of Delaware), Karl Matthew Ebron (Univ of Delaware), Justin Parreno (Univ of Delaware)

Tendons are fibrous tissue that allows for the transfer of forces from muscle onto bones to enable movement. Tendinosis is a condition where the tendon’s collagen deteriorates and can be attributed to due to excessive loading onto the tendon. At the cellular level, tissue overload counterintuitively causes the under-stimulation of tenocytes. Tenocyte under-stimulation alters tenocyte gene expression levels by decreasing tenogenic mRNA levels while increasing both chondrogenic and protease mRNA levels. Concomitant with these gene expression changes are alterations in tenocyte filamentous (F-) actin, whereby cells depolymerize F-actin into monomeric, globular (G-) actin units. In this study, we hypothesized that actin depolymerization regulates gene expression through an actin-biding transcription factor, myocardin-related transcription factor (MRTF), which has a high affinity toward G-actin-binding.
To test our hypothesis, we exposed primary isolated tenocytes to Latrunculin A which prevents F-actin polymerization by sequestering G-actin. This led to an overall decrease in F/G-actin. We found that latrunculin treatment results in nuclear export of MRTF coinciding with decreases in tenogenic genes (collagen-1, scleraxis, and a-smooth muscle actin) and increases in the expression of chondrogenic (Sox9) and proteases (Mmp-3 and Mmp-13). To elucidate which genes were regulated by MRTF, we exposed tenocytes to an MRTF inhibitor, CCG1423, which prevents nuclear accumulation of MRTF. CCG1423 led to significant decreases in tenogenic genes with no effects on chondrogenic or protease genes.
In conclusion, actin depolymerization is a regulator of gene expression in tenocytes, partially through the regulation of MRTF. Ultimately, a greater understanding of the regulation of gene expression during tendinosis by actin may allow for new therapeutic opportunities against disease progression.
The Kegel Exercise and Athletic Incontinence Among Active Women
Megan Welch, RN*(Dartmouth-Hitchcock Medical Center), Bridget Linehan, PhD, APRN(Dartmouth-Hitchcock Medical Center)

Background: Up to 70% of women experience urinary incontinence (UI). Kegel exercises are one of the most commonly known treatments for UI, which involve contraction and relaxation of the pelvic floor muscles. While Kegel exercises are often recommended, patient education regarding the precise technique is generally absent, and many women teach themselves how to do Kegels, but incorrect technique is unproductive. Pelvic Floor Physical Therapists (PFPTs) recognize that Kegels do not fully treat UI in isolation. A broader approach is a more effective.
Objective: The purpose of this study is to explore the relationship between self-reported participation and medical verification of the Kegel exercise and subsequent rates of athletic incontinence among active women.
Methods: The Active Women’s Incontinence Screening Tool (AWIST) was administered to 209 females. Basic statistics were employed, and a single-factor ANOVA analysis was done.
Results: 44% of participants who participate in Kegel exercises experience athletic incontinence, compared to 35% of participants who do not perform Kegel exercises (P=0.19). 47% of participants who’ve verified with a medical provider that they are doing Kegels correctly experience athletic incontinence, compared to 38% of those who have never received verification (P=0.34).
Discussion: Contrary to the widely-held belief in the efficacy of Kegel exercises, this study demonstrates that participants who perform Kegel exercises don’t have lower rates of athletic UI compared to those who do not perform Kegel exercises, even if their technique is verified. Rather than continued focus on Kegel exercises, a broader approach, with guidance from a PFPT may lead to better outcomes.
Reducing cannibalism in an emerging waste upcycler
Gabriel Dasilva* (University of Delaware), Destiny Mann (University of Delaware), Micheal Crossley (University of Delaware)

Roughly half of US land is devoted to agriculture, yet one third of the food produced ends up in landfills. This food waste could be recycled by insects which could be returned to the food system, a process termed “waste upcycling”. The lesser mealworm (Alphitobius diaperinus) has emerged as a potentially important waste upcycler, but its efficiency is challenged by their tendency to cannibalize.  Here, we test the importance of three factors, ratio of food to larvae, larval density, and moisture availability, for mealworm mortality and biomass generation using controlled feeding bioassays.
The Effect of Digital/Virtual Motor Interventions on Motor, Social, and Cognitive Outcomes in Individuals with Various Developmental Disorders with and without Intellectual Disability: A Systematic Review
Jung-mei Tsai* (Univ of Delaware), Jacob Corey (Univ of Delaware), Ashwin Mhadeshwar (Univ of Delaware), Sudha Srinivasan (Univ of Connecticut), and Anjana Bhat(Univ of Delaware)

Individuals with various developmental disabilities (DD) display co-occurring motor, social communication, and cognitive impairments affecting overall functioning. Digital technologies (e.g., exergaming, virtual reality, videoconferencing, etc.) have been used increasingly to promote better outcomes. However, their efficacy in improving multisystem outcomes is not well-known. We conducted a systematic review to comprehensively examine the efficacy of digital motor interventions in individuals with various DD. We searched four databases: PubMed, PsycInfo, SCOPUS, and CINAHLfor the studies that used interactive technologies to offer movement therapies and reported other domain outcomes including cognitive, sensory/perceptual, and social-behavioral. The population of interest included individuals with autism spectrum disorder (ASD), attention deficit hyperactivity disorder (ADHD), Down syndrome (DS), and intellectual disabilities (ID). Our search resulted in 2682 articles of which 17 met criteria. Our findings indicated medium-to-large intervention effects for executive functioning, fine motor, gross motor, sensory/perceptual, and social communication skills and thus digital motor interventions are an effective, low-cost tool to extend the reach of clinicians to improve multisystem outcomes. We recommend exergaming and video conferencing as a tool to promote multisystem development in individuals with various DD.
Spontaneous Neuronal Activity in Cerebellum and Thalamus of Spinal Muscular Atrophy Mice
Morgan Dowling*, Jianli Sun (Delaware State University)

Spinal Muscular Atrophy, more commonly known as SMA, is a neurodegenerative disease affecting the motor neurons in the spinal cord. This degeneration predominantly affects infants resulting in muscle weakness and atrophy. The mechanism of SMA is not well understood. Our previous data have shown decreased spontaneous activity and neural degeneration in post-symptomatic cerebellum in SMNΔ7 mouse model. To study the pre-symptomatic changes in the mouse brain, we examined the spontaneous activity of thalamus and cerebellum in postnatal day 3 mice. This can tell us whether the cerebellum contributes to the pathology of the disease, or if the reduction of cerebellum activity is a secondary effect of the disease pathology. The thalamus and cerebellum from both control and SMA mice were sliced using a vibratome. These slices were placed on a microelectrode array in order to record activity. The results show the cerebellum showing the most frequency with high variation. We are unable to draw a solid conclusion from these data points.
This research is supported by NINDS grant #R25NS095371, #R15NS120154
Transfection of Breast Cancer Cell Lines with CD44-Intracytoplasmic Domain Deletion Mutant Constructs for Studies of the CD44-ICD-Mediated Signaling Pathway
Maurio Goings*, Esther Daniel and Karl E. Miletti-Gonzalez (Delaware State University)

The CD44 intracytoplasmic domain (CD44-ICD) is a 74 residues peptide generated by the proteolytic cleavage of CD44, a cell membrane receptor and adhesion molecule. This peptide can be translocated to the nucleus where we have detected via proximity ligation assays (PLA) a protein-protein interaction (PPI) between the CD44-ICD and the transcription factor Runx2 on the MMP-9 promoter region in MCF-7/CD44 cells. This PPI promotes the expression of the MMP-9 gene. The purpose of this study is to determine whether the CD44-ICD-RUNX2 complex can be interfered with using deletion mutants of the CD44-ICD. We hypothesize that these deletion mutants will block the CD44-ICD-Runx2 PPI complex and thus interfere with this signaling pathway. The CD44-ICD wild-type (wt) and seven deletion mutants were cloned in a Flag-tagging plasmid vector and each one of these constructs were transfected in MCF-7/pLXIN, a CD44 negative cell line, and in MCF-7/CD44 cells, a CD44 positive cell line. Utilizing cell lysates created from these transfections, Western Blotting analyses and Zymography techniques were used to assess the expression of CD44 and MMP-9, which are known genes to be in part regulated by the CD44 signal transduction pathway.
Volumetric Changes Within a Two Hit Mouse Model of Neonatal Hypoxic Ischemic Encephalopathy
Victoria Langdon* (Nemours Children’s Health), Elise Lemanski (Nemours Children’s Health and University of Delaware), and Elizabeth Wright-Jin, MD, PhD (Nemours Children’s Health and University of Delaware)

With approximately 10,000 births per day, neonatal Hypoxic Ischemic Encephalopathy (HIE) affects 1.5 out of every 1000 infants and is the leading cause of cerebral palsy in children born at term, often resulting in long-term motor and cognitive deficits. The only currently available treatment is therapeutic hypothermia which must be applied within the first six hours of birth. Since maternal infection and fever are risk factors for HIE, our mouse model combines maternal immune activation through a peripheral LPS injection in late gestation (GD18) with hypoxia at P6 to better represent the injury seen in humans. In humans, there are patterns of injury that include deep gray matter damage in regions like the dorsal striatum and hippocampus, white matter damage, and watershed injury. This study explores volumetric differences of brain regions relevant to motor and cognitive deficits seen in human infants and animal models of HIE as a measure of injury. We conducted volumetric analysis of the dorsal striatum, dorsal hippocampal, ventral hippocampal, and whole brain regions through two methods: MRI and histological analysis. MRI brains were collected at P7 (neonates) and P75 (adults). Histological brains were collected at P7, P8, and P9 post hypoxia at P6 and stained with cresyl violet prior to imaging. While histological regional volumes were not different, ex vivo MRI results indicate a lower brain volume in neonatal (P7) HIE effected mice in the dorsal striatum and whole brain (p < 0.05). Adult (~P75) MRI analysis revealed that only males exposed to HIE have a volumetric decrease in the dorsal hippocampus (p < 0.05). The MRI volumetric changes are consistent with the deficits typically seen in HIE behavior in addition to volumetric decreases seen in human infants with HIE and point to a long-lasting sex-specific impact of HIE within our model.
Social Density and Feeding Behavior in Drosophila melanogaster
Paul Schultz* (University of Delaware), Juntian Wei (University of Delaware), Lisha Shao (University of Delaware)

Socialization is essential. While human ecology is a social ecology, the effects of how extreme social density impacts an individual’s well-being remains unclear. Here we used Drosophila melanogaster, the common fruit fly, as a model organism to answer this question. We hypothesize that social isolation may cause negative outcomes by inducing stress pathways that intersect with essential behavioral and physiological functions. We tested the hypothesis by investigating the feeding behavior, which is regulated by both the animal’s internal state and the environmental factors. To empirically determine feeding behavior, the fly was fed dyed food for 24 hours. We found that socially isolated female virgin fruit flies ate significantly less than group-housed flies. To reveal the mechanisms underlying the altered behavior, we first defined the necessary length of time for social conditioning and found that acute (3-day) and chronic social (5-day) isolation may have distinctive regulation in fly’s feeding behaviors. Furthermore, we tested the reversibility of the behavior change. Our results showed that the change in feeding behavior after chronic isolation was reversed after 3-day regrouping, suggesting plasticity in fly’s reaction and adaptation to social densities. Importantly, we found that key metabolites were decreased in socially isolated flies. Further investigation on the causal relationship between metabolic and behavioral changes will provide more mechanistic insights into the regulation of feeding by social density. Overall, our studies shed light on how social stress regulates brain functions and behaviors.
Characterizing Short-Term Linear and Nonlinear Heart Rate Variability (HRV) in Duchenne Muscular Dystrophy
Alexs A. Matias* (University of Delaware), Krista M. Szymanski (University of Delaware), Michele N. D’Agata (University of Delaware), Mena Scavina (Nemours Children’s Hospital), Michael A. McCulloch (University of Virginia), David G. Edwards (University of Delaware), Melissa A. Witman (University of Delaware)

Duchenne muscular dystrophinopathy (DMD) is a rare muscle wasting disorder with a high mortality rate, predominately of cardiac origin. Heart rate variability (HRV) is a robust measure of cardiac autonomic balance known to be impaired in DMD and is typically reported in time and frequency domains. However, nonlinear HRV is associated with the structure of heart rate dynamics providing complementary diagnostic information on pathological HRV but these data are limited. PURPOSE: To characterize autonomic balance of the heart using both linear and nonlinear analyses of HRV in boys (ages 7-21) diagnosed with DMD as compared to typically-developing controls. METHODS: Error-free (<5 beats corrected) HRV during quiet rest was assessed in boys with DMD (n=6) and controls (Con) (n=16) using three-lead electrocardiograms for 5 continuous minutes. HRV was calculated from heart rate (HR) recorded on an individual beat basis and analyzed using linear (root mean square of the successive differences[RMSSD], high frequency[HF] and low frequency[LF] components) and nonlinear Poincáre plot analysis of SD1, SD2, as well as approximate entropy[ApEn]. RESULTS: HR was significantly higher in DMD (DMD: 95±9 bpm vs. Con: 67±13 bpm, p<0.001). RMSSD (DMD: 35±22 vs. Con: 85±43, p<0.001) and HF (DMD: 45±17 vs. Con: 63±16 %, p=0.013) were both reduced and LF/HF ratio was elevated in DMD (DMD: 1.22±0.50 vs. Con: 0.63±0.41, p=0.021). SD1 (DMD: 35±16 vs. Con: 60±31 ms, p<0.001) was lower in DMD and SD2/SD1 ratio was increased (DMD: 1.92±0.35 vs. Con: 1.39±0.31, p=0.010). ApEn was also elevated in DMD (DMD: 1.25±0.08 vs. Con: 1.12±0.11, p=0.011). DISCUSSION: Preliminary findings suggest impairments in vagal tone, and a shift towards sympathetic dominance, in DMD from both linear and nonlinear analyses. Future directions include investigating the changes in nonlinear HRV across time to determine if they can provide further insight into the cardiac disease progression in patients with DMD.
Enhancing Physical Rehabilitation: Optimized Balance Training Through Augmented and Virtual Reality
Gavin Caulfield* (University of Delaware)

As we expand the virtual and augmented reality world, we must ask, which would best suit as a strong alternative to balance training. Balance training is an essential part of physical therapy and rehabilitation as it can vastly improve a patient’s mobility and coordination. However, traditional methods disregard two essential components of successful rehabilitation: motivation and personalization. In this work, we present the optimized substitute for balance training through lower extremity stimulation within matching environments among both virtual and mixed reality. With a user study (N=8), we began each participant in a room with an augmented coach instructing the users with balance-specific poses to mimic, followed by another trial of the same nature but immersed in a virtually replicated room. The results revealed that the participants preferred augmented reality over virtual. Additionally, the data trended toward participants succeeding within augmented reality over virtual. Our work and results provide the direction of focus for the future of physical rehabilitation, along with an enjoyable alternative to the current “one-size-fits-all” that could fail patients who require lower-extremity rehabilitation–specifically stroke victims or sufferers of a gait abnormality.
Development of complete cell surfaceome for -omic analyses reveals potential immunotherapy targets for pediatric acute myeloid leukemia
Joshua R. Faust* (Nemours Childrens Health; University of Delaware), Anilkumar Gopalakrishnapillai (Nemours Childrens Health; University of Delaware), Sonali P. Barwe (Nemours Childrens Health; University of Delaware)

Multiple “omics” based approaches are currently utilized to gain deeper understanding about biological processes. Several platforms provide curated gene sets for gene ontology-based pathway mapping and functional annotation. Protein localization provides clues about its role. For example, cell surface proteins often function as receptors for signal transduction, mediate cell-cell or cell-matrix interactions and/or facilitate assembly of signaling complexes. Thus, changes in cell surfaceome comprising all proteins localized to the cell surface between experimental groups could shed light on biological mechanisms. Currently, a comprehensive surfaceome list is not available.
The Universal Protein Resource (UniProt) can pull out membrane-associated proteins, but it is not comprehensive, as it cannot recall some cluster of differentiation (CD) proteins and non-transmembrane proteins. Other methods utilizing machine learning approaches to predict the surfaceome missed several transmembrane proteins. We have developed a comprehensive cell surface proteome list by isolating transmembrane proteins from UniProt and combining them with a surfaceome list from literature through machine learning approaches. After combining source lists, genes were filtered to remove duplicates and mitochondrial membrane proteins, leaving roughly 5,400 protein coding genes implicated to have subcellular localization to the cell membrane.  We applied the list to differential gene expression analysis of pediatric acute myeloid leukemia (AML) from Therapeutically Applicable Research to Generate Effective Treatment (TARGET)-AML gene expression data. Surfaceome filtering of genes differentially expressed in pediatric AML compared to normal specimens revealed established immunotherapeutic targets, like MSLN, CD70 and UNC5C. Common AML targets, CD123 and CD33, have increased expression in AML bone marrow, but are also expressed in healthy bone marrow (not differentially expressed), so targeting these proteins results in significant toxicities. We and others have demonstrated preclinical efficacy of MSLN-targeting immunotherapy approaches in preclinical models, which are now in clinical trials. Thus, characterization of differentially expressed surfaceome genes will identify novel therapeutic targets.
Assessing the Effects of Dual Tasking on Force Output During Bimanual Coordination: A Pilot Study
Sophia Crisomia B.Sc.*, Abigail Bower M.A., Roxana Burciu Ph.D (University of Delaware)

Background: The ability to coordinate hand movements is crucial for a wide range of daily activities. Bimanual activities require individuals to exert different forces and are often performed under conditions where visual feedback related to the task may or may not be available, or while attending to another task. Despite extensive research examining bimanual coordination under various force and feedback conditions, we have a limited understanding of how individuals coordinate their hands while performing a concurrent, cognitive task. Therefore, the purpose of this study is to examine the effects of dual tasking on force output during a bimanual coordination task.
Methods: Four young adults (YA) (age: 23.75±1.71) performed a visually guided isometric bimanual coordination task that involved generating 10, 20, and 30% of their maximum voluntary contraction (MVC), with and without visual feedback. Furthermore, participants had to complete this task while also simultaneously engaging in a cognitive activity that required them to mentally count the number of letters they heard.
Results: Absolute error (AE) increased without visual feedback for both the dual- and single-tasks for each of the MVC levels. AE was increased at higher force levels and this effect was greater in the no feedback condition. During the feedback condition, AE was greater in the dual task than in the single task across all force levels. However, this was not the case for the no feedback condition.
Conclusions: YA are less accurate when generating higher forces and without visual feedback. Preliminary data shows that the accuracy of movement can decrease even further when simultaneously attending to a cognitive task. However, the interference from the cognitive task remains to be confirmed in a larger dataset. Finally, this dual-task paradigm will provide a baseline for comprehending how this process is altered in neurological patients who experience both motor and cognitive difficulties.
Impairment of endothelial Kir channels in response to elevated glucose is independent of changes in channel expression.
Masoumeh Sadeghinejad*, Erica Johnson, Ibra Fancher

Background and Objective. Obesity comprises a complex metabolic pathophysiology often associated with poor glycemic control. Our past studies detailed that impairment of inward rectifying potassium channels (Kir2.1) is a contributing factor to obesity-induced endothelial dysfunction, however, the major obesity-associated factors driving Kir2.1 impairment are unknown. As obesity is often associated with hyperglycemia, we aimed to determine the effect of elevated glucose on endothelial Kir2.1 channel function and expression in vitro.
Method. We treated human adipose microvascular endothelial cells (HAMECs) with high glucose (200mg/dL), mannitol (200 mg/dL), or NaCl (300 mOsm/kg) for 72 hours. Control cells were maintained in standard media containing 100 mg/dL glucose. Following treatments, Kir currents were measured using whole-cell patch clamp electrophysiology and Kir2.1 expression was assessed using qPCR, Western immunoblotting, and immunocytochemistry (ICC). ICC was performed on nonpermeabilized HAMECs to detect membrane expression. Statistics: A Kruskal-Wallis one-way analysis of variance (p < 0.05) followed by Dunn Sidak post hoc analysis (*) was used to determine significant differences.
Result. HAMECs exposed to elevated glucose exhibited reduced Kir current compared to control. Surprisingly, mannitol induced similar decreases in Kir current. As mannitol originally served as the osmotic control, we next tested the effect of osmotic stress on endothelial Kir current by exposing cells to elevated NaCl, which had no effect. Furthermore, increased glucose or mannitol did not influence channel KCNJ2 gene or total Kir2.1 protein expression, however, preliminary experiments indicate that the membrane expression of Kir2.1 may be increased following exposure to elevated sugars.
Discussion and Conclusions. Our findings indicate that glucose and mannitol reduce endothelial Kir currents without affecting channel expression, suggesting that functional impairment of the channels is induced by elevated sugars. This is further supported by preliminary studies that indicate that membrane Kir2.1 expression is increased in response to elevated sugars, suggesting that a possible compensatory trafficking mechanism fails to restore Kir2.1 channel function despite an increased membrane presence. As the observed decreases in Kir channel current are not mediated by an increase in osmotic pressure under these conditions, we aim to next test if damage to the glycocalyx, which is established to occur in obesity and renders endothelial Kir channels dysfunctional, occurs in the presence of elevated sugars.
Acknowledgement. This study was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under award number 2P20GM113125 (Ibra Fancher).
Identification of a heme acceptance domain in the System I bacterial cytochrome c biogenesis pathway
Alicia N Kreiman* (Univ. of Delaware), Molly C Sutherland (Univ. of Delaware)

Cytochromes c are highly conserved, critical electron transport proteins. In prokaryotes the various roles of cytochromes c contribute to the diversity of microbial metabolism and allow bacteria to survive and thrive in nearly every ecological niche. Proper protein folding and function of all cytochromes c requires a unique covalent attachment of heme to a conserved CXXCH motif in apocytochrome c. This process of heme attachment is known as cytochrome c biogenesis and all known cytochromes c are matured by one of three pathways: System I (prokaryotes), II (prokaryotes) or III (eukaryotes). Here, I focus on System I, which is composed of eight integral membrane proteins, CcmABCDEFGH, and is proposed to function in two steps. First, heme is trafficked from the unknown site of heme acceptance across the inner membrane by CcmABCD and attached to CcmE. Second, the ATP-hydrolysis driven release of holo-CcmE allows for the transfer of heme to CcmFH for attachment to apocytochrome c. In the absence of CcmAB, heme is still transported across the membrane and covalently attached to CcmE, but CcmE is not released. Thus, we hypothesize that CcmC is the heme receptor for the System I pathway. In this work I seek to identify a heme acceptance domain in the transmembrane protein, CcmC. This study leverages a well-developed recombinant E. coli expression system to perform cysteine/heme crosslinking throughout the putative heme acceptance region of CcmC. The objective of this technique is to ‘trap’ heme and map the CcmC cytoplasmic heme acceptance domain. To identify cysteine mutations that ‘trap’ heme, a biochemical approach utilizing affinity purification, heme staining, and UV-vis spectral analysis will be used. Preliminary results show that this approach is feasible in our system and these studies allow for further elucidation of the heme trafficking pathway through the bacterial inner membrane.
Associations between Peak VO2 and Sodium-induced Changes in Vascular Function
Serena A. Schade*, Jordan C. Patik, Justin D. Mehrer, Shannon L. Lennon, William B. Farquhar, David G. Edwards. University of Delaware, Newark, DE

Excess dietary sodium has been shown to reduce brachial artery flow-mediated dilation (FMD), a key indicator of conduit artery endothelial function. Although aerobic fitness is known to have vascular protective qualities, it is unknown whether increased aerobic fitness can mitigate the negative impact of excess sodium on the vasculature.
PURPOSE: To test the hypothesis that increased aerobic fitness is associated with less change in FMD and reactive hyperemia (RH) following short-term, sodium loading (SL).
METHODS: Twenty-three (13F/10M; Age: 26±4yrs.) healthy, young adults completed the study. Participants completed a graded exercise test on a cycle ergometer to determine peak oxygen consumption (VO2peak, range: 17.5 – 69.1ml/kg/min). Participants were instructed to continue their usual diet and were studied on two occasions, following 10 days of SL (10g of sodium chloride capsules) or placebo capsules, in random order. Brachial artery diameter and blood velocity were recorded via duplex ultrasound at baseline and following a 5-min occlusion of the forearm.
RESULTS: SL increased urinary sodium excretion (291.5±113mmol/24 hr) in comparison to the placebo (164.1±74.7 mmol/24 hr, p<0.05). FMD was unchanged across conditions (PL: 8.5±3.5%, SL: 8.7±3.0%, p=0.81) and no relation was observed between VO2peak and ∆FMD (r=0.03, p=0.90). However, SL reduced total RH (∆38±61 ml, p<0.01). VO2peak was inversely related to the change in peak RH (r=-0.53, p=0.01) and total RH (r=-0.57, p=0.007).
CONCLUSION: Contrary to our hypothesis, these preliminary data suggest that aerobic fitness does not mediate sodium-induced changes in conduit artery endothelial function when assessed by FMD. Our RH data, however, indicates that increased aerobic fitness is associated with greater consequences of excess sodium on microvascular function. Further research is needed to understand the interactions between aerobic fitness and excess sodium on vascular function.
Supported by NIH Grants 5R01HL104106 and 5P20GM113125; AHA 20POST35080171.
Hyperglycemia Degrades the Endothelial Glycocalyx Through a Decrease in Syndecan-1
Emma C Hudgins* (University of Delaware), Erica J Johnson (University of Delaware, Adam M Bonar (University of Delaware), Ibra S Fancher (University of Delaware)

Although endothelial dysfunction is known to be associated with obesity, the mechanisms causing the dysfunction are not well understood. The glycocalyx is an extracellular matrix present in endothelial cells that has been shown to play a role in endothelial cell function. It is composed of glycosaminoglycans anchored to membrane proteins such as Glypican-1, which is thought to play a role in mechanosensing, and Syndecan-1, which has been shown to be involved in physiological endothelial function. To assess the effect of hyperglycemia on the glycocalyx, cultured endothelial cells were treated with high levels of glucose. Gene expression was assessed with PCR, and protein expression was evaluated with Western blot and fluorescent immunostaining. No change was observed in either Glypican-1 or Syndecan-1 gene expression after exposure to high glucose. Western blot showed a decrease in whole-cell Syndecan-1 expression after glucose treatment, and immunocytochemistry data showed a similar decrease in membrane expression of Syndecan-1. No significant change was noted in whole-cell or membrane protein expression of Glypican-1. This may suggest that hyperglycemia contributes to endothelial dysfunction by degrading the glycocalyx through a decrease in Syndecan-1. Further experiment is needed to determine whether this decrease is due to a downregulation of translation or an increase in Syndecan-1 shedding.
Stress & the Developing Brain: An MRI Pilot Study
Taylor S. Campbell* (Univ of Delaware), Bernie Chen (Univ of Delaware), Mary Dozier (Univ of Delaware), & Tania L. Roth (Univ of Delaware)

Our experiences during early development can stay with us throughout our lifespans by affecting the way the brain develops over critical and sensitive periods. Animal models indicate that experiencing stress particularly affects brain development by interfering with neural processes, like synaptic pruning and myelination. In humans, these disruptions in early neural circuit programming may leave a child more susceptible to developing psychiatric disorders as they age. Magnetic resonance imaging (MRI) provides a unique opportunity to investigate how these early disruptions may be affecting the developing brain at the structural level (i.e. regional brain volume), as well as the microlevel (i.e. white matter integrity). We conducted two novel pilot studies to investigate 1) if disruptions in the infant-caregiver relationship affect regional brain volume and white matter integrity in adolescent and young adult female rats and 2) how aerobic exercise during adolescence may influence the developing brain in this same context. To do this, we utilized a validated rodent model of early life stress wherein one rodent litter was randomly assignment to a nurturing care condition where they were placed with a lactating foster dam who was provided with ample nesting and bedding material, or a lactating foster dam in an impoverished or resource-scarce environment. During young adolescence, these rat pups either remained under standard housing conditions or were given access to voluntary running wheels for 20 days. All MRI scanning took place during early adolescence and young adulthood to capture 1) adolescent-adult changes in brain development in the context of early life stress and 2) pre- and post- exercise-induced brain changes (within the exercise treatment groups). MRI data collection is completed, and data preprocessing and analyses are currently underway. MRI data collection for this project was supported by COBRE pilot grants awarded to the University of Delaware.
The Association Between Cardiovagal Baroreceptor Sensitivity and Hippocampal Tissue Integrity in Young and Middle-aged Adults
Fiona M. Horvat* (University of Delaware), Mary K. Kramer (University of Delaware), Curtis L. Johnson(University of Delaware), Megan M. Wenner (University of Delaware), Christopher R. Martens (University of Delaware)

Prior research has investigated the association of cardiovagal baroreceptor sensitivity (BRS) with white matter neuronal integrity and cerebral perfusion using magnetic resonance imaging (MRI); however, less is known about the association with specific regions of gray matter (i.e, hippocampus) involved in memory formation. MR elastography (MRE) has emerged as a constructive tool for assessing the viscoelastic mechanical properties of the brain, which reflect the microstructural integrity of neuronal tissue. Previous studies have yet to determine whether BRS and hippocampal (HC) stiffness are correlated. As a result, this study seeks to examine the association between cardiovagal BRS and viscoelastic properties of the brain, with the sub-goal of examining how advanced age affects this relationship. We hypothesized that there would be a positive relation between cardiovagal BRS and HC viscoelastic properties that strengthens with age, indicating a greater influence of blood pressure control on HC microstructural integrity. Cardiovagal BRS and HC viscoelastic properties were measured in 10 young (25 ± 2 years) and 10 middle-aged adults (55 ± 3 years). As expected, we observed lower cBRS in the older group than young (p≤0.05). There were no significant differences in HC stiffness or damping ratio when examined between age groups (p=0.69; p=0.56). However, a multiple linear regression with age included as a categorical covariate indicated that the middle-aged group tends to have a stronger relationship between HC properties and cBRS than the young group (p=0.07). In contrast to our hypothesis, preserved BRS was associated with lower HC stiffness in the middle-aged group; however, the physiological importance of this finding needs to be more completely explored. Our findings indicate that the association between short-term blood pressure regulation via cardiovagal BRS may be more closely linked to HC tissue integrity with advancing age. These mechanisms should be explored in a larger cohort including older individuals.
The Influence of Cholesterol on Resting Brain Blood Flow and Cognition
Kevin P. Decker* (University of Delaware), Faria Sanjana (University of Delaware), Nick Rizzi (University of Delaware), Mary K. Kramer (University of Delaware), Alexander M. Cerjanic (University of Delaware), Curtis L Johnson (University of Delaware), Christopher R. Martens (University of Delaware)

Objective: The purpose of this project was to determine the influence of cholesterol on brain blood flow and cognition.
Methods: Total, High-Density Lipoprotein (HDL), and Low-Density Lipoprotein (LDL) cholesterol was collected in 18 middle-aged adult participants (57 ± 4 years old). Resting brain blood flow was assessed using a pseudo-continuous arterial spin labeling (pCASL) sequence from the Human Connectome Project with a 64-channel head coil inside a Siemens Prisma 3T MRI scanner. Cognition was assessed using the Hopkins Verbal Learning Test (HVLT), Flanker Inhibitory Control and Attention Test, and Pattern Comparison Processing Speed Test.
Results: Total cholesterol (207 ± 28 mg/dL; r = -0.71, r2 = 0.51, p = 0.0009) and LDL cholesterol (123 ± 23 mg/dL; r = -0.78, r2 = 0.60, p = 0.0001), but not HDL cholesterol (67 ± 17 mg/dL; r = -0.03, r2 = 0.001, p = 0.9), were significantly associated with resting brain blood flow (70.0 ± 12.5 ml/min/100g). Among the cognitive testing, only memory retention from the HVLT was positively associated with HDL cholesterol (r = 0.51, r2 = 0.26, p = 0.03). A follow-up analysis revealed participants with a higher resting brain blood flow presented a tendency for higher scores in the Pattern Comparison Processing Speed Test (r = 0.44, r2 = 0.20, p = 0.06).
Conclusion: Two cardio-metabolic risk factors, elevated total and LDL cholesterol, were negatively associated with resting brain blood flow whereas the potentially cardioprotective HDL cholesterol was not. However, HDL-C was positively associated with the cognitive domain of word memory recall. Our results suggest cholesterol may impact resting brain blood flow and cognitive function, but additional data is required to elucidate the underlying mechanisms.
Grant Support: NIH/NIGMS – P20 GM113125
Investigating the active site of CcsBA, the bacterial holocytochrome c synthase
Tania Yeasmin*(University of Delaware), David J. Hawtof(University of Delaware), Molly C. Sutherland(University of Delaware)

Prokaryotic organisms possess the ability to survive in diverse, sometimes harsh conditions due to their genetically encoded unique electron transport chains and metabolic pathways. Cytochromes c are highly conserved proteins that function in these electron transport chains, responsible for critical cellular functions such as respiration, photosynthesis, and detoxification. Nearly all living organisms encode cytochromes c and the uniqueness of this protein resides in its requirement for a covalently attached heme at the conserved CXXCH motif of apocytochrome c (both of which needs to be in a reduced state to form the thioether bond) for proper folding and function, a process known as cytochrome c biogenesis. Three different pathways: System I (prokaryotes), System II (prokaryotes), and System III (eukaryotes) have been identified as accomplishing cytochrome c biogenesis. In this study, we focus on the System II pathway (composed of two proteins CcsB & CcsA) to determine if the key heme-handling residues of Helicobacter pylori and Campylobacter jejuni CcsBA are conserved with the well-characterized Helicobacter hepaticus CcsBA. Sequence alignment of CcsBA proteins from different organisms reveals that despite low sequence homology, the key heme handling residues are the same across CcsBA’s. Prior work has shown that CcsBA has two transmembrane histidines that act as an axial ligand to heme and two periplasmic histidines that act as an axial ligand to heme bound to well-conserved WWD domain. To determine if these residues work in a similar manner, site-directed mutagenesis was used to introduce glycine or cysteine at these key positions, followed by biochemical and functional assay to determine the functional conservation of the key heme handling residues, revealing the key features of heme handling across organisms.
The combination of violet light and infra-red as well as violet light only effectively suppress the survival of multiple-drug resistant E. coli.
Matthew Stangl* (University of Delaware), Dinesh K. Verma (Delaware State University) , Yong-Hwan Kim (Delaware State University)

Since the recent global pandemic started, there has been a high demand for establishing an inexpensive but effective method to interfere with the spread of infectious diseases. In accordance with our previous study, several combinations of 24-Watt violet light (V, 405 nm) with infra-red (IR, 850 nm) were used to identify an optimal light for suppressing multiple-drug resistant (MDR) E. coli at distances of 50 cm, 1 m, and 2 m. In this study specifically, we compared 24W violet and infra-red lights (used in the previous study) against 50W violet light (V, 405 and 410 nm) and infra-red (IR, 850 nm). Our results demonstrate that both 24W and 50W 4V and 3V-1IR (3:1 ratio in combination of violet and infra-red) effectively suppressed all MDR-E. coli growth tested. In addition, both 24W and 50W 4V and 3V-1IR nearly equally terminated MDR-E. coli effectively. Equal termination of MDR-E. coli colonies was also seen when comparing 50W 3V-1IR at wavelengths of 405 and 410 nm. Because of these results, we suggest using 50W 3V-1IR light (410 nm) for practical applications due to less safety concern. This is because violet light of wavelength 410 nm is further in the visible spectrum away from UV range and generally regarded as being safer for humans . In addition, although the same effectiveness in terminating MDR-E. coli colonies, 50W light provides more practicality as it creates a greater cone of illumination, allowing the light to cover a greater surface area and suppress a higher overall number of colonies. Notably, these results suggest that the combinations of violet and infra-red light can be used as a highly effective and inexpensive way to suppress the growth of MDR bacteria in a variety of settings, such as high contact surfaces in hospitals, nursing homes, and public spaces with high human foot traffic.
Cognitive Impairment but not Age affects Split-belt Locomotor Learning
Soumya Bhat, PT, DPT*, Matthew L Cohen, Ph.D., and Susanne M Morton, PT, PhD

Motor learning can be explicit or implicit. Explicit learning involves conscious effort and strategy (i.e., requires cognition), while implicit learning is subconscious and automatic. Recently, we have shown that cognitive impairment negatively affects some aspects of explicit motor learning, but it is not clear whether it affects implicit locomotor learning. Here, we used the split-belt treadmill locomotor learning paradigm, which is well-known to induce implicit learning, to compare locomotor learning among older adults with mild cognitive impairment (MCI; n=13), age-matched healthy older adults (OA; n=13), and healthy young adults (YA; n=12). We hypothesized that age, but not cognitive impairment, would negatively impact implicit split-belt locomotor learning. A single session of split-belt treadmill locomotion was conducted, during which participants walked with the belts split (moving at two different speeds) at a 2:1 speed ratio to induce asymmetric step lengths. Over the course of learning, individuals slowly adjust their walking pattern to restore step symmetry despite continued asymmetric leg speeds. The primary outcome was learning magnitude, or the mean normalized step length asymmetry at the end of learning. We compared learning magnitudes among the three groups and assessed correlations in the YA group and the pooled OA and MCI groups to determine if split-belt locomotor learning was related to cognition. We found that the MCI group demonstrated impaired locomotor learning compared to age-matched OA. Further, in OA and MCI groups, poor cognitive performance was related to worse implicit learning. We conclude that forms of motor learning thought to act predominantly implicitly may, in fact, rely more on top-down cortical processes, especially in OA and cognitively impaired groups. Clinicians should not assume that motor learning processes work similarly in young and older adults. Future research should investigate if these learning deficits predict cognitive decline in these populations.
The Effect of Dietary Sodium and Potassium Consumption on Intestinal Permeability, Blood Pressure, and Endothelial Function: A Pilot Study
Kathryn E. Kaseman* (University of Delaware), Andrea J. Lobene (University of Delaware), Michael R. Axler (University of Delaware), Kristina D. Krieger (University of Delaware), Shannon L. Lennon (University of Delaware)

High sodium (HS) diets can impair endothelial function (EF) and increase blood pressure (BP) while high potassium (HK) diets are shown to counter this effect. Furthermore, HS diets may lead to gut dysbiosis. Dysbiosis can increase intestinal permeability (IP), activating pro-inflammatory pathways, which can impair EF and increase BP. HS diets have been shown to cause dysbiosis, activate pro-inflammatory pathways, and increase BP in men. However, whether sex differences exist in response to HS-induced changes to IP are unknown as well as the role of HK diets. Twelve healthy participants (6W/6M, age 32±8 yrs, SBP 104±7/66±6 mmHg) consumed three 10-day diets of differing sodium and potassium quantities in a randomized order: low sodium (LS)/moderate potassium (MK) (LS/MK: 50mmol/65mmol), HS/MK (300mmol/65mmol), and HS/HK (300mmol/120mmol). On day 10, mean arterial BP (MAP) and brachial artery flow-mediated dilation (FMD), were measured. Blood was collected and lipopolysaccharide-binding protein (LBP), a biomarker of IP, was measured by ELISA. While LBP was increased on the HS diets, it did not reach statistical significance (diet: p=.08; LS/MK: 5.0±3.7 μg/ml; HS/MK: 10.0±6.8 μg/ml; HS/HK: 9.3±5.4 μg/ml) while men had increased LBP compared to women (sex: p<.01). FMD was not different between diets (p=.10) although men had a lower FMD compared to women after each diet (sex: p<.01). MAP was not different between diets (p=.21), and only men demonstrated changes to MAP between diets (diet*sex: p=.03). LBP was inversely related to FMD (r=-0.82 p<.01) and positively related to MAP (r=0.72 p<.01) on the HS/MK diet for all subjects, but not the LS/MK or HS/HK diets. These pilot data suggest that, in the context of a HS/MK diet, greater IP is associated with lower FMD and higher BP in all subjects and these relations were primarily driven by men.
Azacitidine chemosensitizes KMT2A rearranged pediatric AML by modulation of cell surface proteins to promote mobilization from the bone marrow
Kara Lehner* (UD, Nemours) , Josh Faust (Nemours), Anilkumar Gopalakrishnapillai (Nemours) and Sonali Barwe (UD, Nemours)

KMT2A is the most common chromosomal translocation in pediatric acute myeloid leukemia with a poor prognosis. The survival rate is less than 50% due to frequent relapse and limited response to chemotherapy. We identified interactions between cells in the bone marrow microenvironment help protect KMT2A rearranged AML cells from chemotherapy and contribute to leukemia progression. We also showed that epigenetic drug combination azacitidine and panobinostat chemosensitizes KMT2A rearranged AML cells by decreasing adhesion to endothelial, stromal and mesenchymal stem cells and initiating AML cell mobilization to peripheral blood in vivo.
To characterize the mechanism of chemosensitization by the epigenetic drug combination, we analyzed the modulation of gene expression signature by azacitidine and cytarabine in KMT2A rearranged PDX cells treated in vivo. We also identified cell surface proteins that were differentially expressed following cytarabine and azacitidine treatment. This analysis showed that following azacitidine treatment expression of a cell surface protein directly upregulated by KMT2A fusion protein, SCUBE1, is downregulated resulting in leukemic cell death. Additionally, we identified reduced expression of integrins, chemokine receptors and cadherins including CXCR7, ADAM23 and cadherin 24 following azacitidine treatment. In summary, our data shows azacitidine modulates cell surface proteins to initiate mobilization and chemosensitization in KMT2A rearranged AML.
Characterization of NAD-decapping enzyme: NudC
Yogeshwari Singh* (Univ of Delaware), Henry Anderson (Univ of Delaware), Lingting Li (Univ of Chinese Academy of Sciences) Imalka Mudiyanselage (Wayne State Univ), Jared Schrader (Wayne State Univ), Yu Zhang (Univ of Chinese Academy of Sciences), Karl Schmitz (Univ of Delaware), Jeremy G. Bird (Univ of Delaware)

RNA stability is strongly influenced by the identity of its 5′ ends. Besides the canonical 5′ methyl-7-Guanosine (m7G) cap in eukaryotes, various primary metabolites that can be added to the 5′ end of RNA including Nicotinamide Adenine Dinucleotide (NAD+/NADH), dpCoA, FAD, dinucleotide polyphosphates, and UDP-GlcNAc, and UDP-GlcNAc. These metabolites are used by both prokaryotic and eukaryotic RNA polymerases to initiate transcription as non-canonical initiating nucleotides (NCINs). In E. coli, NAD added at the 5′ end of RNA transcripts results in a longer half-life of RNA by protecting them against degradation from nuclease enzymes in the cell. Members of the Nudix hydrolase family, like E. coli NudC, Saccharomyces cerevisiae NPY1, and eukaryotic Nudt, remove these caps and leave a monophosphate 5′ end, which subjects the RNA to be degraded by RNases such as RNAse E in E. coli. The addition and removal of NCIN caps by RNAP and by NudC, respectively, suggest a regulated process affecting the cell’s transcriptome under different conditions. Here, I will present the mechanism of decapping and characterization of the NudC enzyme by:
1. Biochemical and structural data characterizing the substrate specificity of E. coli NudC showing NudC has different specificity towards different substrates that can result in the regulation of various RNA. Crystal structures of NudC- apo and in complex with different caps show the active site and interaction with the caps.
2. Evidence that NudC decapping activity is regulated by the alarmone ppGpp. An increase in ppGpp upregulates NudC in cells during the stress response.
3. In-vivo knockout evidence shows that NudC and the closely related enzyme RppH act as primary decapping enzymes in E. coli.
Understanding this balance between RNA stability and degradation during growth and stress phases is important to shed light on the functions of non-canonical capping and its relation to metabolism.
TMS for Response Inhibition in Pediatric OCD: Methods and Rationale
Mitchell Jackson*, B.S.1 (Bradley Hospital), Nicole McLaughlin, Ph.D.2,3 (Butler Hospital), Eric Tirrell, B.S.3 (Butler Hospital), Zoe Brown, B.S.1 (Bradley Hospital), Linda Carpenter, M.D.2,3 (Butler Hospital), Kristen Benito (Bradley Hospital), Ph.D.1,2

Impaired response inhibition (RI; ability to suppress irrelevant or interfering behaviors) is a core deficit in pediatric OCD thought to contribute to the occurrence of compulsions. Strategies that can be used to improve RI offer a promising approach for augmenting response to existing OCD treatments. Neural connectivity between networks nodes involved in RI is well-understood (rIFG, right inferior frontal gyrus; presupplementary motor area, pSMA; subthalamic nucleus, STN). Transcranial magnetic stimulation (TMS) of pSMA, a key cortical node in the RI circuit, can acutely alter RI behavior and activity in functionally connected areas. Electroencephalogram (EEG) correlates with RI behavior and circuitry following TMS; a fronto-central N2/P3 signature consistently relates to RI task performance in adults and youth, and newer work suggests that beta band changes during these tasks can be used to index RI network connectivity. However, studies have yet to test whether pSMA stimulation with TMS can produce acute changes in the RI network and/or RI behavior in youth–and to our knowledge, there are no TMS treatment studies in youth with OCD at any neural target. We will present the methods and rationale for an ongoing pilot study aiming to begin addressing these gaps. Design. Within-subject, counterbalanced crossover design comparing TMS vs Sham in a brief 2-visit protocol enrolling 14 youth (age 13-18) with OCD. At each visit, youth complete the Stop Signal Task (SST) with concurrent EEG pre- and post- TMS or Sham. TMS is delivered over pSMA using continuous TBS (cTBS). After each visit, youth rate symptoms using ecological momentary assessment (EMA). Primary outcomes of interest include safety and tolerability, changes in EEG during SST stop trials (frontocentral P3 amplitude, beta band power in right frontal electrodes, phase coherence between pSMA and rIFG), changes in response time on SST stop trials, and changes in clinical symptoms.
Modeling gun violence and trajectory of behavioral health outcomes
Natalie Cartwright* (Norwich University), Nicole Cook (OCHIN), Alan Cook (University of Texas Health Science Center at Tyler), Kerime Toksu (University of Vermont), Fran Biel (OCHIN), Megan Hoopes (OCHIN), Ali Al Bataineh (Norwich University), David Hosmer (University of Massachusetts at Amherst), Turner Osler (University of Vermont), Michelle Gordon (OCHIN)

Injury from firearm violence is a growing public health and racial justice problem in the US, yet our understanding of firearm violence on health is incomplete. Current understanding of firearm injury is largely limited to people who sustain acute care injury and present for care to emergency rooms and inpatient hospitals (ED/IP). This is because standardized coding for firearm injury (ICD 9/10) are limited to bodily trauma, impacting researchers ability to conduct research on people with health impacts that do not require acute care.  There is evidence that suggests exposure to firearm violence, with or without acute care injury, can lead to a host of physical and psychological sequelae. However, due to lack of data, the mechanisms of this are poorly understood. There is a critical need to more fully frame the public health problem of firearm injury and capture the longitudinal physical and behavioral sequelae of firearm violence. Primary care electronic health records (EHRs) are a rich source of longitudinal physical and behavioral health data that, when combined with AI/ML, has the potential to powerfully expand our knowledge of firearm injury.
Caregiver satisfaction with new service delivery formats of various ASD services over the course of the
COVID-19 pandemic: A SPARK dataset analysis
Jung-mei Tsai* (University of Delaware), Alydia Meinecke(University of Delaware), & Anjana Bhat (University of Delaware)

Children with ASD receive a multitude of educational, medical, and therapeutic services. At the onset of the COVID-19 pandemic, all ASD services came to a complete halt following strict lockdowns (Bhat, 2021). Services resumed in various delivery formats – remote, in-person, or hybrid over the first year. Caregivers of children with ASD from the SPARK study (N=6,393; Ages: 19 months – 18 years) completed an online survey on the impact of the pandemic on access to services and their satisfaction with service delivery (remote, in-person or hybrid) for various services (special education, speech, occupational/physical therapies, ABA, mental health, and medical). ASD services had not recovered fully to an in-person format in the first year. Certain service types, (e.g., Special education, ABA, Speech therapy, and PT/OT had recovered to in-person formats to a greater extent (% recovery=24-46.3%) compared to mental health or medical services (% recovery= 15-20%). This was also associated with an increase in caregiver satisfaction with services (SLT, PT/OT % improvement in satisfaction = 7.1-7.3%) showing slightly greater improvements compared to mental health and medical services (MH and MED % improvement in satisfaction = 4.6-5.7%). The relationship between caregiver satisfaction and service delivery format seemed to evolve in that newer formats were more accepted over time. Nevertheless, in-person format of service delivery was consistently associated with greater satisfaction followed by hybrid followed by online formats of service delivery. Caregiver satisfaction with services also varied as a function of service type/format. In conclusion, healthcare services for children with ASD were evolving rapidly during the course of the pandemic and was associated with caregiver satisfaction with services received. These findings have important implications for how service delivery formats need to be suited to the type of services offered and directly influence parent’s overall satisfaction about service quality.
Delaware Comprehensive Sickle Cell COBRE: Clinical and Data Informatics Core
Robin Miller MD* (Nemours Children’s Health, DE), Jeff Meyers MS (Nemours Children’s Health, DE), Erin Coyne MSN, CRNP (Nemours Children’s Health, DE), David Brousseau MD, MS (Nemours Children’s Health, DE)

There is a tremendous need to improve clinical and research infrastructure for patients with sickle cell disease (SCD). As a disease of minorities, there is a longstanding history of disparate healthcare utilization, delivery, and availability as well as access to and participation in clinical research. The Clinical Research and Data Informatics Core (CRDIC) builds on the strong clinical and informatics research infrastructure established by the COBRE-supported Delaware Comprehensive Sickle Cell Research Center (DE SCD COBRE). The CRDIC is a centralized platform that provides the resources required to support SCD research and clinical quality improvement efforts, including guidance and mentorship of junior investigators and expertise in data analytics and data science techniques. This integrated clinical and informatics core significantly improves the ability to efficiently conduct successful, high-impact SCD research throughout DE, protect human subjects, and ensure delivery of the highest-quality clinical care. Use of a SCD-specific common data model and a scalable EHR-informed Learning Healthcare System (LHS) supports clinical care and facilitates cohort discovery and data reporting for research. The primary aims for this core include: 1) Creation of a robust and efficient platform supportive of investigator-initiated SCD clinical and translational research through the development of project management team specific for supporting the conduct of SCD research in the state of DE and a clinical center of expertise to oversee SCD research in DE, ensure the protection of human subjects, and provide guidance and mentoring for junior investigators. 2) Creation of a quality/outcomes team paired with community-level support, charged with ensuring the delivery of cutting-edge, high-quality, evidence-based, safe clinical care to patients with SCD throughout Nemours and across DE. 3) Maintain a scalable EHR-informed SCD LHS for managing SCD clinical and research data while improving patient care and translational research efforts across a broad range of projects.
The Center for Rural Health Care Delivery Science
Sandra L. Wong* (Dartmouth Health, Geisel School of Medicine at Dartmouth), Cassandra H. Brugger (Dartmouth Health), Timothy E. Burdick (Dartmouth Health, Geisel School of Medicine at Dartmouth), Martha L. Bruce (Dartmouth Health, Geisel School of Medicine at Dartmouth), Allison J. Hawke (Dartmouth Health), Sally A. Kraft (Dartmouth Health, Geisel School of Medicine at Dartmouth), A. James O’Malley (Geisel School of Medicine at Dartmouth), Mark A. Creager (Dartmouth Health, Geisel School of Medicine at Dartmouth)

This new Centers of Biomedical Research Excellence award establishes the Center for Rural Health Care Delivery Science at Dartmouth Health (DH), one of the most rural academic medical centers in the United States. Nearly 20% of Americans live in non-metropolitan areas and experience poorer health outcomes, but research on rural health care has been limited by challenges associated with relatively small and geographically dispersed populations. A directed focus on the study of rural health care aligns with DH’s strategy to deliver equitable care to rural patients via innovative care models. Accordingly, our Center concentrates on understanding and solving challenges associated with the provision of health care in rural areas, focusing on a community engaged approach to research and allowing our investigators to translate their findings back to rural communities and inform policymakers about critical issues that need to be addressed.
Our overarching goals are to establish a multidisciplinary research program in rural health care delivery science and to provide expert mentorship and infrastructure to train and support a critical mass of early-career clinician-scientists as they transition into independent researchers. To achieve this goal, we have four specific aims:
(1) Integrate research expertise, mentorship, and community engagement to establish a Center for Rural Health Care Delivery Science;
(2) Enhance the transition of early career clinician-scientists into independent researchers through strong research support and dedicated mentoring;
(3) Assure accountability and achievement of milestones through the implementation of a rigorous program evaluation plan; and
(4) Leverage shared resources to support the Center for Rural Health Care Delivery Science as a self-sustaining scientific community.
The Center’s work and Research Project Leaders are supported by three Scientific Cores—the Statistics, Informatics, and Qualitative Methods Core, the Community Engagement and Outreach Core, and the Administrative and Mentoring Core.
The RI-INBRE Molecular Informatics Core
Chris Hemme* (University of Rhode Island), Janet Atoyan (University of Rhode Island), Bongsup Cho (University of Rhode Island)

The Molecular Informatics Core (MIC, formerly the Bioinformatics Core) is the data science core (DSC) for RI-INBRE with focus areas in DNA sequencing, bioinformatics and data science, molecular modeling, and 3D science visualization. The MIC complements the CRCF to provide a unified data generation to analysis pipeline for many biomedical processes. In addition, the MIC works with researchers to meet general data management needs such as access to high performance or cloud computing resources, software resources, data sharing and management strategies, and public access compliance. Based on researcher demand, NIH data science mandates, and trends in bioinformatics research, the MIC will continue to promote cutting edge technologies such as single cell and spatial omics, machine learning, and cloud computing.
Rhode Island IDeA Network of Biomedical Research Excellence Centralized Research Core Facility at the University Rhode Island IDeA Network of Biomedical Research Excellence Centralized Research Core Facility at the University of Rhode Island
Ang Cai* (Univ of Rhode Island), Janet Atoyan (Univ of Rhode Island)

The RI-INBRE Centralized Research Core Facility (CRCF) is supported by Rhode Island INBRE and participating institutions including University of Rhode Island (URI), Brown University, Bryant University, Providence College, Rhode Island College, Roger Williams University, Community College of Rhode Island, Johnson & Wales University, and Salve Regina University.
The RI-INBRE Centralized Research Core Facility (CRCF) is supported by Rhode Island INBRE and participating institutions including University of Rhode Island (URI), Brown University, Bryant University, Providence College, Rhode Island College, Roger Williams University, Community College of Rhode Island, Johnson & Wales University, and Salve Regina University.
Imaging Facility at Delaware State University
Michael Moore* (Delaware State University), Hacene Boukari (Delaware State University)

Optical Center for Applied Research, Department of Physics and Engineering, Delaware State University, Dover, DE 19901
The Imaging Facility at Delaware State University is a multi- user core facility that offers advanced imaging and spectroscopy instruments with scientific and technical expertise in optical microscopy, spectroscopy and computational image analysis. The mission is to advance transformative excellence in research, innovation and education and to provide a vital support structure to the faculty and students in specialized facilities.
The core facility houses state of the art optical and electron imaging, material characterization and spectroscopy instrumentation. The facility currently has three confocal microscopes; a Zeiss 780 point scanning confocal, a Crest Optics V3 spinning disk confocal, and a Horiba XPloRA Raman confocal microscope. The facility has a Wide-Field inverted Fluorescence and DIC microscope and several advanced bright field microscopes with extended contrast techniques. In addition, we have a Bruker Innova AFM, a Bruker Analysis NanoIR2-S AFM, and an FEI Quanta FEG 250 with EDS, and ESEM. The imaging facility also hosts a UV-VIS spectrophotometer, an ISS-K2 spectro-fluorimeter. Furthermore, the data collected is archived on a server housed with-in the facility and the data is analyzed on our power workstations in our Image Analysis room. We have several commercial software packages.
The facility hosts workshops for graduate and undergraduate students, including middle school, and high school field trips. Due to a new DSU/Agilent partnership we plan to increase our microscopy and spectroscopy footprint. We will be adding LD-IR spectroscopy for micro plastics analysis, FTIR, Raman bulk spectroscopy, flow cytometry and a high content screen confocal imaging platform. Finally, to fulfill our mission to educate students we plan to integrate a microscope laboratory course into the facility and offer more middle and high school field trips and workshops.
Delaware Center for Musculoskeletal Research – Multiscale Assessments Research Core
Mary Boggs*, Christine Stinger, Charles Riley, Faysal Haque, Liyun Wang (all University of Delaware)

The Delaware Center for Musculoskeletal Research – Multiscale Assessments Research Core is a newly established multi-user facility that houses state-of-the-art equipment to apply and assess physical cues and biological signals in musculoskeletal and other systems. The DCMR Research Core provides services, expertise and training in the areas of multiscale biomechanical loading and measurement, preclinical models, multimodal preclinical imaging and histology. The specialized staff are experts in the various offerings of the Research Core. While the Research Core specializes in musculoskeletal systems, its resources can address the broad needs of life science researchers at the University of Delaware and other academic institutions, non-profit organizations and the industrial research community.
Mitochondrial Function and In Vivo Imaging (MF-II) Core Facilitates Basic and Translational Cardiopulmonary Vascular Biology Research in Rhode Island
Peng Zhang* (Ocean State Research Institute), Junfei Wei (Ocean State Research Institute), Amy Princiotto (Ocean State Research Institute), Elizabeth O. Harrington (Ocean State Research Institute), Gaurav Choudhary (Ocean State Research Institute)

The vision of the CardioPulmonary Vascular Biology (CPVB) COBRE program is to develop effective approaches to prevent and treat vascular diseases affecting the pulmonary and cardiovascular systems through better understanding of disease mechanisms. The Mitochondrial Function and In Vivo Imaging (MF-II) Core of the CPVB COBRE was established to meet the increasing demands from the CPVB research community in Rhode Island. Understanding the mechanisms related to mitochondrial dysfunction in settings of heart and lung injury may lead to unique approaches to interrupt disease progression and enhance cardiopulmonary vascular repair. High-resolution in vivo imaging technologies such as echocardiography, laser doppler imager, micro computed tomography, and positron emission tomography scan have become indispensable tools in modern translational research that provide unique opportunities for real-time study of functional and biological processes in animal models non-invasively and longitudinally. The MF-II Core is the first scientific core in Rhode Island to offer these cutting-edge services with centralized state-of-the-art equipment. To date, the MF-II Core has provided excellent services to investigators in Rhode Island and the other IDeA states and enhanced their productivity and research impact. The MF-II Core also showed a steady growth of the deliverables and worth of services over the years and has an excellent research pool for core usages. As the program continues, the MF-II Core will continue to provide critical support to investigators to facilitate basic and translational cardiopulmonary vascular biology research excellence in Rhode Island and the other IDeA states.
University of Delaware Synthetic Chemistry Core Facility
Yinzhi Fang* (University of Delaware), Joseph M. Fox (University of Delaware)

The Synthetic Chemistry Core Facility enables multidisciplinary research on campus by designing and carry out custom synthesis projects. This service center helps to plan, organize, direct, and execute the synthesis of small molecule targets for University research programs. We provides publication quality experimental descriptions of synthetic procedures and publication quality characterization data, including 1H and 13 C NMR spectra, MS and HRMS data, and upon request FT-IR, HPLC, UV-Vis and other spectroscopic data. The service center collaborates with researchers to design and develop research protocols to assist students, professionals and faculty in carrying out synthetic experiments in their own laboratories. The fee for service work is at competitive price for both academic and industrial users.
The COBRE Center of Molecular Epidemiology Biorepository Core at Dartmouth: Comprehensive support for diverse and large-scale research.
Palys, TJ*, Christensen, BC, Passarelli, MN, Karagas, MR. Department of Epidemiology, Geisel School of Medicine Dartmouth, Lebanon, NH.

The COBRE Center for Molecular Epidemiology Biorepository Core at the Geisel School of Medicine at Dartmouth utilizes a combination of manual and robotic specimen processes and automated archival storage systems to provide support to members of the Dartmouth community and beyond. The Core performs an array of services including specimen fractionation (e.g., of blood and breastmilk), processing of a wide range of biologic samples (e.g., urine, stool, nail, teeth, hair, saliva), human and microbial nucleic acid extraction, tissue separation and fixation, as well as post-archival processing and referral management services. Since its inception in 2013, the Core has handled >400,000 aliquots, created about 300,000 aliquots and maintained a highly secure, annotated archive of >600,000 diverse specimens. The Biorepository Core capabilities are well positioned to fully support large and diverse studies which integrate genetic, epigenetic, metabolomic, metagenomic, metallomic, and a broad range of other analyses (Table 1). The Core has provided comprehensive support for large environmental health and cancer-related studies such as the New Hampshire Birth Cohort Study, New England Bladder Cancer Study, the New Hampshire Health Study, the Vitamin D and Calcium Polyp Prevention Study and the Environmental influences on Child Health Outcomes Program.
Bridging Synapses: Connecting Scientific Research and Clinical Practice in Patients Receiving Transcranial Magnetic Stimulation for Major Depressive Disorder
Eric Tirrell* and Linda Carpenter, MD (both authors are affiliated with each below)
Butler Hospital TMS Clinic and COBRE for Neuromodulation Research, Providence, RI, USA;
Warren Alpert Medical School of Brown University, Providence, RI, USA

Background: Recruitment for research studies from patients flowing through a clinical treatment service such as Transcranial Magnetic Stimulation (TMS) provides the ideal opportunity for biomarker investigations that help inform TMS practice. Since 2015, Butler Hospital researchers have been actively engaging patients undergoing TMS treatment for Major Depressive Disorder (MDD), inviting them to participate in research that don’t alter their course of naturalistic TMS care.
Methods: Consecutively evaluated/treated patients were offered participation in one or more clinical research studies that involved procedures prior to, during, and/or after their course of insurance-covered TMS Therapy. Research included brain imaging (MRI, MRS), collection of blood specimens, recording of physiological data (EEG, heart rate variability, blood pressure), computerized cognitive assessments, app-based ecological momentary assessment (EMA) of symptoms over time, or completion of other self-report surveys. Serial assessment of symptom severity via scales were done as part of clinical routine and scores were recorded in a registry-style database to facilitate merging of treatment outcomes (response, remission) with other research outcome variables.
Results: Consent and baseline research procedures were completed pending insurance authorization prior to starting TMS. Participation in research was generally well received and 216 TMS patients engaged in research since 2015 while many elected to participate in multiple studies. To date, 19 papers were published from the data generated by this arrangement.
Conclusions: Using this integrated approach to support clinical research can help inform the field by gathering naturalistic treatment data that reflects real-life patients and clinical practice. A registry database characterizing TMS patients and their outcomes is ideal for 1) evaluating durability of response and likelihood of retreatment; 2) addressing clinical questions that have not been the subject of systematic prospective study (e.g., concurrent medications, parameter selection, targeting methods); and 3) merging clinical outcomes with imaging or other research biomarker data obtained pre and post-TMS. Registry data mining can provide valuable insights about personalization of care and generate hypotheses for testing in prospective studies.
Cell Isolation and Organ Function (CIOF) Core Enhances CardioPulmonary Vascular Biology Research in Rhode Island
Elizabeth O. Harrington (OSRI/ PVAMC)*, Amy Princiotto (OSRI/ PVAMC), Peng Zhang (OSRI/ PVAMC), Valeria Zaratte (OSRI/ PVAMC), Gaurav Choudhary (OSRI/ PVAMC)

The Cell Isolation/ Organ Function Core provides a unique skill set and expertise to Rhode Island vascular biologists by providing quality assurance in isolation, characterization, and propagation of vascular derived cells and fibroblasts and cardiopulmonary organ function analyses. The centralization of the cell isolation and organ function measurements has helped investigators to minimize the variability thus provided uniformity in data acquisition throughout all COBRE Projects. Isolation, characterization, and propagation of the cells, morphometric analyses of cells and tissue, and organ function measurements are time-consuming and costly; thus the services provided by this Core have permitted the Investigators to focus their efforts on aspects of their research endeavors related to experimental design, execution and interpretation.
Cell Science Core
Lisa Glazewski*(Nemours Children’s Health), Sonali Barwe, PhD (Nemours Children’s Health)

The Cell Science Core (CSC) specializes in flow cytometry, cell sorting, cell culture, protein analysis, and small molecule analysis. Contact us to access our services and instrumentation.
Flow cytometry rapidly analyzes samples on a cell-by-cell basis and collects multiple parameters at once, allowing researchers to analyze thousands of cells in seconds. Common applications include immunophenotyping, cell activation, differentiation, gene expression, transfection efficiency, particle internalization, cell quantification, cell cycle analysis, as well as proliferation, apoptosis, and cell death assays. Our two flow cytometers are the Novocyte 3000 with 3 lasers and 13 fluorescent parameters and the Novocyte Quanteon with 4 lasers and 25 fluorescent parameters.
Our BD FACS AriaIII cell sorter is capable of sorting live or fixed cells. Its 3 lasers and 11 parameters allow researchers to identify, select, and sort cells of interest for a variety of downstream analysis or culture and expansion. Our staff operate the FACS instrument so the researcher can focus on their samples and results. Please reach out to us in advance to schedule a project consult prior to sorting.
This year we added the Luminex xMAP Intelliflex (Luminex) System. Luminex assays multiplex and analyze dozens of targets in a single well, saving costs, time, and precious samples. These targets can be proteins like cytokines, chemokines, growth factors, etc or Nucleic Acids like mRNA gene expression. CSC will offer these assays as a service, contact us for more information.
Our other services/instrumentation include project consultations and troubleshooting, a fully functional BSL-2 cell & tissue culture facility, the Zeiss Axio Observer fluorescence microscope, LI-COR Odyssey XF Imager and C-Digit Scanner, and Perkin Elmer Victor Nivo 5F Plater Reader. The CSC works closely with investigators to help design, perform, and analyze experiments.
Eric Hall (Nemours Children’s Health), Suzanne McCahan* (Nemours Children’s Health), H. Timothy Bunnell (Nemours Children’s Health), Chris Pennington (Nemours Children’s Health), Dan Eckrich (Nemours Children’s Health), Cara Reedy (Nemours Children’s Health), Eileen Antico (Nemours Children’s Health), Michael Peck (Nemours Children’s Health), Steve Vijayan (Nemours Children’s Health), Michael Schoenbeck (Nemours Children’s Health), Lynn Allen (Nemours Children’s Health), Brad Samuel (Nemours Children’s Health), McKenzie Camacho (Nemours Children’s Health), Evan Miller (Nemours Children’s Health)

The Biomedical Research Informatics Center (BRIC) offers researchers specialized computing resources, support, training, biostatistics, data analysis, and data management. The team has extensive expertise in data analysis and machine learning applications. With the support of an NIH RECOVER grant, the team is developing natural language processing methods for extracting information from EHR notes. BRIC provides support for electronic health record (EHR), PEDSnet and National COVID Cohort Collaborative (N3C) data requests and analysis. The team maintains and manages REDCap, a secure web application for building and managing surveys and databases. BRIC is supported by Nemours and external grants such as DE-CTR ACCEL, DE-INBRE, PEDSnet, Sickle Cell COBRE, REACH COBRE, PRESERVE and others.
Implementing a Data Science Core for the Center of Integrated Biomedical and Bioengineering Research
W. Kelley Thomas* (University of New Hampshire), Anthony Westbrook (University of New Hampshire), Qi Zhang (University of New Hampshire), Krystalynne Morris(University of New Hampshire), Joseph Sevigny (University of New Hampshire), Lawrence Gordon (University of New Hampshire), Rick H. Cote (University of New Hampshire)

Advancing multidisciplinary biomedical research is significantly enhanced when there is support for rigorous experimental design, analysis, and management of the increasingly vast datasets generated.
During Phase 1 of the Center of Integrated Biomedical and Bioengineering Research, UNH invested in significant computational resources in the Research Computing Center (RCC) and together with the Hubbard Center for Genome Studies (HCGS) established effective support for bioinformatic needs. However, important gaps persist in biostatistical support services (including experimental design and implementation) as well in the management and sharing of large multi-omics datasets.
The objective for the Data Science Core (DSC) is to enable researchers to adopt multi-omics technologies that enhance opportunities for rigorous, multi-disciplinary research programs by: (1) providing comprehensive support infrastructure for data analytics throughout the entire data life cycle to address the rapidly increasing need for statistical analysis and management of data generated by multi-omics technologies and to promote the FAIR principles of data science by implementing best practices for data management and deposition; (2) creating training opportunities needed to support best practices in data science; and (3) implementing a business plan to ensure the long-term sustainability of the DSC infrastructure at UNH.
Establishment of a DSC that combines a full range of biostatistical and bioinformatics support services, management of computational resources (including support for cloud computing), and provision of training workshops will have a major positive impact for CIBBR investigators, the UNH research community, and our IDeA-state collaborators. This will be achieved by supporting innovative data analytics approaches necessary for multidisciplinary, multi-omics research, by enhancing rigor and reproducibility, and by combining strong institutional support with cost-recovery measures to become a sustainable permanent core facility enhancing biomedical and bioengineering research capacity at UNH.
BERD: A Biostatistics Epidemiology Research Design core for the Delaware IDeA Network
Claudine Jurkovitz* (ChristianaCare Health Services, Inc.; Delaware ACCEL-CTR; Delaware INBRE)

The Biostatistics, Epidemiology Research Design (BERD) core of the Delaware ACCEL Center for Translational Research (CTR) was created in 2013 and has matured into a large, stable yet dynamic core with a repertoire of skills and resources that enables quality clinical and translational (C&T) research. The BERD core includes biostatisticians, epidemiologists, data scientists, and bioinformaticians with a wide range of expertise from all four ACCEL partner institutions: University of Delaware, Nemours Children’ Health system, ChristianaCare Health Services Inc., and Delaware State University. Through the BERD, investigators find collaborators to assist on research design, data extraction and statistical analysis, explore cohorts and train on approaches ranging from basic statistics to cloud-based large scale genomic analyses. To expand its reach to all IDeA network researchers, BERD also serves as an INBRE affiliated core and is a resource for several COBREs.
In the next five years, BERD scientists will focus on several objectives: 1) Expand the consultative services for C&T and population health sciences. BERD will leverage the expertise in machine learning, natural language processing, cloud computing, and geospatial analysis that already exists in Delaware Institutions to make this expertise available to all Investigators; 2) Promote virtual self-paced synchronous and asynchronous learning opportunities. We will keep offering scientific lectures and will vet on-line material provided by other CTRs for posting on our Education hub and 3) Expand the biomedical informatics research infrastructure by leveraging expertise and skills across Delaware institutions to establish an integrated network of resources. We will develop teams that include clinicians with content knowledge and computer scientists with expertise in Artificial Intelligence and in working in the cloud environment.
In conclusion, BERD assembles a diverse core of skilled biostatisticians, epidemiologists, and bioinformaticians from each Delaware Institution who have the common goal of advancing translational science and assist C&T investigators.
The Pediatric Research Optimizing Methods in Stakeholder Engagement (PROMISE) Research Core: Institutional supports for community-engaged research.
J. J. Cutuli, PhD (Nemours Children’s Health), D. A. Collins, BS* (Nemours Children’s Health), J.Jung, BS (Nemours Children’s Health), A.E. Kazak PhD, ABPP (Nemours Children’s Health),
M.A. Alderfer PhD (Nemours Children’s Health)

The Pediatric Research Optimizing Methods In Stakeholder Engagement (PROMISE) Core supports the Research Expanding Access to Child Health (REACH) COBRE by facilitating investigations that engage communities and investigators as equals. Communities are powerful and essential partners in the effective, comprehensive, and equitable transformation of pediatric care. Addressing persistent income- and race-related disparities in child health and wellbeing requires an appreciation of factors across the child’s social ecology such as family-level resources, beliefs, trust, provider and health care system approaches, systemic disadvantages, and public policies. Communities carry forward an accumulation of the effects of and responses to these factors which have implications for health, health care, and research participation. Despite its power, many investigators cannot execute community-engaged research because it requires advanced frameworks and tools rarely covered in training nor supported by institutional infrastructure. Trainees also often lack established connections to communities. Without support, many junior investigators are forced to choose between community-engaged research and meeting career milestones of frequent peer-reviewed publication and securing research funding. The PROMISE Core fosters investigations in these complex areas while supporting investigators in attaining career milestones.  The PROMISE Core has established a multilevel infrastructure with community stakeholders. The core provides outreach, consultation, training, and technical assistance to investigators regarding effective community-engaged research. Investigators identify and build authentic relationships with community stakeholders, facilitating a context of equal co-participation throughout the research process. The core instills not only the ability for individual investigators to execute projects but also maintains the norm that community-connected work is important across all research stages and at every career phase.  The PROMISE Core is a dynamic, responsive infrastructure that brings together families, community members, researchers, providers, advocates, policy decision-makers, non-profit sector leaders, and other stakeholders in the co-creation of intervention-focused research that addresses the broad goals of addressing pediatric health disparities.
Delaware Center for Neuroscience Research Cell Electrophysiology Core
Cameron Grover* (Delaware State University), Jianli Sun (Delaware State University), Melissa Harrington (Delaware State University)

The Cell Electrophysiology Core lab at Delaware State University is looking for new collaborators. The primary objective of the Electrophysiology Core is to provide critical collaboration for neuroscience projects across University of Delaware (UD), Delaware State University (DSU) and the Nemours A.I. DuPont Children’s Hospital that require analysis of ion channel, synaptic, and network activity in neurons and glia. The Core will perform experimental design, data acquisition, data analysis and assist with result presentation for patch-clamp studies. The Core will also provide training and access to microelectrode array (MEA) system for users to perform their own experiments.
The Delaware Biotechnology Institute Bio-Imaging Center: A highly accessible core facility for advanced microscopy
Tim Chaya, Sylvain Le Marchand, Shannon Modla, Deborah Powell, Jean Ross, Chandran Sabanayagam, and Jeffrey Caplan*

The Delaware Biotechnology Institute Bio-Imaging Center is a multi-user core facility with state-of-the-art microscopy instrumentation, including transmission and scanning electron microscopy, atomic force microscopy, and advanced light microscopy. Some of the newer technologies include an Andor Dragonfly microscope capable of spinning disk confocal microscopy and single molecule localization-based super resolution techniques, a Zeiss CellDiscoverer7 automated high-content imaging system, and a new Stellaris 8 tauSTED super resolution microscope. For electron microscopy, the Bio-Imaging Center just installed a new Thermofisher Talos L120C 120 transmission electron microscope (TEM) capable of cryo-TEM and a Thermofisher Apreo VS scanning electron microscope (SEM) capable of volumetric serial block face SEM. Six full-time staff members are available to advise researchers on instrumentation, sample preparation, experimental design and data analysis and interpretation. The center has a full sample preparation laboratory and the staff research scientists can either conduct all of the sample preparation or train users to prepare their own samples. The Bio-Imaging Center is open to all academic, non-profit and industrial researchers on a fee-for-service basis. Grant Acknowledgements: The core facility is supported by grants from NIH-NIGMS, including DE-INBRE (P20 GM103446) and DE-COBREs (P20 GM139760, P20 GM113125, P20GM104316) and the state of Delaware. Shared instrumentation grants from the NIH-NIGMS (S10 OD016361, S10 OD030321, S10 OD025165) provided funds for the acquisition of microscopes.
Assessment of the Impact of the K-INBRE Program on Undergraduate Student Research Experiences and Career Outcomes
Sarah E. Velasquez, PhD* (University of Kansas Medical Center); Michael Shi (Cornell University)
The purpose of this quality improvement (QI) assessment was twofold: To determine the
satisfaction of the Kansas IDeA Network of Biomedical Research Excellence (K-INBRE) program
on students’ research experiences, including funding; and to determine the impact of the
program on educational and career outcomes of student participants. The initial phase of this
assessment included the design and distribution of a comprehensive survey targeting past
participants of the program, spanning from 2001 to 2023. A second survey of current students
was distributed focusing solely on feelings about funding. The surveys were administered vis
RedCap and distributed via email. The second phase of this assessment involved conducting a
focused group discussion to gather valuable insights and perceptions from current students
about funding. A comprehensive analysis was employed encompassing both quantitative and
qualitative aspects. Preliminary results suggest that satisfaction of the program is high. The K-
INBRE identified 1417 prior students. Of those students, only 1026 could be contacted at the
time of this poster (72%). Of those contacted, 19.1% responded to the survey. Of the current
138 students, 60 (43.5%) responded to the funding survey, and four of those students
participated in the focus group. Feelings about funding are mixed. Leveraging the insights
obtained from these analyses, we will formulate well-informed recommendations aimed at
enhancing the program.