bluuebunny/crossref_metadata_2025 huggingface dataset:
The Role of the Integrated Stress Response in the Biologic Response of Keratinocytes to UVB Exposure
Authors: Prechtel Taylor J. , Diaz Miguel Barriera , Wek Ron C.
Published: January 2023

Background and Hypothesis:Ultraviolet B (UVB) irradiation from the sun can disrupt normal epidermal keratinocyte function, leading to the development of skin cancer. The integrated stress response (ISR) is a conserved cellular mechanism against environmental stressors, such as UV irradiation, through the phosphorylation of the alpha subunit of the initiation factor eIF2 (eIF2a-P). Previously, we demonstrated that the ISR kinase GCN2 plays a role in the response of keratinocytes exposed to UV irradiation. However, the role of other components of the ISR in UV-induced stress has not been well elucidated. We hypothesize that in addition to GCN2, other components of ISR such as PERK, GCN1, and ZAK1 are involved in apoptotic pathways induced by prolonged UVB exposure.
Methods:Gene-specific knockouts (KO) were generated using CRISPR/Cas9 and validated through immunoblotting. Wild-type (WT) and KO NTERT keratinocytes were irradiated with UVB at 0, 100, 200, and 400J/m2. Six hours post-UVB, cell photomicrographs were taken at 0 and 400J/m2. Measurements for apoptosis were then performed by measuring enzymatic levels of caspase-3.
Results:Measurements of caspase-3 following UVB exposure on WT NTERT, PERK-KO, and GCN2-KO showed no difference between the groups. GCN1-KO had significantly greater caspase-3 activity at 400J/m2, whereas ZAK1-KO had significantly reduced caspase-3 activity at the same concentration. GCN2/PERK double-KO had significantly increased caspase-3 activity at 400 J/m2 compared to the individual KO cells.
Conclusion and Potential Impact:Collectively, our data suggests that the ISR may be involved in the apoptotic responses that ensue in keratinocytes exposed to UVB. How GCN2 and PERK could co-interact with GCN1 or ZAK1 to facilitate apoptotic fate in keratinocytes still warrants further research. Nevertheless, elucidating the role of the ISR in normal keratinocyte response to UV irradiation will allow us to define novel therapeutic targets for the treatment or prevention of UV-related skin malignancies.
Diagnostic Yield of Bronchoscopy in Children with Leukemia or Post Hematopoietic Stem Cell Transplant
Authors: Georgescu Livia , Rahrig April , Montgomery Gregory , Rowan Courtney M.
Published: January 2023

This project was funded, in part, with support from the Indiana Clinical and Translational Sciences Institute funded, in part by UL1TR002529 from the National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
Background: The utilization and diagnostic yield of bronchoscopy with bronchoalveolar lavage (BAL) in immunocompromised children is not well understood. We aim to describe bronchoscopy use and complication rates and to investigate diagnostic yield.
Project Methods: This is a retrospective cohort study of 60 children with leukemia or post hematopoietic cell transplant who had a bronchoscopy with BAL at Riley Hospital for Children between 2017 and 2021. Existing datasets and the electronic medical record were used to collect data on demographics, transplant and diagnosis information, as well as respiratory and bronchoscopy data. Inferential statistics were used. Comparisons were done with regression analysis.
Results: Of the 60 bronchoscopies performed, 48 (80%) revealed diagnostic information: 41 (68%) identified a pathogen, 14 (23.3%) found significant secretions/mucus plugging, and 6 (10%) found pulmonary hemorrhage. BAL results led to a change in antimicrobial therapy in 27 (45%) cases. Blood cultures, respiratory viral panels and serum pathogen testing did not always correlate with BAL results. In patients with other infectious workup, 53% of bronchoscopies found a new infectious diagnosis. Of this cohort, 39 (65%) received supplemental oxygen for a median of 3 (IQR: 1-7) days prior to the bronchoscopy. Most patients, 58 (96.7%), had respiratory symptoms for a median of 5 (IQR: 2-18) days prior to bronchoscopy. Antibiotic use prior to bronchoscopy was common with use in 56 (93.3%) patients for a median length of 6 (IQR: 3-14) days. None of these factors were associated with the diagnostic yield. No patient required an escalation of respiratory support post-bronchoscopy. For the 25 (41.7%) children on mechanical ventilation when the bronchoscopy was performed, there was no difference in ventilator settings pre and post-bronchoscopy.
Conclusion and Implications: Bronchoscopies with BAL are useful, safe, and important in the diagnostic management of pulmonary complications in this cohort of children.
Imaging Ligands Targeting Glypican-3 (GPC3) Receptor Expression in Hepatocellular Carcinoma
Authors: Grega Shaun D. , Zheng David Z. , Zheng Qi-Huang
Published: January 2023

Hepatocellular carcinoma (HCC) is the third leading cause of cancer mortality and is increasing in incidence worldwide. Early detection of HCC is important since potentially curative therapies exist in the initial stages of HCC; no curative therapies exist for late-stage HCC. However, the initial detection of HCC remains challenging due to the lack of symptoms during the early stage of the disease. Other methods of screening and detecting HCC, including blood serum tests and conventional imaging methods, remain inadequate due to genetic differences between patients and the high background activity of liver tissues. Thus, there is a need for an accurate imaging agent for the diagnosis, staging, and prognosis of HCC. Glypican-3 (GPC3) is an oncofetal receptor responsible for regulating cell division, growth, and survival. GPC3 is a clinically relevant biomarker for imaging and therapeutics, as its expression is HCC tumor-specific and absent from normal and other pathological liver tissues. The development of novel GPC3-targeting imaging agents has encompassed three classes of biomolecules: peptides, antibodies, and aptamers. These biomolecules serve as constructs for diagnostic imaging (demonstrating potential as positron emission tomography [PET], single-photon emission tomography [SPECT], and optical imaging agents) and HCC treatment delivery. At least 20 unique ligands have been identified in the literature as showing specificity for the GPC3 receptor. Although several ligands are currently under clinical investigation as therapies for HCC, clinical translation of GPC3-targeting ligands as imaging agents is lacking. This review highlights the current landscape of ligands targeting GPC3 and describes their promising possibilities as imaging agents for HCC.
Psychosocial Factors in Chronic Wound Patients
Authors: Gummalla Bharat , Singh Kanhaiya , Sen Chandan K.
Published: January 2023

Background/Objective: Chronic wound (CW) represents a major burden on the individual patients who are typically challenged with many underlying health related complications. Stress is now recognized as a universal premorbid factor associated with many risk factors of various chronic diseases. This study utilized socio-demographic data of CW patients and analyzed it against salivary cortisol, a known marker of psychosocial stress.
Methods: CW patients were recruited under IRB approved protocol of Indiana University (ClinicalTrials.gov Identifier# NCT03793062). Morning saliva samples were analyzed from 11 patients with CW (56.4±2.8 years). The Patient Health Questionnaire (PHQ-9) and Generalized Anxiety Disorder (GAD-7) assessment were used as screening tools as well as measures of symptom severity for depression (PHQ-9) and different types of anxiety (GAD-7). Human salivary cortisol was measured by ELISA.
Results: The salivary cortisol concentration for CW patients were as follows: those with minimal depression (PHQ-9 score ≤4) had 2.2±0.5 ng/mL; those affected by moderate severe depression (PHQ-9 score >4) showed 1.3±0.3 ng/mL; those with minimal anxiety (GAD-7 score ≤4) was 1.8±0.5 ng/mL; and those with mild to severe anxiety (GAD-7 score >4) had 1.6±0.3 ng/mL. Comparison across race showed that the African American CW patients had higher (2.8±0.5 ng/mL) salivary cortisol levels compared to white CW patients (1.5±0.3 ng/mL).
Conclusion and Impact: These preliminary study results indicated that although higher scales of depression and anxiety do not seem to increase salivary cortisol, it is likely that African American CW patients are subject to higher psychosocial stress compared to Caucasian CW patients. The study of a larger sample size is warranted.
Extracellular DEK Treatment Increases Mitochondrial Dysfunction in the Mouse AML Cell line MLL-AF9
Authors: Aguilar Neuville Paola , Ropa James , Gutch Sarah , Capitano Maegan
Published: January 2023

Acute myeloid leukemia (AML) is the most common kind of acute leukemia and the second most common type of leukemia in adults. Poor outcomes resulting from AML are thought to occur due to the inability to target the small pool of cancer-initiating cells that develop from cells within hematopoietic stem (HSC) and progenitor (HPC) cell compartments in bone marrow. DEK, a nuclear protein that can be secreted under stress conditions, plays a role in regulating HSC and HPC function. Extracellular DEK has been found to improve functional HSC expansion in in-vivo and ex-vivo mouse studies. Moreover, RNAseq experiments suggest that recombinant human DEK treatment causes the upregulation of the antioxidant gene programs. Indeed, DEK treatment reduces total reactive oxygen species (ROS) in human umbilical cord blood HSCs and HPCs. Thus, extracellular DEK enhances normal HSC function through antioxidant programs, but the role of the extracellular DEK in AML is unclear. We hypothesized that recombinant mouse (rm)DEK treatment of the mouse-derived AML cell line MLL-AF9 would affect mitochondrial function since mitochondria are an important source of ROS production. Since ROS production can contribute to mitochondrial dysfunction by causing damage to the organelles, we investigated the effects of DEK signaling on mitochondrial metabolism in MLL-AF9 cells using the Seahorse XF instrument, which can measure changes in metabolic flux. Compared to vehicle-treated control, cells treated with DEK demonstrated a decrease in basal and maximal mitochondrial respiration, proton leak, and non-mitochondrial oxygen consumption. Experiments to explore the effects of DEK treatment on the glycolytic function of MLL-AF9 cells are ongoing. Our data shows DEK treatment of MLL-AF9 cells alters mitochondrial function. In the future, we wish to investigate DEK’s effect on proliferation and colony formation.
Analysis of the Population and Distribution of Minorities in the State of Indiana that Utilize Dermatologic Care in the IU Health System
Authors: Arrocha Diana , Somani Ally-Khan
Published: January 2023

Background:
Medical centers require reevaluations to appropriately adapt to everchanging societal needs. This study aims to disclose the extent to which IU Health (IUH) patient demographics represent Indiana’s population. U.S. Census Bureau shows Indiana has had a 42.2% increase in its Hispanic population from 2010 to 2020. Via Cerner, thousands of patient demographics were obtained from the University Hospital (UH) and 4 other IUH dermatologic satellite locations – Zionsville, West, Meridian Crossing (MX), and Methodist – to assess whether patients who visit these locations accurately embody the drastically changing state population.
Methods:
Cerner’s “Kitchen Sink” tool generated a list of patients that had made an appointment at one of the 5 dermatological centers. The search was refined to 2021 and included ethnicity, preferred language, residential zip code, etc. This data was then cross- referenced with Indiana’s Public Data, where 92 counties were grouped into 6 regions.
Results:
Indiana’s Public Data for 2021 revealed that 8.3% of the population identified as Hispanic. These Hispanics were localized into residential regions, with the majority being in region 1 (40%), followed by region 4 (29%), region 2 (10%), region 3 (9%), regions 5 (5%) and 6 (5%). Our analysis revealed that 0.6% of Hispanics in Indiana utilized an IUH dermatology facility. Interestingly, despite most Hispanics residing in region 1, only a total of 4% were seen by IUH (UH & MX).
Conclusion:
Our preliminary findings show most Hispanics utilized an IUH facility in region 4, where they resided. Furthermore, we did not see any statistically significant differences in site preference within this group. However, we were surprised by the low number of Hispanics from other regions, especially region 1, utilizing IUH dermatologic care. This study raises the question of potential barriers (proximity, preferred language, lack of awareness, and need for dermatology services) to dermatologic care available to Hispanics within the IUH system. Further studies are needed to identify the cause and potential solutions to the discrepancies within the Hispanic community in Indiana and IUH Dermatology.
Therapeutic Effects of Benzoylacetonitrile on Microglia Activation in Multiple Sclerosis
Authors: Zhao Zixuan (Angela) , Kuo Ping-Chang , Scofield Barbara A. , Yen Jui-Hung Jimmy
Published: January 2023

Background:
Multiple Sclerosis (MS) is an autoimmune disease of the central nervous system (CNS). Pathogenic T cells, such as Th1 and Th17, infiltrate the CNS, resulting in neuroinflammation, demyelination and axonal damage. Th1 activates microglia (MG) in the CNS and Th17 acts as a chemokine to recruit immune cells into the CNS. MG is a resident immune cell in the CNS and its activation is associated with destruction of myelin and secretion of inflammatory cytokines such as IL-12, IL-23 and IL-1β. IL-12 and IL-23 are important for Th1 and Th17 differentiation and reactivation, respectively. IL-1β is a key mediator of the inflammatory response. Benzoylacetonitrile (BTN) has been shown to reduce disease severity in mouse model of MS and reduce Th1 and Th17 differentiation in vitro. However, the effects of BTN on MG are unknown, and this study was aimed to investigate the effects of BTN on MG activation in vitro. We hypothesize that BTN can suppress MG activation and decrease the production of inflammatory cytokines.
Methods:
Primary MG were pretreated with BTN at concentration of 200μM or 300μM for 2 hours or with DMSO (vehicle), followed by lipopolysaccharide (LPS) 100ng/ml stimulation for 1.5 or 3 hours. RNA was isolated from MG and mRNA expression levels of IL-12, IL-23, IL-1β were measured using Q-PCR.
Results:
Our results showed that BTN suppressed MG activation and reduced inflammatory cytokine production. The mRNA expression levels of IL-12, IL-23, and IL-1β in LPS and BTN-treated MG were significantly lower than LPS-treated MG.Conclusion: This study demonstrated that BTN was able to suppress MG expression of inflammatory cytokines in vitro, suggesting that BTN exhibits immunomodulatory effects on MG activation in vitro. BTN has a potential to attenuate neuroinflammation in MS through the reduction of inflammatory cytokines.
Conclusion:
This study demonstrated that BTN was able to suppress MG expression of inflammatory cytokines in vitro, suggesting that BTN exhibits immunomodulatory effects on MG activation in vitro. BTN has a potential to attenuate neuroinflammation in MS through the reduction of inflammatory cytokines.
Evaluation of Cost and Parental Satisfaction in Type B Polydactyly Excision Completed in Office Vs. Operating Room
Authors: Yun EJun , Tran Khoa , Borschel Gregory H.
Published: January 2023

Background/Objective:One in 500 children in the US are born with polydactyly, extra digits on hands or feet. Out of those children, 95% of cases comprise of type B polydactyly, in which the extra digit is attached by neurovascular bundle without bones. Surgeries to remove type B polydactylies by excision have been traditionally performed in operating rooms. Even though operating room surgeries are successful, they can be expensive and time consuming. In contrast, the surgeries performed in physician’s office, while non-traditional, can minimize expenses and save time and effort. We hypothesize that excisions conducted in offices are safe alternatives to operating room excision and are associated with definitive financial savings for patients’ families and hospitals, while increasing satisfaction of parents.
Methods:Using CPT codes and reviewing electronic medical records, we identified all patients from 2019 to 2022 who had been treated for Type B Polydactyly with either office procedures or OR procedures, all of which had been performed at Riley Children’s Hospital. Patient billing data were obtained from Indiana University Hospital billing department. Average charge to patients for total cost of procedure and estimated removal of one digit were analyzed and compared. A 9-question parental satisfaction questionnaire was administered to patient’s parents over phone call.
Results:Thus far, out of 93 patients who underwent type B polydactyly removal, 36 patients received procedures in physician’s offices, and 57 patients received procedures in ORs. Average charge to patients per digit removed for in-office group was around $500 cheaper than that of OR group. Average total charge to patients for in-office group was around $600 cheaper than that of OR group. Parental satisfaction data are pending.
Conclusion and Potential Impact:This study shows the cost effectiveness of type B polydactyly excision in office. With analysis of future satisfaction survey data, we hope to gauge parental satisfaction associated with in-office excisions and promote in-office excisions as effective alternative to OR excisions.
Direct Inspection of Primary Aortic Cell Transcriptomes Identifies Candidate Causative Variants in Patients with Thoracic Aortic Aneurysm
Authors: Mederos Alexa V. , Landis Benjamin J.
Published: January 2023

Background: Thoracic aortic aneurysm (TAA) is an aortopathy that predisposes to life-threatening aortic dissection. Autosomal dominant disorders associated with TAA include Marfan syndrome (FBN1), Loeys-Dietz syndrome (TGFBR1/2, SMAD3, TGFB2), and vascular type Ehlers-Danlos syndrome (COL3A1). Our objective was to identify single nucleotide variants (SNVs) in these six genes within the transcriptomes of primary aortic cells acquired from patients with aortopathy.Methods: Primary aortic cell lines were cultured directly from the medial layer of surgically explanted aortic tissues in 63 unrelated aortopathy patients. RNA samples were extracted from aortic cells for mRNA sequencing. RNA reads aligning to the 6 selected TAA genes were directly inspected using Integrative Genomics Viewer (Broad Institute), and the identified SNVs were filtered for downstream analysis.Results: Study patients were predominantly male and of European ancestry with a mean age of 52±18 years. Thirty-three (52%) patients had a bicuspid aortic valve, and 10 (16%) had family history of TAA or dissection. A total of 3740 SNVs were identified in patient transcriptomes, and these occurred at 905 distinct genomic coordinates. There were 115 SNVs that were unique within the cohort, not located within a 3’-untranslated region, and had ≥20 aligned reads at the SNV’s position. Using the application Franklin (Genoox) to estimate clinical interpretation, 8 unique SNVs were classified as pathogenic (P) or likely pathogenic (LP). Five of these P/LP SNVs were associated with reduced allelic expression, and gene expression level was below the 20th percentile of study samples for 6 P/LP SNVs. Eighty-six unique SNVs were classified as variants of uncertain significance (VUSs). A total of 39 patients (62%) had at least one SNV classified as P/LP/VUS.
Conclusion: Transcriptomic analysis of primary aortic cells identified candidate causative SNVs and their relative allelic expression. Further analyses will investigate additional TAA-associated genes and integrate transcriptional abnormalities with genetic variants.
Targeting Arg-1 and PD-L1 in M2-Tumor Associated Macrophages Impairs Juvenile Myelomonocytic Leukemia (JMML) Cell Proliferation and Migration
Authors: Young Kailah , Pasupuleti Santhosh Kumar , Stieglitz Elliot , Kapur Reuben
Published: January 2023

Background and Hypothesis: Tumor-associated macrophages (TAMs) are a key component of tumor-infiltrating immune cells. They are largely characterized into M1 or M2 types. TAMs express an anti-inflammatory M2-like phenotype, promote tumor progression. However, the role of M2-TAMs in driving disease pathogenesis in patients with Juvenile myelomonocytic leukemia (JMML), a rare form of pediatric leukemia driven to a large extent by mutations in the PTPN11 gene, which encodes the phosphatase SHP2 is unclear. We hypothesized that in JMML, inflammatory myeloid cells including neutrophils and M2-TAMs express higher levels of arginase-1 (Arg-1) and PD-L1, which may contribute to the local suppression of immune responses and support the development of JMML.
Methods: To study how alterations in M1/M2 macrophages contribute to JMML development, we utilized a mouse model bearing Shp2E76K mutation (Ptpn11E76K/+) which manifests the cardinal features of human JMML. We hypothesized that Shp2E76K/+ mutations enhance the function of bone marrow derived macrophages (BMDMs), including M2-TAMs and contribute to T-cell suppression.
Results: Our analysis of the bulk RNA-sequence data from 90 JMML patients showed increase in the expression of Arg-1 and PD-1. Furthermore, single cell RNA-seq analysis of macrophages from 4 JMML patients revealed higher expression of M2-macrophage markers/genes. Our results show that in M2-TAMs, Arg-1 and PD-L1 levels are elevated in BM and spleens of Shp2E76K/+ mice compared to WT. Moreover, M2-TAMs, Arg-1 and PD-L1 levels were also higher in BMDMs derived from Shp2E76K/+ mice compared to WT. The BMDMs from Shp2E76K/+ mice have greater proliferation and migration potential compared to WT BMDMs, which was significantly reduced by inhibiting the function of Arg-1 and PD-L1.
Conclusion: Our results show that M2-TAMs, arginase-1, and PD-L1 create a pro-tumor microenvironment, which likely contributes to the growth of JMML cells. Inhibition of Arg-1 and PD-L1 is a novel therapeutic approach to treat patients with JMML.
Assessing Continuous Glucose Monitor Alarm Use by Families of Children with Diabetes Seen at Riley Children’s Hospital
Authors: Ochs Victoria , Woerner Stephanie , Hilliard Marisa , Ustyol Ala , DiMeglio Linda
Published: January 2023

Background/Objective:A useful feature of continuous glucose monitors (CGM) is customizable alarms which alert persons with diabetes and their caregivers of current or pending blood sugar changes. However, there has been little work studying real-world CGM alarm settings and use in large clinical populations. We seek to understand CGM alarm use through analyses of CGM reports and user interviews.Methods:We analyzed data from two-week CGM reports obtained clinically to determine common alert/alarm settings and compared those to target blood sugar guidelines. We also studied families who are currently using or have never used CGM for diabetes management of their children aged 2-18 years through standardized surveys about diabetes technology and qualitative interviews investigating benefits and burdens associated with CGM use. Data were tabulated, and qualitative analyses were performed.Results:CGM download data from 140 children using the Dexcom® G6 was analyzed. The Low alarm was used by 122 participants (87%) with a median blood glucose of 74 mg/dL (range: 60-100) used as a threshold for alert. The High alarm was used by 101 participants (72%) with a median blood glucose of 274 mg/dL (range: 120-400) used as a threshold for alert. Low Repeat and High Repeat alarms were used by 48 (34%) and 35 (25%) participants, respectively. The most common themes revealed during interviews include the alarms were helpful during sleep, a low sugar is the most important information to be alerted about, and high sugar alarms can be disruptive to patients and caregivers.Conclusions and Potential Impact:We anticipate that identifying barriers to using CGM alarms will assist providers in efforts to instate customizable alerts and alarms. Setting CGM alarm best practices will help ensure children with diabetes and their caregivers are getting needed health data to optimize care and minimize feeling overwhelmed by constant alarms.
Grounded Practical Theory Analysis of Patient-Provider Communication with Black Women Participating in Breast Cancer Clinical Trials
Authors: Okoruwa Oseme P. , Ridley-Merriweather Katherine E.
Published: January 2023

Background:Previous literature suggests breast cancer clinical trial participation among Black women has declined in recent years by as much as 35%. Though the literature identifies barriers to participation for this population, little has been studied about how researchers can address these barriers. This study investigates the communication between healthcare providers and Black women to reveal how providers and researchers can positively influence their perceptions of breast cancer clinical trial participation.Methods:Fourteen women (n=14) who self-identified as Black, Black American, or African American, were interviewed about their communication experiences with healthcare providers regarding breast cancer clinical trial participation. Each transcribed interview was coded using thematic analysis. Grounded Practical Theory was introduced to give insight into the patient-provider communication needs of Black breast cancer research participants.Findings:The findings fell into four categories: (1) impressions of participants toward their providers, (2) reflections on the clinical trial recruitment experience, (3) communication relationships with medical and research providers, and (4) cultural aspects of patient-provider communication. One major finding was that an important way women learn about clinical trials is through conversations with their oncologists. However, only 29% of Black women interviewed were informed of their clinical trial by a healthcare provider, suggesting that Black women may not be receiving the information they need to participate in clinical trials.Conclusion:By understanding existing patient-provider communication typologies, we can improve these methods of communication to increase the interest and participation of Black women in breast cancer clinical trials.Implications:Clinical trials provide data to healthcare providers about treatment options for breast cancer. If minoritized populations are continually underrepresented in clinical trials, these treatments might not prove to be efficacious in Black women. Researchers must make the necessary investment of resources and effort to better understand the needs of Black women in clinical trial recruitment.
Is β-Cell Dysfunction Present in Adult Autoantibody Negative Relatives of Individuals with Type 1 Diabetes Mellitus?
Authors: Pacheco-Figueroa Adriana , Monaco Gabriela S. F. , Neyman Anna , Sims Emily K.
Published: January 2023

Background/Objective: Individuals with a family history of type 1 diabetes mellitus (T1D) are at increased genetic risk for T1D. Previous studies identified the presence of β-cell dysfunction before clinical onset and diagnosis of T1D. However, it is unclear if β-cell dysfunction predates islet autoimmunity in individuals at high genetic risk. Our objective was to test β-cell function in islet antibody negative adults who have a first-degree relative with T1D. We hypothesized that individuals at genetic risk for T1D would exhibit β-cell dysfunction even without detectable islet autoimmunity.Methods: We used ordinary one-way and Brown-Forsythe ANOVA to compare the repeated mixed meal tolerance test (MMTT) and hyperglycemic clamp glucose-stimulated β-cell response and function measures between three groups of individuals: normoglycemic adults without T1D family history age, sex, and BMI-matched islet antibody negative first-degree relatives of individuals with T1D, and islet antibody positive first-degree relatives of individuals with T1D.Results: Neither the MMTT first-phase insulin secretion measures (c-peptide0-15 minutes, c-peptide0-30 minutes, insulin0-15 minutes, insulin0-30 minutes), nor second-phase measures (c-peptide0-120 minutes, insulin0-120 minutes, and glucose0-120 minutes) showed a statistically significant difference between groups. The clamp acute c-peptide response to glucose, insulin sensitivity, c-peptide steady state, first-phase β-cell function, and second-phase β-cell function were similar between subject groups in both visits. Fasting proinsulin:c-peptide ratios, a biomarker of β-cell stress, were also similar between participant groups.Conclusion and Impact: Our data suggest that genetically at-risk autoantibody negative adult relatives of individuals with T1D do not demonstrate β-cell dysfunction compared to controls. Studies show that β-cell ER dysfunction preceding T1D onset is more striking in younger children. Thus, our findings may reflect the use of an adult study population. Alternatively, β-cell dysfunction in T1D may require initial autoimmune activation. This study will contribute to the growing understanding of risk factors contributing to T1D development.This project was funded, in part, with support from the Research Training Program in Diabetes and Obesity funded, in part by grant 3T32DK064466 from the National Institutes of Health. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Funding also provided by JDRF grant 2-SRA-2017-498-M-B.
Histologic Diversity of Thymic Epithelial Tumors in Patients with Myasthenia Gravis
Authors: Heldman Emily M. , Davis Hannah O. , Laniak Louis J. , Wuthrich Brice S. , Loehrer Patrick J. , Kesler Kenneth A.
Published: January 2023

Background and Objective:Thymic epithelial tumors (TETs) include thymic carcinomas and thymomas, the latter of which can be further categorized by the World Health Organization (WHO) histologic classification based on the morphology of epithelial cells and the ratio of lymphocyte to epithelial cells. TETs are rare malignancies with an incidence of 0.15 per 100,000 in the United States. While their etiologies remain unknown, these tumors are associated with distinctly high rates of autoimmune disorders and paraneoplastic syndromes. Approximately 30% of patients with thymoma also carry a diagnosis of myasthenia gravis (MG). For the present retrospective study, we created a database of patients with TETs and examined prevalence of each histologic subtype in patients with MG.
Methods:Drs. Patrick Loehrer, Kenneth Kesler, and colleagues have collaborated at the Indiana University Simon Comprehensive Cancer Center to care for over 1000 patients with TETs. The electronic health records of these patients were accessed via Cerner and used to input demographic, diagnostic, and histologic data into a REDCap database. These tumors were further categorized by WHO classification, and those comprising multiple histologic types were included separately in each subgroup.
Results:Of 626 patients deemed to have sufficient documented histologic and diagnostic data, 110 had both MG and a TET diagnosis confirmed by pathology report. The greatest prevalence of WHO histologic type in patients with thymoma and MG was Type B2 (47), followed by Type B3 (34), Type B1 (22), Type A (9), Type AB (7), and unspecified thymoma (7). Notably, only 3 of 159 (1.87%) total patients with thymic carcinoma had comorbid MG in contrast to 107 of 467 (22.91%) patients with thymoma and MG; this suggests a uniquely favorable microenvironment of thymoma in patients with MG.
Clinical Impact and Implications:Future work will aim to determine whether histologic classification has a predictive value for tumor prognosis in patients with and without MG. Furthermore, patterns of gene expression associated with thymoma in patients with and without MG may elucidate the etiologic mechanisms for the development of this autoimmune disorder.
Comparison of Chiropractic Manipulation Therapy and Functional Movement-Based Myofascial Release in Shoulder Range of Motion of Collegiate Athletes: A Pilot Study
Authors: Heumann Ryan , Waltz Matt , Garcia-Hosokawa Marcel , Chlebowski Arthur L.
Published: January 2023

Background: Shoulder pain is the third most common presentation of musculoskeletal pain in the clinic with a lifetime prevalence of up to 70%. In athletes, shoulder dysfunction is often due to the extreme forces experienced by the joint during sport participation. Studies have shown that a deficit of 5 degrees in total arc of motion, 20 degrees in internal rotation, and 15 degrees in horizontal adduction can increase an athlete’s chances of injury by a factor of four.
Methods: Student athletes with shoulder pain and decreased range of motion (ROM) in their dominant arm were separated into cohorts and received twelve therapy sessions. One cohort received chiropractic manipulation (CM) and the other received functional movement-based myofascial release (FMMR) with CM. ROM of the shoulder was measured during the first, sixth, and last sessions using goniometry and video capture. Cohorts were compared using statistical analyses on data collected.
Results: A total of four participants have completed at least the first six sessions of participation in study. Initial results show there is no statistical difference in improvement in shoulder ROM between cohorts. Student T-tests comparing cohorts’ improvements in six of the seven specific ROM tests resulted in no statistical significance (p-values ≤ 0.05). The two participants that completed all twelve intended therapy sessions showed evidence of positive trajectory for increased ROM and qualitatively expressed improvement in shoulder motion.
Conclusion: This study shows that there was no statistically significant difference in treating athletes with CM versus FMMR combined with CM. This can mainly be attributed to the study’s current size (n=4). Even with the small participant size, and lack of statistical significance several trends of the individual range of motion measurements provide questions that would benefit from the continuation of the study towards the full cohort participation.
Cochlear Explantation Prevents Hair Cell Degeneration in Transmembrane Serine Protease 3 (Tmprss3) Deficient Mice
Authors: Padgett Lauryn , Chen Jeffery , Cabrera Ernesto , Nelson Rick F.
Published: January 2023

Background/Objective: Transmembrane serine protease 3 (TMPRSS3) is the most common causative hearing loss gene in adults undergoing cochlear implantation and a significant cause of non-syndromic hearing loss. However, the function of TMPRSS3 within the inner ear is unknown. In murine models, Tmprss3Y260X/Y260X mutants (Tmprss3-/-) have normal hair cell development followed by rapid hair cell degeneration from postnatal day 12 (P12) to P14. The endocochlear potential in vivo rises from P7 to a peak at P12, when mice begin to hear, which temporally corresponds to the HC degeneration in Tmprss3-/- mice. We tested if hair cell death occurs after removal of the endocochlear potential through cochlear explantation.Methods: P7 organ of Corti explants from control (Tmprss3+/-) and Tmprss3-/- mice were cultured in 5 mM potassium solution for 7 days in vitro (equivalent to P14). Whole mounts of cultured (P7+7DIV) and in vivo P14 cochlea were immunostained for MYO7A (hair cell marker) and DAPI followed by quantification of inner and outer hair cell (IHC/OHC) counts per 20 μm. Statistical analysis included two tailed t-test with p-value of (<0.01, n=6).Results: As expected, there is complete loss of hair cells at P14 in vivo in Tmprss3-/- mice. Compared to P14 in vivo, Tmprss3-/- OC explants displayed significantly improved IHC and OHC survival (P<0.001). IHC and OCH survival was similar between control and Tmprss3-/- OHC explants in vitro (P=0.99, n=6)Conclusion: These results suggest that degeneration of Tmprss3-/- hair cells is due to factors related to the endocochlear potential and implicate TMPRSS3 function in regulation of epithelial tight junctions. Future directions include confirming that hair cell death is potassium-mediated by crossing Tmprss3-/- mice with Pou3f4del-J mice, which have a decreased endocochlear potential without hair cell degeneration.
Continuous Glucose Monitoring Workflows for Elderly, Cognitively Impaired Adults with Type 2 Diabetes
Authors: Pamidimukkala Ujwala , Savoy April
Published: January 2023

BackgroundElderly adults with Alzheimer’s disease-related dementia (ADRD) and type 2 diabetes tend to have difficulty in detecting hypoglycemic events. Over time, recurring hypoglycemic events increase the risk for severe consequences, such as hospitalization. Previous studies have shown continuous glucose monitoring (CGM) systems to be one of the best predictors of hypoglycemia, which can be difficult to discern normally. However, CGM systems have not been formally introduced to the ADRD population, so there is a need to understand how CGM can be incorporated into the diabetes care of elderly, cognitively impaired adults.ObjectiveThe goal of this project is to develop a better understanding of how CGM systems can be extended to the ADRD population and what potential barriers may develop.MethodsA narrative review of how CGM systems are currently used by patients and caregivers was conducted using databases such as PubMed and Google Scholar, as well as clinical and CGM manufacturer manuals. Subsequently, a workflow extrapolated to ADRD adults was created based on these sources.FindingsA total of 118 articles, websites, and guides were obtained and evaluated. Current CGM workflows consist of 3-9 steps. A total of five potential areas for improvement have been identified. The newly constructed workflow consists of 9 steps: (1) healthcare visit, (2) CGM education, (3) CGM pick-up, (4) sensor insertion, (5) scan/calibrate, (6) evaluate data, (7) replace sensor, (8) next healthcare visit, and (9) pharmacist alterations.ConclusionCurrent CGM workflows are oversimplified and do not detail processes that can be complicated for adults with diabetes and ADRD and their caregivers. However, more research still needs to be conducted to determine the severity of the identified barriers and how to overcome them. This project can inform future work on the integration of CGM into diabetes care for the ADRD population.
Evaluation of Chemotherapeutic Outcomes for Thymic Carcinoma Patients
Authors: Wuthrich Brice , Davis Hannah , Laniak Louis , Heldman Emily , Loehrer Patrick J. , Kesler Kenneth A.
Published: January 2023

Background and Hypothesis:Thymic Epithelial Tumors are uncommon tumors of the anterior mediastinum composed ofthymomas and thymic carcinomas (TC). TC’s are known to have worse disease outcomes andlower rates of survival in comparison to thymomas and are suspected to have lower responserates to chemotherapy as well. As these tumors are rare, little data exists assessing the trueefficacy of chemotherapeutic regimens for TC patients. Due to this lack of data, and that theIndiana University Health Simon Cancer Center treats a high percentage of TET patients, adatabase of these patients covering a variety different disease characteristics and treatmentshas been established. We hypothesize, upon evaluation of this database, response rates toanthracycline based regimens (PAC) will be superior to non-anthracycline based regimens (i.e.,PE, Carbo/Taxol) for TC patients.
Methods:In this project, a collection of patients seen by Dr. Patrick Loehrer and/or Dr. Kenneth Keslerwas acquired, and a database was created using these patients in RedCap. Once established,we evaluated patient medical records in Cerner and entered data related to diseasecharacteristics and treatments. These patients were then analyzed accordingly to evaluatechemotherapy response rates.
Results:The database yielded 123 instances of chemotherapy treatment for TC. Of which, the mostpopular treatments were PAC and Carbo/Taxol. The data suggests that PAC generates a higherresponse rate (65.5%) than other therapies (Carbo/Taxol: 27.6%, PE: 58.3%, etc.). Therefore,there is evidence that anthracycline based regimens may be more effective at generatingresponse rates in comparison to non-anthracycline based regimens.
Conclusion and Potential Impact:This project will help elucidate the effectiveness of recommended systemic therapies for thymiccarcinoma patients from one of the largest TET databases constructed. Ultimately, we hope thatwith clarity of the effectiveness of treatment, this can serve as a reliable reference for evidencebasedmedicine for the care of TC patients.
Challenges of Recruitment of Diverse Populations in Clinical Research
Authors: Hannah Hannah , King Carli , Hoffman Shelley , Tucker Edmonds Brownsyne
Published: January 2023

Objective: The recruitment of diverse populations in research is essential for representing marginalized groups. We discuss the successes and challenges of recruiting pregnant-parent dyads for a study examining neonatal decisional authority within diverse partnerships.
Methods: We aimed to recruit 30 adult, first-time pregnant people between 18-26 weeks gestation, along with their partner. Heterosexual, same-sex, married, and unmarried partnerships were recruited to ensure a sample of diverse partnerships. Study advertisements were shared on pregnancy-related Reddit and Facebook groups. Flyers were distributed to OB/GYN clinics, LGBTQ+ healthcare providers, public events, and coffee shops. In-person recruitment at local clinics began in July 2022. Ads included a hyperlink or QR code to direct individuals to a brief screening questionnaire. If deemed eligible, the research team contacted the candidate to confirm eligibility and to schedule interviews. Virtual interviews lasted 1-hour and each participant received a $50 gift card.
Results: A total of 88 (51.4%) individuals were initially eligible. Of those, 35 (39.8%) did not leave contact information and 23 (26.1%) did not respond. To date, 20 parent dyads have been interviewed, with most recruited from Reddit (n=17, 85%). The majority of dyads were white. Four (10%) participants were non-white and four were Hispanic/Latinx. Additionally, 4 (10%) participants identified as LGBTQIA+. All eligible dyads were married. Challenges to recruitment include lacking access to private social media groups, a high prevalence of scammers, unresponsiveness, and difficulty getting buy-in from clinicians to share study ads with patients. In-person recruitment is promising but difficult to coordinate and limits the study’s generalizability.
Conclusions: Online recruitment provides potential for recruiting diverse populations but yields several challenges. This presentation can provide insight for studies needing to recruit these populations.
Analysis of Neutral Lipid Accumulation and Lipotoxicity in Non-hepatocyte Liver Cells Under Free Fatty-acid Treatment
Authors: Wilmes Danielle , Park Yujin , Farag Kristine , Li Ping , Zhang Wenjun , Ekser Burcin
Published: January 2023

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) and its advanced form, nonalcoholic steatohepatitis (NASH), are prevalent liver diseases with no effective treatment available. NAFLD is marked by increased lipid accumulation in the hepatocytes, lobular inflammation and fibrosis. However, the effects of lipid accumulation in other hepatic lineage cells, such as liver endothelial cells (LEC), intrahepatic cholangiocytes (IHCHOLs), and hepatic stellate cells (HSCs), on NAFLD/NASH progression are less understood.
METHODS: Human HSCs, IHCHOLs, and LECs were isolated from liver explants of healthy donors and further characterized with immunofluorescence staining. The cells were treated with different concentrations of free fatty acids (FFAs, oleic acid and palmitic acid 2:1) for 48 hours, and then examined for lipid accumulation with staining of BODIPY493/503. A cell viability assay and LDH cytotoxicity assay were performed. Real-time PCR was performed to monitor gene expression in LECs after treatment.
RESULTS: Lipid accumulation confirmed by BODIPY493/503 staining was observed following treatment, especially with higher FFA concentrations. No statistically significant difference in cell viability or cytotoxicity of the cells was observed under different treatment concentrations of FFA. However, a negative trend in cell viability was observed with an increment of FFA dosage. FFA treatment upregulated the expression of the genes in LEC that are related to angiogenesis (CDH5), epithelial-mesenchymal transformation (TGF-β2 and TGF-β3), cellular stress (P53), inflammation (IL-32) and immunoglobular cell adhesion molecules (ICAM-1 and VCAM-1) that are key to monocyte adhesion to endothelial cells in response to a local inflammatory signal.
CONCLUSION: Using an in vitro cellular model system, we examined lipid accumulation and cellular stress in different human liver cell types, especially LECs, under dose-dependent FFA treatment. This could be further developed to delineate adaptive responses within different hepatic lineage cells in the progression of NAFLD/NASH.
IMPACT: A better understanding of the involvement of hepatocytes and other hepatic lineages in the progression of NAFLD/NASH will augment the development of targeted and effective treatments.
Comparison of Goniometry and Video Analysis in Measuring Shoulder Range of Motion of Collegiate Athletes
Authors: Waltz Matt , Heumann Ryan , Garcia-Hosokawa Marcel M. , Chlebowski Arthur L.
Published: January 2023

Background: The shoulder joint’s multiplanar motion creates a challenge in measurement of range of motion (ROM). Goniometry is currently the most common tool for measuring ROM in the clinic due to its ease of use, but is subjective in nature. Video capture rigs have become popular in all facets of motion monitoring, but they can be limited to large open spaces and user discrepancy. Further, motion capture with wireless wearable inertial measurement units (IMUs) has also become popular but is often cost/implementation prohibitive. Understanding the consistency between these recording methods must be established to understand if a therapy impacts the shoulder complex.
Methods: Student athletes with complaints of shoulder pain and decreased ROM in their dominant arm were separated into cohorts receiving twelve therapy sessions. One cohort received chiropractic manipulation therapy and the other received functional movement-based myofascial release therapy combined with chiropractic manipulation. Shoulder ROM was measured via seven standard tests during the first, sixth, and last sessions using goniometry, video capture, and IMUs. Following data collection, results of the measurement techniques were compared.
Results: Preliminary statistical analysis conducted between goniometry and video capture indicates measurement techniques differ for three of seven standard ROM tests, pronated flexion, supinated abduction, and internal rotation (p-value ≤ 0.05). Reliability between individuals analyzing video capture is strong, with an ICC of 0.994. This indicates consistency between individuals taking measurement of shoulder ROM using this method. IMU data has been collected with analysis forthcoming.
Conclusion: This study is limited by the number of participants (n=4). Preliminary results indicate that consistency in video analysis may make it preferable to standard goniometry. A larger, more comprehensive study is needed to provide conclusive results, including IMU data. These conclusive results could indicate that clinical use of video capture or IMUs should replace standard goniometry if other implementation hurdles can be reduced.