bluuebunny/crossref_metadata_2025 huggingface dataset:
Prevalence of impairing behavioral health problems in ED patients and association with ED utilization.
Authors: Nader Setarah Mohammad , Musey Jr., MD, MS, FACEP Paul
Published: October 2019

Background and Hypothesis: It has been observed that patients with poor mental health are relatively frequent users of the Emergency Departments (ED). The objective of this study is to evaluate the prevalence of numerous behavioral health domains (depression, anxiety, PTSD, substance abuse, and suicidality) in patients presenting to the Emergency Department and the association of each of these domains with ED utilization.
Experimental Design or Project Methods: This prospective study seeks to enroll a convenience sample of 1000 Englishspeaking adults presenting to IU Health Methodist and Eskenazi Emergency Departments without psychiatric chief-complaints. Patients were assessed for behavioral health problems using the CAT-MHTM, PHQ-8 and GAD-7 screening tools, which were administered via tablet device. Additionally, data on disposition medical history, discharge diagnoses, and ED utilization in the 12 months before and after enrollment from electronic medical records and data from the Indiana Network for Patient Care (INPC) will be reviewed.
Results: Over the course of five weeks, 375 patients have been enrolled. Of those 59.4% were female with an overall mean age of 46.1 (SD ± 16.4); 52.9% were white and 39.8% black/African American. Among enrollees 42.2% screened positive for depression, 29.7% for anxiety, and 1.3% for suicidal ideation. Patients who screened positive for depression were predominately females (76.1% vs 23.9%), those who screened positive for anxiety were also predominately females (71.6% vs. 28.4%). However, 3 out of the 5 (60%) patients that screened positive for suicidal ideation were males. The preliminary analysis of GAD-7 showed of those enrolled 215 (57.5%) had no anxiety, 157 (42%) had mild-severe anxiety. PHQ-8 scores showed 194 (51.9%) had no depression, 178 (47.5%) had mild-severe depression. Similarly, CAT-MH results showed 216 (57.8%) had no depression, 158 (42.2%) had mild-severe depression, while 263 (70.3%) had no anxiety and 111 (29.7%) had mild-severe anxiety. Full data analysis including comparative analysis of the CAT-MH with PHQ-8 and GAD-7 scores will take place after 1000 patients have been enrolled and data has been received from the INPC.
Conclusion and Potential Impact: In our sample, almost half of patients that visit the ED have screened positive for mental health problems. We believe that early identification and appropriate referral may reduce inappropriate ED utilization.
Targeted Elastin-like Fusion Proteins for Enhanced Imaging and Treatment of Non-Muscle Invasive Bladder Cancer
Authors: Nordland Matthew L. , Sweet Craig J. , Shinkle Mollie , Thompson David H.
Published: October 2019

Background and Hypothesis: A recent study estimates 17,670 individuals will die from bladder cancer in the United States in 2019. With current imaging and treatment technology, bladder cancer is associated with high recurrence rates making it the highest lifetime treatment cost per patient. High expense is primarily due to the profound heterogeneity associated with bladder cancer necessitating repeated intervention. Traditional treatment of non-muscle invasive bladder (NMIBC) cancer following intravesical tumor resection often includes chemotherapy or immunostimulatory therapy via Bacillus Calmette-Guerin (BCG) infusions. Both lack specificity for cancerous tissue, leading to sub-optimal prognosis and unnecessary discomfort for patients. Furthermore, bladder cancer detection is limited in its precision. Methods such as CT/MRI urography and cystoscopies are unable to accurately detect tumors under 2 centimeters in diameter. Blue light cystoscopy, a gold standard detection method, is unable to accurately differentiate tumors from inflamed tissue. This leads to issues detecting aggressive carcinoma in situ. The goal of this project is to enhance tumor detection by fusing various targeting ligands to elastin-like peptides (ELP).
Experimental Design or Project Methods: Using synthetic biology approaches, a library of ELP fusions were made and purified using a simplified organic extraction method. (Biomaterials Science 2018, 6 (4), 863-876) This resulted in exceptionally pure protein within a matter of hours. Contrast agents were covalently attached, and performance testing was done in murine bladder cancer cells.
Results: Novel ELP fusions were successfully produced and sequences verified. Protein expression and purification yielded active targeted constructs which showed enhanced binding efficiency over non-targeted constructs. These ELP fusion proteins are effective transporters for rapid and preferential cargo entry into bladder cancer tissue.
Conclusion and Potential Impact: Chemotherapeutic, immunostimulatory, and tumor imaging cargo are all bright candidates for targeted ELP fusion technology. Targeted ELP delivery may be a superior tool key to improving tumor visualization and treatment, leading to improved comfort and prognosis in patients with NMIBC.
The Effect of Chronic Hypoxia on Airway Remodeling in Murine Model of Asthma
Authors: Ozolin Jordan , Gao Amy , Perez Page , Tepper, MD PhD Robert
Published: October 2019

Background and Hypothesis: Chronic hypoxia during growth and development results in adaptive responses to increase oxygen transport to tissues, such as increasing lung volume and lung diffusion. Asthma is characterized by airway hyper-reactivity, which can result from increased airway smooth muscle (ASM) and airway remodeling following airway inflammation. Chronic hypoxia may attenuate airway inflammation, airway reactivity and airway remodeling in patients with asthma. Measured by in vivo airway resistance in response to 50 mg/ml Ach, our preliminary data suggests that rats conceived and raised under chronic hypoxic conditions (15% O2) exhibit lower airway reactivity, compared to room air controls (20% O2) at baseline (0.397, 0.789 respectively) and following Ova sensitization/challenge (0.696, 1.159 respectively). We hypothesized lower airway reactivity was associated with less ASM in response to hypoxic conditions.
Methods: Sprague Dawley rats were conceived, raised and evaluated in 4 groups: hypoxia/PBS challenge (HA-PBS); hypoxia/Ova challenge (HA-Ova); normoxia/PBS challenge (RA-PBS); normoxia/Ova challenge (RA-Ova). The lung tissue (4 animals/group) was fixed, sectioned, and ASM was quantified by immunohistochemistry using imageJ software.
Results: Measured ASM (mm2) from each group was log transformed to normalize data acquired from various lumen sizes and the least square means of each group were calculated. Airways from HA-PBS animals (N= 59) tended to have the lowest ASM, and ASM progressively increased in RA-PBS (N=41), HA-Ova (N=58) and RA-Ova groups (N=38 airways) [-6.41, -6.3, -6.26, -6.17]. There was an excellent correlation between ASM and airway resistance; more ASM was associated with greater airway reactivity (Pearson correlation of 0.99).
Conclusions and Potential Impact: Chronic hypoxia may suppress ASM development, as well as suppress the increase in ASM associated with atopic inflammation. Understanding the mechanisms that inhibit ASM growth under conditions of chronic hypoxia may provide insight into new therapies for asthma.
Epidemiological Assessment of GI BioFire Negative Tests at Parkview Health
Authors: Ratcliffe, B.S. Brycen , Rodgers, R.N. Candace , Stienecker, M.D. R. Scott
Published: October 2019

Background and Hypothesis: With advances in multiplex PCR testing, many gastrointestinal pathogens can be identified in about an hour and provide 23% increased yield of pathogens (Beatty et al., 2016). At our facility, 60.4% of these samples return with no microorganisms found. This study implemented diagnostic stewardship principles by evaluating the negative GI BioFire multiplex results and ordering patterns of providers at Parkview Health.
Design: This retrospective chart review of 300 negative GI BioFire results from 2018 pulled demographics and records from Parkview LIS and EPIC software. Inclusion Criteria: negative gastrointestinal BioFire results. Exclusion Criteria: positive results, negative results outside the study period, any patient data not admitted at Parkview Health.
Results: 57.0% of patients had diarrhea-associated co-morbidities. 14.3% had redundant tests. 51.0% were administered laxatives, 72.3% were administered antibiotics. 73.4% did not follow ACG guidelines stating diarrhea must have persisted longer than seven days. 79.3% had laxatives concurrently administered or did not follow ACG guidelines. 19.7% had endoscopic procedures within eight weeks of assay testing. 14.4% of providers documented a non-C Diff suspicion when ordering assay. 18.9% had non-reported diarrhea consistency. 45.9% were hospitalists, 5.5% were surgeons, and 17.9% were infectious disease or gastroenterologists. 78.3% had at least one C-Diff diarrhea risk factor.
Conclusion and Potential Impact: This data suggests using appropriate algorithms for GI BioFire and utilizing C-Diff antigen testing is more cost effective and should be used before GI BioFire for those with risk factors. Following ACG guidelines, and deferring those administered laxatives would remove over three-quarters of negative panels. Documentation of stool consistency, volume and suspicious etiologies are essential to furthering diagnostic stewardship. These recommendations will save approximately $1,036,399 after consideration for C-Diff antigen testing replacement per year for Parkview Health.
Retinal and choroidal vascular abnormalities contribute to the conversion from dry to wet age-related macular degeneration: a theoretical approach
Authors: Rowe Lucas , Harris Alon , Ciulla Thomas , Chiaravialli Greta , Verticchio Vercellin Alice Chandra , Siesky Brent , Guidoboni Giovanna
Published: October 2019

Background and Hypothesis: Age-related macular degeneration (AMD) is the leading cause of adult blindness in the developed world, and can be classified as one of two types: dry or wet. Abnormalities in retinal and choroidal vasculature may influence dry-to-wet conversion. This study represents a first attempt to use mathematical modeling to characterize the impact of retinal and choroidal blood flow on the oxygenation of retinal layers at various distances from the macula, in healthy individuals and AMD patients.
Experimental Design or Project Methods: The macula is modeled as 7 layers: ganglion cell layer (GCL), inner plexiform layer (IPL), inner nuclear layer (INL), outer plexiform layer (OPL), outer nuclear layer (ONL), photoreceptors layer (PH), retinal pigmented epithelium (RPE). Oxygen supply is provided by the vitreous, the choroid, and by three retinal capillary plexi. Oxygen profiles through the macular tissue are calculated by simulating the balance between O2 supply, consumption and diffusion in: physiological baseline conditions; AMD conditions.
Results: Choroidal vasculature impairment affects tissue more proximal to the macular center, retinal blood flow impairment affects tissue more proximal to the macular periphery, and oxygenation of the foveal avascular zone is not affected by retinal vasculature impairment. The decrease in oxygenation due to retinal and choroidal blood flow impairment in AMD is more prominent in the RPE, PH and ONL in all three anatomical zones of the macula.
Conclusion and Potential Impact: Our mathematical model revealed that reduced choroidal and retinal oxygenation in AMD patients mostly affects the RPE and PH layers, regardless of the distance from the macula. This finding may explain hypoxia inducible factor-1 (HIF-1) production in these layers, which leads to enhanced vascular endothelial growth factor (VEGF) production, causing neovascularization and conversion to wet AMD. Our model suggests that treatment modalities aimed at maintaining stable oxygenation in dry AMD patients may prevent conversion to wet AMD, and reduce vision loss in these patients.
Chamas for Change: Redevelopment of Community Health Volunteer Curriculum for a Group-Based Maternal-Child Peer Support and Microfinance Care Model in Western Kenya
Authors: Rushton, BS Grace , Songok, MMBchB Julia Jerono , Jumah Anjellah , Ikemeri, BSc Justus E. , Ruhl, MD MPH Laura
Published: October 2019

Background: Maternal mortality continues to be the leading cause of death for women of child-bearing age in Kenya. Chamas for Change builds on longstanding cultural practices as a community-based peer support model for pregnant and breast-feeding mothers. Through medical and social education, microfinancing for economic empowerment, and a culture of community accountability, Chamas enables women to advocate for their health and improve health outcomes for their children. With significant success over the past eight years and a recently completed cluster randomized control trial (RCT), the Chamas for Change team is looking to expand the scope of Chamas to additional counties in Kenya and beyond.
Project: To prepare for this transition to scale, the Chamas flipchart curriculum was redeveloped to include additional education topics and restructured to make each of the three cycles (or years) more cohesive and structured. To investigate new topics, brainstorming was conducted within the Maternal, Newborn, and Child Health (MNCH) team. Community Health Volunteers (CHVs) and implementation leads for the program were surveyed and a previously completed focus group discussion with Chamas members was analyzed. Lastly, additional topics were gathered from existing child development and adolescent reproductive health curricula through AMPATH.
Results: Content for new medical and social topics was developed and integrated with existing topics. A total of 24 medical and 24 social topics were established for each of the two initial cycles of the program. Each session includes a reflection, time for socializing and activities, health and social education content, and homework to prompt further reflection.
Conclusions and Impact: This refined flipchart will help to address demands from Chama mothers who are seeking a more robust education, while also further standardizing and broadening the curriculum for CHVs. It will additionally
Tranexamic Acid Increases Early Postoperative Pain and Decreases Time to First Opioid Following Total Knee Arthroplasty
Authors: Seetharam Sachin , Keller Sydney , Ziemba-Davis Mary , Meneghini MD R. Michael
Published: October 2019

Background and Hypothesis: Tranexamic acid (TXA) decreases blood loss in total knee arthroplasty (TKA). However, TXA evoked pain in rats by inhibiting GABA and glycine receptors in the spinal dorsal horn, and caused cellular death in ex vivo and in vitro human periarticular tissues exposed to clinical concentrations of TXA. We evaluated inpatient postoperative pain and blood loss in TKA performed with and without TXA.
Project Methods: 105 consecutive cemented TKAs without TXA were compared to 72 consecutive cemented TKAs with TXA. Procedures were performed by a single surgeon using identical perioperative medical and pain-control protocols. Outcomes included: average of q2-4 hour pain scores during the first 24 hours after PACU discharge, average pain during remainder of stay, final pain score prior to discharge, time in minutes to first opioid after PACU discharge, total opioids in morphine equivalents (MEQs) during the first 24 hours after PACU discharge, average MEQs per remaining days of stay, and mean g/dL pre- to postoperative decrease in hemoglobin. Multivariate analyses accounted for 15 demographics and covariates.
Results: The sex (p=0.393), age (p=0.784), and BMI (p=0.930) of the two cohorts were similar. Mean pain during the first 24 hours was greater (4.1 vs. 3.2, p=0.001), MEQs consumed during the first 24 hours were greater (45 vs. 37, p=0.069), and time to first opioid medication was shorter (326 vs. 414, p=0.023) in patients who received TXA. The decrease in hemoglobin was less in patients who received TXA (-2.2 vs. -2.7, p<0.001).  
Conclusion and Potential Impact: Our hypothesis based on animal and laboratory studies that TXA may increase early postoperative pain was confirmed by three metrics. Consistent with the effective life of TXA, pain and opioid consumption after 24 hours did not differ based on TXA use. Further work is warranted to investigate the nature consequences associated with TXA, relative to its demonstrated benefits for blood conservation.
Telepresence And Provider Communication Predict Satisfaction With Telestroke
Authors: Selch Griffin Tyler , Lyerly Michael J. , Martin Holly , Graham Glenn , Martini Sharyl , Anderson Jane , Damush Teresa , LaPradd Michelle , Ofner Susan , Williams Linda
Published: October 2019

Background: In 2016 the Veterans Health Administration implemented the first nationwide Telestroke program; 800 consults were completed in the first 18 months. Preliminary analysis showed Veterans reported high satisfaction and acceptance of the program. This study sought to understand patient, provider, and hospital-level factors associated with patient satisfaction.
Methods: Patients who received a Telestroke consultation were eligible for a phone interview two weeks later, including standard questions about technology quality, telepresence (how much the encounter felt like face-to-face), Telestroke provider communication, and overall satisfaction. Satisfaction scores ranged from 1-7, (higher = more satisfied), and for analyses were dichotomized as 6-7 indicating high satisfaction vs. < 6. Patient variables including stroke severity (NIH Stroke Scale) were obtained from study records. We used Student’s t-tests and Chi-square tests to compare variables related to patient-reported satisfaction, and used a logistic regression model to determine factors independently associated with high satisfaction.
Results: Over 18 months, 208 interviews were completed and 156 (75%) reported high satisfaction with Telestroke. Patients with more severe stroke were less likely to recall the consultation (p = 0.01). Factors significantly associated with patient satisfaction were higher ratings of the technology (p < 0.0001), telepresence (p < 0.0001), provider communication ratings (p < 0.0001) and overall VA satisfaction (p = 0.01). Among 13 providers with at least 10 consultations, there was no difference in mean patient satisfaction scores. In the multivariate model, telepresence (OR 3.10, 95% CI 1.81-5.31) and provider communication scores (OR 2.37, 95% CI 1.20-4.68) were independently associated with satisfaction.
Conclusion and Potential Impact: Provider qualities, including telepresence and provider ratings, were associated with overall Veteran satisfaction with Telestroke. Technology quality may be necessary but not sufficient to impact patient experience. Training providers to improve telepresence and communication skills could improve patient experience with Telestroke consultation.
The efficacy of P.O.P. on Teen violence and the anti-violence programing in Northwest Indiana and greater Chicagoland area
Authors: Sloss Isaiah , Anokwute Chiamara , Rutland, MD Reuban , McGee, MD Michael
Published: October 2019

Background: Fights, bullying, and gun violence in the Northwest Indiana and Chicagoland middle schools and high schools can have long lasting effects on a student’s physical, emotional, or mental wellbeing. Project Outreach and Prevention on Youth Violence (henceforth POP) is a foundation started by two NWI doctors, Dr. Michael McGee and Dr. Reuban Rutland, looking to do their part in preventing victims of youth violence from reaching their emergency departments. POP also reaches students underrepresented in medicine to breach that gap. Programs such as POP’s Health Professions Enrichment Program (HPEP). POP also utilizes school rallies and violence prevention seminars throughout the year to reach at risk youth. Activities at these rallies include Stop the Bleed training, where students are trained and certified by professionals on how to stop a bleed to various parts of the body. POP also utilizes local celebrities, artists, public figures, and community leaders to incentivize the participation of their target population. Does implementation of anti-violence events with school age students by professionals and community leaders decrease incidences of violence or increase interest in college/STEM professions?
Hypothesis:

 There is an increase in the interest in college/STEM professions.
 There is a decrease in incidences of violence at schools with active SAVE chapters or participating in an anti-violence event

Experimental Design or Project Methods: Surveys, Observations, Student Interviews, Intervention, Cohort Study, Program evaluations, Longitudinal Study (Go to each school nearby and request the info) The study’s population includes students from 7th grade through high school seniors located throughout 13 schools in the northwest Indiana region. POP
Results: Analyze Survey Data and interviews for positive correlation with the data. 
Conclusion/Potential Impact: Pending Review; The potential impact this project yielding positive results would help demonstrate efficacy of POP and enable us to seek more funding opportunities and implement these programs on a wider scale and hopefully being able to see lower instances of violence in areas where programing takes place.
Gpr17 signaling decreases GLP-1 secretion in GLUTag cells
Authors: Stalbaum Katherine , Yan Shijun , Ren Hongxia
Published: October 2019

Background and Hypothesis: The incidence of obesity and diabetes continues to rise in devastatingly high proportions, making the need for safe, affordable, and effective treatment increasingly apparent. We discovered that the orphan G protein-coupled receptor 17 (Gpr17) is expressed in endocrine cells in the brain and gut and may have an important role in metabolic regulation. Glucagon-like peptide 1 (GLP-1), an incretin hormone secreted from enteroendocrine cells, is a strong insulin secretagogue and suppresses appetite. We hypothesized that Gpr17 signaling decreases GLP-1 secretion in gut enteroendocrine cells.
Experimental Design or Project Methods: In order to investigate the role of Gpr17 in GLP-1 secretion, we measured GLP-1 secretion in a murine enteroendocrine cell line (GLUTag cells) that expresses Gpr17 and the proglucagon gene and secretes GLP-1 in a regulated manner. GLUTag cells were stimulated with glucose or lipid, oleoyl-lysophosphatidylcholine (LPC), in the presence or absence of MDL29,951, a synthetic Gpr17 agonist. After a 2-hour incubation, we measured GLP-1 in the media and cell lysates to determine the percentage of secreted GLP-1.
Results: Cells treated with glucose and MDL29,951 had decreased GLP-1 secretion compared to glucose alone, however, the difference was not significant. Cells treated with LPC and MDL29,951 had a significant decrease in GLP-1 secretion compared to LPC alone.
Conclusion and Potential Impact: Gpr17 activation by MDL29,951 decreased GLP-1 secretion in GLUTag cells stimulated by both glucose and lipid, which supports our hypothesis that Gpr17 signaling regulates GLP-1 secretion. Therefore, Gpr17 may be a potential pharmacological target for combating obesity and diabetes.
Inhibition of P65 and PDK2 Increases Cytotoxicity and Radiation Sensitivity of Pancreatic Cancer Cells
Authors: Streveler Joshua A. , Chin-Sinex Helen , Mendonca Marc S.
Published: October 2019

Background: Pancreatic ductal adenocarcinoma (PDAC) is currently the 4th leading cause of cancer death in the US because of late detection and resistance to chemotherapy and radiation. This tumor resistance is partially due to high activity of the transcription factor, NF-κB which promotes cell survival and pyruvate dehydrogenase kinase 2 (PDK2) which regulates aerobic glycolysis (Warburg Metabolism), cell proliferation, and inhibition of apoptosis. We have shown that simultaneous treatment with the chemical NF-κB inhibitor DMAPT and the Warburg inhibitor DCA is cytotoxic and enhances radiation-induced cell death in human PDAC cells. We have also shown that double gene knockdown of the P65 (RELA) subunit of NF-κB and PDK2 by siRNA increased cytotoxicity and radiation sensitivity of pancreatic cancer cells but to a lesser extent than DMAPT and DCA treatment.
Experimental Design: Western blots suggested the siRNA did not completely suppress p65 and PDKS protein levels. To test the hypothesis that more complete inhibition of NF-κB and PDK2 is required to increase cytotoxicity in PDAC cells, we transfected the Mia PaCa-2 cell line with appropriate siRNA and exposed the cells to the chemical inhibitors DMAPT and DCA. The cells were plated and followed by irradiation with either 0, 2, 4, or 6 Gy of 160 kVp X-rays
Results: Compared to previous experiments that utilized siRNA knockdown alone, the combination of dual drugs and dual siRNA shows an increase in both cytotoxicity and radiation-induced cell killing (p< 0.001, t test)
Conclusion: The data suggest that a more definitive approach such as utilizing CRISPR-Cas9 may be required to ensure complete gene knockdown of p65 and PDK2 to elucidate the mechanism(s) by which simultaneous inhibition of NF-kB and PDK2 inhibition enhance cytotoxicity and radiation-induced cell killing in pancreatic cancer.
Type 2 Diabetes-Driven Alterations in Bone Healing and Angiogenesis
Authors: Sun Seungyup , Bhatti Fazal Ur Rehman , Dadwal Ushashi C. , Babu Deepa Sheik Pran , Perugini III Anthony J. , Awosanya Olatundun D. , Blosser Rachel J. , Tersey Sarah A. , Orr Kara S. , Randhave Karishma R. , Li Jiliang , Yoder Mervin C. , Evans-Molina Carmella , Kacena Melissa A.
Published: October 2019

Background and Hypothesis: Type 2 diabetes (T2D) is prevalent in the United States. T2D patients are at risk for impaired fracture healing due to decreased angiogenesis, which is required for successful bone regeneration. Bone morphogenetic protein-2 (BMP-2) is often used to help orthopedic surgeons with bone healing in difficult cases. Here, we begin characterizing the mechanism by which T2D alters bone healing with or without BMP-2 treatment. We hypothesize that T2D impairs fracture healing by decreasing angiogenesis and endothelial cell function.
Project Methods: Using Tie2-CreER;Td-Tomato mice (Tie2CreERT+), we established a high fat diet (HFD)-induced T2D mouse model to compare with control low fat diet (LFD)-fed mice. Mice underwent testing to confirm the T2D-like metabolic phenotype, underwent a femoral critical-size defect surgery that was treated with either saline or BMP-2, and were then assessed biweekly by X-ray imaging over the course of 12 weeks. Finally, bone marrow-derived endothelial cells were collected from these mice to assess changes in endothelial colony and tube formation in vitro.
Results: Results showed that the HFD mice acquired the T2D metabolic phenotype. Fracture healing was impaired in the HFD mice, even with BMP-2 treatment. The isolation of BMECs was confirmed by visualization of fluorescent Tie2+ cells. Unexpectedly, in vitro tube formation assays indicated that HFD improved vessel-like formation properties. BMP-2 treatment appeared to improve some vessel-like formation properties compared to control treatment.
Conclusion and Potential Impact: This study is ongoing. Further data will need to be collected to better characterize differences in bone healing and angiogenesis in the healing femurs. Still, these data reveal the mechanisms by which T2D impairs bone healing and demonstrate the important difference between examining endothelial cells in vitro vs. in vivo. Future investigations will examine if thrombopoietin, which our group has previously shown to improve both fracture healing and angiogenesis, may be a more effective treatment than BMP-2 in this model.
Identification of gene expression alterations in C. elegans adr-2 mutants using high- throughput sequencing
Authors: Townsend Jackson , Hundley Heather A.
Published: October 2019

Background and Hypothesis: RNA editing is one of several mechanisms regulating gene expression. One type of RNA editing, the deamination of adenosine to inosine, is carried out by ADAR enzymes. ADAR enzymes are essential for neural function and aberrant editing is implicated in various forms of neuropathology. C. elegans lacking the RNA editing enzyme, ADR-2, are viable allowing us to ascertain how loss of RNA editing affects neural gene expression. The effects of loss of adr-2 on neural gene expression will be analyzed in both the first larval (L1) and young adult stages. We hypothesize that the transcriptome will change depending on life stage and the presence of ADR-2.
Methods: Three replicates of neural cells isolated from wild type and adr-2(-) L1 and young adult stage animals were obtained. Total RNA was extracted from each population and mRNA was isolated using an oligo-dT bead. The mRNA was fragmented, and reverse transcribed to generate a complentary DNA (cDNA) library. The cDNA was sequenced by a facility at Indiana University. Quality of the library was evaluated using FASTqc. DE-seq2 software evaluated the differential gene expression.
Results: I examined differential gene expression in two life stages of the WT and adr-2 neural samples. After obtaining the differentially expressed genes, the portions of the transcriptome that require ADR-2 was determined. WT young adults showed increased (3715) and decreased (2504) expression of neural genes when compared to the L1 stage. Many differentially expressed genes required adr-2 (~40% of the upregulated and 78% of the downregulated genes.) In addition, some genes were uniquely altered (631 upregulated, 196 downregulated) in the absence of adr-2.
Conclusion and Potential Impact: The life stage and presence of ADR-2 alter the neural transcriptome and this function changes throughout development. Future studies will determine whether these genes are altered due to the lack of RNA editing or binding by ADR-2.
Development of a High-Fidelity, 3D Printed Otoplasty Surgical Simulator
Authors: Wallace, M.S. Chelsey , Nourmohammadi, Ph.D. Zahra , Zopf, M.D., M.S. David A.
Published: October 2019

Background and Hypothesis: Protruding ears, also known as prominauris, are prevalent worldwide at an occurrence rate of about 5%. Children with prominauris report lower self-esteem and experience increased teasing and social isolation at school. From a functional standpoint, protruding ears can make wearing prescription glasses difficult. This increased stress and anxiety and impaired functionality lead children and families to seek treatment. One of the most effective treatments for protruding ears is otoplasty. This procedure involves an incision in the back of the ear and the placement of non-resorbable sutures to reform the ear. Alternatively, the procedure can be performed using an incisionless technique. As this is an elective procedure done commonly in children, adequate education of medical trainees is critical to ensure the proper level of skill is attained and patient satisfaction is maximized.
Currently, teaching otoplasty is done with cadavers and supervised procedures with an attending. Surgical simulators are employed in the instruction of a variety of surgical procedures and allow residents to practice in a zero-risk environment. In addition, 3D printing has facilitated the development of surgical simulators allowing for a more cost-effective, consistent, and anatomically correct simulator. We developed an ear model made from silicone for trainees to practice traditional and incisionless otoplasty.
Project Methods: The otoplasty surgical simulator was developed by isolating an ear from a computed-tomography scan in the Materialise software to create a 3D model. This model was then altered to create a negative mold. The mold was printed using fusion deposition printing with 1.75 MM polylactic acid filament. After printing, the mold was filled with Dragon Skin Silicone Shore 20 to simulate ear cartilage. The model was then coated in a layer of Dragon Skin Silicone Shore 10 to simulate a layer of skin.
Conclusion and Potential Impact: This otoplasty simulator will next be validated by expert surgeons and then used in a surgical simulation workshop for surgical trainees. Because of the low-cost of the surgical simulator and the ease of manufacturing, this simulator can also be used to train surgeons abroad where access to surgical training may not be readily available.
A novel ferrochelatase inhibitor as a therapy for ocular neovascularization
Authors: Waller Sydney , Pasha Sheik Pran Babu Sardar , Lambert-Cheatham Nathan , Corson Timothy W.
Published: October 2019

Background and Hypothesis: Ocular neovascularization, characterized by the abnormal growth of new blood vessels, underlies blindness-inducing diseases such as wet age-related macular degeneration (AMD) and proliferative diabetic retinopathy (PDR).
This neovascularization can be either choroidal (in the case of wet AMD) or retinal (in the case of PDR). Inhibition of vascular endothelial growth factor is the target of current FDA approved therapies, however the need for intravitreal injections poses a drawback to such treatments. Furthermore, many patients are non-responders or refractory. Given this, the discovery of novel therapeutic targets is warranted.
Ferrochelatase (FECH) is an enzyme involved in heme synthesis. The Corson lab has previously shown FECH to be necessary for angiogenesis both in vitro and in vivo. The Corson lab has been successful in developing novel compounds that inhibit FECH in vitro, based on high-throughput screening hits. The objective of this project was to test one such novel FECH inhibitor in a choroidal neovascularization mouse model.
Experimental Design: FECH inhibition was investigated in the laser-induced choroidal neovascularization model. This model involved creating a laser-induced lesion into the retinal pigment epithelium/choroid of mice aged 6-8 weeks; this promotes neovascularization. The FECH inhibitor compound was delivered once intravitreally at the time of the laser. Neovascularization was analyzed in vivo using optical coherence tomography and fluorescein angiography one and two weeks after laser, then quantified ex vivo by isolectin staining.
Results: There was a trend toward decreased ocular neovascularization with the administration of a FECH inhibitor, as measured by optical coherence tomography seven days after laser treatment. However, further analyses are ongoing to validate this finding.
Conclusion and Potential Impact: The administration of a FECH inhibitor compound intravitreally may result in a decrease in CNV. This may be useful as it could lead to FECH serving as a target for future therapies.
Role of CaMKK2 in Mechanically Induced Bone Formation
Authors: Warrick Adam J. , Sankar Uma
Published: October 2019

Background and Hypothesis: Mechanical stimulation of bone results in the translation of external forces into a cascade of structural and biochemical changes which work to increase bone density and decrease fracture healing time. The specific mechanisms contributing to these processes are areas of active investigation. Ca2+/calmodulin-dependent protein kinase kinase 2 (CaMKK2) is a serine-threonine protein kinase with key roles in both the anabolic and catabolic pathways of bone remodeling. We hypothesize that the absence of CaMKK2 potentiates an increase in bone density as a response to mechanical stimulation.
Experimental Design or Project Methods: The right ulna of anesthetized C57BL/6 mice were loaded for 220 cycles at 2 Hz and with peak forces specific to both sex and genotype. Loading was completed using an electro actuator (Bose ElectroForce 3200; EnduraTEC, Minnetonka, MN, USA) and was repeated on days 3, 5, 8 and 10 after the initial procedure. The non-loaded left ulna served as an internal control. Calcein and alizarin red were administered intraperitoneally on days 9 and 16 respectively. Mice were sacrificed on day 19 after the initial load; blood and long bones of the lower limbs were collected for analysis.
Results: Bone volumetric analyses will be measured using microcomputed tomography, bone formation rate will be assessed using dynamic histomorphometry measurements of double fluorochrome labeling, and cellular and molecular mechanisms will be assessed using histology, immunohistochemistry and real-time reverse transcription-polymerase chain reaction. These data are currently forthcoming.
Conclusion and Potential Impact: Clinical outcomes of conditions ranging from stress fractures to osteoporosis may be improved by an increased understanding of the mechanisms through which bone growth is augmented. Expanded knowledge of these pathways may provide opportunities for the development of novel therapies which decrease healing times in the event of injury and increase bone density to combat degenerative disease states.
Measuring Subjective Cognitive Decline in Older Adults: Harmonization between the Cognitive Change Index and the Measurement of Everyday Cognition Instruments
Authors: Wells Lindsey F. , Risacher Shannon L. , Saykin Andrew J.
Published: October 2019

Background and Hypothesis: Self and informant reports of everyday cognitive functioning have been shown to be associated with incipient neurodegenerative disease. The 20-item Cognitive Change Index (CCI) [1] and the 39-item Measurement of Everyday Cognition (ECog) [2] were each developed to characterize early changes in cognitive function. The aim of this study is to examine the relationship between the CCI and ECog self- and informant-evaluations to improve early detection and longitudinal assessment of cognitive decline for observational research and clinical trials.
Experimental Design or Project Methods: Self-evaluation data included 803 participants (49.3% male, mean age=69.9yrs) from the Alzheimer’s Disease Neuroimaging Initiative (ADNI), the Indiana Alzheimer Disease Center (IADC), and the Indiana Memory and Aging Study (IMAS). Informant-evaluation data included 288 participants (39.9% male, mean age=66.9yrs) from the IMAS and IADC studies. Study participants from 2012 to 2019 were included if they had completed both the CCI and ECog at the time of their baseline visit. Pearson correlations, regression analyses, and frequency distributions were used to assess the relationship between the CCI and ECog. Sex, years of education, race, ethnicity, APOE ε4 carrier status, and baseline diagnosis were also analyzed as potential relevant covariates.
Results: CCI and ECog total scores were highly correlated for the self (r=0.778, p<0.001) and informant (r=0.892, p<0.001). The highest associations were between the mean item scores for each instrument for both self (r=0.801, p<0.001) and informant (r=0.913, p<0.001). Frequency distributions showed distinct patterns between self and informant total scores. Polynomial regressions for self (R2=0.673) and informant (R2=0.873) total scores were used to create a translation table between the CCI and ECog.
Conclusion and Potential Impact: Self and informant total scores can be harmonized and translated between the CCI and ECog to facilitate cross-sectional and longitudinal assessment of perceived cognitive change, an important patient reported outcome
[1] Rattanabannakit et al. (2016). Journal of Alzheimer’s Disease, 51(4), 1145-1155.
[2] Farias et al. (2008). Neuropsychology, 22(4), 531-544.
Amyloid and Tau in Neurodegeneration
Authors: Williams Acacia , Hallinan, PhD Grace , Gomez Ponce, MS Maria Teresa , Vidal, PhD Ruben
Published: October 2019

Background and Hypothesis:Familial British dementia (FBD) and familial Danish dementia (FDD) are two autosomal dominant neurodegenerative diseases caused by mutations in the BRI2 gene. FBD and FDD are characterized by widespread cerebral amyloid angiopathy (CAA), parenchymal amyloid deposition, and neurofibrillary tangles.Previous studies have shown that accumulation of Danish amyloid peptide (ADan) induces hyperphosphorylation of tau and cell cytotoxicity; however, progress in our understanding of the interaction between ADan and tau has been hindered by the lack of antibodies against ADan. In addition, whether the British amyloid peptide (ABri) has a similar effect on tau phosphorylation remains unknown. The main goals of our project are to generate monoclonal antibodies against ADan and to test whether the ABri peptide induces hyperphosphorylation of tau and cell cytotoxicity.
Experimental Design or Project Methods:A peptide homologous to the C-terminus of the ADan peptide was used to immunize 5 mice. Serum samples were tested from mice with high ELISA titers and the best 2 animals were used for cell fusion. Cell culture supernatants were screened by ELISA, western blot and immunohistochemistry. ABri peptides were used as negative controls.HEK cells expressing human tau were used to assess the interaction between ABri and tau. We performed immunocytochemistry and Western blot analysis to investigate tau phosphorylation.
Results:A total of 11 clones tested positive by western blot. 3 clones tested positive for ABri and were discarded. 5 clones were tested by immunohistochemistry. 3 of the positive clones will be used for subcloning to complete clonality. Work in progress on the interaction between ABri and tau will determine whether ABri induces tau hyperphosphorylation in cell culture.
Conclusion and Potential Impact:Our work will provide novel tools to study the interaction between amyloid and tau in neurodegenerative diseases and may uncover possible new targets for pharmaceutical intervention in tauopathies.
The Effect of Biofilm-Forming Bacteria on Inflammasome Activity
Authors: Williams, BS Georgia Lea , Santra, PhD Suman , Roy, PhD Sashwati
Published: October 2019

Background and Hypothesis: Over 75% of chronic wounds have biofilm infection. Biofilm infection derails the overall process of wound healing. Biofilm bacteria have developed strategies to subvert macrophage inflammatory response. The underlying mechanisms of such subversion are currently unknown. Inflammasomes are multiprotein, intracellular complexes that contain caspase-1. Caspase-1 is responsible for cleavage and activation of cytokines IL-1β, a major pro-inflammatory cytokine in chronic wounds. We hypothesized that biofilm bacteria subvert macrophage inflammatory response by attenuating caspase-1 activity and thereby reducing IL-1β. The primary objective of the study was to determine Caspase-1 activity and IL-1β production by blood monocytes derived from patient blood samples exposed to biofilm infection ex vivo and clinical wound macrophages.
Methods: Wound macrophages were isolated from chronic patients seen at Indiana University Health Comprehensive Wound Center (CWC). Blood monocyte derived macrophages (BMDMs) were exposed to conditioning media from isogenic mutant strains SA300ΔsarA or SA300ΔrexB (derived from Staphylococcus aureus USA300LAC) as hypo- and hyper-biofilm forming mutants. Wound macrophages were isolated from wound fluid with known bacterial content. The IL-β release and caspase-1 activity from macrophages were determined using ELISA, PCR, and a colorimetric assay.
Results: Both RT-PCR and ELISA independently exhibited a significant reduction in IL-1β production in macrophages exposed to conditioned media from hyper-biofilm forming SA300ΔrexB as compared to hypo-biofilm forming SA300ΔsarA mutant.The Caspase-1 activity was significantly reduced in the macrophages challenged with hyper-biofilm forming bacteria as compared to hypo-biofilm forming mutant.
Conclusion and Potential Impact: Biofilm forming bacteria leads to an attenuation in production of IL-1β from macrophages indicating a subversion of inflammatory response. The caspase-1 activity data exhibited a significant role of caspase-1 in biofilm mediated subversion of the inflammatory response indicating a reduction in inflammasome activity. A clear understanding of the mechanisms of biofilm mediated suppression of host response will help improve therapeutic strategies against wound biofilm infection.
Predictors of Quality of Life after Liver Transplant
Authors: Wu Joey , Desai, MD Archita
Published: October 2019

Background and Hypothesis: The impact of chronic liver diseases on patients and their family member is often understated and understudied. Chronic liver diseases can sometimes progress to a need for Liver transplant (LT). While recent studies have described quality of life (QOL) at different stages of liver disease, the impact of the patient’s QOL in LT survivors has not been examined. The importance of studying QOL in patients is due to its effect on the survivorship of LT recipients. We hypothesize that QOL in LT patients is lower than the general population. Our aim was to describe predictors of QOL in a well-described cohort of LT patients.
Methods: Patients were enrolled at the Digestive and Liver Disease Liver clinic at Indiana University Hospital. All patients over the age of 18 were approached, if patients consented to the study, they were then enrolled during their liver follow up visit. The PROMIS survey was administered on an iPad and completed during the clinic visit. Survey were then scored and analyzed.
Results: The T-scores for post liver transplant patients are lower in physical function, anxiety and depression, but higher in general life satisfaction compared to the general population. LT recipients have similar T-scores in Fatigue, Sleep disturbance, ability to participate in social activities, and pain interference compared to the general population.
Conclusion and Potential Impact: Previous diagnosis of PBC, HCC, diagnosis of depression, household income, insurance status, Charlson Comorbid Index and number of non-transplant related medications have the highest association with quality of life. Further enrollment is needed to increase the power of the study. However, this can inform physicians the importance to taking these factors in to consideration in order to improve the QOL in LT recipients.
Urban-Rural Differences in Indiana Pediatric Firearm Trauma
Authors: Wuerch Cory , Raymond, MPH, CSTR, CAISS Jodi , O'Neil, MD, MPF, FAAP Joseph , Landman, MD, MPH, FAAP, FACS Matthew P.
Published: October 2019

Background and Hypothesis: Several studies have evaluated differences in firearm injuries among children and adolescents based on population. However, many of these studies exclude patients who die before arriving at a trauma center. We therefore hypothesize that important population-based differences in pediatric firearm injuries may be uncovered with inclusion of both pre-hospital firearm mortalities and patients treated at a tertiary children’s hospital.
Methods: Patients less than 15 years of age who sustained a firearms-related injury/death between the years 2012 and 2018 were identified in: (1) death certificates from the Office of Vital Statistics State of Indiana and (2) Riley Hospital for Children Trauma Registry. Counties were classified as either urban, midsized, or rural based on the National Center for Health Statistic’s population data. Bivariate analyses were used to analyze important variables with statistical significance set at p<0.05.
Results: A total of 222 patients were identified (77% male, mean age = 11.3 ± 4.2 years). The median age of firearm injury survivors was 13 (IQR 7-14), while the median age of nonsurvivors was 14 (IQR 11-15). The county population was associated with injury intent, where the frequency of assaults/homicides was higher than expected in urban counties compared to midsized and rural counties (p<0.05 and p<0.001, respectively). Suicide frequency was higher than expected in rural counties (p<0.001). Race was associated with injury intent (p<0.001). Rural and midsized counties had higher than expected mortalities compared to urban counties (p<0.001 and p<0.05, respectively). Attempted suicides in rural areas were more likely to be associated with mortality than attempts in urban areas (p<0.001).
Conclusion and Potential Impact: Important differences exist between firearm injuries based on where they occur. The findings presented here will inform public health initiatives aimed at reducing firearm injury and death in Indiana.