bluuebunny/crossref_metadata_2025 huggingface dataset:
Association Between Peripheral Eosinophil Counts and Infant Lung Function
Authors: Close BS Alexander , Slaven MS James , Ross MS Sydney , Kloepfer MD MS Kirsten
Published: October 2019

Background and Hypothesis: Some children with allergic asthma have peripheral eosinophilia early in life, suggesting that physiological changes in the lung begin during infancy. While this association has been established, studies have not examined if a link exists between eosinophilia and physiological airway measurements during the first year of life. It is hypothesized that systemic eosinophilia during the first year of life is associated with lower infant pulmonary function measurements.
Experimental Design or Project Methods: A birth cohort study investigating the development of the upper airway microbiota over the first year of life was utilized for this study. Systemic absolute eosinophil counts were examined to determine if eosinophilia is associated with decreased infant pulmonary function test (iPFT)values. Blood and iPFT data were collected at 3 and 12 months.
Results: 28 participants had blood samples collected. 15 samples were taken at 3 months and 13 at one year. 16 subjects were male, with a majority identifying as African American (14). The average height and weight were 68.0 cm and 8.6 kg. The median absolute eosinophil count was 0.2010^3/uL. Infant iPFT data included tidal volume, respiratory rate, expiration time, tPTEF/tE, resistance, and compliance. No significant correlations were identified between eosinophil counts and iPFT data. This held true when all time points were combined and when examining each visit separately. The strongest correlation coefficient was -0.36 between absolute eosinophil count and FEV0.5. The estimated number of samples needed to power a study examining the association between FEV0.5 and eosinophil counts is 80 samples.
Conclusion and Potential Impact: Based on preliminary results, there are not enough samples to achieve significant correlations supporting the original hypothesis. The next step is to analyze more blood from current study participants. It is also planned to examine the association between the airway microbiome and eosinophilia to determine if early allergic markers in the blood may results from airway dysbiosis.
Body Morphometrics in NAFLD Population
Authors: Dhami Harkeerat , Samala Niharika
Published: October 2019

Introduction: NAFLD is one of the common causes of liver disease in the US and is commonly associated with metabolic syndrome. Among obese, prevalence of NAFLD is 7090%. We wanted to determine body morphometrics in NAFLD.
Methods: All individuals presenting to Indiana University Hospital with NAFLD were approached to participate in cross-sectional study. All participants were offered beverage, diet (REAP) questionnaires and body composition analysis using InBody 570, which utilizes bioelectrical impedance.
Results: Of the 321 NAFLD individuals enrolled, 256 completed body morphometric analysis. Mean age of the cohort was 51.58 ± 13.54, 58% were female, 297 White and had a mean BMI of 35.92. 76% were obese, 48% had type 2 diabetes, 49.2% had hypertension, 38.6% had dyslipidemia, and 20.5% had obstructive sleep apnea. Despite having similar BMI, females had lower lean body mass (51.01 vs 70.51) and skeletal muscle mass (28.05 vs 39.70), higher body fat mass (46.71 vs 41.04) and percent body fat (46.59 vs. 35.7). Regular coffee consumers had lower BMI (35.3 vs 38, p=0.038), but lower body fat mass (39.9 vs 46.2, p=0.01), percent body fat (41.1 vs 44.4, p=0.05) and higher lean body mass % (58.8 vs 55.5, p=0.049). Processed meat consumption was associated with higher BMI (39 vs 35.3, P=0.01), percent body fat (45.5 vs 42, p=0.04), and lower lean body mass percentage (54.5 vs 58.2, P=0.04). Similar trends were seen with consumption of high sodium processed foods and watching television for ≥ 2 hours/day.
Conclusion: Among individuals with NAFLD, we saw a higher female preponderance, who were found to have unfavorable body morphometrics despite similar BMI as males. Consumption of high sodium processed food and meat and excess screen time have unfavorable, while regular coffee drinkers have favorable body morphometrics, which offer modifiable measures for risk factors associated with NAFLD.
Neonatal Follow Up Study: A Retrospective Chart Review of Developmental Outcomes
Authors: Wolfe, MD Heather , Gilham, NP Denise , Stace, RN, CPN Susan , Emili, BS Uchechukwu
Published: October 2019

Background and Hypothesis: According to the CDC, Preterm birth is defined as a baby born before 37 weeks of gestation. 1 In 2016 preterm birth affected 1 in every 10 infant born in the United States.1 Preterm birth has been known to cause many birth defects along with developmental delays and has attributed to 17% of all infant death. Neonatal Follow Up clinics or programs have been instrumental in helping infants catch up developmentally.3 We believe that with the NFC, we will be able to develop a standard for preterm babies and the normalization of their development.
Experimental Design or Project Methods: A retrospective chart review was conducted to determine the developmental outcome of infants that were born prior to 37 weeks’ gestation. Infants were divided into extreme, very, and moderate preterm cohorts. Test of Infant Motor Performance (TIMP), Ages and Stages Questionnaire (ASQ), Bayley Scales of Infant Development (BSID) and Modified Checklist for Autism in Toddlers (M-CHAT) scores were used to determine a standard for when preterm babies in each cohort normalize developmentally.
Results: Analysis of the data showed that the developmental turning point for each cohort varies. Within the extreme preterm cohort, one can see that there was no clear turning point for development. For the very preterm group, by TIMP 2, one could see a turning point and a decrease in delays. Lastly, moderately preterm cohort, the number of delays was seen tot decrease from the administration of TIMP 2 and continue in this trend.
Conclusion and Potential Impact: Through the administration of these developmental tests, physicians are able to track the development of preterm babies. We concluded that with the proper follow up, one can see an improvement in development as early as the administration of TIMP 2.
1) Preterm Birth https://www.cdc.gov/reproductivehealth/MaternalInfantHealth/PretermBirth.htm 2) Suave, R., & Lee, S. K. (2006). Neonatal follow-up programs and follow-up studies: Historical and current perspectives. Paediatrics and Child Health. http://doi.org/10.1093/pch/11.5.267 3) Voller, S. M. B. (2018). Follow-Up Care for High-Risk Preterm Infants. Pediatric Annals, 47(4). http://doi.org/10.3928/19382359-20180325-03
“Women Never Use Drugs Alone”: Assessing Stigma & Access to Care among Women who Use Drugs.
Authors: Essex Amanda , Lawrence, Ph.D. CFLE CHES Carrie , Turner Brooklyn Sean
Published: October 2019

Background
Substance misuse remains a significant health threat in communities across Indiana. Despite the 2015 HIV outbreak in Scott County, Indiana’s health systems continue to lack the capacity to reduce health harms associated with substance misuse. Unlike general patient populations, people who use drugs (PWUD) face various social and structural barriers that impede access to health care and result in poorer health outcomes. Such impediments are of more significant concern for pregnant women who use drugs (PWWUD) who experience greater stigma, complex health needs, and require more specialized care. The purpose of this study was to assess access to healthcare and related services among women of childbearing age with a history of substance misuse.
Methods
For this qualitative study, participants (n=20) completed a sociodemographic questionnaire and semi-structured interview. Interview questions included perceptions of their overall health, healthcare experiences, and how to improve access to and retention in these services.
Results
The results reported reflect a thematic analysis of the interview transcripts. Two key care barriers identified were: (1) experiences of stigma related to professionals’ attitudes towards PWWUD and (2) fear of losing custody of their child as a result of physician mandated reporting to child welfare.
Conclusions and Recommendations
Addressing social and stigma related barriers experienced by PWWUD are key to increased linkage to and retention in care as well as improved health outcomes. Additionally, our findings call for mandated student and physician education on patients who use drugs as well as reform of mandated reporting laws to reduce barriers and increase care access among PWWUD.
Patellar Tilt in Two Modern Total Knee Arthroplasty Systems: A Matched-Cohort Analysis
Authors: Estes, BS Braeden W. , Pitz, BS Lauren , Deckard, BSE Evan R. , Meneghini, MD R. Michael
Published: October 2019

Background and Hypothesis: Excessive patellar tilt is undesirable in total knee arthroplasty (TKA) and can result in patellofemoral complications and premature failure/wear. Few studies have investigated patellar tilt between TKA systems with different femoral component geometries and bearing articulations. The purpose of this study was to (1) compare patellar tilt between TKA systems with differing femoral component geometries and (2) evaluate related differences in patient-reported outcome measures (PROMS). The study hypothesis was neutral patellar tilt would correlate with improved postoperative PROMS.
Experimental Design or Project Methods: 380 symmetric trochlear groove TKAs were matched to 380 asymmetric lateralized trochlear groove TKAs on age, sex, body mass index, and preoperative patellar tilt. All cases were performed by one surgeon between 12/2010 and 10/2018. Patellar tilt (in degrees) was measured by two independent blinded raters on preoperative and 4-week postoperative merchant view radiographs per the modern Knee Society Radiographic Evaluation System. Prospectively collected and validated PROMS including UCLA Activity Level, components of the Knee Society Score, and Likert satisfaction were evaluated at minimum one-year.
Results: There were no differences between study cohorts for demographics, covariates, or preoperative patellar tilt (p≥0.479). Asymmetric lateralized trochlear groove TKAs had significantly less postoperative patellar tilt compared with symmetric trochlear groove TKAs (p<0.001). Patellar tilt and femoral implant type had no correlation to pain scores (Knee Society), UCLA Activity Level, or satisfaction (p≥0.138); however, postoperative patellar tilt was significantly less for patients who stated their knee “always” felt normal compared to patients who stated their knee “sometimes” felt normal but not “never” felt normal (p=0.033).
Conclusion and Potential Impact: Findings suggest implant type and patellar tilt have minimal effect on overall PROMS. However, minimizing patella maltracking may provide patients with a more normal feeling TKA and could potentially provide long-term benefits of preventing polyethylene wear or other complications leading to premature implant failure.
Comparison of Technical Outcomes in Instrumented Posterolateral Fusion (PLF) with and without Transforaminal Lumbar Interbody Fusion (TLIF) performed using Silicon Nitride cages
Authors: Gray, B.A. Mitchell , Davis, B.S. Kyle , Jagger Lauren , Smith, M.D. Micah
Published: October 2019

Background: Posterior lumbar fusion is performed for lumbar degeneration that leads to spinal stenosis and spondylolisthesis. Two common approaches include PLF and TLIF, with a current lack of consensus as to the superior approach. The objective of this study is to compare fusion rates, spinal parameters and complications for both surgical approaches using the silicon nitride cage (Si3N4). Our hypothesis is TLIF with a Si3N4 cage will have higher fusion rates, improved technical outcomes and fewer complications when compared to PLF alone. The Si3N4 cage has advantageous surface properties compared to other interbody cages, promoting theoretically higher fusion rates for TLIF procedures.
Methods: A retrospective chart review of 102 spinal fusion patients (PLF=17, TLIF=85) was performed. One spine surgeon performed the fusions and reviewed pre-operative and post-operative radiographs. Measurable outcomes included fusion rates, surgical complications and pelvic/spinal radiographic parameters. Radiographic parameters included restoration of lumbar lordosis (LL), segmental lordosis (SL), pelvic incidence (PI), pelvic tilt (PT), disc height (DH) and foraminal height (FH). Patients who had ≥1 year follow up radiographs were included in analysis (PLF=16, TLIF=48).
Results: TLIF patients with a Si3N4 cage had improved fusion rates (PLF=81.8%, TLIF=100% p=0.003), lumbar lordosis (PLF=-4.38o TLIF=3.15o p=0.001), disc height (PLF=0.55mm, TLIF=4.61mm p=<0.001), foraminal height (PLF=-0.05mm, TLIF=2.41mm p=0.036) and a lower incidence of PI-LL mismatch (PLF=46.15%, TLIF=7.5% p=0.004). No statistically significant difference was found for surgical complications (PLF=11.1%, TLIF=17.6%) or segmental lordosis (PLF=-1.00mm, TLIF=1.17mm). An age difference of statistical significance was also found between the two patient populations (PLF=61.9, TLIF=54.1 p=0.018).
Conclusion: Despite the difference in age between the procedure groups, TLIF with a Si3N4 cage proved to be superior in fusion rates, lumbar lordosis, PI-LL mismatch, disc height and foraminal height restoration.
Gastric Per-Oral Endoscopic Myotomy (G-POEM) for the Treatment of Gastroparesis: A Single Center Experience
Authors: Gregor Lennon , Wo John , DeWitt John , Gupta Anita , Yim Brandon , Mendez Martha , Al-Haddad Mohammad
Published: October 2019

Background and Hypothesis: Gastroparesis is a difficult-to-treat disease that has variable presentation. Treatments for gastroparesis include pharmacotherapy, botulinum toxin injection into the pylorus, or surgical pyloroplasty. Based on patient response to pyloric directed therapies, it is theorized that endoscopic pyloromyotomy will prove to be an effective minimally invasive treatment. Early feasibility studies of endoscopic pyloromyotomy (G-POEM) demonstrated safety and early clinical success. Our aim is to assess the safety and sustained clinical efficacy of the G-POEM procedure.
Experimental Design or Project Methods: Patients with confirmed diagnosis of gastroparesis on gastric scintigraphy were assessed using PAGI-SYM to determine symptom severity. EndoFLIP of pylorus and four-hour solid phase gastric emptying scan (GES) were utilized to access gastric function pre and post G-POEM. Post G-POEM follow up at 1, 3, 6, and 12 months was completed. PAGI-SYM was collected at all follow ups. For 6- and 12month visits, GES and EndoFLIP were conducted.
Results: Twenty-eight patients (age: 28-80, 82% female) underwent G-POEM between February 2018 to July 2019. All cases were done under general anesthesia with technical success rates of 100%. There were adverse events in 3/28 of the cases that were successfully managed endoscopically. There was significant improvement in gastric emptying post G-POEM (4-hour average retention rates of 6.3% compared to 35.0% pre-GPOEM, p < 0.001). There was a significant improvement in patient reported PAGI-SYM score from baseline (3.0) and 3months post G-POEM (0.80), p < 0.001. There was a trend towards sustained improvement in PAGI-SYM score on 6 month follow up, but this did not reach statistical significance (2.10, p = 0.22). There were improvements noted in pylorus cross sectional area and distensibility index on EndoFLIP post G-POEM, but this did not reach statistical significance and will require further assessment and a larger sample size.
Conclusion and Potential Impact: G-POEM appears to be a safe and feasible treatment alternative for gastroparesis with significant short-term improvements in gastric emptying rates and overall symptom scores. A larger cohort of patients is needed to validate these findings and examine the long-term effect of G-POEM on patients’ wellbeing and healthcare utilization.
Evaluating the Effect of Acarbose Treatment on Insulin Secretion and Sensitivity in Early Diabetes Using a Novel Interpretation of the Disposition Index Equation
Authors: Hanna Clarissa , Hannon Tamara , Considine Robert V. , Mather Kieren J.
Published: October 2019

Background and Hypothesis: In pathologic states such as obesity and insulin resistance, there is a progressive decline in insulin sensitivity requiring greater insulin secretion to maintain normoglycemia. The inverse relationship between insulin sensitivity and secretion is mathematically defined by the Disposition Index (DI), a measure of βcell function adjusted for insulin sensitivity. We are working to generalize the DI equation to allow direct physiologic interpretation of the DI term, and of the slope relating insulin secretion with insulin sensitivity. We tested study treatment effects hypotheses using these new analytic methods.
Background and Hypothesis: In pathologic states such as obesity and insulin resistance, there is a progressive decline in insulin sensitivity requiring greater insulin secretion to maintain normoglycemia. The inverse relationship between insulin sensitivity and secretion is mathematically defined by the Disposition Index (DI), a measure of βcell function adjusted for insulin sensitivity. We are working to generalize the DI equation to allow direct physiologic interpretation of the DI term, and of the slope relating insulin secretion with insulin sensitivity. We tested study treatment effects hypotheses using these new analytic methods.
Results: These analyses revealed statistically significant 1-year changes in DI, in secretion-sensitivity coupling slopes, and in the joint changes in secretion and sensitivity. However, these treatment effects did not differ by randomized treatment group, suggesting an on-study effect beyond the randomized treatments.
Conclusion and Potential Impact: 
We have applied a novel analytic approach to evaluate the secretion-sensitivity relationship modeled by the disposition index equation to investigate the effect of randomized therapy on β-cell function in a placebo-controlled randomized clinical trial. These analyses revealed study effects on the secretion-sensitivity relationship that have not been previously described, suggesting that this novel approach will have value in clinical studies of β-cell dysfunction and treatment effects.
Percutaneous Coronary Intervention Outcomes in Solid Organ Transplant Candidates
Authors: Harris, BS Kate L. , Frick, MD Kyle , Lee, MD Lawrence
Published: October 2019

Background: As part of the pre-transplant assessment, patients with end-stage renal, liver, pancreas, or lung disease who wish to attain transplant eligibility must undergo evaluation for coronary artery disease (CAD). Any significant CAD must be treated, usually by revascularization via either percutaneous coronary intervention (PCI) or coronary artery bypass grafting (CABG), in order to achieve transplant candidacy. PCI in these patients is inherently higher risk due to baseline comorbidities, but there are few studies reporting outcomes following PCI in this population. We sought to investigate the short- and intermediate-term outcomes in patients undergoing PCI as part of a transplant candidacy evaluation. We also aimed to assess whether these patients ultimately received the desired transplant after PCI.
Methods: This is a retrospective study investigating all patients who underwent PCI as part of a pre-transplant evaluation between 2009 and 2017 at IU Health Methodist Hospital. Patients were identified and data variables were extracted from an institutional American College of Cardiology CathPCI database. Medical records of all patients were reviewed to determine date of initial PCI and the type of solid organ transplant each patient was being evaluated for. Primary outcomes measured included 30-day and 1-year mortality, and whether organ transplantation ultimately occurred.
Results: A total of 497 patients were identified. Pre-transplant PCI performed in end-stage liver disease was most common (n=182), followed by renal (n=167), lung (n=74), multi-organ (n=66), pancreas (n=6), and intestinal (n=2). Combined 30-day mortality was 4.9%, 5.4%, 12.2%, 0%, 0%, and 0% for liver, renal, lung, multi-organ, pancreas, and intestinal, respectively. Combined 1-year mortality was 23.1%, 7.8%, 12.2%, 37.9%, 0%, and 0% for liver, renal, lung, multi-organ, pancreas, and intestinal, respectively. The percentage of patients ultimately receiving the desired transplant was low, with 32.4% for liver, 35.9% for renal, 32.4% for lung, 57.6% for multiorgan, 83.3% for pancreas, and 0% for intestinal.
Conclusion: This study demonstrates that PCI in patients undergoing solid organ transplant evaluation is relatively high-risk based on the 30-day and 1-year mortality. Furthermore, the percentage of patients ultimately receiving a transplant is relatively low. These results raise the question of whether high-risk PCI is the optimal CAD treatment in this population. These results also raise the question of whether changes to the transplant care protocol should be made to improve the likelihood of receiving a transplant before continuing to subject these patients to high-risk PCI.
Characterization of RNA binding proteins required for receptor-mediated endocytosis in C. elegans
Authors: Holdeman Margaret D. , Daouda Farida , Hundley, Ph.D. Heather A.
Published: October 2019

Background and Hypothesis: Recently, a study conducted by the Hundley lab revealed that mutation of a specific RNA editing protein, ADR2, in the model organism C. elegans increased receptor-mediated endocytosis of yolk protein vit-2 and oocyte maturation. Of interest to us was what additional RNA binding proteins were involved in the expression of vit-2. To this end, RNA interference (RNAi) was used to knock out relevant RNA binding proteins. We hypothesized we would find candidates that both enhanced and reduced the expression of vit-2.
Project Methods: Previously, adr-2(-) C. elegans strain expressing a yolk protein (vit-2) fused to GFP and a library of E. coli strains expressing RNAi directed to specific RNA binding proteins were engineered. vit-2:GFP reporter worms were grown on each RNAi bacterial strain. The COPAS Biosort, a large particle sorter which detects fluorescent intensity, was used to visualize and quantify vit-2 endocytosis.
Results: Our screen yielded ten candidate proteins. These proteins had various functions and were expressed in different tissues, most commonly the nervous and reproductive systems. Remarkably, proteins gld-1 and puf-8 are negative regulators of oocyte fate determination and spermatogenesis respectively. Unlike the rest of our candidates, mutation of these two proteins did not have an observed effect on developmental progression but still caused significant increase in vit-2 expression.
Conclusion and Potential Impact: This screen successfully gave direction to future studies of ADR-2 involvement in yolk protein uptake, oocyte maturation, and embryonic development. Two of our identified targets seem to be involved in reproductive processes. The remainder may be affecting vit-2 expression by altering development or receptor-mediated endocytosis. Our goal in the immediate future will be to compare loss of these target proteins in wildtype worms to loss in adr-2(-) worms. This would add to what is known about ADARs and life span and possibly demonstrate a role for ADARs in germ cell maturation.
Pediatric Anticholinergic Toxidrome and Treatment with Physostigmine
Authors: Huber Sarah , Avera, MD Bob , Overberg PharmD, BCPS, CSPI Adam , Morton, MPH Shannon , Nanagas MD Kristine
Published: October 2019

Background and Hypothesis: Physostigmine is the antidote to anticholinergic poisoning. Widely used in the 1970s, 2 cases published in 1980 associated use of physostigmine with significant adverse cardiotoxic events. This caused widespread opposition to the use of physostigmine. More recently, the safety profile has been re-examined, and it has been shown to have a more favorable safety profile than was previously believed. Current literature focuses on adult populations, and the available pediatric data does not comprehensively evaluate treatment plans for anticholinergic toxicity in pediatrics. We sought to establish the prevalence of pediatric anticholinergic toxicity as well as determine the prevalence of exposure types associated with physostigmine administration. We then looked at the difference in outcomes, including mortality, for those treated with physostigmine versus those treated with benzodiazepines.
Project Methods: We retrospectively analyzed data from the National Poison Data System (NPDS), a database collected from poison centers nationwide. We queried for all poison center exposure cases for ages 2-18 to selected anticholinergic agents or any cases that received physostigmine from January 1, 2013 – December 31, 2017.
Results: The NPDS had a total of 109,833 patients exposed to one of the selected anticholinergic agents or plants. Only 0.27% of cases were treated with physostigmine (n=298), versus 3.3% that were treated with benzodiazepines (n=3626). The most prevalent exposure was diphenhydramine. The most likely pediatric patients to be treated with physostigmine are those that are adolescents, exposed to diphenhydramine, or ingested the substance intentionally with suspected suicidal intent.
Conclusion and Potential Impact: Despite a good safety profile and superior efficacy to benzodiazepines, physostigmine is still under-utilized by physicians to treat patients with an anticholinergic toxidrome. Further study can also be carried out on the potential of physostigmine to reduce resource utilization in treating the anticholinergic toxidrome.
A Multi-Disciplinary Care Model for People with Drug-Resistant HIV: The AMPATH HIV Drug Resistance Clinic in Kenya
Authors: Jawed Bilal , Humphery John , Gardner Adrian
Published: October 2019

Background: HIV mortality has decreased with the availability of ART (antiretroviral therapy) in resource-limited settings, resulting in an increase in the number of people living with HIV (PLHIV) globally. This increase in PLHIV has shifted the HIV-care model from an acute to a chronic approach requiring more effort to combat barriers to viral suppression such as long-term adherence to ART, stigma, and drug resistance. An increasing number of PLHIV are failing “2nd line” protease-inhibitor based ART as a result of these barriers, leaving these patients with limited options for treatment. To address this issue, the Academic Model Providing Access to Healthcare (AMPATH) created a multidisciplinary HIV-drug resistance clinic focusing on supporting patients failing second-line ART in 2015. The objective of this study is to describe the implementation of the AMPATH DRC in western Kenya.
Methods: The HIV Drug Resistance Clinic (DRC) accepts patients from over 20 surrounding HIV clinics in a setting with over 100,000 PLHIV. The DRC identifies and enrolls patients failing second-line ART defined as ≥ 2 viral loads ≥ 1,000 copies/mL despite adherence interventions. The multidisciplinary team consists of HIV-specialist physicians, trained HIV-peer counselors, clinical pharmacists, and social workers who collaborate with patients in clinic to identify barriers to adherence and implement patient-centered interventions to mitigate barriers to adherence, treat drug resistance, and maximize the efficacy of ART. The DRC staff has expertise in ART regimen selection for patients with advance HIV drug resistance through analysis of drug resistant test results (i.e. HIV genotype). Over 600 patients have received care at the DRC since 2015.
Personal role: Identifying barriers and facilitators to implementation of DRCs in regional AMPATH sites, data abstraction to determine DRC clinical outcomes, and introduction of DRT results in the AMPATH MRS.
Moving Forward: By building the capacity of clinical leaders in decentralized regional AMPATH sites, we hope to expand the DRC care model within the AMPATH network.
Vision Dysfunction in Circadian Clock Gene Bmal Mice
Authors: Khan Humza , Mathew Deepa , Luo Qianyi , Bhatwadekar Ashay
Published: October 2019

Background and Hypothesis: The circadian rhythm disruption due to shift work results in a range of disorders such as metabolic disturbances, obesity, cardiovascular diseases, and insulin resistance. Interestingly, the core clock gene Brain and Muscle ARNT-Like 1 (Bmal1), is dysfunctional in shift workers. We reasoned dysfunctional Bmal will affect normal vision function. To do so, a genetically modified mouse with disrupted Bmal was assessed for visual function. We hypothesized that Bmal knockout mice will exhibit reduced retinal functions such as impaired acuity, accommodation, and tracking.
Experimental Design or Project Methods: The Bmal+/- mice were inbred and genotyped to obtain wild-types (WT), Bmal+/-, and Bmal-/-. To assess the retinal function, we performed electroretinogram (ERG) recordings at the zeitgeber times (ZT) of 0, 6, 12, and 18 which correspond to 7 AM, 1 PM, 7 PM, and 1 AM, respectively, under both scotopic and photopic conditions. The optokinetic response (OKR) assessments were measured in between ZT-3-ZT7.
Results: Consistent with previous studies, the ‘a’ wave and ‘b’ wave amplitudes of WT mice demonstrated a circadian rhythm under scotopic condition. There was a decrease in ERG amplitude of Bmal+/-, and Bmal-/- when compared to the WT group. Under photopic conditions, the circadian peak of ERG amplitude was reversed for Bmal-/- when compared to both WT and Bmal+/- mice. The OKR assessment was decreased substantially for Bmal+/- (0.3748c/d), and Bmal-/- (0.3130c/d) as compared to the WT mice (0.4827c/d). 
Conclusion and Potential Impact: Our studies demonstrate that the loss of Bmal leads to vision dysfunction possibly due to impaired rod and cone function. Furthermore, by using a mouse model of circadian rhythm dysfunction, we identified that individuals working on irregular shifts might be vulnerable to vision dysfunction, and our studies warrant timely testing of visual function and strategies for prevention of vision problems in shift workers.
Intraperitoneal Mesalamine Does Not Restores Mesenteric Perfusion or Prevent Mucosal Injury Following Intestinal Ischemia and Reperfusion
Authors: Knowles Thomas , Hosfield Brian , Shelley Chris , Markel Troy
Published: October 2019

Background and Hypothesis: Acute Mesenteric Ischemia (AMI) is characterized by a sudden decrease in blood flow to varying segments of the small intestine. It can lead to cellular damage, life-threatening intestinal necrosis and, if corrected, subsequent reperfusion injury. Reducing inflammation is key to preventing further cell damage. We therefore hypothesized that administration of mesalamine prior to intestinal ischemia would reduce epithelial cell damage by I/R and restore mesenteric perfusion.
Project Methods: C57Bl6J wild type (WT) mice were assigned to mesalamine or vehicle treatment groups (N=8/group). Prior to surgery, mice underwent intraperitoneal injection of treatment. Midline laparotomy was performed. Intestines were eviscerated, superior mesenteric artery (SMA) located, and baseline intestinal perfusion determined using Laser Doppler. SMA was then occluded to induce intestinal ischemia for sixty minutes, thereafter the occlusion was removed. Mesenteric reperfusion was then determined by Laser Doppler. Midline incisions were reapproximated with suture and animals were allowed to recover. After twentyfour hours, animals were re-anesthetized and underwent final assessment of mesenteric perfusion by Laser-Doppler. Animals were then euthanized, and intestines explanted. A portion of tissue was snap frozen for assessment for proinflammatory cytokines by ELISA. Another portion of tissue was stained with H&E and scored for intestinal mucosal injury. Data were assessed for normalcy and compared by Mann-Whitney-U test. P<.05 was significant.
Results: Preliminary data suggests mesalamine treated mice show no significant change in mortality compared to vehicle. Mesalamine treated mice also show an insignificant increase in histological damage score. Despite this, they show an insignificant improvement in oxygen perfusion.
Conclusion: Intraperitoneal mesalamine administration does not appear to be a useful method for limiting cell damage in GI diseases associated with AMI such as necrotizing enterocolitis. A larger sample size is needed to further elucidate treatment effects.
Role of perilipins in the development of fatty liver disease.
Authors: Kubascik Erica , DeGoey Madison , Gupta Dr. , Kostrominova Dr.
Published: October 2019

Background and Hypothesis: Lipid droplets (LDs) are fatty acid (FA) containing structures within cells. In obesity, hepatic LDs accumulate more FA which may cause steatosis and nonalcoholic fatty liver disease (NAFLD). Perilipins (PLINs) are a family of LDassociated proteins involved in intracellular trafficking and signaling. In hepatocytes, NAFLD alters expression of PLINs. Using metabolomics, Dr. Gupta’s laboratory previously showed enlarged LDs and altered PLIN1, 3, and 4 content in NOD2-/- compared to wild type (WT) mice on high fat diet (HFD). Nucleotide-binding and oligomerization domain (NOD) is an intracellular receptor that regulates sensitivity to obesity.
To expand on Dr. Gupta’s study, we quantified diet-induced alterations and visualized intracellular distribution of PLINs in mouse livers. We hypothesized that PLIN2, 3, and 4 concentrations would increase whereas PLIN1 would decrease in NOD2-/-HFD compared to WTHFD mice.
Methods: Immunostaining was used to visualize intracellular distribution of PLINs. Western blotting was used to quantify differences in PLINs protein expression.
 Results: LD distribution showed WT regular chow (RC) = NOD2-/-RC < WTHFD<<NOD2-/-HFD. Bright LD-associated staining for PLIN2 was observed in both small and large LDs in all four groups. PLIN3 brightly stained the bile ducts, and it stained small LDs but not large LDs. PLIN4 stained small intracellular LDs in all four groups. PLIN1 showed a trend for decrease in both NOD2-/-HFD and WTHFD compared with RC mice. PLIN2 appears to be decreased in WTHFD and both NOD2-/- groups compared to WTRC. PLIN3 seemed to show increased expression in WTHFD but not in NOD2-/-. PLIN4 showed a trend for decreased expression in both NOD2-/-RC and NOD2-/-HFD mice compared to WTRC.
Conclusion: Despite enlarged LD size, there was no detectable increase in PLINs expression in NOD2-/-HFD compared to WTHFD. This may be due to the impact of other LDassociated proteins in the livers of NOD2-/- mice.
The Effects of Propofol on the Human Blood Brain Barrier
Authors: Lewis Kirsten A. , Hughes Jason M. , Canfield Scott G.
Published: October 2019

Background and Hypothesis: Recently, the safety of repeated and lengthy anesthesia in young children has been called into question. Previous studies have shown propofol, an anesthetic, can diminish blood-brain barrier (BBB) properties. However, the underlying cellular mechanisms are relatively unknown. The BBB is critical in ensuring that potentially harmful circulating factors are impermeable to the brain. Previous animal studies models have shown that propofol increases the levels of matrix metalloproteinases (MMPs), which have independently been shown to degrade the extracellular matrix and breakdown tight junctions, a critical component of the BBB. Hypothesis: Propofol exposure to a human induced pluripotent stem cell (iPSC)-derived BBB model increases MMP activity ultimately contributing to a leaky barrier phenotype.
Experimental Design or Project Methods: This study utilized human iPSCs differentiated into brain microvascular endothelial cells (BMECs), the barrier forming cell type of the BBB. iPSC-derived BMECS were exposed to propofol (50μM) for three hours and barrier properties were monitored. Barrier tightness was monitored using trans-endothelial electrical resistance (TEER) and sodium fluorescein permeability assays. Tight junction localization was determined with immunocytochemistry. MMP activity was determined with SensoLyte assay kits. To determine the role of MMPs, a broad spectrum MMP inhibitor, GM6001, was utilized and barrier properties were monitored.
Results: Propofol treatment significantly reduced TEER and increased sodium fluorescein permeability, indicative of a leaky barrier. Propofol treatment increased levels of MMP-2 activity but not MMP-9 when compared to non-treated BMECs. Inhibition of MMPs by GM6001 prior to propofol treatment appeared to partially restore barrier integrity as monitored by sodium fluorescein permeability.
Conclusion and Potential Impact: These results indicate that increased MMP-2 activity levels could be in part responsible for diminished BBB properties. Inhibition of MMPs protected barrier integrity from propofol treatment. A further understanding of the underlying mechanisms of anesthetic-induced damage can potentially improve anesthesia safety.
Patient-Reported Outcomes with Tibial Baseplate Position and PCL Status in Conforming Total Knee Arthroplast
Authors: Madden, BS Joseph A. , Arnold, MS Payton K. , Deckard, BSE Evan R. , Meneghini, MD R. Michael
Published: October 2019

Background and Hypothesis: Anteroposterior (AP) tibial baseplate position, posterior tibial slope, and posterior cruciate ligament (PCL) status in total knee arthroplasty (TKA) undoubtedly affect kinematic patterns. Further, few studies document patient-reported outcome measures (PROMS) in conforming polyethylene TKA with these varying parameters. The purpose of this study was to correlate PROMS with the interaction of AP tibial baseplate position, tibial slope, and PCL status in a consecutive series of primary TKAs with conforming polyethylene. We hypothesized that pain, function, and satisfaction may correlate with a combination of these three parameters.
Experimental Design or Project Methods: 589 consecutive primary TKAs of a single implant design performed by a single surgeon between January 2016 and October 2018 were retrospectively reviewed. AP tibial baseplate position (relative to the middle of the tibial canal) and posterior tibial slope measurements were performed on 4-week postoperative sagittal view radiographs with a standardized measurement protocol by two independent blinded raters. Validated PROMS related to activity level, pain, and function were evaluated at minimum one-year.
Results: Analyses indicated differences based on ≥8mm vs. <8mm of posterior distance of the tibial baseplate from the tibial canal and whether or not the PCL was released. Four analysis groups (PCL-preserved <8mm, PCL-preserved ≥8mm, PCL-resected <8mm, and PCL-resected ≥8mm) did not differ by demographics/covariates (p≥0.150), tibial slope (p≥0.132), or preoperative PROMS (p≥0.088). The PCL-released <8mm group achieved clinically meaningful higher absolute (92.0) and delta (42.0) median KOOS Jr. scores, higher satisfaction (96.3%), and the greatest reduction in pain while climbing stairs (-7.0) although some findings lacked statistical significance.
Conclusion and Potential Impact: In conforming polyethylene TKAs, releasing the PCL in combination with AP tibial baseplate placement <8mm from the tibial canal may eliminate kinematic conflict between the PCL and tibial baseplate leading to improved satisfaction, function, and pain while climbing stairs.
Thrombopoietin: Role in Fracture Healing and Pain
Authors: McVeigh Luke G. , Babu Deepa Shiek Pran , Blosser Rachel J. , Thompson Eric L. , Fehrenbacher Jill C. , White Fletcher A. , Kacena Melissa A.
Published: October 2019

Background and Hypothesis: The megakaryocyte (MK) growth factor, thrombopoietin (TPO), improves bone healing in mice, rats, and pigs. Here we explore the role of MK-secreted platelet-derived growth factor-BB (PDGF–BB) in TPO’s mechanism of improving fracture healing. Interestingly, PDGF has neuroprotective effects, and neuronal PDGF receptors (PDGFRs) are primarily stimulated by PDGF–BB. Additionally, fibroblast growth factor 2 (FGF2) is secreted by osteoblasts (OBs), and MKs stimulate OB proliferation; therefore, MKs may regulate FGF2 expression which also promotes nerve regeneration. Importantly, neuropathic pain caused by fractures can be prevented by neuroprotective therapies. We hypothesize that increases in fracture-related neuropathic pain observed with age are due to reductions in: MK-secreted PDGF-BB and MK-stimulated, OBsecreted FGF2.
Experimental Design or Project Methods: Dorsal root ganglia (DRG) and MKs from bone marrow and spleens were isolated from 3-4 month-old (young) and 22-24 month-old (old) mice. MKs were also isolated from E13-15 fetal livers of mice and OBs were isolated from the calvaria of neonatal mice. OBs were cultured alone or in the presence of MKs for 4 days. Real-time PCR was completed to examine the expression of PDGF-BB in MKs, FGF2 in OBs, and PDGFR in DRG.
Results: Old MKs exhibit a 63% reduction in PDGF–BB expression and old DRG exhibit a 68% reduction in PDGFR expression. OBs cultured with MKs show a 1.78 fold increase in FGF2 expression.
Conclusion and Potential Impact: Decreased expression of PDGF-BB in old MKs and PDGFR in old neurons, as well as increased FGF2 expression in OBs cultured with MKs present possible mechanisms for both the reduction in bone healing and increased fracture pain observed with age. TPO improves bone healing and may potentially reduce neuropathic pain directly by increasing MK secreted PDGF-BB and indirectly through MK PDGF-BB stimulating FGF2 secretion from OBs.
Health Care System Distrust, Race, and Surrogate Decision Making Regarding Code Status
Authors: Na, BS, MS Sang Yoon , Slaven, MS James E. , Burke, BA Emily S. , Torke, MD, MS Alexia M.
Published: October 2019

Background and Hypothesis: Studies have shown African American patients are more likely to prefer aggressive life-sustaining treatments such as cardiopulmonary resuscitation (CPR) at end-of-life compared to non-Hispanic White patients. Given prior racial disparities in healthcare, low trust has been proposed to explain these preferences. We examined factors that influence surrogate decision makers’ preference for Do Not Resuscitate (DNR) status for hospitalized older adults who cannot make their own medical decisions. We explored whether race is associated with surrogate preference for DNR status for a hospitalized older adult. We also examine if race is associated with distrust and if the race/code status relationship is partially explained (mediated) by distrust in the healthcare system.
Experimental Design or Project Methods: Analyses were conducted using data from an observational study of patient/surrogate dyads admitted to an ICU in a Midwest metropolitan area. Distrust was assessed using the Revised Health Care System Distrust Scale. A single item asked the surrogate which status they thought was best for the patient, full code or DNR.
Results: In bivariate analysis, higher proportion of African American surrogates showed preference for full code (62.4% vs 37.6%, p=0.0001). After adjusting for trust and sociodemographic and psychological covariates, race was still significantly associated with DNR preference (aOR = 1.92; 95% CI: 1.04, 3.55; p=0.0382). Surrogate race did not show significant association with distrust in bivariate or multivariable analysis, which adjusted for sociodemographic and psychological covariates (p=0.3867).
Conclusion and Potential Impact: Contrary to previous studies, we observed no association between surrogate race and distrust of the health care system. Differences in code status preference may be due to other factors related to race and culture. In order to ensure patients are receiving end-of-life care that is consistent with their values, more work is needed to understand the cultural complexities behind end-of-life care preference.
GENERATION OF MOSAIC STEM CELL DERIVED INNER EAR ORGANOIDS TO DETERMINE PARACRINE EFFECTS OF TMPRSS3
Authors: Nabrzyski, BS Radoslaw , Tang, PhD Pei-Ciao , Alex Alpha , Nelson, MD, PhD Rick F.
Published: October 2019

Background and Hypothesis: The carefully timed treatment of mouse embryonic stem cell (mESC) cultures with small molecules results in mESC differentiation into 3D organoids containing all components of the inner ear such as hair cells (HCs), support cells and neurons. Loss of TMPRSS3 function, which is a transmembrane extracellular protease, has been previously shown to lead to rapid HC degeneration between culture days 36 (D36) and D38 in 3D organoids. Mosaic organoids would allow for analysis of developmental dynamics, cell-cell interactions and even therapeutic rescue efficiency. We hypothesized that we could develop an inner ear mosaic organoid containing cells with and without TMPRSS3 and that the Tmprsss3KO mosaic organoids would have greater hair cell survival than Tmprss3KO-only organoids due to the compensatory effect of intact TMPRSS3 on control hair cells within the same vesicle.
Experimental Design or Project Methods: Two mESC cell lines (R1E background) were previously generated using CRISPR-Cas9n. Control reporter line (tdTomato) expresses a tdTomato reporter gene in all cells under the CAG promoter at the ROSA26 locus. Tmprss3 knockout line (Tmprss3KO) contains a 2A-nGFP cassette with a premature stop codon into exon 2 of the Tmprss3 gene. We generated Tmprss3KO mosaic (Tmprss3KO:tdTomato) and control mosaic (R1E:tdTomato) organoids. The aggregates were analyzed on days 25, 33 and 38 after being fixed, sectioned and stained for tdTomato, SOX2, MYO7A, cleaved CASPASE3, and BK channels using immunohistochemistry.
Results: We have successfully used mESCs to generate Tmprss3KO mosaic inner ear organoids with similar efficiency to that of control mosaic organoids. Preliminary data suggests that the hair cell survival rates were similar across all vesicle types in the Tmprss3KO mosaic organoids. Additionally, the Tmprss3KO mosaic organoids had a significantly decreased overall BK channel expression by D38.
Conclusion and Potential Impact: The successful generation of mosaic organoids achieved here under both conditions sets the stage for future studies of intercellular interactions and therapeutics in this domain. More replicates are necessary to make a definitive conclusion about the effectiveness of control cells on Tmprss3KO rescue.