Proceedings of IMPRS http://journals.iupui.edu/index.php/IMPRS <p>The&nbsp;Indiana Medical Student Program for Research and Scholarship (IMPRS) facilitates IU School of Medicine medical student participation in various medical research and experiential opportunities including laboratory, clinical, and health research outcomes and community health education. The program strives to make a wide variety of research and clinical opportunities accessible to all students, enhance intellectual inquisitiveness, and support excellence in the development of physicians, physician-scientists, and educators.</p> en-US imprs@iupui.edu (Anne Nguyen) dapolley@iupui.edu (Ted Polley) Wed, 09 Oct 2019 00:00:00 -0400 OJS 3.1.2.4 http://blogs.law.harvard.edu/tech/rss 60 Multi-Modal MR/PET Imaging of Natural Killer Cell Immunotherapy Against Glioblastoma with Correlated Histology http://journals.iupui.edu/index.php/IMPRS/article/view/23404 <p><strong>Hypothesis:</strong> Immunotherapies hold great promise for treatment of highly resistant cancers, such as glioblastoma (GBM). We hypothesized that high powered imaging can be effectively combined to quantitatively assess the therapeutic efficacy of human derived natural killer (hNK) cells in an orthoptic xenografted mouse model of GBM.</p> <p><strong>Methods:</strong> Tumor take (TT) was established via fluorescence. Mice in the treatment (n=5) and control (n=4) groups were given IV hNK cells and physiological saline, respectively. MRI and PET scans were performed four and six weeks after tumor implantation. Histological slices were taken at time of death. Software analysis of tumor volume, standardized uptake value (SUV), and tumor-to-brain ratio (TBR) was conducted via Qimage and Indica Labs - HALO.</p> <p><strong>Results:</strong> Mean growth rates are as follows: T1 volume (mL) – 4.1 in the control group vs 2.3 in treatment. T2 volume (mL) – 6.0 in control vs 2.7 in treatment. PET volume (mL) – 3.1 in control vs 2.1 in treatment, SUV (g/mL) – 5.4 in control vs 3.0 in treatment. Two-tailed t-test analysis showed statistical significance (p &lt; 0.01) in T1 volume data.</p> <p><strong>Conclusion and Potential Impact:</strong> Treatment group mice showed a trend in reduction in growth rate of tumor volume and SUV compared to control, with a correlated lower histology TBR, suggesting effective in vivo assessment of hNK therapeutic efficacy in the mouse model of GBM via MR/PET imaging. Future trials should provide a larger population size to increase reliability, precision and power.</p> JM Acchiardo, YH Yun, S Smiley, P Marcadis, GD Hutchins, MC Veronesi Copyright (c) 2019 JM Acchiardo, YH Yun, S Smiley, P Marcadis, GD Hutchins, MC Veronesi https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23404 Tue, 08 Oct 2019 10:18:57 -0400 Papillary Pathology in Calcium Oxalate Stone Formers Who Do and Do Not Form Their Stones on Randall’s Plaque http://journals.iupui.edu/index.php/IMPRS/article/view/23407 <p><strong>Background and Hypothesis:</strong> Nephrolithiasis currently affects about one-in-eleven people with a recurrence in up to half of those individuals. Formation of calcium oxalate (CaOx) stone is most common. Some CaOx stones are known to form on Randall’s plaque, which is a calcification of the renal papilla, and this kind of stone can be identified by morphology. We divided CaOx stone formers into two groups: Randall’s plaque (RP) stone formers (RPSF) and non-Randall’s plaque stone formers (NRPSF). We hypothesized that renal papillary pathologies would be different between these two groups of stone formers.</p> <p><strong>Experimental Design or Project Methods:</strong> Surgical videos were assessed for papillary pathology using a semiquantitative grading system to measure papillary appearance in terms of ductal plugging and dilation, tissue surface pitting, loss of papillary contour, and RP. The second measure computed the papillary percent surface area of Randall’s plaque and ductal plugging using still images of the papilla. The scoring and quantitative measures of the papillae were compared between the two patient groups. All work was done in a manner blinded to the patient group.</p> <p><strong>Results:</strong> Two-tailed t-test showed that RPSF group had higher scores of pitting and RP and lower scores for plugging when compared to the NRPSF group. Similarly, the quantitative data showed that RPSF group had a lower percentage of plugging surface area and higher percentage of RP surface area.</p> <p><strong>Conclusion and Potential Impact:</strong> These data show that persons forming their CaOx stones primarily on RP have a papillary pathology that differs from CaOx stone formers who make their stones by other mechanisms. RPSF have more RP and less ductal plugging. Since the underlying pathologies existing in RP stone formation are different from other CaOx stone formers, it is possible that certain treatments could be especially effective for this group, and thus these results suggest that clinical trials that separate out this group of CaOx stone formers are warranted.</p> Haider N. Al-Awadi, BS, James C. Williams, Jr., PhD Copyright (c) 2019 Haider N. Al-Awadi, BS, James C. Williams, Jr., PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23407 Thu, 10 Oct 2019 09:55:11 -0400 Identification of Microbes Associated with the Urethra during Health and Inflammation http://journals.iupui.edu/index.php/IMPRS/article/view/23408 <p>The number of cases of urethritis, or inflammation of the male urethra, in the US has been estimated to be 2.8 million each year in the United States.1 It is the most common reason young men seek primary care and this syndrome is associated with acute proctitis, epididymitis, and orchitis.2 Approximately half of the cases are idiopathic, meaning that a causative agent cannot be identified. The goal of my project is to identify and characterize novel microorganisms that may be associated with male idiopathic urethritis. We used a variety of approaches to cultivate microorganisms from archived urethral swabs collected from men with idiopathic urethritis and healthy controls. We defined the phylogeny of the isolates using 16S rRNA sequencing to identify organisms that are not presently represented in microbial databases. Currently, we are scaling up a number of novel taxa we identified for genome sequencing. By adding these new genome sequencing to the existing databases we will be able to assign a higher proportion of reads from corresponding metagenomic sequence data and achieve a more complete survey of the male urethral microbiota in health and urethritis. This will provide crucial information which may permit us to identify urethritis associated organisms, develop new molecular diagnostics for these organisms, and discern if how these organisms are sexually transmitted </p> <p>References <br> 1. Brill JR. Diagnosis and treatment of urethritis in men. Am Fam Physician 2010; 81:873–8. 2. Bachmann LH, Manhart LE, Martin DH, Sena AC, Dimitrakoff J, Jensen JS, et al. Advances in the Understanding and Treatment of Male Urethritis. Clin Infect Dis. 2015;61 Suppl 8:S763–S9.</p> Jonathan Alessi, Rowan Farrell, Evelyn Toh, Xiaoli Zhang, David E. Nelson, Stephen J. Jordan Copyright (c) 2019 Jonathan Alessi, Rowan Farrell, Evelyn Toh, Xiaoli Zhang, David E. Nelson, Stephen J. Jordan https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23408 Tue, 08 Oct 2019 10:20:12 -0400 Community perspectives on early childhood development in Kenya: A qualitative study. http://journals.iupui.edu/index.php/IMPRS/article/view/23409 <p><strong>Background and Hypothesis:</strong> Kenyan children are at risk for having delayed early childhood development (ECD). The Care for Child Development (CCD) is a program created by UNICEF for the promotion of ECD by increasing caregiver’s knowledge about attitudes, nutrition, communication, playing, and physical health. The objective of this study was to assess key informants’ and caregivers’ perspectives on ECD and the introduction of this CCD-based intervention within the community, as well as caregivers’ perspectives on ECD after receiving the intervention. We hypothesize that caregivers who implement CCD practices into their daily life will have increased nurturing interactions with their children and positively influence their child’s development.</p> <p><strong>Experimental Design or Project Methods:</strong> This study used semi-structured interviews of key informants and focus group discussions of caregivers to elicit perspectives on ECD and a CCD-based intervention in Eldoret, Kenya. The intervention was a curriculum adapted from the CCD program administered within 10 mother-group sessions. Caregivers were interviewed both before and after the intervention, while key informants were interviewed at one time-point. All interviews and focus groups were audio-recorded, transcribed and translated. Qualitative analysis was performed using Dedoose software. </p> <p><strong>Results:</strong> Collectively, community members and caregivers believed the relationships a child has with their caregiver, family, and community greatly influences their development. In addition, they believed emotional burden and social stigma are challenges caregivers experience and this limits their ability to take their child to necessary medical services. The intervention group appreciated the parental ECD education and further disseminated the information to other caregivers and members in their community. </p> <p><strong>Conclusion and Potential Impact:</strong> By understanding the community’s collective views, we can implement the CCD program more effectively in Kenya and empower caregivers with the knowledge on how to create an environment that stimulates ECD.</p> Brianna Alex, Catherine Raciti, Dr. Megan McHenry Copyright (c) 2019 Brianna Alex, Catherine Raciti, Dr. Megan McHenry https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23409 Tue, 08 Oct 2019 10:20:57 -0400 Is sedentary behavior associated with dysglycemia in youth with obesity? http://journals.iupui.edu/index.php/IMPRS/article/view/23411 <p><strong>Background and Hypothesis:</strong> The childhood obesity epidemic is linked with an increase in dysglycemia and type 2 diabetes (T2D) amongst youth.<sup>i</sup> Adolescence is associated with decreased levels of physical activity<sup>ii</sup>, however, there is a paucity of research investigating physical activity measures and dysglycemia in youth. We hypothesize that decreased physical activity and/or increased sitting time is positively correlated with dysglycemia.</p> <p><strong>Project Methods:</strong> Study participants were youth aged 10-21y with a BMI &gt;85th percentile for age and gender. Accelerometers (activPAL) and the FELS Physical Activity Questionnaire (FELS PAQ) were used to assess objective and selfreported physical activity levels. Glucose tolerance was assessed with 2-hour oral glucose tolerance tests (OGTT). Independent t-tests were used to compare physical activity levels for participants with normal glucose tolerance (NGT) and dysglycemia. Correlation analysis was performed to evaluate relationships between measures of physical activity and OGTT measures. </p> <p><strong>Results:</strong> Participants with NGT (N=27) and dysglycemia (N=26) had comparable demographics; age, race, and ethnicity. Hemoglobin A1c (HbA1c) (p =0.002), average fasting glucose (p&lt;0.00), indices of insulin resistance (HOMA-IR) (p=.023), insulin secretion (disposition index, DI, a marker of risk for T2D) (p=.008), 2-hour-OGTT measures for glucose (p=.004) and insulin (p=.002) differed between groups. There were not group differences for objective or selfreported measures of physical activity. For the entire cohort, sitting time was positively associated with OGTT 2hr glucose (r=.38, p=.03). However, a subgroup analysis showed that the association between sitting time and OGTT 2hr glucose was significant in the dysglycemia group only (r =.64, p=.006, vs r =.016, p=.95 for NGT group). Self-reported measures of activity (Likert scores) and OGTT measures for insulin and glucose were positively correlated.&nbsp;</p> <p><strong>Conclusion and Potential Impact:</strong> Increased sitting time is associated with impaired glucose tolerance in youth with obesity at risk for T2D.</p> <p><br><sup>i</sup>Van der Aa MP, Fazeli Farsani S, Knibbe CA, de Boer A, van der Vorst MM. Population-Based Studies on the Epidemiology of Insulin Resistance in Children. J Diabetes Res. 2015;2015:362375.</p> <p><sup>ii</sup>Dumith SC, Gigante DP, Domingues MR, Kohl HW. Physical activity change during adolescence: a systematic review and a pooled analysis. International Journal of Epidemiology. 2011;40(3):685-98. </p> <p>&nbsp;</p> Alan Alvarez de Sotomayor, BS, Hala El-Mikati, MD, Lisa Yazel-Smith, MS, MCHES, Tamara Hannon, MD, MS Copyright (c) 2019 Alan Alvarez de Sotomayor, BS, Hala El-Mikati, MD, Lisa Yazel-Smith, MS, MCHES, Tamara Hannon, MD, MS https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23411 Tue, 08 Oct 2019 10:21:26 -0400 The effects of NK cells and drug combination therapy on glioblastoma tumor growth http://journals.iupui.edu/index.php/IMPRS/article/view/23415 <p><strong>Background and Hypothesis:</strong> Glioblastoma (GB) is an aggressive primary brain malignancy with a mean survival time of 15-16 months. Initial therapy is limited to surgery, radiation and temozolamide (TMZ) chemotherapy with up to seventy percent of GBs recurring in the first year and only five percent of patients still alive after five years. A reason these tumors are difficult to treat is the effect that the GB tumor microenvironment has on recurrence. One major cell type in this microenvironment is GB cancer stem cells and these cells play a major role in recurrence of the GB tumor. Our approach is that if we reduced tumor volume with the standard of care and treat with NK-cell based treatment and combination drug therapy we can improve the remission and prognosis of GB tumors.</p> <p><strong>Experimental Design or Project Methods:</strong> <br> To explore the killing potential of NK cells as well as combination drug treatment we set up a caspase assay that would indicate DNA damage and cell health. We set up in a 96-well plate with GB43 or Cancer stem cells with increasing consideration of either drug over 72 hours or NK cells over 48 hours. After we create, dose curves to find LD50 of both Cell based and drug based treatment. </p> <p><strong>Results:</strong> <br> We are still assessing the cell-based therapy and calculating LD50 for both treatments. However, we have found that in the drug based combination therapy we were able to show that paclitaxel and RG3788 both alone and in combination with the standard of care,TMZ, showed significantly reduced GB43 cell viability as concentration increased at the 72 hour time point. However, cancer stem cell remain at around 80% viability across all drug combinations and concentrations.</p> <p><strong>Conclusion and Potential Impact:</strong> In conclusion at this point these data indicate that GB43 with the standard of care TMZ in combination with paclitaxel and RG3788 have the ability to kill cells in vitro. Moving forward we plan to package paclitaxel and RG3788 in nanoparticles for more efficient targeting of cells to target tumors. In addition, we plan to create lethal dose curves for NK cells and control neuronal cell tissue to better assess drug toxicity and efficacy.</p> Matthew Anderson, Jianyun Liu, Yeonhee Yun, Shelby Smiley, Michael Veronesi Copyright (c) 2019 Matthew Anderson, Jianyun Liu, Yeonhee Yun, Shelby Smiley, Michael Veronesi https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23415 Tue, 08 Oct 2019 10:23:37 -0400 The efficacy of P.O.P. on Teen violence and the anti-violence programing in Northwest Indiana and greater Chicagoland area http://journals.iupui.edu/index.php/IMPRS/article/view/23585 <p><strong>Background:</strong> 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. <br> 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. <br>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?</p> <p>Hypothesis:</p> <ul> <li>&nbsp;There is an increase in the interest in college/STEM professions.</li> <li>&nbsp;There is a decrease in incidences of violence at schools with active SAVE chapters or participating in an anti-violence event</li> </ul> <p><strong>Experimental Design or Project Methods:</strong> Surveys, Observations, Student Interviews, Intervention, Cohort Study, Program evaluations, Longitudinal Study<em> (Go to each school nearby and request the info)</em> <br> <br><em>The study’s population includes students from 7th grade through high school seniors located throughout 13 schools in the northwest Indiana region. POP</em></p> <p><strong>Results:</strong> <em>Analyze Survey Data and interviews for positive correlation with the data.</em>&nbsp;</p> <p><strong>Conclusion/Potential Impact:</strong> 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.</p> Isaiah Sloss, Chiamara Anokwute, Reuban Rutland, MD, Michael McGee, MD Copyright (c) 2019 Isaiah Sloss, Chiamara Anokwute, Reuban Rutland, MD, Michael McGee, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23585 Tue, 08 Oct 2019 00:00:00 -0400 Predictors of Successful Outcomes Following Revision Total Knee Arthroplasty for Flexion Instability http://journals.iupui.edu/index.php/IMPRS/article/view/23416 <p><strong>Background and Hypothesis:</strong> Flexion instability is a poorly understood cause of total knee arthroplasty (TKA) failure, often referred to as a “wastebasket” diagnosis for unexplained persistent pain. It remains a challenging diagnosis, with unpredictable outcomes following revision surgery. This study identified predictors of successful patient reported outcome measures (PROMS) following revision for flexion instability.</p> <p><strong>Methods:</strong> 121 consecutive revision TKAs for flexion instability from 2011-2018 at a single center were retrospectively reviewed. PROMS were prospectively obtained preoperatively and at minimum one-year follow-up. 33 potential predictors of PROMS encompassed: presenting symptoms, demographics/medical covariates, intraoperative observations, and pre/postoperative radiographic measurements. Variables related to outcomes with p≤0.20 in univariate analysis were entered in regression models. Nonsignificant predictors were pruned until only predictors with p&lt;0.05 and the best model statistics were achieved. </p> <p><strong>Results:</strong> The sample was 63% female with mean age of 65±11 and BMI of 34±7 kg/m2. Pre- to postoperative changes in three radiographic measurements vital to the stability and function of TKA were related to function (p&lt;0.001), pain (p=0.019), and satisfaction (p=0.030) after surgery. Pain with walking (p≤0.059) and stair climbing (p≤0.048) were almost exclusively related to demographic/medical covariates. Only one of the presenting symptoms commonly associated with flexion instability influenced early revision outcomes (p=0.021).</p> <p><strong>Conclusion and Potential Impact:</strong> Instability is one of the most common reasons for TKA failure leading to costly revision surgery. To our knowledge, this is the largest flexion instability cohort studied to date, including substantially more predictors in an attempt to elucidate factors indicative of successful treatment outcomes. With the exception of radiographic parameters reflecting surgical decisions, findings suggest that many factors related to outcomes may be beyond the control of arthroplasty surgeons.</p> Payton K. Arnold, Joseph A. Madden, Mary Ziemba-Davis, R. Michael Meneghini, MD Copyright (c) 2019 Payton K. Arnold, Joseph A. Madden, Mary Ziemba-Davis, R. Michael Meneghini, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23416 Tue, 08 Oct 2019 10:24:44 -0400 Development of 3D human kidney model using Urine-Derived Epithelial Cells and HUVECs http://journals.iupui.edu/index.php/IMPRS/article/view/23418 <p><strong>Background and Hypothesis:</strong> Development of realistic, scaffold-free 3D organoids/tubuloids such as renal-like tissue, can serve as models for study of renal drug toxicity/screening, tissue repair, and renal disease. Previous studies utilized urine-derived epithelial cells (UDEpCs) to create viable tubuloid structures suspended in 3D Matrigel, displaying renal epithelial gene expression and functionality. Alternatively, renal proximal tubule epithelial cells (PTECs) with HUVECs were co-cultured in a monolayer to improve the model. Therefore, we hypothesized that the addition of HUVECs will help to establish and optimize growth conditions of UDEpCs in a 3D environment.</p> <p><strong>Experimental design:</strong> UDEpCs were isolated and co-cultured with tdTomatoHUVECs in Matrigel with different combinations of growth media. Cell growth and endothelial cell tube formation were examined daily. PTECs and tdTomatoHUVECs were also used to make spheroids in low-binding plates. Real-time PCR analysis was performed to examine gene expression for epithelial markers (ABCC4, PAX8), an endothelial marker (PECAM1), and TGF-β in spheroids.</p> <p><strong>Results:</strong> A single cell-derived colony was isolated from human urine and was double positive for CD10 and CD13, two renal PTEC markers. Co-culture of UDEpCs and tdTomato-HUVECs in Matrigel dramatically increased the growth, survival, and tubuloid formation of both cell types, for up to 12 days. Total RNA from spheroids showed a decline over time in all conditions. Gene expression was decreased in co-culture conditions, but patterns of gene expression were maintained.</p> <p><strong>Conclusions and Potential Impact:</strong> Future studies will examine gene expression in Matrigel cells, further optimize conditions for growth of UDEpCs, and perform functional assays, such as brush border enzyme activity. Improved methods to develop 3D kidney organoids/tubuloids will produce more accurate models for renal function studies. Developing a functional model mimicking in vivo processes can lead to new applications for creating novel therapies and treatment strategies.</p> Aatif Basher, MS, William Goggins, BS, Erika Gramelspacher, BS, Julia Walsh, Mark H. Kaplan, PhD, Wenjun Zhang, PhD, Bruce Molitoris, MD, Mervin Yoder, MD, Burcin Ekser, MD, PhD, Ping Li, PhD Copyright (c) 2019 Aatif Basher, MS, William Goggins, BS, Erika Gramelspacher, BS, Julia Walsh, Mark H. Kaplan, PhD, Wenjun Zhang, PhD, Bruce Molitoris, MD, Mervin Yoder, MD, Burcin Ekser, MD, PhD, Ping Li, PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23418 Tue, 08 Oct 2019 10:25:41 -0400 PedsQL for Prediction of Postoperative Patient-Reported Outcomes following Chiari Decompression Surgery http://journals.iupui.edu/index.php/IMPRS/article/view/23419 <p><strong>Background and Hypothesis:</strong> Chiari malformation type I (CM-I) is a common pediatric disorder in which the cerebellar tonsils herniate&nbsp;the foramen magnum with associated spinal symptoms. The purpose of this study is to determine if the preoperative Pediatric Quality of Life Inventory (PedsQL) score is predictive of intermediate-term PedsQL outcomes following Chiari decompression surgery. The utility of preoperative patient-reported outcomes (PROs) to postoperative PROs has been previously demonstrated in adult spine surgery. However, to our knowledge, there is currently no widely accepted tool to predict short-, intermediate-, or long-term outcomes after pediatric Chiari decompression surgery.</p> <p><strong>Project Methods:</strong> A prospectively maintained database was retrospectively reviewed. Patients who underwent first-time decompression for symptomatic Chiari malformation were identified and grouped by&nbsp;their preoperative PedsQL scores: mild disability (score 80–100), moderate disability (score 60–80); and severe disability (score &lt; 60). PedsQL outcomes were collected at 6-week, 3-month, and 6-month follow-up. Preoperative PedsQL subgroups were tested for an association with demographic, perioperative characteristics, and improvements between groups by one-way ANOVA, chi-square, or Wilcoxon signed rank test, where appropriate.</p> <p><strong>Results:</strong> The postoperative patient- and parent-reported PedsQL values were significantly different between all three groups at 6-weeks, 3-months, and 6-months after surgery (p &lt; 0.05), except at the 6-month timepoint for parent-reported outcomes (p = 0.111). Patients with higher preoperative disability demonstrated statistically significant greater improvements (compared to preoperative score) in parent-reported PedsQL at all timepoints after surgery, except at the 6-month&nbsp;parent-reported outcomes (p = 0.133).</p> <p><strong>Conclusion:</strong> Patients with worse preoperative disability, as assessed by PedsQL, experienced lower absolute PedsQL scores at all timepoints after surgery but had greater improvement in short- and intermediate-term PROs. We conclude that PedsQL is an efficient and accurate tool that can quickly assess patient disability in the preoperative period and predict both short-term and long-term surgical outcomes.</p> Shawyon Baygani, BS, Kristin Zieles,BS, Andrew Jea, MD, MHA Copyright (c) 2019 Shawyon Baygani, BS, Kristin Zieles,BS, Andrew Jea, MD, MHA https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23419 Tue, 08 Oct 2019 10:26:18 -0400 Screening transposon-mutant library of methicillin-resistant Staphylococcus aureus for mutants with altered sensitivity toward three antimicrobial agents http://journals.iupui.edu/index.php/IMPRS/article/view/23406 <p><strong>Background and Hypothesis:</strong> Methicillin-resistant Staphylococcus aureus (MRSA) is resistant to almost all beta-lactam antibiotics, including oxacillin. To combat MRSA infections, recently, new classes of drugs are developed, such as oxadiazole DR1 and ESP404. We hypothesize that, if we identify MRSA mutants with altered sensitivity toward the drugs, those mutants will help us understand the antimicrobial mechanisms of the drugs.</p> <p><strong>Experimental Design or Project Methods:</strong> The screening was carried out on the Nebraska library consisting of 1,920 non-redundant transposon mutants of USA300 strain in 96 well plates. First, the minimum inhibitory concentration (MIC) of each drug was determined. Then the transposon mutants were grown in tryptic soy broth (TSB) containing either 1/4× MIC or 4× MIC of the drugs for sensitive mutants and resistant mutants respectively. Due to the limited availability of ESP404, the drug was used to screen for sensitive mutants only.</p> <p><strong>Results:</strong> The first screening identified 81 mutants with altered drug sensitivity: 23 for oxadiazole DR1 (5 sensitive, 18 resistant); 20 for ESP404 (all sensitive); 38 for oxacillin (18 sensitive, 5 resistant).</p> <p><strong>Conclusion and Potential Impact:</strong> Screening of the transposon-mutant library identified multiple mutants that have altered sensitivity toward three drugs against S. aureus. Further verification of the phenotype and characterization of those mutants are expected to shed light on the antimicrobial mechanism of the drugs and open a door for the development of drug-potentiators.</p> Mahmoud Ahmed, George Beshara, Bryan Ubanwa, Taeok Bae Copyright (c) 2019 Mahmoud Ahmed, George Beshara, Bryan Ubanwa, Taeok Bae https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23406 Tue, 08 Oct 2019 10:26:44 -0400 Validation of a Postoperative Delirium Prediction Model http://journals.iupui.edu/index.php/IMPRS/article/view/23422 <p><strong>Background and Hypothesis:</strong> A prediction model utilizing the National Surgical Quality Improvement Program risk calculator for serious complications (NSQIPSC) and measures of cognitive function has been demonstrated to predict postoperative delirium incidence and severity in patients after non-cardiac surgery. The next step is to test this model’s generalizability by validating in different patient populations and evaluating the robustness of the model with and without parameters of cognition. We hypothesized that the prediction model would still function in predicting postoperative delirium in patients undergoing major thoracic surgery.</p> <p><strong>Experimental Design or Project Methods:</strong> A secondary data analysis of a randomized clinical trial involving 135 individuals who underwent major thoracic surgery from October 2013 to June 2015 was done. Delirium incidence and severity were tracked postoperatively through the Confusion Assessment Method for the ICU (CAM-ICU) and the Delirium Rating Scale-R-98 (DRS), respectively. The Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) was used to assess preoperative cognition. Linear regressions and specificity/sensitivity calculations were performed to evaluate the model.</p> <p><strong>Results:</strong> One hundred and thirty-two patients with a mean age of 59±13 years, 33/132 CAM-ICU+, and a mean DRS peak of 3.12±3.43 were analyzed. Linear regression of NSQIP-SC scores moderately predicted delirium severity (P&lt;0.001, Adjusted R2: 0.28). Addition of a cognitive measure did not significantly improve the model (P&lt;0.001, Adjusted R2: 0.29). NSQIP-SC moderately predicted delirium incidence (sensitivity: 60.61%, specificity: 68.69%, AUROC: 0.73). Addition of a cognitive measure (sensitivity: 61.54%, specificity: 68.69%, AUROC: 0.73) did not significantly improve the model. </p> <p><strong>Conclusion and Potential Impact:</strong> NSQIP-SC was a predictor variable for delirium incidence and severity. Addition of a cognitive measure did not have any appreciable effect on the prediction model. Further expanding the scope of the model can grant clinicians a tool to identify patients at risk for developing postoperative delirium.</p> Ashok Biju, B.S., Babar Khan, M.D. M.S., Heidi . Lindroth, Ph.D. R.N Copyright (c) 2019 Ashok Biju, B.S., Babar Khan, M.D. M.S., Heidi . Lindroth, Ph.D. R.N https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23422 Tue, 08 Oct 2019 10:27:10 -0400 Inhibition of Type 2 Sodium-Glucose Transporters and Na+/H+ Exchanger-1 produces similar cardioprotective effects in response to ischemiareperfusion injury http://journals.iupui.edu/index.php/IMPRS/article/view/23423 <p><strong>Background and Hypothesis:</strong> Recent studies indicate that inhibition of Type 2 Sodium-Glucose Transporters (SGLT2i) augments diastolic filling volume and mitigates myocardial ischemic injury. This study tested the hypothesis that inhibition of the Na+/H+ Exchanger-1 (NHE-1) mimics the cardioprotective effects of SGLT2i in response to ischemia-reperfusion injury.</p> <p><strong>Experimental Design or Project Methods:</strong> Lean swine (~50 kg) were anesthetized, a thoracotomy performed, and perivascular flow transducers placed around the left anterior descending (LAD) and circumflex coronary (LCX) arteries. A pressure-volume (PV) catheter was then inserted into the left ventricle. Swine received a 15 min infusion of vehicle (DMSO; n = 3), the SGLT2i Canagliflozin (30 µM; n = 3), or the NHE-1 inhibitor Cariporide (1µM; n = 3) prior to a 60 min total occlusion of the LCX and 2-hour reperfusion period. Following reperfusion, the LCX was re-occluded and a 2.5% Patent Blue 5 solution was administered to identify area at risk. The heart was excised, sectioned, and incubated in a 2,3,5-triphenyltetrazolium chloride (TTC) solution. Images were collected and analyzed for area at risk and infarct size. </p> <p><strong>Results:</strong> In the vehicle treated group, 2 of the 3 swine studied died prematurely before the completion of the protocol; one at baseline and one during ischemia. Our initial findings support that left ventricular end diastolic volume increases in response to regional myocardial ischemia in swine that received either Canagliflozin or Cariporide. This increase in diastolic volume was associated with an increase in stroke volume (i.e. Frank-Starling effect) and a reduction in myocardial infarct size in both treatment groups. Blood pressure tended to decrease to a similar extent in all groups. </p> <p><strong>Conclusion and Potential Impact:</strong> These pilot studies suggest that inhibition of SGLT2 and NHE-1 produces similar functional and protective effects in response to regional ischemia-reperfusion injury. Further experiments are needed to corroborate these findings and examine the extent to which SGLT2i directly modulates NHE-1 activity.</p> Bianca S. Blaettner, Hana E. Baker, Adam G. Goodwill, Hannah E. Clark, Michael C. Kozlowski, Johnathan D. Tune Copyright (c) 2019 Bianca S. Blaettner, Hana E. Baker, Adam G. Goodwill, Hannah E. Clark, Michael C. Kozlowski, Johnathan D. Tune https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23423 Tue, 08 Oct 2019 10:27:54 -0400 Alcohol Induced Oxidative Stress leads to Blood Brain Barrier Dysfunction in an IPSC-derived Model http://journals.iupui.edu/index.php/IMPRS/article/view/23424 <p><strong>Background:</strong> The NIH states that 6.2% of adults in the U.S. are diagnosed with an alcohol use disorder. Previous animal studies have indicated that ethanol can lead to dysfunction in the blood brain barrier (BBB). The BBB is critical in maintaining homeostasis between the vasculature and the brain parenchyma. The mechanisms underlying alcohol-induced barrier breakdown are relatively unclear. Utilizing a human derived BBB model we investigated the cellular effects of ethanol on several critical barrier properties. We hypothesized that human induced pluripotent stem cell (iPSC)-derived brain microvascular endothelial cells (BMECs) produced excessive reactive oxygen species (ROS) due to increased ethanol metabolism and ultimately contributes to a leaky BBB.</p> <p><strong>Designs/Methods:</strong> iPSC-derived BMECs were exposed to ethanol (10, 50, 100 mM) for one hour and barrier integrity was monitored with trans-endothelial electrical resistance (TEER) and sodium fluorescein permeability assays. A DCFDA assay kit monitored ROS. To determine the role of ethanol metabolism, BMECs were treated with 4-methylpyrazole (4-MP), an ethanol metabolism inhibitor, and Trolox, a ROS scavenger. Finally, we determined the role of alcohol-induced ROS on barrier integrity in the presence of 4-MP and/or Trolox.</p> <p><strong>Results:</strong> iPSC-derived BMECs exposed to ethanol (50 and 100 mM) had a significant reduction in TEER indicative of an impaired BBB. BMECs exposed to ethanol showed an increase in DCFDA fluorescence indicating an increase in ROS production. When BMECs were treated with 4-MP or Trolox, alcohol-induced ROS production was decreased.</p> <p><strong>Potential Impact:</strong> Recent studies indicate that dysregulation of the BBB could be associated with the pathogenesis of neurological and psychiatric disorders. With such a high prevalence of alcohol consumption in the U.S. a significant portion of the population could be at a higher risk of developing a neurodegenerative disease. Determination of the underlying cellular mechanisms responsible for alcohol-induced barrier damage could potentially contribute to therapeutic interventions.</p> Wesley Borman, Kameron Bell, Jason Hughes, Scott G. Canfield, PhD Copyright (c) 2019 Wesley Borman, Kameron Bell, Jason Hughes, Scott G. Canfield, PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23424 Tue, 08 Oct 2019 10:28:26 -0400 INF-β treatment to prolong t-PA treatment window in cerebral ischemia http://journals.iupui.edu/index.php/IMPRS/article/view/23426 <p><strong>Background and Hypothesis:</strong> Cerebral ischemia is the 2nd leading cause of death worldwide. The pathological hallmarks of cerebral ischemia are cell damage, degradation of the blood brain barrier (BBB), and inflammation followed by resident microglia activation, peripheral immune cell infiltration and subsequent secondary neurodegeneration. The first line therapy for ischemic stroke is the thrombolytic, tPA. However, only ten percent of patients are eligible for this treatment – primarily due to the risk of cerebral hemorrhage, secondary to BBB breakdown. Less than ten percent of individuals with acute cerebral ischemia are eligible for tPA. The objective of this study is to establish whether matrix metalloproteinase (MMP) activity, implicated in exacerbating the cerebral infarct volume seen with delayed tPA treatment, can be suppressed with Interferon-β (IFN-β) and thus extend the therapeutic window of tPA.</p> <p><strong>Experimental Design:</strong> We first investigated the therapeutic effect of IFN-β co-administered with t-PA in the mouse model of transient middle cerebral artery occlusion/reperfusion. Second, using immunoblotting technique we investigated the expression levels of MMPs in brain endothelial and microglial cells following various treatment combinations of TNFα and PGE2, t-PA, and INF-β.</p> <p><strong> Results:</strong> First, we demonstrated that IFN-β coadministered with tPA reduces the infarct size in ischemic brains. Second, we demonstrated in microglial cells that MMP-9 expression induced by TNFα, PGE2, and tPA can be suppressed by IFN-β treatment. Our experiments to demonstrate the expression levels of MMP-3 and MMP-9 in brain endothelial cells require further optimization.</p> <p><strong>Conclusion and Potential Impact:</strong> Overall these data indicate that IFN-β treatment is a viable therapeutic candidate to suppress the deleterious effects seen in delayed tPA treatment in the setting of acute cerebral ischemia. Furthermore, our preliminary data indicate that the molecular target of IFN-β, in this setting, belong to the MMP family of proteins.</p> Kristopher D. Bosi, BS, Jui-Hung Yen, PhD Copyright (c) 2019 Kristopher D. Bosi, BS, Jui-Hung Yen, PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23426 Tue, 08 Oct 2019 10:28:53 -0400 Strategies to Increase Time in Range in Children Wearing Continuous Glucose Monitors http://journals.iupui.edu/index.php/IMPRS/article/view/23427 <p><strong>Background and Hypothesis:</strong> Optimal management of Type 1 Diabetes (T1D) requires careful orchestration of insulin dosing with blood glucose values, food intake, activity levels, and concurrent illness. This is particularly burdensome for caretakers of very young children. Continuous glucose monitors (CGMs) provide useful real-time blood glucose information. Yet, data suggest that their use in young children can reduce severe hypoglycemia but does not consistently improve the time that blood glucose values are in an optimal range.</p> <p><strong>Project Methods:</strong> We first analyzed survey data from a study of CGM use in 143 children 2-8 years of age). We then developed a semi-structured qualitative interview, and queried parents of children using CGM under 9 years of age (mean age 5.2±1.6 years, A1c 7.8±0.9%).</p> <p><strong> Results:</strong> Recurrent themes identified included: CGMs and remote monitoring brought parents peace of mind and permitted more flexibility in care. Trend arrows indicating rapid glucose decreases induced “anxiety” and “panic” in caretakers, due to fear of hypoglycemia and immediate safety. Rising trend arrows induced frustration, but were less likely to change management decisions, due to lack of immediacy of risk of long-term complications. CGM data can be “overwhelming”, but third party apps and “experience” increase helpfulness.</p> <p><strong> Conclusion/Potential Impact:</strong> Our initial data suggest that behavioral/educational interventions to improve time in range must incorporate implementable strategies, potentially using apps, to encourage parents to respond to and reduce time spent in higher blood glucose ranges.</p> Will Bridgeman, Linda DiMeglio Copyright (c) 2019 Will Bridgeman, Linda DiMeglio https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23427 Tue, 08 Oct 2019 10:29:18 -0400 Cell cycle analysis of MDA-MB-157 and APC knockdown cells http://journals.iupui.edu/index.php/IMPRS/article/view/23429 <p><strong>Background and Hypothesis:</strong>&nbsp;A majority of sporadic breast cancers include deficits in the expression of Adenomatous Polyposis Coli (APC), a tumor suppressor. Deficits in APC are more common in patients with TNBC (triple negative breast cancer), which is also a cancer prone to chemotherapy resistance. The Prosperi lab previously found that APC knockdown cells (APCKD) were resistant to Paclitaxel (PTX). We hypothesize that APCKD cells are resistant to PTX treatment through avoidance of G2/M arrest. This summer, my goal was to investigate the mechanism by which PTX works to arrest the cell cycle at G2/M.&nbsp;</p> <p><br> <strong>Experimental Design or Project Methods:</strong></p> <p><span style="text-decoration: underline;">Cell Synchronization:</span> To synchronize MDA-MB-157 and APCKD cells, we first tried serum starvation for 24-72 hours. Second, we tried synchronizing the cells using a double thymidine block as described (Bostock, 1971). In either protocol, cells were then stained with Propidium Iodide (PI) and flow cytometry was performed. <br> <span style="text-decoration: underline;">Paclitaxel (PTX) treatment and cell cycle analysis:</span> MDA-MB-157 and APCKD cells were grown to confluency and then treated with 0.078µM PTX for 12, 24, and 48 hours. Cells were stained with PI and flow cytometry was performed.</p> <p><br> <strong>Results:</strong></p> <p><span style="text-decoration: underline;">Cell synchronization:</span> APCKD cells cells have an increased cell population in the G2/M phase than the parental cells after serum starvation. Importantly, APC knockdown cells are not impacted by serum starvation up to 72 hours. Additionally, a double thymidine block is insufficient to synchronize MDA-MB-157 and APCKD cells. A double thymidine block did shorten the S phase and move MDA-MB-157 and APCKD cells closer to G0-G1 arrest, but did not synchronize. <br> <span style="text-decoration: underline;">Paclitaxel (PTX) treatment and cell cycle analysis:</span> MDA-MB-157 and APCKD cells treated for longer intervals experienced more cell death and were further arrested in G2/M.</p> <p><br> <strong>Conclusion and Potential Impact:</strong> We learned that MDA-MB-157 and APCKD cannot be easily synchronized using serum starving or a double thymidine block. Future investigations will require alternative methods of synchronization or will proceed without synchronization. Furthermore, APCKD cells do not avoid G2/M arrest when treated with Paclitaxel, indicating a different mechanism of PTX resistance.</p> Jamie Brown, Emily Astarita, Camden Hoover, Jenifer R. Prosperi Copyright (c) 2019 Jamie Brown, Emily Astarita, Camden Hoover, Jenifer R. Prosperi https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23429 Tue, 08 Oct 2019 10:30:39 -0400 Skin Infections in Indiana Nursing Homes http://journals.iupui.edu/index.php/IMPRS/article/view/23430 <p><strong>Background and Hypothesis:</strong> OPTIMISTIC (Optimizing Patient Transfers, Impacting Medical Quality, &amp; Improving Symptoms: Transforming Institutional Care) is a CMS (Centers for Medicare and Medicaid Services) demonstration project that aims to improve nursing home care. CMS provides enhanced reimbursement to nursing homes in the project when they are able to treat six conditions in the facility rather than being transferred to a hospital, one of which being skin infections. The purpose of this study is to examine the diagnosis, treatments, and outcomes of skin infections of residents in nursing homes enrolled in OPTIMISTIC.</p> <p><br> <strong>Experimental Design or Project Methods:</strong> A chart review of 26 cases was conducted using data from nursing home medical records of nine OPTIMISTIC facilities. Cases were randomly selected from billing episodes between November 2018 and May 2019. Data was entered into an Excel extraction tool created for the project and analyzed.</p> <p><br> <strong>Results:</strong> Of the randomly selected skin infections, 46.2% were diagnosed as non-purulent cellulitis and 23.1% as purulent cellulitis. Other infections included abscesses, infected pressure injuries, and shingles. Doxycycline was the most common antibiotic prescribed, used in 42.3% of the total skin infections. For non-purulent cellulitis, cephalexin was the next most used antibiotic (41.7%). Twelve of the skin infections were recurrent, and the most common antibiotic used for those infections was doxycycline. Of the twenty-six cases, twenty-three were resolved in the facility, one was recertified for a longer length of time, one was transferred to the hospital, and one was transferred to hospice.</p> <p><br> <strong>Conclusion and Potential Impact:</strong> This project provides a unique opportunity to examine diagnosis and treatment patterns of skin infections in nursing homes. The results of this study will be used to bolster and augment future efforts at nursing home care improvement for patients diagnosed with skin infections in the facility.</p> Kayla Brown, Kathleen Unroe, MD, MHA, Jennifer Carnahan, MD, MPH, MA Copyright (c) 2019 Kayla Brown, Kathleen Unroe, MD, MHA, Jennifer Carnahan, MD, MPH, MA https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23430 Thu, 10 Oct 2019 09:56:45 -0400 Starting at the Roots: Using Human-Centered Design to Develop a Sex and Pregnancy Education Curriculum for Adolescents in Kenya http://journals.iupui.edu/index.php/IMPRS/article/view/23431 <p>The rate of adolescent pregnancy worldwide remains unacceptably high. Sixteen million adolescent girls between the ages of 15 and 19, and 2 million under the age of 15 become pregnant each year. Ninety-five percent of these births occur in low-income countries, with four times the rate of adolescent pregnancy in the poorest regions of the world compared to high-income countries. There has been a shift globally to focus on the sexual and reproductive health needs of adolescents, including adolescent pregnancy. With increased awareness of this need has come a renewed call for evidence-based provision of adolescentfocused sexual and reproductive health (ASRH) services, as well as programs to prevent pregnancy in this age group.</p> <p><br> “Human-centered design” methodology is emerging as an innovative, feasible, and effective participatory approach to program design and implementation in health care. The standard Human Centered Design format includes 5 steps: (1) Understand people’s experiences, challenges, and priorities; (2) Use existing knowledge and new research to define and clarify the problem; (3) Prompt creative thinking to design many different solutions; (4) Build and workshop prototypes of ideas to quickly learn how they can be improved; and (5) Deliver solutions that meet the needs of the target population.</p> <p><br> Through this process, design team members identified a need to “help adolescents by removing the uncertainty that surrounds information on pregnancy and treatment choices”. To meet this need, we utilized participatory program development to build an adolescent-specific sex and pregnancy education program at Moi Teaching and Referral Hospital in Eldoret, Kenya.</p> Sean Buehler, BSPH, Julie Thorne, MD, MPH Copyright (c) 2019 Sean Buehler, BSPH, Julie Thorne, MD, MPH https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23431 Tue, 08 Oct 2019 10:31:11 -0400 Asthma in Indiana – Using a Community Health Matrix to Determine Asthma Health Factors for Indiana Counties http://journals.iupui.edu/index.php/IMPRS/article/view/23433 <p><strong>Background and Hypothesis:</strong> Asthma and its appropriate treatment is a public health issue in Indiana that Indiana Joint Asthma Coalition (InJAC), a partnership within CTSI, is attempting to address. This is done through state-wide coalition building, which unifies efforts regarding asthma health and education and promotes interprofessional collaboration. Because time and resources are limited, InJAC must choose the areas that would benefit most from their focused work. A matrix was developed to establish the 10 counties with poorest asthma health and high vulnerability to social determinants to aid in this choice. We hypothesize that the 10 counties with the highest vulnerability to social determinants of health will have the worst asthma health.</p> <p><br> <strong>Project Methods:</strong> Asthma health outcomes, contributing asthma-related variables, and social determinants of health were identified in all 92 counties in Indiana. Counties were compared by composite z scores to determine the top 10 counties with the poorest health statistics for asthma and social determinants. In addition, qualitative data will be used to identify local health coalitions that have the capacity and desire to work with InJAC to improve asthma treatment. InJAC will begin sessions with these counties to determine if long-term, sustainable, health promotions are feasible.</p> <p><br> <strong>Results:</strong> The top 10 counites that were identified as having the poorest asthma health and factors were Lake, Grant, Madison, Marion, Huntington, Vanderburgh, Howard, La Porte, Blackford, and Noble. The top 10 counties with highest vulnerability to social determinants were Owen, Ripley, Daviess, La Grange, Fayette, Wayne, Elkhart, Newton, Switzerland, and Marion.</p> <p><br> <strong>Potential Impact:</strong> The data from this matrix will help direct InJAC to the areas of Indiana with the most need for asthma coalition efforts. This will be done through improvement on education, awareness, and quality of care based on the Indiana State Asthma Plan. </p> Mikayla Burrell, Rachael Casey, MPH, Dennis Savaiano, PhD Copyright (c) 2019 Mikayla Burrell, Rachael Casey, MPH, Dennis Savaiano, PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23433 Tue, 08 Oct 2019 10:31:36 -0400 Xenograft Bone-Derived Collagen Scaffold for Myelomeningocele Management http://journals.iupui.edu/index.php/IMPRS/article/view/23439 <p><strong>Background and Hypothesis:</strong> Spina bifida is a neural tube defect resulting from an incomplete closure of the caudal neuropore. The most debilitating form of spina bifida, myelomeningocele (MMC), can present with Chiari II malformation with concomitant hydrocephalus, bowel and bladder abnormalities, and impaired motor function of the lower limbs. The incidence rate of spina bifida is 3.4 per 10,000 live births reported within the US. On average, the US spends $1,176,000,000 annually on patient management and treatment. Advancements in existing treatment options, namely fetal surgery, can greatly decrease neurological injury and related costs, but at the risk of fetal and maternal complications. Various tissue engineering methods have been proposed including biodegradable and synthetic scaffolds, seeded with or without bioactive proteins and stem cells, sutured or glued to the defect, and administered fetoscopically or through open fetal surgery. However, no combination of these methods is fully biointegratable, watertight, provides complete coverage with adequate mechanical strength, and is able to be administered fetoscopically.</p> <p><strong>Experimental Design:</strong> This study utilizes bovine and porcine bone to create an organic, flexible collagen scaffold that can be seeded with bioactive proteins and attached with adhesive for successful coverage of MMC defects.</p> <p><strong>Conclusion and Potential Impact:</strong> The natural matrix may allow for quicker host cell integration and greater mechanical strength compared to existing models. This study will characterize the mechanical strength, permeability, and biointegration of the proposed management of spina bifida.</p> Sadid Khan, Natalie Campbell, Sam Fathizadeh, Sean Bucherl, Eric Nauman, Ph.D. Copyright (c) 2019 Sadid Khan, Natalie Campbell, Sam Fathizadeh, Sean Bucherl, Eric Nauman, Ph.D. https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23439 Tue, 08 Oct 2019 10:32:08 -0400 Mind the Gaps: Facilitators and Barriers to Behavioral Change Related to Infant Mortality Rate http://journals.iupui.edu/index.php/IMPRS/article/view/23440 <p><strong>Background and Hypothesis:</strong> Marion County suffers from stark racial disparities in infant mortality rate (IMR) with black babies experiencing an IMR of 11.6 and white babies 4.5, to culminate at a state average of 7.3. Since 2016, WeCare Indiana has worked to reduce the IMR in Central Indiana by connecting pregnant, post-partum, and childbearing age women with Community Health Workers (CHW) and community organizations that help with areas including but not limited to baby supplies, housing, and food insecurity. The goal of this research was to identify factors that promote changes in behaviors in WeCare clients as well as barriers to behavior modification in relation to the five pillars of the WeCare program: food insecurity, mental health, smoking, breastfeeding, and safe sleep.</p> <p><strong>Experimental Design or Project Methods:</strong> Qualitative analysis of unstructured CHW follow-up notes from 2016 to 2018 of 1775 WeCare clients from mostly 13 highest risk zip codes for IM in Central Indiana was completed using keyword searches and clustering narrative entries into themes associated with the apriori areas of behavioral change.</p> <p><strong>Results:</strong> Several key influencers of behavioral change were identified: (1) CHW dissemination of eligibility information about WIC, (2) CHW referral of clients to behavioral health resources, and (3) Pack n Play resources bundled with Safe Sleep class. Persistent barriers to positive behavioral change included: (1) lack of transportation to food resources, (2) unmanaged stress propagating substance use and smoking, (3) employment interfering with breastfeeding, and (4) lack of Safe Sleep education for all caretakers.</p> <p><strong>Conclusion and Potential Impact:</strong> Next steps include sharing qualitative findings with CHW to reinforce positive behavioral change as well as investigating solutions for the barriers that prevent change within the five pillars of <em>WeCare Indiana.</em> Data from this study will be used to help interpret the calculated behavioral change scores relative to baseline measures that were found from quantitative analysis.</p> Jessica Chiang, Sarah Roth, Wilma Griffin, Paige DeChant, Debra Litzelman Copyright (c) 2019 Jessica Chiang, Sarah Roth, Wilma Griffin, Paige DeChant, Debra Litzelman https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23440 Tue, 08 Oct 2019 10:32:36 -0400 Platelet Lyophilization and Cryopreservation: A Means of Generating a Standardized In-Vitro Human Blood Clot http://journals.iupui.edu/index.php/IMPRS/article/view/23441 <p><strong>Background and Hypothesis:</strong> Numerous models, both <em>in-vitro</em> and <em>in-vivo</em>, exist for the analysis of blood-clotting pharmacodynamics. However, reliance on animal models, fresh blood, or lack of a complete component profile translates to little standardization/reproducibility of clotting parameters. Given the above limitations, the goal of our study was to generate a reproducible, physiologic human blood clot through cryopreservation and lyophilization of platelets. We hypothesized that the platelets exposed to either lyophilization or cryopreservation would have a prolonged ability to generate reproducible clots; however, clotting functionality might exhibit a setvalue decrease resulting from storage procedures.</p> <p><strong>Experimental Design or Project Methods:</strong> Initial efforts focused on understanding the metabolic effects of storing platelets, plasma, and red blood cells (RBCs) at either 25°C or 4°C over a period of 25days. Glucose consumption was utilized as a proxy for metabolic activity and assessed using a glucose-hexokinase assay. Secondly, RBCs, platelets, and plasma were stored with the aim of maximizing functionality for longer periods: RBCs – 4°C+CPDA1 solution and platelets – cryopreserved (-80°C) or lyophilized. Functionality of stored blood components was assessed over a period of 1-week via Thromboelastographic (TEG) kinetic clotting readouts. Blood samples were provided by healthy volunteers (n=6).</p> <p><strong>Results:</strong> Utilization of glucose by RBCs and platelets was significantly increased during storage at 25°C versus 4°C, with RBCs maintaining very consistent glucose consumption rates compared to platelets. Lastly, by 1-week of storage, wholeblood is able to maintain consistent clotting better than samples utilizing cryopreserved platelets.</p> <p><strong>Conclusion and Potential Impact:</strong> Through our work, we were able to generate a reproducible synthetic human blood clot after 1-week of whole-blood storage as described by TEG parameters; a major stride in the ultimate goal of generating a model for pharmaceutical testing. Although not all parameters were preserved, we were able to maintain relative differences, ensuring consistent coagulability even with cryopreserved platelets. Further work will be conducted to understand the effects of greater storage durations.</p> Alexei Christodoulides, Nathan J. Alves, PhD Copyright (c) 2019 Alexei Christodoulides, Nathan J. Alves, PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23441 Tue, 08 Oct 2019 10:33:01 -0400 Obesity decreases the contribution of Kv channels to hypoxic coronary vasodilation http://journals.iupui.edu/index.php/IMPRS/article/view/23442 <p><strong>Background and Hypothesis:</strong> Our group previously demonstrated that reductions in the functional expression of voltage-dependent K<sup>+</sup> (K<sub>v</sub>) channels contribute to impaired metabolic control of coronary blood flow in the setting of obesity. This study tested the hypothesis that obesity diminishes the contribution of K<sub>v</sub> channels to coronary vasodilation in response to hypoxemia.</p> <p><strong>Experimental Design or Project Methods:</strong> Control lean (<em>n = 7</em>) and obese (<em>n = 5</em>) swine were anesthetized and the heart exposed via left lateral thoracotomy. Coronary blood flow was measured in response to hypoxemia, before and after inhibition of K<sub>v</sub> channels by 4-aminopyridine (4-AP; 0.3 mg/kg, iv), by a flow probe placed about the left anterior descending coronary artery. Hypoxemia was induced by progressive increases in the amount of nitrogen introduced into the ventilator. Arterial blood samples were obtained at each reduction in arterial oxygenation via a catheter placed in the femoral artery.</p> <p><strong>Results:</strong> Blood pressure decreased from ~88 ± 5 mmHg to ~68 ± 6 mmHg (P = 0.01) as arterial PO<sub>2</sub> was reduced below 50 mmHg in both lean and obese swine (P = 0.51). In lean swine, coronary flow progressively increased from ~0.6 to &gt;3.0 ml/min/g as arterial PO<sub>2</sub> was reduced. This response was decreased by ~40% in obese swine and by ~30% in lean swine treated with 4-AP. Administration of 4AP had no effect on coronary flow in obese swine.</p> <p><strong>Conclusion and Potential Impact:</strong> These data support that K<sub>v</sub> channels contribute to increases in coronary flow in response to hypoxemia in lean swine and that reductions in K<sub>v</sub> channel function contribute to impaired hypoxic coronary vasodilation in obese swine. We propose that therapeutic targeting of obesity associated pathways (angiotensin-aldosterone system) known to influence K<sup>+</sup> channel expression could improve coronary microvascular function and cardiovascular outcomes in subjects with obesity. Supported by R01 HL136386; T35 HL 110854.</p> Hannah E. Clark, Hana E. Baker, Adam G. Goodwill, Bianca S. Blaettner, Michael C. Kozlowski, Johnathan D. Tune Copyright (c) 2019 Hannah E. Clark, Hana E. Baker, Adam G. Goodwill, Bianca S. Blaettner, Michael C. Kozlowski, Johnathan D. Tune https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23442 Tue, 08 Oct 2019 10:33:30 -0400 Association Between Peripheral Eosinophil Counts and Infant Lung Function http://journals.iupui.edu/index.php/IMPRS/article/view/23443 <p>&nbsp;</p> <p><strong>Background and Hypothesis:</strong> 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.</p> <p><strong>Experimental Design or Project Methods:</strong> 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.</p> <p><strong>Results:</strong> 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 FEV<sub>0.5</sub> and eosinophil counts is 80 samples.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> Alexander Close BS, James Slaven MS, Sydney Ross MS, Kirsten Kloepfer MD MS Copyright (c) 2019 Alexander Close BS, James Slaven MS, Sydney Ross MS, Kirsten Kloepfer MD MS https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23443 Tue, 08 Oct 2019 10:33:55 -0400 Individualized exercise and the health of the AYA cancer survivor population http://journals.iupui.edu/index.php/IMPRS/article/view/23444 <p><strong>Background + Hypothesis</strong></p> <p>Adolescent and young adult cancer survivors (AYACS: ages 15-39) have an 84.5% five-year survival rate.<sup>(1)</sup> AYACS have a 10 times greater risk to develop cardiac disease compared to healthy peers.<sup>(2)</sup> This is in part due to their lower physical activity.<sup>(3)</sup> Structured exercise in adult cancer survivors improves strength, fatigue, VO2, and antioxidant levels and it decreases markers of cellular damage<sup>(4, 5).</sup> AYACS could benefit similarly, reducing long-term health effects. Although evidence suggests exercise is beneficial in older cancer survivors, this has not been demonstrated in AYACS. We hypothesized that a 12-week one-on-one multi-modal, community-based exercise program would improve AYACS outcomes compared to baseline or inactive AYACS. The current study hopes to demonstrate the feasability of an exercise intervention in a community setting within Indianapolis.</p> <p><strong>Methods</strong></p> <p>Six individuals were included in a feasibility trial for a larger pilot study of 374 participants. On day 1, baseline assessments were performed for experimental outcomes: body composition, strength, flexibility, VO2peak, balance, plasma biomarkers, PA, psychological health, health-related quality of life, and fatigue. Mini reassessments were performed at week 5, measuring strength and VO2peak with an estimated 1-rep maximum and 6-minute Walk Test respectively; in the larger pilot study participants will be reassessed at weeks 12 and 24. Participants train for 60 minutes (20 cardio, 30 weights, 10 stretching) 3 times a week for 12 weeks, one-on-one with a cancer exercise specialist.</p> <p><strong>Results</strong></p> <p>The average change in VO2peak was +25.3% and in strength was +17.5% (no statistical analysis). Adherence was 90.9%.</p> <p><strong>Conclusion + Potential impact</strong></p> <p>This trial suggests the feasibility of a pilot larger study. The greatest limitation was that the population sample was not within the AYACS age range. However, as the goal was to show feasibility rather than to prove efficacy, the sample gave useful information.</p> <p><strong>Sources</strong> (1-5) <br>1. Anderson C, Smitherman AB, Nichols HB. Conditional relative survival among long-term survivors of adolescent and young adult cancers. Cancer. 2018;124(14):3037-43. 2. Armstrong GT, Kawashima T, Leisenring W, Stratton K, Stovall M, Hudson MM, et al. Aging and risk of severe, disabling, life-threatening, and fatal events in the childhood cancer survivor study. J Clin Oncol. 2014;32(12):1218-27. 3. Wu YP, Yi J, McClellan J, Kim J, Tian T, Grahmann B, et al. Barriers and Facilitators of Healthy Diet and Exercise Among Adolescent and Young Adult Cancer Survivors: Implications for Behavioral Interventions. J Adolesc Young Adult Oncol. 2015;4(4):184-91. 4. Repka CP, Hayward R. Oxidative Stress and Fitness Changes in Cancer Patients after Exercise Training. Med Sci Sports Exerc. 2016;48(4):607-14. 5. Repka CP, Hayward R. Effects of an Exercise Intervention on Cancer-Related Fatigue and Its Relationship to Markers of Oxidative Stress. Integr Cancer Ther. 2018;17(2):503-10. </p> Adam Corya, Jacob Conroy, Abigail Bolt, Jessica Ricks, Nicholas Kelly, Danielle Halsey, Tammy Sajdyk, Jamie Renbarger Copyright (c) 2019 Adam Corya, Jacob Conroy, Abigail Bolt, Jessica Ricks, Nicholas Kelly, Danielle Halsey, Tammy Sajdyk, Jamie Renbarger https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23444 Thu, 10 Oct 2019 09:58:39 -0400 Nef Alleles derived from HIV-Positive Patients with Pulmonary Hypertension Affect Production and Function of Exosomes in Endothelial Colony Forming Cells http://journals.iupui.edu/index.php/IMPRS/article/view/23447 <p><strong>Background:</strong> The HIV positive population has a &gt; 300 fold higher risk for developing pulmonary hypertension (PH) as compared to the general population. PH is a disease characterized by elevated blood pressure in the pulmonary vasculature and is currently with no cure. Experiments with primates found that Nef, an HIV encoded protein secreted in exosomes from HIV+ cells, plays an important role in the development of HIV-PH. HIV-Nef+ exosomes induce endothelial dysfunction, including premature senescence of endothelial colony forming cells (ECFCs). We hypothesize that exosome functions and concentrations in HIV+ patients may vary based on Nef-mutations, and thus potentially define a predictor for the risk of developing PH.</p> <p><strong>Methods:</strong> Nef exosomes were isolated from HEK293? cells transfected with molecular constructs containing HIV Nef signature sequences recovered from patients with HIV-PH. The Nef exosomes were then used to treat ECFC. We measured cell proliferation (by MTT assay), apoptosis (cleaved caspase-3 by Western blot), and cell senescence (p16<sup>ink4A</sup> by flow cytometry) in ECFCs treated with Nef exosomes. LM-10-Nanosight was used to quantify exosome concentrations in patient plasma.</p> <p><strong>Results:</strong> ECFC treated with Nef exosomes induced a 30% reduction in cell proliferation (p&lt;0.05, MTT). Western blot analysis demonstrated no significant change in apoptotic signaling, and flow cytometric analysis of p16<sup>ink4A</sup> showed increased senescence in Nef exosome-treated ECFCs versus mock control. “Nef PH-transfected cells showed that decreased proliferation was dependent on specific mutations of Nef.</p> <p><strong>Conclusion:</strong> Expression of Nef in cells results in increased exosome production and treatment of ECFC with these exosomes resulted in decreased proliferation, which inducting the cells towards senescence based on signature sequences in Nef.</p> Arthur Cross-Najafi, Tyler Colbert, Sarvesh Chelvanambi, Sharilyn Almodovar, Sonia Flores, Matthias Clauss Copyright (c) 2019 Arthur Cross-Najafi, Tyler Colbert, Sarvesh Chelvanambi, Sharilyn Almodovar, Sonia Flores, Matthias Clauss https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23447 Thu, 10 Oct 2019 10:00:18 -0400 Defining the expression and function of COQ8B in TAA http://journals.iupui.edu/index.php/IMPRS/article/view/23449 <p><strong>Background and Hypothesis:</strong> Thoracic aortic aneurysm (TAA) is characterized by progressive enlargement of the aorta and predisposes to life-threatening aortic dissection. Even among patients with monogenic causes of TAA, variable expressivity and incomplete penetrance hinders clinical decisions. Recent studies identify the gene COQ8B, a key mediator of mitochondrial ubiquinone biosynthesis, as a candidate genetic modifier in TAA. First, we sought to describe the distribution and morphological appearance of mitochondria in aortic smooth muscle cells (SMCs). We further hypothesized that decreased expression of <em>COQ8B</em> alters the contractile homeostasis of SMCs and predisposes to SMC apoptosis.</p> <p><strong>Experimental Design or Project Methods:</strong> We cultured SMCs directly from aortic tissues acquired at the time of aortic replacement surgery or heart transplantation. Mitochondria were labeled in live and fixed SMCs using the MitoTracker dye. We knocked down expression of <em>COQ8B</em> in SMCs using siRNA (siCOQ8B). The effects of siCOQ8B were compared with SMCs treated with a non-targeting negative control siRNA (siNeg). RNA expression was measured using quantitative polymerase chain reaction (qPCR) at 48 to 72 hours after siRNA transfection. Apoptosis was measured using a TUNEL-based fluorescent assay.</p> <p><strong>Results:</strong> Mitochondria in aortic SMCs showed a predominantly perinuclear distribution along with the appearance of a network of thin, filamentous extensions outward from the nucleus. Compared with siNeg controls, siCOQ8B decreased COQ8B RNA levels by approximately 90% on average across 5 knockdown experiments (80 to 95% knockdown). siCOQ8B led to increased RNA levels of <em>CNN1</em> across all experiments, ranging from 1.4 to 7.4-fold increases. Meanwhile, siCOQ8b led to lower levels of <em>COL1A1</em> in 4 of 5 experiments, ranging from 1.1 to 1.9-fold decreases. siCOQ8B led to decrease in the percentage of apoptotic cells in comparison with siNeg. In the presence of 1.6mM of H<sub>2</sub>O<sub>2</sub>, there was a 15% increase in apoptosis in cells transfected with siCOQ8B, and a 4.5% increase in cells not treated with H<sub>2</sub>O<sub>2</sub>.</p> <p><strong>Conclusion and Potential Impact:</strong> Decreased expression of COQ8B via siRNA knockdown alters the expression of genes important for SMC homeostasis, causing the SMC to be more susceptible to oxidative stress and inducing apoptosis. Future studies will further explore the <br>role of COQ8B on SMC contractility, matrix production, and apoptosis and elucidate the underlying mechanism. Ultimately, this insight may lead to novel clinical approaches and therapies for patients with TAA.</p> Christian C. Cuevas, Stephanie M. Ware, Benjamin J. Landis Copyright (c) 2019 Christian C. Cuevas, Stephanie M. Ware, Benjamin J. Landis https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23449 Tue, 08 Oct 2019 10:34:31 -0400 Prior Anatomy and Physiology – Benefits for Medical Students http://journals.iupui.edu/index.php/IMPRS/article/view/23451 <p><strong>Background and Hypothesis:</strong> Recent research in anatomy education found no relationship between prior anatomy coursework and gross anatomy outcomes at the professional school level (Robertson et al., 2019). Here we investigated whether prior coursework in anatomy and/or physiology produced differences in Human Structure course outcomes, study habits, or change thereof among first year medical students.</p> <p><strong>Project Methods:</strong> A survey was administered at the beginning and end of Human Structure for the 2016-2017 and 2017-2018 classes, collecting data about planned study habits, reported study habits, course outcomes, prior coursework, and other demographic information. The survey used Likert scale and open-ended questions. The data were then analyzed using SPSS; independent samples t-tests were used to compare final Human Structure grades, specific types of study methods and changes thereof, and overall study habit change among students with and without prior anatomy and/or physiology coursework.</p> <p><strong>Results:</strong> It was found that students with prior coursework deviated less from initial study plans, employed certain methods more, and outperformed students without such coursework. Compared to their counterparts, students with prior anatomy or physiology at the college level or higher changed responses to 6.43% fewer questions regarding study habits between the start and end of Human Structure (p=0.015). These students with prior experience used comparatively more text-based, lab-based, and web-based resources, also showing decreased change in the aforementioned categories as well as in self-made study resources. Lastly, students who took separate anatomy and physiology classes at a college level or higher scored 3.72% higher in Human Structure at IU (p=0.029).</p> <p><strong>Conclusion and Potential Impact:</strong> Students without prior coursework in anatomy and physiology demonstrated relative uncertainty in their study habits during Human Structure in medical school. It may be productive to offer such students extra help in devising a study plan at the start of the course.</p> Lindsey d’Arnaud, Polly R. Husmann, PhD Copyright (c) 2019 Lindsey d’Arnaud, Polly R. Husmann, PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23451 Tue, 08 Oct 2019 10:35:31 -0400 Perception and Outcome of PEM Fellows following POCUS Curriculum Implementation http://journals.iupui.edu/index.php/IMPRS/article/view/23453 <p><strong>Introduction:</strong> Point-of-care ultrasonography (POCUS) in the emergency department provides the clinician with a real-time, non-invasive and rapid assessment of their patients. Currently, there is a lack of published POCUS curriculum guidelines for pediatric emergency medicine (PEM) trainees. In this regard, we developed a POCUS curriculum for fellows at a newly established PEM fellowship at our institution to understand their perception and its effectiveness.</p> <p><strong>Methods:</strong> We enrolled all current PEM fellows (n = 4) who had limited to no prior experience with POCUS to participate in program implementation in January 2018. Their progress was followed over time until graduation. The curriculum included didactics hands-on training in the presence (supervised) and absence (solo) of an expert instructor. In addition to the curriculum, the fellows were also required to complete a pre- and post-survey assessment.</p> <p><br> <strong>Results:</strong> Before curriculum implementation, barriers of POCUS common to all fellows (100%) included image interpretation and comfort level. Conversely, this improved with only 1 fellow (25%) suggesting these barriers. Interestingly, nearly all fellows (75%) found POCUS to be time-consuming following the curriculum implementation. Knowledge-base assessment showed improvement from an average score of 50% pre-test versus 79% post-test. The number of scans increased with time, but qualitative measures of image acquisition had a minimal improvement on a self-assessment quality assurance (QA) Likert scale. Average solo QA rating by an expert was higher (Mean 3.23 + SEM 0.15) than the average by the fellows' selfassessment (3.07 + 0.18). Overall sensitivity of POCUS examinations was 90.32% (61.92%93.07%) and specificity was 99.11% (94.07%-99.57%).</p> <p><strong>Conclusion:</strong> POCUS curriculum implementation can improve the PEM fellow's skills while enhancing their experience. While there are challenges for the utilization of POCUS, this study shows that the implementation of such a curriculum may have a positive effect on skill, knowledge development as well as perception.</p> Steven Dawson, Ben Nti Copyright (c) 2019 Steven Dawson, Ben Nti https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23453 Tue, 08 Oct 2019 10:35:57 -0400 Attitudes towards nutrition education among pediatricians and guardians http://journals.iupui.edu/index.php/IMPRS/article/view/23454 <p><strong>Background and Hypothesis:</strong> Childhood obesity rates in the United States are at historic highs. In Lake County, Indiana, the obesity rates of WIC children ages 2-5 years old is 12.1%. Obesity management is left in the hands of practitioners’ clinical judgment; yet, a survey found that the average medical school devotes less than 20 hours to nutrition education. Since 62% of patients believe that their physicians can help them lose weight, having physicians who are not educated on nutrition leaves patients without the help they need. <br><br>Surveys were developed to assess the level of nutrition counseling provided by physicians and how patients/guardians prefer to be educated. Our hypothesis is that pediatricians will benefit from further nutrition education and that guardians will desire an accessible online source.</p> <p><strong>Methods:</strong> A physician survey was composed to determine how much nutrition education pediatricians received in medical school/residency and how they currently educate their patients. The goal of the guardian survey was to determine who in the household feeds the children and how they would prefer to receive education.</p> <p><strong>Results:</strong> The surveys will be utilized for future research, and the results will help determine the approach for educating physicians and guardians. A booklet of healthy recipes was also developed to educate on healthy eating and as a participation benefit. The goal of the booklet was to choose easy, child friendly recipes that the family could cook together. To gain background on nutrition education, we observed the different education methods of local pediatricians and reviewed the literature.</p> <p><strong>Potential Impact:</strong> Intervention at both the clinical and community levels will be important for improving long-term health outcomes in pediatric patients. The knowledge gained from these surveys will aide in the development of programs needed to provide physicians, guardians, and patients with proper nutrition education.</p> Madison DeGoey, Erica Kubascik, Dr. Uzelac, Dr. Simpson, Dr. Kostrominova Copyright (c) 2019 Madison DeGoey, Erica Kubascik, Dr. Uzelac, Dr. Simpson, Dr. Kostrominova https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23454 Tue, 08 Oct 2019 10:36:33 -0400 Body Morphometrics in NAFLD Population http://journals.iupui.edu/index.php/IMPRS/article/view/23455 <p><strong>Introduction:</strong> 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. </p> <p><strong>Methods:</strong> 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.</p> <p><strong>Results:</strong> 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.</p> <p><strong>Conclusion:</strong> 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.</p> Harkeerat Dhami, Niharika Samala Copyright (c) 2019 Harkeerat Dhami, Niharika Samala https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23455 Tue, 08 Oct 2019 10:36:59 -0400 Evaluation of Treatment for Volumetric Muscle Loss in a Porcine Tibia Segmental Bone Defect Model http://journals.iupui.edu/index.php/IMPRS/article/view/23456 <p><strong>Background and Hypothesis:</strong> Injury to soft tissue with volumetric muscle loss (VML) concomitant to segmental bone defects (SBDs) can be difficult to treat. One potential option is skeletal muscle autograft (SMA). We hypothesize the maximal torque measured <em>in vivo</em> and Radiographic Union Scale for Tibial Fractures (RUST) scores will be increased in the VML+SBD+SMA group compared to the VML+SBD group, but will be decreased compared to SBD alone.</p> <p><strong>Experimental Design or Project Methods:</strong> 18 male, Yucatan minipigs aged 1821 months were randomized into 3 groups: VML+SBD+SMA, VML+SBD, and SBD. This study is still ongoing with 12 pigs having had the surgery performed to date. <em>In vivo</em> muscle testing was performed prior to surgery in both hindlimbs to assess baseline strength. RUST scores and <em>in vivo</em> muscle testing was/will be performed at 1, 2, and 3 months post-injury. All procedures were conducted following approved IACUC protocol.</p> <p><strong>Results:</strong> Muscle testing results demonstrate no significant difference between control and operative limbs pre-surgery (T-tests, p= 0.673, 0.824, and 0.739 for VML+SBD+SMA, VML+SBD and SBD groups, respectively), or between the 3 different groups pre- or post-surgery (2-way ANOVA, p=0.788). The average RUST score for each treatment group at 2 months was 6.8±2.2, 7.7 7.7±1.1, and 8.6 8.6±0.4 for VML+SBD+SMA (n=2), VML+SBD (n=5), and SBD (n=3) groups, respectively.</p> <p><strong>Conclusion and Potential Impact:</strong> The lack of difference amongst groups on pre-operative muscle testing and a difference post-operatively between control and operative legs post-operatively help to validate this study. Preliminary results demonstrate a reduced RUST score for the VML+SBD+SMA group; however, this along with the lack of difference found in muscle testing results postoperatively may be an artifact due to small sample size and/or timing. Development of a model for VML and SBD will allow for testing of therapies for treat clinical problems.</p> Nicklaus Diggins, Seungyup Sun, Benjamin T. Corona, Aamir Tucker, Alex Brinker, Michael Savaglio, Gremah Adam, Zachary Gunderson, Roman M. Natoli, Todd O. McKinley, Melissa A. Kacena Copyright (c) 2019 Nicklaus Diggins, Seungyup Sun, Benjamin T. Corona, Aamir Tucker, Alex Brinker, Michael Savaglio, Gremah Adam, Zachary Gunderson, Roman M. Natoli, Todd O. McKinley, Melissa A. Kacena https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23456 Tue, 08 Oct 2019 10:37:19 -0400 Neonatal Follow Up Study: A Retrospective Chart Review of Developmental Outcomes http://journals.iupui.edu/index.php/IMPRS/article/view/23458 <p><strong>Background and Hypothesis:</strong> According to the CDC, Preterm birth is defined as a baby born before 37 weeks of gestation.&nbsp;<sup>1</sup> In 2016 preterm birth affected 1 in every 10 infant born in the United States.<sup>1</sup> 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.<sup>3</sup> We believe that with the NFC, we will be able to develop a standard for preterm babies and the normalization of their development.</p> <p><strong>Experimental Design or Project Methods:</strong> 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.</p> <p><strong>Results:</strong> 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.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> <p>1) Preterm Birth https://www.cdc.gov/reproductivehealth/MaternalInfantHealth/PretermBirth.htm 2) Suave, R., &amp; 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</p> Heather Wolfe, MD, Denise Gilham, NP, Susan Stace, RN, CPN, Uchechukwu Emili, BS Copyright (c) 2019 Heather Wolfe, MD, Denise Gilham, NP, Susan Stace, RN, CPN, Uchechukwu Emili, BS https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23458 Tue, 08 Oct 2019 10:37:52 -0400 Buprenorphine Metabolism in Human Liver, Human Placenta, and Recombinant Cyp19 Microsomes http://journals.iupui.edu/index.php/IMPRS/article/view/23459 <p><strong>Background and Hypothesis:</strong> <br> Buprenorphine (BUP), a partial opioid agonist, is commonly used to treat opioid use disorder in pregnant women. BUP is metabolized in the liver by cytochrome P450s (CYP) and UDPGlucuronosyltransferases (UGT). Compared to non-pregnant women, higher doses of BUP are needed during pregnancy to maintain therapeutic concentrations. We hypothesize that the placenta has a role in the metabolism of BUP, thus contributing to the need for higher doses during pregnancy.</p> <p><strong>Experimental Design or Project Methods: :</strong> BUP was incubated with human liver microsome (HLM), human placenta microsome (HPM) or recombinant CYP19 microsome (rCYP19). Norbuprenorphine (Nor-BUP) formation was measured by HPLC/MS/MS (Thermo TSQ Quantum Ultra). Positive controls included midazolam for HLMs and testosterone for HPMs and rCYP19, and negative controls were without NADPH. Initial experiments assessed linearity with time and protein concentration. Subsequently, BUP concentrations were varied and Nor-BUP formation vs. BUP concentration was fitted to the Michaelis–Menten equation (GraphPad Prism) to calculate <em>K<sub>m</sub></em> and V<sub>max</sub>.</p> <p><strong>Results:</strong> The <em>K<sub>m</sub></em> was 11.89 μM and V<sub>max</sub> was 0.4627 μmol/min/mg for Nor-BUP formation following incubation of BUP (0-20 µM) with HLM. Higher concentrations of BUP indicated non-typical Michaelis–Menten kinetics. Results from the HPM show metabolism of BUP to Nor-BUP is present in the placenta. Incubation of BUP in rCYP19, an enzyme known to be in human placenta, resulted in Nor-BUP formation, indicating its role in BUP metabolism.</p> <p><strong>Conclusions and Potential Impact: :</strong> The<em> K<sub>m</sub></em> and V<sub>max</sub> values generated from concentrations of 5-20 μM in HLM is comparable with existing data. Because multiple enzymes breakdown BUP with different <em>K<sub>m</sub></em> and V<sub>max</sub> values, a curve not adhering to the normal Michaelis–Menten shape over 20 μM BUP is reasonable in HLMs. Formation of Nor-BUP following incubation of BUP in rCYP19 and HPMs suggest their role in the metabolism of BUP in pregnant women. Understanding the role of placenta in metabolizing BUP will provide insight to fetal drug exposure. Placental metabolism of BUP would possibly indicate lower exposure to BUP but higher exposure to nor-BUP to the fetus. Future directions of this study will quantify placental metabolism of BUP, investigate other pathways of BUP metabolism in the placenta, and determine drug interactions that may exacerbate Neonatal Abstinence Syndrome when coadministered with BUP.</p> Kaitlyn Enderle, Xiaomei Zheng, Sara Quinney, PharmD, PhD Copyright (c) 2019 Kaitlyn Enderle, Xiaomei Zheng, Sara Quinney, PharmD, PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23459 Tue, 08 Oct 2019 10:38:16 -0400 “Women Never Use Drugs Alone”: Assessing Stigma & Access to Care among Women who Use Drugs. http://journals.iupui.edu/index.php/IMPRS/article/view/23460 <p><strong>Background</strong></p> <p>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.</p> <p><strong>Methods</strong></p> <p>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.</p> <p><strong>Results</strong></p> <p>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.</p> <p><strong>Conclusions and Recommendations </strong></p> <p>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.</p> Amanda Essex, Carrie Lawrence, Ph.D. CFLE CHES, Brooklyn Sean Turner Copyright (c) 2019 Amanda Essex, Carrie Lawrence, Ph.D. CFLE CHES, Brooklyn Sean Turner https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23460 Tue, 08 Oct 2019 10:40:22 -0400 Patellar Tilt in Two Modern Total Knee Arthroplasty Systems: A Matched-Cohort Analysis http://journals.iupui.edu/index.php/IMPRS/article/view/23461 <p><strong>Background and Hypothesis:</strong> 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.</p> <p><strong>Experimental Design or Project Methods:</strong> 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. </p> <p><strong>Results:</strong> There were no differences between study cohorts for demographics, covariates, or preoperative patellar tilt (<em>p</em>≥0.479). Asymmetric lateralized trochlear groove TKAs had significantly less postoperative patellar tilt compared with symmetric trochlear groove TKAs (<em>p</em>&lt;0.001). Patellar tilt and femoral implant type had no correlation to pain scores (Knee Society), UCLA Activity Level, or satisfaction (<em>p</em>≥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 (<em>p</em>=0.033).</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> Braeden W. Estes, BS, Lauren Pitz, BS, Evan R. Deckard, BSE, R. Michael Meneghini, MD Copyright (c) 2019 Braeden W. Estes, BS, Lauren Pitz, BS, Evan R. Deckard, BSE, R. Michael Meneghini, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23461 Tue, 08 Oct 2019 10:41:00 -0400 INVESTIGATING A NOVEL MODIFIABLE FACTOR AFFECTING RENAL FUNCTION AFTER PARTIAL NEPHRECTOMY: CORTICAL RENORRHAPHY http://journals.iupui.edu/index.php/IMPRS/article/view/23462 <p><strong>Background:</strong> Partial nephrectomy is a common treatment for the removal of renal masses. Typically, during the procedure, stitches are used to close two layers of the kidney—both deep and superficial. Renorrhaphy of the superficial layer, thought to reduce the risk of bleeding and urine leak, is routinely performed but has not been sufficiently studied. </p> <p><strong>Hypothesis:</strong> Cortical renorrhaphy is a modifiable factor affecting renal function after partial nephrectomy. Omitting this step will preserve renal parenchyma without significantly increasing complications.</p> <p><strong>Methods:</strong> A randomized, controlled trial is underway. Interim statistical analysis has been performed on the data being collected. Patients underwent partial nephrectomy with or without cortical renorrhaphy according to their randomized group assignment. Three-dimensional models were constructed using semi-automatic segmentation planimetry of the kidney prior to surgery and at 4-months after tumor resection to determine volume loss in the operated kidney.</p> <p><strong>Results:</strong> The median (range) volume loss in the non-renorrhaphy group (n=8), 13% (0-24%), was trending lower than the renorrhaphy group (n=8), 22% (12-39). Using multiple linear regression, experimental group (p=0.0808) and warm-ischemia time (p=0.0995) were significant at the 0.1 level. Tumor size was not statistically significant (p=0.2644). There was one Clavien 3 complication in each group: The renorrhaphy group had one urine leak requiring a drain, and the non-renorrhaphy group had one postoperative bleed requiring selective embolization. Demographics were comparable among the two groups with both having 4 white males and 4 white females. The mean age (58 and 55 for renorrhaphy and non-renorrhaphy, respectively) and tumor size were also comparable.</p> <p><br> <strong>Conclusion and impact:</strong> A trend of increased volume loss from cortical renorrhaphy is seen as predicted by retrospective data. Completion of the trial is needed to conclude whether this is statistically significant.</p> Anna Fenner, Clint Bahler, MD Copyright (c) 2019 Anna Fenner, Clint Bahler, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23462 Tue, 08 Oct 2019 10:41:41 -0400 Gastroschisis: Predicting outcomes through prenatal imaging and the current state of care in a single institution http://journals.iupui.edu/index.php/IMPRS/article/view/23465 <p><strong>Background:</strong> Gastroschisis occurs in 1 out of 2,000 births with survival rates partially contingent on intestinal complications and time to establishing feeding. Enhancements in prenatal imaging have given better insight into postnatal outcomes. The goal of this study is to examine the gastroschisis patient population and changes to care at Riley Hospital for Children in the modern era from 2010 to 2018. Additionally, we aim to utilize prenatal imaging to develop new prenatal prognostic indicators.</p> <p><strong>Methods:</strong> We performed a retrospective review of gastroschisis patients at Riley Hospital for Children from 2010 through 2018. We recorded demographics, prenatal data and imaging, early postnatal data, operative data, and patient outcomes and compared these variables to historic data gathered from studies at the same institution from the 2003-2009. Prenatal ultrasound factors were evaluated as a tool to predict the status of the bowel at birth. Statistical analyses were performed with chi square, univariate, and multivariate analyses.</p> <p><strong>Results:</strong> 134 patients were included in the study: complex (21), non-complex (113). Compared to non-complex gastroschisis, complex patients required longer median days to feeding initiation (46 vs 10, p&lt;0.001), full feeding (77.5 vs 23, p&lt;0.001), length of stay (LOS) (83 vs 33, p&lt;0.001), and TPN at discharge (p=0.031). Prenatal ultrasound factors showed a significant relationship between increased bowel dilation and non-complex gastroschisis (p=0.015). Compared to 2003-2009, 2010-2018 data showed a significant decrease in mortality (p=0.002) and use of primary closure (p=0.050). Multivariable logistic regression revealed complex gastroschisis to be predictive of total intubation days in the first 30 days of life, LOS, time to initial feeds, and time to full feeding (p&lt;0.001).</p> <p><strong>Conclusion and Potential Impact:</strong> An increase in bowel dilation on prenatal ultrasound can be used to predict non-complex gastroschisis at birth. Complex gastroschisis is associated with increased time to feeds, LOS, and intubation days.</p> Sarah G. Fisher, Cassandra M. Anderson, Nicole P. Steinhardt, Brandon P. Brown, MD, Brian W. Gray, MD Copyright (c) 2019 Sarah G. Fisher, Cassandra M. Anderson, Nicole P. Steinhardt, Brandon P. Brown, MD, Brian W. Gray, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23465 Tue, 08 Oct 2019 10:42:20 -0400 HMGB1 Receptors as Potential Novel Targets for Neuroblastoma http://journals.iupui.edu/index.php/IMPRS/article/view/23467 <p><strong>Background and Hypothesis:</strong> The pathogenesis of neuroblastoma remains elusive. In order to further understand the pathogenesis, it is necessary to identify molecular and cellular mechanisms of the cancer. One protein of interest is high-mobility group box 1 protein (HMGB1). HMGB1 has been implicated in cancer including tumor growth, invasion, angiogenesis, metastasis, relapse and therapeutic resistance. HMGB1 isoforms signals via different receptors including the chemokine receptor, CXCR4, receptor for advanced glycation end products (RAGE), and the prominent inflammation-associated receptor, toll-like receptor 4 (TLR4). To gain insight into the possible impact of HMGB1 on the SH-SY5Y neuroblastoma cell line, we determined the degree to which CXCR4, TLR4 and RAGE are present on SH-SY5Y cells. We hypothesize that the presence of these receptors in SH-SY5Y cells may mediate proliferation of neuroblastoma cells and other types of cancer.</p> <p><strong>Experimental Design or Project Methods:</strong> The SH-SY5Y cell line (ATCC® CRL-2266™) was derived from a metastatic bone tumor in a 4 year old female. Cells were lysed in lysis buffer, electrophoresed on a 10% SDS-PAGE, and blotted onto PVDF membrane. After blocking, the membranes were incubated with primary antibodies against the receptor protein overnight at 4 °C, and then with HRP-conjugated secondary antibodies for 1h. Protein bands were visualized with a SuperSignal West Pico Chemiluminescent Substrate.</p> <p><strong>Results: To be finalized </strong></p> <p><strong>Conclusion and Potential Impact:</strong> Investigating SH-SY5Y cell line for the presence of these inflammation-associated receptors could potentially serve as a model to better understand the role of these receptors in cancer research.</p> Christian Garcia, Jared Smith, Peter Malicky, Fletcher White Copyright (c) 2019 Christian Garcia, Jared Smith, Peter Malicky, Fletcher White https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23467 Tue, 08 Oct 2019 10:44:00 -0400 Polyethylene Bearing Wear in the Oxford Unicompartmental Knee http://journals.iupui.edu/index.php/IMPRS/article/view/23469 <p><strong>Background and Hypothesis:</strong> The Oxford Knee is a unicompartmental knee replacement (UKR) for patients with end-stage knee osteoarthritis (OA). It contains a polyethylene (PE) mobile bearing between the femoral and tibial components that helps maintain normal knee kinematics. Unfortunately bearing wear is one of the leading causes of UKR failure. In this study, we sought to investigate PE wear rate over time and the initial deformation (creep) in early months.</p> <p><strong>Project Methods:</strong> This study used radiostereometric analysis (RSA) to precisely track polyethylene bearing thickness over time in 40 medial Oxford UKR patients. The patients underwent stereo radiographs at 1 week post-op (reference), 3 months, 6 months, 1 year, 2 years, and 5 years. Model-based RSA software was used to determine the exact position of the femoral and tibial components, and Matlab was used to calculate the minimum linear distance between these two components, taken as bearing thickness.</p> <p><strong>Results:</strong> The average wear rate was 0.32mm/yr (SD 0.42) in the first 3 months, 0.16mm/yr (SD 0.55) from 3 to 6 months, and 0.07mm/yr (SD 0.03) between 1 and 5 years. No correlation was found between wear rate and BMI, fixation, implant size, or 5 year Oxford Knee Score.</p> <p><strong>Conclusion and Potential Impact:</strong> This was the first study to investigate detailed early and late time points of polyethylene wear in the Oxford UKR. Mean bearing wear was higher in the first 6 months, presumably due to creep, and significantly reduced from 1 year onwards, presumably while true wear took place.</p> Priyanka Ghosh, Hasan Mohammad, Stephen Mellon, David Murray Copyright (c) 2019 Priyanka Ghosh, Hasan Mohammad, Stephen Mellon, David Murray https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23469 Tue, 08 Oct 2019 10:44:56 -0400 Animal Assisted Interventions to Treat Pain and Associated Mental Health Comorbidities: A Systematized Review http://journals.iupui.edu/index.php/IMPRS/article/view/23493 <p><strong>Background and Hypothesis:</strong> Pain is cited as the primary reason patients access the healthcare system and is estimated to produce total financial cost of 500 billion dollars. However, pain can be difficult to treat, and is often associated with various mental health comorbidities, such as depression, anxiety, and PTSD, which can attenuate the effects of treatment. Thus, it is important to identify novel therapies that can both provide pain relief, and treat any associated mental health comorbidity. Animal Assisted Intervention (AAI) is one such possible treatment. Numerous studies have been published that highlight the potential of AAI in this domain. The aim of this study was to evaluate and synthesize the results of these studies.</p> <p><strong>Experimental Design or Project Methods:</strong> A literature search was conducted searching the databases MEDLINE, CINAHL, and EMBASE with search terms related to AAI, pain, anxiety, depression, and PTSD. 110 articles met the inclusion criteria and their results were reviewed.</p> <p><strong>Results:</strong> 24 articles analyzed outcomes related to pain, and 17 of these articles found that AAI significantly improved pain. Of these 17 articles, the average absolute magnitude of improvement in pain severity was 15%. 69 articles analyzed outcomes related to anxiety, and 37 found that AAI significantly improved anxiety. Of these 37 articles, the average absolute magnitude of improvement in anxiety symptom severity was 16%. 54 articles analyzed outcomes related to depression, and 29 found that AAI significantly improved depression. Of these 29 articles, the average absolute magnitude of improvement in depression symptom severity was 13%. 15 articles analyzed outcomes related to PTSD, and 11 found that AAI significantly improved PTSD symptoms. Of these 11 articles, the average absolute magnitude of improvement in PTSD symptom severity was 18%.</p> <p><strong>Conclusions and Potential Impact:</strong> AAI is promising as a complementary therapy for the treatment of pain as it is able to significantly improve pain while simultaneously treating common mental health comorbidities.</p> Jacob D. Graham, BS, Matthew J. Bair, MD, MS Copyright (c) 2019 Jacob D. Graham, BS, Matthew J. Bair, MD, MS https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23493 Thu, 10 Oct 2019 10:05:54 -0400 Comparison of Technical Outcomes in Instrumented Posterolateral Fusion (PLF) with and without Transforaminal Lumbar Interbody Fusion (TLIF) performed using Silicon Nitride cages http://journals.iupui.edu/index.php/IMPRS/article/view/23473 <p><strong>Background:</strong> 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 (Si<sub>3</sub>N<sub>4</sub>). Our hypothesis is TLIF with a Si<sub>3</sub>N<sub>4</sub> cage will have higher fusion rates, improved technical outcomes and fewer complications when compared to PLF alone. The Si<sub>3</sub>N<sub>4</sub> cage has advantageous surface properties compared to other interbody cages, promoting theoretically higher fusion rates for TLIF procedures.</p> <p><strong>Methods:</strong> 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).</p> <p><strong>Results:</strong> TLIF patients with a Si<sub>3</sub>N<sub>4</sub> 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=&lt;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).</p> <p><strong>Conclusion:</strong> Despite the difference in age between the procedure groups, TLIF with a Si<sub>3</sub>N<sub>4</sub> cage proved to be superior in fusion rates, lumbar lordosis, PI-LL mismatch, disc height and foraminal height restoration.</p> <p>&nbsp;</p> Mitchell Gray, B.A., Kyle Davis, B.S., Lauren Jagger, Micah Smith, M.D. Copyright (c) 2019 Mitchell Gray, B.A., Kyle Davis, B.S., Lauren Jagger, Micah Smith, M.D. https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23473 Tue, 08 Oct 2019 10:48:30 -0400 Gastric Per-Oral Endoscopic Myotomy (G-POEM) for the Treatment of Gastroparesis: A Single Center Experience http://journals.iupui.edu/index.php/IMPRS/article/view/23476 <p><strong>Background and Hypothesis:</strong> 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.</p> <p><strong>Experimental Design or Project Methods:</strong> 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.</p> <p><strong>Results:</strong> 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 &lt; 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 &lt; 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.</p> <p><strong>Conclusion and Potential Impact:</strong> G-POEM appears to be a safe and feasible treatment alternative for gastroparesis with significant short-term improvements in gastric emptying rates&nbsp;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.</p> Lennon Gregor, John Wo, John DeWitt, Anita Gupta, Brandon Yim, Martha Mendez, Mohammad Al-Haddad Copyright (c) 2019 Lennon Gregor, John Wo, John DeWitt, Anita Gupta, Brandon Yim, Martha Mendez, Mohammad Al-Haddad https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23476 Tue, 08 Oct 2019 10:48:54 -0400 Bag3 P209L myopathies and efficacy of blocking signaling pathways with the therapeutic peptide, MMI-0100 http://journals.iupui.edu/index.php/IMPRS/article/view/23478 <p><strong>Background and Hypothesis: </strong>The Bcl2-associated anthanogene (BAG) 3 protein is a member of the BAG family of cochaperones, which play a critical role in cellular processes, including protein degradation and turnover. Over 30 Bag3 mutations have been identified, including a Proline209Leucine (P209L) missense mutation which causes a severe childhood cardiomyopathy. The mechanism by which Bag3 mutations causes cardiomyopathy is currently unknown, but the p38/MAPK signaling cascade has been shown to be altered in our animal model that coexpresses the human Bag3 P209L gene. We hypothesized the cell-permeant peptide, MMI-0100, which is known to inhibit MAPK-activated protein kinase 2 (MK2) activity in the p38/MAPK signaling cascade, would alleviate or reduce cardiac dysfunction.</p> <p><strong>Experimental Design: </strong>Echocardiographic analysis of cardiac function of cardiac-specific Bag3 P209L transgenic (and wildtype littermate control) mice (20 – 22 months of age) was assessed by high-resolution ultrasound echocardiography (VisualSonics Vevo 2100, MS550D probe, cardiology package) to document the established disease-related cardiac dysfunction at baseline. Mice were then treated with 100mg/kg/day MMI-0100 nebulized daily for 30 days. Follow-up echocardiography was performed at 10, 20, and 30 days of MMI-0100 treatment. Echocardiographic analysis was performed to determine the systolic function (EF%, FS%), chamber dimensions, and wall thickness in systole and diastole using Vevo 2100 Workstation software package.</p> <p><strong>Results: </strong>Blinded analysis of echocardiographic data identified that Bag3 P209L Tg+ mice had a baseline cardiac dysfunction compared to wildtype controls at 20 months of age (WT= 76% EF, 44% FS; Tg+= 66% EF, 36% FS). MMI-0100 treatment significantly attenuated this dysfunction by 20 days of MMI-0100 treatment (WT= 79% EF, 47% FS; Tg+= 80% EF, 48% FS), consistent with demonstrating for the first time MK2’s role in mediating p38 signaling in the pathogenesis of Bag3 P209L cardiomyopathy.</p> <p><strong>Conclusion&nbsp;and&nbsp;Potential Impact: </strong>The MMI-0100 peptide has proven efficacious in several animal models of fibrosis driven by p38 signaling, as MK2 is a p38 downstream mediator. Future studies seek to translate the use of the MMI-0100 peptide in pediatric patients with Bag3 P209L cardiomyopathy through compassionate use FDA pathways.</p> Ashley R. Hacker, Jessica M. Berthiaume, Monte S. Willis Copyright (c) 2019 Ashley R. Hacker, Jessica M. Berthiaume, Monte S. Willis https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23478 Thu, 10 Oct 2019 10:11:51 -0400 Evaluating the Effect of Acarbose Treatment on Insulin Secretion and Sensitivity in Early Diabetes Using a Novel Interpretation of the Disposition Index Equation http://journals.iupui.edu/index.php/IMPRS/article/view/23480 <p><strong>Background and Hypothesis:</strong> <br> 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.</p> <p><strong>Background and Hypothesis:</strong> <br> 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.</p> <p><strong>Results:</strong> <br> 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.</p> <p><strong>Conclusion and Potential Impact:&nbsp;</strong></p> <p>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.</p> <p>&nbsp;</p> Clarissa Hanna, Tamara Hannon, Robert V. Considine, Kieren J. Mather Copyright (c) 2019 Clarissa Hanna, Tamara Hannon, Robert V. Considine, Kieren J. Mather https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23480 Tue, 08 Oct 2019 10:49:22 -0400 Building Public Health System Capacity for Diabetes and Hypertension Care in Western Kenya http://journals.iupui.edu/index.php/IMPRS/article/view/23482 <p><strong>Background and Hypothesis:</strong> <br> In western Kenya, estimates for prevalence of hypertension are 6-24% and diabetes are 3-5%. Complications of hypertension and diabetes are some of the fastest growing causes of morbidity and mortality in Kenya. There is limited knowledge and poor training pertaining to noncommunicable diseases (NCDs) making treatment difficult. With shortages of physicians and mid-level providers, protocol driven nursing care of hypertension and diabetes has shown to be effective. Through partnerships with counties, the NCD care program has screened 134,923 and treated 12,566 individuals for diabetes and screened 238,078 and treated 29,377 individuals for hypertension. <br> My summer project was to utilize Academic Model Providing Access to Healthcare’s (AMPATH) partnership with the Kenyan Ministry of Health (MOH) to improve access and quality of hypertension and diabetes care delivery at county MOH facilities. These improvements can be achieved through increasing our joint mentorship program led by AMPATH and county MOH staff; introducing the AMPATH Medical Record System (AMRS) into county clinics to improve care; and quality improvement with the Home Glucose Monitoring team.</p> <p><strong>Personal Role:</strong> <br> I created Standard Operating Procedures (SOPs) for Integrative Screening, Clinical Mentorship, and Home Glucose Monitoring (HGM); designed a user manual for AMRS; helped develop a M&amp;E plan for the HGM program; and assisted in the authorship of a grant to scale mentorship for diabetes and hypertension to 60 more clinical sites.</p> <p><strong>Conclusion and Potential Impact: </strong></p> <p>This project is a product of the partnership of AMPATH and the MOH to make accessible, high-quality care for hypertension and diabetes available across levels of care. By partnering with the Kenyan government and implementing care in the public sector, improvements in care are sustainable. My involvement will facilitate scaling the project to treat more patients through improving organizational capacity and helping apply for funds to implement in new areas.</p> <p>&nbsp;</p> Michael Harding, Matthew Turissini Copyright (c) 2019 Michael Harding, Matthew Turissini https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23482 Tue, 08 Oct 2019 10:49:49 -0400 Percutaneous Coronary Intervention Outcomes in Solid Organ Transplant Candidates http://journals.iupui.edu/index.php/IMPRS/article/view/23483 <p><strong>Background:</strong> 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.</p> <p><strong>Methods:</strong> 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.</p> <p><strong>Results:</strong> 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.</p> <p><strong>Conclusion:</strong> 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.</p> Kate L. Harris, BS, Kyle Frick, MD, Lawrence Lee, MD Copyright (c) 2019 Kate L. Harris, BS, Kyle Frick, MD, Lawrence Lee, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23483 Tue, 08 Oct 2019 10:50:18 -0400 What is the Optimal Time to Perform Cardiac Surgery in Patients with Stroke? http://journals.iupui.edu/index.php/IMPRS/article/view/23485 <p><strong>Background:</strong> Preoperative stroke in patients undergoing cardiac surgery is a risk factor associated with increased postoperative complications, including stroke. The optimal timing when cardiac surgery can be safely performed after stroke is unknown. The purpose of this study was to investigate how cardiac surgery timing after preoperative stroke impacts postoperative outcomes.</p> <p><strong>Methods:</strong> All patients with history of stroke who subsequently underwent cardiac surgery at IU Health Methodist Hospital between 2011-2017 were retrospectively reviewed. Patient variables were extracted from both the institutional Society of Thoracic Surgeons (STS) database and a statewide Indiana patient database. Medical records of each patient were also reviewed. Patients were grouped into cohorts by the timing of the preoperative stroke relative to cardiac surgery: Recent (within two weeks before surgery), Intermediate (greater than two weeks but less than six weeks before), and Remote (greater than six weeks before). Postoperative outcomes were reviewed and compared between groups.</p> <p><strong>Results:</strong> 165 patients were included in the study, with 48 in the Recent group, 27 in Intermediate, and 90 in Remote. 30-day mortality and postoperative stroke rates, respectively, for the groups without endocarditis (n=110) were: Recent – 0.0% and 28.6%, Intermediate – 0.0% and 8.3%, and Remote – 3.9% and 5.2%. The difference in postoperative stroke rate between Recent and Remote was significant (p=0.005) with patients in the Recent group estimated to be 7.3 times more likely to develop postoperative stroke in comparison to the Remote group, while the difference between Recent and Intermediate was not. Preoperative stroke due to endocarditis (n=55) was not associated with a significant increase in postoperative stroke, regardless of timing of surgery.</p> <p><strong>Conclusion:</strong> Delaying cardiac surgery for six weeks after stroke occurrence may lead to significantly reduced likelihood of postoperative stroke. The exception would be patients with preoperative stroke due to septic emboli from endocarditis, as timing of cardiac surgery does not seem to affect postoperative mortality or stroke rates in these patients.</p> Timothy Hartman, BS, Mackenzie Madison, MS, Niha Namburi, MHA, Luke Haag, Lawrence Lee, MD Copyright (c) 2019 Timothy Hartman, BS, Mackenzie Madison, MS, Niha Namburi, MHA, Luke Haag, Lawrence Lee, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23485 Thu, 10 Oct 2019 10:18:23 -0400 Impact of a Long-Term High Fat Diet on Bone Microarchitecture and Muscle Structure in Adult Male and Female Normal Mice http://journals.iupui.edu/index.php/IMPRS/article/view/23486 <p><strong>Background and Hypothesis:</strong> <br> Hyperglycemia is a major source of disease and morbidity among the adult population. Prior studies correlate long-term high fat diet (HFD) mediated hyperglycemia with bone fragility and muscle weakness. Furthermore, the mechanism driving hyperglycemia between sexes are unknown. Our group previously showed that HFDs induced insulin resistance in male mice and glucose intolerance in female mice. This establishes the need to study the impact of long-term HFDs on the bones and muscles using an older cohort of both male and female mice. For that, we hypothesized a long-term HFD mediated hyperglycemia will change bone and muscle structures and impair their functions in adult male and female mice.</p> <p><strong>Experimental Design or Project Methods:</strong> <br> 22-week C57Bl6 mice were fed either a HFD or low fat diet (LFD) for 25 weeks. After euthanasia, bones and muscles were harvested and evaluated using MicroCT, histology, and mechanical testing. Statistical analysis was performed using GraphPad Prism with p&lt;0.05 considered significant.</p> <p><strong>Results:</strong> <br> MicoCT data saw significant reductions to cortical thickness (p&lt;0.05), bone mineral density (p&lt;0.001), and increases to medullary area (p&lt;0.05) among HFD males and females compared to LFD. HFD-males also experienced significant increase in cortical porosity (P&lt;0.001) whereas no changes were noted in HFDfemales. Trabecular bone volume was relatively unchanged. HFD increased cortical osteoclast surface (p&lt;0.001) for both sexes. Bone histology saw increased marrow adiposity among HFD-females (p&lt;0.05). Muscle histology exhibited HFD-related reductions in myofiber diameter (p&lt;0.001) for both sexes. Mechanical testing demonstrated reduced young’s modulus (p&lt;0.05) and yield stress (p&lt;0.05) among HFD mice, despite non-significant differences in ultimate strength.</p> <p><strong>Conclusion and Potential Impact:</strong> The changes associated with a long-term HFD differed between sexes but still led to functional impairments of bone and muscle for both sexes, emphasizing the importance of looking further into the mechanisms responsible for these changes. This can potentially translate to the clinic in the treatment of musculoskeletal complications associated with HFDs.</p> Weston He, Trupti Trivedi, Gabriel Pagnotti, Sreemala Murthy, Yun She, Sutha John, Jack Truitt, Khalid S. Mohammad, Theresa A. Guise Copyright (c) 2019 Weston He, Trupti Trivedi, Gabriel Pagnotti, Sreemala Murthy, Yun She, Sutha John, Jack Truitt, Khalid S. Mohammad, Theresa A. Guise https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23486 Tue, 08 Oct 2019 10:51:10 -0400 Long Term Outcomes of Iliac Artery Aneurysm (IAA) and Abdominal Aortic Aneurysm (AAA) Repair with Endovascular Procedures at the Parkview Heart Institute http://journals.iupui.edu/index.php/IMPRS/article/view/23487 <p><strong>Background:</strong> Favorable characteristics of the minimally invasive endovascular aneurysm repair (EVAR), such as a reduced recovery time, have led physicians and patients alike to elect for this procedure over open repair. Longitudinal study on endovascular repair has raised concerns regarding an increased rate of late failure leading to rupture and overall higher rates of re-intervention for patients.</p> <p><strong>Objective:</strong> This project compares the outcomes of endovascular stent graft repairs of AAA and IAA at Parkview Heart Institute over a certain time span.</p> <p><strong>Methods &amp; Design:</strong> This study is a retrospective review of patients (N=89) with AAA/IAA repair since 2014. The EPIC online medical record is the main tool in data extraction while statistical analysis was conducted via Microsoft Excel (p = 0.05).</p> <p><strong>Results:</strong> Males, ages 51-75 y.o., White population, non-Hispanics, and BMI rating of 1-30 were observed to have more AAAs. A significant decrease in length of stay over historical averages was observed with at most 1-3 days and minimal complications. Most causes for re-interventions were due to endo-leaks. More than 60% of patients with AAA and EVAR have a significant past medical history of smoking, hypertension, and chronic obstructive pulmonary disease. Finally, most pre- and post-operative aneurysms sizes are within 41-60 mm in diameter.</p> <p><strong>Conclusion and Potential Impact:</strong> This study analyzed 89 patients that experienced AAA instead of IAA. Certain populations, such as being male, ages 51-75 y.o., White population, non-Hispanics, and BMI rating of 1-30 with past medical history characteristics are frequent in patients with EVAR. The procedure has minimal complications in comparison to open abdominal surgery, and this study will add to the literature by aiding physicians’ determination of the best course of action for treating patients with AAA/IAA.</p> Douglas Gray, M.D., Christopher Herrera, B.S., Kim Recht, N.P. Copyright (c) 2019 Douglas Gray, M.D., Christopher Herrera, B.S., Kim Recht, N.P. https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23487 Tue, 08 Oct 2019 11:05:55 -0400 Characterization of RNA binding proteins required for receptor-mediated endocytosis in C. elegans http://journals.iupui.edu/index.php/IMPRS/article/view/23489 <p><strong>Background and Hypothesis:</strong> Recently, a study conducted by the Hundley lab revealed that mutation of a specific RNA editing protein, <em>ADR2</em>, in the model organism <em>C. elegans</em> increased receptor-mediated endocytosis of yolk protein <em>vit-2</em> and oocyte maturation. Of interest to us was what additional RNA binding proteins were involved in the expression of <em>vit-2.</em> 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 <em>vit-2.</em></p> <p><strong>Project Methods:</strong> Previously, <em>adr-2(-) C. elegans</em> strain expressing a yolk protein (<em>vit-2</em>) fused to GFP and a library of <em>E. coli</em> strains expressing RNAi directed to specific RNA binding proteins were engineered. <em>vit-2:GFP</em> 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 <em>vit-2</em> endocytosis.</p> <p><strong>Results:</strong> 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 <em>gld-1</em> and <em>puf-8</em> 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 <em>vit-2</em> expression.</p> <p><strong>Conclusion and Potential Impact:</strong> This screen successfully gave direction to future studies of <em>ADR-2</em> 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 <em>vit-2</em> 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 <em>adr-2(-)</em> worms. This would add to what is known about ADARs and life span and possibly demonstrate a role for ADARs in germ cell maturation.</p> <p>&nbsp;</p> Margaret D. Holdeman, Farida Daouda, Heather A. Hundley, Ph.D. Copyright (c) 2019 Margaret D. Holdeman, Farida Daouda, Heather A. Hundley, Ph.D. https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23489 Tue, 08 Oct 2019 11:10:15 -0400 Targeting the Warburg effect to overcome lapatinib resistance in esophageal adenocarcinoma http://journals.iupui.edu/index.php/IMPRS/article/view/23494 <p><strong>Background:</strong> Esophageal adenocarcinoma (EAC) overexpresses HER2. Lapatinib, a dual tyrosine kinase inhibitor blocking HER1 and HER2 pathways failed to improve patient survival. Molecular mechanisms of this lapatinib resistance are still unclear. We therefore explored the role of glycolytic enzyme pyruvate kinase M2 (PKM2), a key regulator of the Warburg effect in the lapatinib resistance mechanism of EAC cells.</p> <p><strong>Methods:</strong> First we established a lapatinibresistant OE19 (LPR-OE19) cell line from OE19 EAC cells. We then investigated the comparative cell growth inhibition, apoptotic and lactate production effects of HER2 inhibitor lapatinib and PKM2 inhibitor shikonin either alone or in combinations, both in OE19 and LPR-OE19 cells with and without knockdown of PKM2 and HSP40 by siRNAs.</p> <p><strong>Results:</strong> Lapatinib resistant LPR-OE19 cells showed downregulation of HSP40 both at protein and mRNA levels, but showed upregulation of PKM2 only at the protein level. LPR-OE19 cells showed significantly reduced sensitivity to lapatinib induced cell growth inhibition, apoptosis and decreased cell migration compared to parent OE19 cells. Interestingly, augmented cell growth inhibition, apoptosis and decreased cell migration were observed in LPR-OE19 cells compared to parent OE19 cells when lapatinib was combined with shikonin. Knockdown of PKM2 in LPR-OE19 cells abolished the reduced sensitivity of lapatinib induced cell growth inhibition, and also abolished augmented cell growth inhibition response when lapatinib and shikonin were combined. Contrary to that, knockdown of HSP40 in OE19 cells enhanced PKM2 expression leading to significantly reduced sensitivity to lapatinib induced growth inhibition with augmented growth inhibition when lapatinib and shikonin were combined. Moreover, LPR-OE19 cells showed enhanced lactate production compared to parent OE19. Interestingly, PKM2 and HSP40 co-localize with each other in the cell nucleus suggesting that PKM2 binds to molecular chaperone HSP40.</p> <p><strong> Conclusions:</strong> These data suggest that targeting HSP40PKM2 interaction could be an innovative therapeutic approach to overcome lapatinib resistance in EAC.</p> Samantha Holmes, Sazzad Hassan, MD, Victoria Makuru, Urs von Holzen von Holzen Copyright (c) 2019 Samantha Holmes, Sazzad Hassan, MD, Victoria Makuru, Urs von Holzen von Holzen https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23494 Tue, 08 Oct 2019 11:10:45 -0400 Efficiency and Blood Loss of Various Spacer and Intramedullary Dowel Constructs in Two-Stage Treatment of Total Knee Arthroplasty http://journals.iupui.edu/index.php/IMPRS/article/view/23495 <p><strong>Background and Hypothesis:</strong> Treatment for infected total knee arthroplasty (TKA) employs antibiotic-eluding articulating or static spacers, with or without intramedullary (IM) dowels between implant resection and reimplantation. While it is unknown which spacer type is more efficient intra-operatively, IM dowels require additional time for fabrication. Surgical efficiency is critical to minimizing anesthesia time and blood loss, especially in complex surgeries with compromised hosts. We quantified operative time and postoperative intra-articular blood loss based on spacer type and the use of IM dowels.</p> <p><strong>Project Methods:</strong> 103 consecutive infected TKAs treated from 2010-2019 were retrospectively reviewed. Outcome variables included operative time and intraarticular drain rate. Covariates included sex; age, BMI; ASA-PS classification; surgeon; McPherson infection classification; tourniquet time; tranexamic acid (TXA) use; intrathecal anesthesia, length of stay, and blood transfusion. Multivariate analyses were used.</p> <p><strong>Results:</strong> The sample was 52% female with average age of 66±9 years and average BMI of 36±9 kg/m<sup>2</sup>. Articulating spacers without dowels (ASwoD), articulating spacers with dowels (ASwD), and static spacers with dowels were used in 57.3%, 21.4%, and 21.4% of knees, respectively. Longer mean operating time was observed when static spacers with dowels were used at resection (162 vs.130 ASwoD/140 ASwD minutes;<em> p</em>=0.001) and reimplantation (187 vs. 149 ASwoD/148 ASwD minutes; p=0.017). At reimplantation, drain rate was highest when articulating spacers with dowels were used (37 vs. 20/26 mL/hr), but not when TXA was used (<em>p</em>=0.002).</p> <p><strong>Conclusion and Potential Impact:</strong> Articulating and static spacers provide equivalent infection eradication, and the necessity of IM dowels has not been thoroughly studied. In light of this equivalency, it is important to understand other costs associated with spacer types and IM dowels. Our observations that spacer/dowel constructs affect time under anesthesia and blood loss may contribute to the efficiency and safety of the two-stage treatment protocol.</p> Hanna House, BS, Mary Ziemba-Davis, BA, Michael Meneghini, MD Copyright (c) 2019 Hanna House, BS, Mary Ziemba-Davis, BA, Michael Meneghini, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23495 Tue, 08 Oct 2019 11:11:12 -0400 Neonatal bowel resections occurring in the NICU are associated with lower survival rates compared to the OR http://journals.iupui.edu/index.php/IMPRS/article/view/23496 <p><strong>Background and Hypothesis:</strong> Neonatal surgical procedures frequently occur at bedside in the neonatal intensive care unit (NICU) rather than in the operating room (OR). The goal of this study was to isolate risk factors for neonatal bowel resections occurring in the NICU, compare outcomes of bowel resections occurring both in the NICU and OR, and evaluate modifiable factors in both locations. </p> <p><strong>Experimental Design or Project Methods:</strong> We reviewed 139 neonatal (&lt;30 days old) patients at Riley Hospital for Children who underwent bowel resection in either the NICU or the OR from 2014-2018. Statistical analysis included bivariate, multivariable, and propensity score matching analyses. </p> <p><strong>Results:</strong> When compared to neonates who had bowel resection in the OR, the NICU group had lower mean gestational age (EGA) at birth (26 vs 34 weeks, p=&lt;0.0001), lower weight at surgery (72.7% NICU patients &lt;1000 grams vs 7.6% OR patients, p=&lt;0.0001), and had the operative diagnosis of spontaneous intestinal perforation (48.8%) or NEC (42.9%) compared to OR patients (18.7%) for both diagnoses (p=&lt;0.0001, p=0.0003). Bivariate analysis revealed that EGA at birth (OR 0.668, p=&lt;0.0001), APGAR 1 minute (OR 0.776, p=0.001), and pre-op respiratory FiO2 percentage (OR 2.514, p=&lt;0.0001) were significant predictors of operation location. Propensity score matching (15 matched pairs) showed survival rates were lower in the NICU group (p=0.0156). Modifiable risk factors, such as the usage of a heating device during surgery (p=&lt;0.0001) and temperature monitoring during surgery (p=&lt;0.0001) were positively correlated with survival, while blood administration in the first 24 hours post-surgery (p=0.001) was correlated with worse survival.</p> <p><br> <strong>Conclusions:</strong> There are definable characteristics of neonates who have bowel resections in the OR vs NICU, but the primary outcome difference between these groups is survival. We also identified modifiable factors that can be studied to further improve survival of patients with operations in both settings.</p> <p>&nbsp;</p> Lauren Howser, Carly Goehring, Cassandra Anderson, Eamaan Turk, Yan Han, George Eckert, Dr. Brian Gray, MD Copyright (c) 2019 Lauren Howser, Carly Goehring, Cassandra Anderson, Eamaan Turk, Yan Han, George Eckert, Dr. Brian Gray, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23496 Tue, 08 Oct 2019 11:18:37 -0400 Pediatric Anticholinergic Toxidrome and Treatment with Physostigmine http://journals.iupui.edu/index.php/IMPRS/article/view/23497 <p><strong>Background and Hypothesis:</strong> 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.</p> <p><strong>Project Methods:</strong> 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.</p> <p><strong>Results:</strong> 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.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> Sarah Huber, Bob Avera, MD, Adam Overberg PharmD, BCPS, CSPI, Shannon Morton, MPH, Kristine Nanagas MD Copyright (c) 2019 Sarah Huber, Bob Avera, MD, Adam Overberg PharmD, BCPS, CSPI, Shannon Morton, MPH, Kristine Nanagas MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23497 Tue, 08 Oct 2019 11:19:01 -0400 Surface-based analysis of cortical thickness and volume loss in Alzheimer’s disease http://journals.iupui.edu/index.php/IMPRS/article/view/23524 <p><strong>Background and Hypothesis:</strong> Magnetic resonance imaging (MRI) has become a useful tool in monitoring the progression of Alzheimer's disease. Previous surface-based analysis has focused on changes in cortical thickness associated with the disease<sup>1</sup>. The objective of this study is to analyze MRI-derived cortical reconstructions for patterns of atrophy in terms of both cortical thickness and cortical volume. We hypothesize that Alzheimer’s Disease progression will be associated with a more significant change in volume than thickness.</p> <p><strong>Experimental Design or Project Methods:</strong> MRI data was obtained from the Alzheimer's Disease Neuroimaging Initiative (ADNI). All subjects with baseline and two-year 3T MRI scans were included. Segmentation of MRIs into gray and white matter was performed with FreeSurfer<sup>2,3,4,5</sup>. Subjects whose scans did not segment accurately were excluded. Surfaces were then registered to a common atlas with Ciftify<sup>6</sup>, and anatomically-constrained Multimodal Surface Matching (aMSM) was used to analyze longitudinal changes in each subject<sup>7</sup>. This produced continuous surface maps showing changes in cortical surface area and thickness. These maps were multiplied to create cortical volume maps<sup>8</sup>. Permutation Analysis of Linear Models (PALM) was used to perform two-sample t-tests comparing the maps of the Alzheimer’s and control groups<sup>9</sup>.</p> <p><strong>Results:</strong> Preliminary analysis of nine Alzheimer’s subjects and nine control subjects produced surface maps displaying patterns that were expected given previous research findings<sup>10,11</sup>. There was increased volume and thickness loss in Alzheimer’s subjects relative to controls, with relatively high loss in structures of the medial temporal lobe. Future analysis of a larger sample will determine whether statistically significant differences exist between the Alzheimer’s and control groups in terms of thickness loss and volume loss.</p> <p><strong>Conclusion and Potential Impact:</strong> If significant results are found, surface-based analysis of cortical volume may allow for detection of atrophy at an earlier stage in disease progression than would be possible based on cortical thickness.</p> <p>&nbsp;</p> <p><strong>References</strong></p> <p>1. Clarkson MJ, Cardoso MJ, Ridgway GR, Modat M, Leung KK, Rohrer JD, Fox NC, Ourselin S. A comparison of voxel and surface based cortical thickness estimation methods. NeuroImage. 2011 Aug 1; 57(3):856-65.</p> <p>2. Dale AM, Fischl B, Sereno MI. Cortical surface-based analysis. I. Segmentation and surface reconstruction. Neuroimage. 1999;9:179194.</p> <p>3. Fischl B, Sereno M, Dale A. Cortical surface-based analysis. II: Inflation, flattening, and a surface-based coordinate system. Neuroimage. 1999;9:195–207.&nbsp;</p> <p>4. Fischl B, Salat DH, Busa E, Albert M, Dieterich M, Haselgrove C, van der Kouwe A, Killiany R, Kennedy D, Klaveness S, Montillo A, Makris N, Rosen B, Dale AM. Whole brain segmentation: automated labeling of neuroanatomical structures in the human brain. Neuron 2002;33:341-355.</p> <p>5. Fischl B, Salat DH, van der Kouwe AJ, Makris N, Segonne F, Quinn BT, Dale AM. Sequence-independent segmentation of magnetic resonance images. Neuroimage 2004;23 Suppl 1:S69-84.</p> <p>6. Glasser MF, Sotiropoulos SN, Wilson JA, Coalson TS, Fischl B, Andersson JL, Xu J, Jbabdi S, Webster M, Polimeni JR, Van Essen DC, Jenkinson M, WU-Minn HCP Consortium. The minimal preprocessing pipelines for the Human Connectome Project. Neuroimage. 2013 Oct 15;80:105-24.</p> <p>7. Robinson EC, Garcia K, Glasser MF, Chen Z, Coalson TS, Makropoulos A, Bozek J, Wright R, Schuh A, Webster M, Hutter J, Price A, Cordero Grande L, Hughes E, Tusor N, Bayly PV, Van Essen DC, Smith SM, Edwards AD, Hajnal J, Jenkinson M, Glocker B, Rueckert D. Multimodal surface matching with higher-order smoothness constraints. Neuroimage. 2018;167:453-65.</p> <p>8. Marcus DS, Harwell J, Olsen T, Hodge M, Glasser MF, Prior F, Jenkinson M, Laumann T, Curtiss SW, Van Essen DC. Informatics and data mining tools and strategies for the human connectome project. Front Neuroinform 2011;5:4.</p> <p>9. Winkler AM, Ridgway GR, Webster MA, Smith SM, Nichols TE. Permutation inference for the general linear model. NeuroImage, 2014;92:381-397</p> <p>10. Matsuda, H. MRI morphometry in Alzheimer’s disease. Ageing Research Reviews. 2016 Sep;30:17-24.</p> <p><strong></strong>11. Risacher SL, Shen L, West JD, Kim S, McDonald BC, Beckett LA, Harvey DJ, Jack CR Jr, Weiner MW, Saykin AJ. Alzheimer's Disease Neuroimaging Initiative (ADNI). Longitudinal MRI atrophy biomarkers: relationship to conversion in the ADNI cohort. Neurobiol Aging. 2010 Aug;31(8):1401-18.&nbsp;</p> Emily Iannopollo, Ryan Plunkett, Kara Garcia Copyright (c) 2019 Emily Iannopollo, Ryan Plunkett, Kara Garcia https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23524 Tue, 08 Oct 2019 11:19:37 -0400 Parents’ Descriptions of Neonatal Palliation as a Treatment Option Prior to Periviable Delivery http://journals.iupui.edu/index.php/IMPRS/article/view/23498 <p><strong>Background and Hypothesis:</strong> During periviable deliveries, parents are confronted with overwhelming and challenging decisions, about which they may know little. This study aimed to explore qualitatively the language that mothers and important others (IOs) utilize when discussing palliation, or ‘comfort care,’ as a treatment option in the context of periviability.</p> <p><strong>Project Methods:</strong> We prospectively recruited pregnant women (and designated IOs) admitted to labor and delivery for a threatened periviable delivery (22-25 weeks GA) at two hospitals between September 2016 and January 2018. Using a semi-structured interview guide, we explored participants’ perceptions of palliation and neonatal treatment options. Women were asked items such as, “How was the choice of resuscitation presented to you?” and “What were the options presented?” Research assistants developed a codebook for the interview transcripts, and NVivo 12 was used for qualitative analysis. </p> <p><strong>Results:</strong> Thirty women and 16 IOs were recruited in total. Participants’ descriptions of palliation fell into five broad categories – ‘doing nothing,’ ‘not resuscitating,’ ‘withdrawal of care,’ ‘implicit comfort care,’ and ‘explicit comfort care.’ The majority of parents perceived comfort care not as a distinct treatment option, but rather as the absence of treatment. Several barriers to the comprehension of comfort care were observed, including subjects’ unfamiliarity with the terminology or the inability to remember its explanation during antenatal consultation. Parents described comfort care with either neutral or negative connotations, and even those parents able to discuss the concept neutrally chose resuscitation as their preferred treatment option.</p> <p><strong>Conclusion and Potential Impact:</strong> This study revealed that many parents facing periviable delivery may lack understanding of comfort care as a neonatal treatment option. These parental perceptions highlight the need to improve counseling efforts in order to maximize parents’ informed decision-making.</p> Shannon Jager, Brownsyne Tucker Edmonds, MD, MPH, MS, Shelley Hoffman, MPH, Erin Jeffries, MD, MS, Tatiana Laitano, MD, Karen Kavanaugh, PhD, RN, FAAN Copyright (c) 2019 Shannon Jager, Brownsyne Tucker Edmonds, MD, MPH, MS, Shelley Hoffman, MPH, Erin Jeffries, MD, MS, Tatiana Laitano, MD, Karen Kavanaugh, PhD, RN, FAAN https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23498 Tue, 08 Oct 2019 11:20:09 -0400 A Multi-Disciplinary Care Model for People with Drug-Resistant HIV: The AMPATH HIV Drug Resistance Clinic in Kenya http://journals.iupui.edu/index.php/IMPRS/article/view/23499 <p><strong>Background:</strong> 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 “2<sup>nd</sup> 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.</p> <p><strong>Methods:</strong> 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.</p> <p><strong>Personal role:</strong> 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.</p> <p><strong>Moving Forward: </strong>By building the capacity of clinical leaders in decentralized regional AMPATH sites, we hope to expand the DRC care model within the AMPATH network.&nbsp;</p> Bilal Jawed, John Humphery, Adrian Gardner Copyright (c) 2019 Bilal Jawed, John Humphery, Adrian Gardner https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23499 Tue, 08 Oct 2019 00:00:00 -0400 Physician Receptiveness of Ventilation-Perfusion Imaging in a Randomized Clinical Trial http://journals.iupui.edu/index.php/IMPRS/article/view/23500 <p><strong>Background and Hypothesis:</strong> Computed Tomography of the pulmonary arteries (CTPA) is the most common imaging modality for evaluating patients for suspected pulmonary embolism (PE), but carries the risk of acute kidney injury (AKI) from contrast media exposure. In appropriately selected patients, ventilation scintigraphy (VQ) imaging is a diagnostically equivalent alternative. We hypothesized that physician perceptions of diagnostic accuracy and study availability contribute to under-utilization of VQ imaging.</p> <p><strong>Project Methods:</strong> Patients with suspected PE at increased risk of acute kidney injury, were randomly selected to undergo VQ instead of CTPA. Patients unable to consent, patients with a history of pulmonary surgery, and those undergoing contrastenhanced imaging for other indications were excluded. A screening chest radiograph was obtained prior to study imaging allocation. All cases were reviewed by a nuclear medicine radiologist blinded to acceptance or refusal of VQ imaging allocation. The primary outcome was defined as the rate of physician-refusal of VQ imaging. The unprompted physician-reported reason for refusal was recorded, in real-time, along with any other general responses.</p> <p><strong>Results:</strong> Following exclusions, 42 subjects were enrolled. Notably, chest radiograph findings excluded only 2 subjects. The reviewing nuclear radiologist agreed with all study-selections for VQ appropriateness and there was no instance of nondiagnostic VQ imaging. Treating physicians refused VQ imaging randomization in 48% (20/42). Physicians also believed VQ imaging lacked sufficient diagnostic accuracy in the context of active non-pulmonary malignancy in 29% (12/42) of cases. Although CT did not identify cases not seen on chest radiograph, in 12% (5/42) cases suspected pneumonia was the reason for refusal. Statements such as “VQ is inferior [for PE],” and “VQ takes too long” were characteristic of general responses from treating providers.</p> <p><strong>Conclusion and Potential Impact:</strong> VQ imaging remains under-utilized in patients at risk of AKI. Perceived limitations to diagnostic accuracy and study availability are contributors to under-utilization.</p> Colton Junod, BS, Alice M. Mitchell, MD, MS Copyright (c) 2019 Colton Junod, BS, Alice M. Mitchell, MD, MS https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23500 Tue, 08 Oct 2019 14:14:54 -0400 Patient "GRIT" Does Not Predict Pain and Opioid Consumption Following Total Knee Arthroplasty http://journals.iupui.edu/index.php/IMPRS/article/view/23503 <p><strong>Background and Hypothesis:</strong> TKA results in severe postoperative pain and differential needs for narcotics. Medical covariates of pain and opioid use following TKA have been investigated, but the influence of personality has not. GRIT consists of passion/consistency of interest in goals and perseverance of effort. Perseverance has been associated with beneficial autonomic nervous system activity. GRIT has been associated with independence in activities of daily living, decreased depression/anxiety, and increased well-being in chronically ill patients. We evaluated whether GRIT modulates pain and opioid consumption following TKA, hypothesizing that higher GRIT scores would be associated with lower pain and opioid consumption.</p> <p><strong>Project Methods:</strong> The GRIT survey was retrospectively administered to 254 consecutive patients with cemented TKAs performed between 2016 and 2018 by a single surgeon using the same pain protocols and implant. Results were paired with prospectively collected average pain score during the first 24 hours following PACU discharge, average pain during the remainder of hospital stay, final pain score prior to discharge, time to first opioid medication after PACU discharge, total opioid consumption in morphine equivalents (MEQs) during the first 24 hours after PACU, and average MEQs/day during the remainder of stay.</p> <p><strong>Results:</strong> The sample was 71% female with average age of 67.5±8.3 years and average BMI of 33.7±7.2 kg/m2. Coefficient r for pain metrics and passion, perseverance, and GRIT ranged from -0.145 to 0.078 (<em>p</em>≥0.146) with the exception of a small (0.136), but significant (<em>p</em>=0.032) correlation between final pain score and perseverance. r for opioid metrics and passion, perseverance, and GRIT ranged from -0.039 to 0.087 (<em>p</em>≥0.172).</p> <p><strong>Conclusion and Potential Impact:</strong> Mean GRIT scores ranged from 3.7-4.3 with a median range of 3.8-4.3, suggesting demand characteristics associated with the instrument. Post hoc statistical power was low (≈35%) given n and small effect sizes. It is hoped, however, that our study fosters investigation of individual characteristics and important clinical outcomes.</p> Sydney Keller, Sachin Seetharam, Mary Ziemba-Davis, R. Michael Meneghini, MD Copyright (c) 2019 Sydney Keller, Sachin Seetharam, Mary Ziemba-Davis, R. Michael Meneghini, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23503 Tue, 08 Oct 2019 14:15:26 -0400 Vision Dysfunction in Circadian Clock Gene Bmal Mice http://journals.iupui.edu/index.php/IMPRS/article/view/23506 <p><strong>Background and Hypothesis:</strong> 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 (<em>Bmal1</em>), 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.</p> <p><strong>Experimental Design or Project Methods:</strong> The Bmal<sup>+/-</sup> mice were inbred and genotyped to obtain wild-types (WT), Bmal<sup>+/-</sup>, and Bmal<sup>-/-</sup>. 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.</p> <p><strong>Results:</strong> 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<sup>+/-</sup>, and Bmal-/- when compared to the WT group. Under photopic conditions, the circadian peak of ERG amplitude was reversed for Bmal<sup>-/-</sup> when compared to both WT and Bmal<sup>+/-</sup> mice. The OKR assessment was decreased substantially for Bmal<sup>+/-</sup> (0.3748c/d), and Bmal<sup>-/-</sup> (0.3130c/d) as compared to the WT mice (0.4827c/d).&nbsp;</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> Humza Khan, Deepa Mathew, Qianyi Luo, Ashay Bhatwadekar Copyright (c) 2019 Humza Khan, Deepa Mathew, Qianyi Luo, Ashay Bhatwadekar https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23506 Tue, 08 Oct 2019 14:15:57 -0400 Hemoglobin Expression and Function in Human Corneal Epithelium http://journals.iupui.edu/index.php/IMPRS/article/view/23535 <p>The corneal epithelium forms the outer layer of the cornea. It provides mechanical protection, prevents fluid loss and forms a barrier to invasive pathogens. Though hemoglobin expression has been extensively studied in erythroid cells, recent work in multiple cell systems has documented hemoglobin expression in cells of non-erythroid origin. However, the function of hemoglobin in non-erythroid cells has yet to be established. The hypothesis that hemoglobin is expressed in corneal epithelium and that it functions to protect against oxidative stress will be examined in the present work. Hemoglobin expression and function was examined by immunocytochemistry and western blot analysis. Native human corneal explants and an immortalized human corneal epithelial cell line were examined. Expression of hemoglobin beta and delta chains was demonstrated at the protein level in both native and cell culture preparations. Hexamethylene bisacetamide is a known inducer of hemoglobin beta chain expression in murine erythroleukemia cells. HMBA did function to increase beta chain expression in cultured corneal epithelium as well. Beta chain immunolocalization was primarily cytoplasmic, while the delta chain localized to both cytoplasmic and membrane domains. Oxidative stress, from hydrogen peroxide exposure, was shown to upregulate delta chain expression. In conclusion, hemoglobin chains are expressed in corneal epithelium and could function to protect against oxidative stress. This is relevant given that exposure to light and high oxygen tensions render the cornea particularly susceptible to oxidative damage.</p> Ryan McBride, Erin Perez, Ernest Talarico, Brian Kennedy Copyright (c) 2019 Ryan McBride, Erin Perez, Ernest Talarico, Brian Kennedy https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23535 Wed, 09 Oct 2019 09:27:51 -0400 Impact of T Helper Cell Subtype on CAR-T Cell Therapy Efficacy http://journals.iupui.edu/index.php/IMPRS/article/view/23509 <p><strong>Background and Hypothesis:</strong> Chimeric antigen receptors (CARs) are recombinant receptors with high affinity for the target antigen. Used for tumor therapy, CARs are transduced into patient T cells. CAR-T cells specific for CD19 are used to treat B cell acute lymphoblastic leukemia (B-ALL). Cancerous B cells are destroyed by CAR-T cells in an antigen-specific manner. Currently being used in conjunction with radiation and other cancer therapies to prohibit relapse, Dr. Marco Davila of the Moffitt Cancer Center, has shown that CAR-T therapy induces long term remission and B cell aplasia.</p> <p><strong>Experimental Design:</strong> In this experiment the CAR vector obtained from Dr. Davila was transduced into T helper cells cultured under varying conditions (Th0, Th9, and ThGranzyme A). B cell killing and longevity of transduced CAR-Th cells were monitored as part of the criteria for determining the most effective Th subtype for the CAR-T therapy. The target cell killing-mechanism was analyzed at the RNA level using quantitative polymerase chain reaction (qPCR) to analyze gene expression of cytotoxic molecules including granzymes A/B, perforins, Fas-FasL, and TNF-α. Th9 cells were expected to be among the most effective of the indicated subtypes due to their longevity and coordination of the immune response.</p> <p><strong>Results:</strong> T cells in all conditions were effectively transduced for CAR expression, although Th9 cells demonstrated a greater proportion of cultured cells that were transduced with the CAR. QPCR results suggest that there is specification of cytotoxic programs among the culture conditions. In Th9 cells, qPCR results suggest their use of perforin and TNF-α. Ongoing studies will compare cytotoxic activity.</p> <p><strong>Potential Impact:</strong> Further steps after determining the most effective culture conditions include injecting transduced Th cells of the optimized subtype into mice afflicted with BALL to assess cancer killing in vivo as well as the potential harm of the therapy to the patient.</p> Ibrahim M. Khan, BS BA, Andrew S. Nelson, PhD, Mark H. Kaplan, PhD Copyright (c) 2019 Ibrahim M. Khan, BS BA, Andrew S. Nelson, PhD, Mark H. Kaplan, PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23509 Tue, 08 Oct 2019 14:16:33 -0400 Impact of IVC Filter Guidelines, Registry, and Clinic on Filter Retrieval. http://journals.iupui.edu/index.php/IMPRS/article/view/23511 <p><strong>Background and hypothesis:</strong> Deep vein thrombosis (DVT) and pulmonary embolism (PE), collectively referred to as venous thromboembolism (VTE), are serious medical conditions that affect up to 900,000 Americans yearly, accounting for up to 100,000 deaths. The first line treatment for VTE is anticoagulation; however, in patients who experience a contraindication to, or failure of anticoagulation, an IVCF may be used. There are two types of IVCFs, permanent and retrievable. Retrievable filters are indicated when the contraindication to anticoagulation is transient, and they may be removed once the contraindication has passed. Retrievable filters have become associated with serious complications such as filter fracture, migration, and IVC perforation. Subsequently, they have become the subject of litigation. As such, strategies should be undertaken to reduce filter dwell time and improve filter retrieval rates. We hypothesize that implementation of IVCF guidelines, registry, and clinic will reduce dwell time while increasing retrieval rate.</p> <p><strong>Methods:</strong> This study was a mixed retrospective and prospective chart review of patients who received an IVCF before and after implementation of IVCF guidelines, registry, and clinic. The guidelines, registry, and clinic were established in July 2017. Cases were analyzed during the years 2014-2015 (n=191) and 2017-2018 (n=103) beginning in July 2017. Data was obtained on filter retrieval rate, dwell time, filter-associated complications, and indication for placement.</p> <p><strong>Results:</strong> There was a significant decrease in dwell time (p&lt;.001) and a significant increase in retrieval rate (p&lt;.001). There was no difference in complication rate, and there was a decrease in filter placement in patients with ‘soft’ indications, though this difference was not statistically significant (p=.109).</p> <p><strong>Conclusion and potential impact:</strong> Implementation of dedicated efforts to increase patient follow-up and filter retrieval were effective in reducing dwell time and retrieval rate. Although there was no significant difference in complication rate, these efforts may be protective against litigation for patients who experience a filter-associated complication.</p> Brandon Kiley, BS, Lisa Hollister, MSN, RN, T. Eric White, MD, Emily Keltner, BS, MA, Thein Zhu, MBBS, FACE, FRCP, Dazar Opoku, MPH Copyright (c) 2019 Brandon Kiley, BS, Lisa Hollister, MSN, RN, T. Eric White, MD, Emily Keltner, BS, MA, Thein Zhu, MBBS, FACE, FRCP, Dazar Opoku, MPH https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23511 Tue, 08 Oct 2019 14:17:00 -0400 Intraperitoneal Mesalamine Does Not Restores Mesenteric Perfusion or Prevent Mucosal Injury Following Intestinal Ischemia and Reperfusion http://journals.iupui.edu/index.php/IMPRS/article/view/23512 <p><strong>Background and Hypothesis:</strong> 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 <strong>that administration of mesalamine prior to intestinal ischemia would reduce epithelial cell damage by I/R and restore mesenteric perfusion.</strong></p> <p><strong>Project Methods:</strong> 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&amp;E and scored for intestinal mucosal injury. Data were assessed for normalcy and compared by Mann-Whitney-U test. P&lt;.05 was significant.</p> <p><strong>Results:</strong> 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.</p> <p><strong>Conclusion:</strong> 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.</p> Thomas Knowles, Brian Hosfield, Chris Shelley, Troy Markel Copyright (c) 2019 Thomas Knowles, Brian Hosfield, Chris Shelley, Troy Markel https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23512 Tue, 08 Oct 2019 14:17:29 -0400 Calcifications due to Mitral Valve Disease Induced by Rheumatoid Arthritis in K/B.g7 Mice http://journals.iupui.edu/index.php/IMPRS/article/view/23513 <p><strong>Background and Hypothesis:</strong> Rheumatoid arthritis (RA) is a rheumatic autoimmune disease wherein the host generates self-antibodies that target the synovial lining of joints. However, it’s been observed that patients with RA also develop Mitral Valve Disease (MVD), although the mechanism is not very well known. In my project, I focused on the physiological changes of the heart during the formation of MVD, specifically focusing on calcifications and whether they form on the inflamed valve. I hypothesized that calcifications are developing during disease progression, resulting in advancement of the inflammation cycle.</p> <p><strong>Experimental Design or Project Methods:</strong> A total of thirteen mouse hearts were removed and stored in OTC at -80 C: 5 K/B.g7 (7 months), 5 K/B.g7 (8 weeks) and 3 B6 (1.5 years). The hearts were sectioned and stained using calcium-staining Alizarin Red S. Additional sections were also stained with Hemotoxylin and Eosin stain.</p> <p><strong>Results:</strong> From my analysis, it was apparent that there were no calcifications on the mitral valves in any of the samples. Pigmentation was apparent, but potential calcium was ruled out when compared to the H&amp;E stains. There were heavy calcium deposits in the aorta and aortic valve in forty percent of the MVD samples. Additionally, calcium was seen near the apex of the left ventricle in three of the ten MVD samples.</p> <p><strong>Conclusion and Potential Impact:</strong> In conclusion, it doesn’t appear that calcification occurs in the mitral valve, neither in the early nor late onset of RA. However, I pursued the calcifications seen in the aorta because of previous research done with calcifications seen in RA mice eating a high-fat diet. Currently, I am investigating further with the entirety of the aorta and staining to see if the calcification is isolated near the valve or if the calcifications spread. I am hoping that my findings will assist in identifying the physiological changes that are seen in rheumatoid arthritis.</p> Elena Konrath, Dr. Bryce Binstadt, M.D., Ph.D Copyright (c) 2019 Elena Konrath, Dr. Bryce Binstadt, M.D., Ph.D https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23513 Tue, 08 Oct 2019 14:18:13 -0400 Role of perilipins in the development of fatty liver disease. http://journals.iupui.edu/index.php/IMPRS/article/view/23518 <p><strong>Background and Hypothesis:</strong> 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<sup>-/-</sup> 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.</p> <p>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<sup>-/-</sup>HFD compared to WTHFD mice.</p> <p><strong>Methods:</strong> Immunostaining was used to visualize intracellular distribution of PLINs. Western blotting was used to quantify differences in PLINs protein expression. </p> <p><strong>&nbsp;Results:</strong> LD distribution showed WT regular chow (RC) = NOD2<sup>-/-</sup>RC &lt; WTHFD&lt;&lt;NOD2<sup>-/-</sup>HFD. <br> <br>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. <br> <br>PLIN1 showed a trend for decrease in both NOD2<sup>-/-</sup>HFD and WTHFD compared with RC mice. PLIN2 appears to be decreased in WTHFD and both NOD2<sup>-/-</sup> groups compared to WTRC. PLIN3 seemed to show increased expression in WTHFD but not in NOD2<sup>-/-</sup>. PLIN4 showed a trend for decreased expression in both NOD2<sup>-/-</sup>RC and NOD2<sup>-/-</sup>HFD mice compared to WTRC.</p> <p><strong>Conclusion:</strong> Despite enlarged LD size, there was no detectable increase in PLINs expression in NOD2<sup>-/-</sup>HFD compared to WTHFD. This may be due to the impact of other LDassociated proteins in the livers of NOD2<sup>-/-</sup> mice.</p> Erica Kubascik, Madison DeGoey, Dr. Gupta, Dr. Kostrominova Copyright (c) 2019 Erica Kubascik, Madison DeGoey, Dr. Gupta, Dr. Kostrominova https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23518 Tue, 08 Oct 2019 14:18:42 -0400 XPC-Related Protection Against Carcinogen-Induced Lung Adenocarcinoma is Independent of Lung Inflammation http://journals.iupui.edu/index.php/IMPRS/article/view/23520 <p><strong>Background and Hypothesis:</strong> Cigarette smoke (CS) exposure causes lung cancer, with both DNA damage and local inflammation playing a critical role in development. Our previous research links the DNA repair protein Xeroderma Pigmentosum Group C (XPC) with protection against lung cancer in CS- and carcinogen-exposed mouse models. In mice (XPC-deficient and wild-type [WT] littermates) exposed to continuous CS for 9 months, neither XPC-deficient nor WT mice develop lung cancer. XPC-deficient but not WT mice exposed to 5 months CS + 4 months air control (AC) (recovery model) develop lung cancer. In a direct carcinogen model, XPC-deficiency accelerates NTCU lung squamous cell development. Our hypothesis is that lung cancers in XPC-deficient mice are independent of treatment alterations in BAL inflammation.</p> <p><strong>Experimental Design or Project Methods:</strong> Acellular bronchoalveolar lavage (BAL) samples from the three different mouse CS and carcinogen models were analyzed for the inflammatory cytokines IL-6, IL10, IL-12p70, IL-17A, IFN-γ, MCP-1, and TNF-α by a cytometric bead array (BD Biosciences) using a flow cytometer (FACScan) according to the manufacturer’s instructions. Raw data (mean fluorescence intensity) was analyzed by BD CBA Software. Statistical comparisons were by ANOVA, with p&lt;0.05 considered significant.</p> <p><strong>Results:</strong> All measured cytokine levels were low or undetectable in BAL from AC and CS mice, with no significant XPC genotype or treatment-related changes in the measured cytokines. Differences were observed in TNF-α and IL-6 between continuous and recovery CS models independent of treatment or genotype. NTCU caused a significant increase in BAL IL-6 independent of genotype. IL-17a was elevated in NTCU-treated mice that developed lung squamous cell carcinoma.</p> <p><strong>Conclusion and Potential Impact:</strong> The XPC genotypic variation seen in lung carcinogenesis appears to be independent of BAL cytokines, suggesting that the variation is due to DNA damage rather than differences in local inflammation. Further mechanistic investigations will focus on DNA damage and repair as drivers of XPC-deficient lung cancers.</p> <p>&nbsp;</p> Isaac Lamb, Huaxin Zhou, Patricia Smith, Catherine R. Sears, M.D. Copyright (c) 2019 Isaac Lamb, Huaxin Zhou, Patricia Smith, Catherine R. Sears, M.D. https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23520 Tue, 08 Oct 2019 14:19:17 -0400 Multiscale mathematical modeling of aqueous humor dynamics: Applications for precision care in glaucoma http://journals.iupui.edu/index.php/IMPRS/article/view/23522 <p><strong>Background and Hypothesis:</strong> Reduction of intraocular pressure (IOP) resulting from overproduction or impaired outflow of aqueous humor is, to date, the only approved medical treatment for the disease. IOP is determined by the balance between aqueous humor production at the ciliary body and drainage at the anterior chamber angle. A mathematical model describing aqueous humor (AH) flow was employed in order to study the effects of glaucoma medications and risk factors on IOP.</p> <p><strong>Experimental Design or Project Methods:</strong> IOP can be calculated as the unknown variable in the mathematical balance between AH inflow (J<sub>in</sub>) and outflow (J<sub>out</sub>). Model simulations using MATLAB were used to calculate IOP in conditions of varied trabecular meshwork resistance (R) over changes in episcleral venous pressure (EVP), uveosclearal outflow facility (C<sub>uv</sub>), and ciliary capillary blood pressure (cBP) in order to simulate the effects of EVP reducing medications, prostaglandin analogs, and systemic BP, respectively.</p> <p><strong>Results: </strong>The simulated effects of EVP reducing medications and prostaglandin analogs led to IOP decrease of 13.12%, 5.51%, and 2.88% for conditions of R<sub>0</sub>, 3R<sub>0</sub>, and 6R<sub>0</sub>, respectively, where R<sub>0</sub> is equal to the trabecular meshwork resistance when IOP = EVP. The simulated effects of prostaglandin analogs resulted an IOP decrease of 13.73%, 18.34%, and 19.83%, for conditions of R<sub>0</sub>, 3R<sub>0</sub>, and 6R<sub>0</sub>, respectively. The simulated effects of increasing systemic BP resulted in an IOP increase of 49.88%, 58.51%, and 60.71% for conditions of R<sub>0</sub>, 3R<sub>0</sub>, and 6R<sub>0</sub>, respectively.</p> <p><strong>Conclusion and Potential Impact:</strong> The model simulations predict differential efficacy in IOP reducing medications in patients of varied R as well as the differential impact of systemic BP on IOP in patients of varied R. This model has the potential to predict the IOP reducing effect of medications in an individual as well as the impact of risk factors such as systemic BP on IOP.</p> Matthew Lang, Alon Harris, Alice Chandra Verticchio Vercellin, Sunu Mathew, Giovanna Guidoboni Copyright (c) 2019 Matthew Lang, Alon Harris, Alice Chandra Verticchio Vercellin, Sunu Mathew, Giovanna Guidoboni https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23522 Tue, 08 Oct 2019 14:21:00 -0400 QPCR and Western Based Fitness Index for Exercised Mice http://journals.iupui.edu/index.php/IMPRS/article/view/23525 <p>Physically fit humans exhibit resistance to infectious and sedentary lifestyle diseases, as well as cancer. This resistance is mediated by alterations in the immune microenvironment. To identify mechanisms, mouse models of exercise have been used. As it has become clear that mice have a natural desire to exercise, voluntary running wheels have become the dominant exercise method eclipsing forced exercise models. To precisely record duration and intensity of exercise, mice are often singly housed with running wheels which induces stress. This project seeks to establish a quantitative molecular and biochemical fitness index for individual mice group housed with access to voluntary running wheels to correlate with disease measures. The gastrocnemius, quadriceps, and soleus muscles were dissected from exercised mice and their controls. These muscles were subject to qPCR for TNNI1 and TNNI2 transcripts, indicators of slow and fast twitch muscles, respectively. Further, the hearts were cut into superior and inferior halves and evaluated by qPCR for the genes NPPA and ATP2A2, indicators of left ventricle thickening. Lastly, a western blot evaluated p70 S6 kinase, another marker of ventricle adaption. Voluntary wheel exercised older mice failed to increase TNNI1 mRNA in the gastrocnemius and quadriceps, indicating the absence of fiber remodeling. In contrast, increased ATP2A2 transcripts were observed in the hearts. Also, increased phosphorylated form of p70 S6 were observed in voluntary exercised mice. Together these findings suggest that voluntary exercise in a group housing setting produces cardiac adaption, but may not induce the changes in skeletal muscle reported with forced exercise. This methodology will be applied to future studies where the impact of voluntary and forced exercise on immune markers and tumor growth are studied.</p> <p>&nbsp;</p> Zachary Lee, Stephanie Vargas, Bradley Schroeder, John Foley Copyright (c) 2019 Zachary Lee, Stephanie Vargas, Bradley Schroeder, John Foley https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23525 Tue, 08 Oct 2019 14:21:32 -0400 The Effects of Propofol on the Human Blood Brain Barrier http://journals.iupui.edu/index.php/IMPRS/article/view/23526 <p><strong>Background and Hypothesis:</strong> 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.</p> <p><strong>Experimental Design or Project Methods:</strong> 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<sup></sup> assay kits. To determine the role of MMPs, a broad spectrum MMP inhibitor, GM6001, was utilized and barrier properties were monitored.</p> <p><strong>Results:</strong> 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.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> <p>&nbsp;</p> Kirsten A. Lewis, Jason M. Hughes, Scott G. Canfield Copyright (c) 2019 Kirsten A. Lewis, Jason M. Hughes, Scott G. Canfield https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23526 Tue, 08 Oct 2019 14:21:57 -0400 The Function of E6BP in Normal Human and HPV-Infected Cell Lines http://journals.iupui.edu/index.php/IMPRS/article/view/23528 <p><strong>Background and Hypothesis:</strong> High-risk human papillomavirus type-16 and -18 cause approximately 70% of cervical and 60% of oropharyngeal cancers. While the majority of HPV infections are transient and resolve on their own, some infections persist and progress to cancer. The <em>in vivo</em> and <em>in vitro</em> high-risk HPV transformation ability works to immortalize primary human keratinocytes resulting from the activities of the viral oncoproteins E6 and E7. A well-studied effect of E6 is the degradation of tumor suppressor gene p53 via binding to the ubiquitin ligase E6AP. Additionally, E6 interacts with the cellular calcium-binding protein E6BP, but the functional significance remains unknown. Here we studied the effects of E6BP knockout in normal human and HPV-infected cell lines.</p> <p><strong>Experimental Design or Project Methods:</strong> CRISPR-Cas9 system was used to knockout E6BP in HPV-human cervical cancer cell lines: HeLa (high-copy HPV18) and SiHa (low-copy HPV16) as well as human keratinocyte HaCaT (mutant p53) and p53-null lung carcinoma H1299 cell lines. Cells were transfected with a puromycin-resistant CRISPR*Cas9 control plasmid (pLentiv2) and one that carries a guide RNA against E6BP (pLentiv2-E6BP) and selected with puromycin. RFP-GFP, which does not carry a puromycin resistance, was transfected into cells to establish transfection efficiency and to serve as the control for the puromycin. After successful selection, polyclonal populations of pLentiv2 control and pLentiv2-E6BP cells were screened for E6BP protein levels. pLentiv2-E6BP cell populations with reduced E6BP protein were serial diluted to generate monoclonal cell lines. Clones were then screened E6BP protein expression, and low expression clones were further processed for DNA sequencing of the E6BP gene editing site. Additionally, the effects of E6BP knockout in Siha and HeLa cells on p53, p21, and E6AP protein expression was studied by immunoblot.</p> <p><strong>Results:</strong> Monoclonal pLentiv2-E6BP cell lines for H1299 and HaCaT cells were established and as well as polyclonal pLentiv2-E6BP HeLa and SiHa cell populations. Preliminary results indicate that E6BP knockout in HeLa and Siha cells reduces the p53 and p21 levels compared to pLentiv2 control cells. Additionally, E6AP protein expression was not affected by E6BP knockout in the polyclonal SiHa cells.</p> <p><strong>Conclusion and Potential Impact:</strong> These preliminary results suggest that E6BP levels may influence p53 and p21 levels in HPV cells. It is of particular importance to further investigate if low levels of E6BP contribute to a worse outcome during HPV induced carcinogenesis.</p> <p>&nbsp;</p> Amanda Lima, Anne Rietz, Elliot A. Androphy Copyright (c) 2019 Amanda Lima, Anne Rietz, Elliot A. Androphy https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23528 Tue, 08 Oct 2019 14:22:29 -0400 Probing the Dynamics of Neuronal Proteins In Vivo Using Non-Canonical Amino Acids Tagging http://journals.iupui.edu/index.php/IMPRS/article/view/23529 <p>Background and Hypothesis: More than 600 neurological disorders have been identified, each with varying degrees of complexity and level of molecular understanding. However, current approaches are inadequate to capture the complex progressive nature of most neurological diseases. Therefore, developing techniques capable of probing the temporal dynamics of neuronal proteins in rodents, the most commonly used experimental models, is imperative for proper understanding of mechanisms driving neurological disorders. In this project, a protein labeling technique that enables selective labeling of newly synthesized proteins <em>in vivo</em> is utilized. In this technique, the non-canonical amino acid azidohomoalanine (AHA) is injected into mice to achieve global proteome labeling. AHA is an azide-tagged methionine (Met) analog that is incorporated into the nascent proteins using endogenous translational mechanisms. The azide functional group of AHA allows selective enrichment of the newly synthesized proteins from brain tissues via click-chemistry using alkynebearing affinity tags. This will be followed by detecting the AHA-labeled protein using mass spectrometry. We hypothesize that this labeling technique will help map the dynamics of the brain proteome in health and disease. This will ultimately provide insights into mechanisms underlying complex neurological diseases.</p> <p><strong>Experimental Design or Project Methods:</strong> C57Bl/6 murine dams were injected with 0.1 mg/g AHA for two days. Brain tissues were harvested, homogenized and lysates were reacted with biotin-alkyne using copper-catalyzed click reaction. Biotinylated proteins were then enriched using NeutrAvidin beads and eluted by boiling in 2% SDS.</p> <p><strong>Results:</strong> Tissues were fractionated into different subcellular components (cytosolic, nuclear, membrane, cytoskeletal, and extracellular matrix) using buffers of different stringency. Western blot analysis of clicked tissues using Streptavidin-fluorophore indicated effective incorporation of AHA into different cellular fractions of brain tissues. Additionally, the analysis of eluted proteins revealed successful enrichment and elution of AHA-labeled proteins.</p> <p><strong>Conclusion and Potential Impact:</strong> Successful incorporation of AHA in nascent neuronal proteins can lead to a comprehensive quantitative approach for elucidating changes in the regulation of neuronal proteins in disease states.</p> Kevin P. Lin, Aya M. Saleh, Kathryn R. Jacobson, Sarah Calve, Tamara L. Kinzer-Ursem Copyright (c) 2019 Kevin P. Lin, Aya M. Saleh, Kathryn R. Jacobson, Sarah Calve, Tamara L. Kinzer-Ursem https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23529 Tue, 08 Oct 2019 14:23:23 -0400 Determining Health Information Technology Use by the Transgender Population: A Scoping Review http://journals.iupui.edu/index.php/IMPRS/article/view/23530 <p><strong>Background:</strong> Transgender people face many potential barriers in healthcare, such as real or perceived discrimination, inability to pay for healthcare services, and misinformation about transgender health needs. It has been hypothesized that, because of these barriers, transgender people may be more receptive to using health information technology than other populations. The purpose of this scoping review was to understand the ways transgender people use health information technology.</p> <p><strong>Methods:</strong> This scoping review included English studies that addressed use of technology by transgender people in health sciences literature. The inclusion criteria was studies that documented transgender technology use and did not include studies that only focused on technology use by healthcare providers. Included studies were sorted into categories based on the type of technology transgender participants used.</p> <p><strong>Results:</strong> Twenty-nine articles met the study inclusion criteria from an initial pool of 1,276 articles searched from online databases. Many studies were involved with multiple categories. Fourteen articles addressed websites targeting transgender people, twelve included the usage of online social media sites, seven articles involved transgender usage of online surveys, and four articles discussed transgender usage of smartphones in health management.</p> <p><strong>Conclusion and potential impact:</strong> Twenty-two studies focused on the application of interventions through websites and social media sites, nineteen of which concluded that web-based health information or interventions were feasible methods to affect the health of transgender people. Sixteen studies concluded that online interactions were accepted, if not preferred, by their transgender participants. This review suggests that further integration of online interventions and healthcare information into these mediums may increase transgender engagement in healthcare and reduce healthcare barriers. Future research to improve understanding of the outcomes of health information technology on the health of transgender people would be an asset for treating a historically medically underserved community.</p> Albert Liu, Joy Lee, Michael Weiner Copyright (c) 2019 Albert Liu, Joy Lee, Michael Weiner https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23530 Tue, 08 Oct 2019 14:24:01 -0400 Patient-Reported Outcomes with Tibial Baseplate Position and PCL Status in Conforming Total Knee Arthroplast http://journals.iupui.edu/index.php/IMPRS/article/view/23532 <p><strong>Background and Hypothesis:</strong> 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 <em>conforming polyethylene</em> 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.</p> <p><strong>Experimental Design or Project Methods:</strong> 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.</p> <p><strong>Results:</strong> Analyses indicated differences based on ≥8mm vs. &lt;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 &lt;8mm, PCL-preserved ≥8mm, PCL-resected &lt;8mm, and PCL-resected ≥8mm) did not differ by demographics/covariates (<em>p</em>≥0.150), tibial slope (<em>p</em>≥0.132), or preoperative PROMS (<em>p</em>≥0.088). The PCL-released &lt;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.</p> <p><strong>Conclusion and Potential Impact:</strong> In conforming polyethylene TKAs, releasing the PCL in combination with AP tibial baseplate placement &lt;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.</p> Joseph A. Madden, BS, Payton K. Arnold, MS, Evan R. Deckard, BSE, R. Michael Meneghini, MD Copyright (c) 2019 Joseph A. Madden, BS, Payton K. Arnold, MS, Evan R. Deckard, BSE, R. Michael Meneghini, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23532 Tue, 08 Oct 2019 14:24:33 -0400 Hemoglobin Expression and Function in Human Corneal Epithelium http://journals.iupui.edu/index.php/IMPRS/article/view/23565 <p>The corneal epithelium forms the outer layer of the cornea. It provides mechanical protection, prevents fluid loss and forms a barrier to invasive pathogens. Though hemoglobin expression has been extensively studied in erythroid cells, recent work in multiple cell systems has documented hemoglobin expression in cells of non-erythroid origin. However, the function of hemoglobin in non-erythroid cells has yet to be established. The hypothesis that hemoglobin is expressed in corneal epithelium and that it functions to protect against oxidative stress will be examined in the present work. Hemoglobin expression and function was examined by immunocytochemistry and western blot analysis. Native human corneal explants and an immortalized human corneal epithelial cell line were examined. Expression of hemoglobin beta and delta chains was demonstrated at the protein level in both native and cell culture preparations. Hexamethylene bisacetamide is a known inducer of hemoglobin beta chain expression in murine erythroleukemia cells. HMBA did function to increase beta chain expression in cultured corneal epithelium as well. Beta chain immunolocalization was primarily cytoplasmic, while the delta chain localized to both cytoplasmic and membrane domains. Oxidative stress, from hydrogen peroxide exposure, was shown to upregulate delta chain expression. In conclusion, hemoglobin chains are expressed in corneal epithelium and could function to protect against oxidative stress. This is relevant given that exposure to light and high oxygen tensions render the cornea particularly susceptible to oxidative damage.</p> Ryan McBride, Erin Perez, Ernest Talarico, Brian Kennedy Copyright (c) 2019 Ryan McBride, Erin Perez, Ernest Talarico, Brian Kennedy https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23565 Tue, 08 Oct 2019 14:25:54 -0400 Thrombopoietin: Role in Fracture Healing and Pain http://journals.iupui.edu/index.php/IMPRS/article/view/23537 <p><strong>Background and Hypothesis:</strong> <br>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. </p> <p><strong>Experimental Design or Project Methods:</strong> <br>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.</p> <p><strong>Results:</strong> <br>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.</p> <p><strong>Conclusion and Potential Impact:</strong> <br>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.</p> Luke G. McVeigh, Deepa Shiek Pran Babu, Rachel J. Blosser, Eric L. Thompson, Jill C. Fehrenbacher, Fletcher A. White, Melissa A. Kacena Copyright (c) 2019 Luke G. McVeigh, Deepa Shiek Pran Babu, Rachel J. Blosser, Eric L. Thompson, Jill C. Fehrenbacher, Fletcher A. White, Melissa A. Kacena https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23537 Tue, 08 Oct 2019 14:28:35 -0400 Organizational Culture Drives Confidence in Implementing Telestroke Care http://journals.iupui.edu/index.php/IMPRS/article/view/23538 <p><strong>Background and Hypothesis:</strong> During the past two years, the Veteran Affairs’ National Telestroke Program (NTSP) has conducted 800 teleconsultations at 30 participating sites. Site perceptions of the NTSP are collected in surveys at the time of the program entry and 6-12 months later. This study aims to determine factors associated with the confidence in providing stroke care among staff at participating hospitals, and whether confidence is associated with site performance on program quality indicators.</p> <p><strong>Experimental Design:</strong> Web-based surveys were sent to participating sites at baseline and post implementation. Survey questions pertained to a site’s confidence providing stroke care, organizational readiness to change (ORCA assessment), buy-in of clinical services, and the impact of the program. Individual responses were averaged at each site. Confidence was scored from zero to ten and was dichotomized into fully confident (10) and not fully confident sites (&lt;10). Site performance on key indicators of program quality were obtained from the NTSP data. A Kruskal Wallis analysis examined the association of baseline variables with postimplementation confidence.</p> <p><strong>Results:</strong> 54 individuals (57% nurses, 26% providers, and 7% administrators) from 16 sites had baseline and post-implementation data. Five sites were fully confident at the postimplementation assessment. The other 11 had confidence scores ranging from 8.32 to 9.5. Higher ORCA scores were significantly associated with post-implementation confidence (mean ORCA 12.2 vs. 6.8, p=0.04). Sites with longer NTSP participation were less likely to be fully confident, (mean 10.6 months vs. 3.8, P=0.01). Baseline confidence, rurality, and volume were not associated with post-implementation confidence, nor was post-implementation confidence associated with measurements of site performance.</p> <p><strong>Conclusion:</strong> Higher ratings of site organizational culture was associated with higher postimplementation confidence in providing stroke care. This suggests that an individual site’s context is a larger driver of the confidence in program implementation than volume or program performance. In addition, the finding that sites with longer time spent in the NTSP had lower post-implementation confidence suggests sites may benefit from periodic retraining to sustain confidence in their ability to provide stroke care.</p> Christopher Miao, Holly Martin, MPH, Teresa Damush, PhD, Michelle LaPradd, MS, Susan Ofner, MS, Linda Williams, MD Copyright (c) 2019 Christopher Miao, Holly Martin, MPH, Teresa Damush, PhD, Michelle LaPradd, MS, Susan Ofner, MS, Linda Williams, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23538 Tue, 08 Oct 2019 14:29:03 -0400 Generation of liver organoids by 3D co-culturing of hepatocytes, hepatic stellate cells, and liver sinusoidal endothelial cells http://journals.iupui.edu/index.php/IMPRS/article/view/23539 <p><strong>Background and Hypothesis:</strong> Recent progress with combination of chemicals in cell culture media prolongs hepatocyte (HC) function <em>in vitro</em>. However, without other cells of hepatic lineage that comprise natural liver, HC alone are not suitable for the study of complex liver diseases. Hepatic stellate cells (HSC) and liver sinusoidal endothelial cells (LSEC) play vital roles in physiological and pathological liver function <em>in situ</em>. We hypothesized that coculturing primary HC with HSC and LSEC as 3D liver organoids in the same chemical conditioned media would uphold HC function over time, creating a better physiological 3D liver microenvironment for liver disease research.</p> <p><strong>Experimental Design:</strong> Freshly thawed primary human HCs were combined with immortalized human HSC alone, or together with immortalized human LSECs, to generate 3D liver spheroids. Spheroids were formed in HC maintenance media (HMM, Lonza) using low adhesion 96-well plates for 6 days before switching to the media with different combination of chemicals (SB31542, Forskolin, IWP2, DAPT, and LDN193189) for culturing another 14 days. Spheroids characterization, albumin secretion, mRNA transcription (CYP3A4), and histological analysis were performed for HC differentiation, maturation, and function. </p> <p><strong>Results:</strong> Both co-cultures of HC:HSC (2.5:1 ratio), and HC:HSC:LSEC (2.5:1:1) formed spheroids in HMM within 4 to 6 days (Fig.1A). The introduction of the different chemical-based media affected the roundness and diameter of spheroids differently (Fig.1B). In the presence of all 5 chemicals (5C), HC function was better maintained up to 21 days in HC:HSC:LSEC spheroids, measured by albumin, CYP3A4, and CK-19 secretion (Fig.1C). Surprisingly, 5C-based media significantly upregulated the expression of CK-19, which is one of the markers for cholangiocytes and liver precursor cells. </p> <p><strong>Conclusion and Potential Impact:</strong> 3D liver organoids composed of HC, HSC, and LSEC would create a niche environment mimicking the<em> in vivo</em> condition. Optimization of complex hepatic spheroids, as well as optimization of complex hepatic spheroid culturing media, would allow for the generation of a unique model for studying complex liver diseases.</p> Sean C. Mulligan, MS1, Wenjun Zhang, PhD, Lester J. Smith, PhD, Erika Gramelspacher, BS, Ping Li, PhD, Burcin Ekser, MD, PhD Copyright (c) 2019 Sean C. Mulligan, MS1, Wenjun Zhang, PhD, Lester J. Smith, PhD, Erika Gramelspacher, BS, Ping Li, PhD, Burcin Ekser, MD, PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23539 Tue, 08 Oct 2019 14:29:54 -0400 Health Care System Distrust, Race, and Surrogate Decision Making Regarding Code Status http://journals.iupui.edu/index.php/IMPRS/article/view/23540 <p><strong>Background and Hypothesis:</strong> 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.</p> <p><strong>Experimental Design or Project Methods:</strong> 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.</p> <p><strong>Results:</strong> 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).</p> <p><strong>Conclusion and Potential Impact:</strong> 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. </p> Sang Yoon Na, BS, MS, James E. Slaven, MS, Emily S. Burke, BA, Alexia M. Torke, MD, MS Copyright (c) 2019 Sang Yoon Na, BS, MS, James E. Slaven, MS, Emily S. Burke, BA, Alexia M. Torke, MD, MS https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23540 Tue, 08 Oct 2019 14:30:38 -0400 GENERATION OF MOSAIC STEM CELL DERIVED INNER EAR ORGANOIDS TO DETERMINE PARACRINE EFFECTS OF TMPRSS3 http://journals.iupui.edu/index.php/IMPRS/article/view/23542 <p><strong>Background and Hypothesis:</strong> 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 Tmprsss3<em><sup>KO</sup></em> mosaic organoids would have greater hair cell survival than Tmprss3<em><sup>KO</sup></em>-only organoids due to the compensatory effect of intact TMPRSS3 on control hair cells within the same vesicle.</p> <p><strong>Experimental Design or Project Methods:</strong> Two mESC cell lines (R1E background) were previously generated using CRISPR-Cas9n. Control reporter line (tdTomato) expresses a<em> tdTomato</em> reporter gene in all cells under the CAG promoter at the ROSA26 locus. Tmprss3 knockout line (Tmprss3<em><sup>KO</sup></em>) contains a 2A-nGFP cassette with a premature stop codon into exon 2 of the <em>Tmprss3 gene</em>. We generated <em>Tmprss3<sup>KO</sup></em> mosaic (<em>Tmprss3<sup>KO</sup></em>: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.</p> <p><strong>Results:</strong> We have successfully used mESCs to generate <em>Tmprss3<sup>KO</sup></em> 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 <em>Tmprss3<sup>KO</sup></em> mosaic organoids. Additionally, the <em>Tmprss3<sup>KO</sup></em> mosaic organoids had a significantly decreased overall BK channel expression by D38. </p> <p><strong>Conclusion and Potential Impact:</strong> 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 <em>Tmprss3<sup>KO</sup></em> rescue.</p> Radoslaw Nabrzyski, BS, Pei-Ciao Tang, PhD, Alpha Alex, Rick F. Nelson, MD, PhD Copyright (c) 2019 Radoslaw Nabrzyski, BS, Pei-Ciao Tang, PhD, Alpha Alex, Rick F. Nelson, MD, PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23542 Tue, 08 Oct 2019 14:35:50 -0400 Prevalence of impairing behavioral health problems in ED patients and association with ED utilization. http://journals.iupui.edu/index.php/IMPRS/article/view/23544 <p><strong>Background and Hypothesis:</strong> <br> 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.</p> <p><strong>Experimental Design or Project Methods:</strong> <br>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-MH<sup>TM</sup>, 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.</p> <p><strong>Results:</strong> <br> 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,&nbsp;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.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> Setarah Mohammad Nader, Paul Musey Jr., MD, MS, FACEP Copyright (c) 2019 Setarah Mohammad Nader, Paul Musey Jr., MD, MS, FACEP https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23544 Tue, 08 Oct 2019 14:36:23 -0400 Targeted Elastin-like Fusion Proteins for Enhanced Imaging and Treatment of Non-Muscle Invasive Bladder Cancer http://journals.iupui.edu/index.php/IMPRS/article/view/23546 <p><strong>Background and Hypothesis:</strong> 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).</p> <p><strong>Experimental Design or Project Methods:</strong> Using synthetic biology approaches, a library of ELP fusions were made and purified using a simplified organic extraction method. (Biomaterials Science<strong> 2018</strong>, 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.</p> <p><strong>Results:</strong> 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.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> <p>&nbsp;</p> Matthew L. Nordland, Craig J. Sweet, Mollie Shinkle, David H. Thompson Copyright (c) 2019 Matthew L. Nordland, Craig J. Sweet, Mollie Shinkle, David H. Thompson https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23546 Tue, 08 Oct 2019 14:36:56 -0400 Targeted Ablation of AIMP1 in Lipopolysaccharide-induced Acute Lung Injury http://journals.iupui.edu/index.php/IMPRS/article/view/23557 <p><strong>Background and Hypothesis:</strong> <br>Infants with Bronchopulmonary dysplasia (BPD), a chronic lung disease of premature infants, are more susceptible to acute lung injury (ALI). Endothelial-Monocyte Activating Polypeptide II (EMAP II, encoded by <em>Aimp1</em>) is a proinflammatory cytokine that originates from bronchiolar club cells and plays a role in the inflammatory response during BPD development. Prolonged EMAP II exposure has been shown to worsen BPD pathogenesis by activating alveolar macrophages. The targeted ablation of EMAP II in the bronchial club cells of a mouse model (<em>Scgb1a1-ERTCre;Aimp1/fl/fl</em>) is hypothesized to decrease the inflammatory effects of ALI. </p> <p><strong>Experimental Design:</strong> <br>Aged-matched littermate mice (ages ranging from 20-25 weeks) with Tamoxifen inducible, Cremediated, bronchial club cell specific ablation of Aimp1 (<em>Scgb1a1-ERTCre;Aimp1/fl/fl</em>, denoted cKO) or with only partial ablation (Scgb1a1-ERT2Cre;Aimp1/fl/wt denoted Ctrl) were given three doses of 120 micro-liters of 20mg/ml tamoxifen over a seven day period. 24 hours after the final dose they were administered a single intratracheal delivery of lipopolysaccharide (LPS) (5mg/kg) 24 hours later immunohistochemistry (IHC) for EMAP II and inflammation was assessed by immunoblotting (IB) for IL-6 in bronchoalveolar lavage (BAL) and cytospin of BAL.</p> <p><strong>Results:</strong> <br>IHC showed a decrease of EMAP II expression in the bronchioles of the cKO as compared to ctrl. IL-6 was increased 1.95 fold in the BAL fluid of cKO by IB. Cytospin analysis showed: (Cell Type: Ctrl%, cKo%), Macrophages: 4.66%, 4.70%, Mature neutrophils: 44.44%, 60.25%, Banded Neutrophils: 41.93%, 26.20%, Lymphocytes: 4.30%, 5.10%, Eosinophils: 2.51%, 2.10%, Monocytes: 2.15%, 1.64%.</p> <p><strong>Conclusions:</strong> <br>Ablation of bronchial club cell EMAP II, in LPS-induced ALI increased the amount of IL-6 and percentage of mature neutrophils in bronchoalveolar lavage fluid. No significant difference in macrophage were noted in either group. These findings suggest that EMAP II influences immune response time; however, more experiments would be required to establish a link.</p> Chris O’Connor, Margaret Schwarz, MD, Daniel Lee, PhD Copyright (c) 2019 Chris O’Connor, Margaret Schwarz, MD, Daniel Lee, PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23557 Thu, 10 Oct 2019 10:23:20 -0400 The Effect of Chronic Hypoxia on Airway Remodeling in Murine Model of Asthma http://journals.iupui.edu/index.php/IMPRS/article/view/23560 <p><strong>Background and Hypothesis:</strong> 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% O<sub>2</sub>) exhibit lower airway reactivity, compared to room air controls (20% O<sub>2</sub>) 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. </p> <p><strong>Methods:</strong> 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.</p> <p><strong>Results:</strong> Measured ASM (mm<sup>2</sup>) 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).</p> <p><strong>Conclusions and Potential Impact:</strong> 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.</p> Jordan Ozolin, Amy Gao, Page Perez, Robert Tepper, MD PhD Copyright (c) 2019 Jordan Ozolin, Amy Gao, Page Perez, Robert Tepper, MD PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23560 Tue, 08 Oct 2019 14:37:37 -0400 The Effects on Ishikawa Endometrial Cancer Cell Lines upon Treatment with Hormonal Agents and Novel Drug Target http://journals.iupui.edu/index.php/IMPRS/article/view/23561 <p><strong>Background and Hypothesis:</strong> Endometrial cancer is the most common gynecological malignancy in the US and will claim the lives of over 12,000 women in 2019. Women with high-risk histologic tumors, such as high-grade endometrial adenocarcinoma, represent over 50% of these deaths. Novel treatments are needed to treat these aggressive tumors. Decreased expression of the ribonuclease DICER is associated with high-grade and recurrent endometrial adenocarcinoma. To study the role of DICER in endometrial cancer, we used CRISPR/Cas9 to delete DICER in an endometrial cancer cell line. RNA sequencing revealed dysregulation of steroid hormone receptor signaling and high expression of APEX1 [apurinic/apyrimidinic endonuclease 1 (Ref-1/APE1)]. Elevated levels of Ref-1/APE1 have been found in various tumor types, and Ref1/APE1 inhibitors are in clinical trials. We hypothesize that our DICER deleted cell lines will be hormone insensitive yet respond to Ref-1/APE1 inhibitors by decreasing cellular proliferation.</p> <p><strong>Project Methods:</strong> Ingenuity Pathway Analysis provided an unbiased approach to determine dysregulated genes in steroid hormone signaling in our RNA transcriptomic data. QPCR was used to confirm gene expression in independent samples. Cells were treated with steroid hormones or Ref-1/APE1 inhibitors and proliferation was assessed using MTS.</p> <p><strong>Results:</strong> DICER deleted cell lines had downregulated expression of steroid hormone receptors. Treatment with steroid hormones did not have a significant effect on proliferation. However, treatment with Ref-1/APE1 inhibitors resulted in a significant decrease in proliferation.</p> <p><strong>Conclusion and Potential Impact:</strong> Ref-1/APE1 inhibitors affect cellular proliferation of endometrial cancer cell lines. Because survival has actually worsened for women with endometrial cancer, novel therapies such as Ref1/APE1 inhibitors deserve future study. </p> Pooja Pandita, Xiyin Wang, Shannon Hawkins Copyright (c) 2019 Pooja Pandita, Xiyin Wang, Shannon Hawkins https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23561 Tue, 08 Oct 2019 14:39:20 -0400 Non-invasive estimation of prostate tumor grade with PET [68Ga] Ga-PSMA-11 imaging http://journals.iupui.edu/index.php/IMPRS/article/view/23563 <p><strong>Background and Hypothesis:</strong> In the US alone, more than 160,000 men are diagnosed with prostate cancer each year. Prostate cancer grows slowly in many patients such that most die due to unrelated causes, yet 29,000 prostate cancer-related deaths occur annually in the US. The gravity of this mortality rate promotes over-treatment of low- and intermediate-risk cancer patients as well as monitoring of cancer via invasive biopsies to determine cancer grade. We hypothesize that molecular imaging methods can differentiate low- and intermediate- risk prostate cancer, guide optimal treatment, and eliminate morbidity associated with biopsy procedures. </p> <p><strong>Experimental Design or Project Methods:</strong> In this work, we used PET [<sup>68</sup>Ga] Ga-PSMA-11 imaging to estimate cancer grade through direct comparison with whole mount pathology, which is the gold standard for cancer grading. A tool was developed using Interactive Data Language (IDL) to automate 3D spatial registration of whole mount pathology slices with [<sup>68</sup>Ga] Ga-PSMA-11 PET images. Upon accurate spatial registration, quantitative analysis was performed to establish the relationship between regional [<sup>68</sup>Ga] Ga-PSMA-11 uptake and prostate cancer grade.</p> <p><strong>Results:</strong> Thus far, we have shown qualitatively that the whole mount pathology accurately maps to the correct 3D spatial location on the PET images. Upon spatial registration, using a ROC curve, we have found that [<sup>68</sup>Ga] Ga-PSMA-11 uptake could be used to detect both low- and high- risk tumor location (AUC = 0.82 and 0.84 respectively). Further, we demonstrated increasing uptake with greater risk.</p> <p><strong>Conclusion and Potential Impact:</strong> This study begins to establish the feasibility of noninvasive, imaging-based monitoring alternatives which can mitigate the need for invasive biopsies for prostate cancer. Ultimately, improved imaging methods have the potential for not only bettering prostate cancer monitoring but also for improving cancer outcomes and quality of life for prostate cancer patients.</p> Neal Patel, Clint Bahler, Mark Green, Liang Cheng, Gary Hutchins Copyright (c) 2019 Neal Patel, Clint Bahler, Mark Green, Liang Cheng, Gary Hutchins https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23563 Tue, 08 Oct 2019 14:39:49 -0400 Effects of diabetes-related stimuli on soluble epoxide hydrolase expression http://journals.iupui.edu/index.php/IMPRS/article/view/23564 <p><strong>Background and Hypothesis:</strong> Diabetic retinopathy (DR) is a frequent complication of diabetes and is a leading cause of blindness in the developed world. While the treatment of DR has progressed with the use of anti-VEGF therapies, the underlying mechanisms for DR are still unknown. Soluble epoxide hydrolase (sEH) activity has been linked to retinal neovascularization, which can lead to blindness in proliferative DR. The use of sEH inhibitors also slowed the progression of experimental DR. In particular, sEH might be expressed in the retinal pigment epithelium (RPE), which is a critical player in metabolic processing and could be influenced by diabetes-related stimuli to promote neovascularization. We hypothesize that stimuli seen in diabetes can lead to sEH overexpression in human retinal pigmented epithelial cells (ARPE-19).</p> <p><strong>Experimental Design or Project Methods:</strong> To test our hypothesis, we examined sEH expression in ARPE-19 cells treated under hyperglycemic and hypoxic conditions similar to those of a diabetic retina. ARPE19 were treated with increasing doses of these stimuli and underwent RNA isolation, RT-PCR to study mRNA expression of sEH, and immunoblotting to study protein expression of sEH.</p> <p><strong>Results:</strong> After hyperglycemia treatment, there was a 1.5-fold increase in protein level of sEH from the control at 10 mM glucose and a 1.2-fold increase at 20 mM glucose. However, there was also a 0.5-fold decrease in sEH protein level for the 50 mM mannitol osmolarity control. These results require further replication. The hypoxia experiment is expected to show higher levels of sEH than the normoxia control and the mRNA expression analyses are underway.</p> <p><strong>Conclusion and Potential Impact:</strong> The results from this study suggest that hyperglycemic conditions can upregulate sEH expression in retinal pigment epithelial cells. Completion of this study will further our understanding of the underlying pathophysiology of DR.</p> <p>&nbsp;</p> Neeta Patwari, Bomina Park, Timothy Corson Copyright (c) 2019 Neeta Patwari, Bomina Park, Timothy Corson https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23564 Thu, 10 Oct 2019 10:26:21 -0400 Evaluating the Efficacy of Virtual Dissector Technology Implementation In Undergraduate Anatomy. http://journals.iupui.edu/index.php/IMPRS/article/view/23566 <p><strong>Background and Hypothesis:</strong> At the graduate and professional level, computer-aided instruction demonstrated a positive impact on knowledge gains when supplementing traditional teaching methods (Wilson et al). The present research sought to determine the efficacy of supplementing traditional pedagogical methods in an undergraduate anatomy lab with VH Dissector Software (VHD) on a large touchscreen terminal. </p> <p><strong>Experimental Design or Project Methods:</strong> Throughout Basic Human Anatomy (ANAT-A215), students took a pre-quiz before covering a topic followed by a post-quiz in the next class. Using VHD was the intervention, while using the traditional lab manual, models, and two prosected cadavers was the control. The mean difference between pre-test and post-test scores was calculated. Independent Samples T Tests compared the mean difference between intervention and control groups. <br> Post-course surveys were obtained. Calculations were made of frequencies for each question and Spearman Rho correlations between questions.</p> <p><br> <strong>Results:</strong> There was significant improvement in scores with the intervention (versus control) in the Spring 2019 semester (Mean<sub>intervention</sub> = 1.13, Mean<sub>control</sub> = 0.92, P &lt;0.01). In the post-course survey, 54.1% of students indicated that they would not recommend using VHD in future semesters. Among these students, 50.1% stated they did not find VHD beneficial, 34% stated VHD helped to rotate structures, and 23.4% stated VHD was a change of pace from other lab responsibilities. Among the students that recommend using VHD, 64.8% stated VHD helped to rotate structures, 51.4% stated VHD provided context for what they saw on models, and 48.9% stated that VHD was a change of pace from other lab responsibilities. A significant correlation was found between enjoying use of the VHD and perception of learning using VHD (r = 0.794, P &lt;0.01) </p> <p><strong>Conclusion and Potential Impact:</strong> These results indicate a benefit to supplementing traditional anatomy learning modalities with dissection software at the undergraduate level. Yet, while students recognize a benefit, they often recommended not using the tech in future semesters.</p> Jonah Persinger, Stacey Dunham, Polly Husmann Copyright (c) 2019 Jonah Persinger, Stacey Dunham, Polly Husmann https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23566 Tue, 08 Oct 2019 14:40:36 -0400 Effect of Biomechanical and Anatomic Characteristics on Patient-Reported Outcomes Following Total Hip Arthroplasty http://journals.iupui.edu/index.php/IMPRS/article/view/23567 <p><strong>Background and Hypothesis:</strong> The success of total hip arthroplasty (THA) is often reported in terms of infection and dislocation rates. While studies have examined the effect of acetabular cup position, anteroposterior (AP) femoral stem alignment, changes in leg length and femoral stem offset on dislocation rates, few studies have investigated the effect of these biomechanical parameters on patient-reported outcome measures (PROMS). The purpose of this study was to evaluate how PROMS may differ by THA component placement in a consecutive series of primary THAs. Awareness of the importance of implant positioning may lead to improved surgical technique and optimized PROMS.</p> <p><strong>Experimental Design or Project Methods:</strong> 933 consecutive posterolateral approach primary THAs performed between 2011 and 2018 by one surgeon were retrospectively reviewed. Acetabular cup abduction, femoral stem alignment, changes in leg length and total femoral offset were measured on APview radiographs. Prospectively collected Hip Disability and Osteoarthritis Outcome Score/HOOS Jr., University of California Los Angeles/UCLA Activity Level, and satisfaction (5-point Likert scale) were evaluated at minimum one-year.</p> <p><strong>Results:</strong> 743 THAs were analyzed. Mean age and BMI were 64 years and 31 kg/m<sup>2</sup>, respectively. After multivariate analysis, females with neutral to valgus stem placement (<em>p</em>=0.020) and patients with neutral to valgus stem placement regardless of lumbar pain (<em>p</em>=0.034) were more satisfied. In addition, patients with lumbar pain (<em>p</em>&lt;0.001) and patients with high BMI in combination with increased change in femoral offset (<em>p</em>=0.056) had lower overall HOOS Jr. scores. Interestingly, change in leg length was not a significant predictor of any PROMS (power [1-β]≥88.4%).</p> <p><strong>Conclusion and Potential Impact:</strong> AP stem alignment may play a role in increased activity level and satisfaction. In addition, high BMI in combination with increased change in femoral offset negatively influenced HOOS Jr. scores. Unsurprisingly, the presence of lumbar pain continues to negatively affect PROMS. Further research is warranted on the influence of THA component placement, spinopelvic parameters, and PROMS.</p> Lauren Pitz, BS, Braeden W. Estes, BS, Evan R. Deckard, BSE, R. Michael Meneghini, MD Copyright (c) 2019 Lauren Pitz, BS, Braeden W. Estes, BS, Evan R. Deckard, BSE, R. Michael Meneghini, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23567 Tue, 08 Oct 2019 14:41:14 -0400 Cortical growth patterns in relation to autism spectrum disorder in ages 1-2 years http://journals.iupui.edu/index.php/IMPRS/article/view/23568 <p><strong>Background and Hypothesis:</strong> Autism Spectrum Disorder (ASD) is a common neurodevelopmental disorder with a prevalence of 2.76% among children ages 3-17 in the United States<sup>1</sup>. Some studies have linked total brain volume overgrowth or gyrification changes to ASD<sup>2,3,4</sup>. However, few have attempted to relate specific growth patterns to ASD. We hypothesize that regional differences in brain growth in subjects aged 12-24 months will correlate with diagnoses from the Autism Diagnostic Observation Schedule (ADOS).</p> <p><strong>Project Methods:</strong> The subjects for this study came from the Infant Brain Imaging Study (IBIS)<sup>5</sup>. The CIVET pipeline was used to segment T1-weighted magnetic resonance images (MRIs) into surfaces using a non-linear classification method<sup>5,6,7</sup>. CIVET quality control outputs were used for validation and to select parameters for the tasks along with previous recommendations<sup>5,8</sup>. Analysis of Functional NeuroImages (AFNI) was used to convert the CIVET output format, and Connectome Workbench was used to calculate surface curvature. Using cortical reconstructions and surface curvatures from 12- and 24-month brains, anatomically-constrained Multimodal Surface Matching (aMSM) was applied to achieve point correspondence and generate individual cortical growth maps<sup>9,10</sup>.</p> <p><strong>Results:</strong> Within the IBIS database, we found 38 individuals with ASD and 121 controls with T1weighted scans at both 12 and 24-month time points. Once individual growth maps have been generated for all subjects, Permutation Analysis of Linear Models (PALM)<sup>11</sup> will be used to determine statistically significant differences in the cortical growth patterns of ASD versus control groups.</p> <p><strong>Conclusion and Potential Impact:</strong> Research on autism may benefit from longitudinal studies of growth, as opposed to analysis of structural differences at later ages<sup>4</sup>. We concentrate on cortical growth before 24 months, which may serve as an earlier marker of ASD, when abnormal brain growth can be seen yet social deficits are not fully established<sup>5</sup>.</p> <p>&nbsp;</p> <p>[1] Zablotsky B, Black LI, Blumberg SJ. Estimated Prevalence of Children With Diagnosed Developmental Disabilities in the United States, 2014–2016. NCHS Data Brief 2017. https://www.cdc.gov/nchs/data/databriefs/db291.pdf (accessed April 29, 2019).</p> <p>[2] Libero LE, Schaer M, Li DD, Amaral DG, Nordahl CW. A Longitudinal Study of Local Gyrification Index in Young Boys With Autism Spectrum Disorder. Cereb Cortex. 2019;29(6):2575-87.</p> <p>[3] Raznahan A, Toro R, Daly E, Robertson D, Murphy C, Deeley Q, et al. Cortical anatomy in autism spectrum disorder: an in vivo MRI study on the effect of age. Cereb Cortex. 2010;20(6):1332-40.</p> <p>[4] Duret P, Samson F, Pinsard B, Barbeau EB, Bore A, Soulieres I, et al. Gyrification changes are related to cognitive strengths in autism. Neuroimage Clin. 2018;20:415-23.</p> <p>[5] Hazlett HC, Gu H, Munsell BC, Kim SH, Styner M, Wolff JJ, et al. Early brain development in infants at high risk for autism spectrum disorder. Nature. 2017;542(7641):348-51.</p> <p>[6] Shaw P, Malek M, Watson B, Sharp W, Evans A, Greenstein D. Development of cortical surface area and gyrification in attentiondeficit/hyperactivity disorder. Biol Psychiatry. 2012;72(3):191-7.</p> <p>[7] Ad-Dab’bagh, Y., Einarson, D., Lyttelton, O., Muehlboeck, J.-S., Mok, K., Ivanov, O., Vincent, R.D., Lepage, C., Lerch, J., Fombonne, E., and Evans, A.C. (2006). The CIVET Image-Processing Environment: A Fully Automated Comprehensive Pipeline for Anatomical Neuroimaging Research. In Proceedings of the 12th Annual Meeting of the Organization for Human Brain Mapping, M. Corbetta, ed. (Florence, Italy, NeuroImage). http://www.bic.mni.mcgill.ca/users/yaddab/Yasser-HBM2006-Poster.pdf</p> <p>[8] Shaw P, Kabani NJ, Lerch JP, Eckstrand K, Lenroot R, Gogtay N, et al. Neurodevelopmental trajectories of the human cerebral cortex. J Neurosci. 2008;28(14):3586-94.</p> <p>[9] Garcia KE, Robinson EC, Alexopoulos D, Dierker DL, Glasser MF, Coalson TS, et al. Dynamic patterns of cortical expansion during folding of the preterm human brain. Proc Natl Acad Sci U S A. 2018;115(12):3156-61.</p> <p>[10] Robinson EC, Garcia K, Glasser MF, Chen Z, Coalson TS, Makropoulos A, et al. Multimodal surface matching with higher-order smoothness constraints. Neuroimage. 2018;167:453-65.</p> <p>[11] Winkler AM, Ridgway GR, Webster MA, Smith SM, Nichols TE. Permutation inference for the general linear model. NeuroImage, 2014;92:381-397 (Open Access)</p> Ryan Plunkett, Emily Iannopollo, Chris Basinski, Kara Garcia Copyright (c) 2019 Ryan Plunkett, Emily Iannopollo, Chris Basinski, Kara Garcia https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23568 Tue, 08 Oct 2019 14:41:46 -0400 Association Between IL6R Polymorphisms and Cachexia Phenotype in Patients with Pancreatic Ductal Adenocarcinoma http://journals.iupui.edu/index.php/IMPRS/article/view/23710 <p><strong>Background:</strong> Cachexia, manifested as progressive adipose and muscle wasting, affects up to 80% of patients with pancreatic ductal adenocarcinoma (PDAC) and significantly increases morbidity and mortality. Increased inflammation is an underlying mechanism in almost all cases of cachexia. Trans-signaling of Interleukin-6 (IL-6) via the soluble form of its receptor (sIL6R) has been shown to promote inflammation. Certain polymorphisms of the IL6R gene such as rs2228145 (Asp358Ala substitution) are associated with increased levels of sIL6R. We hypothesize that patients with PDAC possessing IL6R alleles correlated with higher levels of circulating sIL6R will have increased systematic inflammation manifested as increased cachexia prevalence or severity.</p> <p><br><strong>Methods:</strong> DNA was extracted from prospectively collected blood samples acquired from patients with PDAC and from non-cancer controls. Genotype at the rs2228145 polymorphism was determined by TaqMan qPCR genotyping. The resulting genotypes (A/A, A/C, C/C) were compared against cachexia-related metrics, including presence of cachexia (&gt;5% body weight loss in the prior 6-months), body mass index (BMI), BMI-adjusted weight loss grade (BMI-WLG), and muscle and adipose volumes calculated from height-adjusted surface areas obtained from CT scans at the level of the third lumbar vertebra.</p> <p><br><strong>Results:</strong> 83.3% of patients with PDAC heterozygous (A/C) and 84.6% of the patients homozygous for the rs2228145 polymorphism (C/C) exhibited cachexia, versus 57.9% of patients homozygous for the reference allele (A/A), (P = 0.0364, Chi-square test). No significant difference was found among genotypes for BMI, 6-month weight loss, BMI-WL grade, or muscle and adipose tissue indices.</p> <p><br><strong>Conclusion:</strong> Patients with PDAC who are possess at least one copy of the rs2228145 polymorphism have a higher incidence of cachexia than those who are homozygous for the reference allele. This association suggests a causal role for sIL6R in cancer cachexia.</p> Nicholas J. Polster, Joseph E. Rupert, Andrew R. Young, Teresa A. Zimmers Copyright (c) 2019 Nicholas J. Polster, Joseph E. Rupert, Andrew R. Young, Teresa A. Zimmers https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23710 Tue, 29 Oct 2019 14:49:50 -0400 A Rare Case of Unilateral Foix-Chavany-Marie Syndrome http://journals.iupui.edu/index.php/IMPRS/article/view/23571 <p><strong>Background:</strong> Bilateral anterior opercular syndrome, or Foix-Chavany-Marie Syndrome (FCMS), is a neurological condition characterized by bilateral anterior opercular lesions. Common presenting symptoms of this rare suprabublar palsy include dysarthria, or slurred speech, as well as paralysis of facial, pharyngeal, lingual, and masticatory voluntary muscles while autonomic function is preserved.<sup>1</sup> Most cases of FCMS are bilateral, yet some rare cases have been reported with unilateral opercular lesions with preexisting contralateral white matter lesions.<sup>2</sup> In this case report we present a rare case of unilateral FCMS in a patient who had an acute anterior left opercular infarct, as well as a residual right parietal subcortical stroke.</p> <p><strong>Project Methods:</strong> The patient we present is a 49 year old African American female with a history of hypertension and previous stroke with residual spastic right hemiplegia who presented to the ED for evaluation of right sided facial droop, right sided weakness, and slurred speech. Patient was evaluated by acute stroke team and was treated with intravenous thrombolysis (Alteplase). On hospital day 2, the patient developed complete disarticulation, unable to produce any speech but able to communicate by appropriate gestures and writing. The patient’s symptoms included anarthria, weakness of bilateral masseters, and lateral/medial pterygoids without dysphagia. <br> <br> <strong>Results:</strong> The MRI brain showed left insular region/frontal opercular ischemic stroke and a small right parietal subcortical ischemic stroke, likely embolic in nature. Over the hospital stay, the patient’s motor function improved but her anarthria persisted. These radiologic findings along with the symptomology proved consistent with FCMS. She was discharged to home with an NIH stroke scale of 5 and recommended outpatient speech therapy.</p> <p><strong>Conclusion:</strong> In this case report, we describe a patient that presents with an extremely rare case of unilateral FCMS, with a preexisting contralateral parietal infarct that could have collectively caused anarthria and masseter weakness.</p> <p>&nbsp;</p> <p><strong>Works Cited</strong> <br>1. Milanlioglu A, Aydın MN, Gökgül A, Hamamcı M, Erkuzu MA, Tombul T. Ischemic Bilateral Opercular Syndrome. Case Reports in Medicine. 2013;2013:1-3. doi:10.1155/2013/513572.</p> <p>2. Sa F, Cordeiro IM, Mestre S, Nzwalo H. Unilateral opercular infarction presenting with FoixChavany-Marie syndrome. Case Reports. 2014;2014. doi:10.1136/bcr-2014-206439.</p> Courtney Raab, Farrukh Chaudhry, MD Copyright (c) 2019 Courtney Raab, Farrukh Chaudhry, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23571 Tue, 08 Oct 2019 14:44:33 -0400 Liver Dysfunction may be Associated with QOL in Adults with CF http://journals.iupui.edu/index.php/IMPRS/article/view/23572 <p><strong>Background:</strong> More individuals with cystic fibrosis (CF) are living into their adulthood due to significant advances in medical care. The adult CF clinic at Indiana University cares for one of the largest adult populations with CF. Liver disease is common in children and adults with CF. However, the characteristics and consequences of CF liver disease (CFLD) in adults are not well understood. In this ongoing, IRB approved study, we systematically characterized the liver manifestations and their health related quality of life in adults with well characterized CF.</p> <p><strong>Methods:</strong> Individual patient demographics and clinical data were collected for study participants. Participants completed two quality of life study questionnaires: Chronic Liver Disease Questionnaire (CLDQ) and Revised Cystic Fibrosis Questionnaire (CFQ-R). 10mL of blood was collected for future research. Finally, participants underwent a bedside transient elastography via FibroScan®.</p> <p><strong>Results:</strong> There were 50 patients, with a mean age of 30.9 years, enrolled in the study. Pancreatic insufficiency was the most common co-morbidity, as it affected 90% of patients. The mean AST was 23.55 units/L, mean ALT was 25.20 units/L, and mean alkaline phosphatase level was 117.73 units/L. From the FibroScan®, the median liver stiffness measurement (LSM) was 4.65 kPa and the mean controlled attenuation parameter (CAP) was 219.52 dB/M. The mean CLDQ score was 5.343.</p> <p><strong>Conclusion:</strong> This ongoing study reveals that increased alkaline phosphatase and CAP scores are associated with a poor QOL. More data is needed to further understand the pathophysiology behind CFLD. Use of noninvasive imaging, noninvasive markers, and the CLDQ may aid in early identification of adult CFLD.</p> <p>&nbsp;</p> <p>&nbsp;</p> Megana Rao, BS, Regina Weber, BS, Naga Chalasani, MD Copyright (c) 2019 Megana Rao, BS, Regina Weber, BS, Naga Chalasani, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23572 Tue, 08 Oct 2019 14:45:01 -0400 Epidemiological Assessment of GI BioFire Negative Tests at Parkview Health http://journals.iupui.edu/index.php/IMPRS/article/view/23573 <p><strong>Background and Hypothesis:</strong> 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.</p> <p><strong>Design:</strong> 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.</p> <p><strong>Results:</strong> 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.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> <p>&nbsp;</p> Brycen Ratcliffe, B.S., Candace Rodgers, R.N., R. Scott Stienecker, M.D. Copyright (c) 2019 Brycen Ratcliffe, B.S., Candace Rodgers, R.N., R. Scott Stienecker, M.D. https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23573 Tue, 08 Oct 2019 15:02:45 -0400 Retinal and choroidal vascular abnormalities contribute to the conversion from dry to wet age-related macular degeneration: a theoretical approach http://journals.iupui.edu/index.php/IMPRS/article/view/23574 <p><strong>Background and Hypothesis:</strong> 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.</p> <p><strong>Experimental Design or Project Methods:</strong> 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 O<sub>2</sub> supply, consumption and diffusion in: physiological baseline conditions; AMD conditions.</p> <p><strong>Results:</strong> 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.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> <p>&nbsp;</p> Lucas Rowe, Alon Harris, Thomas Ciulla, Greta Chiaravialli, Alice Chandra Verticchio Vercellin, Brent Siesky, Giovanna Guidoboni Copyright (c) 2019 Lucas Rowe, Alon Harris, Thomas Ciulla, Greta Chiaravialli, Alice Chandra Verticchio Vercellin, Brent Siesky, Giovanna Guidoboni https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23574 Tue, 08 Oct 2019 15:08:17 -0400 Chamas for Change: Redevelopment of Community Health Volunteer Curriculum for a Group-Based Maternal-Child Peer Support and Microfinance Care Model in Western Kenya http://journals.iupui.edu/index.php/IMPRS/article/view/23575 <p><strong>Background:</strong> Maternal mortality continues to be the leading cause of death for women of child-bearing age in Kenya. <em>Chamas</em> 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, <em>Chamas</em> 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 <em>Chamas</em> for Change team is looking to expand the scope of <em>Chamas</em> to additional counties in Kenya and beyond.</p> <p><strong>Project:</strong> To prepare for this transition to scale, the <em>Chamas</em> 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 <em>Chamas</em> members was analyzed. Lastly, additional topics were gathered from existing child development and adolescent reproductive health curricula through AMPATH.</p> <p><strong>Results:</strong> 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.</p> <p><strong>Conclusions and Impact:</strong> This refined flipchart will help to address demands from <em>Chama</em> mothers who are seeking a more robust education, while also further standardizing and broadening the curriculum for CHVs. It will additionally</p> Grace Rushton, BS, Julia Jerono Songok, MMBchB, Anjellah Jumah, Justus E. Ikemeri, BSc, Laura Ruhl, MD MPH Copyright (c) 2019 Grace Rushton, BS, Julia Jerono Songok, MMBchB, Anjellah Jumah, Justus E. Ikemeri, BSc, Laura Ruhl, MD MPH https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23575 Tue, 08 Oct 2019 15:08:49 -0400 SHROOM3 is a novel component of the planar cell polarity pathway whose disruption causes congenital heart disease http://journals.iupui.edu/index.php/IMPRS/article/view/23577 <p>Congenital heart disease (CHD) is the most common cause of death due to birth defects. Despite CHD frequency, the etiology remains mostly unknown. Understanding CHD genetics and elucidating disease mechanism will help establish prognosis, identify comorbidity risks, and develop targeted therapies. CHD often results from disrupted cytoarchitecture and signaling pathways. We have identified a novel CHD candidate SHROOM3, a protein associated with the actin cytoskeleton and the Wnt/Planar Cell Polarity (PCP) signaling pathway. SHROOM3 induces actomyosin constriction within the apical side of cells and is implicated in neural tube defects and chronic renal failure in humans. A recent study demonstrated that SHROOM3 interacts with Dishevelled2 (DVL2), a component of the PCP signaling pathway, <em>suggesting that SHROOM3 serves as an important link between acto-myosin constriction and PCP signaling</em>. PCP signaling establishes cell polarity required for multiple developmental processes, and is required for cardiac development. In Preliminary data we utilized a Shroom3 gene-trap mouse (Shroom3<sup>gt/gt</sup>) to demonstrated that SHROOM3 disruption leads to cardiac defects phenocopy PCP disruption. We also demonstrate that patients with CHD phenotypes have rare and potentially damaging <em>SHROOM3 variants</em> within SHROOM3’s PCP-binding domain. We hypothesize SHROOM3 is a novel terminal effector of PCP signaling, and disruption is a novel contributor to CHD. To test this, we assessed genetic interaction between SHROOM3 and PCP during cardiac development and the ultimate effect on cell structure and movement. Heterozygous Shroom<sup>3+/gt</sup> mice and heterozygous Dvl2<sup> +/-</sup> mice are phenotypically normal. We demonstrated genetic interaction between SHROOM3 and PCP signaling by generating compound heterozygous Shroom3<sup>+/gt</sup> ;Dvl2<sup> +/-</sup> mice and identifying a Double Outlet Right Ventricle and Ventricular Septal Defect in one embryo. We also observed fewer compound heterozygous mice than anticipated by Mendelian rations (observed: 18.4%; expected: 25%; n=76), suggesting potential lethality in utero. Immunohistochemistry demonstrates disrupted actomyosin in the SHROOM3<sup>gt/gt</sup> mice, characteristic of PCP disruption. These data help strengthen SHROOM3 as a novel CHD candidate gene and a component of the PCP Signaling pathway. Further characterization of this gene is important for CHD diagnosis and therapeutic development.</p> Alison Schmidt, Matthew Durbin, MS MD, James O’Kane, MS, Stephanie M. Ware, MD PHD Copyright (c) 2019 Alison Schmidt, Matthew Durbin, MS MD, James O’Kane, MS, Stephanie M. Ware, MD PHD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23577 Thu, 10 Oct 2019 10:28:03 -0400 Risk Factors for Unplanned Admission to the Pediatric Intensive Care Unit in Pediatric Trauma Patients http://journals.iupui.edu/index.php/IMPRS/article/view/23579 <p><strong>Background and Hypothesis:</strong> A number of risk factors for unplanned pediatric intensive care unit (PICU) admission and readmission have been identified. However, little is known of the risk factors associated with unplanned admission to the PICU in pediatric trauma populations. We hypothesize specific risk factors can be identified which may be associated with unplanned admission to the PICU following traumatic injury.</p> <p><strong>Methods:</strong> For this national retrospective study, we queried the 2016 National Trauma Data Bank for patients younger than 18 years experiencing a traumatic injury requiring hospital admission from the Emergency Department (ED). We excluded patients who had experienced burn injuries. Statistically significant (p&lt;.05) risk factors for unplanned PICU admission were identified in bivariate analysis and used to build a multiple logistic regression model.</p> <p><strong>Results:</strong> Patients experiencing unplanned admission to the PICU had lower ED Glasgow Coma Scale (11.83 vs. 14.31; p&lt;0.001), higher Injury Severity Scores (ISS) (17.96 vs. 7.32; p&lt;0.001), and were older (age 11.35 vs. 9.60; p&lt;0.001). Initial ED disposition to the PICU was significantly associated with unplanned admission to the PICU (p&lt;0.001). Initial ED disposition to the OR was significantly associated with unplanned admission to the PICU (p=.018). After multiple logistic regression, ISS (p&lt;0.001), initial ED disposition to the PICU (p=0.002), initial ED disposition to the OR (p=0.005), and older age (p=0.005) remained statistically significant risk factors for unplanned admission to the PICU.</p> <p><strong>Conclusion:</strong> ISS and ED disposition to the PICU, OR disposition to the PICU and age are significant risk factors for unplanned admission to the PICU in pediatric trauma. These findings will assist in identifying patients at risk for unplanned admission to the PICU, thereby reducing the adverse effects of unplanned PICU admission and ultimately improving the quality of care for pediatric trauma populations.</p> William Schrock, Jodi Raymond, MPH, Matthew Landman, MD, MPH Copyright (c) 2019 William Schrock, Jodi Raymond, MPH, Matthew Landman, MD, MPH https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23579 Tue, 08 Oct 2019 15:09:28 -0400 Tranexamic Acid Increases Early Postoperative Pain and Decreases Time to First Opioid Following Total Knee Arthroplasty http://journals.iupui.edu/index.php/IMPRS/article/view/23580 <p><strong>Background and Hypothesis:</strong> 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.</p> <p><strong>Project Methods:</strong> 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.</p> <p><strong>Results: </strong>The sex (<em>p</em>=0.393), age (<em>p</em>=0.784), and BMI (<em>p</em>=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&lt;0.001). <br>&nbsp;</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> <p>&nbsp;</p> Sachin Seetharam, Sydney Keller, Mary Ziemba-Davis, R. Michael Meneghini MD Copyright (c) 2019 Sachin Seetharam, Sydney Keller, Mary Ziemba-Davis, R. Michael Meneghini MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23580 Tue, 08 Oct 2019 15:10:47 -0400 Telepresence And Provider Communication Predict Satisfaction With Telestroke http://journals.iupui.edu/index.php/IMPRS/article/view/23581 <p><strong>Background:</strong> 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.</p> <p><strong>Methods:</strong> 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. &lt; 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.</p> <p><strong>Results:</strong> 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 &lt; 0.0001), telepresence (p &lt; 0.0001), provider communication ratings (p &lt; 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.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> Griffin Tyler Selch, Michael J. Lyerly, Holly Martin, Glenn Graham, Sharyl Martini, Jane Anderson, Teresa Damush, Michelle LaPradd, Susan Ofner, Linda Williams Copyright (c) 2019 Griffin Selch Selch, Michael J. Lyerly, Holly Martin, Glenn Graham, Sharyl Martini, Jane Anderson, Teresa Damush, Michelle LaPradd, Susan Ofner, Linda Williams https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23581 Tue, 08 Oct 2019 00:00:00 -0400 Augmenting nab-paclitaxel/gemcitabine standard chemotherapy response by merestinib, an inhibitor of c-MET, Axl and DDR signaling pathways, in preclinical pancreatic cancer models http://journals.iupui.edu/index.php/IMPRS/article/view/23583 <p><strong>Background and Hypothesis:</strong> Pancreatic ductal adenocarcinoma (PDAC) is among the most lethal malignancies in Western countries. <em>Nab</em>-paclitaxel (NPT) plus gemcitabine (Gem) is the standard of care for PDAC leading to a dismal 8.5 months median survival. Aberrant signaling of c-MET, Axl and DDR have been reported in a variety of human cancers including PDAC. Merestinib (Mer) is a potent, smallmolecule inhibitor of these pathways. We evaluated the therapeutic efficacy of merestinib to enhance the antitumor response of standard chemotherapy in preclinical models of PDAC.</p> <p><strong>Project Methods:</strong> Cell proliferation of PDAC-associated cells (AsPC-1, PANC-1 and fibroblasts) were evaluated by colorimetric WST-1 assay. Protein expression was determined by Western blot analysis. Tumor progression studies were performed in NOD/SCID mice.</p> <p><strong>Results:</strong> <em>In vitro</em> studies demonstrated that both nab-paclitaxel plus gemcitabine and merestinib suppressed cell proliferation of PDAC epithelial cells and stromal cells. Importantly, the combination treatment demonstrated additive inhibitory effects. In AsPC-1 cells, at the medium dose level, NPT+Gem, Mer and NPT+Gem+Mer treatments inhibited cell proliferation by 53.9%, 13.5%, and 81.61%, respectively. In PANC-1 cells, at the highest dose level, inhibition in cell proliferation by NPT+Gem, Mer and NPT+Gem+Mer treatments was 53.6%, 3.7%, and 72.8%. In the PDAC-associated fibroblasts, at the medium dose level, NPT+Gem, Mer and NPT+Gem+Mer treatments inhibited growth by 55.3%, 58.0%, and 91.6%. Immunoblot analysis revealed that merestinib caused a decrease in the PI-3K-AKT signaling proteins and an increase in apoptosisrelated proteins cleaved PARP-1 or cleaved caspase-3 in PDAC cells either alone or in combination with nab-paclitaxel and gemcitabine. <em>In vivo</em> study to evaluate tumor growth inhibition effects of merestinib in a subcutaneous PDAC xenograft is currently ongoing.</p> <p><strong>Conclusion:</strong> The antitumor effect of standard chemotherapy regimen can be significantly enhanced by the cMET/Axl/DDR pathway inhibitor merestinib, which may lead to clinically relevant therapeutic strategy to increased survival in PDAC patients.</p> Eda Shi Copyright (c) 2019 Eda Shi https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23583 Tue, 08 Oct 2019 15:12:05 -0400 Bone-muscle crosstalk between osteocytes and skeletal muscle through microRNA21 removal leads to sex dependent increase in muscle mass http://journals.iupui.edu/index.php/IMPRS/article/view/23584 <p><strong>Background and Hypothesis:</strong> Previous studies suggested that microRNA removal from osteocytes, the cells embedded in the bone matrix, not only has effects on bone, but also increases muscle mass only in female mice. This suggested that changes in bone cells alters skeletal muscle through bone-muscle crosstalk. We therefore tested the hypothesis that osteocytic microRNA21 removal leads to release of factors that can influence skeletal muscle.</p> <p><strong>Experimental Design or Project Methods:</strong> Osteocytic microRNA21-deficient mice were generated by mating microRNA21<sup>flox/flox</sup> mice with mice expressing Cre recombinase in osteocytes. Lean body mass was measured in 4-month-old male and female mice by DXA/Piximus. Femurs/tibias were isolated and cultured in the presence of growing media for 48h. Media was collected (CM) and transferred to differentiating C2C12 myoblastic cells. After 48h, cells were fixed and stained for myosin heavy chain. Myotube diameter was measured using ImageJ software. RNA was extracted from gastrocnemius muscle using Trizol, and gene expression was measured by qPCR. Statistical analysis was performed by t-test within each sex.</p> <p><strong>Results:</strong> Percent lean body mass was 3.33% higher in female microRNA21 knockouts compared to female microRNA21<sup>flox/flox</sup> (control) mice, with no changes observed in males. Further, CM from female knockout mice increased the mean myotube diameter by 24.3% compared to control females. However, we did not find any differences in the levels of expression of skeletal muscle genes in females or males.</p> <p><strong>Conclusion and Potential Impact: </strong>We conclude that microRNA21 deletion from osteocytes in female mice results in the release of factor(s) with anabolic or anticatabolic effects in skeletal muscle. As we determined that osteocytes must be releasing factors that influences muscle growth, future studies can be directed at specifically finding which osteocyte cytokines and/or genes are directly involved in muscle homeostasis, and whether microRNA deletion also affects skeletal muscle function/strength.</p> Andrew Sickbert, Dr. Lillian Plotkin Copyright (c) 2019 Andrew Sickbert, Dr. Lillian Plotkin https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23584 Wed, 09 Oct 2019 09:09:31 -0400 Role of CaMKK2 in Osteoclast Development and Function http://journals.iupui.edu/index.php/IMPRS/article/view/23586 <p><strong>Background and Hypothesis:</strong> In bone remodeling there are two opposing forces, osteoclast resorption and osteoblast synthesis of bone. When there is an imbalance, osteoporosis is likely found. We look to determine the role that Ca<sup>2+</sup>/calmodulin (CaM)-dependent protein kinase kinase 2 (CaMKK2) has on osteoclast development and attachment. Previous studies show that <em>Camkk2<sup>-/-</sup></em> mice have fewer mature osteoclasts that display poor attachment to the bone. These results suggest that there is a defect in facilitating osteoclast differentiation, or a defect in the attachment itself. Thus, we hypothesize that CaMKK2 plays an integral role in the development and function of the osteoclast cytoskeleton.</p> <p><strong>Experimental Design or Project Methods:</strong> We plated different concentrations of bone marrow cells isolated from male and female, wildtype and Camkk2-/- mice to investigate size and number of mature osteoclasts present. We examined the size of the resorption pits made by these osteoclasts, when plated on calcium phosphate-coated surfaces. We analyzed the RNA and protein and stained the cells immunohistochemically for proteins associated with cell attachment.</p> <p>Results: There were greater numbers of mature osteoclasts in the Camkk2-/- males at each density than the wildtype males. The female mice followed the same trend, but with fewer osteoclasts overall.</p> <p>The average area per resorbed pit was two-times larger in the WT males than the <em>Camkk2<sup>-/-</sup></em> males. But, for the females the opposite trend is shown, the pits from the Camkk2-/- females were twice as large as those formed from the wildtype females.</p> <p>To gauge efficiency, we calculated the average area resorbed per osteoclast. For the males, significantly greater efficiency from the wildtype osteoclasts was found than the <em>Camkk2<sup>-/-</sup></em> osteoclasts. For the females, there was no difference in efficiency at the lower plating densities, but the osteoclasts from the <em>Camkk2<sup>-/-</sup></em> females had greater efficiency at the greater densities.</p> <p>Remaining results are to be finalized.&nbsp;</p> <p><strong>Conclusion and Potential Impact:</strong> With the decrease in efficiency of <em>Camkk2<sup>-/-</sup></em> mice osteoclasts, we believe that CaMKK2 is necessary for the attachment of OCs. CaMKK2 could be a target to slow the pathologic breakdown of bone.</p> Maddie Smith, Uma Sankar Copyright (c) 2019 Maddie Smith, Uma Sankar https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23586 Wed, 09 Oct 2019 09:10:48 -0400 Gpr17 signaling decreases GLP-1 secretion in GLUTag cells http://journals.iupui.edu/index.php/IMPRS/article/view/23587 <p><strong>Background and Hypothesis:</strong> 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.</p> <p><strong>Experimental Design or Project Methods:</strong> 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.</p> <p><strong>Results:</strong> 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.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> Katherine Stalbaum, Shijun Yan, Hongxia Ren Copyright (c) 2019 Katherine Stalbaum, Shijun Yan, Hongxia Ren https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23587 Wed, 09 Oct 2019 00:00:00 -0400 Inhibition of P65 and PDK2 Increases Cytotoxicity and Radiation Sensitivity of Pancreatic Cancer Cells http://journals.iupui.edu/index.php/IMPRS/article/view/23589 <p><strong>Background:</strong> 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.</p> <p><strong>Experimental Design:</strong> 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</p> <p><strong>Results:</strong> 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&lt; 0.001,<em> t</em> test)</p> <p><strong>Conclusion:</strong> 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.</p> Joshua A. Streveler, Helen Chin-Sinex, Marc S. Mendonca Copyright (c) 2019 Joshua A. Streveler, Helen Chin-Sinex, Marc S. Mendonca https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23589 Wed, 09 Oct 2019 09:14:15 -0400 Type 2 Diabetes-Driven Alterations in Bone Healing and Angiogenesis http://journals.iupui.edu/index.php/IMPRS/article/view/23604 <p><strong>Background and Hypothesis:</strong> 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.</p> <p><strong>Project Methods:</strong> Using <em>Tie2-CreER;Td-Tomato</em> mice (<em>Tie2<sup>CreERT+</sup></em>), 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 <em>in vitro</em>.</p> <p><strong>Results:</strong> 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, <em>in vitro</em> 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.</p> <p><strong>Conclusion and Potential Impact:</strong> 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<em> in vitro</em> vs.<em> in vivo</em>. 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.</p> Seungyup Sun, Fazal Ur Rehman Bhatti, Ushashi C. Dadwal, Deepa Sheik Pran Babu, Anthony J. Perugini III, Olatundun D. Awosanya, Rachel J. Blosser, Sarah A. Tersey, Kara S. Orr, Karishma R. Randhave, Jiliang Li, Mervin C. Yoder, Carmella Evans-Molina, Melissa A. Kacena Copyright (c) 2019 Seungyup Sun, Fazal Ur Rehman Bhatti, Ushashi C. Dadwal, Deepa Sheik Pran Babu, Anthony J. Perugini III, Olatundun D. Awosanya, Rachel J. Blosser, Sarah A. Tersey, Kara S. Orr, Karishma R. Randhave, Jiliang Li, Mervin C. Yoder, Carmella Evans-Molina, Melissa A. Kacena https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23604 Wed, 09 Oct 2019 09:14:43 -0400 Synthetic lethality with cisplatin by targeting FACT complex in breast cancer http://journals.iupui.edu/index.php/IMPRS/article/view/23605 <p><strong>Background and Hypothesis:</strong> Previous work by our lab has shown that FACT (FAcilitates Chromatin Transcription) is upregulated from P(1º) ➝I(immortalized)➝R(Ras-transformed) cells. The FACT complex is targeted by curaxins, small molecules that “trap” FACT on chromatin, decreasing its ability to destabilize histone tetramers and facilitate DNA replication and transcription. The lab has also observed selective sensitivity to curaxins in the same pattern, P➝I➝R, suggesting the dependency of Ras-transformed cells on FACT for survival. Therefore, we hypothesized that by impairing the cancer cell’s ability to repair DNA, curaxins sensitize breast cancer cells to DNA damage-inducing chemotherapeutics, such as cisplatin. Cisplatin represents the current standard of care for triple negative breast cancer (TNBC), but is associated with exceedingly toxic side effects as well as resistance. Combination therapies may permit cisplatin dose reduction while simultaneously improving response.</p> <p><strong>Project Methods:</strong> We first determined which cancer cell lines (MCF7: ER+/PR+/HER2-; T47D: ER+/PR+/HER2-; and MDA-MB-453: TNBC) would be appropriate for demonstrating synergistic cisplatin + CBL0137 (specific curaxin) cytotoxicity. We then used BrdU assay and Western blot to assess the effects of the drugs individually and in combination on cancer cells.</p> <p><strong>Results:</strong> We observed heterogeneity in sensitivity of cancer cell lines to CBL0137 as well as statistically significant synthetic lethality between cisplatin and CBL0137.</p> <p><strong>Conclusion and Potential Impact: </strong>The knowledge gained by this research is actively being used to develop chemoprevention strategies and to enhance the effectiveness of chemotherapy. Prompted by our results, in vivo preclinical and clinical studies using cisplatin + CBL0137 to treat TNBC are planned.</p> <p>&nbsp;</p> <p>&nbsp;</p> Sarah Swiezy, Rebecca Dirks, MD, Harikrishna Nakshatri, PhD Copyright (c) 2019 Sarah Swiezy, Rebecca Dirks, MD, Harikrishna Nakshatri, PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23605 Wed, 09 Oct 2019 09:15:14 -0400 Rapid Identification of Large Vessel Occlusion Stroke Subtype in the PreHospital Setting http://journals.iupui.edu/index.php/IMPRS/article/view/23610 <p><strong>Background and Hypothesis:</strong> Stroke treatment is highly time-sensitive, with an estimated 1.9 million neurons dying per minute during an untreated ischemic stroke. The recent advent of mechanical thrombectomy (MT) and its illustrated safety and efficacy in treating large vessel occlusion (LVO) strokes has generated a need to rapidly identify LVO patients who may otherwise be brought to the nearest hospital, which may not have the capability to perform the procedure. Accurate identification of LVO in the pre-hospital setting would allow immediate EMS transport to an MTcapable Comprehensive Stroke Center, thus reducing time-to-treatment and improving patient outcome. While various grading scales, such as the C-STAT, have been developed for this purpose, all have shown to lack sensitivity and specificity for accurate LVO determination. We hypothesize that a new scale combining common LVO presentations as positive values and those of other stroke subtypes, such as small vessel occlusion (SVO) and cardioembolic stroke (CE), as negative values will increase the accuracy of LVO determination.</p> <p><strong>Methods: </strong>This is a retrospective chart review analysis of 86 patients evaluated for stroke between January 2017-May 2018 at the Parkview Regional Medical Center with imaging confirmed LVO, SVO or CE diagnoses. &nbsp;</p> <p><strong>Results:</strong> C-STAT stroke scale had a sensitivity of 54.5% and a specificity of 86.7% in differentiating LVO from other stroke subtypes. Compared to C-STAT, our new model showed a significantly higher sensitivity of 81.8% (p=0.0038) and a nonsignificant decreased specificity of 75.0% (p=0.061).&nbsp;</p> <p><strong>Conclusion:</strong> Our findings suggest that our new scale combining common clinical presentations in LVO stroke patients as positive predictor values and those in SVO and CE stroke patients as negative predictor values may allow for a more accurate determination of LVO stroke in the pre-hospital setting without significant delay. A prospective, larger patient cohort in a pre-hospital setting is needed to validate these findings.</p> Daniel Torolira, B.S., Sara Brown, M.D., Fen-Lei Chang, M.D. Copyright (c) 2019 Daniel Torolira, B.S., Sara Brown, M.D., Fen-Lei Chang, M.D. https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23610 Wed, 09 Oct 2019 09:15:35 -0400 Identification of gene expression alterations in C. elegans adr-2 mutants using high- throughput sequencing http://journals.iupui.edu/index.php/IMPRS/article/view/23616 <p><strong>Background and Hypothesis:</strong> 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. <em>C. elegans</em> 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 <em>adr-2</em> 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.</p> <p><strong>Methods:</strong> Three replicates of neural cells isolated from wild type and <em>adr-2(-)</em> 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.</p> <p><strong>Results:</strong> I examined differential gene expression in two life stages of the WT and <em>adr-2</em> 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 <em>adr-2</em> (~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 <em>adr-2</em>.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> Jackson Townsend, Heather A. Hundley Copyright (c) 2019 Jackson Townsend, Heather A. Hundley https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23616 Wed, 09 Oct 2019 09:16:03 -0400 Development of a High-Fidelity, 3D Printed Otoplasty Surgical Simulator http://journals.iupui.edu/index.php/IMPRS/article/view/23619 <p><strong>Background and Hypothesis:</strong> 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.</p> <p>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.</p> <p><strong>Project Methods:</strong> 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.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.&nbsp;</p> Chelsey Wallace, M.S., Zahra Nourmohammadi, Ph.D., David A. Zopf, M.D., M.S. Copyright (c) 2019 Chelsey Wallace, M.S., Zahra Nourmohammadi, Ph.D., David A. Zopf, M.D., M.S. https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23619 Wed, 09 Oct 2019 09:16:35 -0400 A novel ferrochelatase inhibitor as a therapy for ocular neovascularization http://journals.iupui.edu/index.php/IMPRS/article/view/23620 <p><strong>Background and Hypothesis:</strong> 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).</p> <p>This neovascularization can be either choroidal (in the case of wet AMD) or retinal (in the case of PDR). <br> 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.</p> <p><br> 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.</p> <p><strong>Experimental Design:</strong> 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. </p> <p><strong>Results:</strong> 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.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> Sydney Waller, Sheik Pran Babu Sardar Pasha, Nathan Lambert-Cheatham, Timothy W. Corson Copyright (c) 2019 Sydney Waller, Sheik Pran Babu Sardar Pasha, Nathan Lambert-Cheatham, Timothy W. Corson https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23620 Wed, 09 Oct 2019 09:17:02 -0400 Diffusion-Weighted and Intravoxel Incoherent Motion MRI of the pregnant cervix as a tool to monitor cervical ripening http://journals.iupui.edu/index.php/IMPRS/article/view/23623 <p><strong>Background and Hypothesis:</strong> Preterm delivery is a major source of infant morbidity and mortality and is difficult to predict. The process of cervical ripening prior to delivery has known histologic changes including breakdown of collagen and increasing water content. It is hypothesized that the diffusion of water, which can be measured on MRI, will increase as the cervical water content increases. Diffusion weighted imaging (DWI) is a form of MRI that measures the random Brownian motion of water molecules (represented as the apparent diffusion coefficient (ADC)). Intravoxel incoherent motion (IVIM) MRI further subdivides the ADC into microcapillary perfusion (D*) and diffusion (D). We hypothesize that as the pregnant cervix ripens near delivery, diffusion of water within the cervix (ADC and D) will increase without changes in the capillary perfusion (D*). We sought to determine the relationship between ADC, D*, D, gestational age, and time to delivery in a cohort of volunteer pregnant females.</p> <p><strong>Experimental Design or Project Methods:</strong> DWI and IVIM MR studies from a cohort of 45 volunteer pregnant females with no known underlying fetal anomalies were examined. Subglandular and stromal cervix thickness and cervical length were measured along with the ADC, D*, and D of the subglandular and stromal cervix. Gestational history and delivery information was documented.</p> <p><strong>Results:</strong> Subglandular ADC inversely correlated with time to delivery (r=-0.393, p=0.052). Although not significant, subglandular D revealed a trend of increasing with increased gestation age (r=0.261, p=0.149). Subglandular ADC also varied inversely with thickness of the subglandular cervix (r=-0.352, p=0.047). Subglandular D* varied inversely with maternal age (r=-0.380, p=0.028).</p> <p><strong>Conclusion and Potential Impact:</strong> Our data support the hypothesized trend of increased diffusion of water within the cervix with unchanged capillary perfusion as a normal pregnancy progressed throughout the second and third trimesters. This study suggests that diffusion measures (ADC and D) follow a predictable progression during the course of a normal pregnancy and have the potential to provide a means of predicting preterm labor in the setting of preterm cervical ripening.</p> Adam Warner, BS, Gary Hutchins, PhD, Yu-Chien Wu, MD PhD, Brandon Brown, MD, Monica Forbes-Amrhein, MD PhD Copyright (c) 2019 Adam Warner, BS, Gary Hutchins, PhD, Yu-Chien Wu, MD PhD, Brandon Brown, MD, Monica Forbes-Amrhein, MD PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23623 Wed, 09 Oct 2019 09:17:24 -0400 Role of CaMKK2 in Mechanically Induced Bone Formation http://journals.iupui.edu/index.php/IMPRS/article/view/23624 <p><strong>Background and Hypothesis:</strong> 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. Ca<sup>2+</sup>/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.</p> <p><strong>Experimental Design or Project Methods:</strong> 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.</p> <p><strong>Results: </strong>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.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> Adam J. Warrick, Uma Sankar Copyright (c) 2019 Adam J. Warrick, Uma Sankar https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23624 Wed, 09 Oct 2019 09:17:52 -0400 Measuring Subjective Cognitive Decline in Older Adults: Harmonization between the Cognitive Change Index and the Measurement of Everyday Cognition Instruments http://journals.iupui.edu/index.php/IMPRS/article/view/23625 <p><strong>Background and Hypothesis:</strong> 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.</p> <p><strong>Experimental Design or Project Methods:</strong> 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, <em>APOE ε4</em> carrier status, and baseline diagnosis were also analyzed as potential relevant covariates.</p> <p><br> <strong>Results: </strong>CCI and ECog total scores were highly correlated for the self (r=0.778, p&lt;0.001) and informant (r=0.892, p&lt;0.001). The highest associations were between the mean item scores for each instrument for both self (r=0.801, p&lt;0.001) and informant (r=0.913, p&lt;0.001). Frequency distributions showed distinct patterns between self and informant total scores. Polynomial regressions for self (R<sup>2</sup>=0.673) and informant (R<sup>2</sup>=0.873) total scores were used to create a translation table between the CCI and ECog.</p> <p><strong>Conclusion and Potential Impact:</strong> 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</p> <p>[1] Rattanabannakit et al. (2016).<em> Journal of Alzheimer’s Disease</em>, 51(4), 1145-1155.</p> <p>[2] Farias et al. (2008). <em>Neuropsychology</em>, 22(4), 531-544.</p> <p>&nbsp;</p> Lindsey F. Wells, Shannon L. Risacher, Andrew J. Saykin Copyright (c) 2019 Lindsey F. Wells, Shannon L. Risacher, Andrew J. Saykin https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23625 Wed, 09 Oct 2019 09:18:24 -0400 Learning HIP: Speaking the Language of Healthcare http://journals.iupui.edu/index.php/IMPRS/article/view/23626 <p><strong>Background </strong></p> <p>The Healthy Indiana Plan serves as an insurance program, expansion of Medicaid, and unique initiative of Indiana, serving citizens near the poverty line. Elements of the program like the distinction between HIP Plus and HIP Basic, as well as the Gateway to Work program and their effects on the community have yet to be fully understood in their impact on healthcare decision making, stimulating personal responsibility, and ER utilization rates. Churches and faithbased organizations have consistently provided model forms of health support supplementation for reaching community, especially as demonstrated in the work of First Baptist Church North Indianapolis and Shepherd Community Church of the Nazarene.</p> <p><strong>Project </strong></p> <p>Methods 18 previously conducted and transcribed interviews with community leaders and community members were deductively analyzed with qualitative assessments, specifically descriptive content analysis and later framework analysis. Key themes and a framework were developed to understand and clarify responses and to produce direct recommendations.</p> <p><strong>Results</strong></p> <p>Participants in HIP Plus enjoy their insurance and experience minimal difficulty in receiving prescriptions and tests. However, remaining on HIP Plus is difficult for many participants, with the fallback of HIP Basic requiring copays that many are unable or unwilling to pay in light of greater financial priorities. Important factors arose, including: threshold knowledge to gain and maintain access, community necessitated assistance (advisory and financial), easy loss of HIP Plus status, gateway to work’s incompatibility with seasonal/temporal work, and no central administrative hub to check HIP status. Successful participants are adamant about their insurance, using phone calls, office visits, or consistent communication with navigators to ensure HIP Plus status.</p> <p><strong>Conclusion &amp; Future Directions </strong></p> <p>This study, as well as <em>Worlds Apart: Gaps</em> <em>in Life Expectancy in the Indianapolis Metro Area</em>, have contributed to a continuing study on health inequity, community perspective, and organizational activities in Indianapolis communities.</p> Matthew Wilcox, MPH, Sarah E. Wiehe, MD, MPH, Brenda L. Hudson, MA, Fiona Schicho, Ivan D. Hicks, PhD, Andrew Green, MA, David M. Craig, PhD Copyright (c) 2019 Matthew Wilcox, MPH, Sarah E. Wiehe, MD, MPH, Brenda L. Hudson, MA, Fiona Schicho, Ivan D. Hicks, PhD, Andrew Green, MA, David M. Craig, PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23626 Thu, 10 Oct 2019 10:31:26 -0400 Amyloid and Tau in Neurodegeneration http://journals.iupui.edu/index.php/IMPRS/article/view/23633 <p><strong>Background and Hypothesis:</strong><br>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.<br>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.</p> <p><br><strong>Experimental Design or Project Methods:</strong><br>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.<br>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.</p> <p><br><strong>Results:</strong><br>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.</p> <p><br><strong>Conclusion and Potential Impact:</strong><br>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.</p> Acacia Williams, Grace Hallinan, PhD, Maria Teresa Gomez Ponce, MS, Ruben Vidal, PhD Copyright (c) 2019 Acacia Williams, Grace Hallinan, PhD, Maria Teresa Gomez Ponce, MS, Ruben Vidal, PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23633 Wed, 09 Oct 2019 11:37:35 -0400 The Effect of Biofilm-Forming Bacteria on Inflammasome Activity http://journals.iupui.edu/index.php/IMPRS/article/view/23634 <p><strong>Background and Hypothesis:</strong> 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.</p> <p><strong>Methods:</strong> 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.</p> <p><strong>Results:</strong> 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.<br>The Caspase-1 activity was significantly reduced in the macrophages challenged with hyper-biofilm forming bacteria as compared to hypo-biofilm forming mutant.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> Georgia Lea Williams, BS, Suman Santra, PhD, Sashwati Roy, PhD Copyright (c) 2019 Georgia Lea Williams, BS, Suman Santra, PhD, Sashwati Roy, PhD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23634 Wed, 09 Oct 2019 11:37:49 -0400 Exposure to Adipose-Enriched Bone Marrow Increased Breast Cancer Cell Expression of Genes that Drive Tumor Metastasis http://journals.iupui.edu/index.php/IMPRS/article/view/23635 <p><strong>Background and Hypothesis:</strong> Previously thought to be inert, bone marrow adipose tissue (BMAT) has emerged as a significant fat depot that increases with age, radiation, estrogen deficiency, and high fat diet. Because bone is a preferred site for breast cancer metastasis, the lack of understanding regarding the effects of BMAT accrual on tumor cell migration and proliferation represents a significant knowledge gap in the cancer field. We hypothesized that exposure to adipokines secreted from BMAT-enriched bone marrow would induce changes in the tumor cell transcriptome.</p> <p><strong>Experimental Design or Project Methods:</strong> Twenty-week female C57Bl/6 mice were randomized and treated with control diet (5% kcal% fat) or high fat diet (HFD; 60% kcal% fat) for nine weeks. Relative to control mice, HFD mice had increased body weight, peripheral fat mass, BMAT volume, and circulating adipokines. Bone marrow was flushed and cultured for 24 hours, and control or BMAT-enriched conditioned media was collected. The adipokines leptin and MCP-1 were assayed by ELISA and found to be increased in media obtained from BMAT-enriched cultures. Human MDA-MB-231 breast cancer cells were then cultured in bone marrow conditioned media for 24 hours and tumor cells were processed for RNA sequencing.<br>Results: Exposure to soluble factors derived from BMAT-enriched cultures resulted in significant increase in the expression of genes associated with cancer cell motility and neovascularization, including SEMA4C, PLXNA1, and VEGFA.</p> <p><strong>Conclusion and Potential Impact:</strong> These data indicate that BMAT contributes to the tumor microenvironment and may drive the progression of breast cancer bone metastases.</p> Laura E. Wright, Sukanya Suresh, Jenna N. Regan, Gabriel M. Pagnotti, Sutha John, Khalid S. Mohammad, Theresa A. Guise Copyright (c) 2019 Laura E. Wright, Sukanya Suresh, Jenna N. Regan, Gabriel M. Pagnotti, Sutha John, Khalid S. Mohammad, Theresa A. Guise https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23635 Wed, 09 Oct 2019 11:38:03 -0400 Predictors of Quality of Life after Liver Transplant http://journals.iupui.edu/index.php/IMPRS/article/view/23636 <p><strong>Background and Hypothesis:</strong> 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.</p> <p><strong>Methods:</strong> 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.</p> <p><strong>Results:</strong> 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.</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> Joey Wu, Archita Desai, MD Copyright (c) 2019 Joey Wu, Archita Desai, MD https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23636 Wed, 09 Oct 2019 11:38:17 -0400 Urban-Rural Differences in Indiana Pediatric Firearm Trauma http://journals.iupui.edu/index.php/IMPRS/article/view/23637 <p><strong>Background and Hypothesis:</strong> 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.</p> <p><strong>Methods:</strong> 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&lt;0.05.</p> <p><strong>Results:</strong> 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&lt;0.05 and p&lt;0.001, respectively). Suicide frequency was higher than expected in rural counties (p&lt;0.001). Race was associated with injury intent (p&lt;0.001). Rural and midsized counties had higher than expected mortalities compared to urban counties (p&lt;0.001 and p&lt;0.05, respectively). Attempted suicides in rural areas were more likely to be associated with mortality than attempts in urban areas (p&lt;0.001).</p> <p><strong>Conclusion and Potential Impact:</strong> 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.</p> Cory Wuerch, Jodi Raymond, MPH, CSTR, CAISS, Joseph O'Neil, MD, MPF, FAAP, Matthew P. Landman, MD, MPH, FAAP, FACS Copyright (c) 2019 Cory Wuerch, Jodi Raymond, MPH, CSTR, CAISS, Joseph O'Neil, MD, MPF, FAAP, Matthew P. Landman, MD, MPH, FAAP, FACS https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23637 Wed, 09 Oct 2019 11:38:43 -0400 Designing a standard protocol for manually reviewing patient data demographics for record linkage http://journals.iupui.edu/index.php/IMPRS/article/view/23638 <p><strong>Background and Hypothesis:</strong> Accurate record linkage is essential to address fragmentation of patient data across independent healthcare organizations. To accurately evaluate record linkage methods, so-called “gold standard” data sets with labeled true matches and non-matches are needed. Human review, the process of manually assessing potentially linked patient demographic records and determining whether the record pair belongs to an idiosyncratic individual, is needed to create these datasets. However, the human review process is susceptible to bias and human error. Consequently, record linkage accuracy evaluations are prone to be biased by inaccurate gold standards. Consistent and scientifically rigorous methods for creating gold standard record linkage data sets must be developed, as none have yet been described. In this study, we describe a repeatable process for developing consistent manually reviewed datasets and analyze the results obtained from 15 human reviews of 200 record pairs following our protocol.</p> <p><strong>Experimental Design/Methods:</strong> We obtained patient records from the Indiana Network for Patient Care and Marion County Health Department. We created record pairs for manual reviews by probabilistically linking datasets using multiple blocking schemes. Two-hundred record pairs were then manually reviewed by 15 different individuals and the results were analyzed.</p> <p><strong>Results:</strong> Across the 200 record pairs reviewed by 15 reviewers, 155 were nondiscordant pairs whereas 45 were discordant, 40 among which were the result of outliers.</p> <p><strong>Conclusion and Potential Impact:</strong> From the record pair evaluation results, some empirical rules can be established for the process of manual review, though the nuances of evaluation reasoning require more discussion and a larger sample size. Nonetheless, establishing a standard for manual reviewing is a step towards better health care and complete patient records.</p> Sen Xiong, Shuan Grannis, MD, MS, FAAP Copyright (c) 2019 Sen Xiong, Shuan Grannis, MD, MS, FAAP https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23638 Wed, 09 Oct 2019 11:38:57 -0400 Physiological Responses to Hypoxia in the Absence of Brain Glycogen http://journals.iupui.edu/index.php/IMPRS/article/view/23639 <p><strong>Background and Hypothesis:</strong> Glycogen is a highly branched polymer of glucose and is an important form of energy storage in mammals. The brain is able to form glycogen in astrocytes and neurons via glycogen synthase and branching enzyme. Once formed, brain glycogen functions as the only stored energy source for these cells. Various physiological roles for brain glycogen have been hypothesized, including memory consolidation and sleep regulation, as well as a protective role during various physiological stressors, such as hypoglycemia and hypoxia. For instance, rat brain glycogen levels were decreased 10 minutes after vaginal birth, but not after a C-section. This suggested that cerebral hypoxia experienced during vaginal birth induced the utilization of brain glycogen to minimize neurodegeneration during the hypoxic event. Symptoms of hypoxia can range from tachycardia, tachypnea, shortness of breath, and diaphoresis to confusion, loss of motor coordination and cognitive function, neurodegeneration, and brain death. The many causes of hypoxia include lungs diseases (COPD, pneumonia, pulmonary edema), CNS depressants (opiates), heart problems (CHF), anemia, and obstructive sleep apnea (OSA). We hypothesized that the lack of brain glycogen would cause a noticeable detrimental effect to the survival time and physiologic functions of mice exposed to acute hypoxia.</p> <p><strong>Experimental Design or Project Methods</strong>: We subjected mice, with or without glycogen synthase disrupted in the brain, to carbon dioxide- or nitrogen-induced hypoxia and monitored effects on brain glycogen levels, behavior, and survival time.</p> <p><strong>Results:</strong> We found that mice lacking brain glycogen exhibited the characteristic physiologic responses to hypoxia, but expired ~50% sooner than mice with brain glycogen.</p> <p><strong>Conclusion and Potential Impact:</strong> These results provide evidence that brain glycogen is imperative in responding to hypoxic events. Further, these findings suggest that brain glycogen may protect patients with OSA against other comorbidities, especially neurodegeneration and cognitive impairment.</p> Taylor Zike, Justin J. Crowder, Bartholomew A. Pederson Copyright (c) 2019 Taylor Zike, Justin J. Crowder, Bartholomew A. Pederson https://creativecommons.org/licenses/by/4.0 http://journals.iupui.edu/index.php/IMPRS/article/view/23639 Wed, 09 Oct 2019 11:39:12 -0400